Prolycopene-rich composition and method for using the same

Abstract

The present invention provides a composition that possesses antioxidant activity while having high bioavailability. [Solution] The present invention provides a composition containing 1 to 15% by weight of prolycopene, and a method of using the composition, such as to prevent or treat a condition related to oxidative stress.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 021,188, entitled "PRO - LYCOPENE RICH COMPOSITION AND METHODS OF USING SAME", filed on May 7, 2020, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention generally relates to the field of carotenoids and methods of using them, such as for reducing oxidative stress.

Background Art

[0003] It is well established that topical compositions of various protective ingredients can achieve protection of tissues (e.g., skin) from oxidative stress and ultraviolet radiation. Certain groups of protective compositions are intended for oral administration. These oral compositions contain active ingredients that are delivered to tissues (e.g., skin) via internal transport mechanisms, thus protecting the skin from oxidative stress and / or ultraviolet damage. A particular group of active ingredients suitable for use with such oral compositions are carotenoids. The use of carotenoid mixtures, in which canthaxanthin is the primary carotenoid in the composition, has been described. However, the use of canthaxanthin is known to be limited by its potential adverse effects on pigmentation. Furthermore, various food and beverage products intended to provide protection of the skin from the sun's ultraviolet radiation have been reported. These food and beverage products contain carotenoids as well as ascorbic acid, tocopherol, coenzyme Q10, and reduced glutathione. Compositions for protecting the skin from ultraviolet radiation and its harmful effects, containing provitamin A carotenoids and lycopene, have also been described. The use of such compositions is limited by the potential negative effects that provitamin A carotenoids may have on the health of subjects at certain dosage levels. Excess vitamin A produced in the body from provitamin A carotenoids has been found to have harmful effects on health. In particular, the protective effect of tomato paste, known to contain lycopene, β-carotene, and tocopherol, against UV-induced erythema has been observed. However, problems in achieving desired carotenoid serum levels have been reported, suggesting that it has poor bioavailability.

[0004] Therefore, there has been a long-standing need to develop compositions that possess both high bioavailability and antioxidant activity (for example, the ability to reduce oxidative stress). [Overview of the project]

[0005] According to a first embodiment, a composition is provided comprising a prolycopene in an amount of 1 to 15% by weight and an acceptable carrier.

[0006] In another embodiment, a method is provided for preventing or treating an oxidative stress-related condition or disease in a subject requiring such treatment, comprising administering a therapeutically effective amount of the composition of the present invention to the subject, thereby preventing or treating an oxidative stress-related condition or disease in the subject.

[0007] In some embodiments, the composition further comprises additional carotenoids selected from the group consisting of neuropolene, phytoene, phytofluene, ζ-carotene, β-carotene, trans-lycopene, and any combination thereof.

[0008] In some embodiments, prolycopene constitutes at least 40% (w / w) of the total lycopene in the composition.

[0009] In some embodiments, the weight / weight ratio of prolycopene to translycopene is in the range of 1.5:1 (w / w) to 6:1 (w / w).

[0010] In some embodiments, the composition contains neuropolene in an amount of 3 to 8% by weight of the composition.

[0011] In some embodiments, the composition contains phytoene in an amount of 40 to 50% by weight of the composition.

[0012] In some embodiments, the composition contains phytofluene in an amount of 10 to 20% by weight of the composition.

[0013] In some embodiments, the composition contains 15 to 25% by weight of ζ-carotene.

[0014] In some embodiments, the weight / weight ratio of both phytoene:phytofluene and ζ-carotene is in the range of 2:1 (w / w) to 4:1 (w / w).

[0015] In some embodiments, the composition comprises less than 5% by weight of trans-lycopene, less than 3% by weight of β-carotene, or a combination thereof.

[0016] In some embodiments, the composition further comprises tocopherol.

[0017] In some embodiments, the composition contains tocopherol in an amount of 2 to 5.5% by weight of the composition.

[0018] In some embodiments, the composition contains prolycopene in an amount of 10-20% (w / w) of the total carotenoids in the composition.

[0019] In some embodiments, the composition is intended for use in reducing oxidative stress.

[0020] In some embodiments, administration includes oral administration.

[0021] Unless otherwise specified, all technical and / or scientific terms used herein have the same meaning as generally understood by those skilled in the art to which the present invention pertains. Methods and materials similar to or equivalent to those described herein may be used in carrying out or testing embodiments of the present invention, but exemplary methods and / or materials are described below. In case of any conflict, the present specification, including definitions, shall prevail. Furthermore, materials, methods and examples are merely illustrative and not necessarily limiting.

[0022] Further embodiments and the full scope of the present invention will become apparent from the detailed description given below. However, it should be understood that the detailed description and specific examples illustrating preferred embodiments of the present invention are given merely as examples, as various variations and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

[0023] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by referring to the figures and considering the following detailed description.

Brief Description of the Drawings

[0024] [Figure 1] Includes a graph showing the dose response of tangelo extract (TG) and a control (e.g., Lyc-O-Mato) in the induction of the antioxidant response element (ARE) in normal human dermal fibroblasts (NHDF). [Figure 2] Figures 2A - 2B include bar graphs showing the dose response of TG extract (2A) and Lyc-O-Mato (2B) in the induction of ARE in T47D breast cancer cells. EA: ethyl acetate extraction, SCE: supercritical extraction. [Figure 3] Includes a chromatogram of the HPLC analysis of the TG supercritical extract. A C30 chromatography column was used. [Figure 4] Includes a graph showing the various carotenoid spectra in tangelo that can protect the skin from UV / visible light irradiation with a wide wavelength (WL) in the range of 240 - 520 nm. Phytoene (290 nm), phytofluene (350 nm), ζ-carotene (400 nm), neurosporene (440 - 470 nm), and lycopene (450 - 520 nm).

Modes for Carrying Out the Invention

[0025] In some embodiments, the present invention relates to a composition comprising a rich amount of prolycopene.

[0026] As used herein, the term "prolycopene" refers to tetra-cis-lycopene, (7Z,9Z,7’Z,9’Z)-xi,xi-carotene, 7,9,7’,9’-tetracis-lycopene, or any combination thereof.

[0027] In some embodiments, the supercritical extract of tangerine tomatoes can be standardized by the total carotenoid content. In some embodiments, the standardized extract contains 10–30% by weight of total carotenoids. [Table 1]

[0028] In some embodiments, the compositions of the present invention contain prolycopene in amounts of 3 to 10% by weight, 1 to 15% by weight, 3 to 8% by weight, 4 to 9% by weight, 1 to 6% by weight, 2 to 10% by weight, or 7 to 12% by weight. Each possibility represents a distinct embodiment of the present invention.

[0029] In some embodiments, the composition further comprises additional carotenoids. As used herein, “additional carotenoids” refers to any carotenoid other than or different from prolycopene or its metabolites. In some embodiments, the additional carotenoids are selected from neurosporene, phytoene, phytofluene, ζ-carotene, β-carotene, trans-lycopene, or any combination thereof.

[0030] In some embodiments, the additional carotenoid is neuropolene.

[0031] In some embodiments, prolycopene constitutes at least 40% (w / w) of the total lycopene in the composition, at least 50% (w / w) of the total lycopene in the composition, at least 60% (w / w) of the total lycopene in the composition, at least 70% (w / w) of the total lycopene in the composition, at least 80% (w / w) of the total lycopene in the composition, or at least 90% (w / w) of the total lycopene in the composition, or any value or range in between. Each possibility represents a separate embodiment of the present invention. In some embodiments, prolycopene constitutes 40-90% (w / w) of the total lycopene in the composition, 40-80% (w / w) of the total lycopene in the composition, 50-75% (w / w) of the total lycopene in the composition, 35-75% (w / w) of the total lycopene in the composition, 60-95% (w / w) of the total lycopene in the composition, 55-80% (w / w) of the total lycopene in the composition, or 70-95% (w / w) of the total lycopene in the composition. Each possibility represents a separate embodiment of the present invention.

[0032] As used herein, the term "total lycopene" refers to the amount, weight, quantity, concentration, or level of all pro-(e.g., cis-) and trans-lycopene isomers combined into one.

[0033] In some embodiments, the trans-lycopene is all-trans-lycopene.

[0034] In some embodiments, the weight / weight ratio of prolycopene:trans-lycopene is in the range of 1.5:1(w / w) to 6:1(w / w), 1.5:1(w / w) to 5:1(w / w), 1.5:1(w / w) to 4:1(w / w), 1.5:1(w / w) to 3:1(w / w), or 1.5:1(w / w) to 2:1(w / w). Each possibility represents a distinct embodiment of the present invention.

[0035] In some embodiments, carotenoids are natural carotenoids extracted, isolated, or purified from fruits, vegetables, or plants (including parts of any plant). In another embodiment, carotenoids are carotenoids extracted from tomato plants. In another embodiment, carotenoids are carotenoids extracted from tomato fruits. In another embodiment, tomato carotenoids are tomato extracts concentrated for carotenoids. In another embodiment, tomato carotenoids are tomato extracts high in whole natural carotenoids. In another embodiment, tomato carotenoids are tomato carotenoid complexes. In another embodiment, tomato carotenoid complexes include a complex of plant nutrients containing multiple carotenoids (such as phytoenes, phytofluenes, ζ-carotene, and β-carotene), tocopherols, and phytosterols. In some embodiments, carotenoids are synthetic carotenoids.

[0036] In some embodiments, the present invention provides a tomato extract obtained by an innovative extraction protocol. This particular extract, containing prolycopene (in the amount specified herein), has high bioavailability and antioxidant stress activity. In some embodiments, the high bioavailability and antioxidant stress activity are compared to other tomato extracts. In some embodiments, the high bioavailability and antioxidant stress activity can be provided to subjects requiring the composition of the present invention at lower doses without reducing, for example, the subject's survival, health, or both, while achieving a superior antioxidant stress effect. In some embodiments, administration to subjects requiring the composition of the present invention can enhance the efficacy of the treatment by providing active ingredients such as prolycopene without reducing the subject's survival, health, or both, while increasing the therapeutic effect (antioxidant stress effect) (resulting from the high bioavailability of prolycopene).

[0037] In some embodiments, the compositions of the present invention provide a larger amount of carotenoids, such as prolycopene, which have greater bioavailability compared to extracts derived from other plants, fruits, or vegetables, such as tomatoes. In some embodiments, the compositions of the present invention provide high therapeutic efficacy along with high bioavailability compared to extracts derived from other plants, fruits, or vegetables, such as tomatoes.

[0038] In some embodiments, the compositions of the present invention include natural carotenoids, synthetic carotenoids, or any combination thereof.

[0039] In some embodiments, the composition contains phytoene in amounts of 40-60% by weight, 35-55% by weight, 40-55% by weight, 45-55% by weight, 40-50% by weight, or 30-60% by weight. Each possibility represents a distinct embodiment of the present invention.

[0040] In some embodiments, the composition contains phytofluene in amounts of 8-15% by weight, 10-20% by weight, 7-16% by weight, 12-19% by weight, 11-15% by weight, or 9-14% by weight of the composition. Each possibility represents a distinct embodiment of the present invention.

[0041] In some embodiments, the composition contains neuropolene in amounts of 4-15% by weight, 10-20% by weight, 5-16% by weight, 11-19% by weight, 7-15% by weight, 2-10% by weight, or 3-14% by weight of the composition. Each possibility represents a distinct embodiment of the present invention.

[0042] In some embodiments, the composition contains 10-20% by weight, 12-24% by weight, 15-25% by weight, 16-28% by weight, 16-27% by weight, or 14-23% by weight of ζ-carotene. Each possibility represents a distinct embodiment of the present invention.

[0043] In some embodiments, the weight / weight ratio of both phytoene:phytofluene and ζ-carotene is in the range of 2:1 (w / w) to 4:1 (w / w) or 2:1 (w / w) to 3:1 (w / w). Each possibility represents a distinct embodiment of the present invention.

[0044] In some embodiments, the composition contains trans-lycopene in amounts less than 10% by weight, less than 7% by weight, less than 5% by weight, less than 3% by weight, less than 2% by weight, or less than 1% by weight, or any value or range in between. Each possibility represents a separate embodiment of the present invention. In some embodiments, the composition contains trans-lycopene in amounts of 1-3% by weight, 1-5% by weight, 2-6% by weight, 0.5-4.5% by weight, 0.1-3% by weight, 0.6-4.8% by weight, or 2.5-4% by weight. Each possibility represents a separate embodiment of the present invention.

[0045] In some embodiments, the composition contains β-carotene in amounts less than 10% by weight, less than 7% by weight, less than 5% by weight, less than 3% by weight, less than 2% by weight, or less than 1% by weight, or any value or range in between. Each possibility represents a separate embodiment of the present invention. In some embodiments, the composition contains β-carotene in amounts of 1-3% by weight, 1-5% by weight, 2-6% by weight, 0.5-4.5% by weight, 0.1-3% by weight, 0.6-4.8% by weight, or 2.5-4% by weight. Each possibility represents a separate embodiment of the present invention.

[0046] In some embodiments, the composition comprises less than 5% by weight of trans-lycopene, less than 3% by weight of β-carotene, or a combination thereof.

[0047] In some embodiments, the composition further comprises tocopherol (e.g., vitamin E). In some embodiments, the composition contains tocopherol in amounts of 0.5-3% by weight, 1-5% by weight, 2-5.5% by weight, 4-6% by weight, 1.5-4.5% by weight, 3.5-5% by weight, or 2.5-4% by weight of the composition. Each possibility represents a distinct embodiment of the present invention.

[0048] Methods for determining the amount of plant nutrients such as carotenoids are common and will be apparent to those skilled in the art. Non-limiting examples of such methods include, but are not limited to, gas chromatography, liquid chromatography, and mass spectrometry.

[0049] In some embodiments, compositions are provided for use in reducing oxidative stress.

[0050] As used herein, the term “oxidative stress” refers to an imbalance between the action of reactive oxygen species (ROS) and the ability of a biological system (e.g., a cell, tissue, organ, object, or combination thereof) to eliminate (e.g., detoxify) such ROS and / or repair the damage caused thereby.

[0051] In some embodiments, oxidative stress is caused by or a result of radiation exposure. In some embodiments, the radiation includes UV / visible light wavelengths. In some embodiments, ultraviolet light is UVA, UVB, UVC, or any combination thereof. In some embodiments, the radiation includes sunlight.

[0052] In some embodiments, the radiation includes any radiation wavelength within the light spectrum. The term “light spectrum” as used herein refers to 10 -9 m~10 -3 It encompasses wavelengths in the range of m. In some embodiments, radiation wavelengths in the light spectrum include ultraviolet light, visible light, infrared light, or a combination thereof. In some embodiments, exposure to radiation includes exposure to sunlight.

[0053] As used herein, the term "ultraviolet (UV)" encompasses any wavelength in the UV region. In some embodiments, ultraviolet is UV. In some embodiments, ultraviolet is UVA, UVB, UVC, or any combination thereof.

[0054] In some embodiments, the composition is suitable for reducing oxidative stress caused by or arising from exposure to sunlight. In some embodiments, the composition is suitable for reducing oxidative stress caused by or arising from exposure to radiation with a continuous wavelength of 290 nm to 520 nm. In some embodiments, the composition is suitable for reducing oxidative stress caused by or arising from exposure to radiation with wavelengths including 420 nm to 450 nm, 430 nm to 455 nm, 440 nm to 450 nm, 440 nm to 460 nm, 440 nm to 470 nm, 450 nm to 465 nm, or 445 nm to 475 nm.

[0055] Methods for determining oxidative stress levels are common and will be apparent to those skilled in the art. Non-limiting examples of such methods are revisited by Katerji et al., 2019 (Oxidative Medicine and Cellular Longevity), and the ARE assay is illustrated below herein.

[0056] In some embodiments, the composition is an oral composition. In some embodiments, the composition is a pharmaceutical composition or a nutritional supplement composition. In some embodiments, the composition is a topical composition. In some embodiments, the composition comprises an acceptable carrier or excipient for pharmaceuticals or nutritional supplements.

[0057] In some embodiments, the oral composition is in the form of a soft gel capsule, tablet, two-piece capsule, or oral dispersible film (ODF). In some embodiments, the oral composition is in the form of a beverage, injection, gummy, or powder. In some embodiments, the oral composition is mixed or assimilated into a food product such as chocolate or ice cream.

[0058] In some embodiments, the topical composition is in the form of an ointment, cream, oil, or lotion.

[0059] In one embodiment, the composition of the present invention can be provided as an individual by itself. In one embodiment, the composition of the present invention can be provided as an individual as part of a pharmaceutical composition or nutritional supplement composition comprising an acceptable carrier for pharmaceuticals or nutritional supplements.

[0060] In one embodiment, “pharmaceutical composition” or “nutritional supplement composition” refers to a preparation of the composition described herein, comprising other chemical components such as physiologically suitable carriers and excipients. The purpose of the pharmaceutical composition or nutritional supplement composition is to facilitate the administration of the composition to an organism.

[0061] In some embodiments, a process is provided for producing a composition comprising 3 to 10% by weight of prolycopene and an acceptable carrier. In some embodiments, the process comprises extracting tangerine tomatoes as disclosed herein. In some embodiments, the composition of the present invention comprises a tangerine tomato extract produced by the process disclosed herein.

[0062] In one embodiment, the terms “physiologically acceptable carrier” and “pharmaceutically acceptable carrier,” used interchangeably, refer to a carrier or diluent that does not cause significant irritation to mammals and does not negate the biological activity and properties of the administered composition. Adjuvants are included in these terms.

[0063] In one embodiment, “excipient” refers to an inert substance added to the composition to further facilitate the administration of the active ingredient. In one embodiment, excipients include calcium carbonate, calcium phosphate, various sugars and various starches, cellulose derivatives, gelatin, vegetable oil and polyethylene glycol.

[0064] The techniques for drug formulation and administration are described in the latest edition of "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA, and the entire contents of that edition are incorporated herein by reference.

[0065] In one embodiment, preferred routes of administration include, for example, oral, rectal, transmucosal, transnasal, enteral, or parenteral delivery (including intramuscular, subcutaneous, and intrathecal injections, as well as subarachnoid, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections).

[0066] According to some embodiments, a method is provided for treating or preventing an oxidative stress-related condition or disease in a subject requiring treatment, comprising administering a therapeutically effective amount of the composition disclosed herein to the subject.

[0067] As used herein, “condition or disease associated with oxidative stress” encompasses any condition or disease (e.g., “pathological” condition) that impairs the health, survival, viability, functionality, or any combination thereof of the cells or objects in which it is contained, and this includes oxidative stress as part of its etiology, pathophysiology, or both.

[0068] Non-exclusive examples of conditions or diseases associated with oxidative stress include, but are not limited to, inflammation, cell proliferation-related disorders (e.g., cancer), and neurodegenerative diseases.

[0069] In some embodiments, administration includes oral administration.

[0070] As used herein, the terms “treatment” or “to treat” a disease, disorder, or condition encompass alleviation of at least one symptom, reduction of its severity, or inhibition of its progression. Treatment does not necessarily mean that the disease, disorder, or condition is completely cured. To be an effective treatment, the useful compositions herein only need to reduce the severity of a disease, disorder, or condition, or the severity of any symptom associated therewith, or to provide an improvement in the quality of life of the patient or subject.

[0071] As used herein, the term “prevention” of a disease, disorder, or condition encompasses delaying, preventing, suppressing, or inhibiting the onset of the disease, disorder, or condition. As used in accordance with the subject matter described herein, the term “prevention” relates to a preventive process in which the subject is exposed to the composition or formulation described herein before the induction or onset of the disease / disorder process. This can be done when an individual has a genetic lineage that shows a predisposition to the development of the disease / disorder being prevented. For example, this may apply to an individual whose ancestors show a predisposition to a particular type of disease, such as an inflammatory disease. The term “suppression” is used to describe a state in which the disease / disorder process has already begun, but the obvious symptoms of the condition are not yet apparent. Thus, an individual’s cells may have the disease / disorder, but the visible signs of that disease / disorder are not yet clinically apparent. In either case, the term “prevention” can be applied to encompass both prevention and suppression. Conversely, the term “treatment” refers to the clinical application of an active agent to combat an already existing condition in which its clinical symptoms are already apparent in the patient.

[0072] In some embodiments, prevention includes reducing the severity of the disease, delaying the onset of the disease, reducing the cumulative incidence of the disease, or any combination thereof.

[0073] In some embodiments, the method includes providing or selecting subjects who require prevention of conditions or diseases related to oxidative stress.

[0074] According to some embodiments, a method is provided for preventing an oxidative stress-related condition or disease in a subject in need, comprising (a) providing a subject at risk of developing an oxidative stress-related condition or disorder, and (b) administering a composition comprising a therapeutically effective amount of the composition in an amount of 1 to 15% by weight of prolycopene and an acceptable carrier to the subject.

[0075] As used herein, the terms “subject,” “individual,” “animal,” “patient,” or “mammal” refer to any subject, in particular a mammal subject for which treatment is desired, such as human.

[0076] Unless otherwise specified in this discussion, adjectives such as “substantially” and “about” modifying a state or relationship characterized by one or more features of one embodiment of the present invention are understood to mean that the state or characteristic is within the permissible limits for the implementation of the embodiment for the intended use. Unless otherwise specified, the word “or” in this specification and the claims shall be considered an inclusive “or” rather than an exclusive “or,” and shall indicate at least one or any combination of the items it connects.

[0077] Naturally, the terms “one (a)” and “one (an)” used above and elsewhere in this specification refer to “one or more of the enumerated components.” Unless otherwise specifically stated, it will be obvious to those skilled in the art that the singular use includes the plural. Accordingly, the terms “one (a),” “one (an),” and “at least one” are used synonymously in this application.

[0078] To better understand this instruction and in no way limit its scope, unless otherwise specified, all numbers and other numerical values ​​used herein and in the claims to represent quantities, percentages, or ratios should be understood to be modified in all cases by the term “approximately.” Therefore, unless otherwise stated, the numerical parameters described in the following details and in the attached claims are approximations that may vary depending on the desired characteristics to be obtained. At least each numerical parameter should be interpreted by taking into account at least the number of significant figures reported and by applying common rounding techniques.

[0079] In the specification and claims of this application, the verbs “comprise,” “include,” and “have,” as well as their conjugations, are used to indicate that one or more objects of a verb are not necessarily a complete list of components, elements, or parts of one or more subjects of that verb.

[0080] Other terms used herein are intended to be defined by their well-known meanings in the art.

[0081] Unless otherwise specified or evident from the context, the term “or” as used herein is understood to be inclusive.

[0082] Throughout this specification and the claims, the word “comprise” or variations such as “comprises” or “comprising” indicate the inclusion of any enumerated integer or group of integers, but not the exclusion of any other integer or group of integers.

[0083] The term "consists essentially of" or variations such as "consisting essentially of" as used throughout this specification and the claims indicate the inclusion of any enumerated integer or group of integers, and any inclusion of any enumerated integer or group of integers that does not substantially alter the basic or novel properties of the specified method, structure or composition.

[0084] As used herein, “comprises,” “comprising,” “containing,” and “having” can mean “includes” and “including,” and “consisting essentially of” or “consists essentially” also have the same meaning under U.S. patent law. This term is non-exclusive and acknowledges the existence of things not enumerated unless the basic or novel characteristics enumerated are altered by the existence of things not enumerated, but excludes embodiments of the prior art. In one embodiment, the terms “comprises,” “comprising,” and “having” are interchangeable with “consisting.”

[0085] Further objectives, advantages, and novel features of the present invention will become apparent to those skilled in the art by considering the following embodiments, which are not intended to be limiting. Furthermore, various embodiments and aspects of the present invention, detailed herein and claimed in the following claims, are experimentally supported in the following embodiments. [Examples]

[0086] In general, the nomenclature used herein and the laboratory procedures utilized in the present invention include chemical, molecular, biochemical, and cell biological techniques. Such techniques are fully described in the literature. For example, "Molecular Cloning: A Laboratory Manual," Sambrook et al., (1989); "Current Protocols in Molecular Biology," Volumes I-III, edited by Ausubel, RM (1994); "Cell Biology: A Laboratory Handbook," Volumes I-III, edited by Cellis, JE (1994); "The Organic Chemistry of Biological Pathways," by John McMurry and Tadhg Begley (Roberts and Company, 2005); "Organic Chemistry of Enzyme-Catalyzed Reactions," by Richard Silverman (Academic Press, 2002); "Organic Chemistry (6th edition)," by Leroy “Skip” G Wade; "Organic Chemistry," by TW Graham See the works of Solomons and Craig Fryhle.

[0087] material and method Preparation of extracts Tangerine tomatoes (TG) were extracted under the following conditions (extraction type: supercritical CO2 extraction (SCE), pressure: 360 bar, and temperature: 60°C). The amount of crude extract from the raw material was 3.4% (w / w).

[0088] LycoMato extracts were obtained by (1) SCE as described above, and (2) ethyl acetate extraction (EA) as previously described (International Publication No. 2010082205A1).

[0089] A non-limiting list of compounds identified within supercritical extraction (SCE), including their relative amounts, is provided below in this specification (Table 2). [Table 2]

[0090] Content of TG compared to LycoMato Table 3 shows a non-limiting example of a comparison of the active ingredients of TG (extract of the present invention) and Lyc-O-Mato 6% (both SCE and EA). [Table 3]

[0091] cell Normal human dermal fibroblasts (NHDF) were purchased from PromoCell (Heidelberg, Germany). These cells were grown in fibroblast growth medium 2 (PromoCell) according to the manufacturer's instructions.

[0092] T47D human breast cancer cells were grown in a humidified atmosphere containing 5% CO2 in DMEM containing penicillin (100 U / ml), streptomycin (0.1 mg / ml), nystatin (12.5 μg / ml), 10% FCS, and human recombinant insulin (6 μg / ml).

[0093] Transient transfection and ARE reporter gene assay Cells were transfected in 24-well plates using the jetPEI reagent (transfection reagent from Polyplus, Illkrich, France). NHDF primary human dermal fibroblasts or T47D human breast cancer cells (80,000 cells per well) were transfected with 0.2 μg of 4×ARE reporter construct and 0.1 μg of normalized plasmid. Cells were seeded in culture medium containing 3% fetal bovine serum (FCS). The following day, the cells were washed once with appropriate culture medium, and then 0.45 mL of medium and 50 μl of DNA mixed with jetPEI were added. The cells were then incubated at 37°C for 4–6 hours. The used medium was replaced with fresh medium supplemented with 3% FCS containing the test compound, and the cells were incubated for a further 16–20 hours. ARE / Nrf2 reporter activity was measured in cell extracts and normalized to sea urchin luciferase using a dual luciferase reporter assay system (Promega, Madison, Wisconsin, USA) according to the manufacturer's instructions. The reporter construct used was a 4× ARE reporter construct. The sea urchin luciferase (P-RL-null) expression vector served as the internal transfection standard (Promega, Madison, Wisconsin, USA).

[0094] Example 1: Tangerine tomato extract increases the antioxidant stress response in cells. The induction of ARE by TG extract in NHDF dermal fibroblasts was found to be approximately 3 times higher than that of Lyc-O-Mato when compared to the same concentration of lycopene as a reference (Figure 1). Antioxidant response element (ARE) activity is considerably low in this type of cell, so we compared the activity of the two extracts in T47D breast cancer cells, which exhibit higher activity. The induction of ARE in this type of cell was found to be dose-dependent, with induction by TG extract being approximately 60 times greater at 20 μM lycopene and approximately 160 times greater at 40 μM lycopene compared to the Lyc-O-Mato control.

[0095] The induction of ARE, a marker of the cellular antioxidant defense system, was approximately 20 times higher when using the TG extract than when using the Lyc-O-Mato control. Even when this comparison was performed using the total amount of carotenoids, which was 6 times higher in the TG extract, the activity was still approximately 3 times higher in the TG extract, suggesting that the antioxidant protection of cells by the TG extract in this experimental setup is superior to that by Lyc-O-Mato.

[0096] Example 2: TG extract has high bioavailability. Pharmacokinetic experiments will be conducted. A crossover supplementation will be performed using either TG or LycoMato capsules. Two groups will be compared: (1) TG Tomato capsules containing 15 mg of lycopene (approximately 1:2 (w / w) trans-lycopene:tetra-cis-lycopene (pro)), 41.5 mg of phytoene, 13.3 mg of phytofluene, 20.4 mg of ζ-carotene, 0.6 mg of β-carotene, and 5.5 mg of tocopherol; and (2) Lyco-Mato capsules containing 15 mg of trans-lycopene, 6.5 mg of phytoene, 1.5 mg of phytofluene, 0.3 mg of ζ-carotene, 0.45 mg of β-carotene, and 5.1 mg of tocopherol.

[0097] Experimental design: [Table 4] [Table 5]

[0098] The plasma concentrations of lycopene, phytoene, phytofluene, ζ-carotene, and β-carotene will be determined.

[0099] Although the present invention has been described in particular, those skilled in the art will understand that many modifications and alterations are possible. Therefore, the present invention should not be construed as being limited to the embodiments described herein, and the scope and concept of the present invention will be more readily understood by referring to the following claims.

Claims

[Claim 1] A composition comprising 1 to 15% by weight of prolycopene and an acceptable carrier.

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