Field of Invention:
The present invention discloses maxicuma, a water soluble composition of curcuminoids for improving the bioavailability with the addition of pharmaceutical excipients particularly modified polysaccharides, sorbitan monoleate or its derivative, preferably polysorbate. The invention relates to a formulation of 5 curcuminoids prepared from turmeric rhizome with green extraction method. The curcuminoids crystals obtained are formulated with modified polysaccharides, IT19 and HICAP 100 in the current disclosure along with sorbitan monoleate or its derivative, preferably polysorbate 80, a pharmaceutical emulsifier and dispersant.
The invention discloses a novel formulation to enhance the water solubility and 10 thereby bioavailability of curcuminoids, which will lead to more efficacious therapeutic effect of the formulation disclosed.
Background of the Invention:
Curcuminoids are polyphenols consisting of curcumin, demethoxy curcumin and bis-demethoxy curcumin. Various studies has been conducted and proved the health 15 supplementing potential of curcuminoids- as anti-oxidants, anti-inflammatory, anti-coagulant, anti-spasmodic and anti-cancerogenic. Caughey et al J. Virol 2003, 77(19) stated curcuminoids as most effective neutral molecule for many diseases.
The research work carried out to improve the bioavailability and thereby its health supplementing activities as conducted by different researches is mentioned below. 20
8568815B2: The patent describes a water soluble and stable complex formed by alkyl ether derivative of gamma cyclodextrin in a ratio of 1:1 to 1:6. The water dispersible composition prepared is useful for therapy in cancer, leukaemia and myocardial infection.
US 2009123904: Describes the addition of a positively charged Cyclodextrin to 25 organic preservation solution preferably combined with anti-oxidant and pesticide residue. 2
2014/00/0903: Describes curcumin composition with enhanced bioavailability derived from turmeric rhizome.
US 7883728B2: Describes a composition for enhanced bioavailabity of curcumin 30 and purified curcuminoids useful for therapy.
WO2015025263: Describes curcumin composition, curcumin powder and ion for improving bioavailability which consist of curcumin extract and water extract.
9192644B2: Discloses curcumin formulation having enhanced bioavailabity and comprises curcuminoids, antioxidant and glucurondation inhibitor. A method of 35 treating Alzheimer’s and other related diseases by administering the curcumin formulated.
8329233B2: Discloses curcuminoids formulation with curcumin essential oil. The purified curcuminoids crystals are added volatile oil of turmeric to improve bioavailability. 40
2012/0058208A1: Discloses a composition of curcuminoids with vanilla and ginger to improve the bioavailability of the composition.
EP255787B1: The patent related to supplementation of curcuminoids with the essential oil of turmeric. Where better curcumin delivery from supplement to the blood and tissues is better able to address conditions as rheumatoid arthritis, 45 osteoarthritis and depression.
2015/0072012A1: Describes bioavailable nano formulation with non-ionic surfactant and PEG.
The formulations of curcuminoids described in prior art and literature shows potential of curcuminoids to combat various diseases. However, the poor solubility 50 of curcuminoids in water is a major limiting factor for usefulness of curcuminoids in medical applications as studied by Zhongfa 2012, Li, H. 2013, Aggarwal N.B. et al 2013, Pawar Y.B. et al 2017 and Mukherjee A. 2009. A further study carried out by Wahlstrom and Blennow reported negligible amount of absorption of curcumin
3
in blood plasma of Sprague Dawley rats upon oral administration of 1g/kg 55 curcumin. A further study Shova et al 1998, revealed that oral administration of 500mg/kg curcumin resulted in only 1% bioavailability in rat blood plasma while at 2g/kgs oral administration, 15ng/ml of curcumin was reported in blood plasma in 50min by Yang K.Y. et al, 2007 and Cheng M.T. et al 2017.
Harsha et al 2012, disclosed a formulation of curcumin containing drug to polymer 60 ratio of 1:1 to 1:5 to improve the dispersibility. Rahman et al 2009, studied the role of surfactant in the dissolution of curcumin. All these studies have been carried out to improve the dispersibility. However, the poor bioavailability of curcuminoids lead to very less absorption by the body and high rate of metabolism which leads to elimination from the body. The outcome of the studies conducted on 65 curcuminoids by above researchers showed absorption less than 1.0%.
The curcuminoids formulation currently present in the market are based on the claim of possessing high bioavailability and manufactured by adding plant derived extracts, phospholipids and plant oils. The nanoformulation has also been claimed by use of aqueous phase in combination with emulsifier. 70
To improve the bioavailability, the present disclosure provide a unique formulation of curcuminoids 40-50% as compared to the 5-20% curcuminoids of prior art. The highly bioavailable composition contain modified polysaccharides in hydrophilic aqueous phase and particle size reduction to nano level with a dyno mill.
Other References: 75
Zhongfa L, Chiu M, Wang J Chen W, Enhancement of Curcumin oral absorption and pharmacokinetics of curcuminoids and curcumin metabolites in Mice, Cancer Chemotherapy. Pharmacology. 2012, 69, 679-689.
Li, H.,Zhang N; Hao, Y; Wang Y, Jia S, Zhang H, Formulation of curcumin delivery with functionalized single walled carbon nanotube, Drug Delivery, 2013, 80 Oct 25. 4
Aggarwal N.B. ;Jai S, Nagpal D, Aggarwal NK, Mendiratta PK, Sharma KK, Liposomal formulation of curcumin attenuates seizures in different animal models, Fundam Clini Pharmacology, 2013,27,169-172.
Pawar Y.B. Purohit H., Valicherla GR, Munjal D, Novel lipid based oral 85 formulation of curcumin, Int. J. Pharm, 2012,436, 617-623.
Mukherjee A, Viswanath JK, Formulation .characterization and evaluation of curcumin loaded PLGA nanospheres, for Cancer Therapy, Anticancer Research 2009, 29, 3867-3875.
Wahlstrom B, Blennow G, A study of fate of curcumin in rat, Acta Pharmacol 90 Toxicol 1978, 43, 86.
Shova G. Joy D; Joseph T, Majjed M., Rajendra R and Srinivas PS, Influence of Piperine on pharmacokinetic of curcumin in animal models, Planta med, 1998, 64, 353-356.
Yang K Y, Lin L C; Tseng T Y , Wang S C,, Oral bioavailabity of curcumin in rat 95 and the herbal analysis from Curcuma longa, J. Chromatography, 2007,853, 183-189.
Cheng MT, Tsai TR, Lee CY, Wei YS, Elevating bioavailability of curcumin via encapsulation, J, Agri Food Chem, 2013, 61 9666-9671.
Turmeric as Immunity booster, Online Jan 21, 2017. 100
Harsha Kharkwal, Kumud Bala, DD Joshi, Deepshikha Pande Katare, bioavailability enhancement of curcuminoids using natural polymer, Der Pharmacia Lettre, 2012, 4 (6), 1698-1711.
Rahman S M H, Telny T.C, T.K Ravi and Kuppuswamy S., Role of surfactant and pH in dissolution of curcumin, In J. Pharma Sci, 2009, 71 (2) 139-142. 105
Object of the Invention: 5
The object of the present invention is to provide a water soluble composition of curcumin or curcuminoids with improved bioavailability. The present invention discloses maxicuma, a water soluble composition of curcuminoids for improving the bioavailability, with the addition of pharmaceutical excipients particularly 110 modified polysaccharides and sorbitan monoleate or its derivative, preferably polysorbate.
Brief Summary of the Invention:
Curcuminoids extracted from turmeric rhizome contain curcumin, demethoxy curcumin and bis-demethoxy curcumin. All three constituents which are main 115 ingredients have been used in many health applications. In the present disclosure, all three constituents has been described as curcuminoids and throughout the current disclosure will be mentioned as curcuminoids. At present, the work on turmeric extract and curcuminoids conducted by many researchers’ show that the absorption, metabolism and elimination of curcuminoids is a major problem which affect the 120 bioavailability thereby under utilizing its therapeutic potential. To overcome the low bioavailability of curcumin and enhancement of bioavailability, many approaches has been used. In particular, the use of excipients which enhance the bioavailability is one important area. In addition to this encapsulation, emulsification and decreasing the particle size to nano level may be helpful to 125 improve bioavailability.
It is well established that pharmaceutical excipient improve the bioavailability of various drugs/nutraceuticals. The same hypothesis was applied in the current disclosure for the formulation of curcuminoids to improve its bioavailability and thereby its health supplementing potential. This will explore the full potential of 130 curcuminoids crystals for its numerous health benefits. Therefore, to take advantage of health benefits of curcuminoids, means of enhancing its bioavailability needs to be innovated. The current investigation make an effort in this field. The disclosure provides a composition of curcuminoids and polysaccharides, wherein the excipients are present in quantity capable of improving the bioavailability to 135 6
significant level. The polysaccharides studies in the current disclosure are maltodextrin IT 19 and HICAP 100. These two excipients either in combination or alone led to at least 3.5 fold increase in the bioavailability of the curcumin from the composition without the polysaccharides and decreasing particle size to nano scale. Addition of other bioavailability enhancing excipients such as plant phospholipids 140 along with the polysaccharides and decreasing particle size to nano level lead to the bioavailability enhancement of about 10-25 fold.
The polysaccharides, plant phospholipids and sorbitan monoleate may have synergistic effect on the bioavailability improvement. The curcuminoids crystals 95% used for preparing bioavailable formulation were extracted by green extraction 145 method with ethanol which may further extract chemicals/ ingredients leading to bioavailability enhancement.
The disclosure further provides a method of preparing the bioavailable curcuminoids composition by-
 Suspending the curcuminoids 95% crystals extracted by green solvent 150 ethanol in water and homogenizing the same by high speed stirrer.
 Addition of plant phospholipids and sorbitan monoleate derivative to the curcuminoids dispersion in water.
 Separately polysaccharides are dispersed in water and mixed with emulsified phase of curcuminoids. 155
 Thereafter, the milling of the homogenous suspension prepared with a Dyno mill to a particle size of 275-500nm is carried out. The particle size reduction may further be carried out to 250-300nm, and up to the 100nm range. Mixing polysaccharides and sorbitan monoleate, phospholipid emulsion and mixing the two composition. 160
 Milling of this suspension in high speed Dyno mill is to prepare particle size in the range of 275-500nm, preferably around 250nm and more preferably around 100nm. The current disclosure reports first such application of Dyno mill for reducing the particle size of the composition. 7
 In the current disclosure, addition of polysaccharides and plant phospholipid 165 shows bioavailability enhancing properties. Use of modified polysaccharides, IT 19 and HICAP100, and Polysorbate lead to enhancement of the bioavailable properties. The excipients IT 19 and HICAP100 thereby provide a novel formulation. In other embodiment of present disclosure, a process of preparing curcuminoids composition by 170 addition of modified polysaccharides, pharmaceutical excipient and antioxidant lead to an improvement in the bioavailabity.
It further states, the bioavailability of curcuminoids formulation can be increased by dispersing the curcuminoids 95% crystals in water and adding modified polysaccharides in it. In yet another embodiment, reduction in the particle size of 175 the curcuminoids composition in Dyno mill by formulating the crystals with the excipients described above particularly modified polysaccharides and plant derived phospholipids with or without antioxidants lead to the enhanced bioavailability significantly.
For improvement of water solubility and bioavailability of curcuminoids 180 crystals, apart from addition of modified polysaccharides and Polysorbate 80, the novel way of milling in the Dyno mill for reducing the particle size to a level of 275 to 500nm in the dispersion medium led to the significant enhancement in bioavailability. The particle size of unformulated 95% curcuminoids crystals could be reduced to 300-500nm and preferably 250nm by Dyno mill. Therefore, the 185 present disclosure aimed at exploring the effect of reducing the particle size to nano level to enhance the bioavailability of 40% curcuminoids formulated composition.
The current disclosure further states that the formulation of bioavailable curcuminoids composition is carried out with 95% curcuminoids crystals isolated from turmeric rhizome using green ethanol extraction. The ethanol extraction may 190 lead to the curcuminoids crystals having synergistic effect in improving the bioavailability of the formulation along with the effect of excipients addition and reduction in particle size. The present disclosure further mentions a process of
8
preparing a bioavailable formulation of curcuminoids having strength of curcuminoids 40-50%, while the composition invented so far have curcuminoids 195 content of 10-20%. Therefore, the composition developed in current invention might reduce the dose of the drug to a significant level.
The current disclosure further rely on the green method of preparing the drug molecule from the natural herb as extraction is carried out by environment friendly green solvent ethanol. The residual solvent traces, even if present, will have 200 minimal ill impact on the health of user and may further help in solubilizing the crystals while the formulation is carried out. The application of green solvent ethanol may also extract some polar constituents from the turmeric rhizome which may have water solubility.
The current disclosure further states that the spray drying / vacuum tray drying of 205 the 95% curcuminoids crystals- formulated with modified polysaccharide and other pharmaceutical excipient applied in present invention after reducing particle size makes it more bioavailable.
The disclosure further provides a composition of curcuminoids with modified polysaccharides and plant derived phospholipids wherein the starting curcuminoids 210 is 95%. The formulated mass have curcuminoids content 40-50%, preferably 45-50% and more preferably 40% and product is named as maxicuma.
The current disclosure further states that composition described in the current work contains curcuminoids and pharmaceutical excipient in the weight ratio of 1:1 to 1:0.8. The weight ratio of modified polysaccharides ranges from 35-45% of the 215 formulated composition.
The disclosure further states that the composition after particle size reduction is dried in spray dryer, tray dryer or by any other drying method to arrive at final formulation for bioavailability enhancement. The bioavailability study of this composition was carried out in rat model. The blood samples were withdrawn from 220 0.5 hr onwards at regular interval and concentration of curcuminoids/ metabolite in blood plasma was measured. Upon administration of the formulation by gavage, the
9
plasma concentration was found to be maximum at 0.5 hr for the formulation i.e. 40% curcuminoids.
In some of the formulation prepared by current disclosure the bioavailability 225 enhancement was 3.5 fold as compared to the bioavailability of 95% curcuminoids crystals. In other formulation the bioavailabity enhancement is 10 fold and at same dose up to 26 fold as compared to the unformulated 95% curcuminoids crystals.
Detailed Description of the Invention: 230
The detailed method of preparation of bioavailable composition consist of preparing curcuminoids crystals from turmeric rhizome powder. Adding pharmaceutical excipients possessing bioavailability enhancing properties, homogenizing, milling and final drying the formulated composition. To accomplish the targeted formulation, the current disclosure consists of a product formulated 235 using curcuminoids crystals extracted from turmeric rhizome with green ethanol extraction. The ethanol extraction containing 1-20% water could help in extraction of water soluble constituents of turmeric rhizome. In some other embodiments, the ethanol extraction was carried out with ethanol having 1-20% water, preferably 1-10% and more preferably 1-5% water in ethanol. The ethanol extract is 240 desolventized to get the oleoresin which is cooled to initiate the seeding of crystals and thereby initiating the process of crystallization. This mass is crystallized with ethanol which is then filtered to get the curcuminoids crystals 95%. The crystals are dried in vacuum/tray dryer to remove the solvent and water level. The ethanol extracted composition contain curcuminoids which is a mixture of three individual 245 constituents, i.e. curcumin, demethoxy curcumin and bis-demethoxy curcumin. The composition consists of 70-86% curcumin, followed by 10-20% demethoxy curcumin and 1-6% bis- demethoxy curcumin.
The curcuminoids composition further vary from 72-84% curcumin, 10-18% demethoxy curcumin and 1-5% bis-demethoxy curcumin. 250 10
As discussed above, the curcumin from turmeric rhizome is extracted with green solvent ethanol. The extraction is carried out at a temperature of 45-60 degree celsius under stirring. The solvent is removed from the extracted mass to prepare the extract in concentrated form. The concentrated extract of curcumin is crystallized with green solvent ethanol and water. For initiating crystallization, 255 ethanol is used in a proportion 1-3:1 with respect to the concentrated mass.
As discussed above, the limitation of curcuminoids in health supplementing properties is due to its low bioavailability and very less solubility in aqueous medium and rapid metabolism in human being. To improve curcuminoids potential for health supplementing properties various researchers have carried out work for 260 improving the bioavailability.
In pursuit to make a bioavailable composition, the curcuminoids crystals 95% are mixed with modified polysaccharides and pharmaceutical excipients and dispersed in water with constant stirring to make homogenous solution/ mixture. The dispersion is charged to the Dyno mill for reducing the particle size. The operation 265 of Dyno mill is carried from 30 minutes to 120 minutes. The experimental results show that the optimum particle size is achieved on milling for 90 minutes. This could lead to improved bioavailability of the composition prepared.
A water soluble nano-formulation curcuminoids composition (maxicuma) for increasing the bioavailability of curcumin, the composition comprises curcuma 270 longa extract, wherein, 40-50% curcuminoids based on the weight of the formulated mass, 28-50% modified polysaccharides, 2-10% sorbitan ester derivative, 0.5-2.0% natural oil, 4-10% plant derived phospholipids.
In some embodiments, the weight ratio of curcuminoids crystals and polysaccharides ranges from 1:1 to 1:1.25. In some other embodiments, the weight 275 ratio of curcuminoids and polysaccharides ranges between 0.56:1 to 0.7:1. The weight ratio of curcuminoids crystals to pharmaceutical grade excipient varies from 0.56:1 to 1:0.8. Further, in some embodiments, the pharmaceutical grade excipients
11
are also added in the form of food grade antioxidant like vitamin E and ascorbyl palmitate. 280
In other embodiment, the curcuminoids crystals are added with polysaccharides and phospholipids derived from plant for formulating bioavailable curcuminoids composition. The ratio of curcuminoids crystals and plant derived phospholipids varies from 10:1 to 10:2. The phospholipids are added with the polysaccharides which are used for formulating bioavailable curcuminoids. 285
In some embodiments, the curcuminoids crystals 95% are added with sorbitan ester i.e. monoleate or its derivatives or other emulsifier used in pharmaceutical formulations. The ratio of curcuminoids crystals and sorbitan ester or its derivative used as emulsifiers varies from 10:0.5 to 10:2, more preferably 10:1. In a few other embodiments, the sorbitan ester or its derivative, preferably polysorbate 80, are 290 added either alone or in combination with plant phospholipids for formulating bioavailable curcuminoids composition.
The application of emulsifier for solubilizing lipophilic curcuminoids is well known in prior art. However, the current disclosure describes a unique proportion of curcuminoids, sorbitan monoleate and the aqueous phase which is further milled in 295 Dyno mill to prepare the bioavailable curcuminoids formulation.
In some other embodiments, the curcuminoids crystals are added to pharmaceutical excipients mentioned in the current disclosure and solubilized by heating to a temperature of 45-60 degree celsius or by sonication in a sonicator or homogenized in a high speed homogenizer. 300
In some embodiments, all ingredients of bioavailable composition are added to the water along with the curcuminoids crystals 95%, which is then emulsified at high RPM before further proceeding for particle size reduction through wet milling.
In some other embodiments, the bioavailable composition may be added with vitamin E and ascorbyl palmitate. Vitamin E and ascorbyl palmitate are added in 305 combination or alone to make the bioavailable composition. 12
To prepare the bioavailable composition of curcuminoids crystals and pharmaceutical grade excipients, particularly polysaccharides are blended in a suitable proportion by a process including:
 suspending the curcuminoids crystals 95% in 2 to 8 times of its quantity of 310 water
 adding phospholipids and sorbitan monoleate, particularly polysorbate 80 and mixing this emulsified solution with solution of modified polysaccharides in water
 pulverization of the above blended mass in a dyno mill 315
 stripping of water under heat and/or vacuum.
In some embodiments, the curcuminoids crystals 95% are blended with polysaccharides and homogenized followed by particle size reduction in dyno mill with or without adding sorbitan monoleate or its derivative, particularly polysorbate 80, to make the formulation. 320
In some other embodiments, the wet milling of curcuminoids crystals has been carried out without adding polysaccharides and sorbitan ester. In these formulation plant derived phospholipids are added at a concentration level of 2-10%.
Antioxidants may be added to the curcuminoids formulation containing curcuminoids crystals, polysaccharides and sorbitan monoleate derivative to protect 325 the curcuminoids hydrolysis and provide long term storage stability. In the current work, vitamin E or its derivative, ascorbyl palmitate, ascorbic acid either in combination or alone has been used.
The bioavailability study with animals is carried out with the powder prepared by drying the uniform blend discussed above by vacuum drying /spray drying. 330
The particle size reduction experiments was carried out in high speed Dyno mill and particle size was reduced from 1000nm to 500nm and finally to 275nm. The particle size of curcuminoids in formulated composition is more preferably reduced to 250nm. In some embodiments, the size reduction trials were carried out from 10
13
minutes to 120 minutes. The variation of particle size with the milling time in Dyno 335 mill is presented in Fig-02, while the particle size of unformulated 95% curcuminoids crystals is presented in Fig-01. The optimum particle size of 300 nm is achieved in 90 minutes of milling, more preferably the particle size is 250-275nm. The particles could further be obtained to a level of 99-100nm at 90 minutes of milling. Therefore, a milling duration of 90 minutes is optimum for obtaining the 340 desired particle size involving the Dyno mill.
The graphical presentation of particle size at different milling time as presented in Fig- 02 shows a reduction in the particle size by 100 times from the original 95% curcuminoids crystals at 90 min of milling time. Further the Fig-02 shows that a milling time of 90 minutes is desirable to reduce the particle size to 250nm. 345
The drug release profile of maxicuma (40% curcuminoids) was observed with respect to the time in Table 3. The dissolution kinetics shows more than 70% drug release in 90 minutes.
For conducting the animal studies, the animal were kept on fasting for 24 hrs before feeding the formulated composition in 0.05% CMC. All animal studies were carried 350 out with maxicuma, a 40% curcuminoids bioavailable composition, as explained in present disclosure.
In some embodiments, the curcuminoids as part of formulation is fed to the Sprague Dawley rats through gavage and the curcuminoids/metabolite is measured in blood plasma. In a similar fashion, the unformulated curcuminoids were also fed to the 355 rats.
The curcuminoids crystals and formulated bioavailable composition were dispersed in 0.05% CMC in water. Each rat was injected 100mg curcuminoids crystals and similar quantity of formulated composition in 0.05% CMC solution.
The blood sampling was conducted from 30 minutes to 24 hrs to observe the 360 curcuminoids /metabolite concentration in blood plasma. 14
The study results achieved with maxicuma could be easily understood by person working in the field and further numerous alteration could be attempted to the examples and the instruction given herein.
Examples 365
Example 1:
1 kg turmeric rhizome is powdered and extracted with ethanol containing 2-5% water in a rotatory extractor. The powdered rhizome is added to 5L ethanol containing 2-5% water at a temperature of 45-60 degree celsius for 4 hour. The curcuminoids extracted in ethanol are separated and stored. The extraction is 370 repeated another three times under same temperature and extraction duration. The solvent is removed in a rotary evaporator at a temperature 45-60 degree celsius to make a thick paste of turmeric oleoresin. The crude paste mass is 220-250g and contains 25-30% curcuminoids. The oleoresin thus obtained is mixed with 5-20% ethanol and kept under cold condition to initiate crystallization. The crystallized 375 mass is then added to another 1 to 1.2 part ethanol and further kept for 6 to 10 days to saturate the crystals in the solvent.
The crystallized mass is filtered by centrifugation and washed with mixture of ethanol: water (20:80 to 80:20) for removing the oils and waxes from the crystals.
The centrifuged crystals are dried under vacuum in tray dryers to get 95% crystals 380 of curcuminoids. The weight of dried crystals so obtained is 34 g.
Example 2:
Process of preparing 40% curcuminoids composition (maxicuma)
To improve the bioavailability, the curcuminoids crystals thus obtained are mixed with polysaccharides and the above is dispersed in sorbitan ester derivative, plant 385 phospholipid and tocopherol. 100g curcuminoids crystals 95% are weighed and 5-10 g plant derived phospholipids, 1.0-2.0g tocopherol and 8-10g Sorbitan monoleate derivative are added, then mixed properly. In another vessel/container
15
weigh 35-40g modified polysaccharides in water. Mix two solutions / mixture at high RPM and homogenize in a high speed homogenizer. This homogenized mass 390 in then charged to a Dyno mill for particle size reduction for a time period of 15 minutes to 120 minutes. The variation of particle size for milling at different time interval is presented in Fig 02. The optimum size of the particle is achieved at a milling time of 90 minute. The same is presented in Fig-02, while the particle size distribution of 95% curcuminoids unformulated crystals is presented in Fig 01. 395
Example 3:
Process of preparing 45-50% curcuminoids (maxicuma)
To further improve concentration of maxicuma crystals to 45-50% curcuminoids, weigh 55-60g curcuminoids crystals 95%, 8-10g plant derived phospholipids, 2g tocopherol and 10g Sorbitan monoleate derivative are added then mixed properly. 400 In another vessel/container weigh 30-40g modified polysaccharides in water. Mix two solutions/ mixture at high RPM and homogenize in a high speed homogenizer. Once the particles are uniformly dispersed, this mass is homogenized and then charged to a Dyno mill for particle size reduction for a time period varying from 15 minutes to 120 minutes. The dispersed mass is then dried by rotary vacuum 405 evaporation/spray drying.
Example 4:
The impact of the formulated composition against the un-formulated 95% curcuminoids crystals were studied in animal model. Formulated curcuminoids composition containing 40-50% curcuminoids were selected for animal studies. 410 Male Sprague Dawley rats with 250-300g body weight were fasted overnight and fed with the above said composition and separate group was fed unformulated 95% curcuminoids crystals. The formulated as well as un-formulated curcuminoids were suspended / dispersed in 0.05% CMC in water and fed at 300mg/kg body weight/day by oral gavage. The blood samples of animal were withdrawn po 30 415 minutes of administering the curcuminoids dose. Further the samples of blood were withdrawn after 1hrs, 2hr, 4hr, 6hr, 14hr and 24hr post curcuminoids 16
administration. The concentration of the curcuminoids / metabolites was analysed in blood plasma by GC/MS/MS analysis.
The concentration of curcuminoids / metabolite in the blood plasma is presented in 420 Fig. 4 and 5, respectively for formulated and unformulated 95% curcuminoids crystals.
Example 5:
Oral pharmacokinetics of curcuminoids and its novel formulation maxicuma (40% curcuminoids) is evaluated in following oral administration. Extent of increase in 425 plasma Cmax, Tmax and oral exposures was analysed in case of novel formulation when compared with that of curcumin at a similar dose level.
The details of bioavailabity study protocol is as below:
Rat strain / sex: Sprague Dawley / Male
Age / body weight: 7 to 8 weeks / 250-300gms 430
 Number of animals per group: n=3
 Total number of groups: 2 [2x3 = 6 rats]
 Dose of curcumin: 300mg/kg body weight/day
 Dose of novel formulation (maxicuma): 300mg/kgs body weight/day
 Test compound route of administration: Oral 435
 Formulation Vehicle: 0.05% CMC solution
 Dose volume: 10 ml/kg
 Fast/Fed: Overnight fasted
 Dosing : Single dose
 Blood samples collection: Jugular vein cannula 440
 Time points for blood collection: 9 time points (pre-dose, 0.25, 0.30, 1.0, 2.0, 4.0, 6.0, 8.0 and 24h)
 Blood samples collection: Jugular vein cannula
 Anti-coagulant: K2EDTA
 Sample analysis by discovery grade bio analytical method developed for 445 estimation of test compound in plasma using LC-MS/MS systems. 17
The pharmacokinetic study protocol is presented in Table 2. The graphical presentation of the data show 10.2 fold increase in the bioavailability of maxicuma as compared to unformulated 95% curcuminoids crystals at a dose of 450 100mg/animal. A further study of the results indicate that the bioavailability enhancement could be almost 25 times and more if same curcuminoids content for both are considered. Since for the study maxicuma as well as unformulated 95% crystals were same i.e.100mg/animal a lower increase is encountered. However, the maxicuma is only 40% curcuminoids, extrapolating this to 100mg to a similar 455 concentration to that of unformulated 95% crystals may enhance the bioavailability by 2.5 times. The animal studies further reveals that the maximum concentration of curcuminoids in the animal fed 40% bioavailable composition reaches maximum within 0.5 h of administration of this composition. The pharmacokinetic parameter like maximum curcuminoids concentration in plasma (Cmax), the time taken to 460 reach maximum concentration (Tmax) and concentration of curcuminoids in blood maxicuma and 95% curcuminoids crystals are presented in Fig-10 and 11, respectively.
A close evaluation of the study results of current disclosure shows a bioavailability enhancement of 25-26 fold if same active dose of maxicuma is fed to the animals 465 as compared to the animal fed with unformulated 95% curcuminoids. However, at a dose of 100mg/animal of present disclosure, the bioavailability enhancement is 10.2 fold for the animals given maxicuma as compared to the animals given a dose of unformulated 95% curcuminoids crystals.
Description of the Figures 470
Fig-01: Particle size of 95% curcuminoids crystals
Fig-02: Particle size of bioavailable 40% curcuminoids (maxicuma) formulation
Fig-03: Curcumin concentration absorbance curve
Fig-04: Pharmacokinetic data of bioavailable curcuminoid 40% (maxicuma)
Fig-05: Pharmacokinetic data of unformulated 95% curcuminoids crystals 475 18
Fig-06: Mean plasma concentration of curcuminoids/ metabolite
Fig-07: Curcuminoids-glucuronide concentration for bioavailable 40% curcuminoids (maxicuma)
Fig-08: Curcuminoids-glucuronide concentration for 95% curcuminoids crystals
Fig-09: Mean plasma concentration of bioavailable 40% curcuminoids (maxicuma) 480
Fig-10: Area under Curve (AUC)-Bioavailable 40% curcuminoids (Maxicuma)
Fig-11: Area Under Curve (AUC) for 95% curcuminoids crystals
19
Table 1: Curcuminoids extraction and crystallization data
Experiment No.
Rhizome Powder, g
Curcuminoids crystals, g
Curcuminoids content,%
01
500
16
95.8
02
1000
33
97.2
03
1000
34
96.0
04
5000
150
96.2
Table 2: Pharmacokinetic study protocol 485 Group Dosing Sample Type Animal No BW (g) Dose (ml) Dose/Kgs Body weight 1 100 mg/animal of 40% Curcuminoids Formulation, PO (maxicuma) Blood in K2EDTA 1 299 3.0 10.0 2 302 3.0 10.0 3 300 3.0 10.0 2 100 mg/animal of 95% Curcuminoids crystals, PO Blood in K2EDTA 4 300 3.0 10.0 5 298 3.0 10.0 6 298 3.0 10.0
Table 3: In vivo dissolution data of 40% Curcuminoids-Maxicuma
S. No.
Time (minutes)
Amount of drug release (mg/ml)
Drug release (%)
1
Initial
8.71
43.55
2
15
14.40
72.00
3
30
15.15
75.75
4
60
14.55
72.75
5
90
19.26
96.30
490 20
Table 4: Pharmacokinetic data of bioavailable 40% curcuminoids (maxicuma) composition PK Parameters
Parameter
Animal-1
Animal-2
Animal-3
Average
Standard deviation
Cmax(ng/ml)
619.3
354.0
344.4
439.2
156.0
Tmax(h)
0.50
0.25
0.50
0.42
0.14
AUClast (hr*ng/ml)
2289.9
1404.8
1842.5
1845.7
442.5 Relative Bioavailability (%) 394.7 217.9 384.8 332.5 99.4
Table 5: Pharmacokinetic data of unformulated 95% curcuminoids crystals PK Parameters
Parameter
Animal-1
Animal-2
Animal-3
Average
Standard deviation
Cmax(ng/ml)
101.3
119.4
92.1
104.3
13.9
Tmax(h)
6.00
6.00
8.00
6.7
1.2
AUClast (hr*ng/ml)
580.1
644.8
478.8
567.9
83.7 Relative Bioavailability (%) 25.3 45.9 26.0 32.4 11.7
Table 6: Comparative data of curcuminoids-glucuronide for bioavailable 40% curcuminoids (maxicuma) and unformulated 95% curcuminoids crystals 495 PK Parameters (Plasma) Curcuminoids-glucuronide (Curcuminoids 40% formulation - 100mg/rat; 300 mg/kg/day) Curcuminoids-glucuronide (Curcuminoids 95% crystals (100mg/rat; 300 mg/kg/day)
Cmax (ng/ml)
439.2 ± 156.0
104.3 ± 13.9
Tmax (h)
0.42 ± 0.14
6.7 ± 1.2 AUCLast (ng*h/ml) 1845.7 ± 442.5 567.9 ± 83.7 21
Table 7: Oral pharmacokinetic data of curcuminoids bioavailable 40% curcuminoids (maxicuma) composition Oral PK data of Curcuminoids 40% formulation –maxicuma (100 mg/rat; 300mg/kg/day) Curcuminoids-glucuronide concentrations (n=3 animals/group) Time (h) Plasma Concentrations (ng/ml) PO Mean PO1 (100mg/rat) SD Animal 1 Animal 2 Animal 3 (ng/ml)
0.25
488.4
354.0
343.6
395.4
80.7
0.50
619.3
304.5
344.4
422.7
171.4
1.00
492.8
233.6
280.9
335.7
138.0
2.00
351.9
186.7
332.9
290.5
90.4
4.00
254.4
144.6
283.2
227.4
73.2
6.00
147.4
174.0
154.7
158.7
13.7
8.00
266.2
117.8
68.6
150.9
102.8
24.00
BQL
BQL
BQL
NA
NA
Note: results are expressed in mean±SD, n=3 animals/group, serial sampling study;
Dose: 100 mg/rat; 300 mg/kg/day of curcumin Oral PK data of Unformulated 95% Curcuminoids crystals (100 mg/rat; 300 mg/kg/day) Curcuminoids-glucuronide concentrations (n=3 animals/group) Time (h) Plasma Concentrations (ng/ml) PO Mean PO2 (100mg/rat) SD Animal 4 Animal 5 Animal 6 (ng/ml)
0.25
11.2
17.6
7.3
12.0
5.2
0.50
20.0
15.2
17.8
17.7
2.4
1.00
48.9
34.7
22.6
35.4
13.2
2.00
58.6
82.1
69.9
70.2
11.8
4.00
77.7
95.6
43.0
72.1
26.8 22
Table 8: Oral pharmacokinetic data of unformulated 95% curcuminoids crystals
500
6.00
101.3
119.4
85.2
101.9
17.1
8.00
87.6
61.8
92.1
80.5
16.3
24.00
BQL
BQL
BQL
NA
NA 23

We Claim:
1. A water soluble nano-formulation curcuminoids composition (maxicuma) for increasing the bioavailability of curcumin, the composition comprises curcuma longa extract, wherein: 505
i) 40-50% curcuminoids based on the weight of the formulated mass;
ii) 28-50% modified polysaccharides;
iii) 2-10% sorbitan ester derivative;
iv) 0.5- 2.0% natural oil;
v) 4-10% plant derived phospholipids. 510
2. The composition as claimed in claim 1, wherein the curcuminoids are obtained from rhizome of curcuma longa by green extraction method.
3. A green extraction method for the extraction of curcuminoids as claimed in 515 claim 2, comprising steps of:
i) extraction of turmeric rhizome powder with ethanol at a temperature 45-60 degree celsius, preferably 45-55 degree celsius;
ii) removal of solvent to prepare concentrated mass;
iii) seeding of oleoresin by adding 5-20% ethanol with respect to 520 oleoresin weight;
iv) crystallization with hydro-alcoholic solvent ethanol containing water 5-20% of ethanol to prepare curcumin crystals;
v) filtration of crystals and drying to obtain 95% curcuminoids crystals. 525
4. A method for preparing bioavailable curcuminoids composition as claimed in claim 1 and 3, comprising the steps of:
i) preparing aqueous solution of a modified polysaccharide in range of 28-50% by weight in warm purified water about 40 530 -80 °C under stirring till clear solution is obtained;
ii) adding fat-soluble antioxidant in range of 0.5-2.0 % by weight; 24
iii) adding surfactant sorbitan monoleate derivative, preferably polysorbate, in range of 2-10 % by weight, more preferably 535 2-8%;
iv) adding plant derived phospholipid in range of 4 -10% by weight in step 1 while stirring;
v) heating the dispersed mass to 45-60 degree celsius;
vi) stirring the mixture to disperse homogenously to make a 540 clear solution, preferably, the milling carried out in a Dyno-mill for 30-120 minutes, preferably 60-90 minutes;
vii) drying of dispersed solution by spray drying or rota vapour or vacuum tray drying, preferably by spray drying;
viii) shifting and milling to prepare uniform particles. 545
5. The method as claimed in claim 4, wherein water soluble curcuminoids (maxicuma) composition is prepared by adding modified polysaccharides, preferably maltodextrin IT 19 and HICAP 100.
6. The process as claimed in claim 4, wherein sorbitan monoleate or its derivative, preferably, polysorbate 80 either alone or in combination is used for preparing bioavailable composition.
7. The method as claimed in claim 4, wherein the bioavailable curcuminoids 555 composition is prepared by addition of plant derived phospholipids to the curcuminoids crystals in addition to the sorbitan monoleate and tocopherol for enhancing the dispersibility in water.
8. The method as claimed in claim 4, wherein particle size of curcuminoids 560 crystal is reduced less than 500 nm, preferably less than 300 nm and more preferably less than 250 nm in Dyno-mill for better dispersion and solubility of particles in aqueous medium.
9. The method as claimed in claim 4, wherein curcuminoids crystal particle 565 sizing to different time interval ranging from 15 minutes to 120 minutes to 100-250 nm and preferably to 100-150 nm.
10. The method as claimed in claim 4, consisting of drying of aqueous mass in dryer, spray dryer, vacuum tray dryer and various thin film evaporation 570 techniques.
11. The bioavailable curcuminoids composition maxicuma, is further used in aqueous dispersion by dispersing in water.
12. The method as claimed in claim 4, producing bioavailable nano-formulation is further used for studying bioavailability.