Patent Application
20230381117
The invention in general relates to compositions for weight management. Specifically, the invention relates to compositions containing bisdemethoxycurcumin for the management of obesity and hyperlipidemia.
Obesity is considered to be the leading health risk for the development of various disorders like hypertension, type 2 diabetes, heart disease, stroke, osteoarthritis, and mental illness. Globally, more than 1 in 10 individuals are obese and about 36% of American adults are obese. Obesity results due to imbalance between the energy content of food eaten and energy expended by the body to maintain life and to perform physical work. Such an energy balance framework is a potentially powerful tool for investigating the regulation of body weight.
Conversion of white adipose tissue to brown or beige/brite is reported as an effective mechanism to utilize the undue energy abundance and increasing the energy expenditure. This “energy balance” wherein there is a balance between the calories consumed and expended, is vital for any weight loss regimen. The conversion to BAT accelerates the expenditure of the stored “energy” and brings about a balance in energy homeostasis. The role of brown adipose tissue (BAT) is well described in the following prior arts:
Drugs and/or natural molecules that facilitate the conversion of white to brown adipocytes are effective in the treatment/management of obesity related conditions. However, we need a better understanding of the components involved in energy expenditure and their interactions over various time scales to explain the natural history of conditions such as obesity and to estimate the magnitude and potential success of therapeutic interventions. (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 April; 95(4): 989-994).
There exists a vast knowledge about natural plant based ingredients for the management of obesity and related disorders. It is well known in the scientific art regarding the compounds obtained from Curcuma species, specifically curcuminoids, and their role in therapeutic management of various diseases and disorders are well documented. The metabolites of curcumin—demethoxycurcumin, bisdemethoxycurcumin, tetrahydrocurcuminoids, hexahydrocurcuminoids, and octahydrocurcuminoids are garnering much attention owing to their similar and superior efficacy over curcumin (Majeed et al., Reductive Metabolites of Curcuminoids, Nutriscience Publishers LLC, 2019). Bisdemethoxycurcumin is reported to have superior effect over curcumin in managing certain pathological conditions (Franco Caveleri, U.S. Pat. No. 10,945,970). Lai et al., (Bisdemethoxycurcumin Inhibits Adipogenesis in 3T3-L1 Preadipocytes and Suppresses Obesity in High-Fat Diet-Fed C57BL/6 Mice, J Agric Food Chem. 2016 Feb. 3;64(4):821-30. doi: 10.1021/acs.jafc.5b05577) also reports the anti-obesity of bisdemethoxycurcumin. However, there exists an unmet industrial need for a curcuminoid composition that is effective in, bringing about energy balance and weight loss. The present invention discloses a curcuminoid composition, enriched in bisdemethoxycurcumin for the management of obesity and related conditions.
The principle objective of the invention is to disclose the anti-obesity effect of a composition comprising not less than 20% w/w bisdemethoxycurcumin by bringing about weight loss and energy balance.
It is another objective of invention to disclose the hypolipidemic effects of ability a composition comprising not less than 20% w/w bisdemethoxycurcumin.
The present invention fulfils the above such objectives and provides further related advantages.
In a most preferred embodiment, the present invention discloses a method for therapeutic management of obesity in mammals, said method comprising steps of identifying a mammal with obesity and administering effective concentration of a composition comprising not less than 20% w/w bisdemethoxycurcumin to said mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-50% w/w curcumin.
In another most preferred embodiment, the present invention discloses a composition comprising not less than 20% w/w bisdemethoxycurcumin for therapeutic management of obesity in mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-50% w/w curcumin.
In another preferred embodiment, the invention discloses a method of therapeutic manauement of hyperlipidemia in mammals, said method comprising step of a) identifying a mammal with hyperlipidemia and b) administering an effective concentration of a composition comprising not less than 20% w/w bisdemethoxycurcumin to said mammals to bring about the effects of (i) reducing the amount of total blood cholesterol levels; (ii) reducing the concentrations of low density lipoproteins (LDL) and very low density lipoproteins (VLDL); (iii) increasing the concentrations of high density lipoproteins (HDL) and (iv) reducing concentrations of serum triglycerides, in the blood of said mammals.
In another preferred embodiment, the invention discloses a composition comprising not less than 20% w/w bisdemethoxycurcumin for the therapeutic management of hyperlipidemia in mammals.
Other features and advantages of the present invention will become apparent from the following more detailed description, which illustrate, by way of example, the principle of the invention.
In a most preferred embodiment, the present invention discloses a method for therapeutic management of obesity in mammals, said method comprising steps of identifying a mammal with obesity and administering effective concentration of a composition comprising not less than 20% w/w bisdemethoxycurcumin to said mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-50% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 25-45% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 30-50% w/w curcumin. In yet another related aspect, the total curcuminoids in the composition are in the range of 20-95% w/w. In a related aspect, the effective concentration of the composition is 50 mg/kg bodyweight. In a related aspect, management of obesity is brought about by inhibition of adipogenesis, decreasing body weight and organ weight, increasing protein content, decreasing elevated levels of liver enzymes, decreasing circulating glucose and lipid levels, inhibiting lipid deposition and enhancing energy expenditure. In another related aspect, energy expenditure is enhanced by (a) increasing the expression of genes and proteins that function individually or in combination to specifically recruit brown adipocytes or beige or brite adipocytes, (b) inducing brown or beige or brite like phenotype in white adipocyte depots, (c) inducing enhanced expression of mitochondrial uncoupling protein-1 (UCP-1) to result in increased mitochondrial thermogenesis in differentiated brown adipocytes and beige or brite adipocytes. In a related embodiment, the genes and proteins are at least one factor selected from the group consisting of comprising PRDM16, PGC-1α and BMP7. In another related aspect, the liver enzymes are selected from the group consisting of aspartate transaminase and alanine transaminase. In another related aspect, the mammal is human. In a related embodiment, the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.
In another most preferred embodiment, the present invention discloses a composition comprising not less than 20% w/w bisdemethoxycurcumin for therapeutic management of obesity in mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-50% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 25-45% wlw curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 30-50% w/w curcumin. In yet another related aspect, the total curcuminoids in the composition are in the range of 20-95% w/w. In a related aspect, the effective concentration of the composition is 50 mg/kg bodyweight. In a related aspect, management of obesity is brought about by inhibition of adipogenesis, decreasing body weight and organ weight, increasing protein content, decreasing elevated levels of liver enzymes, decreasing circulating glucose and lipid levels, inhibiting lipid deposition and enhancing energy expenditure. In another related aspect, energy expenditure is enhanced by (a) increasing the expression of genes and proteins that function individually or in combination to specifically recruit brown adipocytes or beige or brite adipocytes, (b) inducing brown or beige or brite like phenotype in white adipocyte depots, (c) inducing enhanced expression of mitochondrial uncoupling protein-1 (UCP-1) to result in increased mitochondrial thermogenesis in differentiated brown adipocytes and beige or brite adipocytes. In a related embodiment, the genes and proteins are at least one factor selected from the group consisting of comprising PRDM16, PGC-1α and BMP7. In another related aspect, the liver enzymes are selected from the group consisting of aspartate transaminase and alanine transaminase. In another related aspect, the mammal is human. In a related embodiment, the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.
In another preferred embodiment, the invention discloses a method of therapeutic management of hyperlipidemia in mammals, said method comprising step of a) identifying a mammal with hyperlipidemia and b) administering an effective concentration of a composition comprising not less than 20% w/w bisdemethoxycurcumin to said mammals to bring about the effects of (i) reducing the amount of total blood cholesterol levels; (ii) reducing the concentrations of low density lipoproteins (LDL) and very low density lipoproteins (VLDL); (iii) increasing the concentrations of high density lipoproteins (HDL) and (iv) reducing concentrations of serum triglycerides, in the blood of said mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-50% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 25-45% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 30-50% w/w curcumin. In yet another related aspect, the total curcuminoids in the composition are in the range of 20-95% w/w. In a related aspect, the effective concentration of the composition is 50 mg/kg bodyweight. In a related embodiment, the medical cause of hyperlipidemia is high fat diet induced obesity. In another related aspect, the mammal is human. In a related embodiment, the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.
In another preferred embodiment, the invention discloses a composition comprising not less than 20% w/w bisdemethoxycurcumin for the therapeutic management of hyperlipidemia in mammals. In a related aspect, therapeutic management if brought about by (i) reducing the amount of total blood cholesterol levels; (ii) reducing the concentrations of low density lipoproteins (LDL) and very low density lipoproteins (VLDL); (iii) increasing the concentrations of high density lipoproteins (HDL) and (iv) reducing concentrations of serum triglycerides, in the blood of said mammals. In a related embodiment, the composition further comprises of 10-35% w/w demethoxycurcumin and 10-15% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 25-45% w/w curcumin. In another related aspect, the composition comprises of 20-50% w/w bisdemethoxycurcumin, 10-25% w/w demethoxycurcumin and 30-50% w/w curcumin. In yet another related aspect, the total curcuminoids in the composition are in the range of 20-95% w/w. In a related aspect, the effective concentration of the composition is 50 mg/kg bodyweight. In a related embodiment, the medical cause of hyperlipidemia is high fat diet induced obesity. In another related aspect, the mammal is human. In a related embodiment, the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.
Specific illustrative examples enunciating the most preferred embodiments are included herein below
Methodology
Preparation of Stock Solutions
The composition comprising not less than 20% w/w bisdemethoxycurcumin, w/w demethoxycurcumin and 10-50% w/w curcumin was isolated from Curcuma longa using a process known in the scientific art. The composition is also available commercially from Sami-Sabinsa Group as Curcumin BD3 Complex™ (also known as AC3 complex). The specific range of the composition used in the experiment include 32% w/w bisdemethoxycurcumin, 25% demethoxycurcumin and 38% curcumin (total curcuminoids content: 73%). It is to be noted that the range tested is merely illustrative and the results are application to the aforementioned ranges of the composition.
The stock of different test ingredients—AC3 complex (AC3), Curcumin C3 complex (composition enriched in curcumin (95% curcuminoids) which comprises 75-81% curcumin, 15-19% demethoxycurcumin, and 2.2-6.5% bisdemethoxycurcumin—C3 complex), demethoxycurcumin (DMC), bisdemethoxycurcumin (BDMC) and curcumin (95%) of 10 mg/ml was prepared in DMSO and filtered through 0.2 micron syringe filter. Stock was diluted 1000 times in DMEM to get 10 μg/ml final concentration and serially diluted. Insulin (Hi Media) was bought as a solution at a concentration of 20 mg/ml. This was diluted to 1 μg/ml in DMEM. IBMX (Sigma)—Stock of 5 mM was prepared in DMEM, and diluted 10 times to be used at a final concentration of 0.5 mM. Dexamethasone (Sigma)—A stock of 10 μM was prepared in DMEM and diluted 40 times to get a final concentration of 0.25 μM.
Cell Culture
Mouse 3T3-L1 pre-adipocytes were cultured in DMEM media containing 25 mM glucose, 10% heat-inactivated new born calf serum (NBCS) and antibiotics at 37° C. and 5% CO2. When the cells were 70-80% confluent, they were trypsinized, washed and seeded in 48 well plates at a density of 3×104 cells per well. Cells were induced to differentiate 2 d after reaching confluence (day 0), by supplementing DMEM media containing 10% Fetal Bovine Serum (FBS) along with 1μg/mL insulin, 0.25 μM dexamethasone, 0.5 mM 1-methyl-3-isobutyl-xanthine (IBMX) and different concentrations of test ingredients. After differentiation for two days, The media was replaced with DMEM containing insulin (1 μg/mL) and different concentrations of test ingredients on alternate days until the cells were processed after 3rd progression. Untreated cells grown in differentiation media and undifferentiated cells grown in NBCS media were taken as adipogenesis positive and negative controls for the experiment. Quantification of triglycerides accumulated in adipocytes was done by Oil red O staining.
RNA Extraction
Cells were seeded in 6 well plates at a density of 2×105 cells per well, treatment was done as mentioned above, harvested after second progression on day 7 and total RNA was extracted using the Trizol method. Extracted RNA was treated with DNAse I to remove any contaminating DNA and again extracted using phenol: chloroform: isoamyl alcohol extraction (24:25:1). Quality of RNA was determined by checking the absorbance at 260/280 nm using a Nanodrop (Thermo)
Quantitative Real Time PCR
2 μg of total RNA was taken for cDNA synthesis using Revert-aid First Strand cDNA synthesis kit (Thermo Scientific).). Quantitative RT-PCR analysis was performed to determine the expression of brown fat specific genes in Roche Light cycler 96 using SYBR Green master mix (Thermo Scientific). β actin was used as a house keeping gene. The relative RNA abundance of BAT genes was normalized to the housekeeping β actin gene and expressed as delta delta CT (equivalent to fold change transformed by Log2).
Primer sequence: The primers used for determining the expression of brown fat specific genes and genes related to adipogenesis is given in table 1
Results
The lipids accumulated in adipocytes were quantified by Oil red O staining. Bisdemethoxycurcmin (BDMC) showed a improved reduction in lipid accumulation in adipocytes (
The AC3 complex also significantly increased the brown adipose tissue specific genes. The expression of UCP1, PRDM16, PGC1α and BMP7 was increased (
Methods
Animals—C57/BL6 mice, 6-8 weeks of age and 8 animals/Group (4 Male and 4 Female) were used for the study. Animals were housed under standard laboratory conditions, air-conditioned with adequate fresh air supply (12-15 Air changes per hour), room temperature 20.2-23.5° C. and relative humidity 58-64% with 12 hours fluorescent light and 12 hours dark cycle. The temperature and relative humidity was recorded once daily.
Feed: The animals were fed with Normal diet (9 kcal/day) and High fat diet (50 kcal/day) throughout the acclimatization and experimental period.
Water was provided along with High Fat Diet to the animals throughout the acclimatization and experimental period. Water from water filter cum purifier was provided in animal feeding bottle with stainless steel sipper tubes.
All the studies were conducted according to the ethical guidelines of CPCSEA after obtaining necessary clearance from the committee (Approval No: 790/03/ac/CPCSEA).
The design of the study groups is depicted in table 4.
Body weight of the animals was recorded in all the days of experimental period. At the end of the experimental period, the animals were sacrificed by cervical dislocation. Blood was collected and Serum was separated by centrifugation and used for the analysis of biochemical parameters. The organs such as Liver Kidney, Spleen and Pancreas and Fat Tissues (Retroperitoneal, Peri-gonadal and Mesenteric) were dissected out and washed in phosphate buffered saline.
Efficacy Measurement
The following parameters were measured in the above groups:
Results
Body Weight
The results indicated that BDMC inhibited weight gain in the animals fed with high fat diet (
Liver Weights and Liver Enzymes
The administration of different compositions did not adversely affect the weight of the liver, suggesting that BDMC does not induce any adverse effects in critical orizans, however slight decrease in the liver weight by BDMC was observed indicating that BDMC decreases fat content in the liver (
Lipid profile: The high fat diet increased the levels of total cholesterol, LDL, VLDL and triglycerides in the serum of study animals. High fat diet, co administered with BDMC, significantly reduced the total cholesterol (
BDMC and AC3 treatment showed inhibition of adipogenesis in vitro and induced the conversion of white adipose tissue to brown or brite/beige thereby increasing fat utilisation and energy metabolism. The in vivo results indicated that both BDMC and Ac3 complex were effective in significantly reducing body weight and visceral fat accumulation. Over all, BDMC induces weight loss, reduces viceral fat and maintains health of key organs. BDMC can be administered along with other plant ingredients isolated from the different medicinal plants from the group of, but not limited to, Nigella sativa, Oroxylum indicum, Garcinia cambogia, Garcinia indica, Pterocarpus marsupium, Cyperus rotundus, Coleus forskohlii, Zingeber cassumunar, Withania somnifera, Polygonum cuspidatum, Boswellia serrata, Artocarpus sp., Emblica officinalis, Curcuma longa, Terminalia arjuna, specifically with the ingredients which include, but not limited to, Calebin, Oroxylin A, withanolides, thymoquinone, thymohydroquinione, scirpusin A, scirpusin B, piceatannol, pterocarposide, forskolin, glucogallin, resveratrol, orxyresveratrol, boswellic acids, arjunolic acid, arjunoglucosides, garcinol. The BDMC composition can also be added to different probiotic formulations for effective weight loss regime.
The composition is formulated along with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents, stabilizing agents, dispersible gums, bioavailability enhancers or carriers and administered orally in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies or eatables.
In a related aspect the bioavailability enhancer is selected from the group of piperine (BioPerine®), quercetin, garlic extract, ginger extract, and naringin. In another related aspect, the stabilizing agent is selected from the group consisting rosmarinic acid, butylated hydroxyanisole, butylated hydroxytoluene, sodium metabisulfite, propyl gallate, cysteine, ascorbic acid and tocopherols. In yet another related aspect, the dispersible gums are selected from the group consisting of Agar, Alginate, Carrageenan, Gum Arabic, Guar Gum, Locust Bean Gum, Konjac Gum, Xanthan Gum and Pectin.
Tables 5-9 provide illustrative examples of nutraceutical formulations containing bisdemethoxycurcumin
The above formulations are merely illustrative examples, any formulation containing the above active inuredient intended for the said purpose will be considered equivalent.
Other modifications and variations of the invention will be apparent to those skilled in the art from the foregoing disclosure and teachings. Thus, while only certain embodiments of the invention have been specifically described herein, it will be apparent that numerous modifications may be made thereto without departing from the spirit and scope of the invention and is to be interpreted only in conjunction with the appended claims.
This is a U.S. national phase application of PCT/US21/63691 filed on 16 Dec. 2021, claiming priority to U.S. provisional application No. 63126920 filed on 17 Dec. 2020, the contents of which are incorporated herein by reference.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US21/63691 | 12/16/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63126920 | Dec 2020 | US |
