Patent Application
20170000703
The present technology relates to cosmetic and pharmaceutical compositions useful for controlling the rate of cell destruction and minimizing the appearance of aging. In particular, the present technology relates to compositions and methods comprising a combination of a sirtuin activator with a sirtuin-offsetting agent (also referred to herein as a sirtuin inhibitor).
Sirtuins are a class of protein deacetylase enzymes that require nicotaminde adenine dinucleotide (NAD) as a cofactor in deacetylating lysine residues in target proteins. Acetylation and deacetylation of certain amino acids is a post-translational modification that controls the activities of some target proteins. The targets that were initially discovered were the histone proteins that package DNA in cell nuclei; thus, these enzymes are commonly referred to as histone deaceteylases (HDACs), despite the fact that many non-histone target proteins have been discovered subsequent to the initial discoveries.
Sirtuins have been found to influence various biological phenomena, including cellular stress responses such as DNA repair, replicative senescence and apoptosis (suicide or “programmed cell death” response that cells typically undergo following serious or irreparable damage). Specifically, sirtuins have been found to delay apoptosis of damaged cells, thereby slowing down or eliminating their destruction.
There is concern that the mechanism through which sirtuins may slow down aging may possess an inherently dangerous side effect—specifically, the apoptic response protects organisms by eliminating damaged cells that could otherwise become genetically unstable and lose normal growth controls and proper differentiated function. By having the effect of delaying apoptosis of damaged cells, sirtuins might allow them to escape normal checks, resulting in overgrowth of tissues with abnormal progeny cells, which would compromise proper tissue function and possibly even imperil organism survival.
Thus, a need exists for “safe” sirtuin compounds and methods—specifically, compositions that harness the anti-aging benefits of sirtuins without the undesirable side effects; as well as methods of optimizing such compositions and of using them in anti-aging applications for patients.
In certain embodiments, the present technology is directed to compositions comprising a sirtuin activator (also referred to herein as a sirtuin stimulator) as well as a sirtuin-offsetting agent. (also referred to herein as a sirtuin inhibitor).
In certain embodiments, the present technology is directed to methods of formulating a composition, or of optimizing the efficacy of a composition for a patient, or of optimizing cell maintenance in a patient, the methods comprising selecting a sirtuin stimulator having a known quantitative prolonging effect on a cell, and selecting a sirtuin-offsetting agent having a known quantitative opposite effect on the cell, and optimizing the balance between the two effects based on a known desired ultimate effect on the cell.
In certain embodiments, the present technology is directed to a method of prolonging the life of a cell and simultaneously avoiding proliferation of cell damage, the method comprising the steps of: stimulating Sirt1 activity in the cell; and inhibiting a non-sirtuin HDAC in the cell.
In certain embodiments, the present technology is directed to methods of treating a patient, or methods of reducing the appearance of aging in a patient, or methods of optimizing cell maintenance in a patient, comprising applying a composition including sirtuin stimulator and a sirtuin-offsetting agent to the body of a patient.
Sirtuin function appears to be affected by metabolic state. More broadly, sirtuins appear to help individual cells (and therefore organisms overall) to survive stress, likely by effecting a delay in the apoptotic response, so as to allow cells the time and opportunity to repair whatever stress-related damage they may suffer. It is presumed that any lengthening of lifetime of a cell that results from enhanced sirtuin activity is a consequence of delayed apoptosis of cells under stressful conditions. The cells of greatest interest in this regard would be stem cells. It is believed that the anti-aging effect of sirtuins is at least partly dependent on maintaining the regenerative capacity of such cells and of the tissues they support.
However, as mentioned above, there is concern that the anti-aging mechanism(s) of sirtuins may present an inherent danger. For example, they may impair the normal apoptic response that would otherwise eliminate damaged cells, and that by doing so, could encourage overgrowth of abnormal cells and tissues. Further, molecular consequences of sirtuin stimulation, such as down regulation of the tumor suppressor gene p53, may also be a cause for concern about enhancing sirtuin function.
In order to offset the negative effects of sirtuins, “sirtuin-offsetting” agents have been proposed. As used herein, a “sirtuin-offsetting” agent (or “sirtuin-offsetter”) is one that at least partly counteracts the effects of sirtuin as it relates to prolonging cell life. Examples of “sirtuin-offsetting” agents will be discussed in greater detail herein. For example, it has been discovered herein that the decreased activity of certain non-sirtuin HDACs may provide a safeguard. In particular, in certain embodiments of the technology described herein, it has been shown that Class 1, 2 and 4 HDACs have a fundamentally different molecular mechanism and different overall biological effects from the sirtuins, which are Class 3 HDACs. These “non-sirtuin” HDACs do not use NAD as a co-substrate. They also differ from the sirtuins in other ways, e.g., in terms of their target proteins and regulation. Anti-aging or life extension effects have not been demonstrated as consequences of stimulation of non-sirtuin HDACs.
It has been found herein that these HDACs can be targeted for anti-inflammatory benefits and to prevent proliferation of genetically damaged or unstable cells. Non-sirtuin HDAC inhibitors (HDACi's) have been found to be effective at inhibiting proliferation and promoting differentiation or apoptosis and may be useful for cancer treatment, because, among other reasons, in cancer cells many important genes are abnormally repressed by extreme levels of histone deacetylation; thus, the genes that would otherwise be controlling proliferation and initiating differentiation or apoptosis may be inactivated by the non-sirtuin HDACs.
Therefore, in certain embodiments, the present technology is directed to compositions that both stimulate Sirt1 activity (the human sirtuin that is the homolog of yeast Sir2) in order to obtain anti-aging benefits, and also simultaneously inhibit sirtuin-offsetting agents such as, e.g., the non-sirtuin HDACs to the extent sufficient to avoid poorly controlled growth of damaged cells that could eventually compromise tissue function. The compounds discussed herein that activate or inhibit sirtuin are those that have the properties of inhibiting or activating one or more proteins of the sirtuin family—including inhibiting or activating the translation of the mRNAs of a gene, the expression of a gene or the protein activity.
In certain embodiments, the compositions herein provide the dual effect of sirtuin stimulation and inhibition of non-sirtuin protein deacetylase activities, which are not necessarily limited to histones as their targets. That is, histones are proteins but the activity in question can be effective with respect to more than just histone functions. Moreover, as used herein, “protein deacetylase” and “protein deacetylation” refer to histone proteins and to all proteins affected by modulation of acetylation state, not necessarily just histones. Examples of proteins that may be useful targets in this regard include the p53, NF-κB and FOXO transcription factors, the PGC1-α transcriptional coactivator, the DNA repair factor ku70, the AMP-activated protein kinase, LKB1, and the cAMP-responsive coactivator TORC2, among others.
Thus, in certain embodiments, the relative amounts of sirtuin (or sirtuin stimulator, also referred to herein as a sirtuin activator) and sirtuin-offsetting agent (also referred to herein as an inhibitor) in a composition according to the present embodiments is balanced in order to optimize the cell maintenance of the patient. As used herein, “cell maintenance” means the balance of prolonging the life of cells without over-prolonging that can lead to proliferation of damaged tissue and harm to the patient.
In certain embodiments, the present technology provides methods for optimizing the preservation of body cells by balancing the Sirt1 activity of a sirtuin composition with the inhibitory activity of an HDACi.
Data have been developed herein that demonstrate, in in vitro studies, the ability to inhibit preferentially the growth of cells that are genetically damaged by exposure to UV light using a formulation that includes activity for inhibiting non-sirtuin HDACs. Cells were subjected to a sub-lethal dose of UV, but by applying a composition in accordance with certain embodiments herein, inhibition of further growth was shown. The effect was to provide time for the cells to repair, and if not, hold them in a quasi-senescent state until they expired.
In various embodiments the compositions herein may comprise any of the following ingredients:
Water, for example, deionized water, in amounts of about 5 to about 50%, about 10 to about 25% or about 15 to about 20%.
An organic solvent, for example, ethanol or a glycol ether (such as phenoxyethanol, ethylene glycol, propylene glycol or diethylene glycol), in amounts of about 10 to about 50%, about 15 to about 40% or about 15 to about 25%; in certain embodiments, two or more organic solvents in a total amount in such ranges;
A nonionic solubilizer or emulsifying agent having an HLB of about 12 to about 16, for example, a hydroxystearate, in amounts of about 10 to about 25% or about 12 to about 20%;
A botanical extract in amounts of about 20 to about 75%, about 25 to about 60% or about 30 to about 55%.
In various embodiments, suitable botanical extracts include extracts from plants (herbs, roots, flowers, fruits, vegetables, seeds, leaves, pollen, nectar); for example, yeast ferment extract, padica pavonica extract, thermus thermophilis ferment extract, camelina sativa seed oil, boswellia serrata extract, olive extract, aribodopsis thaliana extract, acacia dealbata extract, acer saccharinum (sugar maple), acidophilus, acorns, aesculus, agaricus, agave, agrimonia (for example, Agrimonia species such as Agrimonia eupatoria, Agrimonia gryposepala, Agrimonia incise, Agrimonia coreana, Agrimonia microcarpa, Agrimonia nipponica, Agrimonia parviflora, Agrimonia pilosa, Agrimonia procera, Agrimonia pubescens, Agrimonia repens, Agrimonia rostellata or Agrimonia striata), algae, aloe, citrus, brassica, cinnamon, orange, apple, blueberry, cranberry, peach, pear, lemon, lime, pea, seaweed, caffeine, green tea, chamomile, willowbark, mulberry, poppy, and any other type of botanical extract Further examples include, but are not limited to, Glycyrrhiza Glabra, Salix Nigra, Macrocycstis Pyrifera, Pyrus Malus, Saxifraga Sarmentosa, Vilis Vinifera, Morns Nigra, Scutellaria Baicalensis, Anthemis Nobilis, Salvia Sclarea, Rosmarinus Officianalis, Citrus Medica Limonum, Panax Ginseng, and mixtures thereof.
In certain embodiments, the compositions herein may comprise resveratrol in amounts of about 1 to about 10%, about 2 to about 5% or about 2.5 to about 4%.
In certain embodiments, the compositions herein may comprise either a sirtuin itself, or a sirtuin stimulator or activator, for example, resveratrol or derivatives thereof. Other examples of sirtuin stimulators or activators include: polyphenols and other plant polyphenols, including resveratrol, or resveratrol derivatives (e.g., diphenyl resveratrol, dihydroresveratrol), stilbene, flavone, chalcone, FOXO 3, xanthohumol (found in extracts of hops), isoliquiritigenin (found in extracts of liquorice), phloridzin (found in extracts of apple), piceatannol (found in extracts of rhubarb), natural flavonoids such as fisetin (which is found in extracts of strawberries, grapes, apple or tomato) or epsilon-vineferine.
In certain embodiments, the compositions herein may comprise a non-sirtuin (Class 1 or 2) inhibitor, for example, Psammaplin A thiol, butyrate, diallyl disulfide, Santacruzamate A, sulforaphane, sulforaphane-cysteine, trichostatin A, apicidin, apicidin A, apicidin D1, apicidin D2, azumamide A, azumamide B, azumamide C, azumamide D, azumamide E, chlamydocin, 1-alaninechlamydocin, FR235222, AS1387392, Helminthosporium carbonum toxin, trapoxin A, trapoxin B, Largazole, Largazole thiol, Romidepsin, redFK, any of various hydroxamic acids shown to be HDACi's including suberoylanilide hydroxamic acid, spiruchostatin A, spiruchostatin B, spiruchostatin C, bispyridinium dienes, cyclostellettamine A, cyclostellettamine G, dehydro-cyclostellettamine D, dehydro-cyclostellettamine E, Brazilin, Epicocconigrone A, flavone, pomiferin, apigenin or myricetin.
In certain embodiments, the compositions are particularly useful for applying to the skin of a patient. In certain embodiments, two embodiments may be combined, for example, a sirtuin stimulator including resveratrol or a composition that behaves like resveratrol; and non-sirtuin histone decetylase inhibitor (HDACi) as the sirtuin-offsetting agent.
An exemplary formulation in accordance with the embodiments hereof was prepared as followed:
Deionized water about 12 to about 20%. Zemea propanediol about 10 to about 20%. Phenoxyethanol about 0.25 to about 1%. Solutol HS 15 about 10 to about 20%. Resveratrol about 1 to about 10%. Botanical extract about 45 to about 55%.
In this Example, the water and organic solvents were combined while stirring and heading to about 40 to about 80 degrees C. the Solutol HS 15 and resveratrol were combined thereto with continued stirring. All ingredients were mixed until uniform. The botanical extract was then added to the mixture.
Testing was performed to determine the ability of a prototype formulation in accordance with the present technology, to enhance the apoptotic response of human keratinocytes that had been exposed to DNA-damaging UV radiation. The cells were pretreated with 1% prototype composition for one hour. After washing the cells, they were exposed to 20 mJ/cm2 of UVB light. The cells were then treated for another 21 hours with 1% prototype composition. Cell viability was then measured using the Alamar Blue metabolism method, and results were compared to the viability of unirradiated cells treated similarly with the prototype composition.
Results are shown in
A cream formula was prepared as follows:
A lotion formula was prepared as follows:
As can be seen in
The compositions discussed herein may be in any form that can be applied to the body of a patient; for example, to the skin. In certain embodiments, they may be cosmetically or pharmaceutically acceptable forms that can be incorporated into lotions, creams, sprays, gels, serums, liquids, suspensions or the like. Encapsulation technologies such as liposomes, micellar constructs and the like are also contemplated.
The present application is a continuation-in-part application of U.S. application Ser. No. 14/518,860, filed on Oct. 20, 2014, entitled “COMPOSITIONS AND METHODS COMPRISING SIRTUIN STIMULATORS AND NON-SIRTUIN PROTEIN DEACETYLASE INHIBITORS”, which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| 61951256 | Mar 2014 | US | |
| 61892867 | Oct 2013 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 14518860 | Oct 2014 | US |
| Child | 15266446 | US |
