US20220040251A1

US20220040251A1 - Compositions and methods for nutritional supplements

Info

Publication number: US20220040251A1
Application number: US17/508,543
Authority: US
Inventors: Hartley Pond
Original assignee: DailyColors Health Inc
Current assignee: DailyColors Health Inc
Priority date: 2020-02-05
Filing date: 2021-10-22
Publication date: 2022-02-10
Anticipated expiration: 2041-02-04
Also published as: JP2023512778A; KR20220141310A; AU2021216555A1; US11202816B1; US20220125871A1; CN115335068A; WO2021158825A1; CA3166761A1; US11065295B1; EP4100033A1; US11752188B2; US20210244786A1; US11246903B1; EP4100033A4

Abstract

Compositions and methods are presented in which a plurality of chemically distinct polyphenols inhibit multiple enzymes in pathways associated with health and healthy ageing. Preferred compositions are derived from colored plant materials that are commonly found in the Mediterranean diet and provide the biochemical basis for the health benefits of the Mediterranean diet. Notably, the enzyme inhibition observed with the combined polyphenols was synergistic with respect to not one but a significant number of enzymes in the pathways associated with health and healthy ageing, thus providing an amplified desirable effect.

Description

This application claims priority to our co-pending U.S. patent application with the Ser. No. 17/360,261, which was filed Jun. 28, 2021, which claims priority to U.S. Pat. No. 11,065,295, filed Feb. 4, 2021, which claims priority to U.S. Provisional Patent Applications with the Ser. No. 62/970,615, filed Feb. 5, 2020, Ser. No. 63/010,183, filed Apr. 15, 2020, and Ser. No. 63/086,207, filed Oct. 1, 2020, each of which is incorporated by reference herein.

FIELD OF THE INVENTION

The field of the invention is compositions and methods for nutritional supplements, especially as it relates polyphenols and polyphenol mixtures commonly associated with a diet rich in fruits and vegetables and their use in conditions, disorders, and diseases associated with various enzymes inhibited by such polyphenol mixtures.

BACKGROUND OF THE INVENTION

The background description includes information that may be useful in understanding the present disclosure. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
All publications and patent applications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.
There is a considerable variety of vitamins and other isolated nutritional compounds, and alleged benefits of such compounds include, among numerous other effects, immune support, anti-inflammatory effects, anti-ageing effects, cardiac support, and digestive support. Unfortunately, there is only a rather small body of evidence that substantiates some aspects of these alleged benefits when these vitamins and other isolated nutritional compounds are ingested. Similarly, where the nutritional supplement is an extract or powdered form of a plant part, various benefits are advertised, but actual benefits are often poorly or even not at all proven. Moreover, isolated nutritional compounds as well as individual plant extracts and concentrates are generally not reflective of a healthy diet.
Notably, there are certain geographic and ethnographic diet types that are associated with overall health, longevity, and/or physical resilience, and such beneficial effects are indeed well documented and substantiated. For example, the Mediterranean diet is typically associated with lower cardiovascular risk factors (see e.g., Nutrients 2018, 10, 379; doi:10.3390/nu10030379), lower inflammatory and metabolic biomarkers, a reduction in risk of Alzheimer's disease (see e.g., J Alzheimers Dis. 2010; 22(2): 483-492), and with a reduction in certain inflammatory markers (see e.g., Nutrients 2018, 10, 62; doi:10.3390/nu10010062). One common ingredient class found in such diets are polyphenols, and various studies have been published regarding specific benefits of individual dietary polyphenols (see e.g., Inhibitory Properties of Phenolic Compounds Against Enzymes Linked with Human Diseases: URL:dx.doi.org/10.5772/66844), and selected colored polyphenols (see e.g., Annu. Rev. Food Sci. Technol. 2020. 11:10.1-10.38). However, due to the complexity and large number of chemically distinct polyphenols, many studies only focus on single polyphenols and particular biochemical effects of such compounds or provide general epidemiological information without more detailed molecular characterization of the diets.
In an effort to supplement a diet with multiple polyphenols, various supplements are known. For example, Vital Reds (by Gundry M D) provides a commercially available concentrated polyphenol powder blend from a number of red colored plant materials to increase energy and improve digestion. Such blend advantageously includes a variety of chemically distinct polyphenols. However, the selection of plant materials used as a source of polyphenols is not reflective of common dietary intake. Similarly, Oxxynea by Fytexia, a commercially available mixture of grape, olive, pomegranate, green tea, grapefruit, bilberry, and orange extracts is offered as an antioxidant formulation to protect cells from oxidative stress (see e.g., Oxxynea by Fytexia). While beneficial to reduce oxidative stress, the source ingredients for such antioxidant formulation are once more not reflective of a common dietary intake. Surprisingly, despite the numerous beneficial components found in various dietary supplements, there is no supplement that is expected to provide the various benefits of a Mediterranean diet, and particularly the benefits of colored polyphenolic components in Mediterranean diet.
Thus, even though various nutritional supplements are known in the art, all or almost all of them suffer from various disadvantages. Consequently, there is a need to provide improved compositions and methods for nutritional supplements, and especially beneficial/synergistic combinations of polyphenols known to be associated with healthy diets such as the Mediterranean diet.

SUMMARY OF THE INVENTION

The inventor has now discovered various compositions and methods for specific combinations of polyphenols and/or polyphenol-rich materials (e.g., extracts and powders) commonly found in food items of the Mediterranean diet that exhibited, when combined, numerous benefits associated with the benefits of the Mediterranean diet. Indeed, the compositions and methods disclosed herein had substantial, and in some cases significant synergistic effect on a variety of molecular biomarkers associated with the benefits of the Mediterranean diet such as markers for ageing, senescence, inflammation, immune function, NAD/energy metabolism, and the gut microbiome.
In one aspect of the inventive subject matter, the inventor contemplates a nutritional composition that comprises a nutritionally acceptable carrier in combination with a plurality of chemically distinct polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color. Preferably, the chemically distinct polyphenols from the plant materials are present as a synergistic combination with respect to inhibition of at least one biochemical marker selected from the group consisting of BACE1, CD38, CD73, CDK5, JAK1, JAK2, and JAK3.
For example, in some embodiments the red colored plant materials comprise at least one (or two, or three, or all of) of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials comprise at least one (or two, or three, or all of) of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials comprise at least one (or two, or three, or all of) of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials comprise at least one (or two, or three, or all of) of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
In further aspect of the inventive subject matter, the chemically distinct polyphenols further inhibit at least one additional biochemical marker selected from the group consisting of ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S. Moreover, contemplated compositions may further inhibit Keap/Nrf2 binding and/or ACE2/Spike binding.
Most typically, but not necessarily, the composition is formulated in single dosage units for oral administration (e.g., each containing between 50 and 1,000 mg of the composition), which may be formulated as a capsule, a gummy, or a powder. Where desired, the composition may further comprise a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic.
Therefore, in another aspect of the inventive subject matter, the inventor contemplates a nutritional composition that includes a nutritionally acceptable carrier in combination with a plurality of chemically distinct polyphenol-containing plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color. Most preferably, the red colored plant materials comprise an apple extract, a pomegranate extract, a tomato powder, and a beet root powder; the green colored plant materials comprise an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder; the orange-yellow colored plant materials comprise an onion extract, a ginger extract, a grapefruit extract, and a carrot powder; and the purple-blue colored plant materials comprise a grape extract, a blueberry extract, a currant powder, and an elderberry powder. For example, the apple extract, the pomegranate extract, the olive extract, the rosemary extract, the green coffee bean extract, the onion extract, the ginger extract, the grapefruit extract, the grape extract, and the blueberry extract may be ethanol extracts or ethanol/water extracts.
In such compositions the combination of plant materials is a synergistic combination with respect to inhibition of at least one biochemical marker selected from the group consisting of BACE1, CD38, CD73, CDK5, JAK1, JAK2, and JAK3, and especially with respect to inhibition of BACE1, CD38, and CD73. Preferred compositions further inhibit at least one additional biochemical marker selected from the group consisting of ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S, and may also inhibit Keap/Nrf2 binding and/or ACE2/Spike binding.
As before, it is preferred (but not needed) that the composition is formulated in single dosage units for oral administration. Typically, the dosage unit contains between 50 and 1,000 mg of the composition, and is formulated as a capsule, a gummy, or a powder. Where desired, contemplated compositions may further comprise a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic.
Among other uses, contemplated compositions will be effective to treat and/or reduce a symptom of inflammatory condition, a cardiovascular condition, a neurological condition, a metabolic condition, and a cancer.
Therefore, the inventor also contemplates a method of supporting health of an individual that comprises a step of administering the compositions presented herein. For example, the composition may be administered to thereby provide immune support, metabolic support, support longevity, support central nervous system (CNS) function, reduce an inflammatory response, reduce effects of cardiovascular disease, and reduce the rate of amyloid beta plaque formation.
In further examples of such methods, the composition is orally administered over at least 30 days, typically at a daily dose of between 50 and 1,000 mg. As noted earlier, contemplated compositions may further comprise a step of co-administering a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic (which may be performed in the same dosage unit or individually).
In another aspect of the inventive subject matter the inventor also contemplates a method of reducing an NAD+ decrease (e.g., age-related NAD+ decrease) in an individual that includes a step of administering to the individual a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color wherein the combination synergistically inhibits CD38. In some embodiments, the polyphenols are provided in from of the plant materials.
Preferably, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
In a further aspect of the inventive subject matter the inventor contemplates a method of supporting longevity of an individual that includes a step of administering to the individual a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color wherein the combination synergistically inhibits CD73. In some embodiments, the polyphenols are provided in from of the plant materials.
Preferably, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
In yet another aspect of the inventive subject matter the inventor contemplates a method of supporting cognitive function of an individual that includes a step of administering a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color wherein the combination synergistically inhibits BACE1. In some embodiments, the polyphenols are provided in from of the plant materials, and administration increases immune function to thereby support longevity and/or reduces the rate of amyloid beta plaque formation.
Preferably, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
In still another aspect of the inventive subject matter the inventor contemplates a method of supporting central nervous system (CNS) function in an individual that includes a step of administering a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color wherein the combination synergistically inhibits CDK5. In some embodiments, the polyphenols are provided in from of the plant materials, and administration reduces age-related cognitive decline.
Preferably, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
In a further aspect of the inventive subject matter the inventor contemplates a method of supporting immune function in an individual that includes a step of administering a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color wherein the combination synergistically inhibits at least one of JAK1, JAK2, and JAK3. In some embodiments, the polyphenols are provided in from of the plant materials, and administration reduces a symptom of rheumatoid arthritis, psoriasis, or inflammatory bowel disease.
Preferably, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
Additionally, the inventor also contemplates a method of inhibiting at least one of BACE1, CD38, CD73, CDK5, JAK1, JAK2, and JAK3 that includes a step of contacting at least one of the BACE1, the CD38, the CD73, the CDK5, the JAK1, the JAK2, and the JAK3 with a plurality of chemically distinct polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color, wherein the chemically distinct polyphenols from the plant materials are a synergistic combination with respect to inhibition of at least one biochemical marker selected from the group consisting of the BACE1, the CD38, the CD73, the CDK5, the JAK1, the JAK2, and the JAK3. In some embodiments, the polyphenols are provided in from of the plant materials.
For example, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
Advantageously, the step of contacting is performed in vivo (e.g., oral administration to a mammal), and administration of the plurality of chemically distinct polyphenols provides immune support, metabolic support, support longevity, supports central nervous system (CNS) function, reduces an inflammatory response, reduces effects of cardiovascular disease, and/or reduces the rate of amyloid beta plaque formation.
Viewed from a different perspective, the inventor also contemplates a method of treating a condition that is associated with activity of at least one of BACE1, CD38, CD73, CDK5, JAK1, JAK2, JAK3, ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S in a mammal. Such method will typically include a step of administering to the mammal a plurality of chemically distinct polyphenols in an amount effective to inhibit at least one of the BACE1, the CD38, the CD73, the CDK5, the JAK1, the JAK2, the JAK3, the ARG-1, the ARG-2, the SIRT-1, the CD39, the IDO1, the IDO2, the NAMPT, the PCSK9, the CD47, and the Cathepsin S. In preferred embodiments, the chemically distinct polyphenols synergistically inhibit BACE1, CD38, CD73, CDK5, JAK1, JAK2, and/or JAK3.
For example, the condition may be an inflammatory condition, a cardiovascular condition, a neurological condition, a metabolic condition, and/or a cancer. Where desired, the plurality of chemically distinct polyphenols are from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color. For example, the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder. It should also be appreciated that the polyphenols may be provided in from of the plant materials.
Moreover, it is contemplated that the plurality of chemically distinct polyphenols may be orally administered to the mammal, typically at a daily dosage of between 50 and 1,000 mg. Where desired, such methods may further comprise a step of co-administering to the mammal a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic.
In still further contemplated embodiments, the chemically distinct polyphenols are administered in an amount effective to inhibit at least three (or at least five or at least ten or all) of the BACE1, the CD38, the CD73, the CDK5, the JAK1, the JAK2, the JAK3, the ARG-1, the ARG-2, the SIRT-1, the CD39, the IDO1, the IDO2, the NAMPT, the PCSK9, the CD47, and the Cathepsin S.
Therefore, and viewed from a different perspective, the inventor also contemplates the use of a plurality of chemically distinct polyphenols to support healthy ageing. Moreover, it is contemplated that the plurality of chemically distinct polyphenols in such use further inhibit SIRT-1, IDO1, IDO2, NAMPT, PCSK9, CD47, Keap/Nrf2 binding, and/or ACE2/Spike binding, and/or that the chemically distinct polyphenols synergistically inhibit the BACE1, the CD38, the CD73, the CDK5, the JAK1, the JAK2, and/or the JAK3.
As before, it is preferred that the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.
Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments, along with the accompanying drawing figures in which like numerals represent like components.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a graph depicting exemplary results for ARG-1 inhibition using a composition according to the inventive subject matter.

FIG. 2 is a graph depicting exemplary results for ARG-2 inhibition using a composition according to the inventive subject matter.

FIG. 3 is a graph depicting exemplary results for SIRT1 inhibition using a composition according to the inventive subject matter.

FIG. 4 is a graph depicting exemplary results for Keap1-Nrf2 binding inhibition using a composition according to the inventive subject matter.

FIG. 5 is a graph depicting exemplary results for ACE2-Spike S1 binding inhibition using a composition according to the inventive subject matter.

FIG. 6 is a graph depicting exemplary results for ACE2-Spike S1 binding inhibition using a multivitamin composition.

FIG. 7 is a graph depicting exemplary results for ACE2-Spike S1 binding inhibition using various further compositions according to the inventive subject matter

FIG. 8 is a graph depicting exemplary results for BACE1 inhibition using a composition according to the inventive subject matter.

FIG. 9 is a graph depicting exemplary results for BACE1 inhibition using various further compositions according to the inventive subject matter and a multivitamin composition.

FIG. 10 is a graph depicting exemplary results for Cathepsin S inhibition using a composition according to the inventive subject matter.

FIG. 11 is a graph depicting exemplary results for Cathepsin S inhibition using various further compositions according to the inventive subject matter.

FIG. 12 is a graph depicting exemplary results for Cathepsin S inhibition using a composition according to the inventive subject matter and a multivitamin composition.

FIG. 13 is a graph depicting exemplary results for CDK5/p25 binding inhibition using a composition according to the inventive subject matter.

FIG. 14 is a graph depicting exemplary results for CDK5/p25 binding inhibition using various further compositions according to the inventive subject matter and a multivitamin composition.

FIG. 15 is a graph depicting exemplary results for IDO1 inhibition using a composition according to the inventive subject matter.

FIG. 16 is a graph depicting exemplary results for IDO2 inhibition using a composition according to the inventive subject matter.

FIG. 17 is a graph depicting exemplary results for NAMPT inhibition using a composition according to the inventive subject matter.

FIG. 18 is a graph depicting exemplary results for PCSK9:LDLR binding inhibition using a composition according to the inventive subject matter.

FIG. 19 is a graph depicting exemplary results for CD47 inhibition using a composition according to the inventive subject matter.

FIG. 20 is a graph depicting exemplary results for CD38 inhibition using a composition according to the inventive subject matter.

FIG. 21 is a graph depicting exemplary results for CD38 inhibition using further compositions according to the inventive subject matter.

FIG. 22 is a graph depicting exemplary results for CD38 inhibition using known compositions containing nicotinamide riboside.

FIG. 23 is a graph depicting exemplary results for CD38 inhibition using a composition according to the inventive subject matter and a known multivitamin composition.

FIG. 24 is a graph depicting exemplary results for JAK1 inhibition using a composition according to the inventive subject matter.

FIG. 25 is a graph depicting exemplary results for JAK2 inhibition using a composition according to the inventive subject matter.

FIG. 26 is a graph depicting exemplary results for JAK3 inhibition using a composition according to the inventive subject matter.

FIG. 27 is a graph depicting exemplary results for CD39 inhibition using a composition according to the inventive subject matter.

FIG. 28 is a graph depicting exemplary results for CD39 inhibition using a composition at low concentrations according to the inventive subject matter.

FIG. 29 is a graph depicting exemplary results for CD39 inhibition using further compositions according to the inventive subject matter.

FIG. 30 is a graph depicting exemplary results for CD39 inhibition using selected compositions according to the inventive subject matter.

FIG. 31 is a graph depicting exemplary results for CD39 inhibition using a composition according to the inventive subject matter and a known multivitamin composition.

FIG. 32 is a graph depicting exemplary results for CD73 inhibition using a composition according to the inventive subject matter.

FIG. 33 is a graph depicting exemplary results for CD73 inhibition using a composition at low concentrations according to the inventive subject matter.

FIG. 34 is a graph depicting exemplary results for CD73 inhibition using further compositions according to the inventive subject matter

FIG. 35 is a graph depicting exemplary results for CD73 inhibition using a composition according to the inventive subject matter and a known multivitamin composition.

DETAILED DESCRIPTION

The inventor has discovered that specific combinations of polyphenol-containing materials (and polyphenols found therein) strongly modulated numerous biomarkers associated with the beneficial effects of the Mediterranean diet. In view of these findings, the inventor therefore contemplates various compositions for nutritional supplements and other nutritional products, compositions, and uses in medicinal food, and even use in medicine.
Most notably, the compositions presented herein had substantial and synergistic effects on a number of biomarkers associated with proper immune and CNS function, effective cellular metabolism, and longevity, and showed further significant effect on additional biomarkers associated with inflammatory responses, adverse effects of cardiovascular disease, and/or amyloid beta plaque formation. For example, the compositions and methods presented herein were demonstrated to have significant and beneficial effects on multiple enzymatic targets that are involved with numerous aspects of health and healthy ageing such as ARG-1, ARG-2, SIRT1, BACE1, Cathepsin S, CDK5, IDO1, IDO2, NAMPT, PCSK9, CD47, CD38, JAK1, JAK2, JAK3, CD39, and CD73, Keap/Nrf2. Viewed from a different perspective, it should be appreciated that the compositions presented herein are useful to beneficially affect multiple pathways associated with health and healthy ageing via inhibition of key signaling components and/or enzymes in these pathways. Remarkably, the observed modulation of these biomarkers using the compositions presented herein paralleled the profile of biomarkers in individuals that adhered to the Mediterranean diet and individuals with significant longevity.
For example, arginase 1 (ARG1) and arginase 2 (ARG2) are key enzymes in the urea cycle, cleaving L-arginine to form urea and L-omithine. The urea cycle provides protection against excess ammonia, while L-ornithine is required for cell proliferation, collagen formation, and other important physiological functions. Notably, increases in arginase activity in mammals have been linked to dysfunction and pathologies of the cardiovascular system, kidney, and central nervous system, and also to dysfunction of the immune system and development of cancer. Two important aspects of the excessive activity of arginase may be involved in diseases. First, overly active arginase can reduce the supply of L-arginine needed for the production of nitric oxide (NO) by NO synthase. Second, excessive quantities of L-ornithine can lead to structural problems in the vasculature, neuronal toxicity, and abnormal growth of tumor cells (see e.g., Physiol Rev 98: 641-665, 2018). Furthermore, studies have demonstrated that increased formation of reactive oxygen species and key inflammatory mediators promote a pathological elevation of arginase activity. As such, inhibition of ARG1 and ARG2 is thought to beneficially counteract the adverse effects of arginase overactivity. As is shown on more detail below, contemplated compositions were effective in inhibiting ARG1 and ARG2 activity even at relatively low concentrations.
In another example, sirtuin1 (SIRT1) is a nuclear enzyme that deacetylates transcription factors that contribute to cellular regulation, and particularly to reaction to stressors. For example, SIRT1 deacetylates members of the PGC1-alpha/ERR-alpha complex, which are critical metabolic regulatory transcription factors, and deacetylates/deactivates the p53 protein, which is a key factor in many neoplastic diseases. SIRT1 was also demonstrated to play a role in activating T helper 17 cells, which contribute to autoimmune disease. As is shown in more detail below, contemplated compositions were effective in mildly inhibiting SIRT1 activity at relatively moderate concentrations.
In yet another example, beta-secretase 1 (BACE1) has been shown to be essential for the generation of β-amyloid in Alzheimer's disease and has also been reported to be associated with cognitive decline and decline in central nervous system (CNS) function (see e.g., Molecules. 2017 Oct. 13; 22(10):1723). Indeed, β-amyloid accumulation is a hallmark of ageing, and as such inhibitory compounds are thought to beneficially decelerate or even stop β-amyloid accumulation and as such preserve or maintain cognitive abilities and CNS function. As is described in more detail below, contemplated compositions have shown remarkable, strong, and even synergistic effect with regard to BACE 1 inhibition.
In yet another example, higher levels of Cathepsin S have been reported to be associated with higher risk of mortality and were in some studies also associated with higher risk of cardiovascular mortality (e.g., JAMA, Sep. 14, 2011—Vol 306, No. 10 1113). Other experimental studies have suggested that cathepsin S activity is involved in the development of cardiovascular disease via promotion of atherosclerotic plaques and destabilization of advanced plaques. Moreover, cathepsin S activity was also implicated in the development of cancer via stimulation of cancer cell migration and tumor angiogenesis, and higher levels of Cathepsin S activity was reported to be correlated with ageing of the brain. As such, reduced levels of Cathepsin S activity are believed to beneficially counteract these risks. Notably, contemplated compositions have shown remarkable and strong inhibitory effect with regard to BACE 1 inhibition.
Likewise, cyclin-dependent kinase 5 (CDK5) has been found to be essential to proper CNS function and its role in peripheral tissue and disease is growing. For example, acute CDK5 inhibition has a high potential therapeutic value to prevent neuronal injury in the events of stroke or brain injury, or during high-risk surgeries, such as neurovascular or cardiovascular surgeries, or potentially during prolonged, complicated labors. Pharmacological inhibition of CDK5 has been shown to protect neurons under a range of different stressful conditions and ageing (see e.g., Curr Pharm Des. 2016; 22(5): 527-534). Once more, contemplated compositions have shown remarkable, strong, and synergistic inhibitory effect with regard to CDK5 inhibition.
In still other example, Janus kinases 1, 2, and 3 (JAK1, JAK2, JAK3) are implicated in the regulation of cell cycle and cancer, presumably via NF-kB activation (see e.g., Cells 2020, 9, 1451). Moreover, small-molecule drugs that inhibit Janus kinases (JAK1-3) were essential signaling mediators downstream of many proinflammatory cytokines and have gained traction as safe and efficacious options for immune-mediated disorders. Not surprisingly, inhibition of JAKs has emerged as leading potential treatment of inflammation-driven pathologies like rheumatoid arthritis, psoriasis, and inflammatory bowel disease, as well as cardiovascular disease. Notably, contemplated compositions have shown remarkable, strong, and synergistic inhibitory effect with regard to JAK1, JAK2, and JAK3 inhibition (especially at high concentrations).
In still further examples, inhibition of indoleamine 2,3-dioxygenase-1 (IDO1) has emerged as a new treatment strategy for immune support, and particularly in the reversion of tumor-mediated immune suppression (see e.g., Cancer Res. 2017 Dec. 15; 77(24): 6795-6811). Similarly, indoleamine 2,3-dioxygenase-2 (IDO2) has been implicated more recently in cancer, inflammation and immune control, and significant resources have been expended to identify IDO2 specific inhibitors. Increases in IDO1 and IDO2 activity have also been observed as a function of ageing (e.g., Immunity & Ageing 2011, 8:9), along with age related increase in inflammation, autoimmune disorders and malignancies. Remarkably, and as is shown in more detail below, contemplated compositions have shown strong inhibitory effect with regard to IDO1 and IDO2 inhibition.
With respect to energy metabolism, NAMPT and CD38 are known to modulate NAD+ in cells and as such are thought to be essential in maintaining and supporting of cellular energy and proper metabolic function. Indeed, overactivity of CD38 has been reported to increase with senescence (see e.g., Biochem Biophys Res Commun. 2019 May 28; 513(2): 486-493), cancer and ageing diseases (e.g., Trends Pharmacol Sci. 2018 April; 39(4): 424-436), immunomodulation and metabolic diseases (see e.g., FIMMU May 2019, Vol. 10, Article 1187). On that backdrop, inhibition of CD38 is believed to reduce or even prevent diseases associated with CD38 overactivity. On the other hand, nicotinamide phosphoribosyltransferase (NAMPT) mediates the rate-limiting step of the NAD salvage pathway that maintains cellular bioenergetics and provides a necessary substrate for functions essential to cells, and especially rapidly proliferating cancer cells. Once more, contemplated compositions have shown strong and synergistic inhibitory effect with regard to CD38 inhibition, and strong inhibition with respect to NAMPT activity.
Moreover, overactivity of CD39 and CD73 have been reported to contribute to immune suppression, suppression of checkpoint inhibition in tumors, and other aspect of immune regulation (see e.g., FIMMU June 2017, Volume 8, Article 727; Immunol Rev. 2017 March; 276(1): 121-144). In addition, centenarians showed a significantly lower expression of CD39 and CD73 as compared to younger individuals (here: octogenarians), suggesting that the levels of CD39 as well as CD73 mRNA could be a hallmark of successful human ageing. Viewed from a different perspective, aging is associated with a decline in immune function and so contributes to the increased susceptibility to infectious diseases and higher incidence of malignant disease. Therefore, lower levels of CD39 activity are thought to be directly associated with healthy aging and longevity. Remarkably, contemplated compositions had a significant and synergistic effect in the inhibition of CD73 and a profound inhibitory effect on the activity of CD39 as is shown in more detail below. Likewise, CD47 inhibition was shown to lead to stimulation of phagocytosis of cancer cells and CD47 blockade not only enhanced the function of innate immune cells but also linked to adaptive immune responses (see e.g., Cell Reports 31, 107494 Apr. 14, 2020). Notably, contemplated compositions had a significant inhibitory effect in the inhibition of CD47 as is shown in further detail below.
In yet other examples, proprotein convertase subtilisin/kexin type 9 (PCSK9) has been reported as a contributor to plasma cholesterol levels, and inhibitors targeting PCSK9 have reduced plasma cholesterol in human. Once more, and as shown in more detail below, contemplated compositions had a significant and synergistic effect in the inhibition of PCSK9.
Additionally, the Keap1-Nrf2 pathway is a major regulator of cytoprotective responses to endogenous and exogenous stresses caused by reactive oxygen species (ROS) and Nrf2 activates expression of a variety of genes encoding stress response proteins. Inhibitors of Keap1-Nrf2 binding are therefore thought to increase expression of stress response related proteins. Remarkably, and as shown in more detail below, contemplated compositions had a significant and effect in the inhibition of Keap1-Nrf2 binding (and as such increase availability of Nrf2).
Finally, the inventor also discovered that contemplated compositions also had inhibitory effect in ACE2-Spike protein binding, which is implicated in viral propagation of corona viruses, and especially Sars-CoV2. As will be readily appreciated, contemplated compositions may therefore provide at least some protective effect against corona viruses, and especially Sars-CoV2.
Based on his extensive research, the inventor has now discovered that specific blends of selected plant materials common in the Mediterranean diet can be prepared that mimic the benefits of the Mediterranean diet as evidenced in the modulation of biomarkers. Preferably, such blends are combinations of colored plant materials that belong to a number (e.g., at least two, at least three, or at least four) of different color groups, and particularly plant materials having a red color, green color, orange/yellow color, and/or purple/blue color. For example, in one embodiment of contemplated compositions, polyphenol-containing products/extracts were obtained from red colored source materials that included an apple extract, a pomegranate extract, tomato powder, and beet root; from green colored source materials that included an olive extract, rosemary extract, green coffee bean extract, and kale; from orange/yellow colored source materials that included an onion extract, a ginger extract, a grapefruit extract, and carrot; and from purple/blue colored source materials that included a grape extract, a blueberry extract, currant, and elderberry, and the particular ingredients and proportions are described in more detail below. Viewed from a different perspective, contemplated compositions will therefore include a large number of polyphenols that below to at least two, or at least three, or at least four different polyphenolic classes, including organic acids, phenolics, flavonols, flavanols, anthocyanins, chlorogenic acids, betacyanins, etc. As will be readily appreciated, the particular choice of a plant material will depend on the desired (polyphenolic) component in the plant material and its effect on a particular biological system and/or signaling pathway.
Of course, it should also be appreciated that the plant materials may be provided in various forms, including whole plant materials or portions thereof (e.g., root, fruit, leaves, etc.) in fresh or dried form, juices or macerates from plant materials or portions thereof in fresh or dried form, and aqueous or aqueous/alcoholic extracts and chromatographic fractions of the aforementioned plant materials. Still further, it should be noted that one or more polyphenols of the plant materials may even be provided as purified (natural isolated or synthetic) chemical entities, typically with a chemical purity of at least 90%, or at least 95%, or at least 98%, or at least 99%. However, it should be recognized that in most embodiments the plant materials will be complex mixtures to provide a combination of desired biological effects on a number of distinct molecular entities (e.g., enzymes, receptors, ion channels) where at least some of the biological effects (e.g., at least one, or at least two, or at least three, etc.) are synergistic. Moreover, it is contemplated that the biological effects on the particular molecular entities will also be complementary in biological function. Therefore, and based on the testing and desired targets as described in more detail below, it should be appreciated that the compositions of the inventive subject matter may be formulated to meet a particular need. However, in especially preferred aspects contemplated compositions will inhibit multiple targets (e.g., at least two, at least three, at least four, etc.) in multiple and distinct (e.g., at least two, at least three, at least four, etc.) signaling pathways.
Consequently, and viewed from a different perspective, it should be appreciated that the mechanism of action of contemplated compositions is not limited to a single specific function (e.g., antioxidant) or limited to a specific chemical category (e.g., vitamins), but in fact complementarily and synergistically provides multiple biological activities across distinct metabolic and signaling pathways. As such, contemplated compositions and methods target a variety of biological systems, including energy metabolism, immune function, neural and CNS function, cardiac function, inflammation, etc. Notably, and as is described in more detail below, the compositions contemplated herein also affected a number of biomarkers associated with longevity (e.g., in blue zone populations such as Mediterranean population, Okinawan population, etc.) In addition, it is contemplated that the plant materials will also provide a variety of micro-nutrients to assist or complement the functions of the polyphenols and other colored pigments present in the compositions.
Consequently, it should be appreciated that the compositions contemplated herein may be advantageously used as a stand-alone product to support various aspects of health and healthy ageing such as support of proper immune function, support to reduce inflammation, support of normal NAD+ levels, support of cardiac health. In this context, it should be noted that the term “support” when used in conjunction with a physiological function or condition is intended to mean prevent decline of one or more components or activities of the component(s) associated with the physiological function or condition, at least partially reverse decline of one or more components or activities of the component(s) associated with the physiological function or condition, maintain normal function of one or more components or activities of the component(s) associated with the physiological function or condition, prevent abnormal overactivity (or over-expression) of one or more components associated with the physiological function or condition, and/or at least partially reverse abnormal overactivity (or over-expression) of one or more components associated with the physiological function or condition. Alternatively, the compositions contemplated herein may also be combined with other nutritional supplements and/or vitamins to provide beneficial effects otherwise not obtainable with such supplements or vitamins. In this context, it should be appreciated that most, if not all of the biomarkers tested herein were not substantially modulated by multivitamin compositions as is shown in more detail below. Thus, it should be recognized that the compositions resented herein present a new and different class of health support with a variety of beneficial effects that reach beyond the benefits of a multivitamin formulation.
In further contemplated aspects of the inventive subject matter it should be appreciated that the compositions presented herein may be formulated in a variety of forms, and particularly preferred formulations include those in combination with a nutritionally or pharmaceutically acceptable carrier, most preferably for oral administration (however, parenteral administration is also expressly contemplated). Therefore, contemplated compositions can be formulated as solid or a liquid product. For example, where contemplated compositions are formulated as a solid product, suitable product forms include single dosage unit formulations such as capsules, tablets, and powders, while other solid formulations include snack bars, gummies, or other edible products onto which the composition is coated (e.g., onto cereal) or into which the composition is mixed or layered (e.g., into chewing gum). In another examples, where contemplated compositions are formulated as a liquid product, suitable product forms include flavored and/or carbonated beverages (e.g., tea, juice), functional beverages (e.g., sports or energy drinks) or infusions, or liquid dairy product (e.g., yoghourt, kefir).
Therefore, contemplated compositions may be provided in bulk, as part of an edible or drinkable product, and/or provided in single dosage units for consumption. Most typically, the daily dosage for contemplated compositions (excluding the carrier) is preferably at least 10 mg, or at least 100 mg, or at least 200 mg, or at least 300 mg, or at least 400 mg, or at least 500 mg, or at least 750 mg, or at least 1,000 mg, or at least 1,500 mg. For example, suitable dosages will be between 10-100 mg, or between 100-200 mg, or between 200-400 mg, or between 300-600 mg, or between 400-800 mg, or between 600-1,000 mg, or between 1,000-2,000 mg.
As will be readily appreciated, contemplated compositions may further be combined with one or more additional ingredients to impart further desirable functionalities, and suitable additional ingredients include vitamins (e.g., single vitamins, or vitamin blends such as multivitamin blends), dietary trace elements or minerals (e.g., individual elements or minerals, or mixtures of multiple elements or minerals in various forms), various specialty compounds and mixtures (e.g., compositions comprising nicotinamide riboside, prebiotics, human milk oligosaccharides), and/or one or more probiotic microorganisms (e.g., Lactobacillus spec., Bifidobacterium spec., Leukonostoc spec., Saccharomyces boulardii, etc.).
Of course, it should be recognized that the compositions according to the inventive subject matter may be administered not only to a human, but also to other non-human mammals, and especially livestock and companion animals (e.g., dogs, cats, horses). Administration will typically be between once daily and three times daily (and in some cases even more) over a period of at least two days, three days, five days, 1 week, 2-4 weeks, 1-3 months, and even longer. Most typically, administration can be performed for a period sufficient to provide at least symptomatic relief of a condition (e.g., pain and swelling associated with inflammation, low energy level, frequent infections, etc.), or prophylactically to avoid or help reduce severity of a health condition.

EXAMPLES

Representative Composition:

Unless indicated otherwise, all tests were performed with a defined mixture of selected polyphenol-containing products/extracts common to the Mediterranean diet. The polyphenol-containing products/extracts were obtained from source materials characterized by color as follows: Red group: apple extract, pomegranate extract, tomato powder, and beet root; Green group: olive extract, rosemary extract, green coffee bean extract, and kale; Orange/yellow group: onion extract, ginger extract, grapefruit extract, and carrot; and Purple/blue group: grape extract, blueberry extract, currant, and elderberry. Corn starch, silica, and sunflower lecithin were used as processing aids. Relative proportions are shown in Table 1 below.

TABLE 1

Ingredient	Wt %	Part	Solvent	Standardized to	State

Grape Extract	15.75	Whole	Ethanol	Min	50% total	Powdered
		fruit		Proanthocyanidins
Apple Extract	10.00	Whole	Ethanol/	70% Polyphenols	Powdered
		fruit	Water
Ginger Extract	10.00	Root	Ethanol/	5% Gingerol	Powdered
			Water
Onion Extract	10.00	Root	Ethanol/	30% Quercetin	Powdered
		bulb	Water
Pomegranate	10.00	Whole	Ethanol		40% Punicalagins	Powdered
Extract		fruit
Green Coffee	7.50	Seed	Ethanol/	50% Chlorogenic	Powdered
Bean Extract			Water	Acids
Rosemary	7.50	Leaves	Ethanol/	15% Rosmarinic	Powdered
Extract			Water	Acid
Olive Extract	6.38	Whole	Water	12%	Powdered
		fruit		Hydroxytyrosol
Maltodextrin	3.19	—/—	—/—	—/—	Powdered
Blueberry	2.50	Whole	Ethanol/	12% Anthocyanins	Powdered
Extract		fruit	Water
Grapefruit	2.50	Whole	Ethanol/	90% Naringin	Powdered
Extract		fruit	Water
Kale	2.50	Leaves	None	—/—	Powdered
Beet Root	2.45	Root	None	—/—	Powdered
Carrot	2.45	Root	None	—/—	Powdered
Black Currant	2.43	Whole	None	—/—	Powdered
		fruit
Elderberry	2.43	Whole	None	—/—	Powdered
		fruit
Tomato	1.85	Whole	None	—/—	Powdered
		fruit
Corn Starch	0.31	—/—	—/—	—/—	Powdered
Silica	0.25	—/—	—/—	—/—	Powdered
Sunflower	0.03	—/—	—/—	—/—	Powdered
Lecithin

Phytochemical HPLC/MS/MS analysis: An HPLC/MS compositional analysis of the exemplary composition above revealed the following ingredients and proportions where the columns in each of Tables 2-8 indicate the analyte ID (col.1), chemical entity (col.2), M-H (col.3), RT (col.4), peak intensity (col.5), and MS/MS fragments (col.6):

TABLE 1

Ingredient	Wt %	Part	Solvent	Standardized to	State

Grape Extract	15.75	Whole fruit	Ethanol	Min	50% total	Powdered
				Proanthocyanidins
Apple Extract	10.00	Whole fruit	Ethanol/Water	70% Polyphenols	Powdered
Ginger Extract	10.00	Root	Ethanol/	5% Gingerol	Powdered
			Water
Onion Extract	10.00	Root bulb	Ethanol/	30% Quercetin	Powdered
			Water
Pomegranate Extract	10.00	Whole fruit	Ethanol		40% Punicalagins	Powdered
Green Coffee Bean	7.50	Seed	Ethanol/Water	50% Chlorogenic	Powdered
Extract				Acids
Rosemary Extract	7.50	Leaves	Ethanol/	15% Rosmarinic Acid	Powdered
			Water
Olive Extract	6.38	Whole fruit	Water	12% Hydroxytyrosol	Powdered
Maltodextrin	3.19	—/—	—/—	—/—	Powdered
Blueberry Extract	2.50	Whole fruit	Ethanol/Water	12% Anthocyanins	Powdered
Grapefruit Extract	2.50	Whole fruit	Ethanol/Water	90% Naringin	Powdered
Kale	2.50	Leaves	None	—/—	Powdered
Beet Root	2.45	Root	None	—/—	Powdered
Carrot	2.45	Root	None	—/—	Powdered
Black Currant	2.43	Whole fruit	None	—/—	Powdered
Elderberry	2.43	Whole fruit	None	—/—	Powdered
Tomato	1.85	Whole fruit	None	—/—	Powdered
Corn Starch	0.31	—/—	—/—	—/—	Powdered
Silica	0.25	—/—	—/—	—/—	Powdered
Sunflower Lecithin	0.03	—/—	—/—	—/—	Powdered

TABLE 3

Organic acids

1	Citric acid	191.0186	2.9	5.25E+08	102, 111, 129, 173
2	Malic acid	133.0130	2.2	1.26E+08	71, 89, 115
3	Glucaric acid	209.0291	1.7	8.68E+08	133, 147, 191
4	Gluconic acid	195.0504	1.7	4.44E+08	99, 129, 159, 177
5	hydroxy jasmonic	387.1668	22.3	2.23E+08	59, 163, 207
	acid-O-glucoside
6	Azelaic acid	187.0968	28.3	1.56E+07	125, 169
5	Arabinonic acid	165.0395	1.7	1.96E+08	129, 147, 165

TABLE 4

Phenolics

7	Coumaric acid	163.0389	24.3	9.19E+06	119,147
8	Coumaric aicd-derv	295.0460	21.1	2.45E+06	119, 163
9	Quinic acid-I	191.0553	1.8	4.76E+08	173
10	Quinic acid-II	191.0553	20.1	2.40E+08	173
11	Caffeic acid	179.0341	21.4	1.54E+08	135
12	Caffeic acid-hexose-I	341.0869	16.6	5.02E+06	135, 161, 179
13	Caffeic acid-hexose-II	341.0869	18.5	1.28E+07	135, 161, 179
14	Caffeic acid-hexose-III	341.0869	19.6	1.02E+07	135, 179
15	Caffeic acid-hexose-IV	341.0869	20.0	1.61E+07	135, 161, 179
16	Ferulic acid	193.0498	25.6	4.32E+06	134.04,
					149.06,
					178.03
17	Ferulic acid-hexose-I	355.1028	21.7	1.97E+07	175
18	Ferulic acid-hexose-I	355.1028	22.5	5.63E+06
19	Ferulic acid-hexose-I	355.1028	23.3	6.47E+06	134, 149, 175
20	Ferulic acid-hexose-I	355.1028	24.9	3.24E+06	193
21	Ferulic acid-hexose-I	355.1028	25.9	4.32E+06	193
22	3,4-Dihydroxybenzoic	153.0188	10.2	1.23E+07	109
	acid
23	Gallic acid	169.0132	5.3	6.41E+07	125
24	Rosmaric acid	359.0772	28.8	1.18E+09	135, 131,
					179, 197
25	Rosmaric acid dimer	719.1618	28.8	5.48E+08	135, 161,
					179, 197
26	Salvianic acid A	197.0447	9.2	7.12E+07	123, 135,
					179, 180
27	Ethyl gallate	197.0449	24.3	1.83E+07	125, 151, 169
28	Ellagic acid	300.9994	25.8	1.20E+08	229, 257, 283
29	Ellagic acid glucoside	463.0529	22.9	5.69E+05	125, 169,
					300.99, 463
30	Galloyl-HHDP-	633.0737	22.7	6.10E+07	125, 169, 193,
	glucoside				275, 300.99
31	Digalloyl-HHDP-	785.0867	22.3	1.92E+06	125, 169,
	glucoside				275, 300.99
32	3-Galloylquinic acid	343.0673	5.9	1.70E+06	169, 191
	(Theogallin)
33	3,4-Dihydroxybenzoic	153.0183	10.2	1.23E+07	96, 109
34	3,4-Dihydroxybenzoic	153.0545	10.3	3.62E+08	109, 123,
					153.0182
35	2,4-Dihydroxybenzoic	153.0183	23.8	8.71E+06	109
36	2-Hydroxybenzoic acid	137.0232	15.0	1.10E+07	93

TABLE 5

Flavonoids

37	Naringenin	271.0604	34.4	2.96E+07	119, 151,
					227, 271
38	Naringenin-glucoside	433.1149	28.0	4.14E+06	151, 271
39	Naringenin-	579.1722	27.5	1.13E+09	151, 271,
	rhamnoglucoside				549
40	Naringenin-	1159.3540	27.5	5.66E+07	151, 271,
	rhamnoglucoside				459
	dimer
41	Myricetin	317.0503	29.0	1.09E+07	137, 151,
					178
42	Myricetin 3-rhamnoside	463.0894	25.8	5.61E+06	316, 317
43	Myricetin 3-glucoside I	479.0839	24.3	4.59E+06	316, 317
44	Myricetin 3-rhamnoside	479.0839	24.5	4.57E+06	316, 317
	II
45	Phloretin	273.0769	34.5	2.41E+08	167
46	Phloretin-glucoside	435.1298	29.0	2.85E+08	167, 273
47	Phloretin-arabinose-	567.1733	27.5	3.28E+08	167, 273
	glucoside
48	Phloretin-diglucoside	597.1831	26.8	7.70E+06	167, 273
49	Apigenin	269.0460	34.4	4.46E+07	151
50	Apigenin glucoside	431.0985	27.8	1.36E+07	268, 269
51	Apigenin rhamnoside	577.1575	27.4	8.96E+07	269
	glucoside
52	Apigenin rhamnoside	623.1635	27.4	7.88E+06	269, 577
	glucoside der
53	Apigenin rhamnoside	1155.3230	27.4	1.41E+06	269, 577
	glucoside dimer
54	Quercetin	301.0348	32.1	1.89E+09	151, 179,
					273, 301
55	Quercetin dimer	603.0795	32.1	3.94E+08	151, 178,
					301
56	Quercetin galactoside	463.0874	24.6	1.00E+07	301
57	Quercetin glucoside-1	463.0874	25.9	2.98E+07	151, 300,
					301
58	Quercetin glucoside-2	463.0874	26.1	7.16E+07	151, 300,
					301
59	Quercetin-diglucoside	625.1400	23.8	8.83E+05	151, 178,
					301, 463
60	Quercetin-rutinoside	609.1478	25.5	3.37E+07	300, 301
	or Rutin
61	Luteolin	285.0407	31.9	4.20E+07	133.03,
					199.04,
					217.05,
					241
62	Luteolin glucoside 1	447.0938	26.2	4.58E+06	284, 285
63	Luteolin glucoside 2	447.0938	24.6	1.50E+07	284, 285
64	Catechin-1	289.0704	20.2	5.97E+08	151, 179,
					205, 245
65	Catechin-2	289.0704	22.7	8.47E+08
66	Proanthocyanidin B1	577.1364	22.0	4.06E+08	125, 289,
					407
67	Proanthocyanidin B2	577.1364	18.9	2.14E+08	125, 289,
					407
68	Isorhamnetin	315.0503	35.4	2.66E+08	300, 315
69	Isorhamnetin glucoside	477.1030	26.7	1.11E+08	119, 151,
					299, 314,
					315
70	Isorhamnetin	639.1563	27.4	1.11E+06	151, 285,
	diglucoside-1				313, 315,
					476, 477,
					639
71	Isorhamnetin-	623.1939	26.1	6.63E+06	315
	rhamnosyl-glucoside
72	Kaempferol	285.0397	35.0	1.27E+08	125.02,
					244,
					257, 268
73	Kaempferol glucoside-1	447.0938	26.9	4.02E+06
74	Kaempferol glucoside-2	447.0938	27.4	1.98E+07
75	Kaempferol glucoside-3	447.0938	27.6	1.04E+07
76	Kaempferol glucoside-4	447.0938	27.9	4.47E+06
77	Kaempferol-3-	593.1521	25.6	1.68E+07	285
	O-rutinoside
78	Kampferol	755.2060	25.9	1.23E+06	285
	3-(6-glucosylglucoside)
	7-rhamnoside
79	wogonin	283.0616	41.2	1.42E+08	268, 283

TABLE 6

Anthocyanin

80	Cyanidin-3-O-glucoside	449.1073	21.1	1.67E+07	287
81	Cyanidin derv	463.0874	27.7	3.56E+05	287
82	Cyanidin derv	463.0874	29.1	1.97E+07	287
83	Cyanidin-3-O-rutinoside	595.1658	25.6	8.57E+05	287, 449
84	Cyanidin 3-sambubioside	581.1500	21.1	5.95E+06	287
85	Pelargonidin	271.0600	24.5	2.10E+06	271
86	Pelargonidin-glucoside	433.1131	27.9	7.85E+05	271
87	Pelargonidin-glucoside	639.1710	31.4	5.30E+05	175, 207,
	derv				271
88	Malvidin	331.0809	26.7	2.27E+06	316
89	Malvidin arabinoside	463.1237	23.0	8.98E+06	331
90	Malvidin-feruloyl-	639.1710	28.1	7.34E+05	331
	arabinoside
91	Delphinidin	303.0496	32.2	2.73E+07	303
92	Delphinidin 3-glucoside	465.1028	20.1	6.49E+06	303, 385
93	Delphinidin 3-rutinoside	611.1603	25.5	1.30E+06	303
94	Delphinidin-3-arabinoside-I	435.0922	20.7	2.06E+06	303
95	Petunidin	317.0655	35.5	2.77E+06	317
96	Petunidin 3-glucoside	479.1186	26.7	1.32E+07	317
97	Petunidin 3-rutinoside	625.1765	26.1	5.63E+05	317
98	Petunidin derv	463.0874	26.9	4.03E+05	317
99	Peonidin	301.0709	34.8	1.63E+06	286
100	Peonidin-glucoside-I	463.1240	26.9	2.77E+06	301
101	Peonidin-glucoside II	463.1240	28.3	6.98E+06	301
102	Peonidin-rutinoside	609.1814	27.6	1.52E+06	301
103	Peonidin-feruloyl-glucoside	639.1710	31.7	1.88E+06	177, 301

TABLE 7

Chlorogenic acids

104	3-Caffeoylquinic acid	353.0875	14.3	2.70E+08	135, 179, 191
105	5-Caffeoylquinic acid	353.0875	20.1	8.52E+08	179, 191
106	4-Caffeoylquinic acid	353.0875	21.1	5.09E+08	135, 173,
					179, 191
107	3-Caffeoylquinic	707.1830	14.3	1.87E+07	135, 179,
	acid dimer				191,353
108	5-Caffeoylquinic	707.1833	20.1	9.23E+08	191, 353
	acid dimer
109	4-Caffeoylquinic	707.1834	21.1	1.95E+08	135, 173,
	acid dimer				179, 191,
					353
110	3-Caffeoylshikimic acid	335.0770	24.2	3.17E+07	135, 161,
					173, 179
111	4-Caffeoylshikimic acid	335.0770	24.5	1.62E+08	135, 161
112	5-Caffeoylshikimic acid	335.0770	25.0	1.53E+08	161
113	3,4-Dicaffeoylshikimic	497.1091	29.5	0
	acid
114	3,5-Dicaffeoylshikimic	497.1091	32.0	9.10E+07	161, 179 335
	acid
115	4,5-Dicaffeoylshikimic	497.1091	32.7	6.97E+06	135, 161,
	acid				179, 335
116	3-Feruloylquinic acid	367.1032	20.7	1.13E+08	134, 193
117	5-Feruloylquinic acid	367.1032	23.6	3.76E+08	163, 191
118	4-Feruloylquinic acid	367.1035	23.8	1.66E+08	173, 193
119	3-Feruloylquinic	735.2144	20.7	7.14E+05
	acid dimer
120	5-Feruloylquinic	735.2144	23.6	1.03E+08	173, 191, 367
	acid dimer
121	4-Feruloylquinic	735.2144	23.8	4.17E+06	173, 193, 367
	acid dimer
122	3-Coumaroylquinic acid	337.0932	19.0	2.68E+07	119, 163
123	4-Coumaroylquinic acid	337.0933	22.6	9.95E+07	191
124	5-Coumaroylquinic acid	337.0933	23.0	4.09E+08	163, 173
125	3-Coumaroylquinic	675.1941	19.0	0
	acid dimer
126	4-Coumaroylquinic	675.1941	22.6	3.00E+06	191
	acid dimer
127	5-Coumaroylquinic	675.1941	23.0	1.53E+07	163, 173
	acid dimer
128	3,4-Dicaffeoyl	515.1189	27.1	2.63E+08	135, 161,
	quinic acid				173, 179,
					191,
					353
129	3,5-Dicaffeoyl	515.1190	27.6	1.79E+08	135, 179,
	quinic acid				191, 353
130	4,5-Dicaffeoyl	515.1195	28.3	3.37E+08	135, 173,
	quinic acid				179,
					191, 353
131	Valeroylquinic	275.1135	14.9	2.70E+05	101, 173, 181
	acid isomer-1
132	Valeroylquinic	275.1135	15.8	5.75E+05	101, 181, 191
	acid isomer-2
133	Valeroylquinic	275.1135	21.0	1.17E+06	101, 173
	acid isomer-3
134	Valeroylquinic	275.1135	21.4	2.26E+06	101, 173
	acid isomer-4
135	Valeroylquinic	275.1135	23.2	3.27E+05	93, 101, 173,
	acid isomer-5				181, 191
136	Valeroylquinic	275.1135	23.4	5.25E+05	101, 173
	acid isomer-6
137	Valeroylquinic	275.1135	30.4	5.22E+06	93, 101,
	acid isomer-7				173,
					181, 191
138	Valeroylquinic	275.1135	30.6	1.18E+06	101
	acid isomer-8
139	Valeroylquinic	275.1135	31.7	1.98E+06	101, 173
	acid isomer-9
140	Caffeoylvaleroylquinic	437.1451	29.7	3.55E+06	161, 173,
	acid isomer 1				179, 275
141	Caffeoylvaleroylquinic	437.1451	29.8	6.88E+06	161, 173,
	acid isomer 2				179, 275
142	Caffeoylvaleroylquinic	437.1451	30.4	2.46E+06	161, 173,
	acid isomer 3				179, 275
143	Caffeoylvaleroylquinic	437.1451	30.6	5.13E+06	173, 275
	acid isomer 4
144	Caffeoylvaleroylquinic	437.1451	31.7	6.96E+06	173, 275
	acid isomer 5
145	Quinic	481.2442	24.8	3.81E+07	161, 197, 301
	acid-glucoside-R*-1
146	Quinic	481.2442	25.3	1.22E+08	161, 197, 301
	acid-glucoside-R*-2
147	Valeroylquinic acid	565.3023	33.3	1.18E+07	301, 463, 481
	glucoside-R*-1
148	Valeroylquinic acid	727.3545	26.6	3.83E+06	161, 301,
	diglucoside-R*-1				323, 481, 643
149	Valeroylquinic acid	727.3545	30.4	1.89E+06	205, 361,
	diglucoside-R*-2				625, 643
150	Valeroylquinic acid	727.3545	30.8	3.05E+07	481, 523,
	diglucoside-R*-3				625, 643

TABLE 8

Betacyanin

151	Betanin	551.1517	15.6	2.19E+04
152	Isobetanin	551.1517	19.7	1.54E+04

TABLE 9

Amino acids, alkaloids & other compounds

153	Glycerophosphocholine	258.1098	1.7	6.17E+07	104
154	Adenosine	268.1040	4.1	4.68E+06	136
155	Phenylalanine	166.0861	6.8	1.41E+07	120
156	Tiyptophan	205.0973	14.5	6.28E+06	146, 188
157	Tyrosine	182.0812	3.7	9.16E+06	119, 123,
					136, 147, 165
158	Dopamine	154.0858	2.7	8.64E+05	113, 137
159	Trigonellin	138.0548	1.8	2.05E+08	94
160	Caffeine	195.0875	20.1	4.94E+07	138
161	Xanthine	151.0256	3.1	7.74E+05	109

Biological activity of the composition was tested for inhibition of various target entities that are central to various pathways associated with health and healthy ageing, and exemplary activity results are presented below. More specifically, the inventor tested the composition for inhibitory activity of human ARG-1, ARG-2, SIRT1, BACE1, Cathepsin S, CDK5, IDO1, IDO2, NAMPT, PCSK9, CD47, CD38, JAK1, JAK2, JAK3, CD39, and CD73, and for Keap/Nrf2 binding inhibition and ACE2-Spike binding inhibition.

ARG1 and ARG2:

In the following experiments, the inventor sought to determine whether the representative compositions had an effect on ARG1 and ARG2. Reagents used are shown in the Tables 9-10 below and tested as stated unless indicated otherwise (nor-NOHA is reference compound).

TABLE 10

	Form	Stock	Dissolving
Compound	Supplied	Conc.	Solvent	Test Range

HP Color Blend	powder		1% (w/v)	70% EtOH	0.0004, 0.002
lot#33890000X11020				and 0.01%
nor-NOHA*	powder	10 mM	DMSO	0.001, 0.01 and
				0.1 μM

TABLE 11

		Concentration
Enzyme	Lot#	(ng/well)	Substrate

ARG1	150825	100	Thioarginine (225 μM)
ARG2	160726-1	20	Thioarginine (225 μM)

Assay conditions: The assay was performed using human recombinant ARG1 or ARG2 as enzymes and thioarginine as substrate. The UV absorbance at 412 nm was correlated with the amount of reaction product of ARG1/ARG2. The test sample (HP Color Blend) was dissolved to 1% (w/v) with 70% (w/v) EtOH and filtered through 0.22 μm; nor-NOHA was dissolved in 100% (w/v) DMSO to 10 mM. 5 μl of 20× sample solutions were added to 90 μl of substrate, and reactions were started by the addition of 5 μl of 20× ARG1/ARG2 solutions. For the negative control (Blank), 5 μl of the assay buffer was added instead of the ARG1/ARG2. The resulting 100 μl reaction mixture contained the indicated amount of the samples, 225 μM thioarginine, the detection reagent, and 30 nM ARG1 or 5 nM ARG2 in 1× ARG Assay Buffer. All reactions were conducted at room temperature and incubated for 30 minutes, ensuring that all wells, containing either samples or controls, contained 0.7% (v/v) EtOH final concentration to discard any solvent effect. UV absorbance readings were done at times 0 and 30 minutes to get net values. Absorbance was measured using a Tecan Infinite M1000 microplate reader.
Data Analysis: Experiment was performed in duplicate at each concentration. The data were analyzed using the software GraphPad Prism. In the absence of the compound, the net absorbance (A_t) in each data set was defined as 100% activity. In the absence of ARG1/ARG2, the net absorbance (A_b) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=[(A−A_b)/(A_t−A_b)]×100, where A=the absorbance in the presence of the compound. The percent inhibition was calculated according to the following equation: % inhibition=100−% activity.
Results: Notably, significant inhibitory activity was found for both ARG-1 and ARG-2 across all tested concentrations as is shown in Tables 11-12 below. As can be readily seen from the results, inhibition relative to the reference inhibitor was significant and not specific towards one or the other of ARG-1 and ARG-2 as is also seen from Table 13. Results are also depicted in FIG. 1 and FIG. 2 for ARG-1 and ARG-2, respectively.

TABLE 12

	Net absorbance	Activity (%)	ARG-1

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	Inhibition (%)

No compound	0.76	0.75	101	99
HP Color Blend,	0.64	0.62	85	82	17
0.0004%
HP Color Blend,	0.56	0.55	74	74	26
0.002%
HP Color Blend,	0.51	0.50	68	66	33
0.01%
nor-NOHA,	0.74	0.77	99	102	0
0.001 μM
nor-NOHA, 0.01 μM	0.59	0.59	79	78	22
nor-NOHA, 0.1 μM	0.23	0.24	31	31	69
Blank	0.00	0.00

TABLE 13

	Net absorbance	Activity (%)	ARG-2

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	Inhibition (%)

No compound	1.01	0.93	104	96
HP Color Blend,	0.72	0.72	74	74	26
0.0004%
HP Color Blend,	0.59	0.64	60	66	37
0.002%
HP Color Blend,	0.63	0.59	65	60	38
0.01%
nor-NOHA,	0.78	0.82	81	84	17
0.001 μM
nor-NOHA, 0.01 μM	0.56	0.60	57	62	41
nor-NOHA, 0.1 μM	0.19	0.19	19	19	81
Blank	0.00	0.00	0	0

TABLE 14

	Inhibition (%)

Condition	ARG1	ARG2

HP Color Blend, 0.0004%	17	26
HP Color Blend, 0.002%	26	37
HP Color Blend, 0.01%	33	38
nor-NOHA, 0.001 μM	0	17
nor-NOHA, 0.01 μM	22	41
nor-NOHA, 0.1 μM	69	81

SIRT1:

In the following experiments, the inventor sought to determine whether the representative compositions had an effect on SIRT1. Reagents used are shown in Tables 14-15 below and tested as stated unless indicated otherwise (Suramin was used as reference compound).

TABLE 15

	Compound	Stock	Dissolving	Test Range
Compound I.D.	Supplied	Concentration	Solvent	Dilution	Intermediate

HP Color Blend	Solid	1% (w/v)	70%	0.0004%,	10% DMSO in
lot#33890000X11020			EtOH	0.002%, 0.01%	HDAC Assay
					Buffer
Suramin*	Solid	10 mM	DMSO	0.01 μM,	10% DMSO in
				0.1 μM, 1 μM	HDAC Assay
					Buffer

TABLE 16

	Catalog	Enzyme	Enzyme Used (ng)/
Assay	#	Lot #	Reaction	Substrate

SIRT1	50012	190710	550	10 μM HDAC
				Substrate
1

Assay Conditions: The sample was dissolved in 70% EtOH. The serial dilution of the compound was first performed in 70% EtOH with the highest concentration at 1%. Each intermediate compound dilution (in 70% EtOH) will then get directly diluted 10× fold into assay buffer for an intermediate dilution of 7% EtOH in HDAC assay buffer and 5 μl of the dilution was added to a 50 μl reaction so that the final concentration of EtOH is 0.7% in all of reactions.
The enzymatic reactions for the SIRT1 enzyme were conducted in duplicate at 37° C. for 30 minutes in a 50 μl mixture containing SIRT assay buffer, 5 μg BSA, an HDAC substrate (see 2.3.1), a SIRT enzyme, and a test compound. After enzymatic reactions, 50 μl of 2× SIRT Developer was added to each well for the SIRT enzymes and the plate was incubated at room temperature for an additional 15 minutes. Fluorescence intensity was measured at an excitation of 360 nm and an emission of 460 nm using a Tecan Infinite M1000 microplate reader.
Data Analysis: SIRT activity assays were performed in duplicates at each concentration. The fluorescent intensity data were analyzed using the computer software, Graphpad Prism. In the absence of the compound, the fluorescent intensity (Ft) in each data set was defined as 100% activity. In the absence of SIRT, the fluorescent intensity (Fb) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(F−F_b)/(F_t−F_b), where F=the fluorescent intensity in the presence of the compound.
Results: The inhibitory results are shown in the table below. The percent inhibition of the compounds against SIRT1 are summarized. The reference compound and tested composition and HP Color Blend precipitated at the 0.01% intermediate dilution step. FIG. 3 depicts the results in graphic form, and Table 16 provides results in numerical format. As can be readily appreciated, the tested composition had noticeable SIRT1 inhibitory activity.

TABLE 17

	SIRT1 Activity		%
	(Fluorescence count)	% Activity	Inhi-

Compound I.D.	Repeat 1	Repeat 2	Repeat 1	Repeat 2	bition

No Compound	2394	2429	99	101	0
HP Color Blend,	2407	2382	100	99	1
0.0004%
HP Color Blend,	2199	2200	91	91	9
0.002%
HP Color Blend,	1821	1891	74	77	24
0.01%
Suramin, 0.01 RM	2415	2415	100	100	0
Suramin, 0.1 RM	2074	2082	85	86	14
Suramin, 1 RM	710	702	26	26	74
Background	109	110

Keap/Nrf2:

In a further set of experiments, the inventor investigated whether or not the tested composition was able to interfere with Keap1-Nrf2 binding, and exemplary results are provided below. Reagents used are shown in Table 17 below and tested as stated unless indicated otherwise (reference compound was LDEETGEFL-OH).

TABLE 18

	Com-	Stock	Dis-	Test	Inter-
	pound	Concen-	solving	Range	mediate
Compound I.D.	Supplied	tration	Solvent	(%)	Dilution

HP Color Blend	Powder		1%	70%	0.0004,	7%
lot#33890000X11020			Ethanol	0.002,	Ethanol
				0.01
Reference	Powder		10 mM	DMSO	0.1, 1, 10,	10%
				100	DMSO

Assay Conditions: The test compound is diluted in 7% ethanol, and 5 μl of the dilution was added to a 50 μl reaction so that the final concentration of ethanol is 0.7%. The reference compound is diluted in 10% DMSO, and 5 μl of the dilution was added to a 50 μl reaction so that the final concentration of DMSO is 1%. The binding reactions were conducted at room temperature for 30 minutes in a 50μl mixture containing 10 mM HEPES, pH7.4, 50 mM EDTA, 150 mM NaCl, 0.05% Tween20, 0.01% BSA, 100 nM Keap1, 5 nM fluorescence probe and the test compound. Fluorescence intensity was measured at an excitation of 475 nm and an emission of 520 nm using a Tecan Infinite M1000 microplate reader.
Data Analysis: All of binding assays were performed in 96-well plates in duplicate. Fluorescence intensity is converted to fluorescence anisotropy using the Tecan Magellan6 software. The fluorescence anisotropy data were analyzed using the computer software, Graphpad Prism. The fluorescence anisotropy (F_At) in the sample with KeapI and the probe in each data set was defined as 100% activity. The fluorescence anisotropy (F_Ab) in the sample with a compound but without KeapI in each data set was defined as 0% activity. The percent binding efficacy in the presence of the competitor compound was calculated according to the following equation
$% Activity = \frac{(F_{A} - F_{Ab})}{(F_{At} - F_{Ab})} X 100 %$
where F_A=the fluorescence anisotropy in the presence of the compound. The values of % binding were then plotted in a bar graph as shown in FIG. 4, and numerical results are shown in Table 18 below.

TABLE 19

	Background	Binding Activity
	Fluorescent	Fluorescent
	Polarization (mA)	Polarization (mA)	Percentage Activity	Percentage

Compound	Repeat1	Repeat2	Repeat2	Repeat2	Repeat2	Repeat2	Inhibition

No Compound	18	14	38	40	96	104	0
0.0004%	21	22	43	43	92	93	7
0.002%	38	41	48	50	39	48	56
0.01%	78	76	81	81	16	17	83

As will be readily recognized from the results in the Table above and FIG. 4, the tested composition had appreciable inhibitory activity against Keap1-Nfr2 binding.
ACE2-Spike:
In this series of experiments, the inventor investigated whether contemplated compositions and fractions thereof could reduce binding of SARS-CoV2 spike protein to ACE2, and if vitamins would also have any effect.
Reagents used are shown in Tables 19-21 below and tested as stated unless indicated otherwise (* denotes reference compounds). In this series of experiments, the representative composition as described above was compared against individual color as indicated below and further against a multivitamin formulation as also indicated below. Table 19 denotes the representative composition, Table 20 denotes the color subfractions of the representative composition in which DC-5=Yellow Blend, DC-9=Purple Blend, DC-21-Green Blend, DC-13=Red Blend. Here, the red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, Elderberry as also noted above. Table 21 denotes the multivitamin blend, and Table 22 denotes the ACE2/Spike reagents used.

TABLE 20

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.0008, 0.004,
lot#33890000X11020				0.02 and 0.1%
Anti-ACE2*	Powder	1.33 μM	PBS	0.27 μM
Spike S1*	Solution	5.6 μM	PBS	0.01, 0.1
				and 1 μM

TABLE 21

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DC-5	Solid	1% (w/v)	70% (v/v) EtOH	0.0008, 0.004,
				0.02 and 0.1%
DC-9	Solid	1% (w/v)	70% (v/v) EtOH	0.0008, 0.004,
				0.02 and 0.1%
DC-13	Solid	1% (w/v)	70% (v/v) EtOH	0.0008, 0.004,
				0.02 and 0.1%
DC-21	Solid	1% (w/v)	70% (v/v) EtOH	0.0008, 0.004,
				0.02 and 0.1%
Anti-ACE2*	Solid	13.3 μM	PBS	0.27 μM
Spike S1*	Solution	5.6 μM	PBS	0.01, 0.1
				and 1 μM

TABLE 22

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DC-TIV-1.0 (Adult	Powder		1% (w/v)	70% (v/v)	0.0008, 0.004,
Centrum Vitamin)			EtOH	0.02and 0.1%
Anti-ACE2*	Powder	13.3 μM	PBS	0.27 μM
Spike S1*	Solution	5.6 μM	PBS	0.01, 0.1 and
				1 μM

TABLE 23

Component	Catalog #	Lot #	Concentration

ACE2-His	11003	131001	50 ng/well
Spike 51-Biotin	100679	200326	40 nM

Assay Conditions: Nickel plate was coated at room temperature for 1 hour with 50 μl of 1 μg/ml of ACE2-His and washed and blocked before starting the reaction. 10 μl of compound solutions were incubated with 20 μl of 1× Immune Buffer in ACE2-His-coated assay wells during 30 minutes before starting the reaction by the addition of 20 μl of 5 μg/ml Spike S1-Spike. Controls with the same concentration of solvent (EtOH) were included in the study. Reaction was progressed for 1 hour at room temperature. Then, wells were washed three times with 1× Immune Buffer and blocked with Blocking Buffer 2 for 10 minutes. 100 μl of Streptavidin-HRP was added to all wells and incubated for 1 hour. Lastly, plate was emptied, washed three times and blocked before the addition of 100 μl of freshly prepared HRP chemiluminescent substrates to every well. Immediately, the luminescence of the samples was measured in a BioTek Synergy 2 microplate reader.
Data Analysis: The luminescence data were analyzed and compared. In the absence of the compound, the intensity (C_e) in each data set was defined as 100% activity. In the absence of enzyme, the intensity (C₀) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C₀)/(C_e−C₀), where C=the luminescence in the presence of the compound.
Results for the representative composition are shown in Table 23 and FIG. 5, while results for the multivitamin formulation are shown in Table 24 and FIG. 6. Results for the colored blends are shown in Table 25 and FIG. 7. As can be readily seen from the data in Tables 23-25 and FIGS. 5-7, the representative composition showed synergistic inhibition of ACE2-Spike S1 binding, particularly at lower concentrations, whereas the multivitamin formulation had no significant inhibitory effect. Moreover, selected DC-13 and DC9 also had some inhibitory effect per se.

TABLE 24

	Luminescence	Activity (%)	Inh.

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	(%)

No compound (0% EtOH)	73041	72023	101	99	0
#33890000X11020, 0.0008%	69278	67571	96	93	6
(0.06% EtOH)
#33890000X11020, 0.004%	61074	58147	84	80	18
(0.3% EtOH)
#33890000X11020, 0.02%	39077	40075	54	55	45
(1.5% EtOH)
#33890000X11020, 0.1%	4531	4376	6	6	94
(7% EtOH)
0.06% EtOH control	69515	66778	96	92	6
0.3% EtOH control	73813	68546	102	95	2
1.5% EtOH control	65414	71374	90	98	6
7% EtOH control	69262	72935	95	101	2
Anti-ACE2, 0.27 μM	44726	43081	62	59	40
Spike-Fc, 0.01 μM	65506	67497	90	93	8
Spike-Fc, 0.1 μM	44243	48002	61	66	36
Spike-Fc, 1 μM	11079	13329	15	18	83
Blank	53	59	0	0	100

TABLE 25

	Luminescence	Activity (%)	Inh.

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	(%)

No compound (0% EtOH)	69783	68734	101	99	0
DC-TIV-1.0, 0.0008%	68843	68470	99	99	1
(0.06% EtOH)
DC-TIV-1.0, 0.004%	67092	67275	97	97	3
(0.3% EtOH)
DC-TIV-1.0, 0.02%	68611	67415	99	97	2
(1.5% EtOH)
DC-TIV-1.0, 0.1% (7% EtOH)	65686	62732	95	91	7
0.06% EtOH control	67102	65413	97	94	4
0.3% EtOH control	67558	66286	98	96	3
1.5% EtOH control	68907	68688	99	99	1
7% EtOH control	68106	69743	98	101	0
Anti-ACE2, 0.27 μM	39545	38080	57	55	44
Spike-Fc, 0.01 μM	63863	63541	92	92	8
Spike-Fc, 0.1 μM	38990	44396	56	64	40
Spike-Fc, 1 μM	9850	10519	14	15	85
Blank	78	47	0	0	100

TABLE 26

	Luminescence	Activity (%)

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	Inh. (%)

No compound	78752	77962	101	99
(EtOH, 0%)
DC-5, 0.0008%	88634	87345	113	111	0
DC-5,0.004%	88490	87713	113	112	0
DC-5,0.02%	86311	82378	110	105	0
DC-5,0.1%	46242	32236	59	41	50
DC-9,0.0008%	88766	87246	113	111	0
DC-9, 0.004%	82412	80733	105	103	0
DC-9, 0.02%	65595	69238	84	88	14
DC-9,0.1%	3681	5643	5	7	94
DC-13, 0.0008%	83749	83790	107	107	0
DC-13, 0.004%	78717	76657	100	98	1
DC-13,0.02%	47967	53241	61	68	35
DC-13,0.1%	608	211	1	0	100
DC-21,0.0008%	84059	81164	107	104	0
DC-21, 0.004%	83790	81432	107	104	0
DC-21, 0.02%	78473	78360	100	100	0
DC-21,0.1%	66249	64005	85	82	17
EtOH, 0.06%	78473	82268	100	105	0
EtOH, 0.3%	80826	81863	103	104	0
EtOH, 1.5%	78508	78870	100	101	0
EtOH, 7%	71844	68213	92	87	11
Anti-ACE2, 0.27 μM	34327	36753	44	47	55
Spike-Fc, 0.01 μM	72198	74621	92	95	6
Spike-Fc, 0.1 μM	49403	50633	63	65	36
Spike-Fc, 1 μM	13755	11790	17	15	84
Blank	63	53

BACE1:

In a further set of experiments, the inventor sought to determine whether the representative compositions and fractions thereof as well as a multivitamin mix had an effect on the activity of recombinant human BACE1 using an in vitro fluorescence assay.
Reagents used are shown in Tables 26-28 below and tested as stated unless indicated otherwise (*Verubecestat was used as reference compound). Table 26 shows the representative composition, while Table 27 shows the colored fractions and multivitamin mix. Here, DC-5=Yellow Blend, DC-9=Purple Blend, DC-21-Green Blend, DC-13=Red Blend DCH-TIV 1.0 (Adult Centrum Multivitamins). The red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, and Elderberry as listed above.

TABLE 27

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.0004, 0.002
lot#33890000X11020				and 0.01%
Verubecestat*	Powder	10 mM	DMSO	0.01, 0.1 and
				1 μM

TABLE 28

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DC-5	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
				and 0.01%
DC-9	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
				and 0.01%
DC-13	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
				and 0.01%
DC-21	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
				and 0.01%
DCH-TIV 1.0	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
				and 0.01%
Verubecestat*	Powder	10 mM	100%	0.01, 0.1 and
			DMSO	1 μM

TABLE 29

Assay	Catalog #	Protein lot #	[E] (ng/well)	Substrate

BACE1	71657	170307	500	7.5 μM BACE1
				FRET peptide
				substrate

Assay Conditions: 80 μl of BACE1 was incubated with 10 μl of samples and reference compound for 10 minutes. Then, reaction was started by the addition of 10 μl of BACE1 FRET peptide substrate and product kinetics were measured for 1 hour in an Infinite M1000 microplate reader (Tecan). In Blank control wells, 80 μl of assay buffer was added instead of enzyme, all wells contained 0.7% (v/v) EtOH final assay concentration.
Data Analysis: All conditions were performed in duplicates at each concentration. The fluorescent intensity data was analyzed using Prism (GraphPad). In the absence of the compound (No compound control), the fluorescent intensity (Ft) in each data set was defined as 100% activity. In the absence of the enzyme (Blank control), the fluorescent intensity (Fb) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % Activity=(F−Fb)/(Ft−Fb), where F=the fluorescent intensity in the presence of the compound. Fluorescence at initial time was subtracted to obtain net signal, measured in relative fluorescence units (RLU).
As can be seen form the data in Table 29 and FIG. 8, the representative composition had substantial inhibitory activity against BACE1 as tested. Remarkably, as can be seen from the data in Table 30 and FIG. 9, the colored fractions also provided significant inhibition of BACE1. Moreover, it is also evident from these data that the BACE1 inhibition is synergistic, while the tested multivitamin had substantially no inhibitory effect.

TABLE 30

	Fluorescence
	(net RFU)	Activity (%)	Inh.

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	(%)

No compound	419	403	102	98	0
#33890000X11020, 0.0004%	166	143	39	34	63
#33890000X11020, 0.002%	69	66	15	15	85
#33890000X11020, 0.01%	47	61	10	13	88
Verubecestat, 0.01 μM	381	386	93	94	7
Verubecestat, 0.1 μM	168	158	40	37	61
Verubecestat, 1 μM	16	20	2	3	97
Blank	8	6

TABLE 31

	Fluorescence
	(net RFU)	Activity (%)

Condition	Rep. 1	Rep. 2	Rep. 1	Rep. 2	Inh. (%)

No compound	779	795	99	101
DC-5, 0.0004%	651	620	83	79	19
DC-5, 0.002%	671	636	85	81	17
DC-5, 0.01%	670	622	85	79	18
DC-9, 0.0004%	332	286	42	36	61
DC-9, 0.002%	138	142	17	18	83
DC-9, 0.01%	231	146	29	18	76
DC-13, 0.0004%	241	274	30	34	68
DC-13, 0.002%	184	176	23	22	78
DC-13, 0.01%	158	198	20	25	78
DC-21, 0.0004%	739	763	94	97	5
DC-21, 0.002%	592	613	75	78	24
DC-21,0.01%	436	375	55	47	49
DCH-TIV 1.0, 0.0004%	797	784	101	100	0
DCH-TIV 1.0, 0.002%	767	774	97	98	2
DCH-TIV 1.0, 0.01%	765	721	97	92	6
Verubecestat, 0.01 μM	821	793	104	101	0
Verubecestat, 0.1 μM	684	676	87	86	14
Verubecestat, 1 μM	27	31	3	3	97
Blank	8	1

Cathepsin S:

In yet another set of experiments, the inventor sought to determine whether the representative compositions and fractions thereof as well as a multivitamin mix had an effect on the activity of recombinant human Cathepsin S using an in vitro enzymatic assay.
Reagents used are shown in Tables 31-33 below and tested as stated unless indicated otherwise (*E-64 was used as reference compound). Here, the designations and ingredients of D5, D9, D13, and D21 are as noted above.

TABLE 32

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.004, 0.02
lot#33890000X11020				and 0.1%
E-64	Solution
	1 mM	DMSO				1, 10 and 100 nM

TABLE 33

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

D5	Powder
	1%	70% EtOH	0.0004, 0.002 and 0.01%
		(w/v)
D9	Powder		1%	70% EtOH	0.0004, 0.002 and 0.01%
		(w/v)
D13	Powder		1%	70% EtOH	0.0004, 0.002 and 0.01%
		(w/v)
D21	Powder		1%	70% EtOH	0.0004, 0.002 and 0.01%
		(w/v)
E-64	Solution		1 mM	DMSO		1, 10 and 100 nM

TABLE 34

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-	Powder	1% (w/v)	70% EtOH	0.004, 0.02
TIV-0.5				and 0.1%
DCH-	Powder	1% (w/v)	70% EtOH	0.004, 0.02
TIV-1.0				and 0.1%
(Adult
Centrum
Multi-
vitamin)
E-64*	Solution	1 mM	DMSO	0.001, 0.01
				and 0.1 μM

Assay Conditions: Cathepsin S was activated by diluting the concentrated storage stock into the acidic assay buffer for 30 minutes at room temperature. Then, 5 μl of the sample or reference inhibitor was added to 20 μl of enzyme solution and pre-incubated for 30 minutes. The enzymatic reactions were started by the addition of 25 μl of the fluorogenic substrate, for a total reaction volume of 50 μl. Reaction time was 60 minutes, and then fluorescence intensity at an excitation of 360 nm and an emission of 460 nm was read using a Tecan Infinite M1000 microplate reader.
Data Analysis: Enzyme activity assays were performed in duplicates at each concentration. The fluorescence intensity data were analyzed and compared. In the absence of the compound, the intensity in each data set was defined as 100% (Ce) activity. In the absence of enzyme, the intensity in each data set was defined as 0% (C0) activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C0)/(Ce−C0), where C=the intensity in the presence of the compound (all percent activities below zero were shown zero in the table). Compound fluorescence was removed by subtracting fluorescence at reaction time=0.
The results for the above tests are shown in Tables 34-36, with Table 34 and FIG. 10 showing results for the representative composition, Table 35 and FIG. 11 showing results for the various colored fractions, and Table 36 and FIG. 12 showing results for the multivitamin mixture (DCH-TIC-0.5 is representative composition; DCH-TIC-1.0 is Centrum multivitamin mix). As can be readily seen form the results, the representative composition as well as colored blends D9 and D13 had significant inhibition of Cathepsin S, whereas the multivitamin mix had comparably no substantial effect.

TABLE 35

	Net Signal

(Fluorescence counts)

Activity (%)

Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	10117	10153	100	100
#33890000X11020, 0.0004%	2866	2790	28	27	72
#33890000X11020, 0.002%	543	520	5	5	95
#33890000X11020, 0.01%	202	218	2	2	98
E-64. 1 nM	9894	9604	98	95	4
E-64, 10 nM	5170	5444	51	54	48
E-64, 100 nM	958	960	9	9	91
Blank	10	7

TABLE 36

	Net Signal	Inhibi-

(Fluorescence counts)

Activity (%)

tion

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	9022	8921	101	99
D5, 0.0004%	7844	8711	87	97	8
D5, 0.002%	7888	7297	88	81	15
D5, 0.01%	7040	7451	78	83	19
D9, 0.0004%	1377	1759	15	20	83
D9, 0.002%	447	446	5	5	95
D9, 0.01%	222	238	2	3	97
D13, 0.0004%	883	914	10	10	90
D13, 0.002%	271	314	3	3	97
D13, 0.01%	128	131	1	1	99
D21, 0.0004%	8732	8446	97	94	4
D21, 0.002%	7211	6789	80	76	22
D21, 0.01%	956	736	11	8	91
E-64, 1 nM	8864	8931	99	100	1
E-64, 10 nM	5727	5326	64	59	38
E-64, 100 nM	1009	1033	11	12	89
Blank	0	0

TABLE 37

	Net Signal	Inhibi-

(Fluorescence counts)

Activity (%)

tion

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	60213	59191	101	99	0
DCH-TIC-0.5,	37364	38895	63	65	36
0.0004%
DCH-TIC-0.5,	13816	12919	23	22	78
0.002%
DCH-TIC-0.5,	1258	1445	2	2	98
0.01%
DCH-TIC-1.0,	61364	58864	103	99	0
0.0004%
DCH-TIC-1.0,	61972	62138	104	104	0
0.002%
DCH-TIC-1.0,	41515	43925	69	74	28
0.1%
E-64, 0.001 μM	52418	49644	88	83	15
E-64, 0.01 μM	21262	19656	36	33	66
E-64, 0.1 μM	3091	3478	5	6	95
Blank	37	114

CDK5:

In a further set of experiments, the inventor sought to determine whether the representative compositions and fractions thereof as well as a multivitamin mix had an effect on the enzymatic activities of recombinant human CDK5/p25 using an in vitro enzymatic assay.
Reagents used are shown in Tables 37-39 below and tested as stated unless indicated otherwise (Dinaciclib was used as reference compound). Here, the designations and ingredients of D5, D9, D13, and D21 are as noted above.

TABLE 38

	Compound	Stock	Dissolving		Intermediate
Compound I.D.	Supplied	Concentration	Solvent	Test Range	Dilution

HP Colors	Solution		1%	70% Ethanol	0.01%, 0.002%,	Water
X11020				0.0004%
Dinaciclib	Solution
	10 mM	DMSO		0. 002 μM,	10%
				0.02 μM, 0.2 μM	DMSO (aq)

TABLE 39

	Compound	Stock	Dissolving		Intermediate
Compound I.D.	Supplied	Concentration	Solvent	Test Range	Dilution

DC-5	Solution	1%	70% Ethanol	0.01%,	Water
				0.002%,
				0.0004%
DC-9	Solution	1%	70% Ethanol	0.01%,	Water
				0.002%,
				0.0004%
DC-13	Solution	1%	70% Ethanol	0.01%,	Water
				0.002%,
				0.0004%
DC-21	Solution	1%	70% Ethanol	0.01%,	Water
				0.002%,
				0.0004%
DCH-TIV 1.0	Solution	1%	70% Ethanol	0.01%,	Water
(Adult Centrum				0.002%,
Multivitamin)				0.0004%
Dinaciclib	Solution
	10 mM	DMSO	0.002 μM,	10% DMSO
				0.02 μM,	(aq)
				0.2 μM,

TABLE 40

	Catalog #	Enzyme Used/
Assay	(Lot #)	Reaction (ng)	Substrate

CDK5/p25	40105	10	0.1 mg/ml CDK
	(130618-2)		Substrate Peptide 1
			10 μM ATP

Assay Conditions: The assay was performed using Kinase-Glo Plus luminescence kinase assay kit (Promega). It measures kinase activity by quantitating the amount of ATP remaining in solution following a kinase reaction. The luminescent signal from the assay is correlated with the amount of ATP present and is inversely correlated with the amount of kinase activity. The reference compound was diluted to 10% DMSO and 5 μl of the dilution was added to a 50 μl reaction so that the final concentration of DMSO is 1% in all of reactions. The test compound was diluted in water and 5 μl of the dilution was added to a 50 μl reaction. All of the enzymatic reactions were conducted at 30° C. for 45 minutes. The 50 μl reaction mixture contains 40 mM Tris, pH 7.4, 10 mM MgCl2, 0.1 mg/ml BSA, 1 mM DTT, 10 μM ATP, Kinase substrate and the enzyme. After the enzymatic reaction, 50 μl of Kinase-Glo Plus Luminescence kinase assay solution (Promega) was added to each reaction and incubated for 15 minutes, on the plate, at room temperature. Luminescence signal was measured using a BioTek Synergy 2 microplate reader.
Data Analysis: Kinase activity assays were performed in duplicate at each concentration. The luminescence data were analyzed using the computer software, Graphpad Prism. The difference between luminescence intensities in the absence of Kinase (Lut) and in the presence of Kinase (Luc) was defined as 100% activity (Lut−Luc). Using luminescence signal (Lu) in the presence of the compound, % activity was calculated as: % activity={(Lut−Lu)/(Lut−Luc)}×100%, where Lu=the luminescence intensity in the presence of the compound (all percent activities below zero were shown zero in the table).
The results for the above tests are shown in Tables 40-42, with Table 40 and FIG. 13 showing results for the representative composition, Table 41 and FIG. 14 showing results for the various colored fractions and the multivitamin mixture (here denoted as DCH-TIV 1.0). As can be readily taken from the data presented, the representative composition and the fractions thereof had significant inhibitory effect on CDK5, whereas the multivitamin mix had substantially no appreciable inhibitory effect as compared to the representative composition. Moreover, and at least at medium and low concentrations, the CDK5 inhibition had an at least moderate synergistic effect in the representative composition.

TABLE 41

	Kinase Activity

Luminescence

% Activity

%

Compounds	Repeat1	Repeat2	Repeat1	Repeat2	Inhibition

No Compound	21499	21364	100	100
HP Colors X11020,	26178	26281	73	73	27
0.0004%
HP Colors X11020,	33942	33967	29	29	71
0.002%
HP Colors X11020,	38055	38177	6	5	94
0.01%
Dinaciclib, 0.002 μM	28384	28205	61	62	39
Dinaciclib, 0.02 μM	36896	36850	12	13	87
Dinaciclib, 0.2 μM	37268	37490	10	9	90
Background	38991	39210

TABLE 42

	Kinase Activity

Luminescence

% Activity

Compounds	Repeat1	Repeat2	Repeat1	Repeat2	% Inhibition

No Compound	20187	20295	100	100
0.0004% DC-5	23997	23914	81	82	19
0.002% DC-5	35968	35411	22	24	77
0.01% DC-5	40138	40324	1	0	100
0.0004% DC-9	22139	22596	91	88	11
0.002% DC-9	27328	27981	65	61	37
0.01% DC-9	36274	36077	20	21	79
0.0004% DC-13	25412	25396	74	74	26
0.002% DC-13	28786	28580	57	58	42
0.01% DC-13	33831	33995	32	31	68
0.0004% DC-21	20168	20357	100	99	0
0.002% DC-21	22651	22070	88	91	11
0.01% DC-21	30462	30543	49	49	51
0.0004%	20553	20979	98	96	3
DCH-TIV 1.0
0.002%	20929	21076	97	96	4
DCH-TIV 1.0
0.01% DCH-	21472	21736	94	93	7
TIV 1.0
Dinaciclib,	29310	29124	55	56	45
0.002 μM
Dinaciclib,	38603	39041	8	6	93
0.02 μM
Dinaciclib,	40248	39973	0	2	99
0.2 μM
Background	40474	40099

IDO1/IDO2:

In a further set of experiments, the inventor sought to determine whether the representative compositions had an effect on the enzymatic activities of recombinant human IDO1 and/or IDO2 using an UV absorbance assay.
Reagents used are shown in Tables 42-43 below and tested as stated unless indicated otherwise.

TABLE 43

		Stock
	Compound	Concentration		Test	Intermediate
Compound I.D.	Supplied	(mM)	Solvent	concentration	Dilution

DailyColors Blend	Liquid		1% (w/v)	70%	0.01%, 0.002%	20% DMSO
Lot # 33890000X11020			EtOH	0.0004%
INCB024360*	Liquid	0.05	DMSO	0.01, 0.1, 1,	20% DMSO
				10 μM

TABLE 44

			Enzyme
	Catalog	Lot	concentration
Assay	#	#	(nM)	Substrate

IDO1	71182	180305-B	40	L-Tryptophan (4 mM)
IDO2	71194	160519-C	400	L-Tryptophan (4 mM)

Assay Conditions: The assay was performed measuring UV absorbance using recombinant IDO and L-Tryptophan as substrate. The UV absorbance at 321 nm is correlated with the amount of N-formylkynurenine, reaction product of IDO. The compounds (see 2.2) were diluted in 20% DMSO and 10 μl of the dilution was added to a 200 μl reaction so that the final concentration of DMSO was 1% in all reactions. All of the reactions were conducted at room temperature. The 200 μl reaction mixture in IDO Assay Buffer contained 400 nM IDO1 or IDO2, the indicated amount of the inhibitor, tryptophan, and the coupled reaction components. The reaction mixture incubated was for 180 min prior to reading the UV absorbance. For the negative control (blank), 10 μl of the assay buffer was added instead of the IDO enzyme. Absorbance was measured using a Tecan Infinite M1000 plate reader.
Data Analysis: The experiments were performed in duplicate at each concentration. The data were analyzed using the computer software GraphPad Prism. In the absence of the compound, the absorbance signal (At) in each data set was defined as 100% activity. In the absence of the IDO, the absorbance signal (Ab) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=[(A−Ab)/(At−Ab)]×100, where A=the absorbance signal in the presence of the compound. The percent inhibition was calculated according to the following equation: % inhibition=100−% activity.
The results for the above tests are shown in Tables 44-45, with Table 44 and FIG. 15 showing results for the representative composition on inhibition of IDO1, and Table 45 and FIG. 16 showing results for the representative composition on inhibition of IDO2. As can be readily taken from the data presented, the representative composition had inhibitory effect on IDO1 at higher concentrations and significant inhibitory effect on IDO2 at higher and moderate concentrations.

TABLE 45

	Absorbance (net)	Activity (%)	%

Compound I.D.	Repeat 1	Repeat 2	Repeat 1	Repeat 2	Inhibition

No Compound	0.82	0.76	104	96	0
DailyColors Blend,	1.13	1.15	145	147	0
0.0004%
DailyColors Blend,	0.87	0.89	111	113	0
0.002%
DailyColors Blend,	0.57	0.56	71	70	30
0.01%
INCB024360,	0.73	0.74	92	93	7
0.1 μM
INCB024360,	0.29	0.30	34	36	65
1 μM
INCB024360,	0.07	0.09	5	8	93
10 μM
Blank	0.03	0.03

TABLE 46

	Absorbance (net)	Activity (%)	%

Compound I.D.	Repeat 1	Repeat 2	Repeat 1	Repeat 2	Inhibition

No Compound	0.17	0.16	105	95	0
DailyColors Blend,	0.46	0.45	395	385	0
0.0004%
DailyColors Blend,	0.15	0.16	85	95	10
0.002%
DailyColors Blend,	0.07	0.07	5	5	95
0.01%
INCB024360,	0.17	0.17	105	105	0
0.01 μM
INCB024360,	0.14	0.15	75	85	20
0.1 μM
INCB024360,	0.10	0.11	35	45	60
1 μM
Blank	0.06	0.07

NAMPT:

In still another set of experiments, the inventor sought to determine whether the representative compositions had an effect on the enzymatic activities of recombinant human NAMPT using an in vitro enzymatic assay.
Reagents used are shown in Tables 46-47 below and tested as stated unless indicated otherwise.

TABLE 47

	Compound	Stock	Dissolving
Compound I.D.	Supplied	Concentration	Solvent	Test Range

HP Color Blend lot#	Powder		1 % (w/v)	70% EtOH	004%, 0.002%,
33890000X11020				and 0.01%
FK-866*	Powder	10 mM	DMSO	0.001 μM, 0.01μM,
				and 0.1 μM,

TABLE 48

		Enzyme	Enzyme Used
Assay	Catalog #	Lot #	(ng)/Reaction	Substrate

NAMPT	91004	171009-2	100	Nicotinamide 20 μM/
				Phosphoribosyl
				pyrophosphate
40 μM

Assay Conditions: The control compound is dissolved in DMSO. The dilution of the compounds was first performed in 100% DMSO with the highest concentration at 0.01 mM. Each intermediate compound dilution (in 100% DMSO) will then get directly diluted 10×fold into assay buffer for an intermediate dilution of 10% DMSO in assay buffer and 5 μl of the dilution was added to a 50 μl reaction so that the final concentration of DMSO is 1% in the reactions of the control compound and NAMPT only. For NAMPT, the compounds (see 2.2) were preincubated with NAMPT enzyme (see 2.3.1) in for 30 minutes. All of the enzymatic reactions were conducted in duplicate at 30° C. for 120 minutes by adding the substrate mixture containing in a 50 μl mixture containing 50 mM Tris-HCl, pH 8.0, 12.5 mM MgCl2, 20 μM nicotinamide, 40 μM PRPP, 20 μM ATP, 30 μg/mL of alcohol dehydrogenase, 10 μg/mL of NMNAT, 1.5% alcohol, 1 mM DTT, 0.02% BSA, 0.01% Tween 20. The final concentration of DMSO in all reactions was 1%. Fluorescence intensity was measured at an excitation of 340 nm and an emission of 460 nm using a Tecan Infinite M1000 microplate reader.
Data Analysis: NAMPT activity assays were performed in duplicates at each concentration. The fluorescent intensity data were analyzed using the computer software, GraphPad Prism. In the absence of the compound, the fluorescent intensity (Ft) in each data set was defined as 100% activity. In the absence of NAMPT, the fluorescent intensity (Fb) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(F−Fb)/(Ft−Fb), where F=the fluorescent intensity in the presence of the compound. The values of percentage activity were plotted on a bar graph.
Results for the above experiments are shown in Table 48 and FIG. 17. As will be readily appreciated, the representative composition had an inhibitory effect on NAMPT.

TABLE 49

	NAMPT Activity	Background
	(Fluorescence	(Fluorescence
	Intensity)	count)	% Activity

Compound

Repeat

1	Repeat 2	Repeat 1	Repeat 2	Repeat 1	Repeat 2	% Inhibition

No Compound	3507	3493	476	474	100	100	0
33890000X11020, 0.0004%	3512	3494	508	516	99	99	1
33890000X11020, 0.002%	3493	3515	737	715	91	92	8
33890000X11020, 0.01%	3625	3633	1657	1646	65	65	35
FK-866, 0.001 μM	3328	3345	450	442	95	96	4
FK-866, 0.01 μM	1750	1766	454	461	42	43	57
FK-866, 0.1 μM	463	468	440	442	0	0	100
Background	452	460	438	442

PCSK9:

In further experiments, the inventor sought to determine whether the representative compositions had an effect on binding of recombinant human PCSK9 and LDLR using an in vitro ELISA.
The reagents used are shown in Tables 49-50 below and tested as stated unless indicated otherwise (Anti-PCSK9 was used as a reference compound).

TABLE 50

	Form		Dissolving
Sample	Supplied	Stock Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.004, 0.02
lot#33890000X11020				and 0.1%
Anti-PCSK9*	Solution	15 μM	DMSO	0.1, 1 and
				10 nM

TABLE 51

Assay	Catalog #	Lot #	Protein (ng/well)

PCSK9-Biotin	71304	191212-1	100
LDLR	71205	190927-G	50

Assay Conditions: 5 μl of sample or reference inhibitor was pre-incubated with 25 μl of 1× PCSK9 Assay Buffer in assay wells before starting the reaction by the addition of 20 μl of 2.5 ng/μl PCSK9-Biotin. Reaction was progressed for 2 hours at room temperature. Then, wells were washed three times with 1× PCSK9 Assay Buffer and blocked with Blocking Buffer for 10 minutes. 100 μl of Streptavidin-HRP was added to all wells and incubated for 1 hour. Lastly, plate was emptied, washed three times and blocked before the addition of 100 μl of freshly prepared HRP chemiluminescent substrates to every well. Immediately, the luminescence of the samples was measured in a BioTek Synergy 2 microplate reader.
Data Analysis: Binding activity assays were performed in duplicates at each concentration. The luminescence signal was analyzed and compared. In the absence of the compound, the signal in each data set was defined as 100% (Ce) activity. In the absence of ligand (no LDLR), the signal in each data set was defined as 0% (C0) activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C0)/(Ce−C0), where C=the intensity in the presence of the compound (all percent activities below zero were shown zero in the table). Compound fluorescence was removed by subtracting fluorescence at reaction time=0.
Results for the PCSK9 binding inhibition assay are shown in Table 51 and FIG. 18. As can be clearly seen form the data, the representative composition had significant inhibitory activity on binding of recombinant human PCSK9 and LDLR.

TABLE 52

	Signal (Luminescence
	counts)	Activity (%)	Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	78139	77171	101	99
#33890000X11020, 0.0004%	66611	67834	86	87	13
#33890000X11020, 0.002%	63124	64635	81	83	18
#33890000X11020, 0.01%	51675	53641	66	69	32
Anti-PCSK9, 0.1 nM	73367	73719	94	95	5
Anti-PCSK9, 1 nM	50528	52995	65	68	33
Anti-PCSK9, 10 nM	2614	2593	3	3	97
Blank	304	321

CD47:

In additional experiments, the inventor further sought to determine whether the representative compositions had an effect on the binding activity of recombinant human CD47 (hCD47) with human SIRP-α (hSIRP-α) using an in vitro binding assay.
The reagents used are shown in Tables 52-53 below and tested as stated unless indicated otherwise (Anti-PCSK9 was used as a reference compound).

TABLE 53

	Compound	Stock
Compound I.D.	Supplied	Concentration	Test Range

DailyColors blend	Solution		1% in 70%	0.01, 0.002, 00004%
		ethanol
SIRP-α	Solution	2.15 mg/ml	0.05, 0.5, 5 μM

TABLE 54

	Catalog #	Protein
Proteins	(lot#)	Reaction

hCD47, Fc fusion	71177	100 ng
	(140429)
hSIRP-α, Biotin-labeled	71138	600 ng
	(150604)

Assay Conditions: CD47 was coated using 50 μL at 2 ng/μL at 4° C. overnight. After wash and block steps the test compounds were added to CD47-coated plate followed by addition of SIRP-α-biotin. Reaction was incubated for 2 h at room temperature. Binding was detected using HRP-conjugated Streptavidin.
Data Analysis: Binding assays were performed in duplicate at each concentration. The luminescence data were analyzed using the computer software, GraphPad Prism. Percent inhibition was determined by normalizing the data to signal from negative control wells (uncoated wells treated with the biotinylated ligand, set as 100% inhibition) and positive control wells (coated wells treated with the biotinylated ligand in the absence of any inhibitor, set as 0% inhibition).
Results for the above experiment are listed in Table 54 and FIG. 19. Once more, it can be seen that the representative composition had significant inhibitory effect on the binding activity of recombinant human CD47 (hCD47) with human SIRP-α.

TABLE 55

	Luminescence	Activity %	Inhibition

Compound

Repeat

1	Repeat 2	Repeat 1	Repeat 2	%

Negative Control	92	94
Positive Control	36937	38732	98	102	0
0.0004%	36685	38062	97	101	1
DailyColors blend
0.002%	33162	33355	88	88	12
DailyColors blend
0.01% DailyColors	25830	26539	68	70	31
blend
0.05 μM SIRP-α	35347	36581	93	97	5
0.5 μM SIRP-α	19746	19259	52	51	49
5 μM SIRP-α	5310	6891	14	18	84

CD38:

In another set of experiments, the inventor sought to determine whether the representative compositions and fractions thereof as well as a multivitamin mix and other compounds known to affect NAD+ levels had an effect on the hydrolase enzymatic activity of recombinant human CD38 using an in vitro enzymatic assay.
Reagents used are shown in Tables 55-59 below and tested as stated unless indicated otherwise (*Apigenin was used as reference compound). Table 55 shows the representative composition, while Table 56 shows the colored fractions and multivitamin mix. Here, DC-5=Yellow Blend, DC-9=Purple Blend, DC-21-Green Blend, DC-13=Red Blend DCH-TIV 1.0 (Adult Centrum Multivitamins). The red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, and Elderberry as listed above. Table 57 shows two compounds known to influence NAD+ levels: Commercially available “Elysium Health NAD” and “TrueNiagen”, both containing nicotinamide riboside, while Table 58 shows the representative composition (DCH-TIV-0.5) and a multivitamin composition (DCH-TIV-1.0 (Adult Centrum Multivitamin)). Table 59 shows the enzyme and substrate used in this set of experiments.

TABLE 56

	Form		Dissolving
Sample	Supplied	Stock Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.004, 0.02
lot#33890000X11020				and 0.1%
Apigenin	Solution
	50 mM	DMSO				1, 10 and
				100 μM

TABLE 57

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

D5	Powder
	1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D9	Powder
	1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D13	Powder
	1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D21	Powder
	1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
Apigenin	Solution
	50 mM	DMSO			1, 10 and 100 μM

TABLE 58

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-ELY-1.0	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
(Elysium Health				and 0.01%
NAD)
DCH-TN-1.0	Powder	1% (w/v)	70% EtOH	0.0004, 0.002
(TruNiagen;				and 0.01%
nicotinamide riboside)
Apigenin	Solution		50 mM	DMSO	1, 10 and
				100 μM

TABLE 59

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-TIV-0.5	Powder	1% (w/v)	70% EtOH	0.004, 0.02
DCH-TIV-1.0	Powder	1% (w/v)	70% EtOH	and 0.1%
(Adult Centrum				0.004, 0.02
Multivitamin)				and 0.1%
Apigenin	Solution
	50 mM	DMSO			1, 10 and
				100 μM

TABLE 60

			Enzyme Used
Assay	Catalog #	Enzyme Lot #	(ng)/Reaction	Substrate

CD38	71277	170801-1	20	10 μM ε-NAD

Assay Conditions: 10 μl of the sample or reference inhibitor was added to 20 μl of enzyme solution and pre-incubated for 30 minutes. The enzymatic reactions were started by the addition of 20 μl of the substrate ε-NAD+, for a total reaction volume of 50 μl. Reaction time was 10 minutes, and then fluorescence intensity at an excitation of 300 nm and an emission of 410 nm was read using a Tecan Infinite M1000 microplate reader.
Data Analysis: Enzyme activity assays were performed in duplicates at each concentration. The fluorescence intensity data were analyzed and compared. In the absence of the compound, the intensity in each data set was defined as 100% (Ce) activity. In the absence of enzyme, the intensity in each data set was defined as 0% (C0) activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C0)/(Ce−C0), where C=the intensity in the presence of the compound (all percent activities below zero were shown zero in the table). Compound fluorescence was removed by subtracting fluorescence at reaction time=0.
The results for the above substrates are shown in Tables 60-63 and FIGS. 20-23. More specifically, Table 60 and FIG. 20 show the results where the representative composition was used for inhibition of CD38. As can be readily seen from the data, the representative composition exhibited remarkably strong inhibition. Table 61 and FIG. 21 show the results where the colored fractions of the representative composition was used for inhibition of CD38. Notably, here as well a strong inhibition was observed. Moreover, it should be noted that the colored fractions in the representative composition provided a strong synergistic effect with respect to CD38 inhibition.
Table 62 and FIG. 22 show the results for corresponding experiments where the where the “Elysium Health NAD” and “TrueNiagen” (both containing nicotinamide riboside) were used for inhibition of CD38. Here, both formulations showed inhibition of CD38, however, not to the same extent as for the representative composition. In contrast, Table 63 and FIG. 23 show the results for corresponding experiments where a multivitamin composition was used to inhibit CD38. Here, a direct comparison is shown between the representative composition (DCH-TIV-0.5) and the multivitamin composition (DCH-TIV-1.0 in Table 63, DCH-TIG-1.0 in FIG. 23).

TABLE 61

	Net Signal
	(Fluorescence counts)	Activity (%)	Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	4393	4027	104	96
#33890000X11020, 0.0004%	603	694	14	16	85
#33890000X11020, 0.002%	411	516	10	12	89
#33890000X11020, 0.01%	346	311	8	7	92
Apigenin, 1 μM	3461	3761	82	89	14
Apigenin, 10 μM	647	618	15	14	85
Apigenin, 100 μuM	185	135	4	3	96
Blank	3	19	0	0

TABLE 62

	Net Signal
	(Fluorescence counts)	Activity (%)	Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	9204	9171	100	100
D5, 0.0004%	8528	8830	93	96	6
D5, 0.002%	6850	6811	74	74	26
D5, 0.01%	4253	4353	46	47	53
D9, 0.0004%	8679	9582	94	104	1
D9, 0.002%	8327	8618	91	94	8
D9, 0.01%	7801	7614	85	83	16
D13, 0.0004%	4811	5203	52	57	46
D13, 0.002%	5168	5154	56	56	44
D13, 0.01%	3876	3771	42	41	59
D21, 0.0004%	8340	9222	91	100	4
D21, 0.002%	8122	8226	88	90	11
D21, 0.01%	5853	5812	64	63	37
Apigenin, 1 uM	8650	8614	94	94	6
Apigenin, 10 uM	4670	5416	51	59	45
Apigenin, 100 uM	1979	2165	21	23	78
Blank	29	25

TABLE 63

	Net Signal
	(Fluorescence counts)	Activity (%)	Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	13504	13458	100	100	0
DCH-ELY-1,0, 0.01%	1736	1733	13	13	87
DCH-ELY-1.0, 0.002%	4571	4564	34	34	66
DCH-ELY-1.0, 0.0004%	8806	8767	65	65	35
DCH-TN-1.0, 0.01%	1742	1722	13	13	87
DCH-TN-1.0, 0.002%	3776	3716	28	27	72
DCH-TN-1.0, 0.0004%	7650	7587	57	56	44
Apigenin, 1 μM	11531	11647	86	86	14
Apigenin, 10 μM	7118	7228	53	54	47
Apigenin, 100 μM	5012	5473	37	40	61
Blank	30	29

TABLE 64

	Net Signal		Inhi-
	(Fluorescence counts)	Activity (%)	bition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	4383	4355	100	100	0
DCH-TIV-0.5, 0.0004%	838	810	18	18	82
DCH-TIV-0.5, 0.002%	653	541	14	12	87
DCH-TIV-0.5, 0.01%	240	296	5	6	95
DCH-TIV-1.0, 0.0004%	4371	4320	100	99	1
DCH-TIV-1.0, 0.002%	4168	4209	95	96	4
DCH-TIV-1.0, 0.01%	3812	3744	87	86	14
Apigenin, 1 μM	4138	3883	95	89	8
Apigenin, 10 μM	3091	3057	70	70	30
Apigenin, 100 μM	632	728	14	16	85
Blank	52	23

JAK1/JAK2/JAK3:

In further experiments, the inventor sought to determine whether the representative compositions and fractions thereof had an effect on the enzymatic activities of the recombinant human kinases JAK1, JAK2, and JAK3 using an in vitro enzymatic assay.
Reagents used are shown in Tables 64-66 below and tested as stated unless indicated otherwise (*Apigenin was used as reference compound). Table 64 shows the representative composition, while Table 65 shows the colored fractions. Here, DC-5=Yellow Blend, DC-9=Purple Blend, DC-21-Green Blend, DC-13=Red Blend. The red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, and Elderberry as listed above. Staurosporine was used as a reference compound. Table 66 lists the enzymes and substrates used in the assays.

TABLE 65

	Com-	Stock			Inter-
	pound	Con-	Dissolving	Test	mediate
Compound I.D.	Supplied	centration	Solvent	Range	Dilution

DailyColors Blend	solution		1% (w/v)	70%	0.01%,	Water
Lot #			EtOH	0.002%,
33890000X11020				0.0004%
Staurosporine	solution
	1 mM	DMSO	0.001 μM,	10%
				0.01 μM,	DMSO
				0.1 μM	(aq)

TABLE 66

Com	Com-	Stock			Inter-
pound	pound	Con-	Dissolving		mediate
I.D.	Supplied	centration	Solvent	Test Range	Dilution

DC-5	solution	1% (w/v)	70% EtOH	0.01%,	Water
				0.002%,
				0.0004%
DC-9	solution	1% (w/v)	70% EtOH	0.01%,	Water
				0.002%,
				0.0004%
DC-13	solution	1% (w/v)	70% EtOH	0.01%,	Water
				0.002%,
				0.0004%
DC-21	solution	1% (w/v)	70% EtOH	0.01%,	Water
				0.002%,
				0.0004%
Staurosporine	solution
	1 mM	DMSO		30 nM-3 μM	10%
					DMSO
					(aq)

TABLE 67

	Catalog #	Enzyme Used (ng)/
Assay	(Lot #)	Reaction	Substrate

Jak1	40449	100	0.1 mg/ml, IRS1-tide/
	(190919-3)		10 μM ATP
Jak
2	40450	50	0.2 mg/ml Poly (Glu, Tyr)/
	(190603-G)		10 μM ATP
Jak 3	40452	10	0.2 mg/ml Poly (Glu, Tyr)/
	(150921-B2)		10 μM ATP

Assay Conditions: The assay was performed using Kinase-Glo Plus luminescence kinase assay kit (Promega). It measures kinase activity by quantitating the amount of ATP remaining in solution following a kinase reaction. The luminescent signal from the assay is correlated with the amount of ATP present and is inversely correlated with the amount of kinase activity. The reference compound was diluted as noted. The compound was diluted in water and 5 μl of the dilution was added to a 50 μl reaction. All of the enzymatic reactions were conducted at 30° C. for 45 minutes. The 50 μl reaction mixture contains 40 mM Tris, pH 7.4, 10 mM MgCl2, 0.1 mg/ml BSA, 1 mM DTT, 10 μM ATP, Kinase substrate and the respective enzyme. After the enzymatic reaction, 50 μl of Kinase-Glo Plus Luminescence kinase assay solution (Promega) was added to each reaction and incubate the plate for 15 minutes at room temperature. Luminescence signal was measured using a BioTek Synergy 2 microplate reader.
Data Analysis: Kinase activity assays were performed in duplicate at each concentration. The luminescence data were analyzed using the computer software, GraphPad Prism. The difference between luminescence intensities in the absence of Kinase (Lut) and in the presence of Kinase (Luc) was defined as 100% activity (Lut−Luc). Using luminescence signal (Lu) in the presence of the compound, % activity was calculated as: % activity={(Lut−Lu)/(Lut−Luc)}×100%, where Lu=the luminescence intensity in the presence of the compound (all percent activities below zero were shown zero in the table).
The results for inhibition using the representative composition are shown in Tables 67-69 and FIGS. 24-26. Table 67 and FIG. 24 show results for JAK1 inhibition using the representative composition. Table 68 and FIG. 25 show results for JAK2 inhibition using the representative composition. Table 69 and FIG. 26 show results for JAK3 inhibition using the representative composition. As can be readily taken from these results, the inhibition of all three tested JAK kinases was significant and substantial, matching or exceeding the inhibition provided by the reference compound.

TABLE 68

	Kinase Activity		%
	Luminescence	% Activity	Inhi-

Compounds	Repeat1	Repeat2	Repeat1	Repeat2	bition

No Compound	24012	26135	107	93
0.0004% DailyColors Blend	25706	26483	96	91	6
0.002% DailyColors Blend	37313	38359	23	16	81
0.01% DailyColors Blend	42256	42457	0	0	100
0.001 μM Staurosporine	25176	27054	99	87	7
0.01 μM Staurosporine	37642	35358	21	35	72
0.1 μM Staurosporine	40161	40781	5	1	97
Background	40868	40953

TABLE 69

	Kinase Activity		%
	Luminescence	% Activity	Inhi-

Compounds	Repeat1	Repeat2	Repeat1	Repeat2	bition

No Compound	20027	20151	100	100
0.0004% DailyColors Blend	35932	34651	21	27	76
0.002% DailyColors Blend	36319	36342	19	19	81
0.01% DailyColors Blend	41665	40587	0	0	100
0.001 μM Staurosporine	19100	20922	105	96	0
0.01 μM Staurosporine	36988	36554	16	18	83
0.1 μM Staurosporine	42700	40639	0	0	100
Background	40021	40218

TABLE 70

	Kinase Activity		%
	Luminescence	% Activity	Inhi-

Compounds	Repeat1	Repeat2	Repeat1	Repeat2	bition

No Compound	18990	20947	100	95
0.0004% DailyColors Blend	35140	35074	20	21	79
0.002% DailyColors Blend	36502	37407	13	8	89
0.01% DailyColors Blend	39437	41388	0	0	100
0.001 μM Staurosporine	23814	23348	80	82	19
0.01 μM Staurosporine	32137	32417	36	35	65
0.1 μM Staurosporine	39970	40121	0	0	100
Background	38815	39237

Tables 70-72 and FIGS. 27-29 show corresponding results for the colored fractions. Here, Table 70 shows results for JAK1 inhibition using colored fractions of the representative composition. Table 71 shows results for JAK2 inhibition using colored fractions of the representative composition, and Table 72 shows results for JAK3 inhibition using colored fractions of the representative composition. Table 73 is a summary table of the results in Tables 70-72. Notably, a synergistic effect on inhibition against all three JAK kinases was observed at high concentrations where all colored fractions were used together in the representative composition as compared to individual colored fractions. Moreover, it should once more be noted that the compositions presented herein had similar inhibitory properties as compared to the reference compound.

TABLE 71

	Kinase Activity
	Luminescence	% Activity

Compounds	Repeat1	Repeat2	Repeat1	Repeat2

No Compound	19821	17710	95	105
DC-5, 0.0004%	20319	18662	92	101
DC-5, 0.002%	21569	20952	86	89
DC-5, 0.01%	27118	24076	59	74
DC-9, 0.0004%	22093	20295	84	93
DC-9, 0.002%	31746	29312	37	49
DC-9, 0.01%	37123	35475	10	18
DC-13, 0.0004%	27065	25415	59	68
DC-13, 0.002%	37759	34432	7	24
DC-13, 0.01%	36702	35308	12	19
DC-21, 0.0004%	16635	19342	110	97
DC-21, 0.002%	17067	20286	108	93
DC-21, 0.01%	23118	25700	79	66
Staurosporine, 1 nM	21164	24810	88	70
Staurosporine, 10 nM	28640	32000	52	35
Staurosporine, 100 nM	36908	39509	11	0
Background	39443	39051

TABLE 72

	Kinase Activity
	Luminescence	% Activity

Compounds	Repeat1	Repeat2	Repeat1	Repeat2

No Compound	5301	6612	102	98
DC-5, 0.0004%	6760	6751	98	98
DC-5, 0.002%	7887	9950	94	88
DC-5, 0.01%	9442	8832	89	91
DC-9, 0.0004%	17918	16309	63	68
DC-9, 0.002%	33289	32043	16	20
DC-9, 0.01%	34796	35989	12	8
DC-13, 0.0004%	30946	29748	24	27
DC-13, 0.002%	35743	34642	9	12
DC-13, 0.01%	36184	36167	7	8
DC-21, 0.0004%	6344	8335	99	93
DC-21, 0.002%	7862	9809	94	88
DC-21, 0.01%	13577	16488	77	68
Staurosporine, 1 nM	9482	10430	89	86
Staurosporine, 10 nM	17529	16180	65	69
Staurosporine, 100 nM	34660	34762	12	12
Background	39026	38243

TABLE 73

	Kinase Activity
	Luminescence	% Activity

Compounds	Repeat1	Repeat2	Repeat1	Repeat2

No Compound	16444	18933	106	94
DC-5, 0.0004%	18578	17845	96	99
DC-5, 0.002%	19889	20404	89	87
DC-5, 0.01%	22322	22047	78	79
DC-9, 0.0004%	30974	32880	37	27
DC-9, 0.002%	34085	35077	22	17
DC-9, 0.01%	34611	35273	19	16
DC-13, 0.0004%	34002	34554	22	19
DC-13, 0.002%	35870	36421	13	11
DC-13, 0.01%	36418	36584	11	10
DC-21, 0.0004%	16774	19477	104	91
DC-21, 0.002%	18739	21558	95	82
DC-21, 0.01%	23945	29444	70	44
Staurosporine, 1 nM	21050	24827	84	66
Staurosporine, 10 nM	32876	34538	27	20
Staurosporine, 100 nM	37496	39233	5	0
Background	39074	38166

	TABLE 74

		% Inhibition

	Compounds	JAK1	JAK2	JAK3

DC-5, 0.0004%	4	2	2
DC-5, 0.002%	12	9	12
DC-5, 0.01%	33	10	21
DC-9, 0.0004%	12	34	68
DC-9, 0.002%	57	82	81
DC-9, 0.01%	86	90	82
DC-13, 0.0004%	36	75	79
DC-13, 0.002%	85	89	88
DC-13, 0.01%	84	92	90
DC-21, 0.0004%	0	4	2
DC-21, 0.002%	0	9	12
DC-21, 0.01%	28	28	43
Staurosporine, 1 nM	21	12	25
Staurosporine, 10 nM	56	33	77
Staurosporine, 100 nM	94	88	97

CD39:

In further experiments, the inventor sought to determine whether the representative compositions had an effect on the enzymatic activity of recombinant human CD39 using an in vitro enzymatic assay.
Reagents used are shown in Tables 74-77 below and tested as stated unless indicated otherwise (*POM-1 was used as reference compound). Table 74 shows the representative composition at standard concentrations, and Table 75 shows the representative composition at low concentrations. Table 76 shows colored fractions of the representative composition at standard concentrations. Here, D5 is the Yellow Blend, D9 is the Purple Blend, D21 is the Green Blend, D13 is the Red Blend, and D31 is CBD. As before, the red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, Elderberry. Table 77 lists CD39.

TABLE 75

	Form	Stock	Dissolving
Compound	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.0004, 0.002
lot#33890000X11020				and 0.01%
POM-1*	Powder		1 mM	Water	0.001, 0.01
				and 0.1 μM

TABLE 76

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder		1% (w/v)	70% EtOH	0.00000046,
lot#33890000X11020				0.000014,
				0.00004,
				0.00013, and
				0.0004%
POM-1*	Powder		1 mM	Water	0.001, 0.01
				and 0.1 μM

TABLE 77

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

D5	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D9	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D13	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D21	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D31	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
POM-1*	Powder	1 mM	Water	0.001, 0.01 and 0.1 μM
AMPCP*	Powder	100 mM	Water			1, 10 and 100 μM

TABLE 78

		Enzyme	Enzyme Used
Assay	Catalog #	Lot #	(ng)/Reaction	Substrate

CD39	71284	180406	10	70 μM ATP

Assay Conditions: In general, all assays points were done by following the CD39 and CD73 Inhibitor Screening Assay Kit protocol (BPS Bioscience, #79278 and 72055, respectively). The CD39 enzymatic reactions were conducted in duplicate at room temperature for 30 minutes in a 50 μl mixture containing assay buffer, ATP, CD39 enzymes, and the test compound. Test compounds were preincubated with the enzyme for 30 minutes. Reactions were started by addition of the substrate. The 50μl reactions were carried out in a 96-well transparent plate. After enzymatic reactions, 100μl of Colorimetric Detection Reagent was added to the reaction mix. After a 15 minutes incubation, absorbance was measured using a Tecan plate reader at 630 nm.
Data Analysis: Enzyme activity assays were performed in duplicates at each concentration. The Absorbance intensity data were analyzed and compared. In the absence of the compound, the intensity in each data set was defined as 100% (Ce) activity. In the absence of enzyme, the intensity in each data set was defined as 0% (C0) activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C0)/(Ce−C0), where C=the intensity in the presence of the compound (all percent activities below zero were shown zero in the table).
Remarkably high and significant inhibitory activity was found for CD39 across all tested concentrations as is shown in Tables 77-78 below, with Table 78 and FIG. 27 depicting the results for standard concentrations and Table 79 and FIG. 28 showing results for low concentrations. As can be readily seen from the results, inhibition relative to the reference inhibitor was unexpectedly strong relative to known reference inhibitor POM-1. Notably, the IC50 concentration for the composition was at 0.000044%. When tested for inhibitory activity for the colored fractions, the inhibitory activity partitioned partially, but not completely, to selected fractions as can be seen from the results in Table 80 and FIG. 29.

TABLE 79

	Net absorbance	Activity (%)	CD39

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	Inhibition (%)

No compound	0.51	0.55	95	105
HP Color Blend, 0.0004%	0.01	0.01	1	1	99
HP Color Blend, 0.002%	0.01	0.01	2	2	98
HP Color Blend, 0.01%	0.02	0.01	3	2	98
POM-1, 0.001 μM	0.42	0.41	79	78	21
POM-1, 0.01 μM	0.26	0.30	49	56	47
POM-1,0.1 μM	0.08	0.10	14	19	84
Blank	0.00	0.00

TABLE 80

	Net absorbance	Activity (%)	Inhibition

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	(%)

No compound	0.289	0.277	102	98
HP Color Blend, 0.0000046%	0.247	0.227	87	80	16
HP Color Blend, 0.000014%	0.236	0.226	83	80	18
HP Color Blend, 0.000044%	0.138	0.149	48	53	49
HP Color Blend, 0.00013%	0.040	0.043	14	15	85
HP Color Blend, 0.0004%	0.025	0.024	8	8	92
POM-1, 0.001 μM	0.281	0.254	99	90	5
POM-1, 0.01 μM	0.154	0.145	54	51	47
POM-1, 0.1 μM	0.073	0.073	26	26	74
Blank	0.001	0.001

TABLE 81

	Net absorbance	Activity (%)	Inhibition

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	(%)

No compound	0.54	0.53	100	100
D5, 0.0004%	0.48	0.44	88	81	15
D5, 0.002%	0.48	0.44	90	83	14
D5, 0.01%	0.48	0.48	90	89	11
D9, 0.0004%	0.43	0.41	80	77	21
D9, 0.002%	0.30	0.31	56	57	44
D9, 0.01%	0.20	0.21	37	38	62
D13, 0.0004%	0.01	0.01	1	1	99
D13, 0.002%	0.00	0.01	0	2	99
D13, 0.01%	0.01	0.00	1	0	99
D21,0.0004%	0.26	0.26	49	49	51
D21, 0.002%	0.23	0.25	42	46	56
D21, 0.01%	0.25	0.22	47	40	57
D31,0.0004%	0.50	0.49	93	91	8
D31, 0.002%	0.48	0.49	90	92	9
D31, 0.01%	0.52	0.55	96	103	1
POM-1, 0.001 μM	0.51	0.49	96	92	6
POM-1, 0.01 μM	0.19	0.20	36	37	64
POM-1,0.1 μM	0.08	0.06	16	10	87
Blank	0.00	0.00

The inventor then further investigated whether one or more specific plant materials and their polyphenols were associated with the inhibitory activity against CD39. To that end, the inventor tested two components of the red colored blend: Apple Extract (DCH-IC50X) and Pomegranate extract (DCH-IC50Y) at the low concentration ranges as shown in Table 81 with otherwise identical assay conditions. Results are shown in Table 82 and FIG. 30.

TABLE 82

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-IC50X (Apple	Powder	1% (w/v)	70% EtOH	0.0000046, 0.000014,
Extract of RED Blend)				0.00004, 0.00013,
DCH-IC50Y	Powder		1% (w/v)	70% EtOH	and 0.0004%
(Pomegranate Extract
of RED Blend)
POM-1*	Powder	1 mM	Water	0.001, 0.01 and 0.1 μM

TABLE 83

	Net absorbance	Activity (%)	Inhibition

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	(%)

No compound	0.30	0.29	102	98	0
DCH-IC50X, 0.0000046%	0.28	0.28	95	95	5
DCH-IC50X, 0.000014%	0.28	0.27	97	92	6
DCH-IC50X, 0.00004%	0.27	0.27	92	91	8
DCH-IC50X, 0.00013%	0.27	0.26	91	88	11
DCH-IC50X, 0.0004%	0.25	0.26	85	88	13
DCH-IC50Y, 0.0000046%	0.07	0.07	22	24	77
DCH-IC50Y, 0.000014%	0.03	0.03	8	8	92
DCH-IC50Y, 0.00004%	0.03	0.03	7	8	92
DCH-IC50Y, 0.00013%	0.03	0.03	8	9	91
DCH-IC50Y, 0.0004%	0.03	0.03	9	10	91
POM-1, 0.001 μM	0.25	0.26	86	89	13
POM-1, 0.01 μM	0.20	0.19	67	66	33
POM-1, 0.1 μM	0.09	0.09	30	29	70
Blank	0.01	0.00

In still further experiments, the inventor also investigated whether CD39 could also be inhibited by a multivitamin mix. To that end, a comparative experiment was conducted between a multivitamin mix (denoted as DCH-TIV-1.0 (Adult Centrum Multivitamin)) and the representative composition (denoted as DCH-TIC-0.5) using the same experimental procedure for CD39 as described above. The compositions are shown in Table 83.

TABLE 84

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-TIC-0.5	Powder	1% (w/v)	70% EtOH	0.004, 0.02 and 0.1%
DCH-TIV-1.0	Powder	1% (w/v)	70% EtOH	0.004, 0.02 and 0.1%
(Adult Centrum
Multivitamin)
POM-1*	Solution	1 mM	Water	0.001, 0.01 and 0.1 μM

The results for this comparison are shown in Table 84 and FIG. 31. As can be clearly seen form the results, the representative composition had very strong inhibitory effect on CD39 whereas the multivitamin composition had substantially no significant inhibitory effect.

TABLE 85

	Net Signal

(Absorbance)

Activity (%)

Inhibition

Condition	Repeat 1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	0.32	0.32	99	101	0
DCH-TIC-0.5, 0.0004%	0.04	0.04	9	10	90
DCH-TIC-0.5, 0.002%	0.04	0.04	10	9	91
DCH-TIC-0.5, 0.01%	0.05	0.05	15	15	85
DCH-TIV-1.0, 0.0004%	0.30	0.30	93	92	8
DCH-TIV-1.0, 0.002%	0.30	0.29	92	91	8
DCH-TIV-1.0, 0.01%	0.31	0.31	96	98	3
POM-1, 0.001 μM	0.27	0.29	85	89	13
POM-1, 0.01 μM	0.21	0.22	65	68	34
POM-1, 0.1 μM	0.13	0.13	39	38	61
Blank	0.01	0.01

CD73:

In still further experiments, the inventor further sought to determine whether the representative compositions had an effect on the enzymatic activity of recombinant human CD73 using an in vitro enzymatic assay.
Reagents used are shown in Tables 85-89 below and tested as stated unless indicated otherwise (*AMPCP or Quercetin were used as reference compound). Table 85 shows the representative composition at standard concentrations, and Table 86 shows colored fractions of the representative composition at standard concentrations. Here, D5 is the Yellow Blend, D9 is the Purple Blend, D21 is the Green Blend, D13 is the Red Blend, and D31 is CBD. As before, the red blend included Apple Extract, Pomegranate Extract, Tomato Powder, Beet; the green blend included Olive Extract, Rosemary Extract, Green Coffee Bean Extract, Kale; the orange/yellow blend included Onion Extract, Ginger Extract, Grapefruit Extract, Carrot; and the purple/blue blend included Grape, Blueberry Extract, Currant, Elderberry. Table 87 lists CD73.

TABLE 86

	Form	Stock	Dissolving
Compound	Supplied	Conc.	Solvent	Test Range

HP Color Blend	Powder	1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
lot #33890000X11020
Quercetin*	Powder	100 mM	Water		1, 10 and 100 μM

TABLE 87

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

D5	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D9	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D13	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D21	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
D31	Powder		1% (w/v)	70% EtOH	0.0004, 0.002 and 0.01%
AMPCP*	Powder	100 mM	Water			1, 10 and 100 μM

TABLE 88

		Enzyme	Enzyme Used
Assay	Catalog #	Lot #	(ng)/Reaction	Substrate

CD73	71184	190123	3	100 μM ADP

Assay Conditions: In general, all assays points were done by following the CD39 and CD73 Inhibitor Screening Assay Kit protocol (BPS Bioscience, #79278 and 72055, respectively). The CD73 enzymatic reactions were conducted in duplicate at room temperature for 30 minutes in a 50 μl mixture containing assay buffer, ADP, CD73 enzymes, and the test compound. Test compounds were preincubated with the enzyme for 30 minutes. Reactions were started by addition of the substrate. The 50 μl reactions were carried out in a 96-well transparent plate. After enzymatic reactions, 100 μl of Colorimetric Detection Reagent was added to the reaction mix. After a 15 minutes incubation, absorbance was measured using a Tecan plate reader at 630 nm.
Data Analysis: Enzyme activity assays were performed in duplicates at each concentration. The Absorbance intensity data were analyzed and compared. In the absence of the compound, the intensity in each data set was defined as 100% (Ce) activity. In the absence of enzyme, the intensity in each data set was defined as 0% (C0) activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(C−C0)/(Ce−C0), where C=the intensity in the presence of the compound (all percent activities below zero were shown zero in the table).
As can be see form the results in Tables 88-90, the inhibition of CD73 by the representative composition and its fractions was remarkably high, especially in comparison to the current reference standard. Table 88 and FIG. 32 show the results for CD73 inhibition at standard concentrations and Table 89 and FIG. 33 show the results for CD73 inhibition at low concentrations. Here, the IC50 of the representative composition is at about 0.000044%. Moreover, as can be taken from these results and the results for the colored fractions as shown in Table 90 and FIG. 34, a strong synergy of the colored fractions when used in combination (as in the representative composition) was observed with respect to CD73 inhibition.

TABLE 89

	Net absorbance	Activity (%)	CD73

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	Inhibition (%)

No compound	0.44	0.43	101	99
HP Color Blend, 0.0004%	0.06	0.06	14	14	86
HP Color Blend, 0.002%	0.03	0.04	7	9	92
HP Color Blend, 0.01%	0.01	0.00	2	1	99
Quercetin, 1 μM	0.40	0.36	92	82	13
Quercetin, 10 μM	0.23	0.25	53	58	44
Quercetin, 100 μM	0.12	0.08	27	17	78
Blank	0.00	0.00

TABLE 90

	Net absorbance	Activity (%)	Inhibition

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	(%)

No compound	0.408	0.395	102	98
HP Color Blend, 0.0000046%	0.389	0.388	97	97	3
HP Color Blend, 0.000014%	0.301	0.316	75	79	23
HP Color Blend, 0.000044%	0.169	0.150	42	37	60
HP Color Blend, 0.00013%	0.117	0.153	29	38	67
HP Color Blend, 0.0004%	0.104	0.083	26	20	77
AMPCP, 1 μM	0.368	0.383	92	96	6
AMPCP, 10 μM	0.298	0.306	74	76	25
AMPCP, 100 μM	0.071	0.089	17	22	80
Blank	0.001	0.002

TABLE 91

	Net absorbance	Activity (%)	Inhibition

Condition	Rep. 1	Rep.2	Rep. 1	Rep.2	(%)

No compound	0.33	0.34	99	101
D5, 0.0004%	0.34	0.33	101	97	1
D5, 0.002%	0.33	0.32	100	94	3
D5, 0.01%	0.32	0.32	95	94	5
D9, 0.0004%	0.30	0.27	88	82	15
D9, 0.002%	0.17	0.17	49	51	50
D9, 0.01%	0.01	0.02	3	6	95
D13, 0.0004%	0.28	0.26	83	77	20
D13, 0.002%	0.17	0.19	51	57	46
D13, 0.01%	0.02	0.02	6	5	94
D21, 0.0004%	0.33	0.32	97	96	3
D21, 0.002%	0.33	0.31	98	92	5
D21, 0.01%	0.33	0.34	97	102	1
D31,0.0004%	0.35	0.34	104	101	0
D31, 0.002%	0.35	0.34	103	100	0
D31, 0.01%	0.35	0.33	106	98	0
AMPCP, 1 μM	0.30	0.30	89	89	11
AMPCP, 10 μM	0.18	0.20	52	59	45
AMPCP, 100 μM	0.04	0.04	13	12	88
Blank	0.01	0.00

To further investigate whether CD73 could also be inhibited by a multivitamin formulation, the inventor performed comparative experiments between the representative composition and a multivitamin composition using the same test procedure as outlined above. Table 91 lists the reagents used in this experiment (DCH-TIV-0.5 denotes the representative composition, and DCH-TIV-1.0 denotes Adult Centrum Multivitamin).

TABLE 92

	Form	Stock	Dissolving
Sample	Supplied	Conc.	Solvent	Test Range

DCH-TIV-0.5	Powder	1% (w/v)	70% EtOH	0.004, 0.02 and 0.1%
DCH-TIV-1.0	Powder	1% (w/v)	70% EtOH	0.004, 0.02 and 0.1%
(Adult Centrum
Multivitamin)
AMPCP*	Powder	10 mM	Water	0.1, 1 and 10 μM

As can be readily seen form the results in Table 92 and FIG. 35 the representative composition had substantial inhibitory activity with regard to CD73, however, only minor inhibitory activity with the multivitamin composition was observed.

TABLE 93

	Net Signal

(Absorbance)

Activity (%)

Inhibition

Condition

Repeat

1	Repeat 2	Repeat 1	Repeat 2	(%)

No compound	0.18	0.17	101	99	0
DCH-TIV-0.5, 0.0004%	0.02	0.02	10	11	89
DCH-TIV-0.5, 0.002%	0.01	0.01	4	5	96
DCH-TIV-0.5, 0.01%	0.02	0.02	10	10	90
DCH-TIV-1.0, 0.0004%	0.17	0.17	97	95	4
DCH-TIV-1.0, 0.002%	0.17	0.16	93	92	7
DCH-TIV-1.0, 0.01%	0.20	0.18	112	103	0
AMPCP, 0.1 μM	0.14	0.17	81	96	12
AMPCP, 1 μM	0.09	0.11	53	64	41
AMPCP, 10 μM	0.04	0.05	20	28	76
Blank	0.00	0.00

In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, reaction conditions, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein.
As used herein, the term “administering” a pharmaceutical or nutraceutical composition refers to both direct and indirect administration of the pharmaceutical or nutraceutical composition, wherein direct administration of the pharmaceutical or nutraceutical composition is typically performed by a health care professional (e.g., physician, nurse, dietitian, etc.), and wherein indirect administration includes a step of providing or making available the pharmaceutical or nutraceutical composition to the health care professional or individual in need thereof for direct administration (e.g., via injection, infusion, oral delivery, topical delivery, etc.). It should further be noted that the terms “prognosing” or “predicting” a condition, a susceptibility for development of a disease, or a response to an intended treatment is meant to cover the act of predicting or the prediction (but not treatment or diagnosis of) the condition, susceptibility and/or response, including the rate of progression, improvement, and/or duration of the condition in a subject.
All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.
As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise. As also used herein, and unless the context dictates otherwise, the term “coupled to” is intended to include both direct coupling (in which two elements that are coupled to each other contact each other) and indirect coupling (in which at least one additional element is located between the two elements). Therefore, the terms “coupled to” and “coupled with” are used synonymously.
It should be apparent to those skilled in the art that many more modifications besides those already described are possible without departing from the inventive concepts herein. The inventive subject matter, therefore, is not to be restricted except in the scope of the appended claims. Moreover, in interpreting both the specification and the claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, the terms “comprises” and “comprising” should be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that the referenced elements, components, or steps may be present, or utilized, or combined with other elements, components, or steps that are not expressly referenced. Where the specification or claims refer to at least one of something selected from the group consisting of A, B, C . . . and N, the text should be interpreted as requiring only one element from the group, not A plus N, or B plus N, etc.

Claims

What is claimed is:

1. A method of reducing an NAD⁺ decrease in an individual, comprising administering to the individual a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color, wherein the combination synergistically inhibits CD38.

2. The method of claim 1 wherein the polyphenols are provided in form of the plant materials.

3. The method of claim 2 wherein the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.

4. The method of claim 1 wherein the polyphenols comprise at least one organic acid as listed in Table 2, at least one phenolic compound as listed in Table 3, at least one flavonoid as listed in Table 4, at least one anthocyanin as listed in Table 5, at least one chlorogenic acid as listed in Table 6, at least one betacyanin as listed in Table 7, and/or at least one amino acid/alkaloid as listed in Table 8.

5. The method of claim 1 wherein the step of contacting comprises oral administration to a mammal.

6. The method of claim 1 wherein the plurality of chemically distinct polyphenols are administered at a daily dosage of between 50 and 1,000 mg.

7. The method of claim 1 further comprising a step of co-administering to the mammal a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic.

8. The method of claim 1 wherein the synergistic combination of polyphenols further inhibit at least one enzyme selected from the group consisting of BACE1, CD73, CDK5, JAK1, JAK2, JAK3, ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S.

9. The method of claim 1 wherein the synergistic combination of polyphenols further inhibit at least two enzymes selected from the group consisting of BACE1, CD73, CDK5, JAK1, JAK2, JAK3, ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S.

10. The method of claim 1 wherein the synergistic combination of polyphenols further inhibit at least three enzymes selected from the group consisting of BACE1, CD73, CDK5, JAK1, JAK2, JAK3, ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S.

11. The method of claim 1 wherein administration of the synergistic combination of polyphenols reduces conditions associated with CD38 overactivity.

12. The method of claim 1 wherein administration of the synergistic combination of polyphenols supports energy metabolism and/or immunity.

13. A composition for reducing an NAD⁺ decrease in an individual, comprising a synergistic combination of polyphenols from plant materials having a red color, a green color, an orange-yellow color, and a purple-blue color, wherein the combination synergistically inhibits CD38.

14. The composition of claim 13 wherein the polyphenols are provided in form of the plant materials.

15. The composition of claim 13 or claim 14 wherein the red colored plant materials are selected from the group consisting of an apple extract, a pomegranate extract, a tomato powder, and a beet root powder, wherein the green colored plant materials are selected from the group consisting of an olive extract, a rosemary extract, a green coffee bean extract, and a kale powder, wherein the orange-yellow colored plant materials are selected from the group consisting of an onion extract, a ginger extract, a grapefruit extract, and a carrot powder, and wherein the purple-blue colored plant materials are selected from the group consisting of a grape extract, a blueberry extract, a currant powder, and an elderberry powder.

16. The composition of claim 13 wherein the polyphenols comprise at least one organic acid as listed in Table 2, at least one phenolic compound as listed in Table 3, at least one flavonoid as listed in Table 4, at least one anthocyanin as listed in Table 5, at least one chlorogenic acid as listed in Table 6, at least one betacyanin as listed in Table 7, and/or at least one amino acid/alkaloid as listed in Table 8.

17. The composition of claim 13 further comprising a vitamin, a dietary trace element or mineral, a nicotinamide riboside, a probiotic, and/or a prebiotic.

18. The composition of claim 13 wherein the synergistic combination of polyphenols further inhibits at least one enzyme selected from the group consisting of BACE1, CD73, CDK5, JAK1, JAK2, JAK3, ARG-1, ARG-2, SIRT-1, CD39, IDO1, IDO2, NAMPT, PCSK9, CD47, and Cathepsin S.

19. The composition of claim 13 wherein administration of the synergistic combination of polyphenols reduces conditions associated with CD38 overactivity.

20. The composition of claim 13 wherein administration of the synergistic combination of polyphenols supports energy metabolism and/or immunity.