TECHNICAL FIELD

The present disclosure relates to a composition comprising extracts of ginger and vanilla with Curcumin for increasing bioavailability of Curcumin. The composition is found to be helpful in inhibiting the growth of tumor. The disclosure also provides method for obtaining the said composition.

BACKGROUND

Research in the latter half of the 20th century has identified that curcumin has a wide range of potential therapeutic or preventive effects. So far, these effects have not been confirmed in humans. However, as of 2008, numerous clinical trials in humans were underway, studying the effect of Curcumin on numerous diseases including multiple myeloma, pancreatic cancer, myelodysplastic syndromes, colon cancer, psoriasis and Alzheimer's disease.

In vitro and animal studies have suggested the curcumin may have antitumor, antioxidant, antiarthritic, anti-amyloid, anti-ischemic, and anti-inflammatory properties. Anti¬inflammatory properties may be due to inhibition of eicosanoid biosynthesis. In addition it may be effective in treating malaria, prevention of cervical cancer and may interfere with the replication of the HIV virus. In HIV, it appears to act by interfering with P300/CREB-binding protein (CBP). It is also hepatoprotective.

A 2008 study at Michigan State University showed that low concentrations of Curcumin interfere with Herpes simplex virus-1 (HSV-1) replication. The same study showed that Curcumin inhibited the recruitment of RNA polymerase II to viral DNA, thus inhibiting the transcription of the viral DNA. This effect was shown to be independent of effect on histone acetyltransferase activities of p300/CBP. A previous (1999) study performed at University of Cincinnati indicated that Curcumin is significantly associated with protection from infection by HSV-2 in animal models of intravaginal infections.

Curcumin acts as a free radical scavenger and antioxidant, inhibiting lipid peroxidation and oxidative DNA damage. Curcuminoids induce glutathione S-transferase and are potent inhibitors of cytochrome P450. A 2004 UCLA-Veterans

Affairs study involving genetically altered mice suggests that curcumin might inhibit the accumulation of destructive beta-amyloid in the brains of Alzheimer's disease patients and also break up existing plaques associated with the disease.

There is also circumstantial evidence that Curcumin improves mental functions; a survey of 1010 Asian people who ate yellow curry and were between the ages of 60 and 93 showed that those who ate the sauce "once every six months" or more had higher MMSE results than those who did not. From a scientific standpoint, though, this does not show whether the curry caused it, or people who had healthy habits also tended to eat the curry, or some completely different relationship.

Numerous studies have demonstrated that Curcumin, amongst only a few other things such as high impact exercise, learning, bright light, and antidepressant usage, has a positive effect on neurogenesis in the hippocampus and concentrations of brain-derived neurotrophic factor (BDNF), reductions in both of which are associated with stress, depression, and anxiety. Curcumin has also been demonstrated to be a selective monoamine oxidase inhibitor (MAOI) of type MAO-A.

In 2009, an Iranian group demonstrated the combination effect of curcumin with 24 antibiotics against Staphylococcus aureus. It is showed that in the presence of sub inhibitory concentration of curcumin the antibacterial activities of cefixime, cefotaxime, vancomycin and tetracycline have been increased against test strain. Its potential anticancer effects stem from its ability to induce apoptosis in cancer cells without cytotoxic effects on healthy cells. Curcumin can interfere with the activity of the transcription factor NF-KB, which has been linked to a number of inflammatory diseases such as cancer.

A 2009 study suggests that curcumin may inhibit mTOR complex I via a novel mechanism. Another 2009 study on curcumin effects on cancer states that curcumim "modulates growth of tumor cells through regulation of multiple cell signaling pathways including cell proliferation pathway (cyclin Dl, c-myc), cell survival pathway (Bcl-2, Bcl-xL, cFLIP, XIAP, c-IAPl), caspase activation pathway (caspase-8, 3, 9), tumor suppressor pathway (p53, p21) death receptor pathway (DR4, DR5), mitochondrial pathways, and protein kinase pathway (JNK, Akt, and AMPK)". When 0.2% curcumin is added to diet given to rats or mice previously given a carcinogen, it significantly reduces colon carcinogenesis.

The pharmacological safety and efficacy of Curcumin makes it a potential compound for treatment and prevention of a wide variety of human diseases. In spite of its efficacy and safety, Curcumin has not yet been approved as a therapeutic agent, and the relative bioavailability of Curcumin has been highlighted as a major problem for this. Major reasons contributing to the low plasma and tissue levels of Curcumin appear to be due to poor absorption, rapid metabolism, and rapid systemic elimination. Low serum level of Curcumin is one of the major factors affecting its bioavailability. A recent study showed that after oral administration of 400 mg of Curcumin to rats only traces of unchanged drug were found in the liver and kidney. At 30 min, 90% of Curcumin was found in the stomach and small intestine, but only 1% was present at 24 h. Once absorbed Curcumin get metabolized rapidly in liver and gets excreted out of the biological system. This seriously limits Curcumin to reach targets distant from the gut and exert its beneficial action.

To improve the bioavailability of Curcumin, numerous approaches have been undertaken. The reports so far show that the absorbtion, biodistribution, metabolism and elimination of Curcumin are the major problems that affect the bioavailability of Curcumin. Several works has been done to overcome these problems. Use of adjuvants (bioavailability enhancers), are one important means in this route. Other methods include nanoparticle technology, encapsulation of Curcumin in liposome, complexation of Curcumin to form micelles and phospholipids, and the introduction of bioconjugates.

Adjuvants are compounds that are concomitantly administered with the drug/nutraceutical so as to enhance the bioavailability of the latter. One of the main reasons of poor bioavailability of Curcumin is the glucuronidation in the liver. The adjuvants like piperine are shown to inhibit this metabolic pathway of glucuronidation thus increasing the bioavailability of Curcumin.

Several adjuvants are reported to enhance the bioavailability of curcumin.
Concomitant administration of Piperin along with curcumin (Bioperin) has shown to increase the bioavailability of curcumin. Piperin inhibits hepatic and intestinal glucuronidation, which is the major pathway of curcumin metabolism, thereby reducing glucuronidation of curcumin subsequently reducing the elimination from the body. The human body uses glucuronidation to make a large variety of substances more water-soluble, and, in this way, allow for their subsequent elimination from the body upon urination.

Gingerol also have been reported to enhance the bioavailability of curcumin. In a work by Khajuria et al (2003), it has been shown that gingerol enhances bioavailability of several nutraceutical drugs such as Vitamin A, E, C, Folic acid, b-carotene, Silymarin, Isoleucine, Zinc and Potassium. A comparison of the activity of gingerol to piperin and a combination of (50:50) Gingerol: piperin is also reported.

For the given dose (50mg/kg) Gingerol is shown to enhance the bioavailability of curcumin by 43% where as piperin shows the same enhancement by 33%. A combination of gingerol and piperin shows curcumin bioenhancement of 70%. The screening and comparison has been made for several immunosuppresents, herbal formulations, cortisosteroids, anti-histamines and anti-ulcer drugs which shows gingerol has a higher bioenhancing activity when compared to piperin. Other reported bioavailability enhancers of Curcumin are eugenol, EGCG, quarecetin and genisetein.

The present disclosure provides a composition comprising ginger extract and vanilla extract along with Curcumin to enhance the bioavailability of Curcumin.

STATEMENT OF DISCLOSURE

Accordingly the present disclosure provides a synergistic composition, comprising Curcumin, Vanilla extract and Ginger extract, optionally along with pharmaceutically acceptable excipients; a process for preparation of synergistic composition comprising Curcumin, Vanilla extract and Ginger extract comprising acts of, mixing the Curcumin, the Vanilla extract and the Ginger extract to obtain a mixture, and optionally adding excipients to obtain the synergistic composition and method of enhancing bioavailability of Curcumin, said method comprising act of administering biologically suitable amounts of synergistic composition of present disclosure to a subject in need thereof.

BRIEF DESCRIPTION OF ACCOMPANYING FIGURES

In order that the disclosure may be readily understood and put into practical effect, reference will now be made to exemplary embodiments as illustrated with reference to the accompanying figures. The figure together with a detailed description below, are incorporated in and form part of the specification, and serve to further illustrate the embodiments and explain various principles and advantages, in accordance with the present disclosure where,

Figure 1 shows oral pharmacokinetics studies in male Sprague dawley rats (lg/Kg body weight).

Figure 2 shows in vivo anti-cancer activity of Curcumin composition 1 in HCT-116 human xenograft model.

DETAILED DESCRIPTION OF DISCLOSURE

The present disclosure is in relation to a synergistic composition, comprising Curcumin, Vanilla extract and Ginger extract, optionally along with pharmaceutically acceptable excipient.

In an embodiment of the present disclosure, the Curcumin is present in a range of about 60% w/w to about 85% w/w, the Ginger extract is present in a range of about 5% w/w to about 25% w/w, and the Vanilla extract is present in a range of about 0.5% w/w to about 5 % w/w.

The composition as above, wherein the Curcumin is present in a range of about 84% w/w to about 86% w/w, the Ginger extract is present in a range of about 12% w/w to about 13% w/w and the Vanilla extract is present in a range of about 2% w/w to about 3% w/w.

In another embodiment of the present disclosure, the Curcumin is comprising curcuminoids in a range of about 92% w/w to about 94% w/w, the Ginger extract comprising Gingerol in a range of about 10% w/w to about 12% w/w and the Vanilla extract comprising Vanillin in a range of about 0. 5% w/w to about 2.5% w/w.

In yet another embodiment of the present disclosure, the Curcumin, the Vanillin and the Gingerol are obtained from a source selected from a group comprising plants and chemical synthesis.

In still another embodiment of the present disclosure, the excipients are selected from a group comprising granulating agents, binding agents, lubricating agents, disintegrating agents, sweetening agents, glidants, anti-adherents, anti-static agents, surfactants, anti¬oxidants, coating agents, coloring agents, flavouring agents, plasticizers, preservatives, suspending agents, emulsifying agents, spheronization agents and any combination thereof.

In still another embodiment of the present disclosure, the composition is formulated into forms selected from a group comprising tablet, troches, lozenges, aqueous or oily suspensions, capsule, emulsion, creams, spray, drops, dispersible powders or granules, syrups, elixirs, phytoceuticals and nutraceuticals.

The present disclosure is also in relation to a process for preparation of synergistic composition comprising Curcumin, Vanilla extract and Ginger extract comprising acts of,

a. mixing the Curcumin, the Vanilla extract and the Ginger extract to obtain a mixture; and

b. optionally adding excipients to obtain the synergistic composition.

In still another embodiment of the present disclosure, the Curcumin, the Vanilla extract and the Ginger extract are in a form selected from a group comprising solid and liquid.

In still another embodiment of the present disclosure, the mixing is carried out using blender selected from a group comprising ball mill mixer, sand mill mixer, multi mill mixer, pulverizer and granulator.

The present disclosure is also in relation to a method of enhancing bioavailability of Curcumin, said method comprising act of contacting cell with suitable amount of synergistic composition of claim 1 in a subject in need thereof.

In another embodiment of the present disclosure, the subject is an animal, including human being.

In an embodiment, the present disclosure provides use of a combination of ginger extract rich in gingerol (up to 35%) and vanilla extract rich in vanillin (up to 5%) which enhance the bioavailability of Curcumin. It is found that a composition (composition 1) containing ginger extract rich in gingerol (about 10%) and vanilla extract (about 0.75% rich in vanillin) with curcumin (about 94% curcuminoids), when administered orally to Sprague dawley rats, the concentration of curcumin in blood plasma seemed to have significant increase when compared to the control. The enhancement in bioavailability is found to be about 7.5 folds when compared to the control. It is also noted that there is an enhancement in the bioavailability of the Curcumin when the composition is used as compared to the bioavailability in respect to Curcumin when used with ginger extract or Vanilla extract per se.

In another embodiment of the present disclosure, apart from the enhancement of bioavailability of Curcumin with the help of the extract, it can also be noted that the composition is pleasant smelling due to the presence of Vanillin.

In another embodiment of the present disclosure, the synergistic composition of the present disclosure is also noted for enhancing the anti-cancer activity by inhibiting the tumor growth.

The details of method of preparation of the composition are given below. However it should not be construed that the scope disclosure is limited to the examples.

Example 1:

Materials used to prepare compositions:

A. Curcumin: Curcumin is the turmeric extract with total curcuminoids of 80-94%. Curcumin powder is extracted from turmeric rhizomes (Curcuma Longa) using ethyl acetate as a solvent. The extracted oleoresin is refluxed with a non-polar solvent (hexane) for removing the oil content present in the curcumin extract. Further purification is done by crystallizing from a suitable mixture of ethyl acetate and hexane. The final crystallization is done using a alcohol selected from a group comprising isoproplyl, isobutyl, neopentyl alcohol to get the total curcuminoids of 94 % [Curcumin, demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC)].

Henceforth the Curcumin means the turmeric extract with total curcuminoids of 92-94%.

B.Ginger extract: The ginger extract is rich in gingerol. Ginger raw material (RM) of Burma variety is used for SCF-C02 (Supercritical fluid extraction using C02) extraction after sun drying.

The RM is loaded into the extractor and the extraction is carried for 3 hours at 250 bar pressure and 45 °C. The top note is collected the separator and Gingerol rich ginger extract is collected in separately. The ginger extract is centrifuged to remove any moisture content which is in resin form having a gingerol content of 35%. The extract is mixed with maltodextrin and spray dried to get the powdered product with Gingerol content of about10 to 12%.

Henceforth ginger extract means ginger extract obtained as above mixed with maltodextrin in the powder form with Gingerol content of about 10-12%.
C.Vanilla extract: This is the plant extract obtained from A grade Vanilla beans (RM) from Karnataka.

The RM having a moisture content of 30% is extracted in aqueous alcohol and the extract obtained is spray dried after mixing with maltodextrin to a powder product with 0.75% vanillin content.

Henceforth vanilla extract means vanilla extract mixed with maltodextrin in the powder form with vanillin content of 0.75%.

In an embodiment of the present disclosure, commercially available Curcumin, Ginger extract and vanilla extract with appropriate activity also can be used.

Example 2: Preparation of compositions

D. Composition 1

42.5 g of curcumin is mixed with 6.25 g of ginger extract and 1.25g of vanilla extract in a mortar and mixed in a lab homogenizer to get 50 g of composition 1.

This can also prepared in large scale in plant level. Here 85kg of curcumin extract is blended with ginger extract 12.5 kg and 2.5 kg vanilla extract in a homogenizer to get 100 kg of the composition 1.

E. Composition 2

42.5g of curcumin is mixed with 7.5 g of ginger extract in a mortar and mixed in a lab homogenizer to get 50 g of composition 2.

F. Composition 3

44.5g of curcumin is mixed with 4.25 g of ginger extract and 1.25 g of vanilla extract in a mortar and mixed in a lab homogenizer to get 50g of composition 3.

G. Composition 4

46.5g of curcumin is mixed with 2.25g of ginger extract and 1.25g of vanilla extract in a mortar and mixed in a lab homogenizer to get 50g of composition 4.

H. Composition 5

43.25 g of curcumin is mixed with 6.25g of ginger extract and 0.5g of vanilla extract in a mortar and mixed in a lab homogenizer to get 50g of composition 5.

I. Composition 6

49 g of curcumin is mixed with 1 of vanilla extract in a mortar and mixed in a lab homogenizer to get 50 g of composition 6.

J. Composition 7

49.5g of curcumin is mixed with 0.5 g of high pure capsaicin (obtained from Merck) in a mortar and mixed in a lab homogenizer to get 50 g of composition 7.

H. Formulation

Composition 1 is filled as such into capsules (500 mg each) for oral consumption.

In another embodiment of the present disclosure, the said compositions are also obtained by mixing appropriate quantities of synthetic curcuminoids, gingerol and vanillin.

Example 3

Sprague dawley rats aged 7 to 8 weeks and weighing around 200 to 250 g were used. Animals were fasted overnight with free access to water. Animals were administered test substance by oral route with a Curcuminoids equivalent dose of 1 g /kg body weight (in a suitable formulation and dose volume). Blood samples (150-200 l) were collected at various time points during the next 24 hours (0.08, 0.25, 0.5, 1, 2, 4, 8 and 24 hours).

The blood samples were centrifuged at 3000 g for 5 min at 4 °C and the corresponding plasma samples were harvested in clean pre-labeled tubes. Analysis by ultra fast liquid chromatography (UFLC) was carried out on same day, or samples were stored at -80 °C till analysis.

Bio analytical Procedure:

Curcumin in plasma samples quantified using UFLC (SCHIMADZU) with suitable extraction and recovery methods.

Data Analysis and Report

The data is analyzed by WinNonlin (Pharsight) to calculate PK parameters: AUCo-t, AUC0-oo, Cmax, Tmax, t1/2 and MRT last

The compositions used for the pharmacokinetic studies and the active ingredients involved in compositions are summarized in table 1 and 2 respectively.

Table 1

Table 2 - The actual content of active ingredients in compositions by HPLC:

The table 3 provides the details of the pharmacokinetic studies carried out with the above mentioned compositions. From the studies it is evident that presence of ginger extract, vanilla extract and capsaicin enhances the plasma exposure of Curcumin.

As compared to Curcumin control, the composition of Curcumin (85%), Ginger extract (12.5%), Vanilla extract (2.5%) (Composition 1) resulted in 7.5 fold increase in area under the curve (AU last) and 2.3 fold increase in Cmax as detailed in figure 1.

The Curcumin, Vanilla extract (composition 6) and Curcumin, Capsaicin (composition 7) also showed increased bioavailability as compared to Curcumin control.

Table 3: Curcumin compositions: Oral pharmacokinetics studies in male Sprague dawley rats (lg /kg body weight equivalent dose)

Example 4: ANTI-CANCER ACTIVITY OF CURCUMIN COMPOSITION 1
Human colorectal (HCT-116) xenograft model in SCID mice

Five weeks old female N0D.CB17-prkdcscid/NcrCrl mice purchased from Advanced Centre for Treatment, Research and Education in Cancer (ACTREC) Mumbai were used for in vivo experiments. The in vivo study was approved by the Institutional Animal Ethics committee (IAEC) Animals were injected with lxlO6 HCT -116 cells subcutaneously in the flank region. Animals were monitored daily during the period between inoculation and palpable tumor growth. Tumor size was measured with a digital Vernier caliper and tumor-bearing mice were randomized into control and treatment groups (n=8), when the tumor volume was attained -100 mm . The tumor bearing mice were orally administered the curcumin composition 1, curcumin control and the Reference compound (Vorinostat) at the doses mentioned once daily for 28 days. Tumor volume and was body weight was measured twice weekly. The following formula was used to calculate the tumor volume: Tumor volume = (length x width2)/2. Percent reduction in tumor growth was calculated with respect to the Vehicle treated group.

Table 4: In vivo anti-cancer activity of Curcumin composition 1 in HCT-116 human xenograft model, Tumor volume (mm3), Mean, ± SE

It can be noted that the Curcumin Composition 1 evaluated for in vivo anti-cancer activity in a HCT-116 xenograft model of colorectal cancer in SCID mice showed promising results for anti-cancer activity. The formulation at 500 mg/kg dose resulted in a 36% inhibition in tumor growth compared to 27% with curcumin control after 28 days treatment. No significant change was observed in the body weight after 28 days treatment. The figure 2 provides the details of the analysis explained above.

While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.

WE CLAIM

1. A synergistic composition comprising Curcumin, Vanilla extract and Ginger extract, optionally along with pharmaceutically acceptable excipients.

2. The composition as claimed in claim 1, wherein the Curcumin is present in a range of about 60% w/w to about 85% w/w, the Ginger extract is present in a range of about 5% w/w to about 25% w/w and the Vanilla extract is present in a range of about 0.5% w/w to about 5 % w/w.

3. The composition as claimed in claim 1, wherein the Curcumin is present in a range of about 84% w/w to about 86% w/w, the Ginger extract is present in a range of about 12% w/w to about 13% w/w and the Vanilla extract is present in a range of about 2% w/w to about 3% w/w.

4. The composition as claimed in claim 1, wherein the Curcumin is comprising curcuminoids in a range of about 92% w/w to about 94% w/w, the Ginger extract comprising Gingerol in a range of about 10% w/w to about 12% w/w and the Vanilla extract comprising Vanillin in a range of about 0. 5% w/w to about 2.5% w/w.

5. The composition as claimed in claim 4, wherein the Curcumin, the Vanillin and the Gingerol are obtained from a source selected from a group comprising plants and chemical synthesis.

6. The composition as claimed in claim 1, wherein the excipients are selected from a group comprising granulating agents, binding agents, lubricating agents, disintegrating agents, sweetening agents, glidants, anti-adherents, anti-static agents, surfactants, anti¬oxidants, coating agents, coloring agents, flavouring agents, plasticizers, preservatives, suspending agents, emulsifying agents, spheronization agents and any combination thereof.

7. The composition as claimed in claim 1, wherein the composition is formulated into forms selected from a group comprising tablet, troches, lozenges, aqueous or oily suspensions, capsule, emulsion, creams, spray, drops, dispersible powders or granules, syrups, elixirs, phytoceuticals and nutraceuticals.

8. A process for preparation of synergistic composition comprising Curcumin, Vanilla extract and Ginger extract comprising acts of,

a. mixing the Curcumin, the Vanilla extract and the Ginger extract to obtain a mixture; and

b. optionally adding excipients to obtain the synergistic composition.

9. The process as claimed in claim 8, wherein the Curcumin, the Vanilla extract and the Ginger extract are in a form selected from a group comprising solid and liquid.

10. The process as claimed in claim 8, wherein the mixing is carried out using blender selected from a group comprising ball mill mixer, sand mill mixer, multi mill mixer, pulverizer and granulator.

11. A method of enhancing bioavailability of Curcumin, said method comprising act of contacting cell with suitable amount of synergistic composition of claim 1 in a subject in need thereof.