CURCUMINOID COMPOSITIONS

Abstract

The present invention relates to compositions comprising curcuminoids; in particular, compositions comprising curcuminoids that are highly water soluble. The present invention also relates to processes for providing such compositions and uses of such compositions.

Claims

1. A composition comprising: (i) at least about 20% curcuminoids by weight of the composition; (ii) gum arabic; and (iii) an extract obtained or obtainable from quillaja, wherein the composition comprises particles having an average diameter of from about 100 nm to about 10000 nm.

2. A composition according to claim 1, wherein the particles have an average diameter of from about 100 nm to about 700 nm

3. A composition according to claim 1 or 2 further comprising plant and/or vegetable oil.

4. A composition according to any one of claims 1 to 3, wherein the quillaja extract is present in an amount from about 0.1 to about 5% by weight of the composition.

5. A composition according to any one of the preceding claims, wherein the gum arabic is present in an amount from about 40 to about 60% by weight of the composition.

6. A composition according to any one of the preceding claims, wherein the quillaja extract comprises at least 50% saponins.

7. A composition according to any one of the preceding claims, wherein the quillaja extract is in the form of a solid.

8. The use of a composition according to any one of claims 1 to 7 as nutraceutical formulation, a dietary or food product for humans or animals (such as functional food formulations, i.e. food, drink, feed or pet food or a food, drink, feed or pet food supplements), a nutritional supplement, a fragrance or flavouring, a pharmaceutical or veterinary formulation, an oenological or cosmetic formulation.

9. The use of a composition according to any one of claims 1 to 7 in the preparation of a nutraceutical formulation, a dietary or food product for humans or animals (such as functional food formulations, i.e. food, drink, feed or pet food or a food, drink, feed or pet food supplements), a nutritional supplement, a fragrance or flavouring, a pharmaceutical or veterinary formulation, an oenological or cosmetic formulation.

10. A nutraceutical formulation, a dietary or food product for humans or animals (such as functional food formulations, i.e. food, drink, feed or pet food or a food, drink, feed or pet food supplements), a nutritional supplement, a fragrance or flavouring, a pharmaceutical or veterinary formulation, an oenological or cosmetic formulation consisting of, consisting essentially of or comprising a composition as defined in any one of claims 1 to 7.

11. The use according to claims 8 or 9, wherein the nutraceutical formulation, dietary or food product for humans or animals (such as functional food formulations, i.e. food, drink, feed or pet food or a food, drink, feed or pet food supplements), nutritional supplement, fragrance or flavouring, pharmaceutical or veterinary formulation, oenological formulation or cosmetic formulation further comprises pharmaceutically/veterinary ingredients, such as excipients or carriers or (function) food acceptable ingredients and mixtures thereof as appropriate.

12. The nutraceutical formulation, dietary or food product for humans or animals (such as functional food formulations, i.e. food, drink, feed or pet food or a food, drink, feed or pet food supplements), nutritional supplement, fragrance or flavouring, pharmaceutical or veterinary formulation, oenological formulation or cosmetic formulation according to claim 9, further comprising pharmaceutically/veterinary ingredients, such as excipients or carriers or (function) food acceptable ingredients and mixtures thereof as appropriate.

13. A process for the preparation of a composition as defined in claims 1 to 7, wherein the process comprises the steps of: (i) preparing an aqueous solution of curcuminoids; (ii) mixing the aqueous solution from (i) with an aqueous gum arabic solution and extract obtained or obtainable from quillaja and optionally a plant and/or vegetable oil to provide an emulsion; and optionally (iii) drying the product of (ii) to provide a composition comprises particles having an average diameter of from about 100 nm to about 10000 nm.

Description

BRIEF DESCRIPTION OF FIGURES

[0151] FIG. 1—Chemical structures of the curcuminoids from turmeric.

[0152] FIG. 2—Phase I and phase II metabolites of curcumin

[0153] FIG. 3—Bacterial metabolites of curcumin

[0154] FIG. 4—the effect of pH on the composition of the invention colour dissolved in demineralized water (0.4%).

[0155] FIG. 5—the DLS profile of loaded of composition of the invention.

[0156] FIG. 6—The Z-potential of the composition of the invention and individual components of the composition in different pHs.

[0157] FIG. 7—Scanning electron microscopy (SEM) image of the composition of the invention at ×300.

[0158] FIG. 8—Scanning electron microscopy (SEM) image of the composition of the invention at ×1300.

[0159] FIG. 9—Scanning electron microscopy (SEM) image of the composition of the invention at ×9200.

[0160] FIG. 10—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the standard curcumin extract. Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0161] FIG. 11—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the standard curcumin extract with data normalized to the TB+colon 48 h sample (100%). Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0162] FIG. 12—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the turmeric phytosome formulation. Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0163] FIG. 13—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the turmeric phytosome formulation with data normalized to the TB+colon 48 h sample (100%). Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0164] FIG. 14—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the quillaja based formulation. Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0165] FIG. 15—LDH activity measured on the apical supernatant collected from the Caco-2 cells treated with samples coming from the different gastrointestinal tract compartments after 4 h of incubation, data from the quillaja based formulation with data normalized to the TB+colon 48 h sample (100%). Bars depict Mean±SEM. PP: pure product; SI: small intestine; ST: stomach; TB: transport buffer. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively, compared to TB+colon 48 h.

[0166] FIG. 16—Evolution of the food intake over the acclimatization and habituation periods of groups of mice sacrificed at the same time point. Mean daily food intake (g) of the different experimental groups during the acclimatization (J0-J7) and habituation (J8-J14) periods.

[0167] FIG. 17—Evolution of body weight over the acclimatization and habituation periods of groups of mice sacrificed at the same time point. Mean body weight (g) of the different experimental groups during the same periods. Data are represented as mean±SEM.

[0168] FIG. 18—Time-course of total curcuminoids levels (sum of curcumin, DMC, BDMC and their relative metabolites curcumin glucuronide and sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) in mouse plasma after a single oral dose of Turmeric Phytosome formulation versus standard turmeric extract containing 300 mg/kg of curcuminoids. *, **, ***: The Turmeric Phytosome formulation significantly different (post-hoc t-test) at each time point from standard turmeric extract (p<0.05, p<0.01, p<0.001 respectively).

[0169] FIG. 19—Time-course of total curcuminoids levels (sum of curcumin, DMC, BDMC and their relative metabolites curcumin glucuronide and sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) in mouse plasma after a single oral dose of the composition as used in the methods/uses of the invention versus standard turmeric extract containing 300 mg/kg of curcuminoids. *, **, ***: The composition as used in the methods/uses of the invention significantly different (post-hoc t-test) at each time point from standard turmeric extract (p<0.05, p<0.01, p<0.001 respectively).

[0170] FIG. 20: Total curcuminoids and metabolites (sum of curcumin, DMC, BDMC and their relative metabolites curcumin glucuronide and sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) concentration (ppm) as a function of time (h) after consumption of the different formulations (n=72 per formulation). .sup.□, .sup.□□: Turmeric Phytosome significantly different from standard turmeric extract (p<0.05, p<0.01 respectively); *, **, ***: The composition as used in the methods/uses of the invention significantly different from standard turmeric extract (p<0.05, p<0.01, p<0.001 respectively).

[0171] FIG. 21—Corresponding area under the curve AUC(0-8 h) of the different formulations.

[0172] FIG. 22—Corresponding area under the curve AUC(0-∞) of the different formulations.

[0173] FIG. 23—Cmax (concentration at Tmax) of the different formulations.

[0174] FIG. 24—Graphical representation of Dose-normalized of AUC.sub.0-24h for the ITT population.

[0175] FIG. 25—Graphical representation of Dose-normalized of AUC.sub.0-8h for the ITT population.

[0176] FIG. 26—Graphical representation of Dose-normalized of AUC.sub.0-∞ for the ITT population.

[0177] FIG. 27—Graphical representation of AUC.sub.0-24 h for the ITT population.

[0178] FIG. 28—Graphical representation of AUC.sub.0-8h for the ITT population.

[0179] FIG. 29—Graphical representation of AUC.sub.0-∞ for the ITT population.

[0180] FIG. 30—Graphical representation of the normalised Cmax for the ITT population.

[0181] FIG. 31—Graphical representation of the Cmax for the ITT population.

[0182] FIG. 32—Graphical representation of the relative bioavailability between 0 and 24 hours for the ITT population.

[0183] FIG. 33—Graphical representation of the relative bioavailability between 0 and 8 hours for the ITT population.

[0184] FIG. 34—Graphical representation of the relative bioavailability between 0 and infinity for the ITT population.

[0185] FIG. 35—Graphical representation of the half-life for the ITT population.

[0186] FIG. 36—Graphical representation of terminal elimination rate constant for the ITT population.

[0187] FIG. 37—Graphical representation of the Tmax for the ITT population.

EXAMPLES

[0188] The present invention will be further described by reference to the following, non-limiting examples.

Example 1—Preparation of Alkaline Organic Curcumin Solution

[0189] A mixture of curcuminoids in water was prepared using an organic purified curcuminoid extract (at least at 10% but preferably at 95% of purity (total curcuminoids)) in distilled water (3 volumes powder weight/water).

Example 2—Preparation of a Composition of the Invention

[0190] A 58% gum Arabic mixture (substrate) was prepared using distilled water (3 volumes powder weight/water). 500 ml of the aqueous gum arabic solution was added 500 ml of the curcuminoid solution prepared in Example 1 under agitation (5000 rpm) and to this was added 5% of organic sunflower oil and 2% of organic quillaja standardized in saponins.

[0191] The resulting mixture was agitated at 5000 rpm for ten minutes. The resulting emulsion was then spray dried.

[0192] Example 3—Characterization of a Composition of the Invention The size and morphology of the composition of the invention was analyzed by dynamic light scattering (DLS), and zeta potential (Z-potential), and scanning electron microscopy (SEM). For DLS and zeta-potential analyses a Zetasizer Nano ZS (NanoZS90, Malvern Instrument Ltd., UK) with a He/Ne laser (λ=633 nm) at a fixed scattering angle of 90° at temperature of (25±0.1° C.) was used.

[0193] The samples used were in liquid emulsion form (last step before drying). The samples were suspended in demineralized water at a volume concentration of 0.4% and 1 minute of ultrasound was applied. DLS analysis was immediately performed at these samples (measure time=60 seconds). The analysis of zeta-potential was performed in a large pH range (from 2 to 11).

[0194] The samples were prepared and analyzed at different pHs as follows by the use of 0.1M HCl and 0.1M NaOH solutions. The 10 samples (pH=2, 3, 4, 5, 6, 7, 8, 9, 10 and 11) obtained were stored at room temperature (23° C.) (FIG. 4).

[0195] As shown in FIG. 4, the colour of the composition of the invention in water is driven by pH. The keto form (yellow) is the predominant form present in solution when pH range vary from acid to neutral (from 2- to 7). At pH 8 and 9 colour solution turns to orange, and at pH 10 and 11 a translucid reddish colour is predominant. The colour change is due to sequential deprotonation of hydroxyl-groups of curcumin molecule driven by pH increasing which gives a higher solubility and instability to curcumin.

[0196] The DLS analysis results are showed in FIG. 5. There are two groups of particle size individuals. One centralized at 616±160 nm (20.8% of total individuals) and the most interesting one centralized at 188±42 nm (79.2% of the total individuals).

[0197] The mean hydrodynamic particle size of loaded curcumin in water solution (pH 5.4) was found to be 476.5 nm with a PDI (polydispersity index) of 0.337.

[0198] FIG. 6 shows the Z-potential of the composition of the invention at different pHs (from 2 to 11). The higher Z-potential, the more unstable mixture is. The composition of the invention has a negative Z-potential between pH 2 and pH 11. The particles are negatively charged in the aqueous phase. At pH 2 the Z-potential is close to 0 (isoelectric point: pH for which the potential is zero), where we have an instability zone of the emulsions.

[0199] Between pH 2 and pH 4, the Z-potential is relatively low (<25 mV) and at pH greater than or equal to 4, the sample enters a zone of stability. This stability is strongly confirmed from pH 5. A rapid shift in the isoelectric point was observed in pH 8.0 the zeta-potential of the loaded curcumin was surprisingly higher in pH range from 2 to 7. In the aqueous phase at pHs<4.0, loaded curcumin are usually at their lowest surface energy state. At pH 8.0, the loaded curcumin is likely to orient its low electronic charge side towards Arabic gum and expose its high electronic charge side to interact with water, which leads to an elevated zeta-potential.

[0200] FIG. 6 clearly shows that the claimed composition is stable when dispersed in an aqueous solution at pH greater than 4.

Example 4—Particle Size Distribution (PSD) of a Composition of the Invention Using CQ-MO-304

[0201] Materials and Reagents

[0202] Material—Mastersizer 3000 from Malvern Instrument, or equivalent; [0203] Hydro 2000SM sample dispersion unit, or equivalent (for liquid phase), [0204] Malvern AERO S sample dispersion unit, or equivalent (for solid phases).

[0205] Reagent—Water

[0206] Procedure [0207] Analytical parameters [0208] background time: 10 seconds [0209] Measurement time@10 seconds [0210] Refractive index of distilled water: 1.33 [0211] Result calculation: general purpose [0212] Pump/stir speed: 1800 RPM [0213] liquid dispersant: water [0214] solid dispersant: ambient air [0215] Specific parameters [0216] 100705 (refractive index: 1, adsorption: 1) [0217] 100019 (refractive index: 1, adsorption: 2) [0218] 3CAA0075 and 3CAA0076 (Composition of the invention)

[0219] A sample of the composition of the invention was mixed with distilled water and a sample was tested either using a Hydro 2000SM unit or Mastersizer 3000 (using a Scirocco 2000 unit).

[0220] Results

[0221] Several batches of the composition of the invention, obtained after drying and milling, were tested according to the above-mentioned method. The results are provided in Table 1 below.

TABLE-US-00001 TABLE 1 Particle Size Distribution of the composition of the invention (where (D90) corresponds to 90% of the particle size population, and (D4:3) corresponds to the volume moment mean of the particle size population). PS (D90) PS (D4:3) Ball Mill Reference Product name in μm in μm 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.3 1.7 3COA0004 Organic Turmipure Enhanced 30% - 3.4 1.6 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.5 2.1 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 2.6 1.6 Ball Milled 3CAA0076 Turmipure Enhanced 30% - Ball Milled 2.4 1.4 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.5 1.8 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.2 1.5 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.1 1.5 3CAA0076 Turmipure Enhanced 30% - Ball Milled 2.7 1.3 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.1 1.5 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.5 1.7 3CAA0076 Turmipure Enhanced 30% - Ball Milled 2.7 1.4 3CAA0076 Turmipure Enhanced 30% - Ball Milled 4.4 2.0 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.1 1.5 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.4 1.6 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.2 1.6 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.9 1.7 3CAA0076 Turmipure Enhanced 30% - Ball Milled 3.9 1.8 3COA0004 Organic Turmipure Enhanced 30% - 2.9 1.4 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.1 1.4 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.1 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.3 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.4 1.6 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.6 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.1 1.4 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 2.9 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.1 1.7 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 2.9 1.4 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 2.96 1.41 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.39 1.59 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3.2 1.5 Ball Milled 3COA0004 Organic Turmipure Enhanced 30% - 3 1.4 Ball Milled

Example 5—Morphology of a Composition of the Invention (by Scanning Electron Microscopy, SEM)

[0222] For SEM analysis samples were prepared as follow: The composition of the invention in powder form was deposited on the sample holder by simply dusting. It was followed by metallization of platinum/palladium deposits and then observation and shooting by scanning electron microscope equipped with X-ray detector in energy dispersion.

[0223] The SEM images shown in FIGS. 7, 8 and 9 provide the visualization of the composition of the invention.

[0224] The composition of the invention shows conjugate self-assembled to spherical micelles with size of +/−170 nm. The roughly spherical morphology in SEM analysis corroborated the size measurement analysis done by dynamic light scattering technique.

[0225] From SEM, it can be seen that the particles in the composition of the invention have clearly shown outer coat of chitosan which was absent in uncoated lecithin nanoparticles. Curcumin was found to be well dispersed in the lecithin core of the nanoparticles. SEM measurements also corroborated evidence of roughly spherical geometry and the surface roughness indicate surface absorption. It suggests that the driving force of this type of adsorption is either direct electrostatic interaction or ion-ion interaction.

Example 6—Testing the Effect of a Composition of the Invention to Enhance the Bioavailability of Curcuminoids in an In-Vivo Model of the Human Gastrointestinal Tract and Intestinal Absorptive Cells

[0226] The human gastrointestinal tract (GIT) is one of the major entry gates to the human body. Upon oral ingestion of foods, drinks or pharmaceuticals, the gut is the first site of contact between the ingested products and the host. In order to exert their biological activity, compounds have first to pass the stomach where the acid environment and the presence of digestive enzymes may lead to chemical or enzymatic modifications. After leaving the stomach, ingested compounds reach the small intestine, in which a major part of the host metabolic enzymes are secreted, possibly leading to further enzymatic modifications. The compound, in its original or modified form, may subsequently be absorbed and enter circulation, or may further pass through the intestine. Here, food compounds may have a local biological activity by coming into contact with the complex microbial community present in the terminal ileum (last portion of the small intestine) and the colon (Alegria et al, 2015).

[0227] Human studies are certainly one of the most representative ways to study the different intestinal processes. However, they are highly labor- and time-consuming, very costly and do not allow mechanistic studies.

[0228] In humans, the intestine can be considered a black box that allows quantifying the in- and output but investigating the underlying intestinal processes in their different compartments is difficult due to sampling issues. Moreover, ethical constraints limit the general application of human trials.

[0229] Therefore, well-designed in vitro simulation technologies offer a very useful alternative for human and animal studies. Being representative for specific processes, such models enable reproducible in-depth studies of these processes without ethical constraints. The easier setup and sampling allow medium to high throughput studies at lower costs. Nonetheless, the lack of a physiological host environment is the most important limitation of these models. However, the use of standardized in vitro cell cultures using human-derived cell lines provide a fast and reproducible way to study the ultimate effects of compounds on the intestinal mucosa. Furthermore, extensive in vitro investigation allows to carefully design subsequent animal or human studies, thereby saving time and money.

[0230] Carefully designed GIT-simulating in vitro approaches offer an excellent high-throughput screening setup to evaluate the putative metabolic fate of selected food ingredients at different concentrations. Such ingredients may be modified or modify the bacterial community in the gut, and therefore, reach the intestinal mucosa intact or in the form of a modified by-product. Oral bioavailability of dietary compounds is defined as the fraction of the administered dose able to be absorbed by the intestinal cells and that is available for use or storage.

[0231] Bioavailability of dietary compounds is dependent on many factors, namely on the nutritional and physiological status of the individual, on the conjugation of the compound with other nutrients and/or bile salts, on the enzymatic degradation of the compound by digestive enzymes, and on the capacity of the gut-associated bacteria to metabolize it. In vitro gastrointestinal model offers the possibility to screen a large set of molecules in a rapid and cost-effective way in short-term experiments.

[0232] The following approach allows to rapidly assess the intestinal fate of dietary compounds upon digestion and colon fermentation. This, associated with in vitro cell models that mimic the human intestinal epithelium, allow to investigate the bioavailability of the intact and modified compounds, thereby increasing both the scientific output and commercial relevance.

[0233] Firstly, short-term screening assays were performed as a tool to evaluate the digestive fate of curcumin-based formulations, with different solubility properties.

[0234] The results of these experiments were then applied to Caco-2 cells in vitro in order to investigate the bioavailable fraction of the different formulations, in comparison to their unmodified/undigested forms.

[0235] In addition, cellular toxicity was measured in order to compare the putative cytotoxic effects of the different digestive fractions.

[0236] The short-term screening assays consisted of the sequential incubation (stomach, small intestine, colon) of a representative dose(s) of the selected lead compounds under simulated conditions for the large intestine with a representative bacterial inoculum.

[0237] An intestinal suspension collected from the ascending colon compartment of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) was used (Van de Wiele et al, 2015).

[0238] This inoculum consists of a stable microbial community which is adapted, both in structure and activity, to the environmental conditions which are present in the proximal colon.

[0239] The following curcumin-based formulations/composition were evaluated:

[0240] 1. A standard curcumin extract (Curcuma longa)—containing a mix of 3 curcuminoids (curcumin—75%, demethoxicurcumin (DMC)—15 to 20% and bidemethoxycurcumin (BDMC)—5 to 10%).

[0241] 2. A control formulation (Turmeric phytosome Thorne product with Meriva® which formulation comprises 18-22% curcuminoids, where the curcumin and soy lecithin present are formulated in a 1:2 weight ratio (Phytosome), and two parts of microcrystalline cellulose are then added to improve flowability, with an overall content of curcumin in the final product of around 20%.)

[0242] 3. A composition as used in the methods/uses of the invention comprising 8.6% turmeric extract (with more than 6% curcuminoids), 15.9% sunflower oil, 2% quillaja extract, and 73.5% modified starch) (also referred to herein as Form I).

[0243] The short-term screening assay consisted on the sequential incubation of the three formulations under stomach, small intestine and colon conditions.

[0244] The formulations/compositions were tested to achieve a curcuminoids concentration of 0.5 g/L in the stomach compartment (the actual amount in mg was calculated based on the % of curcuminoids within each product—as shown in Table 2).

[0245] The different formulations/compositions were then incubated for 1 hour (h) at 37° C., pH 2.0, in the presence of pepsin.

[0246] The small intestine was then simulated by adding pancreatic enzymes and bile salts and samples were incubated at 37° C. for a total duration of 3 h.

[0247] Finally, in the third incubation stage, the colon was simulated by adding a representative fecal inoculum collected from the SHIME and a rich nutritional media. Colon incubations were carried out at 37° C., with shaking and under anaerobiosis, over a total duration of 48 h.

[0248] Each formulation/composition was tested in triplicate to control for biological variability.

[0249] Note that these experiments were designed in order to respect the specific residence times of food ingredients in the gastrointestinal tract. Considering the volumes within each compartment, the concentration of curcuminoids tested was: 0.5 g/L in the stomach, 0.35 g/L in the small intestine and 0.1 g/L in the colon. For the cell transport experiments these samples were diluted 10× more.

TABLE-US-00002 TABLE 2 Curcumin-based formulations/compositions tested and the respective curcuminoids percentage in the formulation/composition. The curcuminoids percentage was taken into account in order to calculate the amount in mg to be added to the stomach compartment. In bold is depicted the native form. Curcuminoids content % Description Curcumin DMC BDMC Total Standard Turmeric powder 79.42 14.72 2.03 96.18 extract (95% curcuminoids) Turmeric phytosome (Thorne 13.43 2.70 0.28 16.40 product with Meriva ®) A composition as used in the 5.50 0.77 0.08 6.35 methods/uses of the invention comprising turmeric extract, sunflower oil, quillaja extract and modified starch

[0250] Samples for each formulation/composition were collected at the following time points: [0251] Stomach: 30 min and 60 min [0252] Small intestine: 60, 120 and 180 min [0253] Colon: 2, 4, 6, 24 and 48 hours

[0254] The samples were then analyzed for their curcuminoids content (curcumin, DMC and BDMC) using high pressure liquid chromatography (HPLC) coupled with mass spectrometry.

[0255] A calibration curve was prepared in the range 2-1000 ng/mL for each 3 curcuminoids (Phytolab, Vestenbergsgreuth, Germany) adding 54 ppb of curcumin-d6 (TLC pharmachem, Ontario, Canada) as an internal standard to ensure retention time stability and instrument correction variation. Acetonitrile was used as the diluent for each solution. For free curcuminoid determination, exactly 450 μL of internal standard solution (60 ng/mL) was loaded over 50 μL of plasma sample into Captiva 96 wells plate (ND lipids from Agilent). After mixing and filtration the eluate is ready to be injected into LC/MS system. Captiva ND Lipid plates are designed to effectively remove phospholipids from plasma. For the determination of total conjugated curcuminoid metabolites (glucuronide and sulfate metabolites), 100 μL of plasma sample was mixed with 100 μL of enzyme solution (either glucuronidase 1000 units/mL, Sigma #G7017; or sulfatase, Sigma #S9626, 100 units/mL) for 2 hours at 37° C. After this hydrolysis step, 50 μL of the solution is mixed with 450 μL of acetonitrile onto Captiva 96 wells plate as well. The sample procedure is the same than for free curcuminoids, mixing and filtering before injection.

[0256] LC/MS conditions were then as follows. The autosampler (5° C.) and LC system used was an Agilent Infinity 1290 integrated system. Agilent 6420 Triple quadrupole mass spectrometer was used during the study, with electrospray ionization. The metabolites were eluted from the BEH Shield RP 18 column (100×2.1, 1.7 μm; Waters) with a mobile phase consisting of 0.1% formic acid in water in HPLC grade (solvent A) and 0.1% formic acid in acetonitrile (solvent B), at a flow rate of 0.5 mL/min. The elution was in gradient from 40-80% B at 0-6 min. The injection volume was 2 μL for standard and samples. For each reference compound, a relevant transition of the precursor-to-product ions were detected with the utilization of the multiple reaction monitoring (MRM) mode. For each of the 3 analytes was determined in MS1 full scan tests and the product ions in MS/MS experiments. MRM transitions of each analyte were optimized using direct infusion and Optimizer B.08.00 workstation software solution (Agilent technologies, Santa Clara, Calif., USA). See Table 2a for the optimal selected conditions. The mass spectrometer parameters were set as follows: ESI source both in negative and positive mode; drying gas (N2) flow rate, 10 L/min; gas temperature, 350° C.; nebulizer, 40 psi; and capillary, 4.0 kV. The MS system fully calibrated prior to running according to manufacturer's guidelines. Data analysis were carried out on Agilent MassHunter Quantitative/Qualitative analysis B.07.00 (Agilent technologies, Santa Clara, Calif., USA).

TABLE-US-00003 TABLE 2a Retention times (Tr), multiple reaction monitoring (MRM) transitions, and optimized tandem mass spectrometry (MS/MS) detection parameters of 3 curcuminoids and internal standard. Prec. Prod. Ion Q1 ions Q3 Dwell Rt Mass Mass Time Frag CE Compounds (min) ISTD? (Da) (Da) (ms) (V) (V) Polarity Curcumine 3.25 yes 375.2 291.1 50 110 12 positive d6 180.1 110 18 Curcumine 3.29 No 369.1 285.1 98 13 177.1 98 25 DMC 3.49 No 339.1 255.1 110 12 177.1 110 16 BDMC 3.68 No 309.1 225.1 110 12 119.1 110 36

[0257] Table 3 depicts the concentration in curcuminoids obtained among the intestinal tract compartments during digestion.

[0258] Surprisingly the concentrations after digestion in the stomach after 60 min and the small intestine after 120 min and 180 min for the composition as used in the methods/uses of the invention were superior than the ones obtained for the standard turmeric powder extract, demonstrating a better resistance to digestion for the composition as used in the methods/uses of the invention in comparison to the standard extract. Also, the concentrations after digestion in the stomach after 60 min and the small intestine after 120 min and 180 min for the composition as used in the methods/uses of the invention were higher than the ones of the comparator Turmeric phytosome formulation, demonstrating a better resistance to digestion for the composition as used in the methods/uses of the invention in comparison to the Turmeric phytosome comparator.

[0259] Table 4 shows the percentage of curcuminoids that remained in the intestinal compartments after digestion according to the initial concentrations (0.5 g/l or 500 mg/l of curcuminoids in the stomach compartment at the beginning of the experiments). The results clearly show that surprisingly, curcuminoids coming from the composition as used in the methods/uses of the invention had a far better resistance to digestion in the stomach and small intestine in comparison to curcuminoids coming from the standard turmeric extract and composition as used in the methods/uses of the invention are more protected from degradation during digestion after oral consumption then curcuminoids from the standard extract or the Turmeric phytosome formulation and that there are much more curcuminoids accessible for absorption in the small intestine compartment after oral consumption of the composition as used in the methods/uses of the invention than oral consumption of the standard extract or the turmeric phytosome formulation.

TABLE-US-00004 TABLE 3 Concentration of curcuminoids in the different compartments of the gastro-intestinal tract during the digestion process for the three formulations Concentration Turmeric phytosome (Thorne The composition as used in the after digestion Standard Turmeric powder extract product with Meriva ®) methods/uses of the invention (ppm or mg/l) Curcumin DMC BDMC Total Curcumin DMC BDMC Total Curcumin DMC BDMC Total Initial 412.915 76.511 10.573 500.000 409.451 82.165 8.384 500.000 433.018 60.678 6.304 500.000 412.915 76.511 10.573 500.000 409.451 82.165 8.384 500.000 433.018 60.678 6.304 500.000 412.915 76.511 10.573 500.000 409.451 82.165 8.384 500.000 433.018 60.678 6.304 500.000 ST60 min 239.880 45.610 6.523 292.013 ND 398.600 81.901 11.880 492.381 162.430 31.080 5.108 198.618 345.510 71.428 10.983 427.921 125.390 27.570 4.564 157.524 426.930 88.494 12.595 528.019 Mean 175.900 34.753 5.399 216.052 390.347 80.608 11.819 482.774 SD 58.421 9.565 1.011 68.919 41.333 8.606 0.808 50.736 SI 120 min 36.417 8.723 3.772 48.912 107.857 23.464 5.250 136.571 325.371 58.626 11.509 395.506 53.372 11.500 4.407 69.279 108.571 23.957 5.200 137.729 342.014 66.726 10.613 419.353 39.145 9.041 3.940 52.126 109.286 31.286 6.957 147.529 312.314 61.350 9.924 383.588 Mean 42.978 9.755 4.040 56.772 108.571 26.236 5.802 140.610 326.567 62.234 10.682 399.482 SD 9.104 1.520 0.329 10.950 0.714 4.380 1.000 6.020 14.886 4.122 0.795 18.211 SI 180 min 36.188 8.150 3.699 48.037 110.000 24.564 5.057 139.621 276.500 54.060 9.117 339.677 34.933 8.500 3.861 47.294 110.714 28.243 5.793 144.750 236.500 46.564 8.105 291.169 40.275 9.211 4.433 53.919 61.329 29.569 6.604 97.501 301.729 58.411 10.343 370.483 Mean 37.132 8.620 3.998 49.750 94.014 27.459 5.818 127.291 271.576 53.012 9.188 333.776 SD 2.793 0.541 0.386 3.629 28.309 2.593 0.774 25.926 32.892 5.993 1.121 39.985 Colon 2 h 15.623 11.508 9.989 37.119 7.521 6.810 2.280 16.611 5.248 3.554 1.647 10.449 12.307 9.337 7.965 29.609 8.137 9.334 4.403 21.873 5.783 3.660 1.497 10.939 11.896 11.535 11.990 35.420 6.263 6.628 2.742 15.632 4.820 3.443 1.731 9.994 Mean 13.275 10.793 9.981 34.049 7.307 7.590 3.141 18.039 5.284 3.552 1.625 10.461 SD 2.043 1.261 2.013 3.938 0.955 1.512 1.116 3.357 0.482 0.108 0.119 0.473 Colon 24 h 11.364 5.136 1.785 18.284 7.815 4.405 1.171 13.391 4.408 1.744 0.551 6.703 10.001 4.828 1.820 16.649 7.187 4.392 1.154 12.732 4.427 1.788 0.569 6.783 9.297 4.358 1.249 14.903 0.896 0.734 0.218 1.847 4.073 1.460 0.292 5.824 Mean 10.220 4.774 1.618 16.612 5.299 3.177 0.847 9.323 4.303 1.664 0.470 6.437 SD 1.051 0.392 0.320 1.691 3.826 2.116 0.545 6.483 0.199 0.178 0.155 0.532 Colon 48 h 6.469 2.303 0.549 9.320 5.158 2.926 0.727 8.811 2.453 0.870 0.199 3.521 4.992 2.403 0.697 8.091 3.750 2.423 0.614 6.786 2.976 1.197 0.303 4.476 6.383 2.858 0.837 10.077 4.438 2.488 0.609 7.535 3.285 1.192 0.315 4.792 Mean 5.948 2.521 0.694 9.163 4.448 2.612 0.650 7.711 2.905 1.086 0.272 4.263 SD 0.829 0.296 0.144 1.002 0.704 0.274 0.067 1.024 0.420 0.188 0.064 0.661 Each formulation was tested in triplicate; the results present the mean and SD of the 3 experiments. DMC: Demethoxycurcumin; BDMC: Bisdemethoxycurcumin; ND: Not determined; SD: Standard deviation; ST: Stomach; SI: Small Intestine

TABLE-US-00005 TABLE 4 Resistance to Gastro-intestinal (GIT) digestion: percentage of curcuminoids that remained in the intestinal compartments after digestion according to the initial concentration Turmeric The composition as Resistance to GIT phytosome (Thorne used in the digestion (% of initial Standard Turmeric product with methods/uses of concentration) powder extract Meriva ®) the invention Stomach 60 min 43.21 ± 13.78% ND 96.55 ± 10.15% SI 120 min 11.35 ± 2.19%  28.12 ± 1.20%  79.90 ± 3.64%  SI 180 min 9.9 5 ± 0.73%  25.46 ± 5.19%  66.76 ± 8.00%  Colon 2 h 6.81 ± 0.79% 3.61 ± 0.67% 2.09 ± 0.09% Colon 24 h 3.32 ± 0.34% 1.86 ± 1.30% 1.29 ± 0.11% Colon 48 h 1.83 ± 0.20% 1.54 ± 0.20% 0.85 ± 0.13% The data shows mean ± SD of the percentage of curcuminoids concentration in the different compartments and the different times after digestion in comparison to the initial concentration.

[0260] The following samples were also collected for the transport experiments on Caco-2 cells: [0261] Small intestine: 120 and 180 min

[0262] The pH of the samples to be applied to the cells was adjusted to 6.5 prior to use.

[0263] Caco-2 cells are widely used as a cellular model for intestinal function, as they are able to spontaneously differentiate into enterocyte-like cells in culture. When cultured in semi-permeable supports, these cells develop into a functional polarized monolayer that resembles the intestinal epithelia, with the presence of apical brush-border enzymes and microvilli. Therefore, because they acquire in culture morphological and functional characteristics of mature enterocytes, they are considered as the “gold standard” model for transport experiments (Sambuy et al, 2002).

[0264] Caco-2 cells (ATCC) were seeded in 12-transwell inserts (0.4 μm) at a density of 0.9×105 cells/cm.sup.2, corresponding to 1×105 cells/insert. Cells were let differentiate until a functional monolayer was reached (21 days); the apical (600 μL) and the basolateral (1500 μL) media were replenished three times a week. On the day of the experiment, the barrier function was assessed by measuring the transepithelial electrical resistance (TEER) of the monolayer. Cells were washed with HBSS to remove traces of media, and 2 mL of transport buffer (TB) was added to the basolateral side. The samples collected from the short-term experiments were diluted in transport buffer at 1:10 (v/v) ratio and given apically to the cells (600 μL). All products were also tested unprocessed and the powders diluted in TB at a concentration of 0.025 mg/mL (the final theoretical concentrations of curcuminoids tested for all formulations can be seen in Table 2a). These dilutions were prepared from stock solutions (250 mg/mL) prepared in HBSS, except for the standard turmeric extract, which was dissolved in DMSO, due to its poor solubility. As control wells, we have used colon samples (48 h incubation) diluted 1:10 (v/v) in TB obtained by running during the short-term experiments a blank (no curcumin). The transport buffer (TB) consisted of HBSS (pH 7.4) supplemented with 10 mM HEPES, 25 mM D-Glucose and 1× antibiotic-antimycotic. Cells were incubated for a total duration of 4 h at 37° C.

[0265] The following samples were collected:

[0266] 1. Diluted samples that were used to stimulate the cells (500 μL). These correspond to a 0 h time point, as it contains the diluted samples before being given to the cells unprocessed formulations diluted in TB at a concentration of 0.025 mg/mL were also shipped.

[0267] 2. Samples from the apical side collected after 2 h and 4 h of incubation (250 μL each).

[0268] 3. Samples from the basolateral side collected after 2 h (800 μL) and 4 h (1000 μL) of incubation.

[0269] 4. Samples from the cells after 4 h of incubation. These correspond to the fraction which has been uptaken inside the cells. Briefly, ice-cold PBS 1× was added to the cells to terminate the transport. Then, the cells were washed once more with PBS 1× to remove traces of product which has not been internalized, and cells were permeabilized with a solution of PBS 1× containing 20% of ethanol and 0.1% Tween-20 (600 μL); after 20 min in this solution, the cells were collected into a 1.5 mL tube and disrupted and homogenized with the help of a syringe and a 21 G needle. The tubes were centrifuged and the supernatant transferred into a new tube (450 μL).

[0270] All samples were stored at −20° C. until HPLC analysis.

[0271] In order to evaluate the cytotoxicity of the different samples applied to Caco-2 cells, lactate dehydrogenase (LDH) released by Caco-2 cells on the apical side (after 4 h of incubation) was evaluated by using a LDH-Activity Kit. LDH is released into the supernatant by cells upon membrane injury, and is therefore a marker for cell death.

[0272] Statistical analysis was done using a one-way ANOVA followed by a Dunnett's post-hoc multiple comparison test. (*), (**) and (***) correspond to significances at p<0.05, p<0.01 and p<0.001, respectively.

[0273] As shown in FIG. 10 to 15, the control wells (TB+colon 48 h) show a LDH activity around 1.0. As it is possible to see, for all products, both the undigested form as well as the stomach samples collected after 60 min shown the highest LDH activity when compared to the control.

[0274] In contrast, both small intestine and colon samples show levels comparable to the control or lower. These results demonstrated that except from samples coming from the stomach, all samples exhibited no toxicity on Caco-2 cells and results of the assays testing transport and bioavailability of curcuminoids from the samples in Caco-2 cells could be exploited and judged as valid because obtained on viable cells.

[0275] Samples from the apical, basolateral and intracellular compartments of Caco-2 cells incubated with either undigested products or with samples coming from the small intestine (120 min or 180 min) were further analyzed for their curcuminoids content (curcumin, DMC and BDMC) and their relative metabolites content (Curcumin sulfate, curcumine glucuronide, DMC sulfate and DMC glucuronide, BDMC sulfate and BDMC glucuronide) as Caco-2 cells are known to express UDP-Glucuronosyltransferases and sulfotransferases (Siissalo S, Zhang H, Stilgenbauer E, Kaukonen A M, Hirvonen J, Finel M. The expression of most UDP-glucuronosyltransferases (UGTs) is increased significantly during Caco-2 cell differentiation, whereas UGT1A6 is highly expressed also in undifferentiated cells. Drug Metab Dispos. 2008 November; 36(11):2331-6) therefore being able to metabolize curcumin, DMC and BDMC in their glucuronide or sulfate metabolites (Dempe J S, Scheerle R K, Pfeiffer E, Metzler M. Metabolism and permeability of curcumin in cultured Caco-2 cells. Mol Nutr Food Res. 2013 September; 57(9):1543-9).

[0276] The apparent permeability coefficient (P.sub.app) values for the apical to basolateral transition were calculated according to Artursson and Karlsson using the formula:

P.sub.app [cm/s]=(Vapi/(A*t))*(Cbaso/Capi)

where Vapi is the volume of the apical compartment (0.6 mL), A is the surface area of the monolayer (1.131 cm.sup.2), t is the time (s), Cbaso is the concentration (ppm) of the total curcuminoids and their metabolites in the basolateral compartment (sum of parent compound and metabolites), and Capi is the initial concentration (ppm) of total curcuminoids in the apical compartment.

[0277] The table 5 depicts the P.sub.app values for different time intervals after apical exposure of Caco-2 cells to the standard extract or the 2 different formulations (Turmeric phytosome or the composition as used in the methods/uses of the invention).

TABLE-US-00006 TABLE 5 P.sub.app values of total curcuminoids and their metabolites (expressed as 10.sup.−7 cm/s) calculated for different time intervals after apical exposure of Caco-2 cells to the standard extract or the 2 different formulations After 2 h After 4 h P.sub.app (×10.sup.−7 cm/s) of incubation of incubation Standard Turmeric powder 2.13 ± 0.23 1.10 ± 0.07 extract Turmeric phytosome 9.94 ± 0.20 5.33 ± 0.25 (Thorne product with (4.7) (4.8) Meriva ®) The composition as used 41.01 ± 1.37  22.33 ± 0.43  in the methods/uses of the (19.3) (20.3) invention with turmeric [4.1] [4.2] extract, sunflower oil, quillaja extract and modified starch The data shows mean ± SD. The fold increase in P.sub.app relative to standard extract is shown into brackets ( ). The fold increase in P.sub.app relative to turmeric phytosome formulation is shown in square brackets [ ].

[0278] As shown in table 5, the apparent permeability coefficient (P.sub.app) values for the apical to basolateral transition was surprisingly higher for the composition as used in the methods/uses of the invention then for the standard turmeric extract, with a fold increase of 19.3 and 20.3 in P.sub.app value relative to standard turmeric extract respectively at 2 h and 4 h.

[0279] The apparent permeability coefficient (P.sub.app) value for the apical to basolateral transition was also higher for the Turmeric phytosome formulation, which was used as a positive control for an enhanced bioavailability formulation, then for the standard turmeric extract, with a fold increase of 4.7 and 4.8 in P.sub.app relative to standard turmeric extract respectively at 2 h and 4 h. But the results show that absorption of curcuminoids by Caco-2 absorptive cells was greater for the composition as used in the methods/uses of the invention (P.sub.app value 4.1-fold and 4.2-fold superior at 2 h and 4 h respectively) in comparison to the comparator turmeric phytosome formulation.

[0280] Table 6 depicts the P.sub.app values after apical exposure of Caco-2 cells to the small intestine digestion samples (120 or 180 min) of the standard extract or the 2 different formulations (Turmeric phytosome or the composition as used in the methods/uses of the invention) using the quantified concentrations of curcuminoids in the small intestine compartments at 120 min or 180 min as Capi. It reflects the absorption capacity of curcuminoids from the digested formulation by the cells. When standard extract or formulations have been digested in the stomach and small intestine compartments and therefore samples coming from the small intestine (120 min or 180 min) are used to measure the absorption of curcuminoids by Caco-2 cells, we demonstrated that absorption of curcuminoids by Caco-2 absorptive cells was greater for the composition as used in the methods/uses of the invention (P.sub.app value 1.8-fold and 16.4-fold higher for the small intestine sample at 120 min or 180 min respectively) in comparison to the turmeric phytosome formulation.

[0281] Table 7 depicts the P.sub.app values after apical exposure of Caco-2 cells to the small intestine digestion samples (120 or 180 min) of the standard extract or the 2 different formulations (Turmeric phytosome or the composition as used in the methods/uses of the invention) using the theoretical concentrations of curcuminoids in the small intestine compartments at 120 min or 180 min as Capi in order to take into account not only the absorptive capacity of the cells but also resistance to the digestive process. We demonstrated from those data that the level of curcuminoids that can reach the basolateral compartment (that mimics blood circulation) after gastrointestinal digestion and absorption by Caco-2 cells during 120 min and 180 min are much higher for the composition as used in the methods/uses of the invention in comparison to the standard turmeric extract turmeric (P.sub.app value 2.0-fold and 1.8-fold higher for the small intestine sample at 120 min or 180 min respectively) and in comparison to the phytosome formulation (P.sub.app value 3.4-fold and 7.1-fold higher for the small intestine sample at 120 min or 180 min respectively).

TABLE-US-00007 TABLE 6 P.sub.app values of total curcuminoids and their metabolites after apical exposure of Caco-2 cells to the small intestine digestion samples (120 or 180 min) of the standard extract or the 2 different formulations After 2 h of incubation of SI digestion SI digestion P.sub.app (×10.sup.−8 cm/s) samples 120 min samples 180 min Standard Turmeric powder extract 2.42 ± 1.09 7.19 ± 7.70 Turmeric phytosome (Thorne 0.59 ± 0.08 0.19 ± 0.06 product with Meriva ®) The composition as used in the 1.08 ± 0.54 3.08 ± 1.03 methods/uses of the invention with (1.8) (16.4) turmeric extract, sunflower oil, quillaja extract and modified starch The data shows mean ± SD. The fold increase in P.sub.app relative to turmeric phytosome formulation is shown into brackets ( ). The P.sub.app values are calculated using the quantified concentrations of curcuminoids in the small intestine compartments at 120 min or 180 min as Capi.

TABLE-US-00008 TABLE 7 P.sub.app values of total curcuminoids and their metabolites after apical exposure of Caco-2 cells to the small intestine digestion samples (120 or 180 min) of the standard extract or the 2 different formulations After 2 h of incubation of SI digestion SI digestion P.sub.app (×10.sup.−9cm/s) samples 120 min samples 180 min Standard Turmeric powder extract 2.59 ± 0.80 6.90 ± 0.74 Turmeric phytosome (Thorne 1.54 ± 0.04 1.79 ± 0.02 product with Meriva ®) the composition as used in the 5.30 ± 2.44 12.65 ± 0.36 methods/uses of the invention with (2.0) (1.8) turmeric extract, sunflower oil, [3.4] [7.1] quillaja extract and modified starch The data shows mean ± SD. The fold increase in P.sub.app relative to standard extract is shown into brackets ( ). The fold increase in P.sub.app relative to turmeric phytosome formulation is shown in square brackets [ ]. The P.sub.app values are calculated using the theoretical concentrations of curcuminoids in the small intestine compartments at 120 min or 180 min as Capi.

Example 6—Test the Effect of a Composition of the Invention to Enhance the Bioavailability of Curcuminoids Through Comparative Pharmacokinetic Studies In-Vivo in Mice

[0282] In view of the results obtained in the in vitro model which showed that the composition as used in the methods/uses of the invention showed better resistance to gastrointestinal digestion and better absorption through intestinal cells, it is postulated that the composition as used in the methods/uses of the invention (i.e. a composition comprising 8.6% turmeric extract (comprising more than 6% curcuminoids), 15.9% sunflower oil, 2% quillaja extract, and 73.5% modified starch), will improve the bioavailability of curcuminoids in comparison to a standard turmeric extract in mice.

[0283] Therefore, a comparative pharmacokinetic study was conducted in mice.

[0284] Adult male C57Bl/6J Rj mice from Janvier Labs (St-Berthevin, France), at 5 weeks of age at receipt, were housed collectively in standard plastic cages (n=4/cage). All animals had ad libitum access to water and standard pellet food (pellet A04; SAFE, Villemoisson-sur-Orge, France), and were maintained in a temperature—(24.0 to 26.0° C.) and humidity-(40.0 to 50.0%) controlled room on a 12-h light (07:00 AM-07:00 PM)/12-h dark cycle.

[0285] All animals were acclimatized to their new environment for one week following receipt. In order to ensure both a correct acclimatization and a standard growth curve, the global food intake and the body weight of the animals were evaluated twice per week. Following this acclimatization period, mice were habituated to receive a daily per os administration of vehicle (Carboxymethylcellulose sodium salt 1% (w/v) dissolved in distillated water at room temperature, CMC; Ref #C4888, Batch number: SLBB5612V, SIGMA ALDRICH, St Quentin Fallavier, FRANCE) for six days before the treatment. During this habituation period, the global food intake and the body weight of the animals were measured daily. These measures allowed to be sure of an optimal habituation of the animals to both the injection procedures and the manipulation by the experimenters.

[0286] During the acclimatization and habituation periods, mice were allowed ad libitum access to a preweighed quantity of fresh food pellets (pellet A04; SAFE, Villemoisson-sur-Orge, France). The remaining food was weighed the next day of measure. Using a precision scale (THB-600G, PMC Millot; precision±0.01 g), the global food intake per cage (08:40-09:20 AM) was determined by subtracting the remaining food to the preweighed quantity of food. The mean daily food intake was obtained by dividing this value by the number of days separating two measures and by the number of animals of each cage. At each body weight measurement, mice were weighed in the morning (08:40-09:20 AM).

[0287] The day prior the treatment (last day of the habituation period), 120 mice were fasted in the evening (17:40-18:20 PM). The day after, mice (n=40 per formulation) received an acute treatment by oral gavage (30 ml/kg) in the morning (08:00-09:50 AM) with a standard turmeric powder extract, the Tumeric phytosome formulation as a comparator or the composition as used in the methods/uses of the invention (i.e. a composition comprising 8.6% turmeric extract (with more than 6% curcuminoids), 15.9% sunflower oil, 2% quillaja extract, and 73.5% modified starch).

[0288] An appropriate volume of Vehicle (CMC 1% dissolved in distilled water) was added in a suitable recipient under agitation and the pH was adjusted to 5.5. An appropriate amount of pre-weighted formulation was gradually added to the vehicle under constant agitation. Once homogeneous, the pH of the obtained suspension was measured and adjusted 5.5 if needed. In order to avoid any degradation of the curcuminoids, the final suspension was systematically administered in 1 h following preparation. Doses were calculated to fed animals with 300 mg/kg of total curcuminoids (see table 8). This dosage in mice is equivalent to 21.37 mg/kg in humans and 1282 mg assuming a 60 kg human—formula from (U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER). Guidance for Industry Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers. 2005); Human Equivalent Dose (mg/kg)=animal dose in mg/kg×(animal weight in kg/human weight in kg).sup.0.33. Doses were adjusted to the body weight measured just before fasting. In order to avoid any impact of feeding and drinking behavior on intestinal absorption after oral gavage with the formulation, animals were water- and food-deprived during the first 12 h following the treatment.

TABLE-US-00009 TABLE 8 Curcuminoids content of the standard turmeric extract and the 2 formulations and the respective concentration of the suspension used for product administration to mice at 300 mg/kg body weight of curcuminoids in the first in vivo study Curcuminoids C. ° of dosing content solution (mg of Formulation (g/100 g) formulation/ml) Standard Turmeric powder 96.18 10.40 extract (95% curcuminoids) Turmeric phytosome 16.40 60.98 (Thorne product with Meriva ®) The composition as used in 6.345 157.60 the methods/uses of the invention with turmeric extract, sunflower oil, quillaja extract and modified starch

[0289] Blood was sampled at 0.5-, 1 h-, 2 h-, 4 h-, 6 h-, 8 h-, 12 h- or 24 h-post-dosing by cardiac puncture on anaesthetized mice (n=5/time point/formulation). Anesthesia was performed by intraperitoneal injection of a mixture of Ketamine/Xylazine (100 mg/kg and 15 mg/kg, respectively). For cardiac puncture, a 26 G syringe was inserted between the eighth and the tenth sternal rib, with an angle of 450 with the longitudinal axis formed by the body of the animal, in order to directly penetrate the left heart ventricle. Blood was then gently drawn to obtain a final volume of 0.6-1 ml. For the interests of the bioanalytical method, blood was thereafter transferred into an Eppendorf tube, mixed with heparin sulfate (200 U.Math.l/ml of blood) and gently agitated. All samples were centrifuged during 15 minutes at 3000 g and 4° C. within 30 minutes after blood collection to isolate plasma. Plasma (supernatant) was aliquoted in a new 0.5 ml Eppendorf. The aliquots of plasma were frozen at −80° C. within 1 hour after centrifugation.

[0290] Plasmatic dosages of parent curcuminoids (curcumin, DMC or BDMC) and their relative metabolites (Curcumin glucuronide and sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) were performed by a LC-MS-MS method. A calibration curve was prepared in the range 2-1000 ng/mL for each 5 curcuminoids (Phytolab, Vestenbergsgreuth, Germany) adding 54 ppb of curcumin-d6 (TLC pharmachem, Ontario, Canada) as an internal standard to ensure retention time stability and instrument correction variation. Acetonitrile was used as the diluent for each solution. For free curcuminoid determination, exactly 450 μL of internal standard solution (60 ng/mL) was loaded over 50 μL of plasma sample into Captiva 96 wells plate (ND lipids from Agilent). After mixing and filtration the eluate is ready to be injected into LC/MS system. Captiva ND Lipid plates are designed to effectively remove phospholipids from plasma. For the determination of total conjugated curcuminoid metabolites (glucuronide and sulfate metabolites), 100 μL of plasma sample was mixed with 100 μL of enzyme solution (either glucuronidase 1000 units/mL, Sigma #G7017; or sulfatase, Sigma #S9626, 100 units/ml) for 2 hours at 37° C. After this hydrolysis step, 50 μL of the solution is mixed with 450 μL of acetonitrile onto Captiva 96 wells plate as well. The sample procedure is the same than for free curcuminoids, mixing and filtering before injection.

[0291] LC/MS conditions were then as follows. The autosampler (5° C.) and LC system used was an Agilent Infinity 1290 integrated system. Agilent 6420 Triple quadrupole mass spectrometer was used during the study, with electrospray ionization. The metabolites were eluted from the BEH Shield RP 18 column (100×2.1, 1.7 μm; Waters) with a mobile phase consisting of 0.1% formic acid in water in HPLC grade (solvent A) and 0.1% formic acid in acetonitrile (solvent B), at a flow rate of 0.5 mL/min. The elution was in gradient from 40-80% B at 0-6 min. The injection volume was 2 μL for standard and samples. For each reference compound, a relevant transition of the precursor-to-product ions were detected with the utilization of the multiple reaction monitoring (MRM) mode. For each of the 5 analytes was determined in MS1 full scan tests and the product ions in MS/MS experiments. MRM transitions of each analyte were optimized using direct infusion and Optimizer B.08.00 workstation software solution (Agilent technologies, Santa Clara, Calif., USA). See Table 9 for the optimal selected conditions. The mass spectrometer parameters were set as follows: ESI source both in negative and positive mode; drying gas (N2) flow rate, 10 L/min; gas temperature, 350° C.; nebulizer, 40 psi; and capillary, 4.0 kV. The MS system fully calibrated prior to running according to manufacturer's guidelines. Data analysis were carried out on Agilent MassHunter Quantitative/Qualitative analysis B.07.00 (Agilent technologies, Santa Clara, Calif., USA).

[0292] For the three formulations/compositions tested, the kinetic of plasma concentration of each curcuminoid compound was determined between 0.5 and 12 h post-treatment by calculating the mean±SEM plasmatic concentration at each time point of blood collection. The pharmacokinetic parameters T1/2 (half-life), Cmax, Tmax, AUC(0-12 h) and AUC(0-∞) were determined from the 0-12 h kinetic by a non-compartmental analysis using PKSolver. PKSolver is a menu-driven add-in program for Microsoft Excel written in Visual Basic for Applications (VBA), for solving problems in pharmacokinetic (Zhang et al., 2010). The whole data are represented as mean±SEM. Statistical analyses were performed with the Statview 5.0.1 (Statview software, Cary, N.C., USA) and the Excel 2013 programs. Data were analyzed by a Student's t-test at each time point. The risk a was fixed at 0.05.

TABLE-US-00010 TABLE 9 Retention times (Tr), multiple reaction monitoring (MRM) transitions, and optimized tandem mass spectrometry (MS/MS) detection parameters of curcuminoids, tetrahydrocurcumin and hexahydrocurcumin and internal standard. Prec. Ion Prod ions. Dwell Rt Q1 Mass Q3 Mass Time Frag CE Compounds (min) ISTD? (Da) (Da) (ms) (V) (V) Polarity HHC 1.07 No 373.2 179 200 118 12 negative THC 2.04 No 371.1 235.1 200 100 10 negative 193.2 100 21 Curcumine 3.25 yes 375.2 291.1 50 110 12 positive d6 180.1 110 18 Curcumine 3.29 No 369.1 285.1 98 13 177.1 98 25 DMC 3.49 No 339.1 255.1 110 12 177.1 110 16 BDMC 3.68 No 309.1 225.1 110 12 119.1 110 36

[0293] During the two successive periods of acclimatization (J1 to J7) and habituation (J8 to J14), the 24 h-food intake and the body weight of mice were regularly measured in order to ensure both a correct acclimatization and a standard growth curve before treatment. For the present study, mice were housed at four per cages, each of them being used for blood sampling at a same time point after treatment with one of the three formulations. As a consequence, the food intake and body weight data were first analyzed per group of 15 mice that have been sacrificed at a same time point (8 groups; 5 mice/time point/formulation). FIGS. 16 and 17 respectively depicts food intake and body weight of mice during the acclimatization and habituation period before treatment administration; the different groups showed classical body weight curve and food intake before treatment. It should be noted that the strong decrease in body weight observed in all groups at J15 (FIG. 17) resulted from the overnight fasting performed the night before treatment. These results confirmed that all mice used in this experiment had the same behavior and could be compared as expected.

[0294] FIG. 18 depicts the pharmacokinetic profile with concentration of the total curcuminoids (sum of curcumin, DMC, BDMC and their relative metabolites curcumin glucuronide sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) obtained at each time point in mice after oral administration (300 mg/kg bw of curcuminoids) of the Turmeric phytosome formulation and the standard extract formulation. The plasma concentration of total curcuminoids for the standard turmeric extract only reached a maximum of 12.9 ppm at 1 h and was below 10 ppm at all other time points although the turmeric phytosome formulation enabled plasma concentration of total curcuminoids to reach 41.5 ppm (μg/ml) after 30 min and was significantly superior to the one obtained for the standard at each time point except 24 h. Turmeric phystosome demonstrated a 3.2-fold increase in total curcuminoids Cmax, a 3.9-fold increase in AUC compared to standard turmeric extract. These results validated the use of the Turmeric phytosome formulation to enhance the bioavailability of curcuminoids and as a positive control and validated the relevance of our in vivo model to test the capacity of different formulations to enhance the bioavailability of curcuminoids against a standard turmeric extract.

[0295] A table containing the Mean values±SEM for each time point shown as Table 10. The numbers in brackets located next to these values indicate the number of sample which presented a positive value on the total number of samples. The result of statistical comparisons are also shown in the same table. Table 11 contains the PK parameters obtained from the non-compartmental analysis using PKSolver software. The percentage of variation between groups is also indicated (% Var°). Data are represented as mean±SEM.

TABLE-US-00011 Total curcuminoids plasmatic levels - Mean ± SEM Standard extract Turmeric phytosome P 0.5 h 9.71 ± 1.56 (5/5) 41.53 ± 7.69 (5/5) 0.0037   1 h 12.95 ± 1.56 (5/5) 34.00 ± 4.28 (5/5) 0.0022   2 h 7.10 ± 1.56 (4/4) 24.69 ± 2.41 (5/5) 0.0005   4 h 4.33 ± 1.56 (4/4) 14.31 ± 1.12 (5/5) 0.0003   6 h 3.22 ± 1.56 (5/5) 11.59 ± 1.55 (5/5) 0.0007   8 h 2.63 ± 1.56 (5/5) 15.03 ± 3.72 (5/5) 0.0110  12 h 0.55 ± 1.56 (4/4) 6.86 ± 1.80 (5/5) 0.0177  24 h 1.30 ± 1.56 (4/4) 1.44 ± 0.63 (5/5) 0.8709

TABLE-US-00012 PK parameters - Total curcuminoids Standard extract Turmeric phytosome % Var° t½ (h) 2.688208 4.926631161 183.2682 Tmax (h) 1 0.5 50 Cmax (ppm) 12.95494767 41.52686492 320.5483 AUC 0-t 49.29857342 193.9071934 393.3323 (ppm × h) AUC 0-inf 51.41308527 242.667998 471.9966 (ppm × h)

[0296] Tables 10 and 11: Showing the Mean values±SEM and the PK parameters obtained from the non-compartmental analysis using PKSolver software for each time point shown in FIG. 18.

[0297] FIG. 19 depicts the pharmacokinetic profile with concentration of the total curcuminoids (sum of curcumin, DMC, BDMC and their relative metabolites curcumin glucuronide sulfate, DMC glucuronide and sulfate, BDMC glucuronide and sulfate, THC, THC glucuronide and sulfate, HHC, HHC glucuronide and sulfate) obtained at each time pointin mice after oral administration (300 mg/kg bw of curcuminoids) of the mixture comprising curcumin, quillaja, oil and modified starch (Example 2 form 1) and the standard extract formulation. The results showed that the composition as used in the methods/uses of the invention could increase significantly total curcuminoids concentration from 0.5 h to 24 h in comparison to standard turmeric extract. The composition as used in the methods/uses of the invention demonstrated a 1.8-fold increase in total curcuminoids Cmax and a 2.2-fold increase in AUC.

[0298] A table containing the Mean values±SEM for each time point is shown as Table 12. The numbers in brackets located next to these values indicate the number of sample which presented a positive value on the total number of samples. The result of statistical comparisons are also shown in the same table. A table containing the PK parameters obtained from the non-compartmental analysis using PKSolver software is presented Table 13. The percentage of variation between groups is also indicated (% Var°). Data are represented as mean±SEM.

TABLE-US-00013 Total curcuminoids plasmatic levels - Mean ± SEM Standard extract Test formulation P 0.5 h 9.71 ± 1.56 (5/5) 18.90 ± 2.45 (5/5) 0.0133   1 h 12.95 ± 1.56 (5/5) 22.90 ± 2.45 (5/5) 0.0171   2 h 7.10 ± 1.56 (4/4) 19.90 ± 2.45 (5/5) 0.0093   4 h 4.33 ± 1.56 (4/4) 8.49 ± 2.45 (5/5) 0.0302   6 h 3.22 ± 1.56 (5/5) 8.67 ± 2.45 (5/5) 0.0012   8 h 2.63 ± 1.56 (5/5) 5.11 ± 2.45 (5/5) 0.1123  12 h 0.55 ± 1.56 (4/4) 1.86 ± 2.45 (5/5) 0.3241  24 h 1.30 ± 1.56 (4/4) 3.36 ± 2.45 (4/4) 0.1759

TABLE-US-00014 PK parameters ± Total curcuminoids Standard extract Test formulation % Var° t½ (h) 2.688208 2.70737411 100.713 Tmax (h) 1 1 100 Cmax (ppm) 12.95494767 22.89919816 176.7603 AUC 0-t 49.29857342 109.8465993 222.819 (ppm × h) AUC 0-inf 51.41308527 117.1097277 227.7819 (ppm × h)

[0299] Tables 12 and 13: Showing the Mean values±SEM and the PK parameters obtained from the non-compartmental analysis using PKSolver software for each time point shown in FIG. 19.

[0300] When looking at parent compounds specifically (curcumin, DMC and BDMC in their native form, i.e. unmetabolized), as shown in Table 14 that gives plasma concentration of parent curcuminoids for each time point after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention, the composition as used in the methods/uses of the invention was the only one for which we were able to quantify a detectable amount of parent curcuminoids during the first 4 h post-dosing and therefore to calculate the AUC(0-12 h) and AUC(0-∞) (Table 15). A 10.9-fold increase in Cmax for parent curcuminoids were obtained for the composition as used in the methods/uses of the invention in comparison to the standard turmeric extract.

[0301] When looking at parent curcumin specifically (curcumin in its native form, i.e. unmetabolized), as shown in Table 16 that gives plasma concentration of parent curcumin for each time point after consumption of 300 mg/kg from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention, the composition as used in the methods/uses of the invention was the only one for which we were able to quantify a detectable amount of parent curcumin during the first 4 h post-dosing and therefore to calculate the AUC(0-12 h) and AUC(0-∞) (Table 16). A 521.8-fold increase in Cmax for parent curcumin was obtained for the composition as used in the methods/uses of the invention in comparison to the standard turmeric extract. Moreover, the composition as used in the methods/uses of the invention induced higher plasmatic level of parent curcumin than the turmeric phytosome formulation (1.8-fold increase in Cmax).

[0302] It can be concluded from this first in vivo experiment that the composition as used in the methods/uses of the invention with more than 6% curcuminoids, obtained with 8.6% turmeric extract, 15.9% sunflower oil, 2% quillaja extract, and 73.5% modified starch and prepared according to Form 1, is able to enhance the bioavailability of total curcuminoids and their metabolites but also the bioavailability of parent compounds in comparison to a standard turmeric extract.

[0303] It can be also concluded that the composition as used in the methods/uses of the invention could better improve the bioavailability of native curcumin than the turmeric phytosome formulation.

[0304] The composition as used in the methods/uses of the invention therefore represents an attractive way to enhance parent curcumin bioavailability without using soy-derived lecithin in the formulation as opposed to the Turmeric phytosome formulation. Also, as curcumin is considered as one of the most powerful active of turmeric in comparison to DMC and BDMC and their relative reduced, glucuronide or sulfate metabolites (Ireson C, Orr S, Jones D J, Verschoyle R, Lim C K, Luo J L, Howells L, Plummer S, Jukes R, Williams M, Steward W P, Gescher A. Characterization of metabolites of the chemopreventive agent curcumin in human and rat hepatocytes and in the rat in vivo, and evaluation of their ability to inhibit phorbol ester-induced prostaglandin E2 production. Cancer Res. 2001 Feb. 1; 61(3):1058-64; Anand P, Thomas S G, Kunnumakkara A B, Sundaram C, Harikumar K B, Sung B, Tharakan S T, Misra K, Priyadarsini I K, Rajasekharan K N, Aggarwal B B. Biological activities of curcumin and its analogues (Congeners) made by man and Mother Nature. Biochem Pharmacol. 2008 Dec. 1; 76(11):1590-611; Pal A, Sung B, Bhanu Prasad B A, Schuber P T Jr, Prasad S, Aggarwal B B, Bornmann W G. Curcumin glucuronides: assessing the proliferative activity against human cell lines. Bioorg Med Chem. 2014 Jan. 1; 22(1):435-9), the composition as used in the methods/uses of the invention represents a good solution to improve the biological efficacy of curcumin for different health conditions like joint health, inflammation, arthritis, atherosclerosis, liver steatosis, liver fibrosis, diabetes, cognition, mild cognitive impairment, irritable bowel syndrome.

TABLE-US-00015 TABLE 14 concentration of parent curcuminoids (sum of curcumin, DMC and BDMC) for each time point after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the first in vivo study. Parent curcuminoids plasmatic levels - Mean ± SEM The composition as used in the methods/uses of the Standard extract Turmeric phytosome invention 0.5 h 13.91 ± 10.05 (4/5) 13.91 ± 10.05 (4/5) 151.20 ± 96.93  (5/5)   1 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 6.95 ± 6.95 (1/5)   2 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 0.30 ± 0.30 (1/5)   4 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 2.55 ± 2.55 (1/5)   6 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5)

[0305] The numbers in brackets located next to these values indicate the number of sample which presented a positive value on the total number of samples

TABLE-US-00016 TABLE 15 PK parameters obtained from the non-compartmental analysis using PKSolver software for parent curcuminoids after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the first in vivo study PK parameters-parent curcuminoids The composition as used in the Standard Turmeric methods/uses of extract phytosome the invention t½ (h) / / 0.75 Tmax (h) 0.5 0.5 0.5 Cmax (ppb) 13.9 168.3 151.2 AUC 0-t (ppb × h) / / 83.8 AUC 0-inf (ppb × h) / / 86.5

TABLE-US-00017 TABLE 16 concentration of parent curcumin for each time point after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the first in vivo study. Parent curcumin plasmatic levels-Mean ± SEM the composition as used in the methods/uses of the Standard extract Turmeric phytosome invention 0.5 h 0.20 ± 0.20 (1/5) 59.30 ± 35.26 (4/5) 104.37 ± 64.37  (5/5)   1 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 5.83 ± 5.83 (1/5)   2 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 0.30 ± 0.30 (1/5)   4 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 2.55 ± 2.55 (1/5)   6 h 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5) 0.00 ± 0.00 (0/5)

[0306] The numbers in brackets located next to these values indicate the number of sample which presented a positive value on the total number of samples

TABLE-US-00018 TABLE 17 PK parameters obtained from the non-compartmental analysis using PKSolver software for parent curcumin after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the first in vivo study PK parameters-parent curcumin The composition as used in the Standard Turmeric methods/uses of extract phytosome the invention t½ (h) / / 0.82 Tmax (h) 0.5 0.5 0.5 Cmax (ppb) 0.2 59.3 104.4 AUC 0-t (ppb × h) / / 59.6 AUC 0-inf (ppb × h) / / / 62.6

[0307] Given the results obtained in the first in vivo study in mice, showing better bioavailability of total curcuminoids and parents curcuminoids and curcumin, we decided to test an optimized formulation with an higher curcuminoids content (12% curcuminoids) prepared according Form 2 with 14.4% turmeric extract, 26.8% sunflower oil, 2% quillaja extract, and 56.8% modified starch, for its capacity to improve the bioavailability of curcuminoids in comparison to a standard turmeric extract in a second comparative pharmacokinetic study in mice.

[0308] The same methodology (mice housing, acclimatization period, habituation period, curcuminoids and their metabolites quantification using LC/MS method) was used as described earlier in this Example, but with a higher number of animals per group and time (n=12/time point/formulation) and blood was sampled at 0.25-, 0.5-, 0.75-, 1 h-, 2 h-, or 8 h-post-dosing in order to specify the kinetic profile during the earliest phase after oral consumption (300 mg/kg bw) of curcuminoids coming from a standard turmeric extract (with 79.5, 15.0 and 3.0 g/100 g of curcumin, DMC and BDMC respectively and a total of 97.5 g curcuminoids/100 g), a Turmeric phytosome formulation (with 18.6, 2.6 and 0.2 g/100 g of curcumin, DMC and BDMC respectively and a total of 21.5 g curcuminoids/100 g) or the composition as used in the methods/uses of the invention prepared according to Form 2 (with 9.8, 1.6 and 0.2 g/100 g of curcumin, DMC and BDMC respectively and a total of 11.6 g curcuminoids/100 g).

[0309] FIG. 20 depicts total curcuminoids and metabolites concentration as a function of time after consumption of the different formulations (n=72 per formulation). The results clearly showed a significant increase of curcuminoids and metabolites concentration for the Turmeric phytosome and the composition as used in the methods/uses of the invention in comparison to the standard turmeric extract. The total curcuminoids concentration was higher 0.5 h and 0.75 h after consumption of the composition as used in the methods/uses of the invention in comparison to the Turmeric phytosome formulation, showing surprisingly the better performance of the composition as used in the methods/uses of the invention in comparison to the Turmeric phytosome formulation in terms of improvement of total curcuminoids and metabolites bioavailability.

[0310] This was confirmed while calculating the corresponding area under the curve AUC(0-8 h), AUC(0-∞) and Cmax (FIGS. 21, 22 and 23 respectively) that were 280% higher, 300% higher and 337% higher for the composition as used in the methods/uses of the invention in comparison to the standard extract respectively and 6.5% higher, 30% higher and 63% higher for the composition as used in the methods/uses of the invention in comparison to the Turmeric phytosome formulation respectively (Table 18). We also showed that curcuminoids and metabolites from the composition as used in the methods/uses of the invention were more rapidly absorbed with a 1.5-fold reduction in Tmax in comparison to standard extract (0.5 h versus 0.75 h) and a 2-fold reduction in Tmax in comparison to the Turmeric Phytosome formulation (0.5 h versus 1 h) respectively (Table 18). The results also showed surprisingly that curcuminoids and metabolites from the composition as used in the methods/uses of the invention were less rapidly excreted with a longer half-life (3.8 h versus 2.8 h) in comparison to the Turmeric Phytosome formulation.

[0311] When looking at parent compounds specifically (curcumin, DMC and BDMC in their native form, i.e. unmetabolized), as shown in Table 19, that gives the PK parameters of parent curcuminoids after consumption of 300 mg/kg curcuminoids from standard turmeric extract, Turmeric Phytosome or the composition as used in the methods/uses of the invention, the composition as used in the methods/uses of the invention was surprisingly the only one for which we could calculate the AUC(0-8 h) for parent curcumin. A 3.2-fold increase in Cmax for parent curcumin was obtained for the composition as used in the methods/uses of the invention in comparison to the standard turmeric extract. No Cmax could be calculated for curcumin as no parent curcumin could be found in plasma samples after consumption of the Turmeric phytosome formulation.

[0312] It can be concluded from this second in vivo experiment that the composition as used in the methods/uses of the invention with a higher curcuminoids content (12% curcuminoids) prepared according to Form 2 with 14.4% turmeric extract, 26.8% sunflower oil, 2% quillaja extract, and 56.8% modified starch is unexpectedly able to enhance the bioavailability of total curcuminoids and their metabolites but also parent curcumin in comparison to a standard turmeric extract and to the Turmeric phytosome formulation.

[0313] The composition as used in the methods/uses of the invention therefore represents an attractive way to enhance the bioavailability of curcuminoids without using soy-derived lecithin in the formulation as opposed to the Turmeric phytosome formulation.

TABLE-US-00019 TABLE 18 PK parameters obtained from the non-compartmental analysis using PKSolver software for total curcuminoids and metabolites after consumption of 300 mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the second in vivo study PK parameters-parent curcumin The composition as used in the Standard Turmeric methods/uses of the extract phytosome invention t½ (h) 3.98 2.76 3.8 Tmax (h) 0.75 1 0.5 Cmax (ppb) 10.83 22.38 36.47 (3.4) [1.6] AUC 0-t (ppbm × h) 31.11 81.74 (2.6) 87.27 (2.8) [1.1] AUC 0-inf (ppb × h) 41.56 95.74 (2.3) 124.36 (3.0) [1.3]

[0314] The fold increase in AUC or Cmax relative to standard extract is shown into brackets ( ). The fold increase in AUC or Cmax relative to turmeric phytosome formulation is shown in square brackets [ ].

TABLE-US-00020 TABLE 19 PK parameters obtained from the non-compartmental analysis using PKSolver software for parent curcuminoids after consumption of 300mg/kg curcuminoids from standard turmeric extract, turmeric phytosome or the composition as used in the methods/uses of the invention in the second in vivo study. Standard extract Parent compounds (ppb) Total parent PK Parameter Unity curcumin DMC BDMC compounds (ppb) t½ h Missing Missing Missing Missing Tmax h 0.25 0.25 0.25 0.25 Cmax ppb 5.55 25.03 64.43 114.65 AUC 0-t ppb * h Missing Missing 16.82 29.37 AUC 0-inf_obs ppb * h Missing Missing Missing Missing Turmeric Phytosome Parent compounds (ppb) Total parent PK Parameter Unity curcumin DMC BDMC compounds (ppb) t½ h Missing Missing Missing Missing Tmax h Missing Missing Missing Missing Cmax ppb Missing Missing Missing Missing AUC 0-t ppb * h Missing Missing Missing Missing AUC 0-inf_obs ppb * h Missing Missing Missing Missing The composition as used in the methods/uses of the Total parent invention Parent compounds (ppb) compounds PK Parameter Unity curcumin DMC BDMC (ppb) t½ h Missing Missing Missing Missing Tmax h 0.25 0.25 0.25 0.25 Cmax ppb 17.94 11.13 6.34 60.45 AUC 0-t ppb * h 10.8778375 Missing Missing 32.13 AUC 0-inf_obs ppb * h Missing Missing Missing Missing

Example 7—A Comparative Pharmacokinetic Study in Healthy Volunteers to Evaluate the Ability of a Composition of the Invention to Enhance the Bioavailability of Curcuminoids

[0315] This study had two objectives:

[0316] 1. Primary Objective

[0317] To assess plasmatic concentrations profile of total curcuminoids (curcumin, demethoxycurcumin (DMC), bisdemethoxycurcumin (DBMC) and their metabolites) on a 24 hours period after consumption of a single dose of 300 mg of composition of the invention (Turmipure GOLD™ 30% curcuminoids formulation) compared to 1500 mg Standard turmeric powder extract 95% curcuminoids.

[0318] 2. Secondary Objectives

[0319] To assess, plasmatic concentrations profiles of the following parameters, after consumption of a single dose of five studied products containing either 1425 mg (standard turmeric powder extract 95% curcumindoids, Curcumin C3 complex California Gold Nutrition), 200 mg (Curcuma Platinum MannaVital), 90 mg (of a composition of the invention (Turmipure GOLD™ 30% curcuminoids)) or 60 mg (Curcumin Cell'Innov) of active substance: [0320] Total curcuminoids; [0321] Parent compounds (curcumin, DMC, BDMC) and their metabolites: curcumin, glucuronide and sulfate; DMC glucuronide and sulfate; BDMC glucuronide and sulfate; tetrahydrocurcumin (THC) native, glucuronide and sulfate; hexahydrocurcumin (HHC) native, glucuronide and sulfate.

[0322] The study was a monocentric, randomized, cross-over and open clinical trial.

[0323] The study commenced with a screening/inclusion visit (V0) followed by 5 experimental sessions (V1 to V5) during which the studied products were consumed by subjects (one different product at each session for each randomized subject). The V1 visit, took place a maximum of 3 weeks after V0, and can also constitute the randomization visit.

[0324] Each experimental session (V1 to V5) was separated by 1 week minimum and 2 weeks maximum. During each experimental session, subjects underwent kinetic blood sampling during 8 hour periods. The last kinetic blood sample was taken the day after each experimental session, 24 hours after the beginning of the kinetic. Urine collection was also performed during these visits for biobanking.

[0325] The subjects first urination was collected the morning of each experimental visit (totality of this first urination), with additional collections at 0 to 8 hours during the kinetic blood sampling on site and 8 to 24 hours when they came home. The last urine collection was brought back the day after the experimental visit (when they came back for the last blood sample, T24H, of the kinetic blood sampling).

[0326] Standard meals were provided to volunteers for diner before each experimental session and during the all duration of each kinetic (breakfast, lunch and afternoon collation).

[0327] The end of study was the day after the last experimental session V5 (V5-24H). [0328] 30 subjects were recruited for this study, according to the following main inclusion and exclusion criteria: [0329] I1: Age between 18 and 45 years (limits included); [0330] I2: BMI between 19 and 25 kg/m.sup.2 (limits included); [0331] I3: Weight stable, within ±3 kg in the last three months; [0332] I4: With routine blood chemistry values within the normal range; [0333] I5: For women: Non-menopausal with the same reliable contraception since at least 3 cycles before the beginning of the study and agreeing to keep it during the entire duration of the study (condom with spermicidal gel and estrogen/progestin combination contraception accepted) or menopausal without or with hormone replacement therapy (estrogenic replacement therapy begun from less than 3 months excluded); [0334] I6: Non-smoking or with tobacco consumption≤5 cigarettes/day and agreeing not to smoke during all experimental session (V1 to V5); [0335] I7: Agreeing not to consume food, drink and condiment containing curcumin, or other curcuminoids (DMC, BDMC) during the all duration of the study; [0336] I8: Good general and mental health with in the opinion of the investigator: no clinically significant and relevant abnormalities of medical history or physical examination; [0337] E1: Suffering from a metabolic or endocrine disorder such as diabetes, uncontrolled or controlled thyroidal trouble or other metabolic disorder; [0338] E2: Suffering from a severe chronic disease (e.g. cancer, HIV, renal failure, ongoing hepatic or biliary disorders, chronic inflammatory digestive disease, arthritis or other chronic respiratory trouble, etc.) or gastrointestinal disorders found to be inconsistent with the conduct of the study by the investigator (e.g. celiac disease); [0339] E3: Suffering from liver diseases; [0340] E4: Current disease states that are contraindicated to subjects with dietary supplementation: chronic diarrhea, constipation or abdominal pain, Inflammatory bowel diseases (Crohn's disease or ulcerative colitis), Cirrhosis, chronic laxatives use . . . ; [0341] E5: Suffering from Irritable Bowel Syndrome (IBS) diagnosed by a medical doctor and treated with chronic medication; [0342] E6: Having medical history of current pathology which could affect the study results or expose the subject to an additional risk according to the investigator; [0343] E7: Recent gastroenteritis or food borne illness such as confirmed food poisoning (less than 1 month); [0344] E8: Who made a blood donation in the 3 months before the V0 visit or intending to make it within 3 months ahead; [0345] E9: With a low venous capital not allowing to perform kinetic of blood samples according to the investigator's opinion; [0346] E10: With a known or suspected food allergy or intolerance or hypersensitivity to any of the study products' ingredient and/or of the standard meals (gluten intolerance, celiac disease, etc.); [0347] E11: Pregnant or lactating women or intending to become pregnant within 3 months ahead; [0348] E12: Exhibiting alcohol or drug dependence; [0349] E13: On any chronic drug treatment (for example anticoagulant, antihypertensive treatment, treatment thyroid, asthma treatment, anxiolytic, antidepressant, lipid-lowering treatment, corticosteroids, phlebotonic, veino-tonic, drug with impact on blood circulation . . . ) excepting oral and local contraceptives; [0350] E14: Currently taking (and during the past 3 months) any supplementation from botanical origins; [0351] E15: Having consumed curcumin-containing food supplements (curcumin, turmeric and curry) or foods (curcumin, turmeric, E100, and curry) defined as at least 3 times per week and for 2 weeks prior to testing; [0352] E27: Control record (glycaemia, GGT, ASAT, ALAT, urea, creatinine and complete blood count) with clinically significant abnormality according to the investigator.

[0353] Five products, which are dietary supplements in shape of capsules, were tested as part of this study: [0354] 1. Standard Turmeric powder extract 95% curcuminoids 1500 mg consumed as capsules (4 capsules; 375 mg powder per capsule) (STE), [0355] 2. C3 Complex® 95% curcuminoids (1500 mg)+BioPerine® 95% piperine (15 mg) consumed as commercial product Curcumin C3 complex California Gold Nutrition (3 caps; 500 mg C3 complex powder+5 mg bioperine powder per capsule) (TEP), [0356] 3. Meriva® (1000 mg) consumed as commercial product Curcuma Platinum Mannavital 20% curcuminoids (2 caps; 500 mg powder per capsule) (PHYT), [0357] 4. Novasol® (1000 mg) consumed as commercial product Curcumin Cell innov 6% curcuminoids (2 caps; 500 mg liquid per capsule) (NOV), [0358] 5. A composition as defined herein comprising turmeric extract, sunflower oil, quillaja extract, and arabic gum, consumed as capsule (1 capsule; 300 mg powder per capsule) (TURMIPURE GOLD).

[0359] In order to ensure the healthy status of subjects and to check eligibility criteria, a blood sample was taken during V0 visit for control record analysis and pregnancy test for non-menopausal women (βhCG dosage).

[0360] The sample was taken after a physical examination and verification of eligibility criteria. A maximum of 10 mL was collected.

[0361] Measurement of blood pressure were performed at each visit during the physical examination with an electronic blood pressure monitor (Carescape Dinamap®V100). Heart Rate (HR, in bpm), Systolic Blood Pressure (SBP, in mmHg) and Diastolic Blood Pressure (DBP, in mmHg) were also assessed.

[0362] All subjects attended in a 12-hour fasted state.

[0363] In preparation for the V1 to V5 visits, after the clinical examination, a venous catheter was placed on elbow crease of the subject. This catheter allowed blood sampling for the kinetic without any additional pricks.

[0364] Kinetic sample lasted approximately 8 hours, with all the subjects staying at the clinical investigational center. Ten (10) blood sampling were taken according to the following schedule: [0365] T-10 (baseline), [0366] T15>T30>T45>T60>T90>T120>T240>T360>T480,

[0367] A margin of ±30 s was authorized for T15, 1 min for T30 and T45, 2 min for T60 and T90, ±5 min for T120 to T480.

[0368] The T0 time point corresponds to study product consumption.

[0369] The volunteer was allowed to consume his/her standard lunch about 4 hours after study product consumption (just after at T240 time-point) and standard afternoon meal about 8 hours after study product consumption. Lunch was consumed in 30 minutes maximum. Water was not permitted 1 h before and 1 h after product administration. The catheter was removed after the last time point, T480.

[0370] The volunteers were then asked to come again at the clinical investigational site, in a 12-hour fasting state, the day after the visit for the last blood sampling of the kinetic, T24H. Classic venous blood sample material was used (single prick). The biological parameters were assessed with these samplings being analyzed in plasma; thus only EDTA tubes were used (5 mL per sampling).

[0371] A calibration curve was prepared in the range 10-600 ng/mL for each 5 curcuminoids (Phytolab, Vestenbergsgreuth, Germany) adding 50 ppb of labelled curcumins (TLC pharmachem, Ontario, Canada) as an internal standard to ensure retention time stability and instrument correction variation. Acetonitrile was used as the diluent for each solution. For free curcuminoid determination, exactly 500 μL of cold methanol:aetonitrile (15:85) mixture containing internal standards solution (50 ng/mL) was loaded over 100 μL of plasma sample into Captiva 96 wells plate (EMR lipids from Agilent, see FIG. 18). After mixing and filtration the curcumins was eluted with cold water:acetonitrile (1:4) mixture and the eluate was injected into LC/MS system. Captiva EMR Lipid plates were designed to effectively remove phospholipids from plasma. For the determination of total conjugated curcuminoid metabolites (glucuronide and sulfate metabolites), 60 μL of plasma sample was mixed with 60 μL of enzyme solution (either glucuronidase at 1 mg/mL, 100 mM phosphate buffer pH 6.8 or sulfatase 10 mg/mL, 100 mM acetate buffer, pH 5.0) for 1 hours at 37° C. with constant agitation (Eppendorf, Thermomixer C, 400 rpm). After this hydrolysis step, the protocol was the same as for free curcuminoids using Captiva 96 wells plate EMR lipid.

[0372] EMR-Lipid protocol for the human plasma treatment before injection is as depicted below.

[0373] The LC/MS conditions were as follows. The autosampler (5° C.) and LC system was an Agilent Infinity II 1290 integrated system. Agilent 6420 Triple quadrupole mass spectrometer was used during the study, with electrospray ionization. The metabolites were eluted from the BEH C18 column (100×2.1 mm, 1.7 μm; Waters) with a mobile phase consisting of 0.1% formic acid in water in HPLC grade (solvent A) and 0.1% formic acid in acetonitrile (solvent B), at a flow rate of 0.4 mL/min and a column temperature of 35° C. The elution was in gradient from 40-60% B at 0-3 min. The injection volume was 2 μL for standard and samples. For each reference compound, a relevant transition of the precursor-to-product ions was detected with the utilization of the multiple reaction monitoring (MRM) mode. Each of the 5 analytes was determined in MS1 full scan tests and the product ions in MS/MS experiments. MRM transitions of each analyte was optimized using direct infusion and Optimizer B.08.00 workstation software solution (Agilent technologies, Santa Clara, Calif., USA). See Table 20 for the optimal selected conditions. The mass spectrometer parameters was set as follows: ESI source both in negative and positive mode; drying gas (N2) flow rate, 10 L/min; gas temperature, 350° C.; nebulizer, 40 psi; and capillary, 4.0 kV. The MS system was fully calibrated prior to running according to manufacturer's guidelines. Data analysis was carried out on Agilent MassHunter Quantitative/Qualitative analysis B.07.00 (Agilent technologies, Santa Clara, Calif., USA).

TABLE-US-00021 TABLE 20 Retention times (Tr), multiple reaction monitoring (MRM) transitions, and optimized tandem mass spectrometry (MS/MS) detection parameters of 5 curcuminoids and internal standard. Prec. Prod. Ion Q1 ions Q3 Dwell Mass Mass time Frag Cpd Name Rt (min) ISTD ? (Da) (Da) (ms) (V) CE (V) Polarity Curcumin d6 3.05 Yes 375.2 180.0 50 65 24 Positive 177.0 50 120 20 Positive Curcumin 3.05 No 369.1 145.0 50 120 36 Positive 117.0 50 120 48 Positive DMC d7 2.89 Yes 346.1 151.0 50 105 32 Positive DMC 2.89 No 339.1 177.0 50 105 20 Positive 147.0 50 105 28 Positive 91.1 50 105 60 Positive BDMC d8 2.74 Yes 317.1 151.0 50 80 24 Positive BDMC 2.74 No 309.1 147.0 50 95 24 Positive 119.0 50 95 40 Positive 91.1 50 95 60 Positive HHC d6 1.34 Yes 379.2 182.1 50 110 20 Negative HHC 1.34 No 373.1 193.1 50 100 12 Negative 179.1 50 100 16 Negative 121.0 50 100 56 Negative THC d6 2.84 Yes 377.1 135.1 50 105 60 Negative THC 2.84 No 371.1 235.1 50 120 12 Negative 193.1 50 120 20 Negative 135.0 50 120 56 Negative

[0374] Analysis Population [0375] ITT population: All subjects randomized in the study having consumed at least one dose of the products (n=30) [0376] PP population: Subjects included in the ITT population who completed the study presenting no major protocol deviations (n=30). The following subjects were excluded from the PP population: [0377] Subject SN01-040-V5 for all parameters [0378] SAFETY population: All subjects randomized in the study having consumed at least one dose of the products (n=30)

TABLE-US-00022 TABLE 21 Description of the study population at baseline showing mean and standard deviation. Included subjects Number of subjects (N = 30) Women/Men 16 (53.3)/14 (46.7) Age (years) 33.6 (6.79) Weight (kg) (V0) 64.5 (11.09) BMI (kg/m.sup.2) (V0) 22.1 (2.13)

[0379] Software Environment [0380] Statistical analyses were performed by Biofortis using SAS® software version 9.3 (SAS Institute Inc., Cary, N.C., USA). [0381] Significance level [0382] For all statistical tests (two-tailed), the 0.05 level of significance was used to justify a claim of a statistically significant effect. [0383] Methods of handling missing data for kinetics [0384] If more than 2 values or 2 consecutive values are missing in the kinetics, the AUC calculation cannot be performed and the kinetics were considered as missing in the statistical analyses (no missing data replacement was performed); [0385] If a data is missing at T-10 time-point, the AUC calculation cannot be performed and the kinetics were considered as missing in the statistical analyses (no missing data replacement was performed); [0386] If a value (except the baseline value and the value at the last time-point) is missing in the kinetics, it was replaced by the value obtained using the CopyMean method developed by Genolini (Genolini, 2013); [0387] If a value at the last time-point (T1440=T24 h) of the kinetics is missing, no missing data replacement was performed. [0388] In case of non-complete kinetics after missing data handling, the AUC cannot be calculated.

[0389] .fwdarw.This method was applied on ITT and PP populations.

[0390] Derived Variables [0391] Total curcuminoids=Curcumin+DMC+BDMC+THC+HHC+Curcumin glucuronide+DMC glucuronide+BDMC glucuronide+THC glucuronide+HHC glucuronide+Curcumin sulfate+DMC sulfate+BDMC sulfate+THC sulfate+HHC sulfate

[0392] If all of these 15 elements are missing, total curcuminoids cannot be calculated. If at least one of these 15 elements is quantified, total curcuminoids were calculated. [0393] Total Parent compounds=sum of curcumin+DMC+BDMC [0394] Total Parent compounds and their relative sulfate and glucuronide metabolites=Curcumin+Curcumin glucuronide+Curcumin sulfate+DMC+DMC glucuronide+DMC sulfate+BDMC+BDMC glucuronide+BDMC sulfate [0395] Curcumin and its relative sulfate and glucuronide metabolites=Curcumin+Curcumin glucuronide+Curcumin sulfate [0396] DMC and its relative sulfate and glucuronide metabolites=DMC+DMC glucuronide+DMC sulfate [0397] BDMC and its relative sulfate and glucuronide metabolites=BDMC+BDMC glucuronide+BDMC sulfate [0398] Curcumin and all its relative metabolites=Curcumin+Curcumin glucuronide+Curcumin sulfate+THC+THC glucuronide+THC sulfate+HHC+HHC glucuronide+HHC sulfate [0399] Relative bioavailability between 0 to 24 hours=Ratio of the dose-normalized AUC0-24 h for the different tested formulation to the dose-normalized AUC0-24 h obtained for the reference product (turmeric extract 95% curcuminoids) [0400] Relative bioavailability between 0 to 8 hours=Ratio of the dose-normalized AUC0-8 h for the different tested formulation to the dose-normalized AUC0-8 h obtained for the reference product (turmeric extract 95% curcuminoids) [0401] Relative bioavailability between 0 to infinity=Ratio of the dose-normalized AUC0-∞ for the different tested formulation to the dose-normalized AUC0-∞ obtained for the reference product (turmeric extract 95% curcuminoids)

[0402] Data Handling of Values Below the Limit of Detection (LOD) [0403] Some values below the limit of detection (LOD) were identified for curcumin (native, glucuronide and sulfate) expressed as <<0.62>> in the database.

[0404] .fwdarw.The number and percentage of values under the LOD were given for each parameter and visit.

[0405] Production of Graphical Representations [0406] Quantitative variable on observed means: Box-and-whiskers plots for AUC parameters (illustrated in Figure below)

[0407] Checking of Statistical Tests Assumptions [0408] Assumptions of normality and homoscedasticity were investigated by graphical representations of residuals produced by statistical models. In case of strong deviation from normality and/or homoscedasticity, log transformation (log 10) of study endpoints was considered.

[0409] Note for production of results: [0410] STE=Standard turmeric powder extract 95% curcuminoids 1500 mg [0411] TEP=Curcumin C3 complex California Gold Nutrition (1500 mg C3 Complex®) [0412] NOV=Curcumin Cell'Innov (1000 mg Novasol®) [0413] PHYT=Curcuma Platinum MannaVital (1000 mg Meriva®) [0414] Turmipure GOLD™=Turmipure Gold 30% curcuminoids 300 mg

[0415] Statistical Methodology [0416] Primary endpoint: Dose-normalized AUC between 0 and 24 hours was analyzed using the following mixed model for repeated measurements (SAS® PROC MIXED, statistical model n° 1):

Y=Product+Visit+Baseline+Subject.sub.random

[0417] with: [0418] Y: Dose-normalized AUC between 0 and 24 hours of analyt plasmatic concentration; [0419] Product: Turmipure Gold™, STE, TEP, NOV, PHYT; [0420] Visit: Visit V1 to V5; [0421] Baseline: Parameter's value at T-10 time-point (T0 for AUC calculation); [0422] Subject.sub.random: Random factor. [0423] If significant Visit effect (p<0.05): secondary analysis realized on first period (visit) in order to assess the product effect. [0424] Comparison between products of interest.fwdarw.Turmipure Gold™ compared to STE

[0425] Additional Analysis: Investigation of Gender Effect [0426] Gender effect was investigated in this study using the following mixed model for repeated measurements (SAS® PROC MIXED, statistical model n° 2):

Y=Product+Visit+Gender+Product*Gender+Baseline+Subject.sub.random

[0427] With: [0428] Y: Endpoint; [0429] Product: Turmipure Gold™, STE, TEP, NOV, PHYT; [0430] Visit: Visit V1 to V5; [0431] Gender: Female or Male; [0432] Product*Gender: interaction between the product and the gender; [0433] Baseline: Parameter's value at T-10 time-point (T0 for AUC calculation); [0434] Subject.sub.random: Random factor. [0435] Comparisons between products of interest; [0436] Turmipure GOLD™ compared to STE; [0437] TEP compared to STE; [0438] NOV compared to STE; [0439] PHYT compared to STE; [0440] TEP compared to Turmipure GOLD™; [0441] NOV compared to Turmipure GOLD™; [0442] PHYT compared to Turmipure GOLD™ [0443] If significant Visit effect (p<0.05): secondary analysis realized on first period (visit) in order to assess the product effect. [0444] If significant Product*Gender interaction effect (p<0.05): treatment effect investigated in men and women separately (with production of descriptive statistics and graphic representations) [0445] If significant Product*Gender interaction effect (p>0.05): treatment effect investigated globally (women and men together)

[0446] Statistical Methodology for Secondary Endpoints (Except for Relative Bioavailability) [0447] Secondary endpoints were analyzed using the following mixed model for repeated measurements (SAS® PROC MIXED, statistical model n° 1):

Y=Product+Visit+Baseline+Subject.sub.random [0448] If significant Visit effect (p<0.05): secondary analysis realized on first period (visit) in order to assess the product effect. [0449] Comparison between products of interest: [0450] Turmipure GOLD™ compared to STE; [0451] TEP compared to STE; [0452] NOV compared to STE; [0453] PHYT compared to STE; [0454] TEP compared to Turmipure GOLD™; [0455] NOV compared to Turmipure GOLD™; [0456] PHYT compared to Turmipure GOLD™ [0457] Statistical methodology for relative bioavailability [0458] Secondary endpoints were analyzed using the following mixed model for repeated measurements (SAS® PROC MIXED, statistical model n° 1):

Y=Product+Visit+Subject.sub.random [0459] If significant Visit effect (p<0.05): secondary analysis realized on first period (visit) in order to assess the product effect. [0460] Comparison between products of interest: [0461] TEP compared to Turmipure GOLD™; [0462] NOV compared to Turmipure GOLD™; [0463] PHYT compared to Turmipure GOLD™

[0464] Summary of the Results of the Primary Endpoints

[0465] Table 22 and 23: Initial Analysis of ITT and PP Populations

TABLE-US-00023 ITT population Between-group analysis Product effect- Comparison between Statistical Endpoint Statistical significance Products significance Dose-normalized of Turmipure GOLD ™ vs STE Turmipure > STE AUC.sub.0-24h of Total (primary endpoint) curcuminoids TEP vs STE (log-transformed data) NOV vs STE NOV > STE PHYT vs STE PHYT > STE TEP vs Turmipure GOLD ™ TEP < Turmipure NOV vs Turmipure GOLD ™ NOV > Turmipure PHYT vs Turmipure GOLD ™ PHYT < Turmipure

TABLE-US-00024 PP population Between-group analysis Product effect- Comparison between Statistical Endpoint Statistical significance Products significance Dose-normalized of Turmipure GOLD ™ vs STE Turmipure > STE AUC.sub.0-24h of Total (primary endpoint) curcuminoids TEP vs STE (log-transformed data) NOV vs STE NOV > STE PHYT vs STE PHYT > STE TEP vs Turmipure GOLD ™ TEP < Turmipure NOV vs Turmipure GOLD ™ NOV > Turmipure PHYT vs Turmipure GOLD ™ PHYT < Turmipure

[0466] Table 24 and 25: Additional Analysis of ITT and PP Populations (Investigation of Gender Effect)

TABLE-US-00025 ITT population Between-group analysis Product*Gender effect- Comparison between Statistical Endpoint Statistical significance Products significance Dose-normalized of Turmipure GOLD ™ vs STE Turmipure > STE AUC.sub.0-24h of Total (primary endpoint) curcuminoids TEP vs STE (log-transformed data) NOV vs STE NOV > STE PHYT vs STE PHYT > STE TEP vs Turmipure GOLD ™ TEP <Turmipure NOV vs Turmipure GOLD ™ NOV >Turmipure PHYT vs Turmipure GOLD ™ PHYT <Turmipure

TABLE-US-00026 PP population Between-group analysis Product*Gender effect- Comparison between Statistical Endpoint Statistical significance Products significance Dose-normalized of Turmipure GOLD ™ vs STE Turmipure > STE AUC.sub.0-24h of Total (primary endpoint) curcuminoids TEP vs STE (log-transformed data) NOV vs STE NOV > STE PHYT vs STE PHYT > STE TEP vs Turmipure GOLD ™ TEP <Turmipure NOV vs Turmipure GOLD ™ NOV >Turmipure PHYT vs Turmipure GOLD ™ PHYT <Turmipure p-value < 0.05 (statistically signifcant); p-value > 0.05 (statistically non-significant

TABLE-US-00027 TABLE 26 Dose-normalized of AUC.sub.0-24h of Total curcuminoids Turmipure STE product TEP productNOV productPHYT product GOLD ™ product Variable Statistics (n = 30) (n = 30) (n = 30) (n = 30) (n = 30) Dose-normalized N 30 30 30 29 30 AUC.sub.0-24H of Total N Miss 0 0 0 1 0 curcuminoids Mean (SD) 3.7 (1.75) 3.2 (1.69) 136.1 (37.40) 13.0 (9.65) 72.9 (25.49) (ng .Math. h/mL/mg) (Min; Max) (0.7; 9.7) (0.8; 8.7) (69.8; 220.8) (1.7; 42.0) (16.4; 139.6) Median (Q1; Q3) 3.7 (2.5; 4.3) 3.0 (2.1; 4.1) 141.8 (105.6; 10.7 (6.2; 17.0) 69.7 (55.2; 87.8) 157.8)

[0467] From the above analysis, it was concluded that for the ITT population:

[0468] Between-Group Analysis (all Genders Taken Together) [0469] No significant visit is identified (p=0.2245).fwdarw.Consequently, the analysis was performed on all visits. [0470] Significant Product effect (p<0.0001): [0471] Primary endpoint: There is a statistically significant difference between Turmipure GOLD™ and STE (adjusted p<0.0001; diff[adjusted CI95%]=1.32 [1.18; 1.46]). [0472] Others comparisons: [0473] TEP vs STE (adjusted p=0.6948) [0474] NOV vs STE (adjusted p<0.0001; diff[adjusted CI95%]=1.62 [1.48; 1.76]).fwdarw.NOV>STE [0475] PHYT vs STE (adjusted p<0.0001; diff[adjusted CI95%]=0.48 [0.34; 0.62]).fwdarw.PHYT>STE [0476] TEP vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=−1.39 [−1.53;−1.25]).fwdarw.TEP<Turmipure GOLD™ [0477] NOV vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=0.29 [0.15; 0.43]).fwdarw.NOV>Turmipure GOLD™ [0478] PHYT vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=−0.84 [−0.99;−0.70]).fwdarw.PHYT<Turmipure GOLD™ [0479] Additional analysis: Investigation of Gender effect [0480] No significant Visit (p=0.2456) and Product*Gender interaction (p=0.3804) effects: Analysis performed all gender taken together and on all visits. [0481] Significant Product effect (p<0.0001): [0482] TEP vs STE (adjusted p=0.7091) [0483] NOV vs STE (adjusted p<0.0001; diff[adjusted CI95%]=1.61 [1.47; 1.75]).fwdarw.NOV>STE [0484] PHYT vs STE (adjusted p<0.0001; diff[adjusted CI95%]=0.48 [0.34; 0.62]).fwdarw.PHYT>STE [0485] Turmipure GOLD™ vS STE (adjusted p<0.0001; diff[adjusted CI95%]=1.32 [1.18; 1.46])+Turmipure GOLD™>STE [0486] TEP vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=−1.39 [−1.52;−1.25]).fwdarw.TEP<Turmipure GOLD™ [0487] NOV vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=0.29 [0.15; 0.43]).fwdarw.NOV>Turmipure GOLD™ [0488] PHYT vs Turmipure GOLD™ (adjusted p<0.0001; diff[adjusted CI95%]=−0.84 [−0.98;−0.70]).fwdarw.PHYT<Turmipure GOLD™

[0489] For the PP population: [0490] Between-group analysis (all genders taken together) [0491] Results are similar to results observed on ITT population. [0492] Additional analysis: Investigation of Gender effect [0493] Results are similar to results observed on ITT population.

[0494] Summary of the Results of the Secondary Endpoints

[0495] The results of the secondary endpoints are shown in the tables below for both the ITT (Tables 27 to 43) and PP (Tables 44 to 60) Populations respectively. The graphical representation of the data is provided in FIGS. 24 to 37.

[0496] The following key applies to each table, indicating the statistical significance of each result.

TABLE-US-00028 p-value < 0.05 (statistically significant); p-value ≥ 0.10 (statistically non significant); *Analysis performed on V1 visit only (n = 30)

TABLE-US-00029 TABLE 27 Analysis of total curcuminoids (ng/mL) between products in the ITT population. Between-group analysis-Satisfied significance of comparisons between products (adjusted p-value, Tokey adjustment) Termipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24h (ng .Math. h/mL/mg) [Log10] Turmipure > STE STE STE Turmipure Turmipure Turmipure AUC0-24h [ng .Math. h/mL] [Log10] STE STE Turmipure Turmipure Turmipure DN of AUC0-8h Turmipure > STE (ng .Math. h/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8h (ng .Math. h/mL) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] DN of AUCa-infinity (ng .Math. h/mL/mg) Turmipure > STE STE Turmipure Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) [Log10] Normalized Cmax (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax (ng .Math. h/mL) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] Ret. bio. between 0 and 24 h Turmipure Turmipure Turmipure [Log10] Ret. bio. between 0 and 8 h Turmipure Turmipure Turmipure [Log10] Ret. bio. between 0 and infinity [Log10] Half-life (minutes) [Log10] Turmipure > STE STE Turmipure Turmipure Terminal elimination rate constant Turmipure > STE STE Turmipure Turmipure [Log10] Tmax minutes [Log10] STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00030 TABLE 28 Analysis of total curcumin (ng/mL) between products in the ITT population. Between-group analysis-Satisfied significance of comparisons between products (adjusted p-value, Tokey adjustment) Termipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24h (ng .Math. h/mL/mg) [Log10] Turmipure > STE Turmipure AUC0-24h [ng .Math. h/mL] [Log10] DN of AUC0-8h (ng .Math. h/mL/mg) [Log10] AUC0-8h (ng .Math. h/mL) [Log10] DN of AUCa-infinity (ng .Math. h/mL/mg) Turmipure > STE Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) [Log10] Normalized Cmax (ng .Math. h/mL/mg) [Log10] Cmax (ng .Math. h/mL) [Log10] Ret. bio. between 0 and 24 h [Log10] Ret. bio. between 0 and 8 h Turmipure [Log10] Ret. bio. between 0 and infinity [Log10] Half-life (minutes) [Log10] Terminal elimination rate constant [Log10] Tmax minutes [Log10] indicates data missing or illegible when filed

TABLE-US-00031 TABLE 29 Analysis of total curcumin (ng/mL) between products in the ITT population. Between-group analysis-Satisfied significance of comparisons between products (adjusted p-value, Tokey adjustment) Termipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24h (ng .Math. h/mL/mg) [Log10] Turmipure > STE STE STE Turmipure Turmipure Turmipure AUC0-24h [ng .Math. h/mL] [Log10] STE STE Turmipure Turmipure DN of AUC0-8h (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8h (ng .Math. h/mL) STE STE Turmipure Turmipure [Log10] DN of AUCa-infinity (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) Turmipure [Log10] Normalized Cmax (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax (ng .Math. h/mL) STE STE Turmipure Turmipure [Log10] Ret. bio. between 0 and 24 h Turmipure Turmipure Turmipure [Log10] Ret. bio. between 0 and 8 h Turmipure Turmipure Turmipure [Log10] Ret. bio. between 0 and infinity Turmipure Turmipure [Log10] Half-life (minutes) [Log10] Turmipure < STE STE Terminal elimination rate constant Turmipure > STE STE [Log10] Tmax minutes [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00032 TABLE 30 Analysis of curcumin sulfate (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Visit Product Turmipure TEP vs Turmipure NOV vs Turmipure PHYT vs Turmipure Endpoint effect effect GOLD ™ vs STE TEP vs STE NOV vs STE PHYT vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h p = 0.0037* p < 0.0001 p < 0.0001;     p < 0.0001; NOV > p < 0.0001; PHYT > p < 0.0001; TEP <     p = 0.0017; PHYT < (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure [Log10] AUC0-24 h p = 0.0037*                                 (ng .Math. h/mL) [Log10] DN of AUC0-8 h p = 0.0   * p < 0.0001 p < 0.0001;     p < 0.0001; NOV > p < 0.0001; PHYT > p < 0.0001; TEP < p = 0.0041; NOV > p < 0.0001; PHYT < (ng .Math. h/mL/mg) Turmipure > STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h (ng .Math. h/mL) [Log10] p = 0.0293* p = 0.00                                DN of AUC0-infinity     p < 0.0001 p < 0.0001;     p < 0.0001; NOV > p < 0.0001; PHYT > p < 0.0001; TEP <     p < 0.0001; PHYT < (ng .Math. h/mL/mg) [Log10] Turmipure > STE STE STE Turmipure Turmipure AUC0-infinity                                     (ng .Math. h/mL) [Log10] Normalized Cmax p = 0.0003* p < 0.0001 p < 0.0001;     p < 0.0001; NOV > p < 0.0001; PHYT > p < 0.0001; TEP < p < 0.0001; NOV > p = 0.0005; PHYT < (ng/mL/mg) [Log10] Turmipure > STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL) p = 0.0003* p < 0.0001         p = 0.0   ; NOV >         p = 0.002   ; NOV >     [Log10] STE Turmipure Rel. bio. between 0 and 24 h p = 0.0260* p < 0.0001                 p < 0.0001; TEP <     p = 0.0015; PHYT < [Log10] Turmipure Turmipure Rel. bio. between 0 and 8 h     p < 0.0001                 p < 0.0001; TEP < p = 0.0002; NOV > p < 0.0001; PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between 0 and p = 0.0100* p = 0.0143                             infinity [Log10] Half-life (minutes)                                     [Log10] Terminal elimination                                     rate constant [Log10] Tmax (minutes)     p < 0.0001         p < 0.0001; NOV <     p = 0.0   95; TEP > p < 0.0001, NOV < p = 0.0043; PHYT > [Log10] STE Turmipure Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00033 TABLE 31 Analysis of curcumin and its relative sulfate and glucuronide metabolites (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE PHYT vs STE GOLD ™ GOLD ™ GOLD ™ DN of     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; PHYT < AUC0-24 h 0.0001 Turmipure > NOV > STE PHYT > STE TEP < NOV > Turmipure (ng .Math. h/mL/mg) STE Turmipure Turmipure [Log10] AUC0-24 h     p <         p = 0.0173; p < 0.0001;     p = 0.0031; p < 0.0001; PHYT < (ng .Math. h/mL) 0.0001 NOV > STE PHYT > STE NOV > Turmipure [Log10] Turmipure DN of AUC0-8 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; PHYT < (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > STE PHYT > STE TEP < NOV > Turmipure [Log10] STE Turmipure Turmipure AUC0-8 h     p <         p < 0.0001; p < 0.0001; p < 0.0265; p < 0.0001; p < 0.0001; PHYT < (ng .Math. h/mL) 0.0001 NOV > STE PHYT < STE TEP < NOV > Turmipure [Log10] Turmipure Turmipure DN of AUC0- p = p < p < 0.0001;     p < 0.0001; p = 0.0158; p < 0.0001;     p = 0.0333; PHYT < infinity 0.0033* 0.0001 Turmipure > NOV > STE PHYT > STE TEP < Turmipure (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-infinity p =                                 (ng .Math. h/mL) 0.0033* [Log10] Normalized p = p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0018; PHYT < Cmax 0.040   * 0.0001 Turmipure > NOV > STE PHYT > STE TEP < NOV > Turmipure (ng/mL/mg) STE Turmipure Turmipure [Log10] Cmax (ng/mL) p = p <         p < 0.0001;         p < 0.0001;     [Log10] 0.040   * 0.0001 NOV > STE NOV > Turmipure Rel. bio.     p <                 p < 0.0001; p < 0.0001; p = 0.0001; between 0 and 0.0001 TEP < NOV > PHYT < 24 h [Log10] Turmipure Turmipure Turmipure Rel. bio. between     p <                 p < 0.0001; p < 0.0001; p < 0.0001; PHYT < 0 and 8 h 0.0001 TEP < NOV > Turmipure [Log10] Turmipure Turmipure Rel. bio. between p = p =                 p = 0.0190;         0 and infinity 0.00   * 0.0002 TEP < [Log10] Turmipure Half-life     p =                         p = 0.0062; PHYT > (minutes) 0.0010 Turmipure [Log10] Terminal     p =                         p = 0.0062; PHYT < elimination 0.0010 Turmipure rate constant [Log10] Tmax (minutes)     p <         p < 0.0001;         p < 0.0001; p = 0.041   ; [Log10] 0.0001 NOV < STE NOV < PHYT > Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00034 TABLE 32 Analysis of DMC glucuronide (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001;     (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > [Log10] STE STE STE Turmipure Turmipure AUC0-24 h     p < 0.0001         p = 0.0010;         p < 0.0001;     (ng .Math. h/mL) NOV > NOV > [Log10] STE Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0131; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001         p < 0.0001;         p < 0.0001;     (ng .Math. h/mL) NOV > NOV > [Log10] STE Turmipure DN of AUC0-infinity     p < 0.0001 p < 0.0001;     p < 0.0001; p = 0.0135; p < 0.0001; p = 0.0187;     (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > [Log10] STE STE STE Turmipure Turmipure AUC0-infinity                                     (ng .Math. h/mL) [Log10] Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p = 0.0066; p < 0.0001; p < 0.0001; p = 0.0   ; (ng/mL/mg) [Log10] Turmipure > NOV > PHYT > TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001         p < 0.0001;         p < 0.0001;     [Log10] NOV > NOV > STE Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p < 0.0001;     between TEP < NOV > 0 and 24 h Turmipure Turmipure [Log10] Rel. bio.     p < 0.0001                 p < 0.0001; p < 0.0001; p = 0.0207; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel. bio. between 0     p < 0.0001                 p < 0.0001;         and infinity [Log10] TEP < Turmipure Half-life (minutes)     p = 0.0035                             [Log10] Terminal elimination     p = 0.0002         p = 0.0008;         p = 0.02   2;     rate constant NOV > NOV > [Log10] STE Turmipure Tmax (minutes)     p = 0.0108                             [Log10] indicates data missing or illegible when filed

TABLE-US-00035 TABLE 33 Analysis of DMC sulfate (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure vs Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001;     p = 0.0003; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > PHYT > TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure AUC0-24 h     p =         p = 0.00                    (ng .Math. h/mL) 0.0051 NOV < [Log10] STE DN of AUC0-8 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > PHYT > TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure AUC0-8 h                                     (ng .Math. h/mL) [Log10] DN of     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001;     p = 0.0101; AUC0-infinity 0.0001 Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity     p =         p = 0.0019;                 (ng .Math. h/mL) 0.0016 NOV < [Log10] STE Normalized     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0106; p < 0.0001; Cmax (ng/mL/ 0.0001 Turmipure > NOV > PHYT > TEP < NOV > PHYT < mg) [Log10] STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)                                 [Log10] Rel. bio. between     p <                 p < 0.0001;     p = 0.0007; 0 and 24 h 0.0001 TEP < PHYT < [Log10] Turmipure Turmipure Rel. bio. between     p <                 p < 0.0001;     p < 0.0001; 0 and 8 h 0.0001 TEP < PHYT < [Log10] Turmipure Turmipure Rel. bio. between     p <                 p < 0.0001;         0 and infinity 0.0001 TEP < [Log10] Turmipure Half-life (minutes)                                     [Log10] Terminal                                     elimination rate constant [Log10] Tmax (minutes)     p <         p < 0.0001;         p = 0.0022;     [Log10] 0.0001 NOV < NOV < STE Turmipure indicates data missing or illegible when filed

TABLE-US-00036 TABLE 34 Analysis of DMC and its relative sulfate and glucuronide metabolites (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0293; p = 0.0005; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h                                     (ng .Math. h/mL) [Log10] DN of AUC0-8 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p =         p = 0.0112;         p = 0.0075;     (ng .Math. h/mL) 0.00    NOV > NOV > [Log10] STE Turmipure DN of     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; AUC0-infinity 0.0001 Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity     p =                             (ng .Math. h/mL) 0.0150 [Log10] Normalized     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p = 0.00   ; Cmax (ng/mL/ 0.0001 Turmipure > NOV > PHYT > TEP < NOV > PHYT < mg) [Log10] STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p <         p < 0.0001;         p < 0.0001;     [Log10] 0.0001 NOV > NOV > STE Turmipure Rel. bio.     p <                 p < 0.0001;     p =    .0052; between 0.0001 TEP < PHYT < 0 and 24 h Turmipure Turmipure [Log10] Rel. bio.     p <                 p < 0.0001; p = 0.0004; p < 0.0001; between 0 0.0001 TEP < NOV > PHYT < and 8 h Turmipure Turmipure Turmipure [Log10] Rel. bio.     p <                 p < 0.0001;     p = 0.0006; between 0 and 0.0001 TEP < PHYT < infinity [Log10] Turmipure Turmipure Half-life     p =         p = 0.0019;         p = 0.0018;     (minutes) 0.0004 NOV < NOV < [Log10] STE Turmipure Terminal     p =         p = 0.0019;         p = 0.0018;     elimination 0.0004 NOV > NOV > rate constant STE Turmipure [Log10] Tmax (minutes)     p <         p    0001; p = 0.0117;     p = 0.0060; p = 0.0003; [Log10] 0.0001 NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00037 TABLE 35 Analysis of BDMC glucuronide (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p <         p < 0.0001;         p = 0.0001;     (ng .Math. h/mL/mg) 0.0001 NOV > NOV > [Log10] STE Turmipure AUC0-24 h     p = p = 0.0003;                 p = 0.0040;     (ng .Math. h/mL) 0.0003 Turmipure < NOV > [Log10] STE Turmipure DN of AUC0-8 h     p <         p < 0.0001;         p < 0.0001;     (ng .Math. h/mL/mg) 0.0001 NOV > NOV > [Log10] STE Turmipure AUC0-8 h     p < p < 0.0003; p = 0.0290;     p = 0.000   ; p < 0.0153; p < 0.0001;     (ng .Math. h/mL) 0.0001 Turmipure < TEP < PHYT < TEP > NOV > [Log10] STE STE STE Turmipure Turmipure DN of AUC0-infinity     p <         p = 0.000   ;         p = 0.0010;     (ng .Math. h/mL/mg) 0.0001 NOV > NOV > [Log10] STE Turmipure AUC0-infinity     p = p = 0.0060;         p = 0.0263; p = 0.02   2; p = 0.0439;     (ng .Math. h/mL) 0.0009 Turmipure < PHYT < TEP > NOV > [Log10] STE STE Turmipure Turmipure Normalized Cmax     p <         p < 0.0001;     p = 0.0007; p < 0.0001;     (ng/mL/mg) [Log10] 0.0001 NOV > TEP < NOV > STE Turmipure Turmipure Cmax (ng/mL)     p < p < 0.000   ; p = 0.00   ;     p < 0.0001;     p < 0.0001;     [Log10] 0.0001 Turmipure < TEP < PHYT < NOV > STE STE STE Turmipure Rel. bio.     p <                 p = 0.0291; p = 0.0005;     between 0 and 24 h 0.0001 TEP < NOV > [Log10] Turmipure Turmipure Rel. bio.     p <                     p < 0.0001;     between 0 and 8 h 0.0001 NOV > [Log10] Turmipure Rel. bio. between 0     p =                             and infinity [Log10] 0.0275 Half-life (minutes)     p < p = 0.0020;         p = 0.004   ; p = 0.0116; p = 0.0075;     [Log10] 0.0001 Turmipure < PHYT < TEP > NOV > STE STE Turmipure Turmipure Terminal elimination     p < p = 0.006   ; p = 0.0252;     p = 0.0015;     p < 0.0001;     rate constant 0.0001 Turmipure < TEP < PHYT < NOV > [Log10] STE STE STE Turmipure Tmax (minutes)     p =                     p = 0.01   ;     [Log10] 0.0275 NOV > Turmipure indicates data missing or illegible when filed

TABLE-US-00038 TABLE 36 Analysis of BDMC sulfate (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p = 0.0055                 p = 0.023   ;         (ng .Math. h/mL/mg) TEP < [Log10] Turmipure AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0008; p < 0.0001;     p = 0.0280; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > PHYT > [Log10] STE STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001         p = 0.0101;     p < 0.0001;         (ng .Math. h/mL/mg) NOV > TEP < [Log10] STE Turmipure AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001;         (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > [Log10] STE STE STE Turmipure DN of AUC0-infinity                                     (ng .Math. h/mL/mg) [Log10] AUC0-infinity     p < 0.0001 p < 0.0001;     p = 0.0002; p = 0.0059; p < 0.0001;         (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > [Log10] STE STE STE Turmipure Normalized Cmax p = 0.0424* p = 0.0248                             (ng/mL/mg) [Log10] Cmax (ng/mL) p = 0.0300* p = 0.0029 p = 0.0109;     p = 0.0   2; p = 0.0109;             [Log10] Turmipure < NOV < PHYT < STE STE STE Rel. bio. between     p = 0.0320                             0 and 24 h [Log10] Rel. bio. between     p = 0.0002                 p = 0.0024;         0 and 8 h TEP < [Log10] Turmipure Rel. bio. between 0                                     and infinity [Log10] Half-life (minutes)     p < 0.0001 p < 0.0001;         p = 0.04   6; p < 0.00   1;         [Log10] Turmipure < PHYT < TEP > STE STE Turmipure Terminal elimination     p = 0.0012         p = 0.0021;                 rate constant NOV > [Log10] STE Tmax (minutes)     p = 0.0219                         p = 0.0367; [Log10] PHYT > Turmipure indicates data missing or illegible when filed

TABLE-US-00039 TABLE 37 Analysis of BDMC and its relative sulfate and glucuronide metabolites (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001         p = 0.00   0;     p = 0.0015;         (ng .Math. h/mL/mg) NOV > TEP < [Log10] STE Turmipure AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p = 0.0004; p < 0.0001;     p = 0.0111; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > PHYT > [Log10] STE STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0   01;     p < 0.0001;     p < 0.0001; p = 0.0452;     (ng .Math. h/mL/mg) Turmipure > NOV > TEP < NOV > [Log10] STE STE Turmipure Turmipure AUC0-8 h     p < p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0064;     (ng .Math. h/mL) 0.000    Turmipure < NOV < PHYT < TEP > NOV > [Log10] STE STE STE Turmipure Turmipure DN of AUC0-infinity     p = 0.0033             p = 0.0350;             (ng .Math. h/mL/mg) PHYT > [Log10] STE AUC0-infinity     p < 0.0001 p < 0.0001;     p = 0.0003;     p < 0.0001;     p = 0.01   8; (ng .Math. h/mL) Turmipure < NOV < TEP > PHYT > [Log10] STE STE Turmipure Turmipure Normalized Cmax     p < 0.0001 p = 0.001   ;     p < 0.0001;     p < 0.0001; p < 0.0001; p = 0.0054; (ng/mL/mg) [Log10] Turmipure > NOV > TEP < NOV > PHYT < STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001 p < 0.0001; p = 0.0010; p = 0.0086; p < 0.0001; p < 0.0001; p < 0.0001;     [Log10] Turmipure < TEP < NOV < PHYT < TEP > NOV > STE STE STE STE Turmipure Turmipure Rel. bio. between     p = 0.0004                 p = 0.0303;         0 and 24 h TEP < [Log10] Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001;         0 and 8 h TEP < [Log10] Turmipure Rel. bio. between 0     p =                             and infinity [Log10] 0.04   3 Half-life (minutes)     p < 0.0001 p = 0.0025;     p = 0.0201;     p < 0.0001;     p = 0.0310; [Log10] Turmipure < NOV < TEP > PHYT > STE STE Turmipure Turmipure Terminal elimination     p < 0.0001         p < 0.0001;         p = 0.0003;     rate constant NOV > NOV > [Log10] STE Turmipure Tmax (minutes)     p =                         p = 0.0   07; [Log10] 0   0133 PHYT > Turmipure indicates data missing or illegible when filed

TABLE-US-00040 TABLE 38 Analysis of THC glucuronide (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < p < 0.0001;     p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > TEP < NOV > PHYT < [Log10] STE STE Turmipure Turmipure Turmipure AUC0-24 h     p <         p = 0.0212; p < 0.0001; p = 0.028   ;     p < 0.0001; (ng .Math. h/mL) 0.0001 NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure DN of AUC0-8 h     p < p < 0.0001;     p < 0.0001;     p     0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > TEP < NOV > PHYT < [Log10] STE STE Turmipure Turmipure Turmipure AUC0-8 h     p <         p = 0.000   ; p < 0.0001; p = 0.0020;     p < 0.0001; (ng .Math. h/mL) 0.0001 NOV > STE PHYT < TEP < PHYT < [Log10] STE Turmipure Turmipure DN of AUC0-infinity     p < p < 0.0001;     p < 0.0001; p = 0.0387; p < 0.0001;     p < 0.0001; (ng .Math. h/mL/mg) 0.0001 Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure AUC0-infinity     p <             p < 0.0001; p = 0.0171;     p < 0.0001; (ng .Math. h/mL) 0.0001 PHYT < TEP < PHYT < [Log10] STE Turmipure Turmipure Normalized Cmax     p < p < 0.0001;     p < 0.0001; p = 0.0173; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/ 0.0001 Turmipure > NOV > PHYT > TEP < NOV > PHYT < mg) [Log10] STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p <         p < 0.0001; p < 0.0001; p = 0.0359;     p < 0.0001; [Log10] 0.0001 NOV > PHYT < TEP < PHYT < STE STE Turmipure Turmipure Rel. bio.     p <                 p < 0.0001; p = 0.0030; p < 0.0001; between 0 0.0001 TEP < NOV > PHYT < and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio.     p <                 p < 0.0001; p < 0.0001; p < 0.0001; between 0 0.0001 TEP < NOV > PHYT < and 8 h Turmipure Turmipure Turmipure [Log10] Rel. bio.     p <                 p < 0.0001;     p < 0.0001; between 0 and 0.0001 TEP < PHYT < infinity [Log10] Turmipure Turmipure Half-life (minutes)     p <             p < 0   0001;         p < 0.0001; [Log10] 0.0001 PHYT < STE PHYT < Turmipure Terminal elimination     p <                             rate constant 0.0001 [Log10] Tmax (minutes)     p <             p = 0.0004;         p = 0.0003; [Log10] 0.0001 PHYT < STE PHYT < Turmipure indicates data missing or illegible when filed

TABLE-US-00041 TABLE 39 Analysis of THC sulfate (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h p =                                 (ng .Math. h/mL/mg) 0.0057* [Log10] AUC0-24 h p =                                 (ng .Math. h/mL) 0.0328* [Log10] DN of AUC0-8 h p =                                 (ng .Math. h/mL/mg) 0.0109* [Log10] AUC0-8 h     p < 0.0001             p = 0.0067;         p = 0.0003; (ng .Math. h/mL) PHYT < PHYT < [Log10] STE Turmipure DN of AUC0-infinity     p = 0.0013         p = 0.0327;                 (ng .Math. h/mL/mg) NOV > [Log10] STE AUC0-infinity     p = 0.0134                             (ng .Math. h/mL) [Log10] Normalized Cmax p = p = 0.0438                             (ng/mL/mg) [Log10] 0.0007* Cmax (ng/mL)     p = 0.0006             p = 0.0315;         p = 0.0005; [Log10] PHYT < PHYT < STE Turmipure Rel. bio.     p = 0.0022                             between 0 and 24 h [Log10] Rel. bio.     p < 0.0001                 p = 0   0038;     p = 0.01   8; between 0 and 8 h TEP < PHYT < [Log10] Turmipure Turmipure Rel. bio. between 0                                     and infinity [Log10] Half-life (minutes)     p = 0.0134                             [Log10] Terminal elimination     p = 0.0412                         p = 0.0184; rate constant PHYT < [Log10] Turmipure Tmax (minutes) p =                                 [Log10] 0.0309* indicates data missing or illegible when filed

TABLE-US-00042 TABLE 40 Analysis of HHC glucuronide (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0013; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h     p < 0.0001 p = 0.0002;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > TEP < NOV > PHYT < [Log10] STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001 p = 0.0015;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of AUC0-infinity     p < 0.0001 p < 0.0001;     p < 0.0001; p = 0.0352; p < 0.0001; p = 0.023   ; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-infinity     p < 0.0001         p = 0.0133; p < 0.0001; p = 0.0192;     p < 0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/mg) [Log10] Turmipure > NOV > PHYT > TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001 p = 0.0002;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; [Log10] Turmipure > NOV > PHYT < TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p = 0.0   02; p < 0.0001; between 0 and 24 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; between 0 and 8 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between 0     p < 0.0001                 p < 0.0001;     p < 0.0001; and infinity [Log10] TEP < PHYT < Turmipure Turmipure Half-life (minutes)                                     [Log10] Terminal elimination                                     rate constant [Log10] Tmax (minutes)     p < 0.0001         p < 0.0001; p = 0.03   ;     p < 0.0001; p < 0.0001; [Log10] NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00043 TABLE 41 Analysis of HHC sulfate (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h     p < 0.0001 p = 0.0010;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0   02; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001 p = 0.0002;     p < 0.0001; p < 0.0001; p < 0.0001;     p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of AUC0-infinity     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.000   ; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-infinity     p < 0.0001         p = 0.0313; p < 0.0001; p = 0.0228;     p < 0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/mg) [Log10] Turmipure > NOV > PHYT > TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; [Log10] Turmipure > NOV > PHYT < TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; between 0 and 24 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p = 0.0   08; p < 0.0001; between 0 and 8 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between 0 and     p < 0.0001                 p < 0.0001; p = 0.0009; p < 0.0001; infinity [Log10] TEP < NOV > PHYT < Turmipure Turmipure Turmipure Half-life (minutes)     p < 0.0001 p = 0.0011;     p = 0.0011;     p = 0.0038;         [Log10] Turmipure < NOV < TEP > STE STE Turmipure Terminal elimination     p < 0.0001 p = 0.0011;     p = 0.0011;     p = 0.0038;         rate constant Turmipure > NOV > TEP < [Log10] STE STE Turmipure Tmax (minutes)     p < 0.0001         p < 0.0001; p < 0.0001;     p < 0.000   ; p < 0.0001; [Log10] NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00044 TABLE 42 Analysis of curcumin and all its relative metabolites (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h     p < 0.0001 p = 0.0424;     p < 0.0001; p < 0.0001; p = 0.0005;     p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001 p = 0.0002;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0073; p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure DN of AUC0-infinity p = 0.0328* p < 0.0001 p = 0.0003;     p < 0.0001;     p < 0.0001;     p = 0.0054; (ng .Math. h/mL/mg) Turmipure > NOV > TEP < PHYT < [Log10] STE STE Turmipure Turmipure AUC0-infinity p = 0.0328* p = 0.0128             p = 0.047   ;             (ng .Math. h/mL) PHYT < [Log10] STE Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/mg) [Log10] Turmipure > NOV > PHYT > TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001 p = 0.0009;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; [Log10] Turmipure > NOV > PHYT < TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 24 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 8 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between p = 0   0102* p = 0.0002                 p = 0.0007;     p = 0.0050; 0 and infinity TEP < PHYT < [Log10] Turmipure Turmipure Half-life (minutes)     p < 0.0001 p = 0.0   36;     p = 0.0015;     p = 0.0111;     p = 0.0002; [Log10] Turmipure > NOV < TEP > PHYT > STE STE Turmipure Turmipure Terminal elimination     p < 0.0001 p = 0.0   36;     p = 0   ;     p = 0.0111;     p = 0.0002; rate constant Turmipure > NOV > TEP < PHYT < [Log10] STE STE Turmipure Turmipure Tmax (minutes)     p < 0.0001         p < 0.0001; p = 0.0297;     p < 0.0001; p = 0.0005; [Log10] NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00045 TABLE 43 Analysis of total parent compounds and their relative sulfate and glucuronide metabolites (ng/mL) between products in the ITT population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h     p < 0.0001             p < 0.0001;     p = 0.0057; p = 0.0005; (ng .Math. h/mL) PHYT < NOV > PHYT < [Log10] STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001         p < 0.0001; p < 0.0001;     p < 0.0001; p < 0.0001; (ng .Math. h/mL) NOV > PHYT < NOV > PHYT < [Log10] STE STE Turmipure Turmipure DN of AUC0-infinity     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0142; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-infinity     p = 0.00                p = 0.0047;             (ng .Math. h/mL) PHYT < [Log10] STE Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/mg) [Log10] Turmipure > NOV > PHYT > TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001         p < 0.0001; p = 0.0022;     p < 0.0001; p = 0.0055; [Log10] NOV > PHYT < NOV > PHYT < STE STE Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 24 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 8 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. p = 0.0397* p = 0.0151                 p = 0.0447;         between 0 and TEP < infinity [Log10] Turmipure Half-life (minutes)     p = 0.0015 p = 0.0122;     p = 0.0032;                 [Log10] Turmipure < NOV < STE STE Terminal elimination     p = 0.0015 p = 0.0122;     p = 0.0032;                 rate constant Turmipure > NOV > [Log10] STE STE Tmax (minutes)     p < 0.0001         p < 0.0001; p = 0.0020;     p < 0.0001; p = 0.0099; [Log10] NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00046 TABLE 44 Analysis of total curcunninoids (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP NOV PHYT Turmipure Turmipure Turmipure Endpoint effect effect vs STE vs STE vs STE vs STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-24 h     p < 0.0001         p < 0.0001; p < 0.0001; p = 0.0014;     p < 0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure DN of AUC0-8 h     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng .Math. h/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure AUC0-8 h     p < 0.0001 p = 0.0012;     p < 0.0001; p < 0.0001; p < 0.0001; p = 0.0048; p < 0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure DN of AUC0-infinity p = 0.0222* p < 0.0001 p = 0.0003;     p < 0.0001;     p < 0.0001;     p = 0.00   ; (ng .Math. h/mL/mg) Turmipure > NOV > TEP < PHYT < [Log10] STE STE Turmipure Turmipure AUC0-infinity     p = 0.0007             p = 0.0032;         p = 0.0014; (ng .Math. h/mL) PHYT < PHYT < [Log10] STE Turmipure Normalized Cmax     p < 0.0001 p < 0.0001;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; (ng/mL/mg) Turmipure > NOV > PHYT > TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure Cmax (ng/mL)     p < 0.0001 p = 0.0009;     p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; p < 0.0001; [Log10] Turmipure > NOV > PHYT < TEP < NOV > PHYT < STE STE STE Turmipure Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 24 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio. between     p < 0.0001                 p < 0.0001; p < 0.0001; p < 0.0001; 0 and 8 h TEP < NOV > PHYT < [Log10] Turmipure Turmipure Turmipure Rel. bio.     p < 0.0001                 p < 0.0001; p     0.0001; p < 0.0001; between 0 and TEP < NOV > PHYT < infinity [Log10] Turmipure Turmipure Turmipure Half-life (minutes)     p < 0.0001 p = 0.0062;     p = 0.002   ;     p = 0.0335;     p = 0.0010; [Log10] Turmipure < NOV < TEP > PHYT > STE STE Turmipure Turmipure Terminal elimination     p < 0.0001 p = 0.013   ;     p = 0.0034;     p = 0.0058;     p = 0.000   ; rate constant Turmipure > NOV > TEP < PHYT < [Log10] STE STE Turmipure Turmipure Tmax (minutes)     p < 0.0001         p < 0.0001; p = 0.0282;     p < 0.0001; p = 0.0002; [Log10] NOV < PHYT > NOV < PHYT > STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00047 TABLE 45 Analysis of curcumin (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) TEP vs NOV vs PHYT vs Visit Product Turmipure PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect GOLD ™ vs STE TEP vs STE NOV vs STE STE GOLD ™ GOLD ™ GOLD ™ DN of AUC0-24 h p = 0.0002 p = 0.0009; p =    ; p = 0.0010; (ng .Math. h/mL/mg) Turmipure     STE NOV     STE TEP     [Log10] Turmipure AUC0-24 h (ng .Math. h/mL/mg) [Log10] DN of AUC0-8 h p = 0.0107* (ng .Math. h/mL/mg) [Log10] AUC0-4 h (ng .Math. h/mL/mg) [Log10] DN of AUC0-infinity p = 0.0209* (ng .Math. h/mL/mg) [Log10] AUC0-infinity (ng .Math. h/mL/mg) [Log10] Normalized Cmax p = 0.0126* (ng .Math. h/mL/mg) [Log10] Cmax (ng/mL) [Log10] Rel. bio. between 0 and 24 h [Log10] Rel. bio between p = 0.0128 p =    ; 0 and 8 h TEP     [Log10] Turmipure Rel. bio. between 0 and infinity [Log10] Half-life (minutes) [Log10] Terminal elimination rate constant [Log10] Tmax (minutes) [Log10] indicates data missing or illegible when filed

TABLE-US-00048 TABLE 46 Analysis of curcumin glucuronide (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0001; p  0.0002; p  0.0001; p  0.0002; (ng .Math. h/mL) NOV > PHYT < NOV > PHYT < [Log10] STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0120; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) NOV > PHYT > NOV > PHYT < [Log10] STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT > TEP < NOV > (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) [Log10] Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0042; Cmax Turmipure > NOV > TEP < NOV > PHYT < (ng/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0401; p  0.0001; p  0.0004; (ng/mL) NOV > PHYT < NOV > PHYT < [Log10] STE STE Turmipure Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; p  0.0001; between TEP < NOV > 0 and infinity Turmipure Turmipure [Log10] Half-life p  0.0001 p  0.0157; p  0.0002; p  0.0132; (minutes) Turmipure < NOV < PHYT > [Log10] STE STE Turmipure Terminal p  0.0001 p  0.0015; p  0.0001; p  0.0123; elimination Turmipure > NOV > PHYT < rate constant STE STE Turmipure [Log10] Tmax p  0.0001 p  0.0047; p ; (minutes) NOV < NOV < [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00049 TABLE 47 Analysis of curcumin sulfate (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p = 0.027 p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p ; p  0.0003; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p = 0.0024 (ng .Math. h/mL) [Log10] DN of p = 0.028 p  0.0001 p  0.0001; p  0.0001; p  0; p  0.0001; p  0.001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p = 0.0153 p  0.0001 (ng .Math. h/mL) [Log10] DN of p = 0.03 p  0.0001 p  0.0013; p  0.0001; p  0.0014; AUC0-infinity Turmipure > NOV > TEP < (ng .Math. h/mL/mg) STE STE Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) [Log10] Normalized p = 0.02 p  0.0001 p  ; p  0.0001; p  0.0011; p  0.0001; p  0.0253; Cmax Turmipure > NOV > TEP < NOV > PHYT < (ng/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] Cmax p = 0.0001 p  0.0001 p  0.0017; p  0.0022; (ng/mL) NOV > NOV > [Log10] STE Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0002; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p = 0.0294 p  0.0001 between 0 and infinity [Log10] Half-life (minutes) [Log10] Terminal p  0.0122 elimination rate constant [Log10] Tmax p  0.0001 p  0.0001; p  ; p  0.0001; p  0.0033; (minutes) NOV > TEP > NOV < PHYT > [Log10] STE Turmipure Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00050 TABLE 48 Analysis of curcumin and its relative sulfate and glucuronide matabolites (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p = 0.043 p   p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  ; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p = 0.03 p  0.0326 (ng .Math. h/mL) [Log10] DN of p = 0.0294 p  0.0001 p  0.0001; p  0.0001; p  0.026; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) NOV > PHYT > NOV > PHYT < [Log10] STE STE Turmipure Turmipure DN of p = 0.0025 p  0.0001 p  0.0001; p  0.0001; p  0.0338; p  0.0001; p  0.0493; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-infinity p = 0.001 (ng .Math. h/mL) [Log10] Normalized p = 0.0164 p  0.0001 p  0.018; p  0.0001; p  0.0015; p  0.0001; Cmax Turmipure > NOV > TEP < NOV > (ng/mL/mg) STE STE Turmipure Turmipure [Log10] Cmax p = 0.0 p  0.0001 p  0.0001; p  0.0001; (ng/mL) NOV > NOV > [Log10] STE Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p - 0.0047 p  0.0001 p  0.0001; between TEP < 0 and infinity Turmipure [Log10] Half-life p  0.0001 p  ; (minutes) PHYT < [Log10] Turmipure Terminal p  0.0001 p  0.0202; p  0.0027; elimination NOV > PHYT < rate constant STE Turmipure [Log10] Tmax p  0.0001 p  0.0001; p  0.0001; (minutes) NOV < NOV < [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00051 TABLE 49 Analysis of DMC glucuronide (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0001; p  0.0001; (ng .Math. h/mL) NOV > NOV > [Log10] STE Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0138; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; (ng .Math. h/mL) NOV > NOV > [Log10] STE Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0143; p  0.0001; p  0.0107; AUC0-infinity Turmipure > NOV > PHYT > TEP < NOV > (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity (ng .Math. h/mL) [Log10] Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0068; p  0.0001; p  0.0001; p  0.0312; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0001; (ng/mL) NOV > NOV > [Log10] STE Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; between TEP < NOV > 0 and 24 h Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0216; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0003; between TEP < 0 and infinity Turmipure [Log10] Half-life p  0.0050 (minutes) [Log10] Terminal p  0.0002 p  0.0006; p  0.0276; elimination NOV > NOV > rate constant STE Turmipure [Log10] Tmax p  0.0086 (minutes) [Log10] indicates data missing or illegible when filed

TABLE-US-00052 TABLE 50 Analysis of DMC sulfate (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.001; AUC0-24 h Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0068; (ng .Math. h/mL) NOV < [Log10] STE DN of p = 0.0310 p  0.0001 p  0.0001; p  0.0001; p  0.0002; p  0.0001; p  0.0231; AUC0-8 h Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-8 h (ng .Math. h/mL) [Log10] DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0021; AUC0-infinity Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity p  0.0016 p  0.0021; (ng .Math. h/mL) NOV < [Log10] STE Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0016; p  0.082; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax (ng/mL) [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0015; between TEP < PHYT < 0 and 24 h Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; between TEP < PHYT < 0 and 8 h Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; between TEP < 0 and infinity Turmipure [Log10] Half-life (minutes) [Log10] Terminal elimination rate constant [Log10] Tmax p  0.0001 p  0.0001; p  0.0023; (minutes) NOV < NOV < [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00053 TABLE 51 Analysis of DMC and its relative sulfate and glucuronide metabolites (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0035; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h (ng .Math. h/mL) [Log10] DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0078; p  0.0057; (ng .Math. h/mL) NOV > NOV > [Log10] STE Turmipure DN of p  0.0001 p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT > TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity p  0.0138 (ng .Math. h/mL) [Log10] Normalized p  0.0001 p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0072; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0001; (ng/mL) NOV > NOV > [Log10] STE Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0191; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.001; p  0.0047; between TEP < PHYT < 0 and infinity Turmipure Turmipure [Log10] Half-life p  0.0004 p  0.0016; p  0.0018; (minutes) NOV < NOV < [Log10] STE Turmipure Terminal p  0.0002 p  0.0040; p  0.0002; elimination NOV > NOV > rate constant STE Turmipure [Log10] Tmax p  0.0001 p  0.0001; p  0.0073; p  0.0150; p  0.0030; p  0.0032; (minutes) NOV < PHYT > TEP > NOV < PHYT > [Log10] STE STE Turmipure Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00054 TABLE 52 Analysis of BDMC glucuronide (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0261; AUC0-24 h NOV > NOV > (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-24 h p  0.0003 p  0.0003; p  0.0341; (ng .Math. h/mL) Turmipure < NOV > [Log10] STE Turmipure DN of p  0.0001 p  0.0001; p  0.0201; AUC0-8 h NOV > NOV > (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0367; p  0.0001; p  0.0158; p  0.0001; (ng .Math. h/mL) Turmipure < TEP < PHYT < TEP > NOV > [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  ; p  0.0009; AUC0-infinity NOV > NOV > (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-infinity p  0.0003 p  0.0089; p  0.0475; p  0.0202; p  0.0422; (ng .Math. h/mL) Turmipure < PHYT < TEP > NOV > [Log10] STE STE Turmipure Turmipure Normalized p  0.0001 p  0.0001; p  0.0009; p  0.0601; Cmax NOV > TEP < NOV > (ng/mL/mg) STE Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001 p  0.0035; p  0.0001; p  0.0001; (ng/mL) Turmipure < TEP < PHYT < NOV > [Log10] STE STE STE Turmipure Rel. bio. p  0.0001 p  0.0337; p  ; between TEP < NOV > 0 and 24 h Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; between NOV > 0 and 8 h Turmipure [Log10] Rel bio. p  0.0383 between 0 and infinity [Log10] Half-life p  0.0001 p  0.0024; p  0.0003; p  0.0137; p  0.0272; (minutes) Turmipure < PHYT < TEP > NOV > [Log10] STE STE Turmipure Turmipure Terminal p  0.0001 p  0.0050; p  0.0174; p  0.0003; p  0.0001; elimination Turmipure < TEP < PHYT < NOV > rate constant STE STE STE Turmipure [Log10] Tmax p  0.0079 p  1.0176; (minutes) NOV > [Log10] Turmipure indicates data missing or illegible when filed

TABLE-US-00055 TABLE 53 Analysis of BDMC sulfate (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0012 p  0.6931; AUC0-24 h TEP < (ng .Math. h/mL/mg) Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0001; p  0.0001; p  0.0081; p  ; p  0.0057; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > PHYT > [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0151; p  ; AUC0-8 h NOV > TEP < (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > [Log10] STE STE STE Turmipure DN of AUC0-infinity (ng .Math. h/mL/mg) [Log10] AUC0-infinity p = 0.0428 p  0.0105 p  0.0305; (ng .Math. h/mL) Turmipure < [Log10] STE Normalized p = 0.0213 p  0.0248 Cmax (ng/mL/mg) [Log10] Cmax p = 0.01 p  0.0029 p  0.0109; p  0.00952; p  0.0108; (ng/mL) Turmipure < NOV < PHYT < [Log10] STE STE STE Rel. bio. p  0.0035 between 0 and 24 h [Log10] Rel. bio. p  0.0003 p  0.033; between TEP < 0 and 8 h Turmipure [Log10] Rel bio. between 0 and infinity [Log10] Half-life p  0.0001 p  0.0001; p  0.0382; p  0.0001; (minutes) Turmipure < PHYT < TEP > [Log10] STE STE Turmipure Terminal p  0.0013 p  0.0024; elimination NOV > rate constant STE [Log10] Tmax p  0.0182 p  0.0474; p  0.0147; (minutes) TEP > PHYT > [Log10] Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00056 TABLE 54 Analysis of BDMC and its relative sulfate and glucuronide metabolites (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0053; p  0.0002; AUC0-24 h NOV > TEP < (ng .Math. h/mL/mg) STE Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0001; p  0.0001; p  0.0022; p  0.0002; p  0.013; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP > PHYT > [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0139; p  0.0001; p  0.0001; p  0.0453; AUC0-8 h Turmipure > NOV < TEP < NOV > (ng .Math. h/mL/mg) STE STE Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0065; (ng .Math. h/mL) Turmipure < NOV < PHYT < TEP < NOV > [Log10] STE STE STE Turmipure Turmipure DN of p  0.0041 p  0.0334; AUC0-infinity PHYT > (ng .Math. h/mL/mg) STE [Log10] AUC0-infinity p  0.0001 p  ; p  0.0001; p  0.0157; (ng .Math. h/mL) Turmipure < NOV < TEP > PHYT > [Log10] STE STE Turmipure Turmipure Normalized p  0.0001 p  0.0031 p  0.0212; p  0.0001; p  0.0001; p  0.0001; p  0.0005; Cmax Turmipure > TEP < NOV > TEP > NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.2081 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng/mL) Turmipure < TEP < NOV > PHYT < TEP > NOV > [Log10] STE STE STE STE Turmipure Turmipure Rel. bio. p  0.0001 p  0.042; between TEP < 0 and 24 h Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; between TEP < 0 and 8 h Turmipure [Log10] Rel bio. between 0 and infinity [Log10] Half-life p  0.0001 p  0.0157; p  0.0237; p  0.0001; p  0.0308; (minutes) Turmipure < NOV < TEP > PHYT > [Log10] STE STE Turmipure Turmipure Terminal p  0.0001 p  0.0001; p  0.0007; elimination NOV > NOV > rate constant STE Turmipure [Log10] Tmax p  0.0001 p  0.0299; p  0.0943; (minutes) PHYT > PHYT > [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00057 TABLE 55 Analysis of THC glucuronide (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0211; p  0.0001; p  0.0215; p  0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0009; p  0.0001; p  0.0016; p  0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001 p  0.0001; p  0.0387; p  0.0; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT < TEP < PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure [Log10] AUC0-infinity p  0.0001; p  0.0171; p  0.0001; (ng .Math. h/mL) PHYT < TEP < PHYT < [Log10] STE Turmipure Turmipure Normalized p  0.0001 p  0.0001 p  0.0001; p  0.0182; p  0.0001; p  0.0001; p  0.0001; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0001; p  0.0252; p  0.0001; (ng/mL) NOV > PHYT > TEP < PHYT < [Log10] STE STE Turmipure Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0029; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; p  0.0001; between TEP < PHYT < 0 and infinity Turmipure Turmipure [Log10] Half-life p  0.0001 p  0.0001; p  0.0001; (minutes) PHYT < PHYT < [Log10] STE Turmipure Terminal p  0.0054 elimination rate constant [Log10] Tmax p  0.0001 p  0.0003; p  0.0003; (minutes) PHYT < PHYT < [Log10] STE Turmipure indicates data missing or illegible when filed

TABLE-US-00058 TABLE 56 Analysis of THC sulfate (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p = 0.00 AUC0-24 h (ng .Math. h/mL/mg) [Log10] AUC0-24 h p = 0.0306 (ng .Math. h/mL) [Log10] DN of p = 0.0115 AUC0-8 h (ng .Math. h/mL/mg) [Log10] AUC0-8 h p  0.0023 p  ; p  0.0003; (ng .Math. h/mL) PHYT < PHYT < [Log10] STE Turmipure DN of p  0.0029 p  ; AUC0-infinity NOV > (ng .Math. h/mL/mg) STE [Log10] AUC0-infinity p  0.0125 (ng .Math. h/mL) [Log10] Normalized p = 0.0004 p  0.0438 Cmax (ng/mL/mg) [Log10] Cmax p  0.0005 p  0.0204; p  0.005; (ng/mL) PHYT < PHYT < [Log10] STE Turmipure Rel. bio. p  0.0023 between 0 and 24 h [Log10] Rel. bio. p  0.0001 p  0.0041; p  0.0121; between TEP < PHYT < 0 and 8 h Turmipure Turmipure [Log10] Rel bio. between 0 and infinity [Log10] Half-life p  0.0098 (minutes) [Log10] Terminal p  0.0349 p  0.0151; elimination PHYT < rate constant Turmipure [Log10] Tmax p = 0.0261 (minutes) [Log10] indicates data missing or illegible when filed

TABLE-US-00059 TABLE 57 Analysis of HHC glucuronide (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  ; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0010; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) Turmipure > NOV < PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0005; p  0.0001; AUC0-infinity Turmipure > NOV > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] AUC0-infinity p  0.0001 p  ; p  0.0001; p  0.0032; p  0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; Cmax Turmipure > NOV > TEP < NOV > PHYT < (ng/mL/mg) STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0002; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure Rel. bio. p=0.0491 p  0.0001 p  0.0001; p  0.0001; between TEP < PHYT < 0 and 24 h Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; p  0.0327; p  0.0001; between TEP < NOV > PHYT < 0 and infinity Turmipure Turmipure Turmipure [Log10] Half-life (minutes) [Log10] Terminal elimination rate constant [Log10] Tmax p  0.0001 p  0.0001; p  0.0395; p  0.0001; p  0.0001; (minutes) NOV < PHYT > NOV < PHYT < [Log10] STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00060 TABLE 58 Analysis of HHC sulfate (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0510; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-infinity p  0.0001 p  0.0322; p  0.0001; p  0.0186; p  0.0001; (ng .Math. h/mL) NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0252; p  0.0001; p  0.0001; p  0.0001; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and infinity Turmipure Turmipure Turmipure [Log10] Half-life p  0.0001 p  0.0011; p  0.0012; p  0.0075; (minutes) Turmipure < NOV < TEP < [Log10] STE STE Turmipure Terminal p  0.0012 p  0.0116; p  0.0129; p  0.0458; elimination Turmipure > NOV > TEP < rate constant STE STE Turmipure [Log10] Tmax p  0.0001 p  0.0001; p  0.0001; p  0.0009; p  0.0001; (minutes) NOV < PHYT > NOV < PHYT < [Log10] STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00061 TABLE 59 Analysis of curcumin and all its relative metabolites (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0435; p  0.0001; p  0.0001; p  0.0025; p  0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < PHYT < [Log10] STE STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0007; p  0.0001; p  0.0001; p  0.0001; p  0.0070; p  0.0001; (ng .Math. h/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure DN of p = 0.0203 p  0.0001 p  0.0002; p  0.0001; p  0.0981; p  0.0034; AUC0-infinity Turmipure > NOV > TEP < PHYT < (ng .Math. h/mL/mg) STE STE Turmipure Turmipure [Log10] AUC0-infinity p  0.0001 p  ; p  0.0481; p  0.0001; p  0.0011; p  0.0001; (ng .Math. h/mL) TEP NOV > PHYT < TEP < PHYT < [Log10] STE STE Turmipure Turmipure Normalized p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.001; p  0.0001; Cmax Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0009; p  0.0001; p  0.0001; p  0.0001; p  0.001; p  0.0001; (ng/mL) Turmipure > NOV > PHYT < TEP < NOV > PHYT < [Log10] STE STE STE Turmipure Turmipure Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p  0.0001 p  0.0001; p  0.001; p  0.0001; between TEP < NOV > PHYT < 0 and infinity Turmipure Turmipure Turmipure [Log10] Half-life p  0.0001 p  0.0036; p  0.0015; p  0.0112; p  0.0002; (minutes) Turmipure < NOV < TEP > PHYT > [Log10] STE STE Turmipure Turmipure Terminal p  0.0001 p  0.0037; p  0.0009; p  0.0014; p  0.0001; elimination Turmipure > NOV > TEP < PHYT < rate constant STE STE Turmipure Turmipure [Log10] Tmax p  0.0001 p  0.0001; p  0.0298; p  0.0001; p  0.0005; (minutes) NOV < PHYT > NOV < PHYT > [Log10] STE STE Turmipure Turmipure indicates data missing or illegible when filed

TABLE-US-00062 TABLE 60 Analysis of total parent compounds and their relative sulfate and glucuronide metabolites (ng/mL) between products in the PP population. Between-group analysis-Statistical significance of comparisons between products (adjusted p-value, Tukey adjustment) Turmipure TEP vs NOV vs PHYT vs Visit Product GOLD ™ TEP vs NOV vs PHYT vs Turmipure Turmipure Turmipure Endpoint effect effect vs STE STE STE STE GOLD ™ GOLD ™ GOLD ™ DN of p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; p  0.0001; AUC0-24 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-24 h p  0.0001 p  0.0001; p  0.0002; p  0.0003; (ng .Math. h/mL) PHYT < NOV > PHYT < [Log10] STE Turmipure Turmipure DN of p = 0.0310 p  0.0001 p  0.0001; p  0.0001; p  0.0002; p  0.0001; p  0.0001; p  0.0001; AUC0-8 h Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-8 h p  0.0001 p  0.0001; p  0.0001; p  0.0001; p  0.0001; (ng .Math. h/mL) NOV > PHYT < NOV > PHYT < [Log10] STE STE Turmipure Turmipure DN of p  0.0001 p  0.0001; p  0.0001; p  0.0003; p  0.0001; p  0.0001; p  0.0001; AUC0-infinity Turmipure > NOV > PHYT > TEP < NOV > PHYT < (ng .Math. h/mL/mg) STE STE STE Turmipure Turmipure Turmipure [Log10] AUC0-infinity p  0.0072 p  0.0042; (ng .Math. h/mL) PHYT < [Log10] STE Normalized p = 0.0228 p  0.0001 p  0.0018; p  0.0001; p  0.0014; p  0.0001; Cmax Turmipure > NOV > TEP < NOV > (ng/mL/mg) STE STE Turmipure Turmipure [Log10] Cmax p  0.0001 p  0.0001; p  0.0020; p  0.0001; p  ; (ng/mL) NOV > PHYT < NOV > PHYT < [Log10] STE STE Turmipure Turmipure Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 24 h Turmipure Turmipure Turmipure [Log10] Rel. bio. p  0.0001 p  0.0001; p  0.0001; p  0.0001; between TEP < NOV > PHYT < 0 and 8 h Turmipure Turmipure Turmipure [Log10] Rel bio. p = 0.0362 p  0.0213 between 0 and infinity [Log10] Half-life p  0.0019 p  0.0125; p  0.0040; (minutes) Turmipure < NOV < [Log10] STE STE Terminal p  0.0003 p  0.0181; p  0.0056; p  0.0135; elimination Turmipure > NOV > PHYT < rate constant STE STE Turmipure [Log10] Tmax p  0.0001 p  0.0001; p  0.0008; p  0.0001; p  0.0047; (minutes) NOV < PHYT > NOV < PHYT > [Log10] STE STE Turmipure Turmipure indicates data missing or illegible when filed

[0497] Serious Adverse Events: [0498] Subject SN01-009: Cervical pain/road accident between V0 and V1 visits (under no study product) (locomotor/rheumatologic body system, event not related to a medical history, moderate intensity, no action on the study product, event not related to the research and the study product, event not associated with corrective treatments, recovery without sequelae). [0499] Treatment emergent AEs with severe intensity: [0500] Subject SN01-007: Lumbago between V2 and V3 visits (under Turmipure GOLD™ product) (locomotor/rheumatologic body system, event not related to a medical history, severe intensity, no action on the study product, event not related to the research and the study product, event not associated with corrective treatments, recovery without sequelae). [0501] AEs related to the study products: [0502] Subject SN01-008: Headache the day of V3 visit (under Turmipure GOLD™ product) (neurologic/psychiatric body system, event not related to a medical history, moderate intensity, no action on the study product, possible event related to the research and the study product, event associated with a corrective treatment (paracetamol), recovery without sequelae). [0503] Subject SN01-030: Headache the day of V1 visit (under Turmipure GOLD™ product) (neurologic/psychiatric body system, event not related to a medical history, mild intensity, no action on the study product, possible event related to the research and the study product, event associated with a corrective treatment (paracetamol), recovery without sequelae). [0504] Subject SN01-032: Headache the day of V2 visit (under NOV product) (neurologic/psychiatric body system, event not related to a medical history, mild intensity, no action on the study product, possible event related to the research and the study product, event not associated with corrective treatments, recovery without sequelae).

[0505] The results observed on PP population are similar to those on ITT population.

[0506] Mean±SD for Total curcuminoids (ITT population) is shown below.

TABLE-US-00063 TABLE 61 Mean ± SD for Total curcuminoids (ITT population) STE TEP NOV PHYT Turmipure GOLD ™ product product product product product Dose-normalized AUC(0-24 h) (ng .Math. h/mL/mg) 3.7 (1.75) 3.2 (1.69) 136.1 (37.40)  13.0 (9.65)  72.9 (25.49) AUC(0-24 h) (ng .Math. h/mL/mg) 5075.6 (2407.80) 4382.3 (2328.43) 8538.8 (2345.94) 2327.1 (1727.61) 6519.7 (2280.33) Dose-normalized AUC(0-8 h) (ng .Math. h/mL/mg) 1.6 (0.81) 1.2 (0.52) 81.8 (19.17) 4.1 (3.83) 38.1 (12.74) AUC(0-8 h) (ng .Math. h/mL/mg) 2204.2 (1111.03) 1725.7 (723.46)  5132.6 (1202.41) 737.6 (686.44) 3410.0 (1139.65) Dose-normalized AUC (0-infinity) 5.3 (5.80) 3.2 (1.68) 149.4 (64.93)  14.6 (15.02) 78.3 (37.82) (ng .Math. h/mL/mg) AUC(0-infinity) (ng .Math. h/mL/mg) 7273.2 (7969.82) 4370.3 (2317.23) 9369.3 (4072.30) 2618.8 (2689.04) 7001.1 (3382.63) Normalized Cmax (ng/mL/mg) 0.3 (0.14) 0.3 (0.10) 28.1 (7.23)  1.2 (0.80) 7.6 (3.06) Cmax (ng/mL) 444.7 (192.96) 372.8 (141.70) 1762.9 (453.72)  209.1 (144.10) 678.0 (273.35) Relative bioavailability between 0 and 24 h 1.0 (0.00) 1.1 (0.79) 49.7 (37.78) 4.2 (4.15) 24.2 (15.48) Relative bioavailability between 0 and 8 h 1.0 (0.00) 1.1 (1.07) 72.2 (58.68) 2.9 (2.50) 30.6 (20.00) Relative bioavailability between 0 and infinity 1.0 (0.00) 1.0 (0.75) 49.3 (37.74) 3.7 (4.13) 22.9 (15.28) Half-life (minutes)  788.4 (1233.91) 505.2 (265.35) 337.3 (278.83) 640.1 (399.12) 318.3 (154.44) Terminal elimination rate constant 0.1 (0.05) 0.1 (0.04) 0.2 (0.05) 0.1 (0.05) 0.2 (0.06) Time to peak (minutes) 256.5 (181.69) 330.3 (341.21) 61.0 (18.45) 375.0 (249.39) 189.5 (147.52)

CONCLUSION

[0507] The results demonstrate that there are few differences between the bioavailability of the compounds found in TEP and STE (only 5 differences).

[0508] The composition within the scope of the present invention (Turmipure GOLD™) was found to provide better bioavailability of compounds than STE, TEP and PHYT, and was able to provide a similar bioavailability to NOV despite being administered at a lower dose (300 mg compared to 1000 mg) and using only natural ingredients with no synthetic carriers (such as polysorbate 80).

[0509] More specifically, Novasol was used at 1000 mg whereas the composition of the present invention (Turmipure GOLD™) was at 300 mg. As Turmipure yields an effect of 6520 (AUC in ng.Math.h/mL) at 300 mg, if it were used at the same dosage as Novasol (1000 mg), it would yield an effect of 21733 (AUC in ng.Math.h/mL), which is much higher than the effect of Novasol at the same dosage (8539).