Senolytic and Anti-aging compositions including camellia nitidissima chi, and methods

Abstract

Described herein are methods and compositions for use in treating, preventing or reversing aging or age-related diseases, disorders or conditions. Also, described herein are methods and compositions for selective killing of senescent cells that are associated with numerous pathologies and diseases, including age-related pathologies, diseases and conditions. As disclosed herein are methods where senescent cell associated diseases and disorders may be treated or prevented (i.e., likelihood of occurrence or development is reduced) by administering the described compositions.

Description

SENOLYTIC AND ANTI-AGING COMPOSITIONS INCLUDING CAMELLIA NITIDISSIMA CHI, AND METHODS BACKGROUND

[0001] Aging in an organism is accompanied by an accumulation of changes over time. Human beings dream of extending their life span.

[0002] Senescent cells accumulate in tissues and organs of individuals as they age and are found at sites of age-related pathologies. Senescent cells display senescence- associated expression of beta galactosidase activity. Beta galactosidase is a hydrolase enzyme that cleaves beta-linked terminal galactosyl residues from gangliosides, glycoproteins, and glycosaminoglycans. It is an essential marker of senescent cells. Its staining is widely used in aging-related detection both in vitro and in vivo.

[0003] Senescent cells are believed important to inhibiting proliferation of dysfunctional or damaged cells and particularly to constraining development of malignancy (see, e.g., Campisi, Curr. Opin. Genet. Dev.21:107-12 (2011); Campisi, Trends Cell Biol.11:S27-31 (2001); Prieur et al., Curr. Opin. Cell Biol.20:150-55 (2008)). Nevertheless, the presence of senescent cells in an individual may contribute to aging and aging-related dysfunction (see, e.g., Campisi, Cell 120:513-22 (2005)). Given that senescent cells have been causally implicated in certain aspects of age-related decline in health and may contribute to certain diseases, and are also induced as a result of necessary life-preserving chemotherapeutic and radiation treatments, the presence of senescent cells may have deleterious effects to millions of patients worldwide. However, identifying and developing treatments of such diseases and conditions by selective elimination of senescent cells has been an arduous undertaking. The present disclosure addresses these needs and offers related advantages.

[0004] Proof-of-concept studies have led to the identification of compounds that can selectively eliminate senescent cells (so-called “senolytics”).

[0005] Several senolytic compounds have been identified to date, including dasatinib and quercetin (Zhu et al., 2015), piperlongumine (Wang et al., 2016) and BCL2-family inhibitors such as ABT263 and ABT737 (Chang et al., 2016; Yosef et al., 2016; Zhu et al., 2016). Currently BCL2 family inhibitors are the most promising senolytics, having been shown to kill a range of senescent cells in vivo, with reproducible effects intransgenic mouse models (Chang et al., 2016; Childs et al., 2016; Yosef et al., 2016; Zhu et al., 2016). BCL2 inhibitors were initially developed as therapies for lymphoma. ABT- 737 is a small molecule inhibitor of BCL-2, BCL-XL and BCL-w but has low solubility and oral bioavailability. ABT-263 inhibits the same molecules and is better suited for use in vivo but causes significant thrombocytopenia as a side-effect.

[0006] Due to the significant side-effects exhibited by senolytic compounds identified to date, there is a great need to identify further compounds, and classes of compounds, with senolytic properties.

[0007] Camellia nitidissima Chi or CNC is common traditional ethnic medicine in Guangxi Province, South China, and North Vietnam (Yang, Q. H., et al. (2008). Seed biology and germination ecophysiology of Camellia nitidissima. Forest Ecology and Management, 255(1)). Camellia nitidissima Chi belongs to the Camellia genus (Theaceae family). As a rare species in the world, Camellia nitidissima Chi often referred to as the “panda of the plant kingdom” in China (Song LX, et al., “Polyphenolic antioxidant profiles of yellow camellia.” Food Chem.2011; 129(2):351–7).

[0008] Camellia nitidissima Chi has previously been found to help with clearing of heat and toxic materials and diuretic detumescence. In recent years, a great number of investigations have shown that this plant contains a rich source of bioactive constituents, including flavonoids, polysaccharides, saponins, and other components. Pharmacological studies have demonstrated that Camellia nitidissima Chi possesses anti-tumor, anti- inflammatory, antioxidant, hypolipidemic effect, hypoglycemic, anti-obesity, hypotensive, and anti-anaphylaxis properties (Huang, Y., et al., “Effects of the extracts from Camellia nitidissima leaves on blood lipids,” Lishizhen Medicine and Materia Medica Research, 2009–04(20), 776–777 (2009); Li, M. H., et al., “Nuclear magnetic resonance-based metabolomics approach to evaluate the prevention effect of Camellia nitidissima Chi on colitis-associated carcinogenesis,” Frontiers in Pharmacology, 8, 447– 464 (2017); Yang, R., et al., “Antioxidant capacity of phenolics in Camellia nitidissima Chi flowers and their identification by HPLC Triple TOF MS / MS,” PLoS One, 13, e0195508–e0195520 (2018); and Yang, R., et al., “The inhibition of advanced glycation end-products by five fractions and three main flavonoids from Camellia nitidissima chi flowers,” Journal of Food & Drug Analysis, 26(1), 252–259 (2018)).SUMMARY

[0009] An embodiment relates to a composition for use in treating, preventing or reversing aging or age-related disease, disorder or condition, comprising an alcohol or a water extract of leaves of Camellia nitidissima Chi and a pharmaceutically acceptable excipient. The age-related disease, disorder, or condition may be skin conditions, skin wound healing, renal disease, renal failure, frailty, hearing loss, muscle fatigue, muscle loss, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia. The composition may be formulated for oral administration or for a topical administration. The composition may be in a form of an aerosol, a cream, a emulsion, a solid, a liquid, a dispersion, a foam, a gel, a lotion, a mousse, an ointment, a powder, a patch, a pomade, a solution, a pump a spray, a stick, am oil, a towelette, or combinations thereof. In the composition, the alcohol or water extract of Camellia nitidissima Chi may be present in an amount of between about 0.1 to about 26.67 mg / kg (60 kg per person). The composition may further comprise an extract of Lycium ruthenicum.

[0010] Another embodiment relates to a composition for use in selectively killing one or more senescent cells, comprising an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically acceptable excipient.

[0011] Yet another embodiment relates to a composition for use in treating or preventing a senescence-associated disease or disorder, comprising an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically acceptable excipient. The senescence-associated disease or disorder may be a cardiovascular disease or disorder, a metabolic disease, an inflammatory disease or disorder, a pulmonary disease or disorder, a neurological disease or disorder, a renal disorder or disease, or a dermatological disorder or disease. The senescence-associated disease or disorder is selected from: (i) an inflammatory or autoimmune disease or disorder selected from osteoarthritis, osteoporosis, oral mucositis, inflammatory bowel disease, kyphosis and herniated intervertebral disc; (ii) a neurological disease or disorder selected from Alzheimer's disease, Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment, macular degeneration and motor neuron dysfunction; (iii) a metabolic disease selected from diabetes, diabetic ulcer, metabolic syndrome and obesity; (iv) a pulmonary disease selected from pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cysticfibrosis, emphysema, bronchiectasis and age-related loss of pulmonary function; (v) an eye disease or disorder selected from macular degeneration, glaucoma, cataracts, presbyopia and vision loss; (vi) an age-related disorder selected from renal disease, renal failure, frailty, hearing loss, muscle fatigue, skin conditions, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia; (vii) a dermatological disease or disorder selected from eczema, psoriasis, hyperpigmentation, nevi, rashes, atopic dermatitis, urticaria, diseases and disorders related to photosensitivity or photoaging, rhytides, pruritis, dysesthesia, eczematous eruptions, eosinophilic dermatosis, reactive neutrophilic dermatosis, pemphigus, pemphigoid, immunobullous dermatosis, fibrohistocytic proliferations of skin, cutaneous lymphomas and cutaneous lupus; and (viii) a cardiovascular disease selected from atherosclerosis, angina, arrhythmia, cardiomyopathy, congestive heart failure, coronary artery disease, carotid artery disease, endocarditis, coronary thrombosis, myocardial infarction, hypertension, aortic aneurysm, cardiac diastolic dysfunction, hypercholesterolemia, hyperlipidemia, mitral valve prolapse, peripheral vascular disease, cardiac stress resistance, cardiac fibrosis, brain aneurysm and stroke.

[0012] Yet another embodiment relates to a nutritional supplement compositing an extract of Camellia nitidissima Chi and a neutraceutically acceptable excipient. In the nutritional supplement, the extract may be an alcohol extract or a water extract of the leaves of Camellia nitidissima Chi. In the nutritional supplement, the extract may be an alcohol extract of the flowers of Camellia nitidissima Chi. The nutritional supplement may further comprise an extract of Lycium ruthenicum.

[0013] Yet another embodiment relates to a method for treating, preventing or reversing aging or age-related disease, disorder, or condition in a subject, comprising administering to the subject a composition comprising a therapeutically effective amount of an alcohol or a water extract of the leaves of Camellia nitidissima Chi and a pharmaceutically acceptable excipient. The composition may further comprise an extract of Lycium ruthenicum. In the composition, the age-related disease, disorder, or condition may be skin conditions, skin wound healing, renal disease, renal failure, frailty, hearing loss, muscle fatigue, muscle loss, liver fibrosis, pancreatic fibrosis, oral submucosafibrosis and sarcopenia. The composition may be formulated for oral administration, or may be formulated for a topical administration.

[0014] Yet another embodiment relates to a method for selectively killing one or more senescent cells in a subject, comprising administering to the subject a therapeutically effective amount of an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically, acceptable excipient. In the method, the senescence-associated disease or disorder may be a cardiovascular disease or disorder, a metabolic disease, an inflammatory disease or disorder, a pulmonary disease or disorder, a neurological disease or disorder, a renal disorder or disease, or a dermatological disorder or disease.

[0015] Yet further embodiment relates to a composition for use in treating, preventing or reversing aging or age-related disease, disorder or condition, comprising an alcohol or a water extract of leaves of Camellia nitidissima Chi; an extract of Lycium ruthenicum; and a pharmaceutically acceptable excipient. The composition may be for use as a dietary supplement, for use as a cosmetic product, and / or for use in anti-aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is a graph of cell viability of leaves water extract on Hs27 human fibroblasts. Cells were treated with 6 concentrations (15.63, 31.25, 62.50, 125.00, 250.00 and 500,00 µg / ml) for 72 h. Cell proliferation was determined by CCK8 assay. Values are represented as the means ± SD.15.63 µg / ml was used as max concentration for the next testing since the cell viability is higher than 80%.

[0017] Figure 2 depicts a graph of Cell viability of leaves alcoholic extract on Hs27 human fibroblasts. Cells were treated with 6 concentrations (15.63, 31.25, 62.50, 125.00, 250.00 and 500,00 µg / ml) for 72 h. Cell proliferation was determined by CCK8 assay. Values are represented as the means ± SD. Values are represented as the means ± SD. 15.63 µg / ml was used as max concentration for the next testing since the cell viability is higher than 80%.

[0018] Figure 3 shows anti-aging function of either water or alcoholic extract from leaves. (A.) D-galactose was used to generate a model of accelerated aging on Hs27. Cells were treated either water or alcoholic extracts from leaves. Aging cell were stained by SA-^-Gal positivity after 72 h treatment (blue color). (B.) Aging cell ratio aftertreatment of different concentrations of water extract from leaves. Values are represented as the means ± SE. ** indicates p < 0.001 compared with Model group. (C.) Aging cell ratio after treatment of different concentrations of alcoholic extract from leaves. Values are represented as the means ± SE. ** indicates p < 0.001 compared with Model group.

[0019] Figure 4 depicts senolytic potential of alcoholic extract from flowers. (A.) Mitomycin C was used to generate a model of accelerated aging on WI-38. Aging cell were stained by SA-^-Gal positivity (black dots). either water or alcoholic extracts from leaves. (B.) Both aging and normal cells were treated with alcoholic extract from flowers for 7-day treatment. Cell viabilities were detected after treatments ratio after treatment. DETAILED DESCRIPTION OF THE DRAWINGS AND THE PRESENTLY PREFERRED EMBODIMENTS

[0020] The present invention will now be described more fully herein after. For the purposes of the following detailed description, it is to be understood that the invention may assume various alternative variations and step sequences, except where expressly specified to the contrary. Thus, before describing the present invention in detail, it is to be understood that this invention is not limited to particularly exemplified embodiments that may of course, vary.

[0001] Unless otherwise defined, all terms technical and scientific used in this specification generally have their ordinary meanings in the art, within the context of the invention, and in the specific context where each term is used. Certain terms that are used to describe the invention are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the invention.

[0002] As used herein, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. The term “and / or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0003] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that otherembodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.

[0004] As used herein, “around,” “about” or “approximately” shall generally mean within 20 percent, preferably within 10 percent, and more preferably within 5 percent of a given value or range. Numerical quantities given herein are approximate; meaning that the terms “around,” “about” or “approximately” can be inferred if not expressly stated. When the term “about” is used in describing a value or an endpoint of a range, the disclosure should be understood to include both the specific value and end-point referred to.

[0005] As used herein, the terms “comprising,” “including,” “having,” “containing,” “involving,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to.

[0006] The term “composition” is used herein to describe a formulation that includes a pure compound, composition, extract, extract mixture, component of the extract, and / or active agent or ingredient, or a combination thereof of Camellia nitidissima Chi. The composition is suitable for, e.g., oral ingestion by the subject (e.g., in a form of a nutritional, neutraceutical, or pharmaceutical product, such as a supplement) or topical application (e.g., in a form of a pharmaceutical or cosmetic product). Exemplary compositions that include a pure compound, composition, extract, extract mixture, component of the extract, and / or active agent or ingredient, or a combination thereof of Camellia nitidissima Chi include, but are not limited to: sprays (e.g., aerosols), powders, chewing gum, ingestible solids, gels, supplement products, aqueous beverages, dry powder (e.g., a powder that can be directly consumed or that can be reconstituted with liquid to provide a beverage), nutritional bars, lozenges, pills, tablets, capsules, wafers, pastes, and the like. Other compositions are described herein.

[0007] The terms “extract” or “botanical extract” refer to a solid, viscid, or liquid substance or preparation obtained from the entire Camellia nitidissima Chi plant (i.e., plant material) or a part thereof (e.g., leaves, flowers, etc. of Camellia nitidissima Chi plant). It is a material which dissolves in a solvent after contacting a plant substance with a suitable solvent. Preferably, the plant substance is contacted for a sufficiently long period of time and at a suitable temperature so that substantially all of the soluble materialis removed by the solvent (e.g., water, alcohol (e.g., ethanol)). The solvent together with the dissolved soluble plant material is referred to as the “liquid extract.” The term “extract” refers to the soluble plant material remaining after removal of the solvent. The solvent can be removed by any suitable means, including evaporation, lyophilization, spray drying and the like.

[0008] The term “extract” is intended to include not only a crude extract produced from Camellia nitidissima Chi, and by use of a solvent selected from among water, lower alcohols of 1 to 4 carbon atoms, such as methanol, ethanol, butanol, etc., ethylene, acetone, hexane, ether, chloroform, ethylacetate, butylacetate, dichloromethane, N,N- dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol and a combination thereof, but also a fraction of the crude extract in such a solvent. So long as it assures the extraction and preservation of the active ingredient(s), any extraction method may be employed.

[0009] The terms “cosmetically acceptable” and “pharmaceutically acceptable” mean those drugs, medicaments, supplements, ointments, extracts or inert ingredients, which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, incompatibility, instability, irritation, and the like, commensurate with a reasonable benefit / risk ratio.

[0010] The terms “administering” and “applying” can use interchangeably and are defined as providing a composition to a subject via a route known in the art, including but not limited to topical, oral, intravenous, intraarterial, parenteral, buccal, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal routes of administration. In preferred embodiments, topical routes of administering (i.e., “topically applying”) a composition are suitable.

[0011] The term “prevent” in the context of preventing aging or aging-related diseases, disorders, or conditions in a subject means to keep the aging process and / or the aging-related diseases, disorders, or conditions from happening or existing.

[0012] As used herein, the term “subject” or “individual” includes mammals to which a composition may be administered or applied. Non-limiting examples of mammals include humans, non-human primates, rodents (including transgenic and non-transgenic mice) or the like. In some embodiments, the subject is a mammal, and in some embodiments, the subject is human.

[0013] As used herein, the term “senolytic” refers to a compound, ingredient, extract, or a composition that selectively (preferentially or to a greater degree) destroys, kills, removes, or facilitates selective destruction of senescent cells, i.e. the compound, ingredient, extract or the composition destroys or kills a senescent cell in a biologically, clinically, and / or statistically significant manner compared with its capability to destroy or kill a non-senescent cell. The senolytic compound is used in an amount and for a time sufficient to selectively kill established senescent cells, but which is insufficient to kill non-senescent cells in a clinically significant or biologically significant manner. In the context of this application, an ingredient or an extract of Camellia nitidissima Chi plant or part thereof may be considered senolytic.

[0014] As used herein, the terms “sufficient,” “effective” or “therapeutically effective” with reference to the amount or time refer to an amount effective at dosages and for periods of time sufficient, respectively, to achieve a desired result. For example, the “amount sufficient,” “effective amount” or “therapeutically effective amount” refer to that amount of a pure compound, composition, extract, botanical extract, extract mixture, botanical extract mixture, component of the extract, and / or active agent or ingredient, or a combination thereof which, when administered to a subject (e.g., mammal, such as a human), is sufficient to effect reversal, prevention, treatment, and or cure of, e.g., aging and / or aging-associated disease, disorder or condition in the subject.

[0015] Aging is a risk factor for most chronic diseases, disabilities, and declining health. Senescent cells, which are cells in replicative arrest, accumulate as an individual ages and may contribute partially or significantly to cell and tissue deterioration that underlies aging and age related diseases. Cells may also become senescent after exposure to an environmental, chemical, or biological insult or as a result of a disease.

[0016] Described herein are methods and compositions for use in treating, preventing or reversing aging, e.g., signs of aging, such as wrinkles, or treating, preventing age- related diseases, disorders or conditions. Also, described herein are methods and compositions for selective killing of senescent cells that are associated with numerous pathologies and diseases, including age-related pathologies, diseases and conditions.Also, disclosed herein are methods where senescent cell associated diseases and disorders may be treated or prevented (i.e., likelihood of occurrence or development is reduced) by administering the described compositions.

[0017] The terms “pathological” or “pathology” mean that the effect is caused by or involving disease.

[0018] The term “non-pathological” means that the effect is caused by or involving something (e.g., normal aging process), other than disease. For example, non-pathological condition may be age-related condition, such as skin aging, lines, fine lines, wrinkles, crow's feet, dark eye circles, age spots, dark spots, etc.

[0019] The terms “reduce,” “reducing,” “inhibit” or “inhibiting” refer to a decrease or reduction in an effect, in the presence of a plant ingredient or plant extract of Camellia nitidissima Chi when compared to the same effect in the absence of a plant ingredient or plant extract of Camellia nitidissima Chi, such as in a control sample. The degree of decrease or inhibition will vary with the nature and quantity of a plant ingredient or plant extract of Camellia nitidissima Chi present, but will be evident, e.g., as a detectable decrease; desirably a degree of decrease greater than 1%, 2,%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 50%, 75%, 90%, 95% or 99% as compared to a sample (without Camellia nitidissima Chi).

[0020] Described herein are compositions comprising Camellia nitidissima Chi or “CNC” as the active ingredient or active extract for use in anti-aging. In certain embodiments, the active ingredient or an extract of Camellia nitidissima Chi is the only active in a composition.

[0021] Some aspects relate to compositions for treating, preventing or reversing aging (e.g., signs of aging, such as wrinkles) or age-related diseases, disorders or conditions, and methods relating thereto.

[0022] Also, the described compositions may be for use in selectively killing one or more senescent cells.

[0023] Further aspects relate to compositions for use in treating or preventing senescence-associated diseases, disorders or conditions, and methods relating thereto.

[0024] The compositions described herein may be considered senolytic.

[0025] The composition described herein may be considered anti-aging.

[0026] In certain embodiments, the senolytic compositions described herein can alter at least one signaling pathway in a manner that induces (initiates, stimulates, triggers, activates, promotes) and results in death of the senescent cell. A senolytic composition may alter one or more signaling pathways in a senescent cell by interacting with one or more target proteins, which results in removing or reducing suppression of a cell death pathway, such as an apoptotic pathway. For example, contacting or exposing a senescent cell to a senolytic composition may restore the cell's mechanisms and pathways for initiating apoptosis. In one embodiment, the composition induces apoptosis.

[0027] Camellia nitidissima Chi or CNC is a plant of Camellia characterized with its golden-yellow flowers which is mainly distributed in a narrow region of Southern China and Northern Vietnam. CNC is called as “the Queen of Camellia” (He et al., 2018) and listed as a medicinal plant in Guangxi Ethnomedicines Compendium (He et al., 2019). CNC contains various active ingredients including tea polysaccharides, polyphenols, flavonoids, tea saponins and volatile components, etc. (He et al., 2018). Recent studies indicate that CNC has the bioactivities of antioxidant, anti-glycation, anticancer and neuroprotection (An et al., 2020, He et al., 2019, Wang et al., 2016, Yang et al., 2018).

[0028] Camellia nitidissima Chi has been found to be rich in tea polysaccharides, tea polyphenols, total saponin, total flavones, tea pigments, caffeine, protein, vitamin B1, B2, vitamin C, vitamin E, folic acid, fatty acid, B-carotene, amino acids such as theanine, threonine, and macroelements, such as the trace element such as the natural organic germanium, selenium, molybdenum, zinc, vanadium, potassium, calcium, and magnesium.

[0029] The extracts of Camellia nitidissima Chi can be prepared by the following methods:

[0030] For example, in certain embodiments, dried leaves of Camellia nitidissima Chi (50 g) may be refluxed in 500 mL of water solution at 95 ºC in a water bath for 1.5 h and then in 350 mL of fresh solution for a further 1 h. The water extracts can be combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure.

[0031] For alcoholic extracts, dried leaves of Camellia nitidissima Chi (50 g) can be refluxed in 500 mL of 70% (v / v) ethanol solution at 95 ºC in a water bath for 1.5 h and then in 350 mL of fresh solution for a further 1 h. The ethanol extracts can be combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure. Besides, to make alcoholic extract of flower, dried flowers of Camellia nitidissima Chi (50 g) can be refluxed in 600 mL of 70% (v / v) ethanol solution at 95 ºC in a water bath for 1.5 h and then in 400 mL of fresh solution for a further 1 h. The ethanol extracts can be combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure. These three extract-powders can be obtained by drying the concentrated solution using a freeze dryer and stored in a refrigerator at -20°C until use.

[0032] The raw leaves and flowers of Camellia nitidissima Chi were obtained from Fangchenggang, a city in the south of Guangxi Zhuang Autonomous Region, People’s Republic of China.

[0033] Alternatively, leaves and flowers of Camellia nitidissima Chi are legally available as food in China, and there are many suppliers available.

[0034] The yield of leaves of Camellia nitidissima Chi is the range of from about 1% to 40%; preferably about 8%.

[0035] The yield of flowers of Camellia nitidissima Chi is the range of from about range of 1% to 70%; and preferably about 32%.

[0036] The results of the composition identification showed that ellagic acid derivatives (e.g., okicamelliaside), flavonoids (e.g., vicenin-1, vicenin-2), saponins and proanthocyanidins are the main components of Camellia nitidissima Chi.

[0037] In one example, a botanical extract useful in the compositions described herein might be obtained using a deionized water extraction technique.

[0038] In another example, a botanical extract useful in the compositions described herein might be obtained using an alcohol, e.g., ethanol, extractions methods.

[0039] Those of skill in the art will appreciate that there are many other extraction processes, both known in the art and described in various patents and publications that can be used to obtain the extracts to be used in practicing the present invention.

[0040] Certain embodiments relate to a composition for use in treating, preventing or reversing aging or age-associated diseases, disorders or conditions. The compositioncomprises an ingredient or an extract of Camellia nitidissima Chi and a pharmaceutically, neutraceutically, or cosmetically acceptable excipient. In certain embodiments, the extract is an alcohol extract of the leaves of Camellia nitidissima Chi. In certain other embodiments, the extract is a water extract of the leaves of Camellia nitidissima Chi.

[0041] Certain other embodiments relate to a composition for use in selectively killing one or more senescent cells. The composition comprises an ingredient or an extract of Camellia nitidissima Chi and a pharmaceutically, neutraceutically, or cosmetically acceptable excipient. In certain embodiments, the extract is an alcohol extract of the flower of Camellia nitidissima Chi.

[0042] In preferred embodiments, the described compositions selectively kill senescent cells over non-senescent cells. Preferably, the described compositions exhibit a selectivity for killing senescent cells over non-senescent cells of at least 1.5 fold, more preferably, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20- fold, at least 25-fold, at least 50-fold, or at least 100-fold.

[0043] Certain further embodiments relate to a composition for use in treating or preventing a senescence-associated disease or disorder. The composition comprises an ingredient or an extract of Camellia nitidissima Chi and a pharmaceutically, neutraceutically, or cosmetically acceptable excipient. In certain embodiments, the extract is an alcohol extract of the flower of Camellia nitidissima Chi.

[0044] Another aspect relates to compositions as defined herein for use as a senolytic. The senolytic activity of the described compositions is demonstrated with reference to the accompanying examples, and is described in more detail below. For example, Figure 4 demonstrates the Senolytic potential of alcoholic extract from flowers of Camellia nitidissima Chi. Both aging and normal cells were treated with alcoholic extract from flowers of Camellia nitidissima Chi for 7-day treatment. Cell viabilities were detected after treatments ratio after treatment.

[0045] Therapeutic Applications

[0046] The described compositions may be for use in treating, preventing or reversing aging, e.g., signs of aging, such as wrinkles, and / or treating, preventing age-related diseases, disorders or conditions, and methods relating thereto.

[0047] In certain embodiments, the described compositions may be for use in anti- aging.

[0048] Also, in certain other embodiments, the described compositions may be for use in selectively killing one or more senescent cells.

[0049] Further aspects relate to compositions for use in treating or preventing senescence-associated diseases, disorders or conditions, and methods relating thereto.

[0050] In another aspect, the compositions described herein can be used to treat a wide range of different age-related diseases, disorders or conditions.

[0051] In yet further aspect, the compositions described herein can be used to treat a wide range of different senescence-associated diseases, disorders or conditions.

[0052] For example, in one embodiment, the described composition delays tumorigenesis. In another embodiment, the described composition attenuates cataract formation. In another embodiment, the described composition attenuates atherosclerosis. In another embodiment, the described composition attenuates the age-related deterioration of kidney, fat and heart amongst other organs.

[0053] The presence of senescent cells can also be determined by detecting other senescent cell-associated molecules, such as cytokines, chemokines, expressed and secreted growth factors, proteases, cell-related metabolites, reactive oxygen species, and other molecules that stimulate inflammation and / or other biological effects. Examples of senescent cell-associated molecules include those that comprise the senescence-associated secretory phenotype (SASP), senescent-messaging secretome, and DNA damage secretory program (DDSP).

[0054] As used herein, senescence-associated disorders, diseases or conditions include disorders, diseases or conditions associated with, or caused by cellular senescence, including age-related diseases, disorders or conditions. A senescence- associated disease, disorder or condition may also be called a senescent cell-associated disease, disorder, or condition. A prominent feature of aging is a gradual loss of function, or degeneration that occurs at the molecular, cellular, tissue, and organismal levels. Age- related degeneration gives rise to well-recognized pathologies such as sarcopenia, atherosclerosis and heart failure, osteoporosis, pulmonary insufficiency, renal failure,neurodegeneration (including macular degeneration, Alzheimer's disease, and Parkinson's disease), hair and skin changes, and many others.

[0055] Senescence-associated diseases and disorders include, but are not limited to, cardiovascular diseases and disorders, proliferative disorders such as cancer, inflammatory diseases and disorders, autoimmune diseases and disorders, pulmonary diseases and disorders, eye diseases and disorders, metabolic diseases and disorders, neurological diseases and disorders (e.g., neurodegenerative diseases and disorders); age- related diseases and disorders induced by senescence; skin conditions; age-related diseases; dermatological diseases and disorders; and transplant related diseases and disorders.

[0056] Preferably, the subject is a mammal, more preferably a human.

[0057] The term “method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0058] Herein, the term “treating” includes abrogating, substantially inhibiting, slowing or reversing the progression of a disease, disorder or condition, substantially ameliorating clinical symptoms of a disease, disorder or condition, or substantially preventing the appearance of clinical symptoms of a disease, disorder, or condition. Treatment also includes situations where a pathological condition, or at least symptoms associated therewith, are completely inhibited, e.g., prevented from happening, or stopped, e.g., terminated, such that the adult mammal no longer suffers from the impairment, or at least the symptoms that characterize the impairment. In some instances, “treatment”, “treating” and the like refer to obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. “Treatment” may be any treatment of a condition and / or disease in a mammal, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e.,arresting its development; or (c) relieving the disease, i.e., causing regression of the disease. Treatment may result in a variety of different physical manifestations, e.g., modulation in gene expression, increased neurogenesis, rejuvenation of tissue or organs, etc. Treatment of ongoing disease, where the treatment stabilizes or reduces the undesirable clinical symptoms of the patient, occurs in some embodiments. Such treatment may be performed prior to complete loss of function in the affected tissues. The subject therapy may be administered during the symptomatic stage of the condition or disease, and in some cases after the symptomatic stage of the condition or disease.

[0059] In some instances where the aging-related impairment is aging-associated skin aging, treatment by methods of the present disclosure slows, or reduces, the progression of aging-associated skin aging. In other words, skin aging in the individual progresses more slowly, if at all, following treatment by the disclosed methods than prior to or in the absence of treatment by the disclosed methods.

[0060] Herein, the term “preventing” refers to a method for barring an organism from acquiring a disorder or disease in the first place.

[0061] Herein, the term “reversing” refers to a method for regression of the disease in the subject.

[0062] The term “therapeutically effective amount” refers to that amount of the agent being administered which will relieve to some extent one or more of the symptoms of the disease or disorder being treated.

[0063] For an ingredient, an extract of Camellia nitidissima Chi or a composition thereof used herein, a therapeutically effective amount, also referred to herein as a therapeutically effective dose, can be estimated initially from cell culture assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC50 or the IC100 as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Initial dosages can also be estimated from in vivo data. Using these initial guidelines one of ordinary skill in the art could determine an effective dosage in humans.

[0064] Moreover, toxicity and therapeutic efficacy of the ingredient, an extract of Camellia nitidissima Chi or a composition thereof described herein can be determined bystandard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD50and the ED50. The dose ratio between toxic and therapeutic effect is the therapeutic index and can be expressed as the ratio between LD50 and ED50. The data obtained from these cell cultures assays and animal studies can be used in formulating a dosage range that is not toxic for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual and / or individual’s physician in view of the patient's condition. (see, e.g., Fingl et al, 1975, In: The Pharmacological Basis of Therapeutics, chapter 1, page 1).

[0065] Dosage amount and interval may be adjusted individually to provide plasma levels of the active compound which are sufficient to maintain therapeutic effect. Preferably, therapeutically effective serum levels will be achieved by administering multiple doses each day. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration. One skilled in the art will be able to optimize therapeutically effective local dosages without undue experimentation.

[0066] Proliferative Disorders

[0067] In certain embodiments, the described compositions can be used in the treatment of a proliferative disorder such as cancer or leukemia.

[0068] Non-limiting examples of cancers include: adenocarcinoma, adrenal gland tumor, ameloblastoma, anaplastic, anaplastic carcinoma of the thyroid, angiofibroma, angioma, angiosarcoma, apudoma, argentaffmoma, arrhenoblastoma, ascites tumor cell, ascitic tumor, astroblastoma, astrocytoma, ataxia-telangiectasia, atrial myxoma, a basal cell carcinoma cell, bone cancer, brainstem glioma, brain tumor, breast cancer, Burkitt's lymphoma, cerebellar astrocytoma, cervical cancer, cherry angioma, cholangiocarcinoma, cholangioma, chondroblastoma, chondroma, chondrosarcoma, chorioblastoma, choriocarcinoma, colon cancer, common acute lymphoblastic leukemia, craniopharyngioma, cystocarcinoma, cystofbroma, cystoma, ductal carcinoma in situ, ductal papilloma, dysgerminoma, encephaloma, endometrial carcinoma, endothelioma,ependymoma, epithelioma, erythroleukemia, Ewing's sarcoma, extra nodal lymphoma, feline sarcoma, fibro adenoma, fibro sarcoma, follicular cancer of the thyroid, ganglioglioma, gastrinoma cell, glioblastoma multiform, glioma, gonadoblastoma, haemangioblastoma, haemangioendothelioblastoma, haemangioendothelioma, haemangiopericytoma, haematolymphangioma, haemocytoblastoma, haemocytoma, hairy cell leukemia, hamartoma, hepatocarcinoma, hepatocellular carcinoma, hepatoma, histoma, Hodgkin's disease, hypernephroma, infiltrating cancer, infiltrating ductal cell carcinoma, insulinoma, juvenile angioforoma, Kaposi sarcoma, kidney tumor, large cell lymphoma, leukemia, a leukemia, acute leukemia, lipoma, liver cancer, liver metastases, Lucke carcinoma, lymphadenoma, lymphangioma, lymphocytic leukemia, lymphocytic lymphoma, lymphoeytoma, lymphoedema, lymphoma, lung cancer, malignant mesothelioma, malignant teratoma, mastocytoma, medulloblastome, melanoma, meningioma, mesothelioma, Morton's neuroma, multiple myeloma, myeloblastoma, myeloid leukemia, myelolipoma, myeloma, myoblastoma, myxoma, nasopharyngeal carcinoma, neoplastic, nephroblastoma, neuroblastoma, neurofibroma, neurofibromatosis, neuroglioma, neuroma, non-Hodgkin's lymphoma, oligodendroglioma, optic glioma, osteochondroma, osteogenic sarcoma, osteosarcoma, ovarian cancer, Paget's disease of the nipple, pancoast tumor, pancreatic cancer, phaeochromocytoma, pheoehromocytoma, plasmacytoma, primary brain tumor, progonoma, prolactinoma, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, rhabdo sarcoma, a solid tumor, sarcoma, a secondary tumor, seminoma, skin cancer, small cell carcinoma, squamous cell carcinoma, strawberry haemangioma, T-cell lymphoma, teratoma, testicular cancer, thymoma, trophoblastic tumor, Wilm's tumor.

[0069] In certain embodiments, the described compositions can be used in conjunction with other treatments for cancer that induce senescence, such as irradiation or chemotherapy (for example, treatment with Palbociclib, ribociclib or abemaciclib, or other chemotherapeutic agents). Thus, by treating at the same time, or after the other treatment, the agent can:

[0070] eliminate cancer cells that have been pushed to senescence; and / or

[0071] eliminate or reduce certain side effects produced by senescent cells such as inflammation, promotion of cancer growth, promotion of metastasis and other side effects of chemotherapy or radiotherapy; and / or

[0072] reduce or eliminate precancerous lesions; and / or

[0073] eliminate or reduce cells that undergo senescence by treatment with CDK4 / 6 inhibitors.

[0074] In certain embodiments, the described compositions can reduce or eliminate precancerous (or pre-neoplastic) lesions. Senescent cells exist in premalignant tumors, but not in malignant ones. In this regard, it is understood that a substantial number of cells in premalignant tumors undergo oncogene-induced senescence, but that cells in malignant tumors are unable to do this owing to the loss of oncogene-induced senescence effectors such as p16INK4aor p53 (Collado et al; Nature, Vol 436, August 2005, page 642). Thus, one embodiment of the invention relates to an agent as described herein for use in reducing or eliminating precancerous (or pre-neoplastic) lesions.

[0075] In certain embodiments, the described compositions can eliminate or reduce chemotherapy-induced senescence, for example, before, during or after treatment with a chemotherapeutic agent. Chemotherapeutic agents include, but are not limited to, anthracyclines, doxorubicin, etoposide, daunorubicin, taxols, paclitaxel, gemcitabine, pomalidomide, and lenalidomide. Thus, in certain embodiments, the described compositions can be used in combination treatment with a chemotherapeutic agent, where the composition is administered separately, sequentially or concomitantly with the chemotherapeutic agent. One embodiment of the invention relates to the described composition for use in eliminating or reducing chemotherapy-induced senescence.

[0076] In certain embodiments, the described compositions can eliminate or reduce senescence induced by treatment with a CDK inhibitor, for example, a CDK4 or CDK6 inhibitor. Thus, in one preferred embodiment, the agent can be used in combination treatment with a CDK4 or CDK6 inhibitor, where the agent is administered separately, sequentially or concomitantly with the CDK4 or CDK6 inhibitor.

[0077] In certain embodiments, the described compositions can eliminate or reduce Palbociclib-induced senescence. In the context of eliminating or reducing Palbociclib-induced senescence, administration of the agent can potentially preventcancer remission as cells reenter the cell cycle (see Cadoo et al; Breast Cancer: Targets and Therapy 2014; 6, 123-133).

[0078] In certain embodiments, the described compositions can eliminate cancer cells that have been pushed to senescence. In certain embodiments, the described compositions delay tumorigenesis.

[0079] In certain embodiments, the described compositions can eliminate or reduce certain side effects produced by senescent cells such as inflammation, promotion of cancer growth, promotion of metastasis and other side effects of chemotherapy or radiotherapy. Although cellular senescence suppresses tumorigenesis early in life, studies have shown that it may promote cancer in aged organisms (Krtolica et al; PNAS, Oct.9, 2001, Vol 98, No 21, 12072-12077). Oncogene-induced senescence is classically considered a tumor defense barrier. However, several studies have shown that under certain circumstances, senescent cells may favor tumor progression because of their secretory phenotype (Angelini et al; Cancer Res 73(1), 1 Jan.2013, 450-458).

[0080] In certain embodiments, the described compositions can decrease inflammation that drives cancer and metastasis.

[0081] In certain embodiments, the described compositions can be used in the treatment of a chemotherapy-induced or radiotherapy-induced side effect. Non-limiting examples of chemotherapeutic agents include anthracyclines, doxorubicin, etoposide, daunorubicin, taxols, paclitaxel, gemcitabine, pomalidomide, and lenalidomide. Chemotherapy-induced side effects or radiotherapy-induced side effects include, but art not limited to, weight loss, endocrine changes, hormone imbalance, changes in hormone signaling, changes is cardiotoxicity, body composition, reduced ability to be physically active, gastrointestinal toxicity, nausea, vomiting, constipation, anorexia, diarrhea, peripheral neuropathy, fatigue, malaise, low physical activity, hematological toxicity, anemia, hepatotoxicity, alopecia, pain, infection, mucositis, fluid retention, dermatological toxicity, rashes, dermatitis, hyperpigmentation, urticaria, photosensitivity, nail changes, mouth, gum or throat problems, and any toxic side effect caused by a chemotherapy or radiotherapy. Thus, in certain embodiments, the described compositions can be used in combination treatment with a chemotherapeutic agent, where the agent is administered separately, sequentially or concomitantly with the chemotherapeutic agent.Likewise, the compositions can be used in combination treatment with radiotherapy, where the agent is administered before, during or after radiotherapy. Another embodiment of the invention relates to a composition as described herein for use in reducing or alleviating one or more chemotherapy-induced or radiotherapy-induced side effects.

[0082] In certain embodiments, described herein are methods for treating or reducing the likelihood of metastasis comprising administering a composition described herein during an off-chemotherapy or off-radiotherapy time interval or after the chemotherapy or radiotherapy treatment regimen has been completed. Another embodiment relates to a composition as described herein for use in treating or reducing the likelihood of metastasis.

[0083] In certain embodiments, the described compositions can be used in the treatment of chronic or long-term chemotherapy-induced or radiotherapy-induced side effects. Certain toxic effects can appear long after treatment and can result from damage to an organ or system by the therapy. Organ dysfunction, for example, neurological, pulmonary, cardiovascular, and endocrine dysfunction, can be observed in subjects who were treated for cancers during childhood. Chronic or late toxic side effects that occur in subjects who received chemotherapy or radiation therapy include, for example, cardiomyopathy, congestive heart disease, inflammation, early menopause, osteoporosis, infertility, impaired cognitive function, peripheral neuropathy, secondary cancers, cataracts and other vision problems, hearing loss, chronic fatigue, reduced lung capacity, and lung disease.

[0084] Cardiovascular Disease

[0085] In certain embodiments, the senescence-associated disease or disorder is a cardiovascular disease. Examples include, but are not limited to, atherosclerosis, angina, arrhythmia, cardiomyopathy, congestive heart failure, coronary artery disease, carotid artery disease, endocarditis, coronary thrombosis, myocardial infarction, hypertension, aortic aneurysm, cardiac diastolic dysfunction, hypercholesterolemia, hyperlipidemia, mitral valve prolapsed, peripheral vascular disease, cardiac stress resistance, cardiac fibrosis, brain aneurysm and stroke.

[0086] In certain embodiments, the senescence-associated disease or disorder is associated with or caused by atherosclerosis (i.e. hardening of the arteries).Atherosclerosis is characterized by patchy intimal plaques (atheromas) that encroach on the lumen of medium-sized and large arteries. Administration of the described composition can reduce the lipid content of an atherosclerotic plaque in a blood vessel of the subject and / or increase the fibrous cap thickness.

[0087] Inflammatory and Autoimmune Diseases and Disorders

[0088] In certain embodiments, the senescence-associated disease or disorder is an inflammatory or autoimmune disease or disorder. Non-limiting examples of autoimmune diseases include osteoporosis, osteoarthritis, psoriasis, oral mucositis, rheumatoid arthritis, inflammatory bowel disease, eczema, kyphosis (curvature of the spinal column), herniated intervertebral disc, and the pulmonary diseases, COPD and idiopathic pulmonary fibrosis. In one embodiment, the senescence-associated disease or disorder is chronic inflammation.

[0089] Chronic inflammation is thought to be the main age-related factor associated with osteoarthritis. In combination with aging, joint overuse and obesity appear to promote osteoarthritis. By selectively killing senescent cell, the described composition can reduce or inhibit loss or erosion of proteoglycan layers in a joint, reduce inflammation in the affected joint, and promote production of collagen. Removal of senescent cells causes a reduction in the amount of inflammatory cytokines, such as IL-6, produced in a joint and inflammation is reduced.

[0090] In certain embodiments, the senescence-associated disease or disorder is rheumatoid arthritis. Rheumatoid arthritis is a chronic inflammatory disorder that typically affects the joints in hands and feet.

[0091] In certain embodiments, the senescence-associated disease or disorder is osteoporosis. Osteoporosis is a progressive bone disease that is characterized by a decrease in bone mass and density that may lead to an increased risk of fracture. Bone mineral density (BMD) is reduced, bone microarchitecture deteriorates, and the amount and variety of proteins in bone are altered.

[0092] In certain embodiments, the described compositions is for use in treating herniated intervertebral discs. Subjects with herniated discs exhibit elevated presence of cell senescence in the blood and in vessel walls (Roberts et al. (2006) Eur. Spine J.15 Suppl 3: S312-316).

[0093] Neurological Disease or Disorder

[0094] In certain embodiments, the senescence-associated disease or disorder is a neurological disease or disorder selected from Alzheimer's disease (and other dementias), Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment (MCI), macular degeneration and motor neuron dysfunction (MND), and diseases and disorders of the eyes, such as age-related macular degeneration.

[0095] Parkinson's disease (PD) is a disabling condition of the brain characterized by slowness of movement (bradykinesia), shaking, stiffness, postural instability and loss of balance. Many of these symptoms are due to the loss of certain nerves in the brain, which results in a lack of dopamine. Senescence of dopamine-producing neurons is thought to contribute to the observed cell death in PD through the production of reactive oxygen species (Cohen et al, J. Neural Transm. Suppl.19:89-103 (1983)).

[0096] Alzheimer's disease (AD) is a neurodegenerative disease that shows a slow progressive mental deterioration with failure of memory, disorientation, and confusion, leading to profound dementia. As the disease progresses, impaired judgment, confusion, behavioral changes, disorientation, and difficulty in walking and swallowing occur. Age is the single greatest predisposing risk factor for developing AD, which is the leading cause of dementia in the elderly (Hebert, et al., Arch. Neurol.60:1119-1122 (2003)). Early clinical symptoms show remarkable similarity to mild cognitive impairment.

[0097] Mild Cognitive Impairment (MCI) is a brain-function syndrome involving the onset and evolution of cognitive impairments beyond those expected based on age and education of the individual, but which are not significant enough to interfere with the individual's daily activities. MCI is an aspect of cognitive aging that is considered to be a transitional state between normal aging and the dementia into which it may convert (Pepeu, Dialogues in Clinical Neuroscience 6:369-377, 2004). MCI that primarily affects memory is known as “amnestic MCI,” which is frequently seen as prodromal stage of Alzheimer's disease. MCI that affects thinking skills other than memory is known as “non-amnestic MCI.”

[0098] MND is a group of progressive neurological disorders that destroy motor neurons, the cells that control voluntary muscle activities such as speaking, walking, breathing and swallowing. Examples of MNDs include Amyotrophic Lateral Sclerosis(ALS), also known as Lou Gehrig's Disease, progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, progressive muscular atrophy, lower motor neuron disease, and spinal muscular atrophy (SMA) (e.g., SMA1 also called Werdnig-Hoffmann Disease, SMA2, SMA3 also called Kugelberg-Welander Disease, and Kennedy's disease), post-polio syndrome, and hereditary spastic paraplegia.

[0099] Ophthalmic Diseases and Disorders

[0100] In certain embodiments, the senescence-associated disease or disorder is an ocular disease, disorder, or condition. Examples include, but are not limited to, presbyopia, macular degeneration, cataracts and glaucoma. Macular degeneration is a neurodegenerative disease that causes the loss of photoreceptor cells in the central part of retina, called the macula.

[0101] Metabolic Disease

[0102] In certain embodiments, the senescence-associated disease or disorder is a metabolic disease selected from diabetes, diabetic ulcer, metabolic syndrome, and obesity.

[0103] Senescent cells are understood to play a role in metabolic diseases, such as obesity and type 2 diabetes. Studies have shown that fat tissue from obese mice showed induction of the senescence markers SA-^-Gal, p53, and p21 (Tchkonia et al, Aging Cell 9:667-684 (2010); Minamino et al, Nat. Med.15: 1082-1087 (2009)). Induction of senescent cells in obesity has potential clinical implications because pro-inflammatory SASP components are also believed to contribute to type 2 diabetes. A similar pattern of up-regulation of senescence markers and SASP components are associated with diabetes, both in mice and in humans. Accordingly, the agents described herein have potential applications in treating or preventing type 2 diabetes, obesity and metabolic syndrome.

[0104] Pulmonary Disease or Disorder

[0105] In certain embodiments, the senescence-associated disease or disorder is a pulmonary disease. Examples include, but are not limited to, pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), asthma, cystic fibrosis, emphysema, bronchiectasis, and age-related loss of pulmonary function.

[0106] COPD is a lung disease defined by persistently poor airflow resulting from the breakdown of lung tissue (emphysema) and the dysfunction of the small airways(obstructive bronchiolitis). Pulmonary fibrosis is a chronic and progressive lung disease characterized by stiffening and scarring of the lung, which may lead to respiratory failure, lung cancer, and heart failure. The described compositions can also be used for treating a subject who is aging and has loss (or degeneration) of pulmonary function (i.e., declining or impaired pulmonary function compared with a younger subject) and / or degeneration of pulmonary tissue.

[0107] Other Age-Related Disorders

[0108] In certain embodiments, the senescence-associated disease or disorder is an age-related disorder selected from renal disease, renal failure, frailty, hearing loss, muscle fatigue and / or loss, skin conditions (e.g., wrinkles, dark spots, age-spots, etc.), skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis, and sarcopenia.

[0109] Dermatological Diseases or Disorders

[0110] In certain embodiments, the senescence-associated disease or disorder is a dermatological disease or disorder. Examples include, but are not limited to, eczema, psoriasis, hyperpigmentation, nevi, rashes, atopic dermatitis (a form of eczema and associated with inflammation), urticaria, diseases and disorders related to photosensitivity or photoaging, rhytides (wrinkles due to aging); pruritis (linked to diabetes and aging); dysesthesia (chemotherapy side effect linked to diabetes and multiple sclerosis); eczematous eruptions (often observed in aging patients and linked to side effects of certain drugs); eosinophilic dermatosis (linked to certain kinds of hemotologic cancers); reactive neutrophilic dermatosis (associated with underlying diseases such as inflammatory bowel syndrome); pemphigus; pemphigoid; immunobullous dermatosis (autoimmune blistering of skin); fibrohistocytic proliferations of skin; cutaneous lymphomas; and cutaneous lupus. Late onset lupus may be linked to decreased (i.e., reduced) function of T-cell and B-cells and cytokines (immunosenescence) associated with aging.

[0111] Lifespan and Age-Related Diseases or Conditions

[0112] In certain embodiments, the described compositions can be used to extend the lifespan of a subject by selectively killing senescent cells over non-senescent cells. In some embodiments, extending the lifespan of the subject comprises delaying onset or progression of an age-related disease or condition. In certain embodiments, the age-related disease or condition is selected from atherosclerosis, cardiovascular disease, cancer, arthritis, dementia, cataract, osteoporosis, diabetes, hypertension, age-related fat loss, lipodystrophy, and kidney disease. In some embodiments, the age-related disease or condition is geriatric anxiety disorder. In some embodiments, the age-related disease or condition is age-related inactivity. In some embodiments, the age-related disease or condition is reduction of spontaneous activity. In some embodiments, the age-related disease or condition is reduction of exploratory behavior.

[0113] In some embodiments, the onset or progression of an age-related disease or condition can be delayed by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year. In one embodiment, the described composition is for use in improving the physical health of a subject. In another preferred embodiment, the described composition is for use in reverting ageing-associated changes of metabolites in the blood.

[0114] Diseases Associated with Elevated ^-Galactosidase Activity

[0115] In another embodiment, the composition can be used in the treatment of a disease or disorder which correlates, or is associated with, elevated ^-galactosidase activity. In a preferred embodiment, the elevated ^-galactosidase activity is a result of elevated expression of ^-galactosidase or overexpression of ^-galactosidase activity relative to baseline levels which can be determined by standard methods.

[0116] Expression of ^-galactosidase can be detected in cells by histochemical or immunohistochemcal methods. For example, the senescence marker SA-beta galactosidase (SA-^-Gal) can be detected by known methods (Dimri et al, Proc. Natl. Acad. Sci. USA 92: 9363-9367 (1995).

[0117] In another embodiment, the disease is selected from the Wiedemann- Rautenstrauch syndrome of neonatal progeria, the Werner syndrome of adult progeria, Hutchinson-Gilford syndrome, Rothmund Thompson syndrome, Mulvill-Smith syndrome, Cockayne syndrome, Dyskeratosis Congenita, idiopathic pulmonary fibrosis, aplastic anemia, emphysema, type 2 diabetes, and degeneration of cartilage.

[0118] Pharmaceutical Compositions / Formulations

[0119] For use according to the described embodiments, the described ingredients or extracts may be presented as a pharmaceutical formulation with one or more pharmaceutically acceptable excipients and / or carriers therefore and optionally other therapeutic and / or prophylactic ingredients. The excipient(s) and / or carrier(s) must be acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. The pharmaceutical compositions, neutraceutical compositions, and cosmetic compositions may be for human or animal usage in human and veterinary medicine.

[0120] Examples of such suitable excipients for the various different forms of pharmaceutical compositions described herein may be found in the “Handbook of Pharmaceutical Excipients, 2ndEdition, (1994), Edited by A Wade and P J Weller. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit.1985).

[0121] Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like. Examples of suitable diluents include ethanol, glycerol and water.

[0122] The choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice. The pharmaceutical compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilizing agent(s), buffer(s), flavouring agent(s), surface active agent(s), thickener(s), preservative(s) (including anti-oxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.

[0123] Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flow lactose, beta-lactose, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol.

[0124] Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents may be also used.

[0125] Pharmaceutical formulations include those suitable for oral, topical (including dermal, buccal and sublingual), rectal or parenteral (including subcutaneous, intradermal, intramuscular and intravenous), nasal and pulmonary administration e.g., by inhalation.

[0126] The formulation may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association an active compound with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.

[0127] Pharmaceutical formulations suitable for oral administration wherein the carrier is a solid are most preferably presented as unit dose formulations such as boluses, capsules or tablets each containing a predetermined amount of the extract of Camellia nitidissima Chi. A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine an active compound in a free-flowing form such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, lubricating agent, surface-active agent or dispersing agent. Molded tablets may be made by molding an active compound with an inert liquid diluent. Tablets may be optionally coated and, if uncoated, may optionally be scored. Capsules may be prepared by filling an active compound, either alone or in admixture with one or more accessory ingredients, into the capsule shells and then sealing them in the usual manner. Cachets are analogous to capsules wherein an active compound together with any accessory ingredient(s) is sealed in a rice paper envelope. The described compositions may also be formulated as dispersible granules, which may for example be suspended in water before administration, or sprinkled on food. The granules may be packaged, e.g., in a sachet. Formulations suitable for oral administration, wherein the carrier is a liquid may be presented as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water liquid emulsion.

[0128] Formulations for oral administration include controlled release dosage forms, e.g. tablets wherein an agent is formulated in an appropriate release-controlling matrix, or is coated with a suitable release-controlling film. Such formulations may be particularly convenient for prophylactic use. Other formulations for oral administration may be in a form of a dietary supplement (i.e., food supplement).

[0129] A “dietary supplement” is a manufactured product intended to supplement one's diet by taking a pill, capsule, tablet, powder, or liquid. A supplement described herein can provide nutrients extracted from Camellia nitidissima Chi. The class of nutrient compounds includes vitamins, minerals, fiber, fatty acids, and / or amino acids. Dietary supplements described herein can also contain substances that have not been confirmed as being essential to life, but are marketed as having a beneficial biological effect, such as plant pigments or polyphenols. These supplements may be sold individually and in combination, and may be combined with nutrient ingredients.

[0130] Pharmaceutical formulations suitable for rectal administration wherein the carrier is a solid are most preferably presented as unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by admixture of an active compound with the softened or melted carrier(s) followed by chilling and shaping in molds. Pharmaceutical formulations suitable for parenteral administration include sterile solutions or suspensions of an active compound in aqueous or oleaginous vehicles.

[0131] Injectable preparations may be adapted for bolus injection or continuous infusion. Such preparations are conveniently presented in unit dose or multi-dose containers which are sealed after introduction of the formulation until required for use. Alternatively, the extract of Camellia nitidissima Chi may be in powder form which is constituted with a suitable vehicle, such as sterile, pyrogen-free water, before use.

[0132] The described compositions may also be formulated as long-acting depot preparations, which may be administered by intramuscular injection or by implantation, e.g., subcutaneously or intramuscularly. Depot preparations may include, for example, suitable polymeric or hydrophobic materials, or ion-exchange resins. Such long-acting formulations are particularly convenient for prophylactic use.

[0133] Formulations suitable for pulmonary administration via the buccal cavity are presented such that particles containing an active compound and desirably having a diameter in the range of 0.5 to 7 microns are delivered in the bronchial tree of the recipient.

[0134] As one possibility, such formulations are in the form of finely comminuted powders which may conveniently be presented either in a pierceable capsule, suitably of, for example, gelatin, for use in an inhalation device, or alternatively as a self-propelling formulation comprising an active compound, a suitable liquid or gaseous propellant and optionally other ingredients such as a surfactant and / or a solid diluent. Suitable liquid propellants include propane and the chlorofluorocarbons, and suitable gaseous propellants include carbon dioxide. Self-propelling formulations may also be employed wherein an active compound is dispensed in the form of droplets of solution or suspension.

[0135] Such self-propelling formulations are analogous to those known in the art and may be prepared by established procedures. Suitably they are presented in a container provided with either a manually-operable or automatically functioning valve having the desired spray characteristics; advantageously the valve is of a metered type delivering a fixed volume, for example, 25 to 100 microliters, upon each operation thereof. As a further possibility, an agent may be in the form of a solution or suspension for use in an atomizer or nebulizer whereby an accelerated airstream or ultrasonic agitation is employed to produce a fine droplet mist for inhalation.

[0136] Formulations suitable for nasal administration include preparations generally similar to those described above for pulmonary administration. When dispensed such formulations should desirably have a particle diameter in the range 10 to 200 microns to enable retention in the nasal cavity; this may be achieved by, as appropriate, use of a powder of a suitable particle size or choice of an appropriate valve. Other suitable formulations include coarse powders having a particle diameter in the range 20 to 500 microns, for administration by rapid inhalation through the nasal passage from a container held close up to the nose, and nasal drops comprising 0.2 to 5% w / v of an active compound in aqueous or oily solution or suspension.

[0137] Pharmaceutically acceptable carriers are well known to those skilled in the art and include, but are not limited to, 0.1 M and preferably 0.05 M phosphate buffer or0.8% saline. Additionally, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents, inert gases and the like.

[0138] Formulations suitable for topical administration may be provided for example as gels, creams or ointments. Such preparations may be applied to skin either directly spread upon the surface of the skin or carried on a suitable support such as a bandage, gauze, mesh or the like which may be applied to and over the area to be treated.

[0139] Liquid or powder formulations may also be provided which can be sprayed or sprinkled directly onto the site to be treated. Alternatively, a carrier such as a bandage, gauze, mesh or the like can be sprayed or sprinkle with the formulation and then applied to the site to be treated.

[0140] Pharmaceutical or veterinary composition as described above can be prepared by bringing the agent into association with the carrier, for example by admixture.

[0141] In general, the formulations are prepared by uniformly and intimately bringing into association the agent with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product.

[0142] In certain embodiments, the described compositions may be formulated with a neutraceutical excipient.

[0143] In certain other embodiments, the described compositions may be formulated with a cosmeceutical carrier or excipient. Cosmetically acceptable carrier or excipient may be water, a glycerin, a C1-C4 alcohol, a fatty alcohol, a fatty ether, a fatty ester, a polyol, a glycol, a vegetable oil, a mineral oil, a liposome, a laminar lipid material, a silicone oil, or combinations thereof.

[0144] Administration

[0145] The compositions described herein may be adapted for any suitable and known administration. Preferably the formulation is an orally administered formulation. The formulations may conveniently be presented in unit dosage form, i.e., in the form of discrete portions containing a unit dose, or a multiple or sub-unit of a unit dose. By way of example, the formulations may be in the form of tablets and sustained release capsules, and may be prepared by any method well known in the art of pharmacy.

[0146] Formulations for oral administration in the present invention may be presented as: discrete units such as capsules / gelules, drops, cachets, pills or tablets each containing a predetermined amount of the active agent; as a powder or granules; as a solution, emulsion or a suspension of the active agent in an aqueous liquid or a non- aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion; or as a bolus etc. Preferably, these compositions contain from 1 to 250 mg and more preferably from 10-100 mg, of active ingredient per dose.

[0147] For compositions for oral administration (e.g. tablets and capsules), the term “acceptable carrier” includes vehicles such as common excipients e.g. binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (Povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropyl-methylcellulose, sucrose and starch; fillers and carriers, for example corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride and alginic acid; and lubricants such as magnesium stearate, sodium stearate and other metallic stearates, glycerol stearate stearic acid, silicone fluid, talc waxes, oils and colloidal silica. Flavouring agents such as peppermint, oil of wintergreen, cherry flavouring and the like can also be used. It may be desirable to add a coloring agent to make the dosage form readily identifiable. Tablets may also be coated by methods well known in the art.

[0148] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active agent in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tabletsmay be optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active agent.

[0149] Other formulations suitable for oral administration include lozenges comprising the active agent in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active agent in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active agent in a suitable liquid carrier.

[0150] Other forms of administration comprise solutions or emulsions which may be injected intravenously, intraarterially, intrathecally, subcutaneously, intradermally, intraperitoneally or intramuscularly, and which are prepared from sterile or sterilizable solutions. Injectable forms typically contain between 10-1000 mg, preferably between 10- 250 mg, of active ingredient per dose.

[0151] The described compositions may also be in form of suppositories, pessaries, suspensions, emulsions, lotions, ointments, creams, gels, sprays, solutions or dusting powders.

[0152] An alternative means of transdermal administration is by use of a skin patch. For example, the active ingredient can be incorporated into a cream consisting of an aqueous emulsion of polyethylene glycols or liquid paraffin. The active ingredient can also be incorporated, at a concentration of between 1 and 10% by weight, into an ointment consisting of a white wax or white soft paraffin base together with such stabilizers and preservatives as may be required.

[0153] Combinations

[0154] In a particularly preferred embodiment, the described compositions may be administered in combination with one or more other active agents, for example, existing drugs available on the market. In such cases, the described compositions may be administered consecutively, simultaneously or sequentially with the one or more other active agents. Drugs in general are more effective when used in combination. In particular, combination therapy is desirable in order to avoid an overlap of major toxicities, mechanism of action and resistance mechanism(s). Furthermore, it is also desirable to administer most drugs at their maximum tolerated doses with minimum time intervals between such doses. The major advantages of combining chemotherapeuticdrugs are that it may promote additive or possible synergistic effects through biochemical interactions and also may decrease or delay the emergence of resistance. Beneficial combinations may be suggested by studying the activity of the test compounds with agents known or suspected of being valuable in the treatment of a particular disorder.

[0155] In certain embodiments, the composition may further include additional ingredients, extract and / or actives.

[0156] For example, in certain embodiments, the described compositions may include an ingredient or an extract of Lycium ruthenicum (also referred to as Black Goji). Lycium ruthenicum (Chinese:pinyin: hei guo gou qi), is a flowering plant also known as Russian box thorn, black fruit wolfberry, siyah goji, and kaokee. The fruits of Lycium ruthenicum contain an immunologically active pectin, which is known to improve immune function against chronic diseases.

[0157] Dosage

[0158] A person of ordinary skill in the art can easily determine an appropriate dose of one of the instant compositions to administer to a subject without undue experimentation. Typically, a physician will determine the actual dosage which will be most suitable for an individual patient and it will depend on a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual undergoing therapy. The dosages disclosed herein are exemplary of the average case. There can of course be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

[0159] The described compositions may also be administered orally to the patient, in a manner such that the concentration of drug is sufficient to achieve one or more of the therapeutic indications disclosed herein. Typically, a composition containing the extract of leaves of Camellia nitidissima Chi is administered at an oral dose of about ^20g / day. The yield of leaves is about 8%. In certain embodiments, the leaf extract of Camellia nitidissima Chi may be administered at an oral dose of between about 0.1 to about 26.67 mg / kg (60 kg per person).

[0160] No unacceptable toxicological effects are expected when described compositions are administered in accordance with the present embodiments. The described extracts of Camellia nitidissima Chi, which may have good bioavailability, may be tested in one of several biological assays to determine the concentration of a compound which is required to have a given pharmacological effect.

[0161] Methods

[0162] Certain embodiments relate to a method for treating, preventing or reversing aging or age-related disease, disorder, or condition in a subject, comprising administering to the subject a therapeutically effective amount of an alcohol or a water extract of the leaves of Camellia nitidissima Chi and a pharmaceutically acceptable excipient. The age-related disease, disorder, or condition may be skin conditions, skin wound healing, renal disease, renal failure, frailty, hearing loss, muscle fatigue, muscle loss, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia. Other diseases, disorders, or conditions mentioned throughout this application are also contemplated.

[0163] Another embodiment relates to a method for selectively killing one or more senescent cells in a subject, comprising administering to the subject a therapeutically effective amount of an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically, acceptable excipient.

[0164] Treatment regimen

[0165] The treatment regimen of the methods for treating a senescence associated disease or disorder, comprises administering the described compositions for a time sufficient and in an amount sufficient that selectively kills senescent cells. In certain embodiments, the described compositions may be administered within a treatment cycle, which treatment cycle comprises a treatment course followed by a non-treatment interval. A treatment course of administration refers herein to a finite time frame over which one or more doses of the described compositions on one or more days are administered. The finite time frame may be also called herein a treatment window.

[0166] In one embodiment, a method for treating a senescence-associated disease or disorder, is described, and which method comprises administering to a subject in need thereof the described composition that selectively kills senescent cells and is administeredwithin a treatment cycle. In a particular embodiment, the methods comprise administering the described composition in at least two treatment cycles. In a specific embodiment, the non-treatment interval may be at least about 2 weeks or between at least about 0.5-12 months, such as at least about one month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 12 months (i.e., 1 year). In other certain particular embodiments, the non-treatment interval is between 1-2 years or between 1-3 years, or longer. In certain embodiments, each treatment course is no longer than about 1 month, no longer than about 2 months, or no longer than about 3 months; or is no longer than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 26, 27, 28, 29, 30, or 31 days.

[0167] In certain embodiments, the treatment window (i.e., treatment course) is only one day. In other certain embodiments, a single treatment course occurs over no longer than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 26, 27, 28, 29, 30, or 31 days. During such treatment windows, the described composition may be administered at least on two days (i.e., two days or more) with a variable number of days on which the composition is not administered between the at least two days of administration. Stated another way, within a treatment course when the composition is administered on two or more days, the treatment course may have one or more intervals of one or more days when the composition, is not administered. By way of non-limiting example, when the composition is administered on 2 or more days during a treatment course not to exceed 21 days, the composition may be administered on any total number of days between from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 26, 27, 28, 29, 30, or 31 days.

[0168] In certain embodiments, the treatment course comprises a length of time during which the composition may be administered daily. In one specific embodiment, the composition is administered daily for 2 days. In another specific embodiment, the composition is administered daily for 3 days. In yet another particular embodiment, the composition is administered daily for 4 days. In one specific embodiment, the composition is administered daily for 5 days. In yet another particular embodiment,the composition is administered daily for 6 days. In another specific embodiment, the composition is administered daily for 7 days. In yet another particular embodiment, the composition is administered daily for 8 days. In still another specific embodiment, the composition is administered daily for 9 days. In yet another particular embodiment, the composition is administered daily for 10 days. In yet another particular embodiment, the composition is administered daily for 11 days. In yet another particular embodiment, the composition is administered daily for 12 days. In yet another particular embodiment, the composition is administered daily for 13 days. In yet another particular embodiment, the composition is administered daily for 14 days. The treatment window (i.e., course) for each of the above examples is no longer than 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, respectively.

[0169] The invention is further described by way of the following non-limiting examples.

[0170] EXAMPLES

[0171] Example 1:

[0172] ^^^ ^^^^^^^^^^^^^^^^^^^^

[0173] (1) Extract 1: Dried leaves of Camellia nitidissima Chi (50 g) were refluxed in 500 mL of water solution at 95 ºC in a water bath for 1.5 h and then in 350 mL of fresh solution for a further 1 h. The water extracts were combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure.

[0174] (2) Extract 2: For alcoholic extract, dried leaves of Camellia nitidissima Chi (50 g) were refluxed in 500 mL of 70% (v / v) ethanol solution at 95 ºC in a water bath for 1.5 h and then in 350 mL of fresh solution for a further 1 h. The ethanol extracts were combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure.

[0175] (3) Extract 3: In order to make alcoholic extract of flower, dried flowers of Camellia nitidissima Chi (50 g) were refluxed in 600 mL of 70% (v / v) ethanol solution at 95 ºC in a water bath for 1.5 h and then in 400 mL of fresh solution for a further 1 h. The ethanol extracts were combined, filtered, and then concentrated using rotary vaporization at 60°C under reduced pressure.

[0176] These three extract-powders were obtained by drying the concentrated solution using a freeze dryer and stored in a refrigerator at -20°C until use.

[0177] II. Cell viability of leaves water extract on Hs27 human fibroblasts.

[0178] Cells were treated with 6 concentrations (15.63, 31.25, 62.50, 125.00, 250.00 and 500,00 µg / ml) for 72 h. Cell proliferation was determined by CCK8 assay. Figure 1 is a JHCY-W (leaves water extract) graph of cell viability of leaves water extract on Hs27 human fibroblasts. Values in Figure 1 are represented as the means ± SD.15.63 µg / ml was used as max concentration for the next testing since the cell viability was higher than 80%.

[0179] III. Cell viability of leaves alcoholic extract on Hs27 human fibroblasts.

[0180] Cells were treated with 6 concentrations (15.63, 31.25, 62.50, 125.00, 250.00 and 500,00 µg / ml) for 72 h. Cell proliferation was determined by CCK8 assay. Values are represented as the means ± SD. Figure 2 is a JHCY-E (leaves alcoholic extract) graph of cell viability of leaves alcoholic extract on Hs27 human fibroblasts. Values in Figure 2 are represented as the means ± SD.15.63 µg / ml was used as max concentration for the next testing since the cell viability is higher than 80%.

[0181] IV. Anti-aging function of either water or alcoholic extract from leaves.

[0182] Figure 3 shows: A. D-galactose was used to generate a model of accelerated aging on Hs27. Cells were treated either water or alcoholic extracts from leaves. Aging cell were stained by SA-^-Gal positivity after 72 h treatment (blue color). B. Aging cell ratio after treatment of different concentrations of water extract from leaves. Values are represented as the means ± SE. ** indicates p < 0.001 compared with Model group. C. Aging cell ratio after treatment of different concentrations of alcoholic extract from leaves. Values are represented as the means ± SE. ** indicates p < 0.001 compared with Model group.

[0183] V. Senolytic potential of alcoholic extract from flowers.

[0184] To validate the role of the compositions described herein as senolytics, the extracts of Camellia nitidissima Chi were tested for their ability to selectively eliminate aging cells at different concentrations.

[0185] Figure 4: (A) Mitomycin C was used to generate a model of accelerated aging on WI-38. SA-^-Gal staining kit was used to check aging. Remove growth mediafrom the cells. Rinse the plate one time with PBS. Add Fixative Solution and fix cells for 10-15 min at room temperature. The plate was rinsed two times with PBS. ^- Galactosidase Staining Solution was added to cells and the cells were incubated at 37°C at least overnight in a dry incubator (no CO2). The ^-Galactosidase staining solution was removed and the cells were overlaid with PBS. Aging cell were stained by SA-^-Gal positivity (black dots). (B) Both aging and normal cells were digested and seeded in 384-well plate. After cell adherence, both aging and normal cells were treated with different dosages of alcoholic extract from flowers for 7-day treatment. Cell viabilities were detected after treatments ratio after treatment by CellTiter-Glo assay.

[0186] VI. Conclusions

[0187] The water and alcoholic extracts of leaves of Camellia nitidissima Chi have the potential to reverse aging (Figure 3). After treatments, the ratio of aging cells was decreased. Besides, flower alcoholic extract has senolytic activity (Figure 4). Senolytic agents can be used to selectively induce programmed death of senescent cells. Aging cells treated with alcoholic extract of flowers are more sensitive to die compared to those normal cells treated under the same concentration.

Claims

CLAIMS 1. A composition for use in treating, preventing or reversing aging or age-related disease, disorder or condition, comprising an alcohol or a water extract of leaves of Camellia nitidissima Chi and a pharmaceutically acceptable excipient.

2. The composition of claim 1, wherein the age-related disease, disorder, or condition is skin conditions, skin wound healing, renal disease, renal failure, frailty, hearing loss, muscle fatigue, muscle loss, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia.

3. The composition of any of claims 1-2, wherein the composition is formulated for oral administration.

4. The composition of any of claims 1-3, wherein the composition is formulated for a topical administration.

5. The composition of claim 4, wherein the composition is in a form of an aerosol, a cream, a emulsion, a solid, a liquid, a dispersion, a foam, a gel, a lotion, a mousse, an ointment, a powder, a patch, a pomade, a solution, a pump a spray, a stick, am oil, a towelette, or combinations thereof.

6. The composition of any of claims 1-5, wherein the alcohol or water extract of Camellia nitidissima Chi is present in an amount of between about 0.1 to about 26.67 mg / kg (60 kg per person).

7. The composition of any of claims 1-6, further comprising an extract of Lycium ruthenicum.

8. A composition for use in selectively killing one or more senescent cells, comprising an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically acceptable excipient.

9. A composition for use in treating or preventing a senescence-associated disease or disorder, comprising an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically acceptable excipient.

10. The composition of claim 9, wherein the senescence-associated disease or disorder is a cardiovascular disease or disorder, a metabolic disease, an inflammatory disease or disorder, a pulmonary disease or disorder, a neurological disease or disorder, a renal disorder or disease, or a dermatological disorder or disease.

11. The composition of any of claims 9-10, wherein the senescence-associated disease or disorder is selected from: (i) an inflammatory or autoimmune disease or disorder selected from osteoarthritis, osteoporosis, oral mucositis, inflammatory bowel disease, kyphosis and herniated intervertebral disc; (ii) a neurological disease or disorder selected from Alzheimer's disease, Parkinson's disease, Huntington's disease, dementia, mild cognitive impairment, macular degeneration and motor neuron dysfunction; (iii) a metabolic disease selected from diabetes, diabetic ulcer, metabolic syndrome and obesity; (iv) a pulmonary disease selected from pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema, bronchiectasis and age-related loss of pulmonary function; (v) an eye disease or disorder selected from macular degeneration, glaucoma, cataracts, presbyopia and vision loss;(vi) an age-related disorder selected from renal disease, renal failure, frailty, hearing loss, muscle fatigue, skin conditions, skin wound healing, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia; (vii) a dermatological disease or disorder selected from eczema, psoriasis, hyperpigmentation, nevi, rashes, atopic dermatitis, urticaria, diseases and disorders related to photosensitivity or photoaging, rhytides, pruritis, dysesthesia, eczematous eruptions, eosinophilic dermatosis, reactive neutrophilic dermatosis, pemphigus, pemphigoid, immunobullous dermatosis, fibrohistocytic proliferations of skin, cutaneous lymphomas and cutaneous lupus; and (viii) a cardiovascular disease selected from atherosclerosis, angina, arrhythmia, cardiomyopathy, congestive heart failure, coronary artery disease, carotid artery disease, endocarditis, coronary thrombosis, myocardial infarction, hypertension, aortic aneurysm, cardiac diastolic dysfunction, hypercholesterolemia, hyperlipidemia, mitral valve prolapse, peripheral vascular disease, cardiac stress resistance, cardiac fibrosis, brain aneurysm and stroke.

12. A nutritional supplement compositing an extract of Camellia nitidissima Chi and a neutraceutically acceptable excipient.

13. The nutritional supplement of claim 12, wherein the extract is an alcohol extract or a water extract of the leaves of Camellia nitidissima Chi.

14. The nutritional supplement of claim 12, wherein the extract is an alcohol extract of the flowers of Camellia nitidissima Chi.

15. The nutritional supplement of any of claims 11-14, further comprising an extract of Lycium ruthenicum.

16. A method for treating, preventing or reversing aging or age-related disease, disorder, or condition in a subject, comprising administering to the subject a compositioncomprising a therapeutically effective amount of an alcohol or a water extract of the leaves of Camellia nitidissima Chi and a pharmaceutically acceptable excipient.

17. The method of claim 16, wherein the composition further comprises an extract of Lycium ruthenicum.

18. The method of any of claims 16-17, wherein the age-related disease, disorder, or condition is skin conditions, skin wound healing, renal disease, renal failure, frailty, hearing loss, muscle fatigue, muscle loss, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis and sarcopenia.

19. The method of any of claims 16-18, wherein the composition is formulated for oral administration.

20. The method of any of claims 16-19, wherein the composition is formulated for a topical administration.

21. A method for selectively killing one or more senescent cells in a subject, comprising administering to the subject a therapeutically effective amount of an alcohol extract of the flowers of Camellia nitidissima Chi and a pharmaceutically, acceptable excipient.

22. The method of claim 21, wherein the senescence-associated disease or disorder is a cardiovascular disease or disorder, a metabolic disease, an inflammatory disease or disorder, a pulmonary disease or disorder, a neurological disease or disorder, a renal disorder or disease, or a dermatological disorder or disease.

23. A composition for use in treating, preventing or reversing aging or age-related disease, disorder or condition, comprising: an alcohol or a water extract of leaves of Camellia nitidissima Chi; an extract of Lycium ruthenicum; and and a pharmaceutically acceptable excipient.

24. The composition of claim 23 for use as a dietary supplement.

25. The composition of claim 23 for use as a cosmetic product.

26. The composition of claim 23 for use in anti-aging. ^ ^^

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