DESC:1

F O R M 2
THE PATENTS ACT, 1970
(39 of 1970) &
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)

“XANTHOPHYLL COMPOSITION COMPRISING LUTEIN AND
ZEAXANTHIN WITH ENHANCED BIOAVAILABILITY”

OMNIACTIVE HEALTH TECHNOLOGIES LIMITED
An Indian Company, registered under the Indian Companies Act, 1956 having its
registered office located at T-8b, 5th Floor, Phoenix House,
A Wing, Phoenix Mills Compound, 462, Senapati Bapat Marg, Lower Parel,
Mumbai – 400013 Maharashtra, India,

The following specification particularly describes the invention and the manner in
which it is to be performed

FIELD OF THE INVENTION
The present invention mainly relates to a Xanthophyll composition comprising of
selective isomers of lutein and zeaxanthin with a pharmaceutically and/or
nutraceutically acceptable excipients and solubility enhancers, antioxidants with bioenhancing
agents enhancing the bioavailability of actives. This invention 5 specifically
relates to Xanthophyll composition comprising of selective trans-R,R lutein and
trans-R,R zeaxanthin and at least one bioavailability enhancing agent, solubility
enhancer, and an antioxidant. The invention further relates to a xanthophyll
composition comprised of more than 80% of total xanthophylls, which is comprised
10 of selective isomers such as trans-R,R lutein and trans-R,R zeaxanthin. The invention
more specifically relates to the xanthophyll composition comprised of at least 80% by
weight of trans-R,R lutein and at least 15% by weight of trans-R,R zeaxanthin, which
is prepared by process of extraction from two different plant sources using
industrially viable process. The invention further relates to the enhancement in
15 bioavailability of xanthophyll composition comprising trans-R,R lutein and trans-R,R
zeaxanthin by combination of micronization process to obtain selective range of
particle size in preferred oil as a vehicle/carrier and selective solubility enhancer,
antioxidant and bio-enhancing agent. This surprisingly improved bioavailability is
due to combined effect of micronization process to obtain selective range of particle
20 size in preferred oil as a vehicle and selective bio-enhancing agent with solubility
enhancement and antioxidant which surprisingly exerts improvement in
bioavailability. This invention further relates to xanthophyll composition comprising
trans-R,R isomers of lutein and zeaxanthin in specific ratio that can be further
formulated in different forms like beadlets, powders, oil suspensions, granules,
25 capsules, tablets, films or any other suitable oral, parenteral or topical dosage forms
using pharmaceutically or nutraceutically acceptable excipients or carriers or
mixtures thereof. The invention further relates to xanthophyll composition, which is
3
substantially free from R,S zeaxanthin and is suitable for human consumption to be
used in various eye and brain health applications.
BACKGROUND OF THE INVENTION
The natural xanthophylls are amply present in and derived from 5 various plant and
animal sources. The beneficial health effects of natural xanthophylls and their
antioxidant nature are well documented throughout scientific literature. The only
hindrance to the effective utilization of these properties by the body is the low
bioavailability and highly oxidizable nature of these nutrients. Understanding the
10 long felt need of bioavailability, helps optimize doses for wellness supplements and
drugs.
The oral bioavailability of many xanthophyll nutrients is limited due to
physicochemical and physiological processes such as low solubility in
gastrointestinal fluids; molecular structures, metabolizing enzymes causing chemical
15 transformation within the gastrointestinal tract; low epithelium cell permeability,
membrane transporters, interaction with the gut microbiota, and the like. The
bioavailability of these agents can be improved by specifically designing the
compositions that control their release, solubilization, transport, metabolism, and
absorption within the gastrointestinal tract. Various approaches have been used to
20 combat these problems of low bioavailability of the macular nutrients for their use in
the form of nutraceutical products for human or veterinary use but there is strong felt
need to improve the bioavailability. The approaches include composition comprising
of trans-R,R zeaxanthin along with trans-R,R lutein in specific weight percentage
which is prepared from extracts of two different plant sources using food grade
25 solvents and industrially viable process of extraction, treatment with alkali,
neutralization, purification and further formulating with selective particle size by
micronization and preferred carrier or vehicle with bioavailability enhancing agent
which provides a combination effect resulting in enhanced bioavailability.
4
US6075058A teaches compositions for increased bioavailability of carotenoids. The
US patent teaches a mixture of 200 micrograms of the carotenoid lutein and 125
micrograms of the carotenoid zeaxanthin, along with the 10 mg of cholesterol, 100
mg olive oil, 20 mg egg yolk phospholipid, 250 micrograms alpha-tocopherol, and
0.375 ml of 0.15 M aqueous sodium chloride 5 solution.
WO1999/047001 relates to a method of increasing the absorption of carotenoids and,
more specifically, to increasing the absorption and bioavailability of lutein and
zeaxanthin in humans and poultry by the use of lysolecithin and lecithin.
10 IN 201821036199 relates to a purified xanthophyll composition comprising (trans R
R)-lutein and (trans R,R)-zeaxanthin, which is comprised of more than 80% of total
xanthophylls and a process for the preparation thereof. Purified xanthophyll
composition is comprised of selective isomerssuch as at least 85% by weight of (trans
R,R)-lutein and at least 15% by weight of (trans R,R)-zeaxanthin. The composition is
15 prepared by process of extraction. It is industrially viable process and purified by
employing food grade polar and nonpolar solvents. This invention mainly speaks on
the process for preparation of composition and completely silent over the formulation
composition.
The inventors of the present invention have carried out rigorous experiments to
20 prepare a xanthophyll composition which is comprised of selective isomers of trans-
R,R zeaxanthin along with trans-R,R lutein, which is prepared from extraction of two
different plant sources and subjecting the xanthophyll ester rich extracts to treatment
with alcoholic alkali. The resulting reaction mixture is then subjected to extraction,
isolation and purification process with use of food grade polar and non-polar
25 solvents to selectively get at least 15% by weight of trans-R,R zeaxanthin isomer in
combination with at least 80% by weight of trans-R,R lutein in a xanthophyll
composition.
5
The present inventors have found that the enhancement in bioavailability of
xanthophyll composition comprising trans-R,R lutein and trans-R,R zeaxanthin is due
to combined effect of micronization process to obtain selective range of particle size
in preferred oil as a vehicle, selective bio-enhancing agent, solubility enhancer and
antioxidant which surprisingly exerts improvement in 5 bioavailability.
OBJECTS OF THE PRESENT INVENTION
The main objective of the present invention is to provide xanthophyll composition
10 selectively comprising of trans-R,R zeaxanthin along with trans-R,R lutein in specific
weight percentage which is prepared from extracts of two different plant sources
using food grade solvents and industrially viable process and formulating with
pharmaceutically and/or nutraceutically accepted excipients result in enhanced
bioavailability.
15
The further main objective of the present invention is an enhancement in
bioavailability of xanthophyll composition comprising trans-R,R lutein and trans-R,R
zeaxanthin is due to combined effect of micronization process to obtain selective
range of particle size in preferred oil as a vehicle with selective bio-enhancing agent,
20 solubility enhancer and antioxidant which surprisingly exerts improved
bioavailability.
The further objective of the present invention is to provide xanthophyll composition
comprising trans-R,R isomers of lutein and trans-R,R isomers of zeaxanthin in
specific ratio is further formulated in different forms like beadlets, powders, oil
25 suspensions, granules, capsules, tablets, films or any other suitable oral, parenteral or
topical dosage forms using pharmaceutically or nutraceutically acceptable excipients
and/or carriers.
6
Another objective of the present invention is to provide xanthophyll composition of
trans-R,R zeaxanthin along with trans-R,R lutein and selective bioenhancer showing
enhanced bioavailability with 90% Confidence interval for test formulation being
more statistically significant i.e. more than 80-125% criteria of bioequivalence to
show superior bioavailability compared to reference 5 formulation.
One more objective of the present invention is to provide xanthophyll composition
comprised of at least 80% of total xanthophylls, which is further comprised of at least
80% by weight of trans-R,R lutein and at least 15 % by weight of trans-R,R
10 zeaxanthin.
One more objective of the present invention is to provide xanthophyll rich
composition comprised of selective isomers such as trans-R,R zeaxanthin and trans-
R,R lutein and which is substantially free from R,S and S,S zeaxanthin .
15
Still one more objective of the present invention is to provide a composition which is
obtained from extracts of two different plant sources comprising xanthophyll esters
which are prepared by treatment with food grade solvent in specific proportions by
using specific reaction conditions with unit operations.
20
One more important objective of the present invention is to provide xanthophyll rich
composition, which is prepared from extracts of two different plant sources such as
Marigold and Paprika.
25 One more objective of the present invention is to provide a process for preparation of
xanthophyll composition which is prepared by mixing two xanthophyll ester rich
extracts in definite proportion and subjecting the reaction mixture to treatment with
alcoholic alkali, followed by neutralization isolation, and purification process to get
desired purity of xanthophyll composition.
7
One more objective of the present invention is to provide selection of particle size by
micronization and further addition of suitable oil as a vehicle with selective bioenhancer,
solubility enhancer, antioxidant and optionally flavoring agent and mixing
to obtain xanthophyll composition in the form of oil suspension.
5
One more objective of the present invention is to provide xanthophyll composition
selectively comprising of trans-R,R zeaxanthin and trans-R,R lutein along with at
least one pharmaceutically and/or nutraceutically accepted excipient to prepare
beadlets.
10
One more objective of the present invention is to provide xanthophyll composition,
which is substantially free from R,S and S,S zeaxanthin and is suitable for human
consumption to be used in various eye and brain health applications.
15 SUMMARYOF THE PRESENT INVENTION
According to an aspect of the present invention, there is provided xanthophyll
composition comprising:
i) trans-R,R lutein in the range of 65-85% by weight;
20 ii) trans-R,R zeaxanthin in the range of 10-30% by weight;
iii) one or more carrier and/or vehicle in the range of 10-80% by weight;
iv) one or more solubility enhancer in the range of 1-10% by weight
v) one or more flavoring agentin the range of 1-5% by weight;
vi) one or more bioavailability enhancing agent in the range of 1-10% by
25 weight; and
vii) one or more antioxidant agent in the range of 1-10% by weight.
8
wherein trans-R,R lutein and trans-R,R zeaxanthin extracted from marigold and
paprika oleoresin and are present in the ratio of 4:1 to 6:1, preferably 5:1, and the said
composition is substantially free from R,S and S,S zeaxanthin.
The present invention further extends to the process for preparation of formulation as
oil suspension 5 and beadlets.
BRIEF DESCRIPTION OF ACCOMPANYING FIGURES
According to figure 1-8 the graphs designated as Product A (Micro OS R+) read as
trans-R,R lutein and trans-R,R zeaxanthin composition. Reference is product B.
10
Figure 1 shows Comparative Linear Plot of Baseline Corrected Mean ± SD of Serum
trans-RR lutein Concentrations vs Time for Micro OS R+ and Reference. Figure 1
demonstrates that Baseline corrected serum trans-R,R lutein concentrations for Micro
OS R+ group were significantly higher (P<0.05) compared to Reference group
15 between 2 to 72 hrs time points post dose.
Figure 2 & 3 shows trans-R,R lutein Pharmacokinetic Parameters - Cmax, AUC0-72
& AUC0-T: Micro OS R+ Vs Reference. Figure 2 &3 demonstrate that Micro OS
R+ group showed significantly higher absorption for trans-R,R lutein compared to
20 Reference group as evident from the 2.5X folds Cmax, 2.9X folds AUC0-72 and
3.2X folds AUC0-T levels.
Figure 4 shows Comparative linear Plot of Baseline Corrected Mean ± SD of Serum
trans-R,R zeaxanthin Concentrations vs Time for Micro OS R+ and Reference.
25 Figure 4 demonstrates that Baseline corrected serum trans-R,R Zeaxanthin
concentrations for Micro OS R+ group were significantly higher (P<0.05) compared
to Reference group between 4 to 72 hrs time points post dose.
9
Figure 5 and Figure 6 shows trans-R,R zeaxanthin Pharmacokinetic Parameters -
Cmax, AUC0-72 & AUC0-T: Micro OS R+ Vs Reference. Figure 5 and 6
demonstrate that Micro OS R+ group showed significantly higher absorption for
trans-R,R zeaxanthin compared to Reference group as evident from the 1.7X folds
Cmax and 2.2X folds AUC0-5 72 & AUC0-T levels.
Figure 7 shows particle size of trans-R,R lutein and trans-R,R zeaxanthin before
preparation of oil suspension.
Figure 8 shows particle size of trans-R,R lutein and trans-R,R zeaxanthin after
preparation of oil suspension.
10
DETAILED DESCRIPTION OF PRESENT INVENTION
The present invention relates to a xanthophyll composition comprising selective
isomers such as trans-R,R lutein and trans-R,R zeaxanthin and a process for the
15 preparation thereof. The invention specifically relates to a xanthophyll composition
comprised of more than 80% of total xanthophylls, which is further comprised of at
least 80% by weight of trans-R,R lutein and at least 15% by weight of trans-R,R
zeaxanthin. The process is comprised of mixing the extracts containing xanthophylls
esters obtained from two different plant sources in definite proportion and further
20 treated to get xanthophyll composition selectively comprised of trans-R,R lutein and
trans-R,R zeaxanthin in desired percentage. The enhancement in bioavailability of
xanthophyll composition comprising trans-R,R lutein and trans-R,R zeaxanthin is due
to combined effect of micronization process to obtain selective range of particle size
in preferred oil as a vehicle and selective bio-enhancing agent, solubility enhancer
25 and antioxidant with flavoring agent which surprisingly exerts improvement in
bioavailability in 1.5 to 4 fold against the reference. This xanthophyll composition
comprising of trans-R,R isomers of lutein and trans-R,R isomers of zeaxanthin in
10
specific ratio from 4:1 to 6:1 is further formulated in different forms like beadlets,
powders, oil suspensions, granules, capsules, tablets, films or any other suitable oral,
parenteral or topical dosage forms using pharmaceutically or nutraceutically
acceptable excipients and/or carriers. The xanthophyll composition, which is
substantially free from R,S and S,S zeaxanthin and is suitable for 5 human consumption
to be used in various eye and brain health applications.
According to embodiment of the present invention the xanthophyll composition
selectively comprising of trans-R,R lutein and trans-R,R zeaxanthin in specific
10 weight percentage which is prepared from extracts of two different plant sources
using food grade solvents and industrially viable process treatment with alkali,
neutralization, isolation, purification and then formulating with pharmaceutically
and/or nutraceutically accepted excipients result in enhanced bioavailability.
15 Within the context of this invention, the term ‘xanthophyll composition’ means a
composition comprised of xanthophylls such as selective isomers of lutein and
zeaxanthin which is prepared from two different plant sources and purified by using
food grade solvents, so that the composition is selectively comprised of trans-R,R
lutein and trans-R,R zeaxanthin. More importantly the composition is substantially
20 free from R,S and S,S isomer of zeaxanthin.
According to the main embodiment of the invention, the xanthophyll composition is
selectively comprised of at least 80% by weight of total xanthophylls, of which at
least 65-85% by weight being trans-R,R lutein and at least 10-30% by weight being
25 trans-R,R zeaxanthin which is useful for nutrition and healthcare.
According to important embodiment of the invention, the plant material used for the
extraction may be selected from various sources including, but not limited to, fruits,
flowers and vegetables such as kiwi fruit, grapes, spinach, orange juice, zucchini (or
11
vegetable marrow), and different kinds of squash, paprika, other dark green leafy
vegetables, parsley, kale, egg yolk, maize and the like. As per one of the
embodiments of the invention, the plant material selected for preparation of the
composition is marigold flower (Tagetes erecta) and hybrid species of Paprika.
Particularly lutein ester rich extract is obtained from marigold flower 5 and zeaxanthin
ester rich extract is obtained from specific variety of Paprika.
As per one more embodiment of the invention, marigold flowers containing specific
percentage of xanthophyll content are selected and processed to get the pellets, which
10 are extracted using food grade solvents under specific conditions. The extract
containing about 14-17% by weight of lutein esters is obtained for further processing.
According to one more embodiment of the invention, fresh paprika fruit pods having
R,R zeaxanthin content of around 0.04—0.08% by weight are selected and dried to
15 specific moisture level and trans-R,R zeaxanthin content. Dried chili (paprika) are cut
in small pieces to obtain powder, which is then treated with food grade solvent/s to
get extract having 3-4% by weight of R,R zeaxanthin esters.
Marigold flowers (scientific name- Tagetes erecta, Family -Asteraceae): is a species
20 of the genus Tagetes native to Mexico. It is cultivated in Asia, Africa and South
America. Marigold is also cultivated in the districts of Karnataka, particularly in the
area of Hassan, Chickamangaluru, Mysore, Chamrajnagar, Davangere. For further
processing the Marigold flowers are delivered to plant situated at Hassan (Karnataka)
and Kangayam (Tamilnadu) India. The extraction is carried out by manual
25 intervention, in order to prepare a composition rich in xanthophyll, which selectively
contains trans-R,R zeaxanthin isomer along with tans-R,R lutein in specific weight
percentage, which is explicitly different from naturally existing composition of
marigold flowers and thus is enriched and safe for human consumption.
12
Another source used in the invention is Paprika (Capsicum annuum) which is a
species of the plant genus Capsicum (peppers) native to southern North America and
northern South America. This species is the most common and extensively cultivated
of the five domesticated capsicums. For the purpose of present invention, the raw
material may be either purchased from outside of India, or may 5 be cultivated as
Capsicum annuum in Karnataka.
The xanthophyll composition is comprised of at least 80% by weight of total
xanthophyll, of which at least 65% by weight being trans-R,R lutein and at least 10%
10 by weight being trans-R,R zeaxanthin.
According to a preferred embodiment the composition is substantially free of R,S and
S,S zeaxanthin .
15 In an embodiment, the xanthophyll composition is comprised of trans-R,R lutein and
trans-R,R zeaxanthin in the ratio of 4:1 to 6:1.
In an embodiment, the xanthophyll composition is comprised of trans-R,R lutein and
trans-R,R zeaxanthin in the ratio of 5:1.
In an embodiment, the xanthophyll composition is comprised of trans-R,R lutein 10
20 to 15 mg and trans-R,R zeaxanthin is 2 to 3 mg.
In an embodiment, the xanthophyll composition comprising trans-R,R lutein and
trans-R,R zeaxanthin is in the range of 0.1-10 micron.
In an embodiment, the obtained oil suspension composition as per the example 1
having particle size is in the range of 0.1-10 micron.
13
In an embodiment, the xanthophyll composition comprising increased bioavailability
of 1.5 – 4 folds against reference (Product B).
According to still one more embodiment of the invention, xanthophyll composition
described herein can be formulated using at least one pharmaceutical, nutraceutical or
food grade excipient or combination thereof. The excipient 5 selected from
bioavailability enhancing agent, carrier /vehicle, granulating agent, inert core, coating
agent, solvent, diluents, binder, lubricant, disintegrant, antioxidant, oil, surfactant,
solubilizer, emulsifier or any other excipient, which is known to a person skilled in
the art as excipient required for preparing palatable dosage form, acceptable to the
10 subject.
According to further embodiment of the present invention bioavailability enhancing
agent may be selected from nigella (black cumin oil), caraway (carum carvi) oil,
Phosphatidyl choline (lecithin), starch sodium octenyl succinate, Allspice Berry,
15 Amber Essence, Anise Seed, Arnica, Balsam of peru, Basil, Bay Leaf, Benzoin Gum,
Bergamot, Bois de Rose (Rosewood), Cajeput, Calendula (Marigold pot), White
Camphor, Caraway Seed, Cardamon, Carrot Seed, Cedarwood, Celery, German or
Hungarian Chamomile, Roman or English Chamomile, Cinnamon, Citronella, Clary
Sage, Clovebud, Coriander, Cumin, Cypress, Eucalyptus, Fennel, Siberian Fir needle,
20 Frankincense (Olibanum oil), Garlic, Rose Geranium, Ginger, Grapefruit, Hyssop,
Jasmine Absolute, Jojoba, Juniper Berry, Lavender, Lemon, Lemongrass, Lime,
Sweet Marjoram, Mugwort, Mullein Flower, Myrrh Gum, Bigarade Neroli, Nutmeg,
Bitter Orange, Sweet Orange, Oregano, Patchouly, Pennyroyal, Black Pepper,
Peppermint, Petitegrain, Pine Needle, Poke Root, Rose Absolute, Rosehip Seed,
25 Rosemary, Dalmation Sage, Sandalwood Oil, Sassafras, Spearmint, Spikenard,
Spruce (Hemlock), Tangerine, Tea Tree, Thuja (Cedar leaf), Thyme, Vanilla extract,
Vetivert, Wintergreen, Witch Hazel (Hamamelia) Extract, or Ylang (Cananga)
Extract or the combination thereof.
14
According to a preferred embodiment of the present invention bioavailability
enhancing agent selected from nigella (black cumin oil), caraway (carum carvi) oil,
Phosphatidyl choline (lecithin).
5
According to further embodiment the bioavailability enhancing agent present in range
of 1- 10% by weigh of composition.
According to further embodiment the bioavailability enhancing agent present in range
of 1- 5% by weigh of composition.
10 According to further embodiment the bioavailability enhancing agent present in range
of 1- 3% by weigh of composition.
According to a preferred embodiment of the present invention solubility enhancer
may selected from thyme oil, olive oil, linseed oil (flaxseed oil) or mixture thereof.
15
According to further embodiment the solubility enhancer present in range of 1- 10%
by weigh of composition.
According to further embodiment the solubility enhancer present in range of 1- 5% by
weigh of composition.
20
According to further embodiment of the present invention antioxidant may be
selected from but not limited to, but not limited to butylated hydroxytoluene,
butylated hydroxyanisole, ascorbic acid, ascorbyl palmitate, sodium ascorbate, mixed
tocopherol, alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol,
25 synthetic racemic tocopherols and tocopherol esters or the combination thereof.
According to further embodiment the range of the antioxidant is 1-10% by weight of
composition. According to further embodiment the antioxidant present in range of 1-
10% by weight of composition. According to further embodiment the antioxidant
15
present in range of 1- 5% by weight of composition. According to further
embodiment the antioxidant present in range of 1- 3% by weight of composition.
According to further embodiment of the present invention carrier/ vehicle may be
selected from but not limited to sunflower oil, coconut oil, corn oil, 5 cottonseed oil,
canola oil, olive oil, palm oil, peanut oil, safflower oil, sesame oil, soybean oil,
rapeseed oil and medium chain triglyceride (MCT) oil or the combination thereof.
According to further embodiment of the present invention Hydrophilic carrier may be
10 selected from but not limited to celluloses such as alkyl cellulose (methyl cellulose), a
hydroxyalkyl cellulose (e.g., hydroxymethyl cellulose, Hydroxypropyl cellulose),
carboxyalkyl cellulose (e.g., carboxymethyl cellulose and alkali metal salts thereof,
such as sodium salts), a carboxyalkylalkyl cellulose (e.g., carboxymethylethyl
cellulose), carboxyalkyl cellulose ester (e.g., carboxymethyl cellulose butyrate,
15 carboxymethyl cellulose propionate, carboxymethyl cellulose acetate butyrate and
carboxymethyl cellulose acetate propionate) or the combination thereof.
Polyacrylates such as polymethacrylate, a methacrylate copolymer (e.g., a
methacrylic acid-methyl methacrylate copolymer, dimethylaminoethyl methacrylatebutyl
methacrylate-methyl methacrylate copolymer, and a diethylaminoethyl
20 methacrylic acid-methyl methacrylate copolymer), and an ethacrylate copolymer (e.g.
methacrylic acid ethacrylate copolymer) or the combination thereof; povidones such
as polyvinyl pyrrolidone, polyvinyl acetate ester (e.g., polyvinyl acetate phthalate
(PVAP), and polyethylene glycol polyvinylacetate copolymer (e.g. polyethylene
glycol-polyvinylcaprolactam-polyvinylacetate copolymer) or the combination
25 thereof.
Starch and starch derivatives such as Modified Starch, corn starch, potato starch,
pregelatinized starch, dextrins, acid-treated starch, alkaline-treated starch, bleached
starch, oxidized starch, enzyme-treated, monostarch phosphate, distarch phosphate,
16
phosphateddistarch phosphate, acetylated distarch phosphate, starch acetate,
acetylated distarch adipate, hydroxypropyl starch, hydroxypropyl distarch phosphate,
hydroxypropyl distarch glycerol, starch sodium octenyl succinate, acetylated oxidized
starch and maltodextrin or the combination thereof.
5
Gums such as pectin alginate, carrageenan agar, gum arabic, gum tragacanth, gum
karaya, gum ghatti, gum guar, Locust bean gum, Tara gum, Xanthan gum, Gellan
gum and Welan gum or the combination thereof; sugars such as fructans, sucrose,
glucose and fructose or the combination thereof.
10
According to preferred embodiment the carrier is MCT oil. According to further
embodiment MCT oilis present in range of 20-80% by weight of composition.
According to further embodiment MCT oilis present in range of 50-70% by weight of
composition.
15
According to preferred embodiment the carrier is modified starch. According to
further embodiment starchis present in range of 10-40% by weight of composition.
According to further embodiment starchis present in range of 10-30% by weight of
composition. According to further embodiment starchis present in range of 10-20%
20 by weight of composition.
According to one embodiment, xanthophyll composition comprising trans enriched
trans-R,R isomers of lutein and zeaxanthin in specific ratio can be further formulated
in different forms like beadlets, powders, oil suspensions, granules, capsules, tablets,
25 films or any other suitable oral, parenteral or topical dosage forms using
pharmaceutically or nutraceutically acceptable excipients and/or carriers, but
preferably oil suspension and beadlets.
17
As per one embodiment of present invention, the process for manufacture of beadlets
is comprised of spraying carotenoid emulsion in a square chamber and embedding it
in cold matrix bed, wherein square chamber has provision for continuous rotation and
circulation of cold water through jacketed vessel.
5
As per one embodiment of the present invention, the process for preparation of
carotenoid emulsion is comprised of solvent phase and aqueous phase and mixing the
two phases under stirring at specific conditions to get emulsion. The solvent phase as
described herein is prepared by dissolving active in solvent phase comprised of
10 organic solvent, antioxidant and at least one more oily excipient, at specific
conditions and filtering out to remove any insoluble residue. The aqueous phase as
described herein is prepared by dissolving hydrophilic carrier and antioxidant in
water.
15 As per one more embodiment of this invention, the solvent phase is added to water
phase under stirring to get emulsion and homogenized in high pressure homogenizer.
This homogenized emulsion is then subjected to evaporation under vacuum to get
solvent free carotenoid emulsion.
20 As per one important embodiment of the present invention, the process for the
manufacture of carotenoid beadlets is comprised of spraying carotenoid emulsion in
square chamber, which is embedded and solidified in cold matrix bed to form the
beadlets.
25 As per one more embodiment of this invention, the process for the manufacture of
carotenoid beadlets is carried out in square chamber which has provision of two inlets
at top for spray nozzle of emulsion and for hydrophilic matrix. The square chamber
also has two outlets at bottom of chamber for collection of beadlets and unused
matrix bed.
18
As per one more important embodiment of this invention, the square chamber has
provision for continuous rotation as well as circulation of cold water through jacketed
vessel to maintain inside temperature below 20°C.
5
As per the main embodiment of the present invention, the process for manufacture of
carotenoid beadlets is comprised of loading the matrix bed through one inlet at the
top of square chamber and maintaining it cold with the help of circulating cold water.
The emulsion is sprayed through spray nozzle of rotating square chamber and the
10 droplets are embedded in particles of cold matrix bed to form beadlets.
The beadlets formed by the above process are collected through outlet at the bottom
of square chamber and subjected to drying until desired moisture level is attained.
15 As per one more embodiment of the present invention, matrix bed which is loaded in
square spray chamber is selected from, but not limited to starch derivatives such as
wax modified maize starch, starches and modified starches in combination with talc,
silicic acid, hydrogenated fats, metal salt of higher fatty acid such as calcium stearate,
aluminium silicate of alkali metal/ alkaline earth metal,sodium or calcium aluminium
20 silicate, alumina, calcium silicate, powdered silica gel, magnesium carbonate and
magnesium oxide or the combination thereof.
As per a preferred embodiment of the present invention, the process for preparation
of xanthophyll composition in the oil suspension form comprised of obtaining trans-
25 R,R lutein and trans-R,R zeaxanthin with selective ratio of 4:1 to 6:1 by extracting
from marigold and paprika oleoresin. The marigold and paprika extracted to get trans-
R,R lutein and trans-R,R zeaxanthin called as concentrate which further undergoes
for micronization by using air jet mill to obtain particle size in the range of 0.1 -10
micron. The micronized trans-R,R lutein and trans-R,R zeaxanthin with selective
19
ratio with preferred particle size obtained is mixed by adding bioavailability
enhancing agent, solubility enhancer, antioxidant, carrier oil and optionally flavor
using ball mill at room temperature. The obtained oil suspension composition has
particle size in the range of 0.1 -10 micron.
5
The particle size of the trans-R,R lutein and trans-R,R zeaxanthin is in the range of
0.1-10 micron.
The other excipient such as carrier oil/vehicle, bioavailability enhancing agent,
10 solubility enhancer, stabilizer, and antioxidant is added in ball mill.
The particle size of obtained oil suspension composition as per the example 1 having
particle size is in the range of 0.1-10 micron.
15 As per the further embodiment, the Xanthophyll composition in the form of oil
suspension comprising Xanthophyll and pharmaceutically and/or nutraceutically
acceptable excipient shaving enhanced bioavailability.
At the outset of the description that follows, it is to be understood that the ensuing
20 description only illustrates a particular form of this invention. However, such a
particular form is only an exemplary embodiment and is not intended to be taken
restrictively to imply any limitation on the scope of the present invention.
Example 01:
25 According to preferred embodiments we have performed different size batches from
small to large scale to support the industrial viability in process.
Oil suspension composition preparation; Batch size 500 g
20
Batch size 500g
Sr. No Name of the Ingredient Standard
Quantity (g)
%W/W
1 Micronized trans-R,R lutein
and trans-R,R zeaxanthin
170 35
2 Olive Oil USP 25 5
3 MCT Oil 260 51
4 Mixed Tocopherol 70%-SF 10 2
5 Thyme Oil 5 1
6 Linseed oil (flaxseed oil) 10 2
7 D-Limonene 10 2
8 Lecithin
(Phosphatidylcholine)
10 2
A weight quantity of marigold oleoresin (33.5 kg) containing 16.6% xanthophyll
content (by spectrophotometric method) and 13.6 lutein content (by HPLC method).
Paprika oleoresin (16.5 kg) containing 5.7% xanthophyll content (by
spectrophotometric method) and 3.5% zeaxanthin content 5 (by HPLC method).
Each oleoresin was added in reactor with aqueous alcoholic alkali solution (10 kg
potassium hydroxide in 15 ltr water and 62.5 ltr of ethanol). The mixture was heated
in reactor with stirring at 80ºC for a period of 4 hours. The degree of hydrolysis was
monitored by HPLC during the reaction and reaction terminated by adding water 250
10 ltr. The homogenize the reaction mass at room temperature and add ethyl acetate (300
ltr) in reactor for liquid- liquid extraction and repeate the process for 3-5 times. The
EA layer separated out and washed with water (1:1) at room temperature. The water
washed ethyl acetate further taken for concentration and to get concentrated material.
The concentrated material isolated by using hexane to the removing impurities and to
15 get semi-purified crystals (22.5 kg) of xanthophyll composition. The semi-purified
21
crystals obtained were purified with ethanol 5 volumes at room temperature, followed
by filtration (7.8 kg). The resulting crystals were vacuum dried at temperature of
about 50 to 55°C for 36 hrs. The yield of the dried xanthophyll crystals was 9.6 kg
(9.6%). The xanthophyll content was 87.2% by weight (UV/Vis-spectrophotometry)
out of which the contents of trans-R,R lutein and trans-R,R zeaxanthin 5 were 70.3%,
and 14.4% and area% of trans-R,R lutein and trans-R,R zeaxanthin were 80.6%, and
16.5%respectively as determined by normal phase HPLC analysis.
The obtained concentrate (2.5 kg) containing 87.2% xanthophyll content (by
10 spectrophotometric method) was taken for Micronization. The Micronization of
marigold paprika extract concentrate containing particle size (DV90) 1690µm was
micronized by using air jet mill. The resulting micronized crystals (2.2kg) containing
particle size (DV90) 3.24 µm (figure 8) and the xanthophyll content was 89.09% by
weight (UV/Vis-spectrophotometry) out of which the contents of trans-R,R lutein and
15 trans-R,R zeaxanthin were 71.62%, and 14.75% and area% of trans-R,R lutein and
trans-R,R zeaxanthin were 80.39%, and 16.56% respectively as determined by
normal phase HPLC analysis.
The micronized marigold paprika extract concentrate (170 g) containing 87.2%
20 xanthophyll content (by spectrophotometric & HPLC method) was used to make
formulation as oil suspensions. The micronized marigold paprika extract concentrate
was mixed by adding oil (260g) medium Chain triglycerides (MCT) with 14% (70 g)
other ingredients as olive oil, thyme oil, linseed oil (flaxseed oil), D-Limonene,
Lecithin (Phosphatidylcholine) (Obtained from Sunflower), mixed tocopherol by
25 using ball mill at room temperature. The mixture was sieved and unloaded (500g)
analyzed for containing particle size (DV90) 2.66 µm (Figure 7) and xanthophyll
content was 28.05% by weight (UV/Vis-spectrophotometry) out of which the
contents of trans-R,R lutein and trans-R,R zeaxanthin were 22.64%, and 4.63% and
22
area% of trans-R,R lutein and trans-R,R zeaxanthin were 80.72%, and 16.49%
respectively as determined by normal phase HPLC analysis.
Example 02:
Oil suspension preparation; 5 Batch size 18 Kg
Batch size 18.0kg
Sr. No Name of the Ingredient Standard
Quantity (kg)
%W/W
1 Micronized trans-R,R lutein and
trans-R,R zeaxanthin
6.28 35
2 Olive Oil USP 0.9 5
3 MCT Oil 9.2 51
4 Mixed Tocopherol 70%-SF 0.36 2
5 Thyme Oil 0.18 1
6 Linseed oil (flaxseed oil) 0.36 2
7 D-Limonene 0.36 2
8 Lecithin (Phosphatidylcholine) 0.36 2
A weighted quantity of micronized marigold paprika extract concentrate (6.28 kg)
containing 80.97% xanthophyll content (by spectrophotometric & HPLC method)
10 was used to make formulation as oil suspensions. The micronized marigold paprika
extract concentrate was mixed by adding oil (9.2kg) medium Chain triglycerides
(MCT) with 14% (2.52 kg) other ingredients as olive oil, thyme oil, linseed oil
(flaxseed oil), D-Limonene, Lecithin (Phosphatidylcholine) (Obtained from
Sunflower), mixed tocopherol by using ball mill at room temperature. The mixture
15 was sieved and unloaded (18kg) analyzed for containing particle size (DV90) 2.62µm
23
and xanthophyll content was 28.23% by weight (UV/Vis-spectrophotometry) out of
which the contents of trans-R,R lutein and trans-R,R zeaxanthin were 22.6%, and
4.71% and area% of trans-R,R lutein and trans-R,R zeaxanthin were 80.16%, and
16.68% respectively as determined by normal phase HPLC analysis.
5
Example 03:
Beadlet preparation (15 %); Batch Size 24.2 kg
Stages Ingredients (% w/w)
Cake
Phase
Micronized trans-R,R
lutein and trans-R,R
zeaxanthin
24.2
Dichloromethane
(MDC)
895
Tocopherol 2.42
Water Phase
Modified starch 53.8
Sodium Ascorbate 2.5
Water 274.0
Powder catch Starch N 200 17.5
Batch Size 100
Set assay 17.0
% Solid in ATFE Solution 23.18
Total Solution quantity 1251.0
Procedure:
10
A. A micronized marigold paprika extract concentrate was dissolved into
methylene dichloride followed by addition of Tocopherol under stirring. The
solution was warmed for 30 min at 40-45°C till it becomes clear and further
24
cooled to room temperature. The solution was filtered to get clear brownish
colored solution.
B. Modified starch was dissolved in water and warmed up to 50-55°C, till it
becomes clear solution under stirring. Cool the aqueous system to
5 roomtemperature.
C. Stage A solution was mixed into stage B solution under stirring and sodium
ascorbate was added to the mixture.
D. Stage C solution was homogenized at 15,000-20,000 rpm for 5 min interval.
E. Stage D solution was added into rotary evaporator and methylene dichloride
10 was removed at 60°C in the water bath with rotation at 4 rpm under vacuum
and cooled it up to room temperature.
F. 3.5 kg Starch initially dried at 105°C for 4 hours was added into spray pan and
dry ice was added to the chamber to bring the temperature down to 2°C.
Spraying of stage E solution was started onto the dry starch under fluidization.
15 G. Coated macular carotenoid beadlets were collected by sifting stage F material
and dried the beadlets into tray oven at 35°C - 40°C and sifted the dried
beadlets through mesh 60 & 40 sieves.
Example 04:
20 Beadlet preparation (20%); Batch Size 31.7 kg
Stages Ingredients Beadlets 20% w/w
Cake
Phase
Micronized trans-R,R
lutein and trans-R,R
zeaxanthin
31.7
Dichloromethane 1172
Tocopherol 3.2
Water Phase Modified starch 45.2
25
Sodium Ascorbate 2.5
Water 274
Powder catch Starch N 200 17.5
Batch Size 100
Set assay 22
% Solid in ATFE Solution 23.18
Total Solution quantity 1528
Procedure:
A. Micronized marigold paprika extract concentrate was dissolved into
dichloromethane followed by addition of Tocopherol under stirring.
The solution was warmed for 30 min at 40-45°C till it becomes 5 clear and further
cooled to room temperature. The solution was filtered to get clear brownish colored
solution.
B. Modified starch were dissolved in water and warmed up to 50-55°C, till it becomes
clear solution under stirring. Cooled the aqueous system to room temperature.
10 C. Stage A solution was mixed into stage B solution under stirring and sodium
ascorbate was added to the mixture.
D. Stage C solution was homogenized at 15,000-20,000 rpm for 5 minutes interval.
E. Stage D solution was added into rotary evaporator and methylene dichloride was
removed at 60°C in the water bath with rotation at 4 rpm under vacuum and cooled it
15 up to room temperature.
F. 3.5 kg Starch initially dried at 105°C for 4 hours was added into spray pan and dry
ice was added to the chamber to bring the temperature down to 2°C. Spraying of stage
E solution was started onto the dry starch under fluidization.
G. Coated macular carotenoid beadlets were collected by sifting stage F material and
20 dried the beadlets into tray oven at 35°C - 40°C and sifted the dried beadlets through
mesh 60 & 40 sieves.
26
Example 05:
Beadlet preparation (10 %); Batch Size 19.5 kg
Beadlets 10% w/w
Stages Ingredients % W/W
Water
Phase
Micronized trans-R,R lutein and
trans-R,R zeaxanthin
19.5
Tocopherol 3.5
Modified starch 54.8
Sodium ascorbate 2.2
Water 132
Powder
catch
Starch N 200 20.0
Batch Size 100.0
Set assay 13.0
% Solid in Solution 40.00
Total Solution quantity 1000.0
5
Procedure:
A. Disperse the Modified starch& Sodium ascorbate binder into warm water (50-
55°C) under stirring to from the clear solution.
10 B. Added Marigold Paprika extract Conc-Micronized and Tocopherol into Stage
A dispersion under stirring to form the solution for 15 min.
C. StageB solution was processed under polytron high shear stirrer for uniform
mixing of the cake into binder solution at 5000 to 12000 RPM for 15 min,
observed the Temp of the solution and flow ability.
27
D. Added the Stage C solution into horizontal grinding mill inlet and processed it
for repeated passes for size reduction and proper emulsification of solution
using following process parameters.
E. After ATFE pass, Catch the solution on fluidized starch N 200 at desired
process 5 parameters.
F. Dry the beadlets in oven at 55°C to achieve the LOD below 5%
The Bioavailability study design for the xanthophyll composition comprising
trans-R,R lutein and trans-R,R zeaxanthin in form of oil suspension was carried
10 out as Study 1 below:
Study 1:
For ease of understanding, sample nomenclature for Product A is Micro OS R+ and
Product B is Reference sample. In addition, in examples wherever lutein and
15 zeaxanthin are mentioned, it should be considered as trans-R,R lutein and trans-R,R
zeaxanthin.
Objective:
20 To compare the bioavailability of trans-R,R lutein 10 mg and trans-R,R zeaxanthin 2
mg from a single oral dose (oil suspension) as a Product A (Micro OS R +) with
product B (Reference) formulation (trans-R,R lutein 10 mg and trans-R,R zeaxanthin
2 mg per capsule) supplementation in healthy, adult human subjects under fed
conditions.. Product B is a marketed product Florglo® (trans-R,R lutein) and
25 OptisharpTM (trans-R,R zeaxanthin).
Study Design: Double blind, balanced, randomized, two-treatment, single-period,
single-dose, parallel study
Preparation:
28
Micro OS R+ sample: prepared as per example 01 in 5:1 ratio
Product B: a marketed product Florglo and Optisharp (FloraGLO® Lutein 20 % SAF
and OPTISHARP™ Natural (Zeaxanthin) 14% Safflower Oil were calculated and
mixed to maintain the 5:1 ratio wherein trans-R,R lutein 10 mg and trans-R,R
zeaxanthin 5 2 mg per capsule).
Number of volunteers: 90 healthy volunteers (45 volunteers per group)
Housing of volunteers: 7 days housing pre and post study to control for diet with
minimal amount of lutein and zeaxanthin
Pharmacokinetic (PK) Assessments:
10
Baseline corrected Serum concentration of trans-R,R lutein and trans-R,R zeaxanthin
were measured at different time points:
Time points (hours): -48, -24, 0, 2, 4, 6, 8, 10, 12, 16, 20, 24, 48 and 72 hours
15 ? Pharmacokinetic parameters (Ln(Cmax), Ln(AUC0-72),Ln(AUC0-T and
Tmax))
? 90% CI, T/R ratio
Bioanalytical Procedure:
20 Lutein and Zeaxanthin (lutein comprising trans-R,R lutein and zeaxanthin comprising
trans-R,R zeaxanthin) was estimated in serum using a validated HPLC method.
RESULTS FOR TRANS-R,R LUTEIN AND TRANS-R,R ZEAXANTHIN
25
Baseline Corrected Serum trans-R,R lutein Concentrations for Product A
(Micro OS R+) and Product B (Reference)
29
Time
Points
(hrs.)
Product A
Micro OS R+
(N=45)
Product B
Reference
(N=45)
Arithmetic Mean SD
Arithmetic
Mean
SD
ANOVA
P-Value
0.00 11.8829 33.66531 4.9148 13.77006 0.2021
2.00 14.4350 36.03639 2.4143 6.83637 0.0305*
4.00 33.7202 48.52837 9.0962 15.06611 0.0016*
6.00 89.4877 103.83044 32.2815 31.06571 0.0006*
8.00 138.4649 121.92331 55.1417 52.78092 <.0001*
10.00 147.0603 103.61156 70.7683 59.17610 <.0001*
12.00 180.3070 124.17764 83.4900 64.75716 <.0001*
16.00 192.2944 144.60898 80.1977 68.75500 <.0001*
20.00 207.8854 168.85120 70.0933 55.60889 <.0001*
24.00 190.2843 152.47004 74.3409 66.74332 <.0001*
48.00 167.3378 178.20127 50.8520 44.00988 <.0001*
72.00 132.5861 115.34769 46.2418 48.66596 <.0001*
* P value <0.05
? Baseline corrected serum trans-R,R lutein concentrations for Micro OS R+ group
were significantly higher (P<0.05) compared to Reference group 5 at all-time points
(2 to 72 hrs) post dose. (Fig 1)
? Baseline corrected serum trans-R,R lutein concentrations for Micro OS R+ group
showed more than 2X folds higher absorption at all time points and 3X folds
higher absorption at Tmax (20 hrs) compared to Reference group.
10
30
Baseline Corrected Pharmacokinetic Parameters for Trans-R,R Lutein
Parameters
Geometric Least Square
Mean A/B Ratio
(%)
P-Value
Log
Transformed
Product A
Micro OS R+
(N=45)
Product B
Reference
(N=45)
90% Confidence
Interval (%)
Ln(Cmax)
(ng/ml) 219.1539 87.0140 251.86 191.97 330.44
<.0001*
Ln(AUC0-72)
(hr*ng/ml) 8499.8925 2892.0893 293.90 217.33
397.46
<.0001*
Ln(AUC0-T)
(hr*ng/ml) 8264.0226 2622.9697 315.06 224.50
442.17
<.0001*
Tmax (hours)$
20.00 (4.00-
72.00)
16.00 (6.00 –
48.00)
-
- -
0.0564
$ Median (Minimum-Maximum)
5 * P value <0.05
? Micro OS R+ group showed significantly higher absorption for trans-R,R
lutein compared to Reference group as evident from the significantly higher
(P<0.05) baseline corrected Log transformed Cmax, AUC0-72 and AUC0-T.
10 ? Micro OS R+ group reached highest serum trans-R,R lutein concentration
(Tmax) at 20 hrs whereas Reference group achieved highest serum trans-R,R
lutein concentration at 16 hrs.
? 90% confidence interval for Cmax and AUC parameters were significantly
higher than the 80-125% bioequivalence criteria demonstrating the
15 significantly superior bioavailability of Micro OS R+ group over Reference
group.
Folds of Test compared to Reference for Baseline Corrected trans-R,R Lutein
20 PK Parameters.
31
Parameters
Geometric Least Square Mean Folds of Test
compared to
Reference
Product A
Micro OS R+
Product B
Reference
Ln(Cmax)
(ng/ml) 219.1539 87.0140 2.5X
Ln(AUC0-72)
(hr*ng/ml) 8499.8925 2892.0893 2.9X
Ln(AUC0-T)
(hr*ng/ml) 8264.0226 2622.9697 3.2X
• Micro OS R+ group showed significantly higher absorption for trans-R,R
lutein compared to Reference group as evident from the 2.5X folds Cmax,
2.9X folds AUC0-72 and 3.2X folds AUC0-T levels 5 (Fig 2 and Fig 3).
RESULTS FOR TRANS-R,R ZEAXANTHIN
Baseline Corrected Serum trans-R,R Zeaxanthin Concentrations for Product A
10 (Micro OS R+) and Product B (Reference)
Time
Points
(hrs)
Product A
Micro OS R+
(N=45)
Product B
Reference
(N=45)
Arithmetic
Mean SD Arithmetic
Mean SD ANOVA PValue
0.00 9.4402 12.51823 6.2203 7.61907 0.1441
2.00 12.9155 19.38294 6.4335 11.56039 0.0572
4.00 20.5625 24.90861 10.8184 13.56510 0.0235*
6.00 32.1562 34.49185 15.4633 15.08300 0.0038*
8.00 34.0454 26.58597 18.6964 21.76251 0.0035*
10.00 31.2432 18.45573 16.8040 20.00775 0.0006*
12.00 33.2990 20.60122 22.8680 23.66415 0.0283*
32
16.00 30.6575 25.63941 15.1527 21.75149 0.0027*
20.00 32.3948 27.30477 11.3353 13.93915 <.0001*
24.00 29.8132 23.06656 16.8866 20.41164 0.0060*
48.00 38.9101 38.84572 18.0805 20.09565 0.0019*
72.00 38.3075 31.39057 23.5634 25.88299 0.0171*
* P value <0.05
• Baseline corrected serum trans-R,R zeaxanthin concentrations 5 for Micro OS
R+ group were significantly higher (P<0.05) compared to Reference group
between 4 to72 hrs time points post dose. (Fig 4)
• Baseline corrected serum trans-R,R zeaxanthin concentrations for Micro OS
R+ group showed more than 1.5X folds higher absorption at all time points
10 and 2.9X folds higher absorption at Tmax (20 hrs) compared to Reference
group
Baseline Corrected Pharmacokinetic Parameters for trans-R,R zeaxanthin
Parameters
Geometric Least
Square Mean
A/B
Ratio
(%)
90%
Confidence
Interval (%) P_Value
Log
Transformed
Product A
Micro OS
R+
(N=45)
Product B
Reference
(N=45)
Ln(Cmax)
(ng/ml) 56.9452 32.5522 174.93
140.70
217.50
<.0001*
Ln(AUC0-
72)
(hr*ng/ml)
1929.9318 861.6456 223.98
159.64
314.26
0.0002*
33
Ln(AUC0-
T)
(hr*ng/ml)
1817.2934 814.1689 223.21
154.35
322.79
0.0005*
Tmax
(hours)$
20.00
(0.00-
72.00)
24.00
(0.00-
72.00)
-
- -
0.7866
$ Median (Minimum - Maximum)
* P value <0.05
• Micro OS R+ group showed significantly higher absorption for trans-R,R
zeaxanthin compared to Reference group as evident from 5 the significantly
higher (P<0.05) baseline corrected Log transformed Cmax, AUC0-72 and
AUC0-T.
• Micro OS R+ group reached highest serum trans-R,R zeaxanthin
10 concentration (Tmax) at 20 hrs whereas Reference group achieved highest
serum concentration at 24 hrs.
• 90% confidence interval for Cmax and all AUC parameters were significantly
higher than the 80-125% bioequivalence criteria demonstrating the
significantly superior bioavailability of trans-R,R zeaxanthin from Micro OS
15 R+ group over Reference group.
Folds of Test compared to Reference for Baseline Corrected trans-R,R
zeaxanthin PK Parameters.
Parameters
Geometric Least Square Mean
Folds of Test compared
Product A to Reference
Micro OS R+
Product B
Reference
Ln(Cmax)
(ng/ml) 56.9452 32.5522 1.7X
Ln(AUC0-
72) 1929.9318 861.6456 2.2X
34
(hr*ng/ml)
Ln(AUC0-T)
(hr*ng/ml) 1817.2934 814.1689 2.2X
• Micro OS R+ group showed significantly higher absorption for trans-R,R
zeaxanthin compared to Reference group as evident from the 1.7X folds
Cmax and 2.2X folds AUC0-72 & AUC0-T levels
5
Conclusion of study on Oil suspension (Micro OS R+; Product A) composition:
Micro OS R+ (Product A) composition showed significantly higher absorption for
trans-R,R Lutein in the range of 2.9 to 3.2 folds and trans-R,R zeaxanthin in 2.2
folds when compared to Reference (Product B) composition in terms of achieving
10 higher serum concentration at multiple timepoints measured post dose; higher
absorption, 90% confidence interval for Cmax and all AUC parameters were
significantly higher than the 80-125% bioequivalence criteria demonstrating the
significantly superior bioavailability of Micro OS R+ group over Reference group.
15 ABBREVIATIONS:
Terms Meaning
Cmax Highest concentration of an investigational product in the serum
AUC0-72 Area under the serum concentration versus time curve, from time 0 to 72
hours
AUC0-t Area under the serum concentration versus time curve, from time 0 to last
measurable concentration
Tmax Amount of time that an investigational product is present at the maximum
concentration in serum
35

15
20 ,CLAIMS:WE CLAIM:
1. A xanthophyll composition comprising:
a) trans-R,R lutein in the range of 65-85% by weight;
b) trans-R,R zeaxanthin in the range of 10-30% by weight;
c) one or more carrier and/or vehicle in the range of 5 20-80% by weight;
d) one or more solubility enhancer in the range of 1-10% by weight;
e) optionally one or more flavoring agent in the range of 1-5 % by weight;
f) one or more bioavailability enhancing agent in the range of 1-10% by
weight; and
10 g) one or more antioxidant agent in the range of 1-10% by weight.
Wherein trans-R,R lutein and trans-R,R zeaxanthin extracted from marigold
and paprika oleoresins and are present in the ratio of 4:1 to 6:1 and the said
composition is substantially free from R,S zeaxanthin and S,S zeaxanthin.
15 2. The xanthophyll composition as claimed in claim 1, wherein the particle size
of trans-R,R lutein and trans-R,R zeaxanthin are in the range of 0.1-10 micron.
3. The xanthophyll composition as claimed in claim 1, wherein carrier and/or
vehicle is selected from medium chain triglycerides (MCT) oil, sunflower oil,
20 coconut oil, corn oil, cottonseed oil, canola oil, palm oil, peanut oil, safflower
oil, sesame oil, soybean oil, rapeseed oil and medium chain triglyceride or the
combination thereof, preferably MCT oil.
4. The xanthophyll composition as claimed in claim 1, wherein the solubility
25 enhancer is selected from thyme oil, olive oil, linseed oil (flaxseed oil) or
mixture thereof.
36
5. The xanthophyll composition as claimed in claim 1, wherein the bioavailability
enhancing agent is selected from nigella (black cumin oil), caraway (carum
carvi) oil, Phosphatidyl choline (lecithin), or mixture thereof.
6. The xanthophyll composition as claimed in claim 1, wherein 5 an antioxidant is
selected from tocopherol and Sodium ascorbate or mixture thereof.
7. The xanthophyll composition as claimed in claim 1, wherein the particle size
of obtained composition is in the range of 0.1-10 micron.
10
8. The xanthophyll composition as claimed in claim 1, wherein the composition
has increased bioavailability of 1.5 –4 folds against reference.
9. The xanthophyll composition as claimed in claim 1, wherein the composition
15 has increased concentrations in serum in 2-4 times against reference.
10. The xanthophyll composition as claimed in claim 1, wherein the formulation is
in the form of beadlets, powders, oil suspension, granules, capsules, tablets,
films.
20
11. The xanthophyll composition as claimed in claim 1, wherein the composition
is in the form oil suspension.
12. The xanthophyll composition as claimed in claim 10, wherein the oil
25 suspension is prepared by comprising steps of:
a) trans-R,R lutein and trans-R,R zeaxanthin is obtained from marigold
and paprika oleoresinsas claimed in claim 1,with selective ratio of 4:1
to 6:1;
37
b) micronize trans-R,R lutein and trans-R,R zeaxanthin from step (a), by
using air jet mill to obtained particle size in the range of 0.1 -10
micron;
c) micronized trans-R,R lutein and trans-R,R zeaxanthin obtained in step
(b) is mixed with carrier 5 and /or vehicle;
d) add solubility enhancer to step (c) with flavoring agent;
e) add bioavailability enhancing agent and antioxidant agent to step (d);
f) mix the suspension obtained in step (e) using ball mill to get
homogenous oil suspension; and
10 g) sieve the mixture obtained from step (f) to get resultant particle size of
oil suspension.
13. The xanthophyll composition as claimed in claim 10, wherein the beadlets are
prepared by comprising steps of:
15 a) trans-R,R lutein and trans-R,R zeaxanthin is obtained from marigold
and paprika oleoresinsas claimed in claim 1,with selective ratio of 4:1
to 6:1;
b) micronize trans-R,R lutein and trans-R,R zeaxanthin from step (a), by
using air jet mill to obtained particle size in the range of 0.1 -10
20 micron;
c) dissolve micronized trans-R,R lutein and trans-R,R zeaxanthin in
methylene dichloride followed by antioxidantand optionally
bioavailability enhancing agent; andwarm followed by cool the
solutionand filtered to get clear brownish colored solution;
25 d) dissolve modified starch in water and warm till it becomes clear
solution andcool to room temperature;
e) mix Stage (c) solution into stage (d) solution under stirring and add
sodium ascorbate to the mixture under homogenization;
38
f) add Step (e)solution into rotary evaporator to remove methylene
dichloride under vacuum and cool;
g) spray of step (f) solution onto the dry starch under fluidization; and
h) collect coated macular carotenoid beadlets and dry.
5
14. The xanthophyll composition as claimed in claim 10, wherein the beadlets are
prepared by comprising steps of:
a) trans-R,R lutein and trans-R,R zeaxanthin is obtained from marigold
and paprika oleoresinas claimed in claim 1,with selective ratio of 4:1
10 to 6:1;
b) micronize trans-R,R lutein and trans-R,R zeaxanthin from step (a), by
using air jet mill to obtained particle size in the range of 0.1 -10
micron;
c) disperse the modified starch & sodium ascorbate into warm water to
15 from the clear solution;
d) add micronized tocopherol into Step (c) dispersion; and
e) process under polytron high shear stirrer for uniform mixing andpass
through horizontal grinding mill for size reduction.
15. The xanthophyll composition as claimed in claim 1, 12(a), 13(a) and 14 (a),
wherein the ratio of trans-R,R lutein and trans-R,R zeaxanthin is 5:1.