Claims:WE CLAIM:

1. A composition of curcumin comprising a molecular complex of curcumin and a nutraceutical, wherein the curcumin has a water solubility of at least 0.1 µg/ml.

2. The composition as claimed in claim 1, wherein the nutraceutical is selected from a group comprising amino acids, peptides, proteins, carotenoids, minerals, polyphenols, flavonoids, vitamins, sterols, stanols, polysaccharides and dietary fiber.

3. The composition as claimed in claim 1, wherein curcumin and the nutraceutical are in a stoichiometric ratio in a range of 1: 5 to 5:1.

4. The composition as claimed in claim 1, wherein curcumin and the nutraceutical are in a stoichiometric ratio of 1: 1.

5. A process for increasing the bioavailability of curcumin, the process comprising preparing a molecular complex of curcumin and a nutraceutical by
mixing curcumin and the nutraceutical in a stoichiometric ratio in a range of 1: 5 to 5: 1 followed by grinding to obtain a dry mixture;
adding a solvent to the dry mixture followed by grinding to obtain a wet mixture; and
air drying the wet mixture to obtain the molecular complex of curcumin and the nutraceutical wherein curcumin has a water solubility of at least 0.1 µg/ml.

6. The process as claimed in claim 5, further comprising heating the wet mixture at about 70 ºC prior to drying.

7. The process as claimed in claim 5, wherein the wet mixture is heated under a nitrogen atmosphere.

8. The process as claimed in claim 5, wherein the solvent is added to the dry mixture in a range of 0.5 ml to 10 ml with respect to each gram of combined weight of curcumin and the nutraceutical.

9. The process as claimed in claim 5, wherein the solvent is selected from water, acetonitrile, ethanol, methanol, ethyl acetate, acetone and mixtures thereof.

10. The process as claimed in claim 5, wherein the solvent is added to the dry mixture in a range of 0.1 ml to 5 ml per 1 gram of combined weight of curcumin and the nutraceutical.

Dated this 28th day of December, 2016

Aparna Kareer
Of Obhan & Associates
Agent for the Applicant
Patent Agent No. 1359 , Description:FIELD OF INVENTION
The present disclosure relates to a molecular complex of curcumin and a nutraceutical and a process for production thereof.

BACKGROUND
Curcumin is a bright yellow pigment, which is a primary active constituent of turmeric, a commonly used spice, derived from the rhizomatous herbaceous plant Curcuma Longa. The herbaceous perennial plant belongs to the ginger family. Curcumin is widely used as a therapeutic agent in diseases such as inflammatory bowel disease, peptic ulcer, pancreatitis, arthritis, cancer, skin wounds, etc. However, curcumin has poor absorption and rapid metabolism rate, which limits its bioavailability. Hence, there is need to enhance the bioavailability of curcumin.

SUMMARY
A nutraceutical formulation of curcumin and a nutraceutical is disclosed. A nutraceutical formulation comprises a molecular complex of curcumin and a nutraceutical. The molecular complex has improved water solubility and improved bioavailability.
A process for increasing the bioavailability of curcumin is also disclosed. The process comprises preparing a molecular complex of curcumin and a nutraceutical by mixing curcumin and the nutraceutical in a stoichiometric ratio in a range of 1: 5 to 5: 1 followed by grinding to obtain a dry mixture; adding a solvent to the dry mixture followed by grinding to obtain a wet mixture; and air drying the wet mixture to obtain the molecular complex of curcumin and the nutraceutical wherein curcumin has a water solubility of at least 0.1 µg/ml.

BRIEF DESCRIPTION OF DRAWINGS
The accompanying figures illustrate the preferred embodiments and together with the following detailed description explain the principles of disclosed process and product.
Figure 1 illustrates a comparison powder X-ray diffraction (PXRD) profile of molecular complex of curcumin and green tea extract in a stoichiometric ratio of 1:1.
Figure 2 illustrates the comparison infrared (IR) absorption profile of various samples of molecular complex of curcumin and green tea extract in a stoichiometric ratio of 1:1.
Figure 3 illustrates the comparison Differential Scanning Calorimetry (DSC) profile of various samples of molecular complex of curcumin and green tea extract in a stoichiometric ratio of 1:1.
Figure 4 illustrates a comparison powder X-ray diffraction (PXRD) profile of molecular complex of curcumin and ferulic acid in a stoichiometric ratio of 1:2.
Figure 5 illustrates the comparison infrared (IR) absorption profile of various samples of molecular complex of curcumin and ferulic acid in a stoichiometric ratio of 1:2.
Figure 6 illustrates the comparison Differential Scanning Calorimetry (DSC) profile of various samples of molecular complex of curcumin and ferulic acid in a stoichiometric ratio of 1:2.
Figure 7 illustrates the standard known concentration curve of curcumin solubility using Ultra Violet-visible spectroscopy.

DETAILED DESCRIPTION
For the purpose of promoting an understanding of the principles of the invention, reference will now be made to embodiments and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the disclosed product and process, and such further applications of the principles of the invention therein being contemplated as would normally occur to one skilled in the art to which the invention relates.
It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to be restrictive.
Reference throughout this specification to “one embodiment” “an embodiment” or similar language means that a particular feature, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase “in one embodiment”, “in an embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
Definitions
Molecular Complex: herein refers to a substance in a solid form or solid formulation, which comprises at least two pure substances which interact with each other through hydrogen bonding or any other non-covalent interactions to form molecular complex including co-crystals, solvates, hydrates or eutectic combinations or solid solutions, in which at least one of the pure substance is present in the solid form.
Nutraceuticals also known as phytochemicals, are natural, bioactive chemical compounds that provide numerous physiological benefits, including, inter alia, disease/illness prevention/treatment and health promotion. Nutraceuticals are used to achieve both long-term and short-term health objectives.
Bioavailability: The U.S Food and Drug Administration (FDA) define bioavailability as “the rate and extent to which the active drug ingredient or therapeutic moiety is absorbed from a drug product and becomes available at the site of drug action”. Because in practice it is rare that active ingredient concentrations can be determined at the site of action.
The therapeutic effectiveness of an active ingredient depends upon the ability of the dosage form to deliver the active ingredient to its site of action at a rate and amount sufficient to elicit the desired pharmacological response. This attribute of the dosage form is referred to as physiologic availability or simply bioavailability.
For most active ingredients, the pharmacologic response can be related directly to the plasma levels. Thus, the term bioavailability is defined as the rate and extent (amount) of absorption of unchanged active ingredient from its dosage form, it can also be defined as the rate and the extent to which the ingredients or active moiety is absorbed from the product and becomes available at the site of action. Blood level studies are the most common type of human or animal bioavailability studies, and are based on the assumption that there is a direct relationship between the concentration of active ingredient in blood and or plasma and the concentration of active ingredient at the site of action. By monitoring the concentration in the blood, it is thus possible to obtain an indirect measure of active ingredient response. Following the administration of a single dose of an active ingredient, blood samples are drawn at specific time intervals and analyzed for active ingredient content. A profile is constructed showing the concentration of active ingredient in blood at the specific times the samples were taken. The key parameters to note are:
1. AUC, the area under the plasma concentration-time curve, The AUC is proportional to the total amount of active ingredient reaching the systemic circulation, and thus characterizes the extent of absorption.
2. Cmax, the maximum active ingredient concentration. The maximum concentration of active ingredient in the plasma is a function of both the rate and extent of absorption. Cmax will increase with an increase in the dose, as well as with an increase in the absorption rate.
3. Tmax, the time at which Cmax occurs. The Tmax reflects the rate of active ingredient absorption, and decreases as the absorption rate increases.
Bioavailability (the rate and extent of active ingredient absorption) is generally assessed by the determination of these three parameters. Since the AUC is representative of, and proportional to, the total amount of active ingredient absorbed into the circulation, it is used to quantitate the extent of active ingredient absorption. Thus, the faster the absorption of an active ingredient the higher the maximum concentration will be and the less time it will take to reach the maximum concentration.
Improved solubility: as employed herein refer to a form of the curcumin that has increased solubility or dissolution rate relative to the least soluble form of the curcumin known.
The disclosure relates to a molecular complex of curcumin and a nutraceutical. In accordance with an aspect, the water solubility of curcumin in the curcumin and nutraceutical molecular complex is at least 0.1 microgram per milliliter (µg/ml) of water. The water solubility was checked in substantially pure water. The maximum water solubility of the curcumin has been reported 11 Nano gram per milliliter by Ucisik M H et al (Journal of Nanobitotechnology 2013, 11:37).
Chemically, curcumin is a bis-R, -unsaturated-diketone (commonly called diferuloymethane), which exhibits keto-enol tautomerism having a predominant keto form in acidic and neutral solutions and stable enol form in alkaline medium. Commercial, curcumin contains approximately 77% diferuloymethane, 17% demethoxycurcumin, and 6% bisdemethoxycurcumin.
Curcumin refers to all polymorphs, solvates, and hydrates of the all curcuminoids, having the formula I, II and III, alone or in combination in any ratio:



Nutraceuticals refer to phytochemicals, that are natural, bioactive chemical compounds that provide numerous physiological benefits, including, inter alia, disease/illness prevention/treatment and health promotion. Nutraceuticals are used to achieve both long-term and short-term health objectives.
In accordance with an embodiment, nutraceuticals are selected from a group comprising amino acids, peptides, proteins, carotenoids, minerals, polyphenols, flavonoids, vitamins, sterols, stanols, polysaccharides and dietary fiber.
By way of an example amino acids may include but are not limited to arginine, aspartic acid, glutamic acid, glutamine, valine and proline. By way of an example peptides may include but are not limited to dipeptides, tripeptides, polypeptides, phenolic phytochemicals, ferulic acid, chlorogenic acid and caffeic acid. By way of an example carotenoids may include but are not limited to lutein, beta-carotene, lycopene and astaxanthin. By way of an example minerals may include but are not limited to calcium and magnesium. By way of an example polyphenols may include but are not limited to ellagic acid, tannic acid, theaflavins, epicatechin, epicatechin-3-gallate, epigallocatechin, catechin, gallocatechin, green tea polyphenols extract and pomegranate polyphenols extract.
By way of a specific example, the nutraceutical is green tea extract.
By way of another specific example, the nutraceutical is ferulic acid.
In accordance with an aspect, the stoichiometric ratio of curcumin and the nutraceutical in the molecular complex is any stoichiometric ratios between 1: 5 to 5: 1, preferably 1:1.
In accordance with an aspect, molecular complex of curcumin and the nutraceutical has a differential scanning calorimetry melting peak in the range of 146ºC to 175ºC. Differential scanning calorimetry analysis determines the temperature and heat flow associated with material transitions as a function of time and temperature. The change in the differential scanning calorimetry melting peak determines the formulation of molecular complex. By way of a specific example, molecular complex of curcumin and green tea extract has a differential scanning calorimetry melting peak in the range of 161 ºC to 163 ºC. By way of another specific example, molecular complex of curcumin and ferulic acid has a differential scanning calorimetry melting peak in the range of 146 ºC to 149 ºC.
In accordance with an aspect, molecular complex of curcumin and the nutraceutical has a change in thermal behavior in case of formation of an amorphous material. In accordance with an aspect, the thermal behavior is change in glass transition temperature.
In accordance with an aspect, the nutraceutical formulation comprising curcumin and nutraceutical has improved bioavailability. In accordance with an aspect the molecular complex of curcumin and the nutraceutical has 250% of bioavailability compared to curcumin as 100% bioavailability.
In accordance with an aspect, the invention provides a pharmaceutical, foodstuff, nutritional supplement, and nutritional composition comprising molecular complex of curcumin and nutraceutical as described herewith.
The disclosure also provides a process for increasing the bioavailability of curcumin. The bioavailability of curcumin is increased by preparing a molecular complex of curcumin and the nutraceutical.
The process comprises of preparing a mixture of curcumin and the nutraceutical in different molar stoichiometric ratios, grinding the mixture, heating the ground mixture, cooling the heated mixture and grinding it to obtain molecular complex of curcumin and nutraceutical.
In accordance with an embodiment, nutraceuticals are selected from a group comprising amino acids, peptides, proteins, carotenoids, minerals, polyphenols, flavonoids, vitamins, sterols, stanols, polysaccharides and dietary fiber.
By way of an example amino acids may include but are not limited to arginine, aspartic acid, glutamic acid, glutamine, valine and proline. By way of an example peptides may include but are not limited to dipeptides, tripeptides, polypeptides, phenolic phytochemicals, ferulic acid, chlorogenic acid and caffeic acid. By way of an example carotenoids may include but are not limited to lutein, beta-carotene, lycopene and astaxanthin. By way of an example minerals may include but are not limited to calcium and magnesium. By way of an example polyphenols may include but are not limited to ellagic acid, tannic acid, theaflavins, epicatechin, epicatechin-3-gallate, epigallocatechin, catechin, gallocatechin, green tea polyphenols extract and pomegranate polyphenols extract.
By way of a specific example, the nutraceutical is green tea extract.
By way of another specific example, the nutraceutical is ferulic acid.
In accordance with an embodiment, the ground mixture is heated at 60 to 20 degree Celsius. The ground mixture may be heated using any known heating device including but not limited to rotary evaporator, temperature control water bath with or without vacuum. The heating may be carried out under an inert atmosphere.
The wet grinding and heating may be carried out under nitrogen atmosphere.
The grinding of the dry mixture may be carried out for 1 to 20 minutes. The grinding of the wet mixture may be carried out for 1 to 20 minutes.
The grinding may be carried out in any suitable apparatus for grinding solids. Such apparatus includes but is not limited to mortar mills, vibrator mills or ball mills.
In accordance with an embodiment, the solvent is any suitable solvent including but not limited to water, acetonitrile, ethanol, methanol, ethyl acetate, acetone or their mixture. The amount of solvent added is in a range of 0.1 ml to 5 ml per 1 gram of combined weight of curcumin and the nutraceutical.
By way of a specific example the process comprises of admixing curcumin and nutraceutical in a 1: 1 stoichiometric ratio to form a dry mixture, grinding the dry mixture for a predetermined period of time, adding few drops of solvent to make it wet and grinding the wet mixture for a predetermined period of time under inert condition, heating this ground mixture at around 75 degree Celsius temperature in a water bath for a period of 10 to 15 minutes under inert condition and cooling the heated ground mixture to room temperature to obtain the molecular complex of curcumin and the nutraceutical.
In accordance with an embodiment, the composition comprises the molecular complex of curcumin and the nutraceutical of the present disclosure as an active ingredient may be provided as a pharmaceutical composition for preventing or treating inflammation, skin wounds, pain relief, indigestion or another common ailment. The composition of the present disclosure may comprise molecular complex of curcumin and the nutraceutical in an amount of 0.001 – 99.999% (w/w) as well as an excipient as balance.
In accordance with an embodiment, the nutraceutical formulation comprising the molecular complex of curcumin and the nutraceutical of the present disclosure may be formulated into powder, granule, tablet, sugar-coated tablet, capsule, liquid, gel, syrup, suspension, wafer, or the like together with an adequate excipient for oral administration according to a method known in the art.
In accordance with an embodiment, the nutraceutical formulation comprising the molecular complex of curcumin and nutraceutical of the present disclosure is a food additive.

SPECIFIC EMBODIMENTS ARE DESCRIBED BELOW
A composition of curcumin comprising a molecular complex of curcumin and a nutraceutical, wherein the curcumin has a water solubility of at least 0.1 µg/ml.
Such a composition, wherein the nutraceutical is selected from a group comprising amino acids, peptides, proteins, carotenoids, minerals, polyphenols, flavonoids, vitamins, sterols, stanols, polysaccharides and dietary fiber.
Such a composition, wherein curcumin and the nutraceutical are in a stoichiometric ratio in a range of 1: 5 to 5:1.
Such a composition as claimed in claim 1, wherein curcumin and the nutraceutical are in a stoichiometric ratio of 1: 1.
A process for increasing the bioavailability of curcumin, the process comprising preparing a molecular complex of curcumin and a nutraceutical by mixing curcumin and the nutraceutical in a stoichiometric ratio in a range of 1: 5 to 5: 1 followed by grinding to obtain a dry mixture; adding a solvent to the dry mixture followed by grinding to obtain a wet mixture; and air drying the wet mixture to obtain the molecular complex of curcumin and the nutraceutical wherein curcumin has a water solubility of at least 0.1 µg/ml.
Such a process, further comprising heating the wet mixture at about 70 ºC prior to drying.
Such a process, wherein the wet mixture is heated under a nitrogen atmosphere.
Such a process, wherein the solvent is added to the dry mixture in a range of 0.5 ml to 10 ml with respect to each gram of combined weight of curcumin and the nutraceutical.
Such a process, wherein the solvent is selected from water, acetonitrile, ethanol, methanol, ethyl acetate, acetone and mixtures thereof.
Such a process, wherein the solvent is added to the dry mixture in a range of 0.1 ml to 5 ml per 1 gram of combined weight of curcumin and the nutraceutical.

EXAMPLES
Example 1: Molecular Complex Preparation
Curcumin (Sigma-Aldrich Co, USA) and green tea extract (Shanghai Novanat Co. Ltd, China) were weighed in 1:1, 1:2, 3:1 and 4:1 stoichiometric ratios (where extract molar mass was calculated based on different % of catechin content) as separate samples for different experiments and further ground using mortar and pestle for 5 minutes to make a homogenous mixture followed by making a paste under inert condition (nitrogen atmosphere) with a few drops of water added to it. The mixture was heated to around 70 °C temperature in a control water bath for 5-10 min under vacuum in a rotary evaporator. The resulted material was cooled to room temperature to obtain the molecular complex.
Example 2:
Curcumin (Sigma-Aldrich Co, USA) and ferulic acid (Sigma-Aldrich Co, USA) were weighed in 1:1, 1:2 stoichiometric ratios as separate samples for different experiments and further ground using mortar and pestle for 5 minutes to make a homogenous mixture followed making a paste under inert condition (nitrogen atmosphere) with few drops of water added to it. The mixture was heated to around 70 °C temperature in a control water bath for 5-10 min under vacuum in a rotary evaporator. The resulted material was cooled to room temperature to obtain the molecular complex.
Example 3: Powder X-Ray Diffraction
Technical Details:
Powder X-ray Diffraction (PXRD) profiles were obtained using PANalytical “X”pert PRO diffractometer. PXRD profile data presented for the region where significant peaks were observed.
Analysis:
Molecular complexes of curcumin and green tea extract gave different PXRD profile than pure curcumin and green tea extract. Figure 1 illustrates a comparative PXRD profile of samples of molecular complexes of Curcumin and green tea extract of stoichiometric ratio 1:1. Figure 4 illustrates a comparative PXRD profile of samples of molecular complexes of Curcumin and Ferulic acid of stoichiometric ratio 1:2
The difference in PXRD profile between the samples of pure curcumin and green tea extract and the molecular complexes of curcumin and green tea extract indicate that a molecular complex of curcumin and green tea extract are formed using the disclosed process. Different PXRD profile Curcumin-Ferulic acid molecular complex than pure curcumin and ferulic acid indicate formation of molecular complex.
Example 4: Infrared Spectroscopy
Technical Details:
Fourier transformed infrared spectra (FT-IR) were collected on a Bruker Vertex 70 model.
Analysis:
FT-IR spectra of all the molecular complexes obtained from examples 1 and example 2 were compared with individual compounds and it was found that there are significant changes in IR spectral band of functional group regions to confirm the formation of novel molecular complexes. Figure 2 and Figure 5 exhibits the major IR peaks of the sample of molecular complexes of curcumin and green tea extract of stoichiometric ratio 1:1 and curcumin and green tea extract of stoichiometric ratio 1:2.
Example 5: Differential Scanning Calorimetry
Technical Details:
Differential Scanning Calorimetry (DSC) thermograms of all the samples including samples from Example 1 were recorded on a Mettler DSC1 instrument.
Analysis:
Molecular complexes of curcumin-green tea extract and curcumin-ferulic acid gave completely different DSC profile. Figure 3 and Figure 4 illustrates the comparative DSC profile of samples of molecular complexes of curcumin-green tea extract of stoichiometric ratio 1:1 and curcumin-ferulic acid of stoichiometric ratio 1:1.
Example 6: Solubility Study of Molecular Complex
Water solubility of curcumin in molecular complex of curcumin and green tea extract was checked in water. 150 mg of molecular complexes of curcumin and green tea extract with 1:1 stoichiometric ratio was weighed and transferred in flasks containing 50 mL water. Flask was kept on shaker at 100 round per minute (RPM) of speed. 2 ml aliquot was withdrawn separately from the flasks time to time and at the time interval of 24 hours for equilibrium stage followed by the aliquots were filtered through 0.45 µ PTFE filter. Filtered samples were analyzed using Carry5000 UV/Vis instruments at 420 nm for the determination of curcumin concentration, which provide solubility of curcumin formulations. Water solubility of curcumin in molecular complexes of curcumin and green tea extract was determined from the known concentration standard curve of curcumin in methanol as illustrated with Figure 7. The curcumin solubility of different samples in water has been enumerated in table 1.

Table 1: UV Absorbance of pure Curcumin in known concentration
Concentration (µg/ml) Absorbance
1.6 0.205
2.4 0.286
3.2 0.392
4 0.482
4.8 0.594
5.6 0.663
8 0.978

Table 2: Water Solubility study of molecular complex
Sr. No. Molecular Complex (Curcumin and Green Tea Extract Ratio) Curcumin water Solubility (24 hours)
µg/ml
1 1:1 1.84

Table 1 and 2 indicate that the solubility of curcumin increases when it is present as a molecular complex.
Example 7: Bioavailability Study of Molecular Complex in Rat Model
Bioavailability (pharmacokinetics) of curcumin test was conducted as per standard guidelines. Pure curcumin as control sample and molecular complexes of curcumin and ferulic acid (1:2) as exemplary product was taken as test sample for the bioavailability study. The study was conducted by orally feeding to male Sprague Dawley rats. Rats were taken in two groups the first group was for control group and the second group was for test sample group. In each group three male Sprague Dawley rats were taken for study. For control group dose of 300 mg/kg of curcumin was administrated to each rat and similarly an equivalent dose of 300 mg/kg of curcumin was administrated to each rat in test sample group. Blood samples were collected at pre-dose and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h post-dose from each animal within ± 2 min of schedule time. At each time point, approximately 0.25 ml of blood was withdrawn and transferred to a labeled microfuge tube containing 200 mM K2EDTA solution (20 µL per mL of blood). Following sampling, equal volume of heparinized saline was replaced into the catheter. Blood was processed to collect plasma and stored below -60 °C until their bioanalysis. Each plasma sample was analyzed for curcumin using a fit-for purpose liquid chromatographic tandem mass spectrometric detection (LC-MS/MS) method with a lower limit of quantification of 0.5 ng/mL for curcumin. The pharmacokinetic parameters for free curcumin were calculated using the non-compartmental analysis tool of the validated Phoenix WinNonlin software (Version 6.3). Mean pharmacokinetic parameters of curcumin-ferulic acid formulation and standard curcumin are given in below Table 3.
Table 3: Mean pharmacokinetic parameters of Curcumin
Treatment/
Group Sample Tmax
(h) Cmax
(ng/mL) AUClast
(ng.h/mL) Relative bioavailability
Control group Curcumin 6.0
(4.0 – 6.0) 9.09
±
1.88 35.8
±
5.61 NA
Test sample group Curcumin-Ferulic acid 6.0
(6.0 – 6.0) 26.1
±
6.21 222
±
88.9 620%

The results of Table 3 indicate that the molecular complex of curcumin-ferulic acid has increased bioavailability.

INDUSTRIAL APPLICABILITY
The composition as disclosed provides better solubility of curcumin in water and better nutraceutical stability. As curcumin has better solubility and is stable, this allows enhanced bioavailability of curcumin. The enhanced bioavailability leads to better absorption in body for better bio-efficacy of the active ingredient. Further, the molecular complex of curcumin and the nutraceutical has synergistic effect.
The composition disclosed comprising the molecular complex of curcumin and the nutraceutical as an active ingredient may be provided as a pharmaceutical composition for preventing or treating inflammation, skin wounds, pain relief, indigestion or another common ailment. The composition of the present disclosure comprises molecular complex of curcumin and the nutraceutical in an amount of 0.001 – 99.999% (w/w) as well as an excipient as balance.