FORM 2
THE PATENT ACT, 1970
(39 of 1970)
&
THE PATENT RULES, 2003
COMPLETE SPECIFICA TION
(See section 10 and rule 13)
1. Title of the Invention
MOLECULAR COMPLEXES OF EPIGALLOCATECHIN-3-GALLATE AND PROCESS FOR PRODUCTION THEREOF
2. Applicants)
Name Nationality Address
TATA CHEMICALS INDIA BOMBAY HOUSE, 24 HOMIMODY
LIMITED STREET, MUMBAI - 400001
TATA GLOBAL BEVERAGES INDIA 1 BISHOP LEFROY ROAD, KOLKATA -
LIMITED 700 020
3. Preamble to the description
PRO VISIONAL SPECIFICA TION
The following specification particularly describes the invention and the manner in which it is to
be performed.

The present disclosure relates to molecular complex of Epigallocatechin-3-gallate and process for production thereof.
BACKGROUND
Epigallocatechin-3-gallate (EGCG), the main and most important polyphenol in green tea (Camellia sinensis), has shown numerous health promoting effects. Epigallocatechin gal late belongs to the family of catechins. It contains 3 phenol rings and has very strong antixoidant properties. Unfortunately, EGCG lacks long-term stability and is very sensitive to light and heat and also possess a very astringent and bitter taste, which limits its use in food or in oral medications.
Major reactions causing the instability of EGCG are auto-oxidation and epimerization. The rates of these reactions are affected by the temperature, pH, the partial pressure of oxygen, the level of antioxidants, the concentration of EGCG. and other components of tea. This causes the reduction in bioactivity of EGCG. EGCG also get converted into other catechins like gallocatechin gallate (GCG), Epicatechin gallate (ECG), Epigallocatechin (EGC), Gallocalechin (GC) in solid form as well as in aqueous solutions.
For food, beverages and other oral applications we need stable and tastier EGCG. A stable and tasty variant of EGCG would allow higher incorporation in enhancing the polyphenols or antioxidant content without affecting the organoleptic properties.
BRIEF DESCRIPTION OF ACCOMPANYING FIGURES AND TABLES
Figure 1 illustrates the PXRD pattern of EpigalIocatechin-3-gallale, Trehalose and Epigallocatechin-3-gallate : Trehalose molecular complex.

Figure 2 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Sucralose and Epigallocatechin-3-gallate : Sucralose molecular complex.
Figure 3 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Glutamic acid and Epigallocatechin-3-gallate : Glutamic acid molecular complex.
Figure 4 illustrates the PXRD pattern of Epigallocatechin-3-gallate. Glutamine and Epigallocatechin-3-gallate : Glutamine molecular complex.
Figure 5 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Glycine and Epigallocatechin-3-gallate : Glycine Molecualr Complex.
Figure 6 illustrates the PXRD pattern of Epigallocatechin-3-gallate. Pyroglutamic acid and Epigallocatechin-3-gallate : Pyroglutamic acid molecular complex.
Figure 7 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Lipoic Acid and Epigallocatechin-3-gallate-Lipoic acid molecular complex.
Figure 8 illustrates the PXRD Pattern of Epigallocalechin-3-gallate, Carnosine and Epigallocatechin-3-gallate-Carnosine molecular complex.
Figure 9 illustrates the PXRD Pattern of Epigallocatechin-3-gallate, Maltose and Epigallocatechin-3-gallate : Maltose molecular complex.
Figure 10 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Maltitol and Epigallocatechin-3-gallate : Maltitol molecular complex.
Figure 11 illustrates the PXRD pattern of Epigallocatechin-3-gallate, Nystose and Epigallocatechin-3-gallate : Nystose molecular complex.
Figure 12 illustrates the IR spectra of Epigaliocatechin-3-gallate, Trehalose and Epigallocatechin-3-gallate : Trehalose molecular complex.

Figure 13 illustrates the IR Spectra of Epigallocatechin-3-gallate. Sucralose and Epigallocatechin-3-gallate : Sucralose molecular complex.
Figure 14. illustrates the IR Spectra of Epigallocatechin-3-gallate. Glutamic acid and Epigallocatechin-3-galiate : Glutamic acid molecular complex.
Figure 15 illustrates the IR Spectra of Epigallocatechin-3-gallate, Glutamine and Epigallocatechin-3-gallate : Glutamine molecular complex.
Figure 16 illustrates the IR Spectra of Epigallocatechin-3-gallate. Glycine and Epigallocatechin-3-gallate : Glycine molecular complex.
Figure 17 illustrates the IR Spectra of Epigallocatechin-3-gallate, Pyroglutamic acid and Epigallocatechin-3-gallate : Pyroglutamic acid molecular complex.
Figure 18 illustrates the IR Specra of Epigallocatechin-3-gallate, Lipoic Acid and Epigallocatechin-3-gallate : Lipoic acid molecular complex.
Figure 19 illustrates the IR Spectra of Epigallocatechin-3-gallate, Carnosine and Epigallocatechin-3-gallate : Carnosine molecular complex.
Figure 20 illustrates the IR Spectra of Epigallocatechin-3-gallate, Maltose and Epigallocatechin-3-gallate : Maltose molecular complex.
Figure 21 illustrates the IR Spectra of Epigallocatechin-3-gallate, Maltitol and EpigalIocatechin-3-gallate : Maltitol molecular complex.
Figure 22 illustrates the IR Spectra of Epigallocatechin-3-gallate. Nystose and Epigallocatechin-3-gallate : Nystose molecular complex.
Figure 23 illustrates the DSC profile of Epigallocatechin-3-gallate, Trehalose and Epigallocatechin-3-gallate : Trehalose molecular complex.

Figure 24 illustrates the DSC profile of Epigallocatechin-3-gallate, Sucralose and Epigallocatechin-3-gallate : Sucralose molecular complex.
Figure 25 illustrates the DSC profile of Epigallocatechin-3-gallate, Glutamic acid and Epigallocatechin-3-gallate : Glutamic acid molecular complex.
Figure 26 illustrates the DSC profile of Epigallocatechin-3-gallate. Glutamine and Epigallocatechin-3-gallate : Glutamine molecular complex.
Figure 27 illustrates the DSC profile of Epigallocatechin-3-galtate. Glycine and Epigallocatechin-3-gallate : Glycine molecualr Complex.
Figure 28 illustrates the DSC profile of Epigallocatechin-3-gallate, Pyroglutamic acid and Epigallocatechin-3-gallate : Pyroglutamic acid molecular complex.
Figure 29 illustrates the DSC profile of Epigallocatechin-3-gallate, Lipoic Acid and Epigallocalechin-3-gallate : Lipoic acid molecular complex.
Figure 30 illustrates the DSC profile of Epigallocatechin-3-gallate, Carnosine and Epigallocatechin-3-gallate : Carnosine molecular complex.
Figure 31 illustrates the DSC profile of Epigallocatechin-3-gallate. Maltose and Epigallocatechin-3-gallate : Maltose molecular complex.
Figure 32 illustrates the DSC profile of Epigallocatechin-3-gallate, Maltitol and Epigallocatechin-3-gallate : Maltitol molecular complex.
Figure 33 illustrates the DSC profile of Epigallocatechin-3-gallate, Nystose and Epigallocatechin-3-gallate : Nystose molecular complex.
Table 1 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Trehalose and Epigallocatechin-3-gallate: Trehalose molecular complex

Table 2 exhibits the IR stretching frequency of Epiga!locatechin-3-gallate, Sucraiose and Epigallocatechin-3-gallate: Sucraiose molecular complex.
Table 3 exhibits the IR stretching frequency of Epigallocatechin-3-gallale, Glutamic acid and Epigallocatechin-3-gallate: Glutamic acid molecular complex.
Table 4 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Glutamine and Epigallocatechin-3-gallate: Glutamine molecular complex.
Table 5 exhibits the IR stretching frequency of Epigallocatechin-3-gallate. Glycine and Epigallocatechin-3-gallate: Glycine.
Table 6 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Pyro glutamic acid and Epigallocatechin-3-gallate: Pyro glutamic acid molecular complex.
Table 7 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Lipoic acid and Epigallocatechin-3-gallate: Lipoic acid molecular complex.
Table 8 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Carnosine and Epigallocatechin-3-gallate: Carnosine molecular complex.
Table 9 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, Maltose and Epigallocatechin-3-gallate: Maltose molecular complex.
Table 11 exhibits the IR stretching frequency of Epigallocatechin-3-gallate, FOS (Nystose) and Epigallocatechin-3-galIate: FOS (Nyslose) molecular complex. 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 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 of the invention and are not intended to be restrictive thereof
Reference throughout this specification to "one embodiment" "an embodiment" or similar language means that a particular feature, structure, 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.
The disclosure relates to molecular complexes of Epigallocatechin-3-gallate and a molecular complex former selected from Trehalose, Sucralose, Glutamic Acid. Glutamine, Glycine. Pyroglutamic acid, Lipoic Acid, Carnosine, Maltose, Maltitol, Fructo-oligosaccharides (FOS) and mixture thereof.
"Molecular complex" herein refers to a substance which is solid form or solid formulations, which comprises in its with at least two substances which interact with each other through hydrogen bonding or any other non covalent interactions to form molecular complex including co-crystals, solvates, hydrates, eutectic combinations or solid solutions, in which at least one of the substance is present in the solid form.
Epigallocatechin-3-gallate refers to all polymorphs, solvates, and hydrates of the substance having the formula (I):


The molecular complex former compound selected from Trehalose. Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic Acid, Carnosine, Maltose, Maltitol and FOS (Nystose) refers to all polymorphs, solvates, and hydrates of the substance having the formula (II):

Fructo-oligosaccharides: n=l: kestose (GF2), n=2: nystose (GF3), n=3: fructofuranosylnystose
(GF4)
(II)
Fructo-oligosaccharides (FOS) are oligomers of fructose having degree of polymerization 3-10 with a terminal glucose moiety where the fructose are linked by beta 2-1 linkage. It includes kestose (GF2), nystose (GF3) and fructofuranosylnystose (GF4).
The powder X-ray diffraction profile of molecular complex of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine. Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) is illustrated in figures 1 to 11.

The infrared absorption profile of molecular complex of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) is illustrated in figures 12 to 22.
The thermal behavior upon heating of molecular complex of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose. Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine. Maltose, Maltitol and/or FOS (Nystose) was measured through endothermic/exothermic transition by differential scanning calorimetry is given in figures 23 to 33.
In accordance with an aspect, the stoichiometric ratio of Epigallocatechin-3-gallate: molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glulamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) in the molecular complex of Epigallocatechin-3-gallate and molecular complex former is any molar ratios between 1:48 to 4: 1, preferably 1:4 to 4:1.
In accordance with an aspect, the invention provides a pharmaceutical, foodstuff, nutritional supplement, and nutritional composition comprising molecular complex of Epigallocatechin-3-gallate and molecular complex former selected from Trehalose. Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) as described herewith.
The process comprises of preparing a mixture of Epigallocatechin-3-gallate and molecular complex former selected from Trehalose, Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) in different molar stoichiometric ratios, grinding the mixture, heating the ground mixture, cooling

the heated mixture and grinding it to obtain molecular complex of Epigallocatechin-3-gallate and molecular complex former.
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 ground mixture is heated at 60 to 80°C using but not limited to temperature control water bath with or without vacuum. The heating may be carried out under an inert atmosphere.
Any known method may be used for preparing molecular complex of Epigallocatechin-3-gallate and molecular complex former selected from Trehalose, Sucralose, Glutamic Acid. Glutamine. Glycine. Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose). By way of a specific example the process comprises of admixing Epigallocatechin-3-gallate and molecular complex former selected from Trehalose. Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose. Maltitol and/or FOS (Nystose) in a 1:1 stoichiometric ratio to form a dry mixture, grinding said 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 °C temperature water bath for a period 10 to 15 minutes under inert condition and cooling the back the heated ground mixture to room temperature to obtain the molecular complex of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine. Maltose, Maltitol and/or FOS (Nystose).
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 1-20 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 Epigallocatechin-3-gallate and molecular complex former selected from Trehalose, Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose).
EXAMPLES
Example 1: Molecular Complex preparation:
Epigallocatechin-3-gallate and molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid. Lipoic acid, Carnosine, Maltose. Maltitol and/or FOS (Nystose) were weighed in a 1:1 molar ratio for different experiments and ground using mortar and pestle for 5 minutes to make a homogenous mixture folowed making paste under inert condition (Nitrogen atmosphere) with few drops of water added to it. Heating the mixture around 70-75 °C temperature control water bath for 15-20 min under vacuum in a rotary evaporator. The resulted material was cooled to room temperature to obtain the molecular complex.
Example 2: Powder X-ray Diffraction Technical Details:

Powder X-ray Diffraction (PXRD) profiles were obtained using PANalytical "X"pertPRO diffractometer. PXRD profile data presented for the region where significant peaks were observed. Analysis:
Molecular complexes of Epigallocatechin-3-gallate and molecular complex former selected from Trehalose, Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) gave different PXRD profile than pure Epigallocatechino-gallate and molecular complex former. Figures 1 to 11 illustrate a comparative PXRD profile of samples of molecular complexes of Epigallocatechin-3-galIate with molecular complex former along with pure Epigallocatechino-gallate and pure molecular complex former.
Example 3: 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 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 novel molecular complexes. Table 1 to 11 exhibit the major IR peaks of the samples of molecular complexes of Epigallocatechin-3-gallale and molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose). Example 4: Differential Scanning Calorimetry

Technical Details:
Differential Scanning Calorimetric (DSC) thermograms of all the samples including samples from Example 1 were recorded on a Mettler DSC1 instrument. Analysis:
Molecular complexes of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose, Sucralose, Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid. Lipoic acid. Carnosine, Maltose, Maltitol and/or FOS (Nystose) gave completely different DSC profile. Figures 23 to 33 illustrate the comparative DSC profile of samples of molecular complexes of Epigallocatechin-3-gallate with molecular complex former selected from Trehalose, Sucralose. Glutamic Acid, Glutamine, Glycine, Pyroglutamic acid, Lipoic acid, Carnosine, Maltose, Maltitol and/or FOS (Nystose) along with pure Epigallocatechin-3-gallate and molecular complex former.
INDUSTRIAL APPLICABILITY
The molecular complex of Epigallocatechin-3-gallate has better stability, lesser bitterness and is tastier. A stable and tasty variant of EGCG would allow higher incorporation in enhancing the polyphenolic or antioxidant content without affecting the organoleptic properties in food, beverages and other oral applications.