Xanthophyll Compositions Containing Lutein and Zeaxanthin with Enhanced Bioavailability - Patent application

Abstract

The present invention relates to a xanthophyll composition comprising selected isomers of trans-R,R lutein and trans-R,R zeaxanthin in a selected ratio and with a preferred particle size, and pharmaceutically and / or nutraceutical acceptable excipients, such as carriers, solubility enhancers, bioavailability enhancers, antioxidants, and optionally flavoring agents, thereby enhancing bioavailability. The xanthophyll composition is at least 80% total xanthophylls, and contains at least 65% by weight trans-R,R lutein and at least 10% by weight trans-R,R zeaxanthin, prepared by combining marigold and paprika oleoresins in a proportion to provide a preferred ratio of trans-R,R lutein and trans-R,R zeaxanthin.

Description

[Technical field]

[0001] The present invention relates primarily to xanthophyll compositions comprising selective isomers of lutein and zeaxanthin, pharma- ceutically and / or nutraceutical acceptable excipients and solubility enhancers, antioxidants, together with bio-enhancing agents that enhance the bioavailability of the active substances. The present invention particularly relates to xanthophyll compositions comprising selective trans-R,R lutein and trans-R,R zeaxanthin, and at least one bioavailability enhancer, solubility enhancer, and antioxidant. The present invention further relates to xanthophyll compositions comprising more than 80% of the total xanthophyll, the composition being composed of selective isomers such as trans-R,R lutein and trans-R,R zeaxanthin. The present invention more specifically relates to a xanthophyll composition comprising 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 extraction process from two different plant sources using industrially viable process. The present invention further relates to enhancing the bioavailability of a xanthophyll composition comprising trans-R,R lutein and trans-R,R zeaxanthin by a combination of a micronization process to obtain a selective range of particle sizes in a preferred oil as a vehicle / carrier and selective solubility enhancer, antioxidant and bioenhancer. This surprisingly improved bioavailability is due to the combined effect of a micronization process to obtain a selective range of particle sizes in a preferred oil as a vehicle and selective bioenhancer including solubility enhancer and antioxidant that surprisingly exerts improved bioavailability. The present invention further relates to a xanthophyll composition comprising trans-R,R isomers of lutein and zeaxanthin in a specific ratio, which can be further formulated in different forms such as beadlets, powders, oily suspensions, granules, capsules, tablets, films or any other suitable oral, parenteral or topical dosage forms using pharma- ceutically or nutritionally acceptable excipients or carriers or mixtures thereof.The present invention further relates to xanthophyll compositions that are substantially free of R,S zeaxanthin and suitable for human consumption for use in a variety of eye and brain health applications. [Background technology]

[0002] Natural xanthophylls are abundantly present in and derived from various plant and animal sources. The beneficial health effects of natural xanthophylls and their antioxidant properties have been well documented in the scientific literature. The only obstacle to the effective utilization of these properties by the body is the low bioavailability and highly oxidizable nature of these nutrients. Understanding the long-felt need for bioavailability helps optimize the dosage of dietary supplements and drugs.

[0003] The oral bioavailability of many xanthophyll nutrients is limited due to low solubility in gastrointestinal fluids; molecular structure; metabolic enzymes that cause chemical transformations in the gastrointestinal tract; physicochemical and physiological processes such as low epithelial cell permeability, membrane transporters, and interactions with intestinal flora. The bioavailability of these agents can be improved by specially designing compositions that control their release, solubilization, transport, metabolism, and absorption in the gastrointestinal tract. Various approaches have been used to address these issues of low bioavailability of macular nutrients for use in the form of dietary supplements for human or veterinary use, but there is a strong need to improve bioavailability. The approach includes compositions containing specific weight percentages of trans-R,R zeaxanthin and trans-R,R lutein prepared from extracts of two different plant sources using food grade solvents and industrially viable processes of extraction, treatment with alkali, neutralization and purification, further formulated with selective particle size by micronization, preferred carriers or vehicles and bioavailability enhancers, which provide a combinatorial effect resulting in enhanced bioavailability.

[0004] US Patent No. 6,075,058(A) teaches a composition for enhancing the bioavailability of carotenoids. The US Patent teaches a mixture of 200 micrograms of the carotenoid lutein and 125 micrograms of the carotenoid zeaxanthin, 10 mg of cholesterol, 100 mg of olive oil, 20 mg of egg yolk phospholipids, 250 micrograms of alpha tocopherol, and 0.375 ml of 0.15 M aqueous sodium chloride solution.

[0005] WO1999 / 047001 relates to a method for increasing the absorption of carotenoids, more specifically to a method for increasing the absorption and bioavailability of lutein and zeaxanthin in humans and poultry by the use of lysolecithin and lecithin.

[0006] IN201821036199 relates to a purified xanthophyll composition comprising (trans R-R)-lutein and (trans R,R)-zeaxanthin, which accounts for more than 80% of the total xanthophylls, and a method for preparing them. The purified xanthophyll composition is composed of at least 85% by weight of (trans R,R)-lutein and at least 15% by weight of selective isomers such as (trans R,R)-zeaxanthin. The composition is prepared by a process of extraction. This is an industrially viable process, and purification is carried out by using food grade polar and non-polar solvents. The present invention mainly describes a process for preparing the composition, and does not fully mention the formulation composition.

[0007] The inventors of the present invention have conducted rigorous experiments to prepare a xanthophyll composition containing selective isomers of trans-R,R zeaxanthin and selective isomers of trans-R,R lutein. The composition is prepared by extracting two different plant sources and treating the xanthophyll ester-rich extract with alcoholic alkali. The resulting reaction mixture is then subjected to an extraction, isolation, and purification process using food-grade polar and non-polar solvents to selectively obtain a xanthophyll composition containing at least 15% by weight of the trans-R,R zeaxanthin isomer in combination with at least 80% by weight of trans-R,R lutein.

[0008] The present inventors have found that the enhanced bioavailability of xanthophyll compositions containing trans-R,R lutein and trans-R,R zeaxanthin is due to the combined effect of a micronization process to obtain a select range of particle sizes in a preferred oil as a vehicle, and selective bio-enhancements, solubility enhancers and antioxidants that exert a surprising improvement in bioavailability.

[0009] Objective of the invention The primary objective of the present invention is to provide a xanthophyll composition selectively containing trans-R,R lutein along with trans-R,R zeaxanthin in specific weight percentages prepared from extracts of two different plant sources using food grade solvents and industrially viable processes, and formulated with pharma- ceutical and / or dietary supplement acceptable excipients to enhance bioavailability.

[0010] A further primary objective of the present invention is to enhance the bioavailability of xanthophyll compositions including trans-R,R lutein and trans-R,R zeaxanthin through the combined effect of micronization process to obtain a select range of particle sizes in a preferred oil as a vehicle and selective bio-enhancements, solubility enhancers and antioxidants which exert surprising improvement in bioavailability.

[0011] A further object of the present invention is to provide a xanthophyll composition comprising trans-R,R isomer of lutein and trans-R,R isomer of zeaxanthin in a specific ratio, which is further formulated in different forms such as beadlets, powders, oil suspensions, granules, capsules, tablets, films, or any other suitable oral, parenteral or topical dosage forms using pharma- ceutically or nutritionally acceptable excipients and / or carriers.

[0012] Another object of the present invention is to provide a xanthophyll composition comprising trans-R,R zeaxanthin and trans-R,R lutein and a selective bioenhancer, which shows enhanced bioavailability with a more statistically significant 90% confidence interval (i.e., higher than the 80-125% bioequivalence criteria) for the test formulation, which shows superior bioavailability compared to the reference formulation.

[0013] Another object of the present invention is to provide a xanthophyll composition comprising at least 80% of total xanthophylls, which is further composed of at least 80% by weight of trans-R,R lutein and at least 15% by weight of trans-R,R zeaxanthin.

[0014] Another object of the present invention is to provide a xanthophyll-rich composition composed of trans-R,R zeaxanthin and selective isomers such as trans-R,R lutein, and substantially free of R,S and S,S zeaxanthin.

[0015] Yet another object of the present invention is to provide a composition obtained from extracts of two different plant sources containing xanthophyll esters prepared by treatment with food grade solvent in specific ratios by using specific reaction conditions involving unit operations.

[0016] Another important object of the present invention is to provide a xanthophyll-rich composition prepared from extracts of two different plant sources such as marigold and paprika.

[0017] Another object of the present invention is to provide a process for preparing a xanthophyll composition which is prepared by mixing two xanthophyll ester rich extracts in a certain ratio, treating the reaction mixture with alcohol alkali, followed by neutralization isolation and purification process to obtain a xanthophyll composition of desired purity.

[0018] Another object of the present invention is to provide for selecting particle size by micronization and further adding suitable oil as a vehicle along with optional bioenhancer, solubility enhancer, antioxidant and optional flavoring agent and mixing to obtain a xanthophyll composition in the form of an oil suspension.

[0019] Another object of the present invention is to provide a xanthophyll composition optionally comprising trans-R,R zeaxanthin and trans-R,R lutein and at least one pharma- ceutically and / or nutraceutical acceptable excipient for preparing beadlets.

[0020] Another object of the present invention is to provide a xanthophyll composition that is substantially free of R,S and S,S zeaxanthin and suitable for human consumption for use in various eye and brain health applications. Summary of the Invention [Problem to be solved by the invention]

[0021] According to one aspect of the present invention, i) trans-R,R lutein in the range of 65-85% by weight; ii) trans-R,R zeaxanthin in the range of 10-30% by weight; iii) one or more carriers and / or vehicles in the range of 10-80% by weight; iv) one or more solubility enhancers in the range of 1-10% by weight; v) one or more flavoring agents in the range of 1 to 5% by weight; vi) one or more bioavailability enhancers in the range of 1 to 10% by weight; and vii) one or more antioxidants in the range of 1 to 10% by weight; Trans-R,R lutein and trans-R,R zeaxanthin are extracted from marigold and paprika oleoresins and are present in a ratio of 4:1 to 6:1, preferably 5:1, to provide a xanthophyll composition that is substantially free of R,S and S,S zeaxanthin.

[0022] The invention further relates to processes for preparing the formulations as oil suspensions and beadlets. [Brief description of the drawings]

[0023] According to Figures 1-8, the graph designated as Product A (Micro OS R+) is read as trans-R,R lutein and trans-R,R zeaxanthin composition. The reference is Product B. [Figure 1] Figure 1 shows a comparative linear plot of baseline-corrected mean ± SD of serum trans-R,R lutein concentrations vs. time for Micro OS R+ and reference. Figure 1 reveals that baseline-corrected serum trans-R,R lutein concentrations in the Micro OS R+ group were significantly higher compared to the reference group at 2 to 72 hours post-dose (P<0.05). [Diagram 2] Pharmacokinetic parameters of trans-R,R lutein - Cmax, AUC0-72 and AUC0-T: (Micro OS R+ vs. reference). Figures 2 and 3 reveal that the Micro OS R+ group showed significantly higher absorption of trans-R,R lutein compared to the reference group. This is evident from the 2.5-fold higher Cmax, 2.9-fold higher AUC0-72 and 3.2-fold higher AUC0-T levels. [Diagram 3]Pharmacokinetic parameters of trans-R,R lutein - Cmax, AUC0-72 and AUC0-T: (Micro OS R+ vs. reference). Figures 2 and 3 reveal that the Micro OS R+ group showed significantly higher absorption of trans-R,R lutein compared to the reference group. This is evident from the 2.5-fold higher Cmax, 2.9-fold higher AUC0-72 and 3.2-fold higher AUC0-T levels. [Figure 4] Figure 4 shows a comparative linear plot of baseline-corrected mean ± SD of serum trans-R,R zeaxanthin concentrations vs. time for Micro OS R+ and reference. Figure 4 reveals that baseline-corrected serum trans-R,R zeaxanthin concentrations in the Micro OS R+ group were significantly higher compared to the reference group from 4 to 72 hours post-dose (P<0.05). [Diagram 5] Pharmacokinetic parameters of trans-R,R zeaxanthin - Cmax, AUC0-72 and AUC0-T: (Micro OS R+ vs. reference). Figures 5 and 6 show that the Micro OS R+ group showed significantly higher absorption of trans-R,R zeaxanthin compared to the reference group. This is evident from the 1.7-fold higher Cmax and 2.2-fold higher AUC0-72 and AUC0-T levels. [Figure 6] Pharmacokinetic parameters of trans-R,R zeaxanthin - Cmax, AUC0-72 and AUC0-T: (Micro OS R+ vs. reference). Figures 5 and 6 show that the Micro OS R+ group showed significantly higher absorption of trans-R,R zeaxanthin compared to the reference group. This is evident from the 1.7-fold higher Cmax and 2.2-fold higher AUC0-72 and AUC0-T levels. [Figure 7] FIG. 1 shows the particle size of trans-R,R lutein and trans-R,R zeaxanthin before preparation of oil suspensions. [Figure 8] FIG. 1 shows the particle size of trans-R,R lutein and trans-R,R zeaxanthin after preparation of an oil suspension. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present invention relates to a xanthophyll composition containing selective isomers such as trans-R,R lutein and trans-R,R zeaxanthin, and a process for preparing them. The present invention specifically relates to a xanthophyll composition that accounts for more than 80% of the total xanthophyll. It further contains at least 80% by weight of trans-R,R lutein and at least 15% by weight of trans-R,R zeaxanthin. The process consists of mixing extracts containing xanthophyll esters obtained from two different plant sources in a certain ratio and further processing to obtain a xanthophyll composition selectively containing trans-R,R lutein and trans-R,R zeaxanthin in a desired percentage. The enhanced bioavailability of the xanthophyll composition comprising trans-R,R lutein and trans-R,R zeaxanthin is due to the combined effect of a micronization process to obtain a selective range of particle sizes in a preferred oil as a vehicle, and selective bioenhancers including flavoring agents, solubility enhancers and antioxidants, which surprisingly exerts a 1.5-4 fold improvement in bioavailability over the baseline. This xanthophyll composition comprising the trans-R,R isomer of lutein and the trans-R,R isomer of zeaxanthin in a specific ratio of 4:1-6:1 is further formulated in various forms such as beadlets, powders, oil suspensions, granules, capsules, tablets, films, or any other suitable oral, parenteral or topical dosage forms using pharma- ceutically or nutritionally acceptable excipients and / or carriers. The xanthophyll composition is substantially free of R,S and S,S zeaxanthin and is suitable for human consumption for use in various eye and brain health applications.

[0025] According to an embodiment of the present invention, a xanthophyll composition selectively containing trans-R,R lutein and trans-R,R zeaxanthin in specific weight percentages is prepared from extracts of two different plant sources using food grade solvents and industrially viable processes, and is treated with alkali, neutralized, isolated, purified, and formulated with pharma- ceutical and / or nutraceutical acceptable excipients to enhance bioavailability.

[0026] In the context of the present invention, the term "xanthophyll composition" refers to a composition composed 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, such that the composition is selectively composed of trans-R,R lutein and trans-R,R zeaxanthin. More importantly, the composition is substantially free of the R,S and S,S isomers of zeaxanthin.

[0027] According to a main embodiment of the present invention, the xanthophyll composition selectively comprises at least 80% by weight of total xanthophylls, of which at least 65-85% by weight is trans-R,R lutein and at least 10-30% by weight is trans-R,R zeaxanthin, which is useful for nutrition and health care.

[0028] According to an important embodiment of the present invention, the plant material used for 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 vegetable mallow), as well as various types of pumpkin, paprika, other dark green leafy vegetables, parsley, kale, egg yolk, corn, etc. According to one embodiment of the present invention, the plant material selected for the preparation of the present composition is a hybrid species of marigold flowers (Tagetes japonicus) and paprika. In particular, the lutein ester-rich extract is obtained from marigold flowers and the zeaxanthin ester-rich extract is obtained from certain types of paprika.

[0029] According to another embodiment of the present invention, marigold flowers containing a specific percentage of xanthophyll content are selected and processed (extracted using food grade solvents under specific conditions) to obtain pellets. An extract containing about 14-17% by weight of lutein esters is obtained for further processing.

[0030] According to another embodiment of the present invention, fresh paprika fruit pods having an R,R zeaxanthin content of about 0.04-0.08% by weight are selected and dried to a specific moisture level and trans-R,R zeaxanthin content. The dried peppers (paprika) are cut into small pieces to obtain a powder, which is then treated with a food grade solvent to obtain an extract having 3-4% by weight of R,R zeaxanthin esters.

[0031] Marigold Flower (scientific name - Tagetes persica, Asteraceae): It is a species of the genus Tagetes native to Mexico. It is cultivated in Asia, Africa, and South America. Marigold is also cultivated in Karnataka, especially in the areas of Hassan, Chikmagalur, Mysore, Chamarajanagar, and Davangere. For further processing, the marigold flowers are delivered to facilities in Hassan (Karnataka) and Kangayam (Tamil Nadu), India. Extraction is carried out by manual intervention to prepare a composition that is rich in xanthophylls, which selectively contains trans-R,R zeaxanthin isomers and trans-R,R lutein in specific weight percentages, distinct from the naturally occurring composition of marigold flowers, abundant, and safe for human consumption.

[0032] Another source for use in the present invention is paprika (Capsicum annuum), a species of the plant genus Capsicum (pepper) native to southern North America and northern South America. This species is the most common of the five indigenous peppers and is widely cultivated. For the purposes of the present invention, the raw material may be purchased from outside India or may be cultivated as a pepper in Karnataka.

[0033] The xanthophyll composition comprises at least 80% by weight of total xanthophylls, of which at least 65% by weight is trans-R,R lutein and at least 10% by weight is trans-R,R zeaxanthin.

[0034] According to a preferred embodiment, the composition is substantially free of R,S and S,S zeaxanthin.

[0035] In one embodiment, the xanthophyll composition is comprised of trans-R,R lutein and trans-R,R zeaxanthin in a ratio of 4:1 to 6:1.

[0036] In one embodiment, the xanthophyll composition is composed of trans-R,R lutein and trans-R,R zeaxanthin in a 5:1 ratio.

[0037] In one embodiment, the xanthophyll composition comprises 10-15 mg of trans-R,R lutein and 2-3 mg of trans-R,R zeaxanthin.

[0038] In one embodiment, the xanthophyll composition including trans-R,R lutein and trans-R,R zeaxanthin is in the range of 0.1 to 10 microns.

[0039] In one embodiment, the oil suspension composition obtained according to Example 1 has particle size in the range of 0.1 to 10 microns.

[0040] In one embodiment, the xanthophyll composition comprises a 1.5 to 4 fold increase in bioavailability relative to the reference (Product B).

[0041] According to yet another embodiment of the present invention, the xanthophyll composition described herein can be formulated with at least one pharmaceutical, nutraceutical or food grade excipient or combination thereof.The excipient is selected from bioavailability enhancer, carrier / vehicle, granulating agent, inert core, coating agent, solvent, diluent, binder, lubricant, disintegrant, antioxidant, oil, surfactant, solubilizer, emulsifier, or any other excipient that is known to those skilled in the art as the excipient required to prepare a palatable dosage form that is acceptable to subjects.

[0042] According to a further embodiment of the invention, the bioavailability enhancer is selected from the group consisting of nigella (black cumin oil), caraway (carum) oil, phosphatidylcholine (lecithin), sodium starch octenyl succinate, allspice berry, amber essence, anise seed, arnica, balsam of Peru, basil, bay leaf, benzoin gum, bergamot, bois de rose (rosewood), cajeput, calendula (pot marigold), white camphor, caraway seed, cardamom, carrot seed, cedarwood, celery, chamomile (German or Hungarian), chamomile (Roman or English), cinnamon, citronella, clary sage, clove bud, coriander, cumin, cypress, eucalyptus, fennel, Siberian fir needle, frankincense (olibanum oil), garlic, rose geranium, ginger, grapefruit, hyssop, jasmine absolute. , jojoba, juniper berry, lavender, lemon, lemongrass, lime, sweet marjoram, mugwort, mullein flower, milghum, bigarade neroli, nutmeg, bitter orange, sweet orange, oregano, patchouli, pennyroyal, black pepper, peppermint, petitgrain, pine needle, pokeweed, rose absolute, rosehip seed, rosemary, dalmation sage, sandalwood oil, sassafras, spearmint, spikenard, spruce (hemlock), tangerine, tea tree, thuja (cedar leaf), thyme, vanilla essence, vetiver, wintergreen, witch hazel (hamemeria) extract, or ylang (cananga) extract or combinations thereof.

[0043] According to a preferred embodiment of the present invention, the bioavailability enhancer is selected from Nigella (Black Cumin Oil), Caraway (Caraway) Oil, Phosphatidylcholine (Lecithin).

[0044] According to a further embodiment, the bioavailability enhancer is present in the range of 1-10% by weight of the composition.

[0045] According to a further embodiment, the bioavailability enhancer is present in the range of 1-5% by weight of the composition.

[0046] According to a further embodiment, the bioavailability enhancer is present in the range of 1-3% by weight of the composition.

[0047] According to a preferred embodiment of the present invention, the solubility enhancer may be selected from thyme oil, olive oil, linseed oil (flaxseed oil) or mixtures thereof.

[0048] According to a further embodiment, the solubility enhancer is present in the range of 1-10% by weight of the composition. According to a further embodiment, the solubility enhancer is present in the range of 1-5% by weight of the composition.

[0049] According to further embodiments of the present invention, the antioxidant may be selected from, but is not limited to, butylated hydroxytoluene, butylated hydroxyanisole, ascorbic acid, ascorbyl palmitate, sodium ascorbate, mixed tocopherols, alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol, synthetic racemic tocopherol and tocopherol esters or combinations thereof.

[0050] According to a further embodiment, the antioxidant is present in the range of 1-10% by weight of the composition. According to a further embodiment, the antioxidant is present in the range of 1-10% by weight of the composition. According to a further embodiment, the antioxidant is present in the range of 1-5% by weight of the composition. According to a further embodiment, the antioxidant is present in the range of 1-3% by weight of the composition.

[0051] According to further embodiments of the present invention, the carrier / vehicle may be selected from, but is not limited to, sunflower oil, coconut oil, corn oil, 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 combinations thereof.

[0052] According to further embodiments of the present invention, the hydrophilic carrier may be selected from, but is not limited to, celluloses such as alkylcelluloses (methylcellulose), hydroxyalkylcelluloses (e.g., hydroxymethylcellulose, hydroxypropylcellulose), carboxyalkylcelluloses (e.g., carboxymethylcellulose and its alkali metal salts, e.g., sodium salt), carboxyalkylalkylcelluloses (e.g., carboxymethylethylcellulose), carboxyalkylcellulose esters (e.g., carboxymethylcellulose butyrate, carboxymethylcellulose propionate, carboxymethylcellulose acetate butyrate and carboxymethylcellulose acetate propionate) or combinations thereof; Polyacrylates, such as polymethacrylates, methacrylate copolymers (e.g., methacrylic acid-methyl methacrylate copolymer, dimethylaminoethyl methacrylate-butyl methacrylate-methyl methacrylate copolymer, and diethylaminoethyl methacrylic acid-methyl methacrylate copolymer), and methacrylic acid copolymers (e.g., methacrylic acid-ethacrylic acid copolymer), or combinations thereof; povidones, such as polyvinylpyrrolidone, polyvinyl acetate esters (e.g., polyvinyl acetate phthalate (PVAP), and polyethylene glycol polyvinyl acetate copolymers (e.g., polyethylene glycol-polyvinyl caprolactam-polyvinyl acetate copolymer), or combinations thereof; Starch and starch derivatives, such as modified starch, corn starch, potato starch, pregelatinized starch, dextrin, acid treated starch, alkali treated starch, bleached starch, oxidized starch, enzyme treated, monostarch phosphate, distarch phosphate, phosphate distarch phosphate, acetylated distarch phosphate, starch acetate, acetylated distarch adipate, hydroxypropyl starch, hydroxypropyl distarch phosphate, hydroxypropyl distarch glycerol, sodium starch octenyl succinate, acetylated oxidized starch and maltodextrin or combinations thereof; Gums such as alginate pectin, carrageenan agar, gum arabic, tragacanth gum, karaya gum, ghatti gum, guar gum, locust bean gum, tara gum, xanthan gum, gellan gum and welan gum or combinations thereof; sugars such as fructans, sucrose, glucose and fructose or combinations thereof.

[0053] According to a preferred embodiment, the carrier is MCT oil. According to a further embodiment, the MCT oil is present in the range of 20-80% by weight of the composition. According to a further embodiment, the MCT oil is present in the range of 50-70% by weight of the composition.

[0054] According to a preferred embodiment, the carrier is a modified starch. According to a further embodiment, the starch is present in the range of 10-40% by weight of the composition. According to a further embodiment, the starch is present in the range of 10-30% by weight of the composition. According to a further embodiment, the starch is present in the range of 10-20% by weight of the composition.

[0055] According to one embodiment, the xanthophyll composition containing trans-enriched lutein and trans-R,R isomers of zeaxanthin in a specific ratio can be further formulated in different forms such as beadlets, powders, oil suspensions, granules, capsules, tablets, films or any other suitable oral, parenteral or topical dosage forms, preferably oil suspensions and beadlets, using pharma- ceutical or nutritionally acceptable excipients and / or carriers.

[0056] According to one embodiment of the present invention, the process for producing beadlets involves spraying a carotenoid emulsion into a square chamber and embedding it in a cold matrix bed, which provides continuous rotation and circulation of cold water through a jacketed vessel.

[0057] According to one embodiment of the present invention, the process for preparing a carotenoid emulsion comprises a solvent phase and an aqueous phase, which are mixed under stirring under specific conditions to obtain an emulsion. The solvent phase described herein is prepared by dissolving an active substance in a solvent phase comprising an organic solvent, an antioxidant, and at least one or more oily excipients under specific conditions, and filtering it to remove any insoluble residues. The aqueous phase described herein is prepared by dissolving a hydrophilic carrier and an antioxidant in water.

[0058] According to another embodiment of the present invention, the solvent phase is added to the water phase under stirring to obtain an emulsion, which is then homogenized in a high pressure homogenizer.The homogenized emulsion is then evaporated under vacuum to obtain a solvent-free carotenoid emulsion.

[0059] According to one important embodiment of the present invention, the process for producing carotenoid beadlets involves spraying a carotenoid emulsion into a square chamber, which is embedded in a bed of cold matrix and allowed to solidify to form beadlets.

[0060] According to another embodiment of the invention, the process for producing carotenoid beadlets is carried out in a square chamber with two inlets at the top for the emulsion spray nozzle and one for the hydrophilic matrix, which also has two outlets at the bottom of the chamber for collecting the beadlets and unused matrix bed.

[0061] According to another important embodiment of the present invention, the square chamber provides continuous rotation as well as circulation of cold water through a jacketed vessel to maintain the internal temperature below 20°C.

[0062] According to the main embodiment of the present invention, the process for producing carotenoid beadlets involves loading a matrix bed through one inlet at the top of a square chamber and keeping it cool with the help of cold water circulation. The emulsion is sprayed from the spray nozzles in the rotating square chamber and the droplets embed into the particles of the cold matrix bed to form beadlets.

[0063] The beadlets formed by the above process are collected from an outlet at the bottom of the square chamber and subjected to drying until the desired moisture level is achieved.

[0064] According to another embodiment of the invention, the matrix bed loaded into the square spray chamber is selected from, but not limited to, starch derivatives such as waxy maize starch, modified starches in combination with starch and talc, silicic acid, hydrogenated fats, metal salts of higher fatty acids such as calcium stearate, alkali metal / alkaline earth metal aluminum silicates, sodium or calcium aluminum silicate, alumina, calcium silicate, powdered silica gel, magnesium carbonate and magnesium oxide or combinations thereof.

[0065] According to a preferred embodiment of the present invention, the process for preparing a xanthophyll composition in oil suspension form includes obtaining trans-R,R lutein and trans-R,R zeaxanthin in a selective ratio of 4:1 to 6:1 by extraction from marigold and paprika oleoresin. The marigold and paprika are extracted to obtain trans-R,R lutein and trans-R,R zeaxanthin (called concentrate) which are further micronized by using an air jet mill to obtain a particle size in the range of 0.1 to 10 microns. The obtained selective ratio of micronized trans-R,R lutein and trans-R,R zeaxanthin having a preferred particle size is mixed by adding bioavailability enhancers, solubility enhancers, antioxidants, carrier oils, and optionally flavorings using a ball mill at room temperature. The obtained oil suspension composition has a particle size in the range of 0.1 to 10 microns.

[0066] The particle size of trans-R,R lutein and trans-R,R zeaxanthin ranges from 0.1 to 10 microns.

[0067] Other excipients such as carrier oils / vehicles, bioavailability enhancers, solubility enhancers, stabilizers, and antioxidants are added to the ball mill.

[0068] The resulting oily suspension composition according to Example 1 has a particle size in the range of 0.1 to 10 microns.

[0069] According to a further embodiment, a xanthophyll composition in the form of an oil suspension comprising xanthophyll and a pharma- cetically and / or nutritionally acceptable excipient provides reduced bioavailability enhancement.

[0070] At the outset of the following description, it should be understood that the following description is merely illustrative of certain forms of the invention, however such specific forms are merely exemplary embodiments and are not intended to be construed as limiting in any way to imply any limitations on the scope of the invention.

[0071] Example 01: According to a preferred embodiment, different size batches from small to large were run to support the industrial feasibility of the manufacturing process. Preparation of oil suspension composition; batch size 500 g [Table 1]

[0072] Weight of marigold oleoresin (33.5 kg) containing 16.6% xanthophyll content (by spectrophotometry) and 13.6 lutein content (by HPLC method). Weight of paprika oleoresin (16.5 kg) containing 5.7% xanthophyll content (by spectrophotometry) and 3.5% zeaxanthin content (by HPLC method).

[0073] Each oleoresin was added to a reactor containing an alcoholic alkaline aqueous solution (15 litres of water and 62.5 litres of ethanol containing 10 kg of potassium hydroxide). The mixture was heated in the reactor at 80°C for 4 hours with stirring. During the reaction, the extent of hydrolysis was monitored by HPLC and the reaction was stopped by adding 250 litres of water. The reaction mass was homogenized at room temperature and ethyl acetate (300ltr) was added to the reactor for liquid-liquid extraction and this process was repeated 3-5 times. The EA layer was separated and washed with water (1:1) at room temperature. The ethyl acetate washed with water was further taken for concentration to obtain concentrated material. The concentrated material was isolated using hexane to remove impurities and obtain semi-purified crystals of xanthophyll composition (22.5 kg). The obtained semi-purified crystals were purified with 5 volumes of ethanol at room temperature and then filtered (7.8 kg). The obtained crystals were dried in vacuum at a temperature of about 50-55°C for 36 hours. The yield of dried xanthophyll crystals was 9.6 kg (9.6%). The xanthophyll content was 87.2% by weight (UV / Vis-spectrophotometric), of which the contents of trans-R,R lutein and trans-R,R zeaxanthin were 70.3% and 14.4%, and the area percentages of trans-R,R lutein and trans-R,R zeaxanthin were 80.6% and 16.5%, respectively, as determined by normal phase HPLC analysis.

[0074] The resulting concentrate (2.5 kg) containing 87.2% xanthophyll content (by spectrophotometry) was taken for micronization. The micronization of marigold paprika extract concentrate having a particle size (DV90) of 1690 μm was micronized using an air jet mill. The resulting micronized crystals (2.2 kg) had a particle size of (DV50) 1.61μm and (DV90) 3.24 μm (Figure 8) and the xanthophyll content was 89.09% by weight (UV / Vis-spectrophotometric), of which the contents of trans-R,R lutein and trans-R,R zeaxanthin were 71.62% and 14.75%, and the 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.

[0075] A formulation was made as an oil suspension using micronized marigold paprika extract concentrate (170 g) containing 87.2% (spectrophotometric and HPLC) xanthophyll content. The micronized marigold paprika extract concentrate was mixed with 14% (70 g) of other ingredients such as olive oil, thyme oil, linseed oil (flaxseed oil), D-limonene, lecithin (phosphatidylcholine) (obtained from sunflower) by adding medium chain triglyceride (MCT) oil (260 g) and tocopherols by using a ball mill at room temperature. The mixture was sieved and unloaded (500 g) to determine the particle size of the contained. (DV50) 1.04μm and (DV90) 2.66 μm (Figure 7) and the xanthophyll content was 28.05 wt% (UV / Vis-spectrophotometric), of which the contents of trans-R,R lutein and trans-R,R zeaxanthin were 22.64% and 4.63%, and the 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.

[0076] Example 02: Preparation of oil suspension; batch size 18Kg [Table 2]

[0077] A weighted amount of micronized marigold paprika extract concentrate (6.28 kg) containing 80.97% xanthophyll content (by spectrophotometric and HPLC methods) was used to make the formulation as an oil suspension. The micronized marigold paprika extract concentrate was mixed with 14% (2.52 kg) of other ingredients such as olive oil, thyme oil, linseed oil (flaxseed oil), D-limonene, lecithin (phosphatidylcholine) (obtained from sunflower) by adding medium chain triglyceride (MCT) oil (9.2 kg) and tocopherols by using a ball mill at room temperature. The mixture was sieved, unloaded (18 kg) and analyzed for particle size content (DV90) of 2.62 μm and the xanthophyll content was 28.23 wt% (UV / Vis-spectrophotometric), of which the contents of trans-R,R lutein and trans-R,R zeaxanthin were 22.6% and 4.71%, and the 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.

[0078] Example 03: Preparation of beadlets (15%); batch size 24.2 kg [Table 3]

[0079] procedure: A. Micronized marigold paprika extract concentrate was dissolved in methylene dichloride, followed by the addition of tocopherol with stirring. The solution was heated at 40-45°C for 30 minutes until the solution became clear, and then cooled to room temperature. The solution was filtered to obtain a clear brownish colored solution. B. The modified starch is dissolved in water and heated to 50-55°C with stirring until a clear solution is obtained. The aqueous system is cooled to room temperature. C. The solution from stage A was mixed into the solution from stage B with stirring and sodium ascorbate was added to the mixture. D. The solution from stage C was homogenized at 15,000–20,000 rpm for 5 min intervals. E. The solution from Stage D was added to a rotary evaporator and the methylene dichloride was removed by rotating at 4 rpm under vacuum at 60° C. in a water bath and cooled to room temperature. F. 3.5 kg starch, first dried at 105° C. for 4 hours, was added to the spray pan and dry ice was added to the chamber to reduce the temperature to 2° C. Spraying of the solution from Stage E was started onto the fluidized dry starch. G. Coated macular carotenoid beadlets were collected by sieving the material from stage F, drying the beadlets in a tray oven at 35 °C-40 °C, and sieving the dried beadlets through mesh 60 and 40 sieves.

[0080] Example 04: Preparation of beadlets (20%); batch size 31.7 kg [Table 4]

[0081] procedure: A. Micronized marigold paprika extract concentrate was dissolved in dichloromethane and then tocopherol was added with stirring. The solution was heated at 40-45°C for 30 minutes until it became clear, and then cooled to room temperature. The solution was filtered to obtain a clear brownish solution. B. The modified starch was dissolved in water and heated to 50-55°C with stirring until a clear solution was obtained. The aqueous system was cooled to room temperature. C. The solution from stage A was mixed into the solution from stage B with stirring and sodium ascorbate was added to the mixture. D. The solution from stage C was homogenized at 15,000–20,000 rpm for 5 min intervals. E. The solution from Stage D was added to a rotary evaporator and the methylene dichloride was removed by rotating at 4 rpm under vacuum at 60° C. in a water bath and cooled to room temperature. F. 3.5 kg starch, first dried at 105° C. for 4 hours, was added to the spray pan and dry ice was added to the chamber to reduce the temperature to 2° C. Spraying of the solution from Stage E was started onto the fluidized dry starch. G. Coated macular carotenoid beadlets were collected by sieving the material from stage F, drying the beadlets in a tray oven at 35 °C-40 °C, and sieving the dried beadlets through mesh 60 and 40 sieves.

[0082] Example 05: Preparation of beadlets (10%); batch size 19.5 kg [Table 5]

[0083] procedure: A. Disperse the modified starch and sodium ascorbate binder in warm water (50-55°C) with stirring to form a clear solution. B. Micronized marigold paprika extract concentrate and tocopherol were added to the dispersion of stage A with stirring and allowed to form a solution for 15 minutes. C. The solution from stage B was processed with a Polytron high shear mixer, mixing the cake uniformly into the binder solution at 5000-12000 RPM for 15 minutes, and observing the temperature and flow ability of the solution. D. The solution from stage C was added to the inlet of a horizontal grinder and processed in repeated passes to reduce size and properly emulsify the solution using the following processing parameters: E. After passing through the ATFE, the solution is entrapped in Starch N200 which is fluidized at the desired process parameters. F. Dry the beadlets in a 55°C oven to achieve an LOD of less than 5%.

[0084] The bioavailability study design of a xanthophyll composition containing trans-R,R lutein and trans-R,R zeaxanthin in the form of an oil suspension was carried out as Study 1 below:

[0085] Test 1: For ease of understanding, the sample designation for Product A is Micro OS R+ and Product B is the reference sample. Furthermore, in the examples where lutein and zeaxanthin are mentioned, it shall be considered as trans-R,R lutein and trans-R,R zeaxanthin.

[0086] the purpose: To compare the bioavailability of 10 mg trans-R,R lutein and 2 mg trans-R,R zeaxanthin from a single oral dose (oil suspension) as Product A (Micro OS R+) with Product B (reference) formulation (10 mg trans-R,R lutein and 2 mg trans-R,R zeaxanthin / capsule) in supplementation in healthy adult human subjects under fed conditions. Product B is the commercial product Florglo® (trans-R,R lutein) and Optisharp™ (trans-R,R zeaxanthin).

[0087] Study design: double-blind, balanced, randomized, two-treatment, single-phase, single-dose, parallel study Preparation: Micro OS R+ sample: Prepared according to Example 01 in a 5:1 ratio. Product B: Commercially available products Florglo and Optisharp (FloraGLO® Lutein 20% SAF and OPTISHARP™ Natural (Zeaxanthin) 14% Safflower Oil) were calculated and mixed to maintain a 5:1 ratio of 10 mg trans-R,R Lutein and 2 mg trans-R,R Zeaxanthin per capsule. Number of volunteers: 90 healthy volunteers (45 volunteers / group) Housing of Volunteers: Volunteers were housed for 7 days before and after the study to control the diet containing minimal amounts of lutein and zeaxanthin.

[0088] Pharmacokinetic (PK) assessment: Baseline-corrected serum concentrations of trans-R,R lutein and trans-R,R zeaxanthin were measured at various time points. Time points (hours): -48 hours, -24 hours, 0 hours, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 24 hours, 48 ​​hours, and 72 hours. -Pharmacokinetic parameters (Ln(Cmax), Ln(AUC0-72), Ln(AUC0-T and Tmax)) -90%CI, T / R ratio

[0089] Bioanalytical Procedures: Lutein and zeaxanthin (lutein including trans-R,R lutein and zeaxanthin including trans-R,R zeaxanthin) were estimated in serum using a validated HPLC method.

[0090] Results for trans-R,R Lutein and trans-R,R Zeaxanthin Baseline-corrected serum trans-R,R lutein concentrations in product A (Micro OS R+) and product B (reference) [Table 6] *P value < 0.05 -Baseline-corrected serum trans-R,R lutein concentrations in the Micro OS R+ group were significantly higher than those in the reference group at all time points (2 to 72 hours) after dosing (P<0.05) (Figure 1). -Baseline-corrected serum trans-R,R lutein concentrations in the Micro OS R+ group showed more than two-fold higher absorption at all time points and three-fold higher absorption at Tmax (20 hours) compared to the reference group.

[0091] Baseline-corrected pharmacokinetic parameters of trans-R,R lutein [Table 7] $Median(min-max) *P value < 0.05 The -Micro OS R+ group demonstrated significantly higher absorption of trans-R,R lutein compared to the reference group (P<0.05), as evidenced by significantly higher baseline-corrected log-transformed Cmax, AUC0-72, and AUC0-T. The -Micro OS R+ group reached the highest serum trans-R,R lutein concentration (Tmax) at 20 hours, while the reference group achieved the highest serum trans-R,R lutein concentration at 16 hours. - The 90% confidence intervals for Cmax and AUC parameters were significantly higher than the bioequivalence criteria of 80-125%, revealing a significantly better bioavailability in the Micro OS R+ group than in the reference group.

[0092] Fold of study compared with reference for baseline-corrected trans-R,R-lutein PK parameters. [Table 8] The -Micro OS R+ group demonstrated significantly higher absorption of trans-R,R lutein compared to the reference group, as evidenced by 2.5-fold higher Cmax, 2.9-fold higher AUC0-72, and 3.2-fold higher AUC0-T levels (Figures 2 and 3).

[0093] Results for trans-R,R zeaxanthin Baseline-corrected serum trans-R,R zeaxanthin concentrations for Product A (Micro OS R+) and Product B (reference) [Table 9] *P value < 0.05 -Baseline-corrected serum trans-R,R zeaxanthin concentrations in the Micro OS R+ group were significantly higher than those in the reference group at 4 to 72 hours after administration (P<0.05) (Figure 4). -Baseline-corrected serum trans-R,R zeaxanthin concentrations in the Micro OS R+ group demonstrated greater than 1.5-fold higher absorption at all time points and 2.9-fold higher absorption at Tmax (20 hours) compared to the reference group.

[0094] Baseline-corrected pharmacokinetic parameters of trans-R,R zeaxanthin [Table 10] $Median(min-max) *P value < 0.05 The -Micro OS R+ group demonstrated significantly higher absorption of trans-R,R zeaxanthin compared to the reference group (P<0.05), as evidenced by significantly higher baseline-corrected log-transformed Cmax, AUC0-72, and AUC0-T. -The Micro OS R+ group reached the highest serum trans-R,R zeaxanthin concentration (Tmax) at 20 hours, while the reference group achieved the highest serum concentration at 24 hours. The 90% confidence intervals for Cmax and all AUC parameters were significantly higher than the bioequivalence criteria of 80–125%, revealing significantly better bioavailability of trans-R,R-zeaxanthin in the Micro OS R+ group than in the reference group.

[0095] Fold of study compared to reference for baseline-corrected trans-R,R-zeaxanthin PK parameters. [Table 11] -Compared to the reference group, the Micro OS R+ group demonstrated significantly higher absorption of trans-R,R zeaxanthin, as evidenced by a 1.7-fold higher Cmax and 2.2-fold higher AUC0-72 and AUC0-T levels.

[0096] Conclusions of the study on the composition of the oil suspension (Micro OS R+; Product A): The Micro OS R+ (Product A) composition demonstrated significantly higher absorption of trans-R,R lutein, ranging from 2.9-3.2-fold, and trans-R,R zeaxanthin, 2.2-fold, at multiple time points measured after administration when compared to the reference (Product B) composition in terms of achieving higher serum concentrations. The higher absorption, 90% confidence intervals for Cmax, and all AUC parameters were significantly higher than the bioequivalence criteria of 80-125%, revealing a significantly better bioavailability of the Micro OS R+ group than the reference group. [Table 12]

Claims

1. 1. A xanthophyll composition comprising: a) 35% by weight of trans-R,R lutein and trans-R,R zeaxanthin; b) 51% by weight of one or more carriers and / or vehicles; c) 8% by weight of one or more solubility enhancers; d) 2% by weight of one or more flavoring agents; e) 2% by weight of one or more bioavailability enhancers; and f) a xanthophyll composition comprising one or more antioxidants in the range of 2% by weight, A xanthophyll composition in which trans-R,R lutein and trans-R,R zeaxanthin are extracted from marigold and paprika oleoresins and are present in a ratio of 4:1 to 6:1, and which is substantially free of R,S zeaxanthin and S,S zeaxanthin.

2. 2. The xanthophyll composition of claim 1, wherein the particle size of the trans-R,R lutein and trans-R,R zeaxanthin ranges from 0.1 to 10 microns.

3. 2. The xanthophyll composition of claim 1, wherein the carrier and / or vehicle is selected from medium chain triglyceride (MCT) oil, sunflower oil, coconut oil, corn oil, cottonseed oil, canola oil, palm oil, peanut oil, safflower oil, sesame oil, soybean oil, rapeseed oil and medium chain triglycerides or combinations thereof, preferably MCT oil.

4. 2. The xanthophyll composition of claim 1, wherein the solubility enhancer is selected from thyme oil, olive oil, linseed oil (flaxseed oil) or a mixture thereof.

5. 2. The xanthophyll composition of claim 1, wherein the bioavailability enhancer is selected from nigella (black cumin oil), caraway (caraway) oil, phosphatidylcholine (lecithin), or a mixture thereof.

6. 2. The xanthophyll composition of claim 1, wherein the antioxidant is selected from tocopherol and sodium ascorbate or a mixture thereof.

7. 2. The xanthophyll composition of claim 1, wherein the particle size of the resulting composition ranges from 0.1 to 10 microns.

8. 2. The xanthophyll composition of claim 1, wherein the composition has a 1.5 to 4 fold increase in bioavailability over the reference.

9. 2. The xanthophyll composition of claim 1, wherein the composition has a 2-4 fold increase in serum concentration over a reference.

10. 2. The xanthophyll composition of claim 1, wherein the formulation of the xanthophyll composition is in the form of beadlets, powders, oil suspensions, granules, capsules, tablets, or films.

11. 10. The xanthophyll composition of claim 1, wherein the composition is in the form of an oily suspension.

12. A method for producing the xanthophyll composition of claim 10, comprising the steps of: The preparation of said oily suspension comprises the following steps: a) obtaining trans-R,R lutein and trans-R,R zeaxanthin in a selective ratio of 4:1 to 6:1 from marigold and paprika oleoresins according to claim 1; b) micronizing the trans-R,R lutein and trans-R,R zeaxanthin of step (a) by using an air jet mill to obtain a particle size in the range of 0.1 to 10 microns; c) the micronized trans-R,R lutein and trans-R,R zeaxanthin obtained in step (b) is mixed with a carrier and / or vehicle; d) adding a solubility enhancer to step (c) along with the flavoring agent; e) adding a bioavailability enhancer and an antioxidant to step (d); f) mixing the suspension obtained in step (e) using a ball mill to obtain a homogenous oily suspension; g) sieving the mixture resulting from step (f) to obtain an oil suspension of the resulting particle size; A method for producing a xanthophyll composition comprising:

13. A method for producing the xanthophyll composition of claim 10, comprising the steps of: The preparation of the beadlets involves the following steps: a) obtaining trans-R,R lutein and trans-R,R zeaxanthin in a selective ratio of 4:1 to 6:1 from marigold and paprika oleoresins according to claim 1; b) micronizing the trans-R,R lutein and trans-R,R zeaxanthin of step (a) by using an air jet mill to obtain a particle size in the range of 0.1 to 10 microns; c) dissolving micronized trans-R,R lutein and trans-R,R zeaxanthin in methylene dichloride, then dissolving in antioxidants and optionally bioavailability enhancers, warming, then cooling and filtering the solution to obtain a clear brownish colored solution; d) dissolving the modified starch in water, warming until a clear solution is obtained, and cooling to room temperature; e) mixing the solution of stage (c) with the solution of stage (d) under stirring and adding sodium ascorbate to the mixture under homogenization; f) adding the solution of step (e) to a rotary evaporator, removing the methylene dichloride under vacuum, and cooling; g) spraying the solution of step (f) onto dry starch under fluidization; h) collecting and drying the coated macular carotenoid beadlets; A method for producing a xanthophyll composition comprising:

14. A method for producing the xanthophyll composition of claim 10, comprising the steps of: The preparation of the beadlets involves the following steps: a) trans-R,R lutein and trans-R,R zeaxanthin are obtained in a selective ratio of 4:1 to 6:1 from marigold and paprika oleoresin according to claim 1; b) micronizing the trans-R,R lutein and trans-R,R zeaxanthin of step (a) by using an air jet mill to obtain a particle size in the range of 0.1 to 10 microns; c) dispersing the modified starch and sodium ascorbate in warm water to form a clear solution; d) adding micronized tocopherol to the dispersion of step (c); e) processing under a Polytron high shear mixer for uniform mixing and passing through a horizontal grinder for size reduction; A method for producing a xanthophyll composition comprising:

15. 2. The xanthophyll composition of claim 1, wherein the ratio of trans-R,R lutein and trans-R,R zeaxanthin is 5:1.

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