Compositions and methods for treating ovarian aging, ovarian diseases and conditions, and symptoms thereof

Abstract

Methods for enhancing or improving fertility are disclosed. Methods for extending reproductive life span are disclosed. Methods for increasing oocyte mitochondrial mass or preventing loss of oocyte mitochondrial mass are disclosed. Methods for improving embryo development or improving fertility of embryos are disclosed. Methods for treating or preventing ovarian diseases or conditions are disclosed. Methods for reducing, preventing or delaying perimenopause, menopause or symptoms thereof are disclosed. The method comprises administering an emblica extract or a compound component of an emblica extract or a compound component having at least a 95% similarity score with a compound component of an emblica extract.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 241,429, filed September 7, 2021, entitled "Compositions and Methods for Treating Ovarian Diseases and Conditions, and Symptoms Thereof," which is incorporated by reference in its entirety for all purposes.

[0002] Technology Areas Aspects and embodiments disclosed herein relate to methods of treating diseases and conditions associated with mitochondrial dysfunction. More specifically, aspects and embodiments disclosed herein relate to methods of treating diseases and conditions associated with mitochondrial DNA depletion. [Background technology]

[0003] background Mitochondrial dysfunction, for example, mitochondrial DNA (mtDNA) depletion, is involved in many diseases and conditions, for example, mtDNA depletion syndrome, mitochondrial disease, viral infection, symptoms induced by viral infection, aging, age-related chronic diseases, reduced energy level and vitality, and other human pathologies.The basic questions about the biology of mitochondria and the biology of mtDNA and their role in such diseases and conditions remain largely unsolved.Animal models that can induce mitochondrial dysfunction and / or modulate the copy number and / or concentration of mtDNA have been developed, providing a system for investigating mitochondrial pathology and its role in disease processes.There is a need for treatment methods for diseases and conditions related to mitochondrial dysfunction, for example, mtDNA depletion. Summary of the Invention [Means for solving the problem]

[0004] overview According to one embodiment, a method for enhancing or improving fertility in a subject is provided.The method can include administering to the subject an effective amount of a composition comprising one or more of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form, or one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form.

[0005] According to one embodiment, a method for extending reproductive life span in a subject is provided.The method can include administering to the subject an effective amount of a composition comprising one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form.

[0006] According to one embodiment, a method for increasing oocyte mitochondrial mass or preventing oocyte mitochondrial mass decrease in a subject is provided.The method can include administering to the subject an effective amount of a composition comprising one or more of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of one or more of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form, or one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract, and chebula extract, or its pharmaceutically acceptable form.

[0007] According to one embodiment, there is provided a method for improving embryo development or improving embryo fertility.The method can comprise administering to the subject source of embryo or embryo a composition comprising an effective amount of one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form.

[0008] According to one embodiment, a method for treating or preventing ovarian disease or condition in a subject is provided.The method may comprise administering to the subject an effective amount of a composition comprising one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form.

[0009] According to one embodiment, a method for improving embryo development or improving fertilization rate of an embryo is provided. The method may include transferring ooplasm from a donor oocyte to a recipient oocyte to be fertilized to form an embryo. The donor oocyte may have a higher mitochondrial DNA (mtDNA) content than the recipient oocyte. At least one of the donor subject, the recipient subject, and the embryo may be administered an effective amount of a composition comprising one or more of emblica extract, fucus extract, chebula extract, or a pharmaceutically acceptable form thereof, or one or more compound components of one or more of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compounds having at least 95% similarity score with one or more compound components of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof.

[0010] According to one embodiment, a method for reducing, preventing or delaying perimenopause, menopause or its symptoms is provided.The method can include administering to a subject an effective amount of a composition comprising one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compound components of one or more of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form, or one or more compounds having at least 95% similarity score with one or more compound components of emblica extract, fucus extract and chebula extract, or its pharmaceutically acceptable form.

[0011] The method may include administering to the subject an effective amount of an emblica extract, one or more compound components of the emblica extract, or one or more compound components having a similarity score of at least 95% with the compound components of the emblica extract, or a pharma- ceutically acceptable form thereof.

[0012] The method may include administering to a subject an effective amount of a fucus extract, one or more compound components of the fucus extract, or one or more compound components having a similarity score of at least 95% with the compound components of the fucus extract, or a pharma- ceutically acceptable form thereof.

[0013] The method may include administering to the subject an effective amount of a chebula extract, one or more compound components of the chebula extract, or one or more compound components having a similarity score of at least 95% with the compound components of the chebula extract, or a pharma- ceutically acceptable form thereof.

[0014] In some embodiments, the emblica extract is derived from Emblica officinalis.

[0015] In some embodiments, the compound component of the extract of E. emblica is benzoic acid or a pharma- ceutically acceptable form thereof, substituted with 1-5 hydroxy groups and, optionally, 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups, or -CH=CH-(CH2) a -C(O)OH, where a is 0 to 5, and benzene substituted with 1 to 5 hydroxy groups, or a pharma- ceutically acceptable form thereof, or a combination of the above.

[0016] In some embodiments, the compound contained in the emblica extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, emblicanin A, emblicanin B, punigluconin, pedunculagin, punicaforin, phyllanemblin, kaempferol, ellagic acid, chebulinic acid, chebulaginic acid, punicalagin, a metabolite of any of the foregoing, a compound having at least a 95% similarity score to any of the foregoing, or a pharma- ceutically acceptable form of any of the foregoing.

[0017] In some embodiments, the fucus extract is derived from Fucus vesiculosus, Fucus serratus, Fucus spiralis, or Fucus guiryi.

[0018] In some embodiments, the compound component of the fucus extract is benzoic acid or a pharma- ceutically acceptable form thereof, substituted with 1-5 hydroxy groups and, optionally, 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups, or -CH=CH-(CH2) a -C(O)OH, where a is 0 to 5, and benzene substituted with 1 to 5 hydroxy groups, or a pharma- ceutically acceptable form thereof, or a combination of the above.

[0019] In some embodiments, the compound component of the fucus extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, fucoidan, punigluconin, pedunculagin, punicaforin, filanembrin, kaempferol, ellagic acid, chebulinic acid, chebulaginic acid, punicalagin, a metabolite of any of the foregoing, a compound having at least a 95% similarity score to any of the foregoing, or a pharma- ceutically acceptable form of any of the foregoing.

[0020] In some embodiments, the chebula extract is derived from Terminalia chebula, Terminalia arborea, or Lumnitzera racemose.

[0021] In some embodiments, the compound component of the chebula extract is benzoic acid or a pharma- ceutically acceptable form thereof substituted with 1-5 hydroxy groups and, optionally, 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups, or -CH=CH-(CH2) a -C(O)OH, where a is 0 to 5, and benzene substituted with 1 to 5 hydroxy groups, or a pharma- ceutically acceptable form thereof, or a combination of the above.

[0022] In some embodiments, the compound components of the chebula extract are gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, fucoidan, punigluconin, and pedunculagin, punicaforin, filanembrin, kaempferol, ellagic acid, chebulinic acid, chebulaginic acid, punicalagin, a metabolite of any of the foregoing, a compound having at least a 95% similarity score to any of the foregoing, or a pharma- ceutically acceptable form of any of the foregoing.

[0023] In some embodiments, the composition comprises two or more of: an effective amount of an emblica extract or a pharma- ceutically acceptable form thereof, or a compound component of an emblica extract or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% with a compound component of an emblica extract or a pharma- ceutically acceptable form thereof; an effective amount of a fucus extract or a pharma- ceutically acceptable form thereof, or a compound component of a fucus extract or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% with a compound component of a fucus extract or a pharma- ceutically acceptable form thereof; and an effective amount of a chebula extract or a pharma- ceutically acceptable form thereof, or a compound component of a chebula extract or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% with a compound component of a chebula extract or a pharma- ceutically acceptable form thereof.

[0024] In some embodiments, the composition is enriched with one or more compound components of emblica extract, or a pharma- ceutically acceptable form thereof, or a compound component having a similarity score of at least 95% with a compound component of emblica extract, or a pharma-ceutically acceptable form thereof.

[0025] In some embodiments, the composition is enriched with one or more compound components of a fucus extract, or a pharma- ceutically acceptable form thereof, or a compound component having a similarity score of at least 95% with a compound component of a fucus extract, or a pharma- ceutically acceptable form thereof.

[0026] In some embodiments, the composition is enriched with one or more compound components of a chebula extract, or a pharma- ceutically acceptable form thereof, or a compound component having a similarity score of at least 95% with a compound component of a chebula extract, or a pharma-ceutically acceptable form thereof.

[0027] In some embodiments, one or more compound components of the emblica extract, or one or more compound components having a similarity score of at least 95% with a compound component of the emblica extract, are purified, e.g., at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0028] In some embodiments, one or more compound components of a fucus extract, or one or more compound components having a similarity score of at least 95% with a compound component of a fucus extract, are purified, e.g., at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0029] In some embodiments, one or more compound components of the chebula extract, or one or more compound components having a similarity score of at least 95% with a compound component of the chebula extract, are purified, e.g., at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0030] In some embodiments, the effective amount is a therapeutically effective amount.

[0031] In some embodiments, the administration induces mitochondrial biogenesis and / or improves mitochondrial function.

[0032] In some embodiments, the effective amount or therapeutically effective amount is sufficient to induce mitochondrial biogenesis.

[0033] In some embodiments, the administration increases expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COX II.

[0034] In some embodiments, administration modulates the menstrual cycle, e.g., normalizes the menstrual cycle, increases ovulation events, and / or increases anti-Mullerian hormone (AMH) levels in the subject or recipient subject.

[0035] In some embodiments, the administration reduces the probability of aneuploidy and / or Leigh syndrome in an embryo.

[0036] In some embodiments, administration reduces or decreases the severity or frequency of at least one symptom of an ovarian disease or condition, such as, for example, dry skin or vagina, pelvic pain or cramps, inflammation, prolonged or irregular menstrual cycles, and reduced ovulation events.

[0037] In some embodiments, the composition is administered topically.

[0038] In some embodiments, the compositions are administered parenterally, for example, intravenously, intraperitoneally, or intramuscularly.

[0039] In some embodiments, the composition is administered enterally.

[0040] In some embodiments, the composition is formulated as a topical liquid, oil, cream, emulsion, or gel.

[0041] In some embodiments, the composition is formulated as a shampoo, conditioner, spray, cream, gel, balm, body wash, soap, lotion, or makeup.

[0042] In some embodiments, the compositions are formulated as parenteral liquid solutions.

[0043] In some embodiments, the composition is formulated as an enteral capsule or tablet, or as a dietary supplement or food, such as a food, food supplement, medical food, food additive, functional food, or beverage.

[0044] In some embodiments, the composition is administered locally.

[0045] In some embodiments, the composition is administered systemically.

[0046] In some embodiments, the compositions are formulated for immediate release.

[0047] In some embodiments, the composition is formulated for sustained release, eg, controlled release or sustained release.

[0048] The compositions may be administered in combination with standard of care treatments to improve fertility.

[0049] The compositions may be administered in combination with standard of care treatments for treating the ovarian disease or condition, or a symptom thereof.

[0050] The compositions may be administered in combination with one or more of clomiphene, tamoxifen, letrozole, metformin, gonadotropins, gonadotropin releasing hormones, dopamine agonists, and other hormonal treatments.

[0051] The compositions may be administered in conjunction with a surgical procedure, for example, fallopian tube surgery, laparoscopic surgery to remove or destroy cysts or submucosal fibroids, or laparoscopic ovarian drilling.

[0052] The compositions may be administered in combination with UV blockers, moisturizers, sunscreens, wrinkle creams, retinoids, alpha-hydroxy acids, beta-hydroxy acids, squalene, antioxidants, tretinoin, glycosaminoglycans (GAGs), lactic acid, malic acid, citric acid, tartaric acid, hydroquinone, kojic acid, L-ascorbic acid, licorice extract, N-acetylglucosamine, niacinamide, soy, dermal fillers or injectables such as hyaluronic acid or calcium hydroxylapatite, botulinum toxin, laser resurfacing procedures, ultrasound therapy, chemical peels such as glycolic acid peels, trichloroacetic acid or salicylic acid, or skin peeling procedures.

[0053] The composition is administered in combination with or encapsulated in a biomaterial selected from extracellular vesicles, collagen, hyaluronic acid, synthetic biomaterials (e.g., polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), polyethylene glycol (PEG)), fibrin, alginate, and composite biomaterials.

[0054] In some embodiments, the ovarian disease or condition is selected from perimenopause, menopause, endometriosis, ovarian cysts, premenopausal or postmenopausal ovarian cancer, e.g., ovarian epithelial cancer, ovarian tumors, e.g., ovarian germ cell tumors, ovarian low malignant potential tumors, and ovarian stromal tumors, polycystic ovary syndrome (PCOS), primary ovarian insufficiency (POI), and ovarian torsion.

[0055] The compositions may be administered in combination with one or more of hormone therapy, e.g., estrogen, progesterone, testosterone, or synthetic forms thereof, selective serotonin reuptake inhibitors (SSRIs), gabapentin, clonidine, hormonal contraception, gonadotropin releasing hormone (GnRH) agonists or antagonists, heat therapy, pain relief, nonsteroidal anti-inflammatory drugs (NSAIDs), e.g., ibuprofen or other pain relievers, spironolactone, eflornithine, electrolysis, chemotherapy, radiation therapy, and surgery, e.g., hysterectomy, bilateral salpingo-oophorectomy, and weight loss surgery, as well as lifestyle changes, e.g., weight loss, improved nutrition, and increased physical activity.

[0056] In some embodiments, the embryo may be produced by in vitro fertilization.

[0057] The method may further include measuring the mitochondrial content of the embryos and selecting embryos that respond with a measured value of mitochondrial content that is within a predetermined range.

[0058] The donor oocyte may be autologous.

[0059] The donor oocyte may be of the same species.

[0060] The donor oocyte may be of a different species.

[0061] In some embodiments, the ooplasmic transfer may be complete.

[0062] In some embodiments, ooplasmic transfer may be partial, for example by electrofusion or direct ooplasmic injection.

[0063] In some embodiments, transfer may be performed by modified intracytoplasmic sperm injection (ICSI).

[0064] In some embodiments, the transfer may be performed by autonomous germline mitochondrial energy transfer (AUGMENT) to an oocyte.

[0065] In some embodiments, the method may further include using CRISPR / Cas 9 gene editing technology on the mtDNA of the donor oocyte or the recipient oocyte.

[0066] The method may further include using CRISPR / Cas 9 gene editing technology on the mitochondrial DNA (mtDNA) of the subject's oocyte to be fertilized to form an embryo.

[0067] The method may further include introducing stem cell-generated mitochondrial DNA (mtDNA) into an oocyte of the subject to be fertilized to form an embryo.

[0068] The method may further comprise introducing an antioxidant into the culture medium of the embryo during development.

[0069] In some embodiments, transfer may be performed by nuclear genomic transfer, for example, oocyte spindle transfer, germinal vesicle (GV) transfer, pronuclear transfer (PNT), or polar body nuclear transfer (PBNT).

[0070] In some embodiments, the subject or recipient subject may be characterized by age-associated reduced fertility, infertility, and / or ovarian aging.

[0071] The subject or recipient subject may be 40 years of age or older.

[0072] The subject or recipient subject may be characterized by premature reproductive aging, e.g., primary ovarian insufficiency (POI), premature prolonged or irregular menstrual cycles, premature reduction in ovulatory events, premature menopause or perimenopause, and / or ovarian inflammation associated with premature reproductive aging.

[0073] The subject or recipient subject may be under 40 years of age.

[0074] The subject or recipient subject may be pre-menopausal.

[0075] The subject or recipient subject may be peri-menopausal.

[0076] The subject or recipient subject may be post-menopausal.

[0077] The subject or recipient subject may have a reduced ovarian reserve (DOR).

[0078] The subject or recipient subject may have normal ovarian reserve (NOR).

[0079] The subject or recipient subject may suffer from one or more of obesity, diabetes, chronic inflammation, hyperglycemia, autoimmune disease, and / or poor lifestyle factors such as smoking, alcohol use, drug use, exposure to chemicals (e.g., bisphenol A, advanced glycation end products (AGEs)) and / or radiation, nutritional deficiencies such as increased intake of burnt foods, increased intake of saturated fats, and / or reduced intake of fruits and vegetables, and / or reduced exercise.

[0080] The subject or recipient subject may be characterized by reduced anti-Mullerian hormone (AMH) levels, e.g., less than about 80 ng / mL, increased follicle stimulating hormone (FHL) levels, e.g., greater than about 10 IU / L, increased estradiol levels, and / or reduced antral follicle count (AFC), e.g., less than about 5-7 total follicles.

[0081] The subject or recipient subject may be undergoing or attempting in vitro fertilization (IVF).

[0082] The subject or recipient subject may be a poor responder to IVF.

[0083] The subject or recipient subject may be a normal responder to IVF.

[0084] The subject or recipient subject may suffer from reduced production of reproductive hormones, e.g., estrogen and / or progesterone, increased production of follicle stimulating hormone (FSH), prolonged or irregular menstrual cycles, uterine or vaginal atrophy, e.g., loss of endometrial glands, scattered gonadal rupture of the uterus (apoptotic bodies), acute inflammation of the uterus, and / or reduced thickness of the vaginal epithelium, reduced ovulatory events, ovarian inflammation, and / or infertility.

[0085] The subject or recipient subject may be characterized by an abnormality in the mitofusin (Mfn1) gene, an abnormality in the dynamic associated protein 1 (Drp1) gene, an abnormality in the caseinolytic peptidase P (Clpp) gene, an abnormality in the growth differentiation factor 9 (GDF9) gene, an abnormality in the fragile X mental retardation 1 (FMR1) gene, an abnormality in the transcription factor A (TFAM) gene, an abnormality in the mitochondrial DNA polymerase gamma (PolgA) gene, an abnormality in the mitochondrial inner membrane peptidase (IMP) gene, and / or an abnormality in the mitochondrial ABC transporter (MDR-1) gene.

[0086] The subject or recipient subject may be characterized by decreased gene expression of PPARy, PGC-1a, PGC-1b, ERR, NRF-1, NRF-2, SIRT1, SIRT3, SIRT4, and / or SIRT5.

[0087] The subject or recipient subject may be characterized by a decrease in AMP-activated protein kinase (AMPK) and / or PTEN-induced kinase 1 (PINK1) protein expression, and / or an increase in mammalian target of rapamycin (mTOR) protein expression.

[0088] The subject or recipient subject may have a somatic mitochondrial DNA mutation, for example, a T414G transverse mutation, in the oocyte that is fertilized to form the embryo or recipient oocyte.

[0089] The subject or recipient subject may have an inherited mitochondrial DNA (mtDNA) mutation, for example, a T414G transverse mutation, in the oocyte that is fertilized to form the embryo or recipient oocyte.

[0090] The subject or recipient subject may be characterized by reduced levels of mitochondrial DNA (mtDNA) in the oocytes that are fertilized to form the embryo or recipient oocyte, and / or reduced oocyte quality, e.g., chromosome misalignment and / or spindle abnormalities.

[0091] The subject or recipient subject may be characterized by a decrease in gene expression of superoxide dismutase (SOD) and / or an increase in the level of the Aldh3A2 enzyme.

[0092] The subject or recipient subject may be characterized by reduced mRNA expression of Prdx3, Prdx4, and / or Txn2 enzymes.

[0093] The subject or recipient subject may suffer from Turner syndrome, galactosemia, and / or fragile X syndrome.

[0094] The subject or recipient subject may be characterized by progressive external ophthalmoplegia (POE), spinocerebellar ataxia with epilepsy (SCAE), and / or mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS).

[0095] The subject or recipient subject may be characterized by a neurological condition.

[0096] The subject or recipient subject may be characterized by dysfunctional follicle formation, for example, follicle depletion or follicular dysfunction.

[0097] The subject or recipient subject may have a reduced number of tertiary follicles, eg, early antral, antral, and / or preovulatory follicles, and / or a reduced number of corpus luteum (CL) follicles.

[0098] The subject or recipient subject may be characterized by downregulation of estrogen receptor (ER) alpha and beta in the ovaries, reduced ESR1 or ESR2 gene expression, increased miR-206-3p RNA expression, reduced ovarian production of 17β-estradiol (E2), and / or an increase in lipid-laden ovarian stromal cells.

[0099] In some embodiments, the composition comprises a nanoparticle-based delivery vehicle.

[0100] In some embodiments, the composition comprises a skin permeation enhancer or is administered in combination with a skin permeation enhancer, for example a chemical or physical skin permeation enhancer.

[0101] In some embodiments, the composition comprises a mitochondrial targeting agent, or a delivery vehicle functionalized with a mitochondrial targeting agent.

[0102] In some embodiments, the compound components of the emblica extract are derived from, purified from, or isolated from an emblica extract.

[0103] In some embodiments, the compound components of the emblica extract are derived from, purified from, or isolated from a source other than the emblica extract.

[0104] In some embodiments, the compound components of the Emblica extract are synthetic.

[0105] In some embodiments, the compound components of a fucus extract are derived from, purified from, or isolated from a fucus extract.

[0106] In some embodiments, compound components of a fucus extract are derived from, purified from, or isolated from a source other than a fucus extract.

[0107] In some embodiments, the compound components of the fucus extract are synthetic.

[0108] In some embodiments, the compound components of the chebula extract are derived from, purified from, or isolated from a chebula extract.

[0109] In some embodiments, the compound components of the chebula extract are derived from, purified from, or isolated from a source other than the chebula extract.

[0110] In some embodiments, the compound components of the chebula extract are synthetic.

[0111] According to another embodiment, a kit is provided that comprises a preparation comprising an effective amount of donor mtDNA in a therapeutically acceptable carrier.The kit may comprise an effective amount of one or more of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compound components of one or more of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or a composition that comprises one or more compounds that have at least 95% similarity score with one or more compound components of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof.

[0112] The present disclosure contemplates all combinations of any one or more of the above aspects and / or embodiments, as well as combinations with any one or more of the embodiments set forth in the detailed description and any examples.

[0113] The accompanying drawings are not intended to be drawn to scale. In the drawings, each identical or nearly identical component illustrated in various figures is represented by a like numeral. For purposes of clarity, every component may not be shown in every figure. [Brief description of the drawings]

[0114] [Figure 1-1] Figure 1A shows quantification of immunofluorescence analysis of OXPHOS complex IV (COXII) in relative fluorescence units (RFU) in paraffin dorsal skin sections of control and mtDNA-depleted mice after 16 weeks of dox induction and topical emblica extract or control treatment. Data are mean ± SD (n = 3); *P < 0.05. Figure 1B shows RT-PCR analysis of dorsal skin RNA for genes encoding mtDNA from control and mtDNA-depleted mice after 16 weeks of dox induction and topical emblica extract or control treatment. [Figure 1-2] Figure 1C shows quantification of mtDNA content in dorsal skin samples from control mice (n=3) and mtDNA-depleted mice (n=3) after 16 weeks of dox induction and application of topical emblica extract or control treatment (mean ± sem; *P<0.05, Student's t-test). Figure 1D shows quantitative analysis of inflammatory cells in skin sections from control mice and mtDNA-depleted mice after 16 weeks of dox induction and application of topical emblica extract or control treatment (mean ± SD; *P≦0.05; **P≦0.01; ***P≦0.001, Student's t-test). [Diagram 2]Figure 2A shows representative photographs of (i) control mice after dox induction, (ii) mtDNA-depleted mice after dox induction, (iii) control mice after 51 days of dox induction and treatment with fucus extract, and (iv) mtDNA-depleted mice after 51 days of dox induction and treatment with fucus extract (preventive intervention) (n=3 for each group). Figure 2B shows quantification of mtDNA content (mean±sem; *P<0.05, Student's t-test) in dorsal skin samples from control mice (n=3) and mtDNA-depleted mice (n=3) after 16 weeks of dox induction and application of topical fucus extract or control treatment. [Diagram 3] FIG. 3 shows a schematic diagram of preventive and therapeutic in vivo experiments of the C. emblica extract. [Figure 4-1] FIG. 4A is a graph showing induced expression of mitochondrial complex IV subunit 2 (COXII) by various compositions, according to one embodiment. [Figure 4-2] FIG. 4B is a graph showing induced expression of mitochondrial transcription factor A (TFAM) by various compositions, according to one embodiment. [Figure 4-3] FIG. 4C is a graph showing induced expression of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1a) by various compositions, according to one embodiment. [Diagram 5] FIG. 5 is an image of an electrophoresis gel showing protein levels of mitochondrial biogenesis markers COXII, TFAM, and PGC-1a induced by various compositions, according to one embodiment. [Figure 6] 6A-6F are graphs showing COXII expression induced by administration of C. emblica extract in vitro from 6 hours to 96 hours after administration. [Figure 7] 7A-7F are graphs showing TFAM expression induced by administration of T. emblica extract in vitro from 6 hours to 96 hours after administration. [Figure 8]8A-8F are graphs showing PCG-1a expression induced by administration of C. emblica extract in vitro from 6 hours to 96 hours after administration. [Figure 9] 9A-9D are graphs showing COXII expression induced by administration of C. emblica extract in vitro, optionally with a nanoparticle delivery vehicle, at 6 and 24 hours post-administration. [Figure 10] 10A-10B include graphs showing expression of COXII and TFAM 6 hours after in vitro administration of components of C. emblica extract, according to one embodiment. [Figure 11] 11A-11D include graphs showing expression of COXII and TFAM 12 and 24 hours after in vitro administration of components of an extract of C. emblica, according to one embodiment. [Figure 12] 12A-12D include graphs showing expression of COXII and TFAM 12 and 24 hours after in vitro administration of components of Emblica extract, according to one embodiment. [Figure 13] 13A-13D include graphs showing expression of COXII and TFAM 12 and 24 hours after in vitro administration of components of an extract of C. emblica, according to one embodiment. [Figure 14] 14A-14D include graphs showing expression of COXII and TFAM 12 and 24 hours after in vitro administration of components of an extract of C. emblica, according to one embodiment. [Figure 15] 15A-15D include graphs showing expression of COXII 6, 12, 24, and 48 hours after in vitro administration of components of C. emblica extract, according to one embodiment. [Figure 16] 16A-16D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of components of T. emblica extract, according to one embodiment. [Figure 17]17A-17D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of components of C. emblica extract, according to one embodiment. [Figure 18] 18A-18D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of components of T. emblica extract, according to one embodiment. [Figure 19] 19A-19D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of components of C. emblica extract, according to one embodiment. [Figure 20] 20A-20D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of components of an extract of T. emblica, according to one embodiment. [Figure 21] 21A-21D include graphs showing expression of COXII 6, 12, 24, and 48 hours after in vitro administration of components of an extract of C. emblica, according to one embodiment. [Figure 22] 22A-22D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of components of an extract of T. emblica, according to one embodiment. [Diagram 23] 23A-23D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of chebulinic acid, optionally encapsulated in a nanoparticle carrier, according to one embodiment. [Figure 24] 24A-24D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of chebulinic acid, optionally encapsulated in a nanoparticle carrier, according to one embodiment. [Diagram 25] 25A-25B include graphs showing expression of COXII 6 hours after in vitro administration of chebulinic acid, optionally encapsulated in a nanoparticle carrier, according to one embodiment. [Figure 26-1]26A-26D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 26-2] 26A-26D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 27-1] 27A-27D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of T. emblica extract encapsulated in a nanoparticle carrier, according to one embodiment. [Figure 27-2] 27A-27D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of T. emblica extract encapsulated in a nanoparticle carrier, according to one embodiment. [Figure 28-1] 28A-28D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 28-2] 28A-28D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 29-1] 29A-29D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 29-2] 29A-29D include graphs showing COXII expression 6, 12, 24, and 48 hours after in vitro administration of C. emblica extract encapsulated in nanoparticle carriers, according to one embodiment. [Figure 30-1] 30A-30D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of T. emblica extract encapsulated in a nanoparticle carrier, according to one embodiment. [Figure 30-2] 30A-30D include graphs showing expression of TFAM 6, 12, 24, and 48 hours after in vitro administration of T. emblica extract encapsulated in a nanoparticle carrier, according to one embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0115] Detailed Description Mitochondrial dysfunction is associated with many mitochondrial diseases, many of which are the result of dysfunctional mitochondrial oxidative phosphorylation (OXPHOS). Mitochondrial OXPHOS constitutes the majority of cellular adenosine triphosphate (ATP) production in cells. OXPHOS function is highly dependent on the coordinated expression of proteins encoded by both the nuclear and mitochondrial genomes. The human mitochondrial genome encodes 13 polypeptides of the OXPHOS system, while the nuclear genome encodes the remaining 85+ polypeptides required for the assembly of the OXPHOS system. mtDNA depletion impairs OXPHOS, resulting in mtDNA depletion syndromes (Alberio,et al., Mitochondrion 7, 6-12, 2007;Ryan, Met al., Annu. Rev. Biochem. 76, 701-722, 2007). mtDNA depletion syndromes are a heterogeneous group of disorders characterized by low mtDNA levels in specific tissues. In different target organs, mtDNA depletion leads to specific pathological changes (Tuppen, et al., Biochim. Biophys. Acta 1797, 113-128, 201). mtDNA depletion syndromes result from genetic defects in nuclear-encoded genes involved in mtDNA replication, as well as mitochondrial nucleotide metabolism and nuclear salvage pathways (Alberio,et al., Mitochondrion 7, 6-12, 2007). mtDNA depletion is also involved in other human diseases and conditions, such as, but not limited to, mtDNA depletion syndromes, mitochondrial diseases, viral infections, conditions induced by viral infections, aging, age-related chronic diseases or conditions, reduced energy levels and vitality, hair aging features including hair loss and graying, skin aging features including skin wrinkles and senile lentigines, skin diseases and conditions, and other human pathologies.

[0116] Fertility and oocyte mitochondrial mass decrease typically occur with aging.Embryonic development and embryonic fertility may also decrease with age.In addition, older individuals are often at higher risk of ovarian disease or disorder.It is also hypothesized that fertility or oocyte mitochondrial mass decrease, embryonic development and embryonic fertility decrease, and certain ovarian diseases and disorders may occur as a result of premature aging.For example, primary ovarian insufficiency (POI) is a disorder associated with the decrease of ovarian function before the age of 40.

[0117] A general decline in mitochondrial function has been reported widely during aging, and mitochondrial dysfunction is known to be a driving force underlying age-related human diseases. Mice with specific mtDNA mutations have been shown to present signs of premature aging (i.e., mtDNA-depleted mice). In addition to mutations in mtDNA, studies have also suggested a decrease in mtDNA content and mitochondrial copy number with age. Low mtDNA copy number is associated with frailty and is a predictor of total mortality in multi-ethnic populations. A recent study revealed that humans, on average, lose approximately four copies of mtDNA every decade. This study identified an association between the decrease in mtDNA copy number and age-related physiological parameters.

[0118] Accumulating evidence suggests a strong association between mitochondrial dysfunction, mitochondrial disease, aging, and age-related diseases. In particular, increased somatic mtDNA mutations and decreased mitochondrial function have been reported widely during human aging. Studies have also suggested a decrease in mtDNA content and mitochondrial number with age.

[0119] definition As used herein, the term "carrier" refers to a diluent or vehicle with which a compound is administered. The carrier may be a pharma- ceutically acceptable carrier. The carrier may be a cosmetically acceptable carrier. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin.

[0120] As used herein, the term "corresponding aspartic acid" refers to an aspartic acid (D) residue that is mutated to an alanine (A) residue in the POLG1 amino acid sequence that is equivalent to the aspartic acid at position 1135 of the human POLG1 sequence. In certain embodiments, the "corresponding aspartic acid" is flanked on the amino-terminal side by an amino acid sequence of S / TI / VHX, IS / TI / VHX, or C / AIS / TI / VHX, and / or on the carboxy-terminal side by an amino acid sequence of XEV / IR, XEV / IRY / F, or XEV / IRY / FL, where "X" indicates the aspartic acid amino acid that is or will be mutated to alanine.

[0121] As used herein, the term "depleted", "depleted" or "depleter" with respect to mtDNA refers to a reduction in copy number and / or concentration of mtDNA in a human or non-human animal, tissue or cell of a non-human animal. Such determinations can be made with respect to a human not administered a compound or composition of the present disclosure, or a control non-human animal tissue or cell (e.g., a non-human animal that does not express or does not express a mutant POLG1 polypeptide).

[0122] As used herein, "treatment" of a disease or condition refers to reducing or decreasing the severity or frequency of at least one symptom of the disease or condition, compared to a similar but untreated patient. Treatment can also refer to stopping, slowing, or reversing the progression of a disease or condition, compared to a similar but untreated patient. Treatment can include addressing the underlying cause of the disease and / or one or more symptoms.

[0123] As used herein, the term "effective amount" refers to an amount of a compound or composition of the present disclosure administered to a subject that is effective to provide a desired response and / or effect in the subject. The response and / or effect may be a cosmetic response and / or effect. The response and / or effect may be a therapeutic response and / or effect. The response and / or effect may be a prophylactic or preventative response and / or effect.

[0124] As used herein, the term "therapeutically effective amount" refers to an amount of a compound or composition of the present disclosure administered to a subject that is effective to treat a disease or condition described herein in the subject and / or to produce a desired physiological response and / or therapeutic effect in the subject. One example of a desired physiological response includes increasing the copy number and / or concentration of mtDNA.

[0125] The actual dose, including an "effective amount" or a "therapeutically effective amount", can depend on the route of administration, the size and health of the subject, the disorder being treated, and the like.

[0126] In some embodiments, an "effective amount" or "therapeutically effective amount" in the context of this disclosure may be sufficient to treat or prevent an ovarian disease or condition, and its symptoms.

[0127] In some embodiments, an "effective amount" or "therapeutically effective amount" in the context of the present disclosure is sufficient to induce mitochondrial biogenesis. An "effective amount" or "therapeutically effective amount" may be sufficient to induce mitochondrial biogenesis locally. An "effective amount" or "therapeutically effective amount" may be sufficient to induce mitochondrial biogenesis systemically.

[0128] In some embodiments, an "effective amount" or a "therapeutically effective amount" in the context of the present disclosure is an amount that reduces an inflammatory phenotype, increases expression of a mitochondrial oxidative phosphorylation complex, increases the stability of a mitochondrial oxidative phosphorylation complex, alters, e.g., decreases, the expression of at least one gene selected from the group consisting of NF-κB, COX-2, INF-β1, CCL5, MMP1, MMP2, MMP9, MMP13, IGF1R, VEGF, and MRPS5, and increases, e.g., decreases, the expression of at least one gene selected from the group consisting of TIMP1. Alter, e.g., increase, the expression of at least one gene selected from the group consisting of KLOTHO, COL1A1, MTCO2, TFAM, and VDAC; prevent deactivation of genes related to mitochondrial health and activity selected from FGF2, FGFR1, COX7A1, PDK4, FAM173A, MRPL12, and WNT11; reduce inflammatory infiltrates in the skin and / or hair follicles; and / or increase the expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COXII.

[0129] In some embodiments, an "effective amount" or "therapeutically effective amount" in the context of the present disclosure increases mtDNA copy number or concentration by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%. In each of the foregoing, where a reduction or increase is defined, such reduction or increase may be determined with respect to a subject or non-human animal that has not been treated with a compound or composition of the present disclosure and that is suffering from a disease or condition described herein.

[0130] In some embodiments, an "effective amount" or a "therapeutically effective amount" in the context of the present disclosure is an amount that prevents the development of an inflammatory phenotype, prevents a decrease in expression of mitochondrial oxidative phosphorylation complexes, prevents a decrease in stability of mitochondrial oxidative phosphorylation complexes, prevents an increase in expression of at least one gene selected from the group consisting of NF-κB, COX-2, INF-β1, CCL5, MMP1, MMP2, MMP9, MMP13, IGF1R, VEGF, and MRPS5, prevents an increase in expression of at least one gene selected from the group consisting of TIMP1, TIMP2, TIMP3, TIMP4, TIMP5, TIMP6, TIMP7, TIMP8, TIMP9, TIMP10, TIMP11, TIMP12, TIMP13, TIMP14, TIMP15, TIMP16, TIMP17, TIMP18, TIMP19, TIMP20, TIMP21, TIMP22, TIMP23, TIMP24, TIMP25, TIMP26, TIMP27, TIMP28, TIMP29 ...8, TIMP29, TIMP29, TIMP20, TIMP21, TIMP22, TIMP23, TIMP24, TIMP25, TIMP25, TIMP26, TIMP27, TIMP28, TIMP29, TIMP29, TIMP28, TIMP29, Prevents decreased expression of at least one gene selected from the group consisting of KLOTHO, COL1A1, MTCO2, TFAM, and VDAC; prevents inactivation of genes related to mitochondrial health and activity selected from FGF2, FGFR1, COX7A1, PDK4, FAM173A, MRPL12, and WNT11; prevents increased inflammatory infiltrates in the skin and / or hair follicles; prevents decreased copy number or concentration of mtDNA; and / or prevents decreased expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COXII.

[0131] As used herein, the term "excipient" refers to a substance formulated with the active ingredient of the composition, including, but not limited to, for purposes such as long-term stabilization, bulking up solid formulations containing small amounts of potent active ingredients (hence often referred to as bulking agents, fillers, or diluents), or to impart therapeutic enhancements to the active ingredient in the final dosage form, such as facilitating drug absorption, reducing viscosity, or enhancing solubility. Excipients may also be useful in the manufacturing process to aid in in vitro stability, such as preventing denaturation or aggregation over the expected shelf life, as well as to aid in concerns in handling the active, such as by promoting powder flowability or non-stickiness. The excipient may be a pharma- ceutically acceptable excipient. The excipient may be a cosmetically acceptable excipient. Examples of suitable excipients are described in "Remington's Pharmaceutical Sciences" by E. W. Martin.

[0132] As used herein, the term "in need of" (such as in the phrase "in need of treatment") refers to a judgment made by a medical professional that a subject requires or would benefit from administration of a compound of the present disclosure. This judgment is made based on a variety of factors within the medical professional's area of ​​expertise, including, but not limited to, knowledge that the subject is ill or will become ill as a result of a disease or condition treatable by a method or pharmaceutical composition of the present disclosure.

[0133] As used herein, the term "mutated POLG1" or "mutated POLG1" refers to a POLG1 amino acid sequence from a particular species that contains at least one mutation compared to a wild-type POLG1 sequence from that species. The mutation does not have to cause disease. There may be a single mutation or two or more mutations. In certain embodiments, a single dominant negative mutation may be present, optionally with one or additional mutations (such as, but not limited to, the D1135A mutation).

[0134] As used herein, the term "pharmaceutical acceptable" refers to a compound that is compatible with other components of a composition and is not harmful to the subject receiving the compound or composition. In some embodiments, the term "pharmaceutical acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals, more particularly humans.

[0135] As used herein, the term "cosmetically acceptable" refers to a compound that is compatible with other cosmetic ingredients of a composition and is not harmful to the subject receiving the compound or composition. A cosmetically acceptable composition or compound may be pharmacologic acceptable. However, a cosmetically acceptable composition or compound does not have to be pharmacologic acceptable.

[0136] As used herein, the term "pharmaceutically acceptable form" is meant to include known forms of the compounds of the present disclosure that may be administered to a subject, including, but not limited to, solvates, hydrates, prodrugs, isomorphs, polymorphs, pseudomorphs, neutral forms, and salt forms of the compounds of the present disclosure.

[0137] As used herein, the term "pharmaceutically acceptable salts" refers to physiologically and pharma- ceutically acceptable salts of compounds of the present invention, i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects of the disclosed compounds. For oligonucleotides, exemplary pharma- ceutically acceptable salts include, but are not limited to, (a) salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamines such as spermine and spermidine, and the like; (b) acid addition salts formed with inorganic acids, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like; (c) salts formed with organic acids, e.g., acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (d) salts formed from elemental anions such as chlorine, bromine, and iodine.

[0138] As used herein, the term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure with other components, such as, but not limited to, pharma- ceutically acceptable carriers and / or excipients. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure.

[0139] In some embodiments, a "cosmetically acceptable" excipient refers to a cosmetically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. In some embodiments, each excipient is cosmetically acceptable in the sense that it is compatible with other ingredients of a cosmetic formulation and is suitable for use in contact with human and animal tissues or organs without undue toxicity, irritation, allergic reaction, immunogenicity, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0140] As used herein, the term "replete," "repletion," or "repletion agent" with respect to mtDNA refers to an increase in copy number and / or concentration of mtDNA in a human or non-human animal, tissue, or cell. Such determination can be performed with respect to a human or control non-human animal tissue, or cell that has undergone mtDNA depletion (such as immediately before repletion is initiated). In certain embodiments, repletion of mtDNA results in a copy number and / or concentration of mtDNA that is approximately equal to the copy number and / or concentration of mtDNA observed in a control non-human animal, tissue, or cell (e.g., a non-human animal that does not express or does not express a mutant POLG1 polypeptide).

[0141] As used herein, the term "solvate" refers to a compound of the present disclosure, or a pharma- ceutically acceptable salt thereof, in which one or more molecules of a suitable solvent are incorporated into the crystal lattice. A suitable solvent is physiologically tolerable at the dosage administered. Examples of suitable solvents are ethanol, water, etc. When water is the solvent, the molecule may be referred to as a "hydrate."

[0142] As used herein, the term "subject" or "patient" includes all members of the animal kingdom, including, but not limited to, vertebrates, mammals, animals (e.g., cats, dogs, horses, pigs, rodents, etc.), and humans. In certain embodiments, the subject is a human. In certain embodiments, the subject is an animal, e.g., a research animal, e.g., a mouse, a rat.

[0143] As used herein, the term "similarity" or "similarity score", when used in reference to a compound, refers to the degree of similarity between two or more compounds in chemical structure, chemical function, and / or one or more chemical properties. The similarity score can be quantified by augmenting data generated by a deep neural network trained to model multiple chemical reactions for any given compound. The data can be augmented by multi-dimensional vectors that define a matrix of the compound's properties or chemical reactions in which the compound participates to generate an embedding score for the compound. The embedding scores of two or more compounds can be compared to generate a similarity score between two or more compounds.

[0144] All patent applications, patents, and printed publications cited herein are hereby incorporated by reference in their entirety for all purposes.

[0145] The impact of mitochondrial function on intrinsic and extrinsic aging Mitochondrial dysfunction affects both intrinsic and extrinsic aging. Skin wrinkling, acanthosis, epidermal hyperplasia with hyperkeratosis, and the presence of significant inflammatory infiltrates in the skin have been observed in mtDNA-depleted mice, representing features similar to extrinsic aging of the skin in humans. Furthermore, changes in the expression of intrinsic aging-related gene markers support an intrinsic mechanism underlying the phenotypic changes observed in mtDNA-depleted mice.

[0146] It has been reported that loss of collagen fibers is the basis of skin wrinkling. A tight balance between proteolytic matrix metalloproteinase (MMP) enzymes and their tissue-specific inhibitor tissue inhibitor of metalloproteinase-1 (TIMP1) is essential to maintain collagen fiber content in skin. Expression of MMPs is altered in aged skin. Consistent with these reports, skin of mtDNA-depleted mice showed increased expression of MMPs and decreased expression of TIMP1, indicating a loss of balance that contributes to the development of skin wrinkles. Repletion of mtDNA content restored MMP expression leading to wrinkled skin and reversal of hair loss. These experiments indicate that mitochondria are regulators of aging. This observation is surprising and suggests that epigenetic mechanisms underlying mitochondrial-nuclear crosstalk must play an important role in the restoration of normal skin and hair phenotypes.

[0147] mtDNA stress induces an inflammatory response. Inflammation also underlies aging and age-related diseases. Increased levels of markers of inflammation in mtDNA-depleted mice indicate an activated immune response in the skin of mtDNA-depleted mice. The increased expression of NF-κB, a master regulator of inflammatory responses upon mtDNA depletion, and its reduced expression after restoration of mtDNA content, suggest that NF-κB signaling is an important mechanism contributing to the skin and hair follicle phenotype observed in mtDNA-depleted mice. Furthermore, a unique property of proteins encoded by mtDNA is the N-terminus N-formyl-methionine. N-formylated peptides, when present in the extracellular space, are known to act as mitochondrial damage-associated molecular patterns and to activate neutrophils or keratinocyte intrinsic responses leading to the recruitment of immune cells. While previous animal models have used localized approaches (e.g., targeting only specific cell types) to show alterations in mtDNA homeostasis and / or copy number or concentration of mtDNA, the present disclosure utilizes an animal model that provides global and targeted disruption of mtDNA homeostasis and / or copy number or concentration of mtDNA in a controlled manner. Using such an animal model, the present disclosure has identified compounds that are effective in treating diseases and conditions related to mitochondrial dysfunction in the background of significant mtDNA depletion. In addition, the present disclosure demonstrates that such compounds reverse the physiological and phenotypic effects mediated by mitochondrial dysfunction. Exemplary physiological and phenotypic effects of intrinsic and extrinsic aging that are reversed include reduced skin wrinkles, reduced hair loss, increased hair follicles in anagen, reduced inflammatory gene expression, reduced inflammatory infiltrate in skin and hair follicles, and increased collagen content in skin.

[0148] In addition, due to the short life span of conventional animal models, previous studies have not been able to determine the effect of mitochondrial dysfunction on aging process, and such studies have not observed the appearance of many of the physiological and phenotypic changes related to mitochondrial dysfunction.Therefore, conventional practices have not been able to identify the compounds that are effective in treating such physiological and phenotypic changes related to mitochondrial dysfunction reported herein.

[0149] The above limitations identify effective treatments for skin diseases and conditions where mitochondrial dysfunction is an unsolved challenge. In addition, due to abnormalities in normal epidermal development and hair follicle morphogenesis in current models, false positive and false negative results are common. The present disclosure utilizes an inducible non-human animal model that expresses mutated POLG1 polypeptides (such as, but not limited to, POLG1 polypeptides expressing dominant negative (DN) mutations) that induce mitochondrial dysfunction (e.g., by depletion of mtDNA) in whole animals or selected cells / tissues. The non-human animal model allows for the ubiquitous suppression and restoration of mitochondrial function in whole animals or specific cells / tissues.

[0150] The disclosed non-human animal model can be used to rapidly identify compounds that are effective in treating diseases and conditions related to mtDNA.The animal model in the absence of expression of mutant POLG1 expression maintains normal epidermal differentiation and hair follicle morphogenesis, making this animal model system a valuable tool in identifying treatments for various diseases and conditions characterized by mitochondrial dysfunction.When mutant POLG1 polypeptide is expressed, the animal model demonstrates significant phenotypic age-related changes in skin, including the development of skin wrinkles and hair loss.Phenotypic changes are reversible when mitochondrial function is restored (such as by administration of the extract or compound described herein).

[0151] Non-human animal models In certain aspects, the methods described herein utilize genetically modified non-human animals that express mutant POLG1 polypeptides in a controlled manner. Tissues, organs and cells from such animal models can also be used. In one embodiment, mutant POLG1 polypeptides are ubiquitously expressed (in every cell of the non-human animal). In another embodiment, mutant POLG1 polypeptides are expressed in a specific tissue or set of tissues, or in a specific cell type. Such non-human animal models are described in U.S. Patent Publication No. 2020-0085021-A1, which is incorporated herein by reference in its entirety for all purposes.

[0152] In certain embodiments, the animal model exhibits at least one feature selected from the group consisting of reduced mitochondrial (mt) DNA content, reduced mtDNA copy number, altered mitochondrial protein expression, reduced expression of mitochondrial oxidative phosphorylation complexes, reduced stability of mitochondrial oxidative phosphorylation complexes, skin wrinkling, hair loss, increased epidermal thickness, epidermal hyperplasia, acanthosis, hyperkeratosis, altered (e.g., increased) expression of at least one gene selected from the group consisting of NF-κB, COX-2, INF-β1, CCL5, MMP1, MMP2, MMP9, MMP13, IGF1R, VEGF, and MRPS5, altered (e.g., decreased) expression of at least one gene selected from the group consisting of TIMP1 and KLOTHO, COL1A1, MTCO2, TFAM, and VDAC, increased skin inflammation, and abnormal hair follicles.

[0153] Methods for screening for therapeutic agents The present disclosure provides an artificial neural network trained to model multiple chemical reactions for any given compound. By using the artificial neural network described herein, compounds and compositions that have a similarity score with known promoters or inhibitors of mitochondrial biogenesis can be easily identified. The similarity score can be quantified by expanding the data generated by the deep neural network in a multi-dimensional vector that defines a matrix of the compound's properties or chemical reactions in which the compound participates to create an embedding score for the compound. The embedding score for the compound can be compared to the embedding scores for known promoters or inhibitors of mitochondrial biogenesis to create a similarity score. Thus, the present disclosure provides a method for screening compounds and compositions that have sufficient similarity scores with known compounds. The identified compounds are predicted to be effective in promoting or inhibiting mitochondrial biogenesis.

[0154] The present disclosure provides a non-human animal model that expresses mutant POLG1 polypeptide in a controlled manner, either throughout the animal or in specific tissues, including mutant POLG1 polypeptide.By using the non-human animal model described herein, the compound and composition that are effective in treating diseases and conditions related to mitochondrial dysfunction can be easily identified.Methods that utilize cells and tissues from such non-human animals are also provided.Therefore, the present disclosure provides a method for screening the compound and composition that are effective for treating diseases and conditions related to mitochondrial dysfunction in subjects, including diseases and conditions related to mtDNA depletion.

[0155] In one embodiment, the present disclosure provides for the identification of compounds for the treatment of diseases or conditions that are at least partially caused by mitochondrial dysfunction, including changes in mtDNA copy number and / or concentration and / or dysfunctional mitochondrial OXPHOS.Such diseases and conditions include, but are not limited to, mtDNA depletion syndrome, mitochondrial diseases, ovarian diseases or conditions and their symptoms, aging, age-related chronic diseases, reduced energy levels and vitality, and other human pathologies.Exemplary mitochondrial diseases include cardiovascular disease, diabetes, cancer, neuropathy, such as age-related neuropathy, skin diseases and conditions, such as skin wrinkles, skin pigmentation changes, senile lentigines, skin aging features, hair or scalp diseases and conditions, such as hair loss, thinning hair, hair pigmentation changes, such as hair graying.

[0156] In one embodiment, such a screening method includes the steps of: a) providing a deep neural network trained to model multiple chemical reactions for known promoters or inhibitors of mitochondrial biogenesis; b) running the deep neural network to identify one or more compounds having a similarity score threshold of at least 70% with known promoters or inhibitors of mitochondrial biogenesis; c) selecting one or more identified compounds based on at least one inclusion criterion; and d) evaluating at least one selected compound in an assay to determine the effect of the selected compound on mitochondrial biogenesis, where the assay may include an in vitro assay, an ex vivo assay, or an in vivo assay. The at least one inclusion criterion may include structural, functional, physical, or chemical properties of the identified compound. Exemplary properties include size, charge, ionic strength, binding strength, valency, hybridization, micromolecular structure, macromolecular structure, and the like. The similarity score threshold may be at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, at least 99.5%, at least 99.9%, at least 99.95%, at least 99.99%, at least 99.995%, at least 99.999%, or higher.

[0157] In some embodiments, the assay may include screening at least one selected compound using a non-human animal capable of inducible expression of a mutant POLG1 polypeptide as described herein. In some embodiments, the assay may include screening at least one selected compound to measure an effect on protein or gene expression of a mitochondrial health biomarker as specified herein. The screening method may be repeated by running a deep neural network to identify one or more compounds having a similarity score of at least 70% with the at least one identified compound and / or the at least one selected compound. In some embodiments, the deep neural network may be retrained with the at least one identified compound and / or the at least one selected compound.

[0158] In one embodiment, such a screening method comprises the steps of: a) providing a non-human animal capable of inducible expression of a mutant POLG1 polypeptide; b) stimulating expression of the mutant POLG1 polypeptide, whereby stimulating expression of the mutant POLG1 polypeptide induces a physiological or phenotypic response; c) administering an agent to the non-human animal either before step b) or after step b); d) determining the effect of the agent on the pathology; and e) comparing the effect of the agent with a control animal, whereby a reduction or increase (if appropriate) in the physiological or phenotypic response in the non-human animal after administration of the agent indicates that the agent is a therapeutic agent for treating the physiological or phenotypic response.

[0159] In another embodiment, the disclosure provides a method for identifying a therapeutic agent for the treatment of mitochondrial dysfunction.

[0160] In another embodiment, the disclosure provides a method for identifying a therapeutic agent for the treatment of a disease or condition associated with mitochondrial dysfunction, or a symptom thereof.

[0161] In another embodiment, the disclosure provides a method for identifying a therapeutic agent for the treatment of an age-related chronic disease related to mitochondrial dysfunction, or a symptom thereof.

[0162] In another embodiment, the disclosure provides methods for identifying therapeutic agents for improving or preventing reduced energy levels and / or vitality or conditions.

[0163] In another embodiment, the present disclosure provides a method for identifying therapeutic agents for treating or preventing ovarian disease or condition and its symptoms.In any of the described screening methods, the agents may include, but are not limited to, chemical compounds, pharmaceutical compositions, cosmetic compositions, extracts, plant extracts, seaweed extracts, microbial extracts, biological compounds and compositions (e.g., proteins, DNA, RNA, siRNA, vaccines, etc.), and microorganisms.Furthermore, the agents may be selected from libraries, including libraries of agents approved by regulatory agencies such as the FDA.

[0164] Any of the transgenic non-human animals of the present disclosure may be used in any of the screening methods described.

[0165] In any of the screening methods described, step b) may be achieved by providing an inducer compound to the transgenic non-human animal or by withholding the inducer compound from the transgenic non-human animal.

[0166] In any of the screening methods described, the agent is added before step b). In any of the screening methods described, the agent is added after step b).

[0167] In any of the described screening methods, the animal model is the animal model described in the preceding section.In any of the described screening methods, the mutant POLG1 polypeptide can be any mutant POLG1 polypeptide described herein.In certain embodiments, the mutant POLG1 polypeptide comprises a dominant negative mutation.In certain embodiments, the mutant POLG1 polypeptide comprises D1135A mutation.

[0168] Treatment Method In one embodiment, the present disclosure provides a method for treating or preventing an age-related chronic disease or condition related to mitochondrial dysfunction in a subject, or a symptom thereof, comprising administering to the subject an effective amount of a compound of the present disclosure, or a pharma- ceutically acceptable form thereof. In one embodiment, the present disclosure provides a method for treating or preventing an age-related chronic disease or condition related to mitochondrial dysfunction in a subject, or a symptom thereof, comprising administering to the subject an effective amount of a compound of the present disclosure, or a pharma- ceutically acceptable form thereof.

[0169] The present disclosure provides compounds and compositions that are effective for treating or preventing diseases and conditions related to mitochondrial dysfunction, including mtDNA depletion. Diseases and conditions related to mitochondrial dysfunction include ovarian diseases or conditions and their symptoms. Ovarian diseases or conditions that can be treated by the methods disclosed herein include, for example, perimenopause, menopause, endometriosis, ovarian cysts, premenopausal or postmenopausal ovarian cancer, such as ovarian epithelial cancer, ovarian tumors, such as ovarian germ cell tumors, ovarian low malignant potential tumors, and ovarian stromal tumors, polycystic ovarian syndrome (PCOS), primary ovarian insufficiency (POI), and ovarian torsion. Ovarian diseases or conditions can be associated with infertility, premature ovarian aging, and reduced mitochondrial mass of oocytes. Treatment can include, for example, reducing or decreasing the severity or frequency of symptoms of ovarian diseases or conditions, including dry skin or vagina, pelvic pain or muscle spasms, inflammation, extended or irregular menstrual cycles, and reduced ovulatory events. Treatment or prevention may involve inducing mitochondrial biogenesis and / or improving mitochondrial function.

[0170] Thus, in certain embodiments, the present disclosure provides compounds and compositions effective for treating or preventing, e.g., reducing, inhibiting, delaying, and / or reversing perimenopause, menopause, and / or symptoms thereof. Perimenopause is the stage before menopause, or the final period. Perimenopause may be characterized by changes in hormone levels, e.g., reduced estrogen and / or increased follicle-stimulating hormone (FSH), reduced ovulation, and extended or irregular menstrual cycles. In certain examples, perimenopause may be characterized by FSH levels of about 20 IU / mL to 30 IU / mL or higher in blood samples, and menopause may be characterized by FSH levels of about 25 IU / mL to 135 IU / mL or higher in blood samples. However, in general, any changes in hormone levels may be compared to the subject's baseline levels to determine whether the subject is experiencing perimenopause or postmenopause. Symptoms of perimenopause and menopause can include mood changes, changes in libido, trouble concentrating, headaches, hot flashes, night sweats, vaginal dryness, trouble sleeping, joint or muscle pain, heavy sweating, frequent urination, and other premenstrual (PMS)-like symptoms.

[0171] In certain embodiments, ovarian disease or condition and its symptoms include those related to mitochondrial dysfunction.Without being bound by any particular theory, the compound disclosed herein may show the observed effect by inhibiting the decrease of mitochondrial DNA copy number or concentration, inhibiting the depletion of mitochondrial DNA, inhibiting the degradation of mitochondrial DNA, contributing to the increase of mitochondrial DNA copy number or concentration, replenishing mitochondrial DNA, and / or promoting the increase of mitochondrial DNA activity.

[0172] The method disclosed herein relates to enhancing or improving fertility in a subject.Any reduction in fertility of a subject can be age-related or premature.For example, the disease, condition or symptom disclosed herein can be associated with age-related ovarian aging or premature ovarian aging.The method disclosed herein relates to extending the reproductive lifespan of a subject.

[0173] According to certain embodiments, the methods disclosed herein relate to increasing the mitochondrial mass of an oocyte in a subject or preventing a decrease in the mitochondrial mass of an oocyte.

[0174] Certain extracts have been identified as containing one or more compounds that promote and / or inhibit mtDNA. emblica extract, fucus extract, and chebula extract are described herein. It should be understood that similar extracts, particularly extracts containing one or more compounds disclosed herein, compounds that constitute the extracts disclosed herein, and / or compounds that have at least a 95% similarity score with one or more compounds disclosed herein, are expected to provide similar mtDNA promotion and / or inhibition. Thus, other extracts, compounds derived from, constitute, or purified from other extracts, and compounds that have at least a 95% similarity score with compounds derived from, constitute, or purified from other extracts are within the scope of the present disclosure.

[0175] Exemplary extracts include Polygonum aviculare extract, Physalis gngulata extract, Dunaliella salina extract, Camellia sinensis leaf extract, Tremella fuciformis sporocarp extract, Alteromonas ferment extract, Theobroma cacao (cocoa) seed extract, Vitis vinifera (grape) flower cell extract, Mirabilis jalapa callus extract, Alteromonas ferment extract, and Vibrio alginolyticus ferment filtrate.

[0176] Thus, the compounds described herein may be derived from, purified from, or isolated from the extract. The compounds described herein may be derived from, purified from, or isolated from a source other than the extract. The compound components of the extract may be derived from, purified from, or isolated from another natural or artificial source. In other embodiments, the compounds described herein may be synthetic. For example, the compounds of the present disclosure may be synthesized in a laboratory, in a manufacturing, or in other situations. The above applies to compounds contained in the extract or its components, and to compounds with high similarity scores.

[0177] The methods disclosed herein may comprise administering to the subject an effective amount of an emblica extract or a pharma-ceutically acceptable form thereof.

[0178] The methods disclosed herein may include administering to the subject an effective amount of a compound derived from, consisting of, or purified from an emblica extract, or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% with a compound derived from, consisting of, or purified from an emblica extract, or a pharma- ceutically acceptable form thereof.

[0179] The methods disclosed herein may include administering to the subject an effective amount of a fucus extract or a pharma- ceutically acceptable form thereof.

[0180] The methods disclosed herein may include administering to the subject an effective amount of a compound derived from, constituting, or purified from a fucus extract, or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% to a compound derived from, constituting, or purified from a fucus extract, or a pharma- ceutically acceptable form thereof.

[0181] The methods disclosed herein can include administering to the subject an effective amount of a chebula extract or a pharma- ceutically acceptable form thereof.

[0182] The methods disclosed herein may include administering to the subject an effective amount of a compound derived from, consisting of, or purified from a chebula extract, or a pharma- ceutically acceptable form thereof, or a compound having a similarity score of at least 95% with a compound derived from, consisting of, or purified from a chebula extract, or a pharma- ceutically acceptable form thereof.

[0183] The present disclosure may generally relate to extracts and compounds derived from, constitute or purified from extracts.It should be understood that compounds having at least 95%, for example at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.9%, at least 99.95%, at least 99.99%, at least 99.995%, or at least 99.999% similarity score with compounds derived from, constitute or purified from extracts can be utilized in any composition described herein.

[0184] Without wishing to be bound by theory, it is believed that the administration of the compositions disclosed herein can be involved in inhibiting the decrease in mitochondrial DNA copy number or concentration, inhibiting mitochondrial DNA depletion, inhibiting mitochondrial DNA degradation, or a combination thereof. Without wishing to be bound by theory, it is believed that the administration of the compositions disclosed herein can be involved in inducing mitochondrial biogenesis and / or improving mitochondrial function.

[0185] According to certain embodiments, administration of the compositions disclosed herein may involve increased expression of mitochondrial oxidative phosphorylation complexes, increased stability of mitochondrial oxidative phosphorylation complexes, and / or increased expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COXII.

[0186] According to certain embodiments, administering the compositions disclosed herein may include modulating menstrual cycle, for example, normalizing menstrual cycle, for example, regulating the amount and / or frequency of menstrual cycle.Administering the compositions disclosed herein may involve increasing ovulation events.Administering the compositions disclosed herein may involve increasing the anti-Mullerian hormone (AMH) level of the subject.Furthermore, administering the compositions disclosed herein may reduce the probability of embryonic aneuploidy and / or Leigh syndrome.

[0187] In certain aspects, the emblica extract is derived from Emblica officinalis (also known as Phyllanthus emblica, Indian gooseberry, or amla by its Hindi name). Such extracts from Emblica officinalis may be derived from any part of the plant, as desired. For example, the extract may be derived from the stem, fruit, or both the stem and fruit of Emblica officinalis. In preparing such extracts, Emblica officinalis may be provided in a powdered form and extracted using chemical solvents known in the art, such as, but not limited to, aqueous ethyl acetate, ethanol, and methanol. Other solvents or excipients disclosed herein may be used. In certain exemplary embodiments, the chemical solvent may be methanol.

[0188] In certain embodiments, an effective amount of emblica extract, or a compound derived from, constituting, or purified from emblica extract, is between 1 and 100 mg, e.g., 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg.

[0189] In certain embodiments, the compounds derived from, constituting, or purified from an emblica extract are degradation products of tannins, where the emblica extract is as described above, as appropriate. In certain embodiments, the compounds derived from, constituting, or purified from an emblica extract are ellagitannins. In certain embodiments, the compounds derived from, constituting, or purified from an emblica extract are emblicanin A, emblicanin B, punigluconin, pedunculagin, and / or chebulinic acid.

[0190] In certain embodiments, the compound is an ellagitannin or a compound having at least a 95% similarity score with an ellagitannin. Exemplary ellagitannins and compounds having high similarity scores with such ellagitannins are listed in Table 1. Other compounds having high similarity scores with ellagitannins are within the scope of the present disclosure. Table 1: Ellagitannins and compounds with high similarity scores to ellagitannins [Table 1-1] [Table 1-2] [Table 1-3]

[0191] In certain embodiments, the compound is chebulinic acid or a compound having at least 95% similarity score with chebulinic acid (Formula I below). Exemplary compounds having high similarity score with chebulinic acid are listed in Table 2.

[0192] Table 2: Compounds with high similarity scores to chebulinic acid. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8]

[0193] In certain embodiments, the compound derived from, constituting, or purified from an emblica extract is benzoic acid substituted with 1-5 hydroxy groups and, optionally, 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups. In certain embodiments, the compound derived from, constituting, or purified from an emblica extract is benzoic acid substituted with 1-3 hydroxy groups and, optionally, 1-2 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups.

[0194] In certain embodiments, the compound derived from, constituting, or purified from the extract of C. emblica is -CH=CH-(CH2) a In certain embodiments, the compound derived from, constituting, or purified from an extract of C. emblica is -CH=CH-(CH2) a In certain embodiments, the compound derived from, constituting, or purified from an extract of C. emblica is -CH=CH-(CH2) a -C(O)OH (wherein a is 0) and benzene substituted with 1 to 3 hydroxy groups.

[0195] In certain embodiments, the compounds derived from, contained in, or purified from the emblica extract are gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, emblicanin A, emblicanin B, punigluconin, and pedunculagin, punicaforin, filanembulin, kaempferol, ellagic acid, chebulinic acid, chebulaginic acid, punicalagin, or a metabolite of any of the foregoing.

[0196] In certain embodiments, the compound derived from, constituting, or purified from an extract of Emblica is ascorbic acid or citric acid.

[0197] In certain embodiments, the compound derived from, constituting, or purified from the C. emblica extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, vitamin C, or a metabolite of any of the foregoing.

[0198] In certain embodiments, the compound derived from, constituting, or purified from the C. emblica extract is gallic acid or a compound having a similarity score of at least 95% with gallic acid.

[0199] In certain embodiments, the active agent in the composition is gallic acid.The active agent may be or include ellagitannin.The active agent may be or include emblicanin A, emblicanin B, punigluconin, pedunculagin, and / or chebulinic acid.

[0200] According to certain embodiments, the composition may include an emblica extract enriched with one or more compounds that are components of the emblica extract.The compound components of the emblica extract or the combination of compound components of the emblica extract may be purified, for example, at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0201] In certain embodiments, the effective amount of a compound derived from, constituting, or purified from an extract of C. emblica, and optionally an effective amount of an active agent of the composition, is 1 to 100 mg, e.g., 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of a compound derived from, constituting, or purified from an extract of C. emblica.

[0202] In certain aspects, the fucus extract is derived from Fucus vesiculosus, Fucus serratus, Fucus spiralis, or Fucus guiryi. In certain aspects, the fucus extract is derived from Fucus vesiculosus. Such extracts may be derived from any part of the algae, as desired. In preparing such extracts, Fucus vesiculosus, Fucus serratus, Fucus spiralis, or Fucus guiryi may be provided in powdered form and extracted using chemical solvents known in the art, such as, but not limited to, aqueous ethyl acetate, ethanol, and methanol. Other solvents or excipients disclosed herein may be used. In certain exemplary embodiments, the chemical solvent may be methanol.

[0203] In certain embodiments, an effective amount of fucus extract is between 1 and 100 mg of extract, for example, 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of fucus extract.

[0204] In certain embodiments, the compound derived from, constituting, or purified from a fucus extract is benzoic acid substituted with 1-5 hydroxy groups and, optionally, 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups. In certain embodiments, the compound derived from, constituting, or purified from a fucus extract is benzoic acid substituted with 1-3 hydroxy groups and, optionally, 1-2 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups.

[0205] In certain embodiments, the compound derived from, constituting, or purified from a Fucus extract is -CH=CH-(CH2) a In certain embodiments, the compound derived from, constituting, or purified from a Fucus extract is -CH=CH-(CH2) a In certain embodiments, the compound derived from, constituting, or purified from a Fucus extract is -CH=CH-(CH2) a -C(O)OH (wherein a is 0) and benzene substituted with 1 to 3 hydroxy groups.

[0206] In certain embodiments, the compound derived from, constituting, or purified from a fucus extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, or a metabolite of any of the foregoing.

[0207] In certain embodiments, the compound derived from, constituting, or purified from a F. fucus extract is gallic acid.

[0208] In certain embodiments, the active agent in the composition is gallic acid or a compound with at least 95% similarity score with gallic acid.In certain embodiments, the active agent in the composition is chebulinic acid or a compound with at least 95% similarity score with chebulinic acid (e.g., see Table 2 above).The active agent may be or include ellagitannin or a compound with at least 95% similarity score with ellagitannin (e.g., see Table 1 above).The active agent may be or include fucoidan or a compound with at least 95% similarity score with fucoidan.

[0209] According to certain embodiments, the composition may comprise a fucus extract enriched with one or more compounds that are components of the fucus extract.The compound components of the fucus extract or the combination of compound components of the fucus extract may be purified, for example, at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0210] In certain embodiments, the effective amount of a compound derived from, constituting, or purified from a fucus extract, and optionally an effective amount of an active agent of the composition, is 1 to 100 mg, e.g., 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of a compound derived from, constituting, or purified from a fucus extract.

[0211] In certain aspects, the chebula extract is derived from Terminilia chebula, Terminalia arborea, or Lumnitzera racemose. In certain aspects, the chebula extract is derived from Terminilia chebula. Such extracts may be derived from any part of the plant, as desired. In preparing such extracts, Terminilia chebula, Terminalia arborea, or Lumnitzera racemose may be provided in powdered form and extracted using chemical solvents known in the art, such as, but not limited to, aqueous ethyl acetate, ethanol, and methanol. Other solvents or excipients disclosed herein may be used. In certain exemplary embodiments, the chemical solvent may be methanol.

[0212] In certain embodiments, an effective amount of chebula extract is between 1 and 100 mg of extract, for example, 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of chebula extract.

[0213] In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is a degradation product of tannins. In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is an ellagitannin. In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is chebulinic acid.

[0214] In certain embodiments, the compound derived from, constituting, or purified from a chebula extract is a degradation product of tannin. In certain embodiments, the compound derived from, constituting, or purified from a fucus extract is an ellagitannin. In certain embodiments, the compound derived from, constituting, or purified from a fucus extract is a fucoidan.

[0215] In certain embodiments, the compound derived from, contained in, or purified from a chebula extract is benzoic acid substituted with 1-5 hydroxy groups and optionally 1-3 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups. In certain embodiments, the compound derived from, contained in, or purified from a fucus extract is benzoic acid substituted with 1-3 hydroxy groups and optionally 1-2 O-(C1-C5 alkyl) or O-(C1-C5 alkenyl) groups.

[0216] In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is -CH=CH-(CH2) a -C(O)OH, where a is 0-5, and benzene substituted with 1-5 hydroxy groups. In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is -CH=CH-(CH2) a -C(O)OH, where a is 0-5, and benzene substituted with 1-3 hydroxy groups. In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is -CH=CH-(CH2) a -C(O)OH (wherein a is 0) and benzene substituted with 1 to 3 hydroxy groups.

[0217] In certain embodiments, the compound derived from, constituting, or purified from the chebula extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, or a metabolite of any of the foregoing.

[0218] In certain embodiments, the compound derived from, constituting, or purified from a chebula extract is chebulinic acid.

[0219] In certain embodiments, the active agent in the composition is chebulinic acid or a compound having at least a 95% similarity score with chebulinic acid (see, e.g., Table 2 above). The active agent may be or include an ellagitannin or a compound having at least a 95% similarity score with ellagitannin (see, e.g., Table 1 above).

[0220] According to certain embodiments, the composition may include a chebula extract enriched with one or more compounds that are components of the chebula extract. The compound components of the chebula extract, or the combination of compound components of the chebula extract, may be purified, for example, at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

[0221] In certain embodiments, the effective amount of a compound derived from, constituting, or purified from a chebula extract, and optionally an effective amount of an active agent of the composition, is between 1 and 100 mg, e.g., 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg of a compound derived from, constituting, or purified from a chebula extract.

[0222] The method and any of the above aspects may include administering a composition comprising two or more of an effective amount of an emblica extract or a compound component of an emblica extract, an effective amount of a fucus extract or a compound component of a fucus extract, and an effective amount of a chebula extract or a compound component of a chebula extract. According to certain embodiments, two or more of an emblica extract or a compound component of an emblica extract, a fucus extract or a compound component of a fucus extract, and a chebula extract or a compound component of a chebula extract may provide a synergistic effect in the treatment of a disease or condition.

[0223] The compositions disclosed herein may include or be enriched with one or more of emblica extract or compound components of emblica extract, fucus extract or compound components of fucus extract, chebula extract or compound components of chebula extract, BAMLET 10, BAMLET 50, ferulic acid, quercetin, urolithin A, pterostilbene, acadesin, embelin, EGCG, eriocitrin, gallic acid, gomsin A, lutein, luteolin, NAD, rutin, zeaxanthin, and melatonin.

[0224] The compositions disclosed herein may be administered in combination with and / or encapsulated in biomaterials. Biomaterials offer certain advantages for drug delivery, including favorable cell interaction promotion, relatively high drug loading content, controllable drug release, excellent passive and active targeting, biocompatibility, and low toxicity. Biomaterials can be classified into three basic categories: 1) natural biomaterials (e.g., extracellular vesicles, collagen, hyaluronic acid, fibrin), 2) synthetic biomaterials (e.g., polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), polyethylene glycol (PEG)), and 3) composite biomaterials (e.g., protein-polysaccharide composite biomaterials, nanocomposite biomaterials, sponges).

[0225] Biomaterials can be used to encapsulate and deliver the compounds disclosed herein, and optionally combination therapies, for the treatment of ovarian diseases and conditions. According to certain embodiments, the biomaterials described herein can be non-toxic, biocompatible, biodegradable, bioabsorbable, and / or capable of supporting cell and tissue regeneration without resulting inflammatory responses. Biomaterials may be used in conjunction with artificial ovarian constructs, follicular development, biomaterial encapsulation of cells and / or compositions, and delivery of natural extracellular vesicles. Exemplary biomaterials include extracellular vesicles, collagen, hyaluronic acid, synthetic biomaterials, fibrin, and alginate.

[0226] Extracellular vesicles (EVs) are a heterogeneous group of cell-derived membrane structures composed of exosomes (~50-150 nm) and microvesicles (~100-1000 nm), which carry bioactive materials and proteins in different body fluids and deliver their contents to recipient cells. Due to multiple advantages, including good biocompatibility with the immune system and low toxicity, EVs can be used as effective nanocarriers of the compositions disclosed herein for advanced drug delivery. Stem cell-derived EVs can be used to implant functional miRNAs and proteins into ovarian tissue. In addition, EVs can be easily isolated from stem cells of various origins and can carry biologically active molecules that can migrate to target ovarian cells to exhibit their therapeutic effects.

[0227] Collagen is the most abundant extracellular matrix protein in the animal kingdom, which transfers load in tissue and provides a highly biocompatible environment for cells. Collagen may be used as a scaffold for stem cell-based ovarian therapy. Such scaffolds may be used in conjunction with treatment for primary ovarian failure (POF) to increase the long-term retention of adipose-derived stem cells (ADSCs), and contribute to the restoration of ovarian function, including regular menstrual cycles, increased E2 levels, improved fertility, and successful clinical pregnancy. Collagen may be used to improve the targeted delivery of the compositions disclosed herein to ovarian tissue.

[0228] Hyaluronic acid (HA) is present in all vertebrates, is an important component of ECM in most mature tissues, and plays an important role in establishing the microenvironment that contributes to the development of follicles in the ovary. HA has excellent physicochemical properties and can be used for drug delivery. Self-crosslinked HA is a good cell scaffold for improving the transplantation of hepatocytes in the treatment of ovarian therapy. In addition, HA can be used to improve the targeted delivery of the compositions disclosed herein to ovarian tissue.

[0229] Fibrin (FIB) is a natural scaffold formed after tissue injury, which has excellent biocompatibility, controllability, biodegradability, and the ability to migrate cells and biomaterials to target sites. Primate oocytes derived from primary follicles cultured in fibrin-alginate 3D capsules can be used to restart meiosis for fertilization. Fibrin scaffolds can be used for follicle transplantation by controlling the release of growth factors and creating a continuous pathway for cell infiltration. Fibrin may be used to improve the targeted delivery of the compositions disclosed herein to ovarian tissue.

[0230] Synthetic biomaterials can be designed, selected, and / or tailored according to the physicochemical and mechanical properties of target biological tissue.PEG is a commonly used biocompatible polymer, and PEG hydrogels have been shown to provide a good microenvironment for ovarian follicles.Synthetic biomaterials can be designed to deliver the compositions disclosed herein to target ovarian tissue.

[0231] Alginate is a group of unbranched polysaccharides that is non-toxic, nutrient-rich, biocompatible and biodegradable in humans.Alginate can form soft hydrogels under physiological conditions, with pores large enough to allow nutrients and growth factors to pass freely, while trapping cells in the polymer network.Alginate can be used to improve the targeted delivery of the compositions disclosed herein to ovarian tissue.

[0232] The methods, and any of the above embodiments, may further include one or more of the steps of: (i) identifying a subject in need of treatment or administration; and (ii) providing a compound of the present disclosure or a pharmaceutical composition comprising a compound of the present disclosure.

[0233] In any of the above embodiments, and in any of the above aspects, when the term "preventing" is used, it can refer to at least partial inhibition, e.g., 10% inhibition, 20% inhibition, 30% inhibition, 40% inhibition, 50% inhibition, 60% inhibition, 70% inhibition, 80% inhibition, 90% inhibition, 95% inhibition, or greater than 95% inhibition.

[0234] In any of the above embodiments and in any of the above aspects, when the term "improving" is used, it can refer to at least a partial improvement, e.g., a 10% improvement, a 20% improvement, a 30% improvement, a 40% improvement, a 50% improvement, a 60% improvement, a 70% improvement, an 80% improvement, a 90% improvement, a 95% improvement, or an improvement of greater than 95%.

[0235] In any of the above embodiments, and in any of the above aspects, when the term "enhancing" is used, it can refer to at least a partial enhancement, e.g., a 10% enhancement, a 20% enhancement, a 30% enhancement, a 40% enhancement, a 50% enhancement, a 60% enhancement, a 70% enhancement, an 80% enhancement, a 90% enhancement, a 95% enhancement, or an enhancement of greater than 95%.

[0236] In any of the above embodiments, and in any of the above aspects, when the term "increasing" is used, it can refer to at least a partial increase, such as a 10% increase, a 20% increase, a 30% increase, a 40% increase, a 50% increase, a 60% increase, a 70% increase, an 80% increase, a 90% increase, a 95% increase, or an increase of more than 95%. It can refer to a 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 100-fold, or higher increase.

[0237] In any of the above embodiments and in any of the above aspects, the compound, or a pharma- ceutically acceptable form thereof, may be administered alone or as part of a pharmaceutical composition. A pharmaceutical composition may be formulated by combining a solution of the compound with a pharma- ceutically suitable carrier. The solution of the compound may contain 1-1,000 mg of the compound, for example, 10-600 mg, 20-500 mg, 30-200 mg, 50-100 mg, or 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1,000 mg of the compound.

[0238] The compound and a pharma- ceutical suitable carrier may be combined in a ratio of 1:5 to 5:1.The pharmaceutical composition may be formulated to have a concentration of the compound of 1 to 1,000 mg / ml, for example, 10 to 600 mg / ml, 20 to 500 mg / ml, 30 to 200 mg / ml, 50 to 100 mg / ml, or 1 mg / ml, 5 mg / ml, 10 mg / ml, 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 200 mg / ml, 300 mg / ml, 400 mg / ml, 500 mg / ml, 600 mg / ml, 700 mg / ml, 800 mg / ml, 900 mg / ml, or 1,000 mg / ml.

[0239] The pharmaceutical composition may be formulated to have a concentration of the compound of 0.01% to 2%, for example, 0.01%, 0.02%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, or 2%.

[0240] The pharmaceutical composition may be formulated to have a concentration of the compound of 2.5-50 μM, e.g., 2.5 μM, 5 μM, 10 μM, 25 μM, or 50 μM. The pharmaceutical composition may be formulated to have a concentration of the compound of 50-500 μM, e.g., 50 μM, 100 μM, 200 μM, 300 μM, 400 μM, or 500 μM.

[0241] The pharmaceutical composition may be formulated to have a concentration of the compound of 2.5-50 μM, e.g., 2.5 μg / L, 5 μg / L, 10 μg / L, 25 μg / L, or 50 μg / L. The pharmaceutical composition may be formulated to have a concentration of the compound of 50-500 μg / L, e.g., 50 μg / L, 100 μg / L, 200 μg / L, 300 μg / L, 400 μg / L, or 500 μg / L.

[0242] In any of the above embodiments and in any of the above aspects, the compound, or a pharma- ceutically acceptable form thereof, may be administered alone or as part of a cosmetic composition. The cosmetic composition may be formulated by combining a solution of the compound with a cosmetically suitable carrier. The solution of the compound may contain 1-1,000 mg of the compound, for example, 10-600 mg, 20-500 mg, 30-200 mg, 50-100 mg, or 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1,000 mg of the compound.

[0243] The compound and cosmetically suitable carrier may be combined in a ratio of 1:5 to 5:1. The cosmetic composition may be formulated to have a concentration of the compound of 1 to 1,000 mg / ml, for example, 10 to 600 mg / ml, 20 to 500 mg / ml, 30 to 200 mg / ml, 50 to 100 mg / ml, or 1 mg / ml, 5 mg / ml, 10 mg / ml, 20 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 200 mg / ml, 300 mg / ml, 400 mg / ml, 500 mg / ml, 600 mg / ml, 700 mg / ml, 800 mg / ml, 900 mg / ml, or 1,000 mg / ml.

[0244] In any of the methods and in any of the above embodiments, the compounds described herein are in the form of pharmaceutically acceptable salts, solvates or hydrates.Such compounds may be formulated as pharmaceutically acceptable salts, such as acid addition salts, and complexes thereof.The preparation of such salts may facilitate pharmacological use by modifying the physical characteristics of the drug, without hindering its physiological effect.Examples of useful modifications of physical properties include, but are not limited to, increasing solubility, making it easier to administer higher concentrations of the compound.

[0245] In any of the methods and in any of the above embodiments, the compounds or extracts described herein are administered topically, intravenously, intraperitoneally, parenterally, intramuscularly, orally, or via the airways. In any of the methods and in any of the above embodiments, the compounds or extracts are administered topically. In any of the methods and in any of the above embodiments, the compounds or extracts are administered parenterally, for example, by injection.

[0246] The compounds or extracts described herein may be administered locally, for example at a local site of a disease or condition. The compounds or extracts described herein may be formulated for local administration, for example to a target site of a disease or condition. The compounds or extracts described herein may be administered systemically. Systemic administration may be, for example, topically, intravenously, intraperitoneally, parenterally, intramuscularly, orally, or via the airways. The compounds or extracts described herein may be formulated for systemic administration.

[0247] In any of the methods, and in any of the above embodiments, the subject is a vertebrate. In any of the methods, and in any of the above embodiments, the subject is a mammal. In any of the methods, and in any of the above embodiments, the subject is a human. In any of the methods, and in any of the above embodiments, the subject is a non-human animal, e.g., a rodent, e.g., a mouse.

[0248] In some embodiments, the subject may be female. In some embodiments, the subject may be male. The subject may be characterized as one of the following ethnicities / races: Asian, Black or African American, Hispanic or Latino, Caucasian, or multiracial. The subject may be under 1 year old, or 1-5 years old, 5-10 years old, 10-20 years old, 20-30 years old, 30-40 years old, 40-50 years old, 50-60 years old, or over 60 years old. The subject may suffer from or be diagnosed with a condition that causes an increased risk of severe illness resulting from viral infection or where standard of care treatment causes an increased risk of severe illness resulting from viral infection. Such conditions include, for example, cancer, chronic kidney disease, chronic lung disease (such as chronic obstructive pulmonary disease (COPD), asthma, interstitial lung disease, cystic fibrosis, and pulmonary hypertension), dementia or other neurological conditions, type 1 or type 2 diabetes, Down's syndrome, cardiac conditions (such as heart failure, coronary artery disease, cardiomyopathy, or hypertension), immunosuppressive conditions, liver disease, overweight (e.g., having a body mass index of 25 or greater), obesity (e.g., having a body mass index of 30 or greater), pregnancy, sickle cell disease or thalassemia, current or former smoker, having undergone a solid organ or blood stem cell transplant, stroke or cerebrovascular disease, substance use disorder, concurrent viral infection, such as HIV infection, SARS-CoV-2 infection, influenza infection, or MERS infection, or any indication identified by the U.S. Centers for Disease Control and Prevention (CDC) as increasing the risk of severe illness or where standard treatment causes an increased risk of severe illness due to viral infection.

[0249] In any of the methods, and in any of the above embodiments, the compound or extract is administered in an effective amount. Suitable effective amounts are described in more detail herein. In any of the methods, and in any of the above embodiments, the compound or extract is administered in a therapeutically effective amount. In any of the methods, and in any of the above embodiments, the administering step may include administering a single dose of the compound or extract according to a course of treatment (where the dose may contain an effective amount). In any of the methods, and in any of the above embodiments, the administering step may include administering two or more doses of the compound or extract according to a course of treatment (where one or more doses may contain an effective amount). The amount of the compound or extract in each dose administered during a course of treatment need not be the same. For example, the administering step may include administering at least one loading dose and at least one maintenance dose during a course of treatment. Administration is described in more detail herein.

[0250] The compounds may be administered as a prophylactic treatment.The compounds may be administered as a cosmetic or therapeutic treatment.

[0251] The compounds disclosed herein can be used in a variety of applications, for example, cosmetic and / or therapeutic applications. The compounds may be administered in an effective amount for the intended use, for example, cosmetic or therapeutic applications. In some embodiments, the composition may contain a concentration or amount, for example, an effective amount, of the compound sufficient to have a desired cosmetic effect. In some embodiments, the composition may contain a concentration or amount, for example, an effective amount, of the compound sufficient to have a desired therapeutic effect.

[0252] The amount and / or frequency of administration may be sufficient to induce mitochondrial biogenesis. The amount and / or frequency of administration may be sufficient to treat, inhibit or prevent the progression of at least one of mitochondrial dysfunction, a disease or condition associated with mitochondrial dysfunction, or a symptom thereof, an age-related chronic condition associated with mitochondrial dysfunction, or a symptom thereof, and / or a decrease in energy level or vitality. The amount and / or frequency of administration may be sufficient to modify the score of a parameter on a qualitative scale, as scored by a subject or a clinical grader. The qualitative scale may include the following categories: none (best possible condition), mild, moderate, severe (worst possible condition). The qualitative scale may refer to perceived or actual energy level and / or vitality.

[0253] In some aspects, administering an effective amount of the compound may limit or inhibit the progression of at least one of mitochondrial dysfunction, a disease or condition associated with mitochondrial dysfunction, or a symptom thereof, an age-related chronic condition associated with mitochondrial dysfunction, or a symptom thereof, and / or a decrease in energy level or vitality.For example, an effective amount of the compound may slow down the progression of at least one of mitochondrial dysfunction, a disease or condition associated with mitochondrial dysfunction, or a symptom thereof, an age-related chronic condition associated with mitochondrial dysfunction, or a symptom thereof, and / or a decrease in energy level or vitality.In some embodiments, administering an effective amount of the compound may promote an increase in mitochondrial biogenesis, mitochondrial function, and / or energy level or vitality.

[0254] In some embodiments, administering an effective amount of the compound may increase the expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COX II. Administering an effective amount may decrease or inhibit the increase in the expression of at least one protein selected from FGF-21 and IL-6, or any cytokine associated with viral infection. Administering an effective amount may increase or inhibit the decrease in at least one of ATP-related respiration, maximum respiration, and spare capacity in a subject infected with SARS-CoV-2. Administering an effective amount may modulate, e.g., increase or decrease, the interaction of viral proteins with one or more host mitochondrial genes, such as MRPS2, MRPS5, MRPS25, MRPS27, NDUFAF1, NDUFB9, NDUFAF2, ATP1B1, ATP6V1A, ACADM, AASS, PMPCB, PITRM1, COQ8B, PMPCA, and Tomm70. Administration of an effective amount can increase the expression of ACE2.

[0255] The amount of the compound or composition can be effective for treating or preventing ovarian disease or condition in a subject.The amount of the composition can be effective for improving or enhancing fertility and / or extending reproductive life span.In certain embodiments, administering an effective amount, for example administering a therapeutically effective amount or a cosmetically effective amount, can induce mitochondrial biogenesis and / or improve mitochondrial function.For example, in some embodiments, administering an effective amount can increase mitochondrial mass of oocytes or prevent the decrease of mitochondrial mass of oocytes.

[0256] In some aspects, the compound may be administered prior to the onset of a disease or condition in a subject. The compound may be administered during the development of a disease or condition in a subject. The compound may be administered after at least partial reduction of a disease or condition in a subject.

[0257] The compounds may be administered in response to triggers or precursors of an ovarian disease or condition, such as prolonged or irregular menstrual cycles, reduced ovulatory events, and / or menstrual cramps or pain.

[0258] The compounds may be administered in response to triggers or warning signs of mitochondrial dysfunction or age-related conditions, such as aging, premature aging, habitual sleep conditions, weight loss, ultraviolet (UV) light exposure, treatment with chemical or therapeutic agents, such as chemotherapy and / or radiation therapy, smoking, dehydration, or immersion. A subject may be predisposed to a condition of mitochondrial dysfunction, for example, based on age, ethnicity, skin type, eye color, habits, or genetics.

[0259] The method may further include determining whether the subject is in need of treatment or administration.

[0260] The composition comprising the compound may additionally comprise a moisturizer, a deodorant, a fragrance, a colorant, an insect repellent, a cleaning agent, or a UV blocking agent. The composition may comprise a microsphere or a microcapsule.

[0261] The composition may be formulated for immediate release or sustained release.The composition may be formulated for controlled release or sustained release.For example, the composition may be formulated for sustained release over 6 hours, 12 hours, 18 hours, 24 hours, 36 hours, 48 ​​hours or longer.

[0262] The subject may be characterized as having normal mitochondrial function. The subject may be characterized as having reduced mitochondrial function. The subject may be characterized as having increased levels of circulating mtDNA, e.g., plasma mtDNA and / or cytoplasmic mtDNA.

[0263] The subject may be characterized by age-related reduced fertility, infertility, and / or ovarian aging. For example, in some embodiments, the subject or recipient subject is 40 years of age or older.

[0264] The subject may be characterized as experiencing premature aging or symptoms of premature aging.For example, the subject may be characterized as experiencing premature reproductive aging, which may include one or more of primary ovarian insufficiency (POI), premature extended or irregular menstrual cycle, premature reduction of ovulation events, premature menopause or perimenopause, and / or ovarian inflammation associated with premature reproductive aging.In such an embodiment, the subject may be under 40 years old.

[0265] In some embodiments, the subject may have a decreased ovarian reserve (DOR). In other embodiments, the subject may have a normal ovarian reserve (NOR).

[0266] The subject may be pre-menopausal, peri-menopausal, or post-menopausal.

[0267] The subject may suffer from reduced production of reproductive hormones, e.g., estrogen and / or progesterone, increased production of follicle-stimulating hormone (FSH), prolonged or irregular menstrual cycles, uterine or vaginal atrophy, e.g., loss of endometrial glands, scattered gonadal rupture of the uterus (apoptotic bodies), acute inflammation of the uterus, and / or reduced thickness of the vaginal epithelium, reduced ovulatory events, ovarian inflammation, and / or infertility.

[0268] The subject may be characterized by dysfunctional follicle formation, for example, follicle depletion or follicle dysfunction.For example, the subject may have a reduced number of tertiary follicles, for example, early antral, antral and / or preovulatory follicles, and / or a reduced number of corpus luteum (CL) follicles.

[0269] The subjects may be characterized by downregulation of estrogen receptor (ER) alpha and beta in the ovaries, reduced ESR1 or ESR2 gene expression, increased miR-206-3p RNA expression, reduced ovarian production of 17β-estradiol (E2), and / or an increase in lipid-laden ovarian stromal cells.

[0270] The subject may be characterized by reduced anti-Mullerian hormone (AMH) levels, e.g., less than about 80 ng / mL, increased follicle-stimulating hormone (FHL) levels, e.g., greater than about 10 IU / L, increased estradiol levels, and / or reduced antral follicle count (AFC), e.g., less than about 5-7 total follicles.

[0271] In certain embodiments, the subject may be characterized as suffering from one or more of obesity, diabetes, chronic inflammation, hyperglycemia, autoimmune disease, and / or poor lifestyle factors, such as smoking, alcohol use, drug use, exposure to chemicals (e.g., bisphenol A, advanced glycation end products (AGEs)) and / or radiation, nutritional deficiencies, such as increased intake of burnt foods, increased intake of saturated fats, and / or reduced intake of fruits and vegetables, and / or reduced exercise.

[0272] In certain embodiments, the subject may have undergone in vitro fertilization (IVF). In certain embodiments, the subject may have previously attempted IVF, either successfully or unsuccessfully. The subject may be a poor responder to IVF. In other embodiments, the subject may be a normal responder to IVF.

[0273] The subject may have mitochondrial DNA mutation in the oocyte that is fertilized to form embryo or recipient oocyte.The mutation may be somatic or genetic.In certain embodiments, the mutation may be, for example, T414G crossover mutation.

[0274] The subject may be characterized by a reduced level of mitochondrial DNA (mtDNA) in oocytes that are fertilized to form embryos. The subject may be characterized by reduced oocyte quality, e.g., chromosome misalignment and / or spindle abnormalities.

[0275] The subject may be characterized by an abnormality in the mitofusin (Mfn1) gene, an abnormality in the dynamic associated protein 1 (Drp1) gene, an abnormality in the caseinolytic peptidase P (Clpp) gene, an abnormality in the growth differentiation factor 9 (GDF9) gene, an abnormality in the fragile X mental retardation 1 (FMR1) gene, an abnormality in the transcription factor A (TFAM) gene, an abnormality in the mitochondrial DNA polymerase gamma (PolgA) gene, an abnormality in the mitochondrial inner membrane peptidase (IMP) gene, and / or an abnormality in the mitochondrial ABC transporter (MDR-1) gene. The subject may be characterized by decreased gene expression of PPARy, PGC-1a, PGC-1b, ERR, NRF-1, NRF-2, SIRT1, SIRT3, SIRT4, and / or SIRT5. The subject may be characterized by decreased AMP-activated protein kinase (AMPK) and / or PTEN-induced kinase 1 (PINK1) protein expression, and / or increased mammalian target of rapamycin (mTOR) protein expression.

[0276] The subject may be characterized by decreased gene expression of superoxide dismutase (SOD) and / or increased levels of Aldh3A2 enzyme. The subject may be characterized by decreased mRNA expression of Prdx3, Prdx4, and / or Txn2 enzymes.

[0277] In some embodiments, the subject may be characterized by neurological conditions.In some embodiments, the subject may suffer from Turner syndrome, galactosemia, and / or fragile X syndrome.In some embodiments, the subject may be characterized by progressive external ophthalmoplegia (POE), spinocerebellar ataxia with epilepsy (SCAE), and / or mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS).

[0278] In certain embodiments, the subject may be characterized by one or more of the following: inflammatory phenotype in skin, changes in mitochondrial protein expression, reduced expression of mitochondrial oxidative phosphorylation complex, reduced stability of mitochondrial oxidative phosphorylation complex, reduced collagen content of skin, increased epidermal thickness, increased epidermal hyperplasia, acanthosis, hyperkeratosis, increased expression of at least one gene selected from the group consisting of NF-κB, COX-2, INF-β1, CCL5, MMP1, MMP2, MMP9, MMP13, IGF1R, VEGF, and MRPS5, decreased expression of at least one gene selected from the group consisting of TIMP1, KLOTHO, COL1A1, MTCO2, TFAM, and VDAC, and increased inflammatory infiltrate in skin. The subject may be characterized as having a decreased expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COXII.

[0279] In some embodiments, the method may further comprise administering a second amount of the compound to the subject. The second amount may be administered as a second dose of the same formulation. The second amount may be administered in a different formulation. The second amount may be administered by the same route of administration in a different formulation, for example, a topical liquid or oil is administered together with a shampoo, conditioner, spray, cream, gel, body wash, soap, or lotion. The second amount may be administered by a different route of administration, for example, each amount may be administered independently topically, parenterally, or enterally.

[0280] In some embodiments, the method may further include administering a second compound to the subject. The second compound may be a compound that is a component of the same extract. The second compound may be a compound that is a component of a different extract. The second compound may be a compound that has a similarity score of at least 95% with the first compound. The second compound may be administered in the same formulation. The second compound may be administered in a different formulation.

[0281] The compound may be administered as part of a combination therapy. The method may include administering a second treatment in combination with the compound. The compound may be administered for a period before starting the second treatment. The compound may be administered in conjunction with the second treatment. The compound may be administered for a period after stopping the second treatment. The second treatment may be administered via an alternative mode of administration. The second treatment may be a cosmetic and / or therapeutic treatment.

[0282] The compound may be administered in combination with a second agent, e.g., a cosmetic or therapeutic agent, approved to treat or normally used to treat the disease or condition, or a symptom thereof.

[0283] The compound may be administered in combination with standard treatments for improving fertility.For example, the compound may be administered in combination with one or more of clomiphene, tamoxifen, letrozole, metformin, gonadotropin, gonadotropin releasing hormone, dopamine agonist, and other hormone treatments.

[0284] Compound may be administered in combination with standard treatment for treating ovarian disease or condition.For example, compound may be administered in combination with one or more of hormone therapy, such as estrogen, progesterone, testosterone, or its synthetic form, selective serotonin reuptake inhibitor (SSRI), gabapentin, clonidine, hormonal contraception, gonadotropin releasing hormone (GnRH) agonist or antagonist, heat therapy, pain relief, nonsteroidal anti-inflammatory drug (NSAID), such as ibuprofen or other analgesic, spironolactone, eflornithine, electrolysis, chemotherapy, radiation therapy, surgery, such as hysterectomy, bilateral salpingo-oophorectomy, and weight loss surgery, and lifestyle changes, such as weight loss, improved nutrition, and increased physical activity.

[0285] The compound may be administered in combination with caffeine, B vitamins (e.g., B1, B2, B3, B5, B6, B8, B9 and / or B12), vitamin C, vitamin D, iron, magnesium, and / or zinc.

[0286] The compounds may be administered in combination with UV blockers, moisturizers, sunscreens, wrinkle creams, retinoids, alpha-hydroxy acids, beta-hydroxy acids, squalene, antioxidants, tretinoin, glycosaminoglycans (GAGs), lactic acid, malic acid, citric acid, tartaric acid, hydroquinone, kojic acid, L-ascorbic acid, licorice extract, N-acetylglucosamine, niacinamide, soy, dermal fillers or injectables, such as hyaluronic acid or calcium hydroxylapatite, botulinum toxin, laser resurfacing procedures, ultrasound therapy, chemical peels, such as glycolic acid peels, trichloroacetic acid or salicylic acid, or skin peeling procedures.

[0287] The compounds may also be administered in conjunction with a surgical procedure, for example, fallopian tube surgery, laparoscopic surgery to remove or destroy cysts or submucosal fibroids, or laparoscopic ovarian drilling.

[0288] The compound may be administered in combination with an antioxidant. Exemplary antioxidants include CoQ10, vitamin C, vitamin E, carotenoids, such as beta-carotene, minerals, such as selenium and manganese, glutathione, lipoic acid, flavonoids, beta flavinoids, phenols, polyphenols, phytoestrogens, mitoquinol mesylate, and ubiquinone.

[0289] The fucus compound may be administered in combination with an emblica extract or a compound component of an emblica extract. The fucus compound may be administered in combination with a chebula extract or a compound component of a chebula extract. The emblica compound may be administered together with a fucus extract or a compound component of a fucus extract. The emblica compound may be administered together with a chebula extract or a compound component of a chebula extract. According to certain embodiments, the combination of two or more of the emblica compound, the fucus compound, and the chebula compound may provide a synergistic effect in the treatment of a disease or condition.

[0290] According to one or more embodiments, an effective amount of the compound may be administered to the face of a subject. According to one or more embodiments, the compound may be administered to the scalp of a subject. According to one or more embodiments, the compound may be administered to the body of a subject. For example, the compound may be applied to one or more of the forehead, eye area, neck, scalp, head, shoulders, arms, hands, legs, underarms, torso, chest, feet, knees, ankles, back, buttocks, or genitalia of a subject.

[0291] In accordance with one or more embodiments, an effective amount of the compound may be administered enterally or parenterally.

[0292] In vitro treatment methods, such as in vitro fertilization (IVF) According to certain embodiments, the subject may have undergone in vitro fertilization (IVF). The subject may have previously attempted IVF, with or without success. The subject may be a poor responder to IVF. In other embodiments, the subject may be a normal responder to IVF.

[0293] During IVF, oocytes are typically retrieved from the ovaries of a subject and placed in vitro with active motile sperm to generate one or more embryos. Viable embryos can be selected for implantation into the uterus of the subject or a surrogate mother. Additional viable embryos can be stored for future implantation. The methods disclosed herein can include improving embryo development or improving embryo fertility.

[0294] Thus, in some embodiments, the embryos may be produced by IVF. The method may include measuring the mitochondrial content of the embryos and selecting embryos that respond to the measured mitochondrial content within a predetermined range. The predetermined range may correspond to a healthy range, for example, a range associated with normal ovarian reserve (NOR) and fertility.

[0295] The method may include administering the composition disclosed herein to a subject or embryo.For example, the method may include treating an embryo by introducing the composition into the culture medium of the embryo during development.The composition may be introduced in an amount effective to improve embryo development or fertility.The method may include introducing an antioxidant into the culture medium in combination with the composition.

[0296] In certain embodiments, the method may include transferring ooplasm from a donor oocyte to a recipient oocyte of a subject to be fertilized to form an embryo. In general, the donor oocyte may have a higher mitochondrial DNA (mtDNA) content than the recipient oocyte of the subject. The donor oocyte may be autologous. The donor oocyte may be allogeneic. The donor oocyte may be xenogeneic. In such embodiments, the composition may be administered to the donor subject, the recipient subject, and / or the embryo.

[0297] The subject may have a mitochondrial DNA mutation in the oocyte that is fertilized to form an embryo. The mutation may be somatic or genetic. In certain embodiments, the mutation may be, for example, a T414G crossover mutation. In some embodiments, the subject may be characterized by a reduced level of mitochondrial DNA (mtDNA) in the oocyte that is fertilized to form an embryo. The subject may be characterized by a reduced oocyte quality, for example, chromosome misalignment and / or spindle abnormalities.

[0298] In some embodiments, the ooplasmic transfer may be a complete transfer. In other embodiments, the ooplasmic transfer may be partial. Partial ooplasmic transfer may be by electrofusion or direct ooplasmic injection.

[0299] Eoplasmic transfer may be performed by one of several methods. In some embodiments, ooplasmic transfer may be performed by modified intracytoplasmic sperm injection (ICSI). In some embodiments, ooplasmic transfer may be performed by autonomous germline mitochondrial energy transfer (AUGMENT) to oocytes. Eoplasmic transfer may be performed by nuclear genome transfer, for example, oocyte spindle transfer, germinal vesicle (GV) transfer, pronuclear transfer (PNT), or polar body nuclear transfer (PBNT).

[0300] In some embodiments, the method may include using CRISPR / Cas 9 gene editing technology on the mitochondrial DNA (mtDNA) of the oocyte of the subject to be fertilized to form an embryo. In embodiments involving ooplasmic transfer, the method may include using CRISPR / Cas 9 gene editing technology on the mtDNA of the donor oocyte or the recipient oocyte.

[0301] In some embodiments, the method may include introducing stem cell-generated mitochondrial DNA (mtDNA) into a subject's oocyte to be fertilized to form an embryo. Thus, in certain embodiments, mtDNA transfer may occur without a donor oocyte.

[0302] Therefore, according to another embodiment, a preparation is provided that contains mtDNA as a therapeutic agent. The preparation may include an effective amount of donor mtDNA in a therapeutically acceptable carrier. The mtDNA may be edited by CRISPR / Cas 9. The mtDNA may be made by stem cells. The mtDNA may be in the form of a complete ooplasmic transfer or a partial ooplasmic transfer.

[0303] Also disclosed are kits that include preparations that include mtDNA. The kits may additionally include a compound of the present disclosure, or a pharma- ceutically acceptable form thereof.

[0304] Dosage and Administration According to the method, a compound of the disclosure may be administered to a subject (or contacted with a cell of a subject) in an effective amount.

[0305] In certain embodiments, a therapeutically effective amount of a compound of the present disclosure ranges from about 0.05 mg / kg / day to about 50 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 40 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 30 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 20 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 10 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 8 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 6 mg / kg / day. In certain embodiments, an effective amount ranges from about 0.05 mg / kg / day to about 4 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 3 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 2 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 1 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 0.8 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 0.6 mg / kg / day. In certain embodiments, the effective amount is in the range of about 0.05 mg / kg / day to about 0.4 mg / kg / day. In certain embodiments, the daily amounts described above are administered according to the course of treatment and may be administered in a single dose or two or more doses per day. The daily amounts described above may be administered according to a course of treatment, and may be administered in one dose (qd) or two doses (bid) on each day, where the amount of compound of the present disclosure in each dose need not be the same.

[0306] In certain embodiments, each dose is administered according to a course of treatment. As used herein, the term "dose" refers to the amount of the compound of the present disclosure administered at a given time according to a course of treatment. For example, if the course of treatment for the compound of the present disclosure is bid (2 times / administration per day) for 7 days, then two administrations on each of days 1-7 each include administering a dose of the compound of the present disclosure (2 doses on each day). In certain embodiments, the dose is administered qd (1 time / administration per day) according to the course of treatment. In certain embodiments, the dose is administered bid according to the course of treatment. In certain embodiments, the dose is administered tid (3 times / administration per day) according to the course of treatment.

[0307] When two or more doses are administered on a given day according to a course of treatment, each dose administered according to a course of treatment may contain the same amount of the compound of the present disclosure, or one or more doses administered according to a course of treatment may contain a greater or lesser amount of the compound of the present disclosure compared to another dose administered according to a course of treatment. For example, if a course of treatment for a compound of the present disclosure is 7 days bid, the first dose administered on day 1 may contain a first amount (i.e., 2 mg / kg) and the second dose administered on day 1 may contain a second amount (i.e., 0.5 mg / kg). As another example, if the course of treatment for a compound of the disclosure is 7 days bid, the first dose administered on day 1 may contain a first amount (i.e., 2 mg / kg), the second dose administered on day 1 may contain a second amount (i.e., 0.5 mg / kg), two doses administered on each of days 2-4 may contain a second amount, and two doses administered on each of days 5-7 may contain a third amount (i.e., 1 mg / kg).

[0308] A dose may be further divided into subdoses. Any given dose may be delivered in a single unit dosage form or in two or more unit dosage forms. For example, a dose given by IV administration may be provided as a single IV injection (i.e., a single 5 mg / kg IV injection) or as two or more IV injections administered in sequence (i.e., two 2.5 mg / kg IV injections). Further, a sub-dose may be, for example, a number of separate loosely spaced administrations, such as multiple inhalations from an inhaler, multiple drops applied to the eye, or multiple tablets for oral administration.

[0309] In certain embodiments, two or more doses of the disclosed compound are administered during the course of treatment. Thus, in the methods described herein, the methods may include administration of multiple doses during the course of treatment. In certain embodiments, the course of treatment may range from several months to several years. In certain embodiments, the course of treatment may range from 2 days to 1 month, 2 days to 3 weeks, 2 days to 2 weeks, or 2 days to 1 week. In certain embodiments, the course of treatment may range from 1 year to 20 years or longer. In certain embodiments, the dose is delivered at least once per day (i.e., 1 to 3 times) during the course of treatment. In certain embodiments, the dose is not administered daily during the course of treatment (e.g., the dose should be administered at least once per day (1 timer) every other day, every 3 days, weekly, or monthly during the course of treatment). Furthermore, the amount of the disclosed compound in each dose need not be the same, as discussed above. In certain embodiments, one or more, or all, of the above doses contain an effective amount of a compound of the disclosure.

[0310] In one embodiment, the course of treatment may include administering at least one dose as a loading dose and at least one dose as a maintenance dose, where the loading dose contains a greater amount of the disclosed compound compared to the maintenance dose (e.g., but not limited to, 2-10 times higher). In one aspect of this embodiment, the loading dose is administered first, either as a single dose or as two or more doses, followed by one or more maintenance doses throughout the remaining course of treatment. For example, for a course of treatment weekly for 10 years qd, a loading dose of 3 mg / kg may be administered as the first dose in week 1 of the course of treatment, followed by a maintenance dose of 0.5 mg / kg administered weekly for the remaining course of treatment. Furthermore, the loading dose may be given as a dose that is not the first dose administered during the course of treatment. For example, the loading dose may be administered as the first dose in a week and as a dose in one or more additional weeks (e.g., weeks 10 and 20).

[0311] In one embodiment, a course of treatment may include administering a first dose formulated in a first composition and administering at least one second or subsequent dose formulated in a second composition.The first composition and the second composition may be in the same formulation.The first composition and the second composition may be in different formulations.

[0312] Pharmaceutical, cosmetic and / or food compositions Provided is a pharmaceutical composition that contains an amount, for example an effective amount, of the compound of the present disclosure.In one embodiment, such a pharmaceutical composition contains a therapeutically effective amount of the compound of the present disclosure.In addition, other active agents may be included in such a pharmaceutical composition.The additional active agents included can be selected based on the disease or condition to be treated.

[0313] The disclosed pharmaceutical composition may comprise one or more of the disclosed compounds, alone or in combination with additional active agents, in combination with pharma- ceutically acceptable carriers and / or excipients, and / or in combination with cosmetically acceptable carriers and / or excipients.Such pharmaceutical compositions may be used in the manufacture of medicaments for use in the methods described herein.The disclosed compounds are useful in both free form and pharma- ceutically acceptable form, for example, pharma-ceutically acceptable salts.

[0314] Pharmaceutically acceptable carriers and / or excipients and / or cosmetically acceptable carriers and / or excipients are well known to those skilled in the art. The choice of carriers and / or excipients is determined in part by the specific compound(s) and by the specific method used to administer the composition of the compound. Thus, there are a wide variety of suitable formulations of the compounds of the present disclosure. The following methods and excipients are merely illustrative and in no way limiting. Suitable carriers and excipients include solvents such as water, alcohol, and polyethylene glycol, solid absorbents and diluents, surface active agents, suspending agents, tableting binders, lubricants, flavorings, and coloring agents. Pharmaceutically and / or cosmetically acceptable carriers may include polymers and polymer matrices. Examples of acceptable carriers include, among others, carboxymethylcellulose, crystalline cellulose, glycerin, gum arabic, lactose, magnesium stearate, methylcellulose, powder, saline, sodium alginate, sucrose, starch, talc, and water. Typically, acceptable carriers are chemically inert to the active agents in the composition and have no detrimental side effects or toxicity under the conditions of use.

[0315] Surfactants, such as detergents, are also suitable for use in the formulation. Specific examples of surfactants include polyvinylpyrrolidone, polyvinyl alcohol, copolymers of vinyl acetate and vinylpyrrolidone, polyethylene glycol, benzyl alcohol, polyoxyethylated esters of mannitol, glycerol, sorbitol, or sorbitan; lecithin or sodium carboxymethylcellulose; or acrylic derivatives, such as methacrylates; non-ionic surfactants, such as alkaline stearates, in particular sodium, potassium or ammonium stearate; calcium or triethanolamine stearate; alkyl sulfates, in particular sodium lauryl sulfate and sodium cetyl sulfate; sodium dodecylbenzenesulfonate or sodium dioctyl sulfosuccinate; or fatty acids, in particular those derived from coconut oil; cationic surfactants, such as those of the formula N+R'R''R''''R''''Y-, where the R groups are the same or different, as appropriate. and Y- is a hydroxylated hydrocarbon group and Y- is the anion of a strong acid, such as the anion of a halide, sulfate, and sulfonate; cetyltrimethylammonium bromide, which is one of the cationic surfactants which may be used, amine salts of the formula N+R'R''R''', where the R groups are the same or different, optionally, hydroxylated hydrocarbon groups; octadecylamine hydrochloride, which is one of the cationic surfactants which may be used, nonionic surfactants, such as esters of sorbitan, which are optionally polyoxyethylated, in particular polysorbate 80, or polyoxyethylated alkyl ethers; polyethylene glycol stearate, polyoxyethylated derivatives of castor oil, polyglycerol esters, polyoxyethylated fatty alcohols, polyoxyethylated fatty acids or copolymers of ethylene oxide and propylene oxide, amphoteric surfactants, such as substituted lauryl compounds of betaine.

[0316] Exemplary pharma- ceutically acceptable carriers and / or excipients and / or cosmetically acceptable carriers and / or excipients include water, silica, glycerin, dimethicone, butylene glycol, pentylene glycol, ethoxydiglycol, polyacrylate-13, pentapeptide-34 trifluoroacetate, polyisobutene, lysolecithin, sclerotium gum, pullulan, polysorbate 20, diethylhexyl syringylidenemalonate, caprylyl glycol, glyceryl stearate, PEG-100 stearate, cetearyl alcohol, butyrospermum parkii, glyceryl stearate ... Parkii (Shea) Butter, Acetyl Tetrapeptide-2, Betaine, Melanin, Tocopheryl Acetate, Tocopherol, Hydroxyacetophenone, Caprylic / Capric Triglyceride, Batyl Alcohol, C12-15 Alkyl Benzoate, Panthenol, Ceteareth-20, Xanthan Gum, Ethylhexylglycerin, Disodium EDTA, Propanediol, Caprylyl Glycol, Potassium Sorbate, Sorbic Acid, and Phenoxyethanol.

[0317] The compounds of the present disclosure, and pharmaceutical compositions containing such compounds as described in this disclosure, can be administered by any conventional method available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in combination with additional therapeutic agents.

[0318] In one embodiment, the compound of the present disclosure is administered in effective amount, whether alone or as part of pharmaceutical composition.The effective amount and dosage administered will of course vary according to known factors, such as the pharmacodynamic characteristics of specific drug and its mode and route of administration, recipient's age, health condition and weight; disease status or condition severity and stage; type of combined treatment; frequency of treatment; and desired effect.

[0319] The total amount of compound administered will also be determined by the route, timing and frequency of administration, as well as the existence, nature and extent of any adverse side effects that may accompany the administration of the compound and the desired physiological effect. It will be recognized by those skilled in the art that various conditions or disease states, particularly chronic conditions or disease states, may require long-term treatment, including multiple administrations.

[0320] In these pharmaceutical or cosmetic compositions, the compound(s) of the present disclosure are typically present in an amount of about 0.5-95% by weight based on the total weight of the composition, or about 0.1-99.9% by weight based on the total weight of the composition. Multiple dosage forms may be administered as part of a single treatment.

[0321] The active agent can be administered enterally in solid dosage form, for example capsules, tablets, and powders, or in liquid dosage form, for example emulsions, elixirs, syrups, and suspensions.It can also be administered parenterally in sterile liquid dosage form.The compound(s) of the present disclosure can also be administered intranasally (nose drops) or by inhalation via the pulmonary system, for example by aerosol-based metered dose inhaler or dry powder inhalation device.Other dosage forms include topical administration, for example transdermal administration via patch mechanism or ointment.

[0322] Formulations suitable for enteral or oral administration may be liquid solutions, such as an effective amount of compound(s) dissolved in a diluent, such as emulsion, water, saline, buffer solution, infant formula, other suitable carriers, or combinations thereof. Formulations suitable for enteral or oral administration of the compounds of the present disclosure are known in the art, as exemplified by Shaji, et al., Indian J Pharm Sci. 2008 May-Jun; 70(3): 269-277;Bruno, et al., Ther Deliv. 2013 Nov; 4(11): 1443-1467;Ibrahim, et al., DARU Journal of Pharmaceutical Sciences, 2020, 28, 403-416. The compound(s) can then be mixed with the diluent immediately prior to administration. In alternative embodiments, formulations suitable for enteral or oral administration may be capsules, sachets, tablets, lozenges, and troches. In each embodiment, the formulation may contain a predetermined amount of the compound(s) of the present disclosure as solids or granules, powders, suspensions, and suitable emulsions. Liquid formulations may contain diluents such as water, and alcohols, e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, and polyethylene alcohol, either with or without the addition of an acceptable surfactant, suspending agent, or emulsifying agent. Capsule forms may be of the usual hard- or soft-shelled gelatin type, containing, for example, surfactants, lubricants, and inert fillers, e.g., lactose, sucrose, calcium phosphate, and corn starch. Tablet forms may contain one or more of the following: lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, as well as other excipients, colorants, diluents, buffers, disintegrants, humectants, preservatives, flavorings, and pharmacologically compatible carriers.

[0323] Lozenge forms can contain the active ingredient in a flavoring, usually sucrose and acacia, or tragacanth, and can be pastilles containing the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia, emulsions, and gels, containing in addition to the active ingredient carriers, such as those known in the art.

[0324] Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the patient's blood, and aqueous and non-aqueous sterile suspensions which may contain suspending agents, solubilizing agents, thickening agents, stabilizers, and preservatives. The compound(s) may be administered in an acceptable carrier, with or without the addition of an acceptable surfactant, e.g., a soap or detergent, a suspending agent, e.g., pectin, carbomer, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants, in a physiologically acceptable diluent in a sterile liquid or mixture of liquids including water, saline, aqueous dextrose and related sugar solutions, alcohols, e.g., ethanol, isopropanol, or hexadecyl alcohol, glycols, e.g., propylene glycol or polyethylene glycols, e.g., poly(ethylene glycol) 400, glycerol, ketals, e.g., 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides.

[0325] The oil that can be used in parenteral formulation includes petroleum, animal oil, vegetable oil, or synthetic oil.Specific examples of oil include peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum, and mineral oil.Suitable fatty acid for use in parenteral formulation includes oleic acid, stearic acid, and isostearic acid.Ethyl oleate and isopropyl myristate are examples of suitable fatty acid ester. Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents, such as, for example, dimethyldialkylammonium halides and alkylpyridinium halides; (b) anionic detergents, such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; (c) nonionic detergents, such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene polypropylene copolymers; (d) amphoteric detergents, such as, for example, alkyl beta-aminopropionates, and 2-alkylimidazoline quaternary ammonium salts; and (e) mixtures thereof.

[0326] Parenteral formulations typically contain about 0.5% to about 50% by weight of the compound(s) in solution. Suitable preservatives and buffers can be used in such formulations. To minimize or eliminate irritation at the site of injection, such compositions may contain one or more non-ionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such formulations ranges from about 5% to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate, and polymeric adducts of ethylene oxide and hydrophobic bases formed by the condensation of propylene oxide with propylene glycol.

[0327] The compound(s) of the present disclosure can be formulated into aerosol preparations that are administered via nasal or pulmonary inhalation.These aerosol preparations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, and nitrogen.Such aerosol preparations can be administered by metered dose inhalers.They can also be formulated as pharmaceuticals for non-pressurized preparations, such as nebulizers or atomizers.

[0328] The compound(s) of the present disclosure, alone or in combination with other suitable components, may be administered in aqueous solution as nasal or pulmonary spray, or may be dispensed in spray form by various methods known to those skilled in the art. A system for dispensing liquid as nasal spray is disclosed in U.S. Pat. No. 4,511,069. The formulation may be present in a multi-dose container, such as the closed dispensing system disclosed in U.S. Pat. No. 4,511,069. Additional aerosol delivery forms may include, for example, compressed air nebulizers, jet nebulizers, ultrasonic nebulizers, and piezoelectric nebulizers, which deliver active agents dissolved or suspended in pharmaceutical solvents, such as water, ethanol, or mixtures thereof.

[0329] The nasal and pulmonary solutions of the present disclosure typically include a drug or drugs to be delivered and may be formulated with a surfactant, such as a non-ionic surfactant (e.g., polysorbate-80), and one or more buffers, as appropriate. In some embodiments, the nasal spray solution further includes a propellant. The pH of the nasal spray solution is about pH 3.0-6.0, or 4.5+ / -0.5, as appropriate. Suitable buffers for use within these compositions are as described above or otherwise known in the art. Other components, including preservatives, surfactants, dispersants, or gases, may be added to enhance or maintain chemical stability. Suitable preservatives include, but are not limited to, phenol, methylparaben, paraben, m-cresol, thiomersal, chlorobutanol, benzylalkonium chloride, and the like. Suitable surfactants include, but are not limited to, oleic acid, sorbitan trioleate, polysorbates, lecithin, phosphatidylcholine, and various long-chain diglycerides and phospholipids. Suitable dispersants include, but are not limited to, ethylenediaminetetraacetic acid, etc. Suitable gases include, but are not limited to, nitrogen, helium, chlorofluorocarbons (CFCs), hydrofluorocarbons (HFCs), carbon dioxide, air, etc.

[0330] In alternative embodiments, nasal and pulmonary formulations are administered as dry powder formulations, usually lyophilized, containing the dried active agent in a form of suitable particle size or within a suitable particle size range for intranasal delivery. The minimum particle size suitable for deposition in the nasal or pulmonary passages is often about 0.5 μm mass median aerodynamic diameter (MMEAD), usually about 1 μm MMEAD, more typically about 2 μm MMEAD. The maximum particle size suitable for deposition in the nasal passages is often about 10 μm MMEAD, usually about 8 μm MMEAD, more typically about 4 μm MMEAD. Intranasal and pulmonary respirable powders within these size ranges can be produced by a variety of conventional techniques, such as jet milling, spray drying, solvent precipitation, supercritical fluid condensation, etc. These dry powders of suitable MMEADs can be administered to a patient via a conventional dry powder inhaler (DPI) that relies on the patient's breathing during pulmonary or nasal inhalation to disperse the powder into an aerosolized amount. Alternatively, the dry powder may be administered via an air-assisted device, e.g., a piston pump, that uses an external powder source to disperse the powder into an aerosolized amount.

[0331] To formulate a composition for nasal or pulmonary delivery, the active agent can be combined with various pharma- ceutically and / or cosmetically acceptable additives, as well as bases or carriers for dispersing the active agent(s). Desired additives include, but are not limited to, pH control agents, such as arginine, sodium hydroxide, glycine, hydrochloric acid, citric acid, and the like. In addition, local anesthetics (e.g., benzyl alcohol), isotonicity agents (e.g., sodium chloride, mannitol, sorbitol), absorption inhibitors (e.g., Tween® 80), solubility enhancers (e.g., cyclodextrin and its derivatives), stabilizers (e.g., serum albumin), and reducing agents (e.g., glutathione). When the composition for nasal or pulmonary delivery is liquid, the tonicity of the formulation is typically adjusted to a value that does not induce substantially irreversible tissue damage in the nasal mucosa at the site of administration, as measured with reference to the tonicity of 0.9% (w / v) saline. Generally, the tonicity of the solution is adjusted to a value of about 1 / 3 to 3, more typically 1 / 2 to 2, and most often 3 / 4 to 1.7.

[0332] The compound(s) of the present disclosure may be dispersed in a base or vehicle that may contain a hydrophilic compound capable of dispersing the active agent and any desired additives. The base may be selected from a wide range of suitable carriers, including, but not limited to, polycarboxylic acids or their salts, copolymers of carboxylic anhydrides (e.g., maleic anhydride) with other monomers (e.g., methyl (meth)acrylate, acrylic acid, etc.), hydrophilic vinyl polymers such as polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone, cellulose derivatives such as hydroxymethylcellulose, hydroxypropylcellulose, etc., and natural polymers such as chitosan, collagen, sodium alginate, gelatin, hyaluronic acid, and non-toxic metal salts thereof. In many cases, biodegradable polymers are selected as the base or carrier, such as polylactic acid, poly(lactic acid-glycolic acid) copolymers, polyhydroxybutyric acid, poly(hydroxybutyric acid-glycolic acid) copolymers, and mixtures thereof. Alternatively or in addition, synthetic fatty acid esters, such as polyglycerin fatty acid esters, sucrose fatty acid esters, etc., may be used as carriers. Hydrophilic polymers and other carriers can be used alone or in combination, and enhanced structural integrity can be imparted to the carrier by partial crystallization, ionic bonding, crosslinking, etc. The carrier can be provided in various forms, including liquid or viscous solutions, gels, pastes, powders, microspheres and films for direct application to the nasal mucosa. The use of selected carriers in this context can result in enhanced absorption of active agents.

[0333] The compounds of the present disclosure may be formulated as described in International Patent Application No. PCT / US2022 / 020983, filed March 18, 2022, entitled "Compositions and Methods for Improving Mitochondrial Function," the entire disclosure of which is incorporated herein by reference in its entirety for all purposes.

[0334] The compounds of the present disclosure may be formulated in nanoparticle-based delivery carriers. The nanoparticle delivery systems disclosed herein may provide benefits in mitochondrial targeted delivery and improved therapeutic potential of the compounds. Nanoparticle formulations have been shown to efficiently transport drug molecules in their original form, solubilize hydrophobic drug molecules, enhance the half-life of the molecules, and reduce the side effects and immunogenicity caused by the molecules. The nanoparticle delivery carriers disclosed herein may be selected or designed to provide enhanced skin penetration, higher stability, site-specific targeting, e.g., efficient delivery of cargo inside the mitochondrial matrix, high entrapment efficiency, and / or time-controlled, e.g., delayed or sustained release of the compound. The properties of the nanoparticle delivery carriers that may be designed to efficiently deliver the compound of the formulation to the target site of the subject include surface chemistry, coating, structure, size, ability to aggregate, and solubility. Exemplary nanoparticles are described in "Nanotherapeutic Approaches to Target Mitochondria in Cancer," (Mani, 2021) and "Role of Nanotechnology in Cosmeceuticals: A Review of Recent Advances," (Kaul, 2018), each of which is incorporated by reference in its entirety for all purposes.

[0335] Nanoparticle delivery carriers may include and / or be functionalized with carbon-based nanomaterials, liposomal delivery vehicles, polymeric nanocarriers, micelles, dendrimers, lipophilic cations, solid-lipid nanoparticles (SLN), peptide-based nanomaterials, nanostructured lipid carriers (NLC), niosomes, nanoemulsions, metal nanoparticles, nanospheres, polymersomes, cubosomes, and combinations thereof. Delivery carriers may be designed for mitochondrial targeting or specific cell type targeting. Exemplary mitochondrial targeting agents include antibodies, polymeric functional moieties such as PEG, lipophilic cations such as triphenylphosphine (TPP), and peptides such as mitochondrial penetrating peptides (MPP). Delivery carriers may be formed from targeting moieties, e.g., encapsulating or conjugating with compounds disclosed herein, and / or delivery carriers may be functionalized with surface targeting moieties. The nanoparticle carriers may be sized to have an average size of about 10-5000 nm, e.g., 10-50 nm, 10-100 nm, 50-500 nm, 50-100 nm, 100-500 nm, 500-1000 nm, or 1000 nm-5000 nm, which may be selected based on the target tissue, the compound to be delivered, and other properties of the nanomaterial.

[0336] Exemplary carbon-based nanomaterials include carbon dots (C-dots or CDs), carbon nanotubes (CNTs), graphene derivatives, nanodiamonds (NDs), quantum dots (QDs), and magnetic nanoparticles (MNPs). Carbon nanoparticles may be formed into different shapes, including, for example, spherical, elliptical, tubular, angular, and combinations thereof.

[0337] CNTs are graphene sheets formed into cylindrical tubes. CNTs have demonstrated low toxicity profiles, good biocompatibility, and targeted accumulation. The CNTs of the present disclosure can be single-walled carbon nanotubes (SWCNTs) and / or multi-walled carbon nanotubes (MWCNTs). SWCNTs have smaller diameters ranging from 1-10 nm. MWCNTs have larger diameters ranging from 2-50 nm. CNTs may be functionalized with targeting sequences. For example, CNTs may be functionalized to target mitochondria and / or specific cell types. Graphene derivatives also include two-dimensional carbon isotopes. Graphene derivatives may be designed to exhibit specific physicochemical properties, such as high surface area and selected multifaceted surface properties. Graphene derivatives may be functionalized, e.g., surface functionalized, for targeted delivery to mitochondria and / or specific cell types. CNTs have been successfully used in hair colorants and cosmetic hair care formulations.

[0338] Nanodiamonds have been shown to provide high affinity, biocompatibility, and non- or low-cytotoxicity for biomolecules. Nanodiamonds, quantum dots, and magnetic nanoparticles may be conjugated with the compounds disclosed herein for targeted delivery. Certain MNPs may be designed to encapsulate the compounds disclosed herein. QDs are generally formed from a semiconducting core layered with a shell designed to provide selected physical and chemical characteristics. MNPs are formed from a magnetic core, e.g., an iron oxide material core, and a surface coating designed to improve stability and biocompatibility in physiological environments. As such, MNPs and QDs are highly adaptable for their selected uses.

[0339] Liposomal delivery vehicles are typically formed from encapsulated spherical vesicles composed of lipid bilayers with an internal bilayer and an internal aqueous core region. Liposomes may have a unilamellar or multilamellar structure. Liposomes may be designed to have effective surface chemistry for targeted delivery and / or controlled release delivery. For example, engineered liposomes have shown improved cellular update and accumulation of delivered compounds in mitochondria. Agents such as antioxidants, e.g., carotenoids, CoQ10, lycopene, and vitamins A, E, and K, may be incorporated into liposomes to amplify physical and chemical stability. Liposomes may be formulated with phosphatidylcholine to provide moisturizing properties to skin and hair care products. Vegetable phospholipids and soybean phospholipids may be used with topical formulations due to their high content of esterified essential fatty acids. For example, when applied with active agents, the barrier function of the skin is increased and water loss is reduced. Certain liposomes have been shown to have effects on reducing wrinkles, decreasing breakouts in the treatment of acne, and increasing skin smoothness.

[0340] Niosomes are vesicles with a bilayer structure composed of self-assembled hydrated non-ionic surfactants. Niosomes may have cholesterol incorporated in their lipids or may be cholesterol-free. Niosomes may be formulated as multi- or unilamellar structures, encapsulating the compounds disclosed herein by the membrane formed when the surfactant macromolecules are organized as bilayers. Exemplary non-ionic surfactants include span®, tween®, brij®, alkylamides, sorbitan esters, crown esters, polyoxyethylene alkyl ethers, and steroid-like surfactants. Niosomes may encapsulate the compounds disclosed herein to provide extended systemic circulation and enhanced penetration to target tissues. Niosomes in cosmetic and skin care applications provide skin penetration, increased stability of entrapped ingredients, and improved bioavailability of compounds that are typically poorly absorbed. Niosomes may be designed for targeted applications by controlling the surfactant nature and structure, membrane composition, and temperature of hydration, which affect the size and shape of the particle. Special niosomes called proniosomes may be used. Proniosomes are non-ionic surfactant vesicles that are hydrated immediately before use to produce aqueous niosome dispersions. To further enhance drug delivery, niosomes and proniosomes may be combined in the formulation.

[0341] Polymeric nanocarriers are generally formed from biodegradable polymers. Polymeric nanocarriers may be reservoir type (nanocapsules in which the compound is dissolved / distributed in the polymer core), matrix type (nanospheres in which the compound is entrapped in the polymer matrix), and combinations thereof. Polymeric nanocarriers may offer the benefits of low toxicity, easy modification (e.g., for targeted delivery), high drug loading capacity, small size, good water solubility, and biocompatibility. Exemplary polymeric nanocarriers for mitochondrial targeting include hydrophilic block polymers, e.g., polyethylene glycol (PEG), poly E-caprolactone (PCL). Other nanoparticles disclosed herein may be modified for mitochondrial targeting, e.g., by including surface hydrophilic block polymers (e.g., PEGylation). Polymersomes are artificial vesicles formed from amphiphilic self-assembling block copolymers. Polymersomes typically have a hydrophilic core and a lipophilic bilayer. Polymersomes are highly customizable. Drug encapsulation and release capabilities may be controlled by forming polymersomes with block copolymers that are biodegradable and / or responsive to stimuli. The composition and molecular weight of the polymersomes may be selected to control properties such as response to stimuli, membrane thickness, permeability, flexibility, and size (polymersomes may be designed to have radii from 50 nm to 5000 nm or larger). Polymersomes provide benefits such as improved skin elasticity and enhanced activation energy of skin cells.

[0342] Cubosomes are nanostructured particles formed from self-assembled liquid crystal particles of aqueous lipids and surfactants. Cubosomes are formed from a bicontinuous liquid phase that encloses two separate vesicles of aqueous formulations separated by a surfactant-controlled bilayer in a tightly packed structure. Cubosomes may be designed as honeycomb structures with more than two separate vesicles. The release of each vesicle may be controlled separately. Cubosomes may offer benefits such as providing controlled and / or targeted release of compounds, being lipid biodegradable, and having a high internal surface area with different drug loading modalities.

[0343] Micelles are colloidal aggregates that are generally amphiphilic in nature, with a hydrophilic head and a hydrophobic tail. The size and shape of the micelle nanoparticles may be selected by modifying the strength of the solution isotonicity, pH, temperature, and the nature of the amphiphilic molecules. Micelles may be utilized to improve the uptake of the compounds disclosed herein in mitochondria. For example, micelle formulations have been found to improve the bioavailability of poorly absorbed compounds, prevent mitochondrial swelling (indicating smaller mitochondrial permeability transition pore (mPTP) opening and prevention of injury), and protect cells from nitrosative stress, "Curcumin Micelles Improve Mitochondrial Function in Neuronal PC12 Cells and Brains of NMRI Mice - Impact on Bioavailability" (Hagl, 2015) (incorporated herein by reference in its entirety for all purposes). Micelles may be functionalized with targeting groups such as TPP, MPP, or PEGylation to target mitochondria. In some embodiments, the micelle nanoparticles may be polymeric micelles.

[0344] Dendrimers are hyperbranched macromolecules that can be formed from sugars, amino acids, and / or nucleotides. Dendrimers are generally formed from a central core, repeated branches, and various peripheral groups. The peripheral groups may be designed or functionalized for targeting applications such as targeted delivery. The benefits of dendrimers include the ability to provide drug encapsulation, high water solubility, high retention time, biodegradability, specificity, low toxicity, and surface modification capabilities that can provide properties such as monodispersity, multivalency, and stability. Dendrimers may be functionalized with targeting groups such as TPP, MPP, or PEGylation. Dendrimers may be formulated to encapsulate or conjugate with compounds.

[0345] Lipophilic cations are positively charged ions that can permeate plasma and mitochondrial membranes. Lipophilic cations tend to accumulate in mitochondria. Therefore, lipophilic cations such as TPP, dequalinum, and rhodamine 123 can be utilized for mitochondria-targeted therapeutic delivery. Delocalized lipophilic cations (DLCs) have strong mitochondria-targeting ability to cross membranes and drive specific aggregation of attached moieties within the mitochondria of cells. Therefore, DLCs can be utilized as nanoparticle carriers and / or surface modifications for targeted delivery. For example, the compositions disclosed herein can be conjugated to DLCs such as TPP, or encapsulated in another carrier with TPP surface functionalization for targeted delivery.

[0346] Solid-lipid nanoparticles are submicron colloidal carriers in the range of 50-1000 nm, e.g., 50-500 nm or 50-100 nm. SLNs are generally formed from physiological lipids interspersed in water or liquid surfactant solutions with an oil-based or lipoid core. SLNs may be prepared from complex glyceride mixtures, purified triglycerides, and waxes with phospholipid hydrophobic chains in a fatty matrix. SLNs offer benefits such as small size, large surface area, high drug loading capacity, and phase contact at the interface. SLNs may be designed to provide controlled or sustained release of compounds. In cosmetics and pharmaceuticals, SLNs may provide increased penetration of the compounds disclosed herein through the skin. SLNs may have ultraviolet (UV) resistance, occlusive properties that may be used to increase skin hydration, and good coalescence stability due to their solid nature, which reduces mobility and leakage of active molecules.

[0347] Nanostructured lipid carriers (NLCs) are a form of lipid nanoparticles formed by mixing solid lipids with spatially incompatible liquid lipid compositions to form an amorphous solid. NLCs may be of the incomplete, amorphous, or multilamellar type. NLC particles typically range in size from 10 nm to 1000 nm. When formulated from biodegradable and physiological lipids, NLCs exhibit very low toxicity. As such, NLC formulations may offer the benefits of reduced systemic side effects and higher drug loading capacity. NLCs may be designed to have a biphasic drug release pattern. For example, the first compound release profile may be immediate or controlled release, and the second compound release profile may be controlled or delayed release. Like SLNs, NLCs may also offer increased penetration of the compounds disclosed herein through the skin, ultraviolet (UV) resistance, occlusiveness that may be used to increase skin hydration, and good coalescence stability.

[0348] Nanoemulsions are kinetically and thermodynamically stable dispersions of lipids formed from oil and water phases combined with surfactants. Nanoemulsions disclosed herein may be oil-in-water, water-in-oil, or bicontinuous formulations. The properties of nanoemulsions may be engineered by controlling the method of preparation. Nanoemulsions are typically dispersed phases that contain small particles or droplets with low oil or water interfacial tension. Nanoemulsions are typically formed from a lipophilic core surrounded by a monolayer of phospholipids. Nanoemulsions offer benefits such as low viscosity, high kinetic stability, high interfacial area, high solubilization capacity, and increased rate of absorption. In cosmetics and pharmaceuticals, nanoemulsions may provide rapid penetration and active transport of active ingredients, as well as moisturization to the skin. Nanoemulsions may be formulated into foams, creams, sprays, or liquids.

[0349] Certain nanoparticle formulations, such as SLNs, nanoemulsions, liposomes, and niosomes, can be used in moisturizing formulations to provide moisturizers that retain moisture for extended periods of time.

[0350] Metal nanoparticles may be designed to have specific properties and may be in the shape of nanospheres, nanoshells, nanoclusters, nanorods, nanostars, nanocubes, branches, and nanotriangles. The shape, size, and dielectric properties of metal nanoparticles may have an effect on the resonant frequency. Metal nanoparticles may be designed to have high drug loading capacity and efficiently penetrate cell walls by controlling the size, surface area, and crystallinity. The benefits of metal nanoparticles include accelerating blood circulation, anti-inflammatory, antiseptic, improving skin firmness and elasticity, delaying aging, and energizing skin metabolism. Exemplary metal nanoparticles are gold, silver, and copper. Such metal nanoparticles have been shown to provide strong antifungal and / or antibacterial properties. Another exemplary metal nanoparticle is titanium dioxide (TiO2), which has been shown to provide protection from ultraviolet (UV) radiation. Metal nanoparticles may be designed to be inert, highly stable, biocompatible, and non-cytotoxic in nature. Two or more metals may be used to form metal nanocomposites with selected properties.

[0351] Nanospheres are spherical nanoparticles with a core-shell structure. Compounds can be encapsulated, conjugated, dissolved, or otherwise entrapped in nanoparticles. Nanospheres can be crystalline or amorphous in structure. Nanospheres can be biodegradable or non-biodegradable. Exemplary biodegradable biospheres include gelatin, modified starch, and albumin nanospheres. One exemplary non-biodegradable nanosphere is polylactic acid. In cosmetics and pharmaceuticals, nanospheres can be used to more precisely and efficiently deliver the compounds disclosed herein to deeper layers of the skin. Nanospheres have been shown to provide protection against actin aging.

[0352] Peptide-based nanomaterials are biomolecules made of several amino acids linked by peptide bonds. Peptides generally provide rapid clearance in the kidney due to enzymatic degradation. Peptide nanomaterials may offer several benefits including targeting and accumulation capabilities, small size, ease and customizability of production, and biocompatibility. Certain peptide nanomaterials may be designed to self-assemble into distinct shapes and sizes in response to environmental factors such as temperature, pH, ionic strength, or molecular interactions between the host and the peptide. Peptide nanoparticles may also be functionalized for targeted delivery. Functionalized peptides have been found to show improved targeting ability and enhanced efficacy.

[0353] One exemplary peptide nanoparticle with mitochondrial targeting capability is a mitochondrial penetrating peptide (MPP). MPP is a cell penetrating peptide that can efficiently penetrate the mitochondrial bilayer membrane. MPPs are generally designed or selected to be positively charged peptides. Due to the strong negative charge of the mitochondrial membrane, positively charged peptides can penetrate mitochondria. MMPs are described in more detail in "Mitochondrial targeted strategies and their application for cancer and other diseases treatment," (Li, 2020), which is incorporated by reference in its entirety for all purposes. As such, MPPs can be utilized as nanoparticle carriers and / or surface modifications for targeted delivery. For example, the compositions disclosed herein may be conjugated to MPP or encapsulated in another carrier with MPP surface functionalization for targeted delivery.

[0354] The formulations disclosed herein may include one or more skin permeation enhancers. In general, small and moderately lipophilic molecules may permeate the skin barrier. Other compounds may require a suitable skin permeation enhancer to permeate the barrier by either reducing the barrier properties of the skin or by actively driving the movement of the compound across the skin by inputting external energy. Skin permeation enhancers are described in "Penetration Enhancement of Topical Formulations," (Ng, 2018) and "Transdermal Delivery Systems in Cosmetics," (Kim, 2020), each of which is incorporated herein by reference in its entirety for all purposes.

[0355] The nanoparticle delivery carrier disclosed herein can provide enhanced skin permeation for compounds.The formulation disclosed herein may additionally or alternatively contain one or more chemical or physical skin permeation enhancers.Exemplary chemical skin permeation enhancers include alcohols such as ethanol and glycol, sulfoxides such as dimethyl sulfoxide, laurocapram, pyrrolidone, dimethyl isosorbide, isopropyl myristate, propylene glycol, oleic acid, eucalyptol, water / aqua (hydration), surfactants, urea, fatty acids, fatty alcohols, and terpenes and / or terpenoids.Exemplary physical skin permeation enhancers include rollers, scrapers, scrubbers, scrubbers, microdermabrasion needles, iontophoresis devices, electroporation devices, ultrasound devices such as sonophoresis, thermal ablation, magnetophoresis, photomechanical waves, electron beam irradiation, and low light therapy devices such as light emitting diode (LED) sources. Two or more skin permeation enhancers may be used synergistically in the formulation.

[0356] Chemical skin permeation enhancers may be included in dermatological, transdermal, cosmetic and pharmaceutical products to enhance the percutaneous absorption of drug compounds.Chemical enhancers can enable or improve the solubility, permeation and / or absorption of the compounds disclosed herein.Exemplary chemical enhancers are described below.

[0357] Water (aqua) generally increases the fluidity of the composition and provides higher permeability. In addition, water hydrates the skin barrier and modifies the skin lipids and / or proteins for improved permeation. Hydrating compounds, such as glycerol and urea, can facilitate transdermal penetration by facilitating hydration of the stratum corneum and forming hydrophilic diffusion channels in the barrier.

[0358] Alcohol solvents such as ethanol and propylene can provide enhanced permeation by increasing the fluidity of the compound and also act as good solvents. The degree of permeation may be selected by controlling the length of the alkyl chain of the fatty alcohol.

[0359] Surfactants generally solubilize lipophilic agents, including the active ingredients of the formulation and lipids in the stratum corneum. As such, surfactants may enhance skin permeability by partitioning into epithelial cell membranes and disrupting the packing of membrane lipids, forming structural defects that reduce membrane integrity. The effect of surfactants on skin permeation may be engineered by selecting the concentration and type of surfactant. Surfactants may be anionic, cationic, or nonionic. Anionic surfactants may be selected for skin or hair applications because they interact with keratin and lipids. Cationic surfactants may be selected for skin applications because they interact with skin proteins through polar interactions. Nonionic surfactants are generally less irritating to the skin and are well tolerated.

[0360] Fatty acids can increase transdermal drug absorption. Long-chain fatty acids, typically carboxylic acids with long, unbranched aliphatic tails, have been demonstrated to increase transdermal drug absorption as an effect of alkyl chain length. Low molecular weight alkanols can act as solubilizers to enhance the solubility of compounds in the fatty matrix of the stratum corneum. Polyunsaturated fatty acids, such as linoleic acid, alpha-linoleic acid, and arachidonic acid, can enhance skin penetration. One exemplary fatty acid chemical permeation enhancer is oleic acid. Oleic acid provides increased fluidity and reduced resistance to the penetration of molecules.

[0361] Terpenes are hydrocarbons commonly found in plant extracts. Terpenoids are terpenes that contain additional functional groups. One group of terpenes that can provide penetration enhancement is oxygen-containing terpenes. Exemplary oxygen-containing terpenes include menthol, thymol, carvacrol, menthone, and cineole. Such terpenes can enhance penetration in a similar mechanism to alcohol. However, terpenes can be considered natural products. The benefits of terpenes include high transdermal ability with minimal irritation and toxicity.

[0362] The compounds of the present disclosure may be formulated with mitochondrial targeting agents.Exemplary mitochondrial targeting agents include antibodies, polymeric moieties such as PEG, lipophilic cations such as triphenylphosphine (TPP), and peptides such as mitochondrial targeting peptides (MPP).

[0363] The compounds of the present disclosure may alternatively contain pharma- ceutically and / or cosmetically acceptable carrier substances necessary to approximate physiological conditions, such as, for example, pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, etc., such as, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc. For solid compositions, conventional non-toxic pharma-ceutically and / or cosmetically acceptable carriers can be used, including, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, magnesium carbonate, etc.

[0364] The composition of the present disclosure can also be formulated as liquid, microemulsion, or other ordered structure suitable for high concentration of active ingredient.Carrier can be solvent or dispersion medium, such as water, ethanol, polyol (such as glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof.The fluidity suitable for liquid can be maintained, for example, by using coating such as lecithin, by maintaining desired particle size in case of dispersible preparation, and by using surfactant.

[0365] In certain embodiments, the compound(s) and compositions of the present disclosure are administered in a time-release formulation, for example, in a composition that includes a delayed release polymer. Such compositions can be prepared with a carrier that protects against rapid release, for example, a controlled release medium, such as a polymer, a microencapsulated delivery system, or a bioadhesive gel. The extended delivery of various compositions of the present invention can be achieved by including in the composition an agent that delays absorption, for example, aluminum monosterate hydrogel and gelatin. When a controlled release formulation is desired, the controlled release binder suitable for use according to the present invention includes any biocompatible controlled release material that is inert to the active agent and can incorporate the bioactive agent. Many such materials are known in the art. Suitable formulations for topical administration include solutions, oils, creams, emulsions, and gels, which contain active ingredients and carriers known in the art.Topical formulations can be pharmaceutical or cosmetic formulations.In some embodiments, the compound is formulated as shampoo, conditioner, spray, cream, gel, balm, body wash, soap, lotion, or makeup.

[0366] The compounds and compositions of the present disclosure can be in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid excipient, such as water, for injection immediately prior to use. The appropriate unit dose, i.e., the effective amount, can be determined during appropriately designed clinical trials for each of the conditions in which the administration of the selected compound is indicated and, of course, varies depending on the desired clinical end point. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets. The requirements for an effective pharmacologic acceptable carrier for an injectable composition are well known to those skilled in the art. Pharmaceutics and Pharmacy Practice, JB Lippincott Co., Philadelphia, Pa., Banker and Chalmers, Eds., 238-250 (1982) and ASHP Handbook on Injectable Drugs, Toissel, 4 th ed., 622-630 (1986).

[0367] In addition, formulations suitable for rectal administration may be presented as suppositories by mixing with a variety of bases, such as emulsifying bases or water-soluble bases. Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate.

[0368] The compositions disclosed herein may be associated with various natural products, examples of which are provided below. The compositions disclosed herein may be formulated as natural products. The compositions disclosed herein may be administered in combination with natural products. The compositions disclosed herein may be incorporated into natural products. These natural products may be comprised of the formulations or compositions disclosed throughout this disclosure.

[0369] A natural product may be or include a product for commercial purposes, and may refer to dietary supplements and foods, such as foods, food supplements, medical foods, food additives, functional foods, or beverages, produced from natural sources. A natural product may have pharmacological or biological activity, which may be of therapeutic benefit, for example, in the treatment of a disease or condition. A natural product may be included in traditional medicine, cosmetic treatments, cosmetics, and spa treatments. A natural product referred to herein may include any one or more of the components described as a natural product incorporated into a composition or formulation that includes one or more other components, such as excipients. A preparation or formulation referred to as a natural product may include a natural product as defined herein, and one or more additional components or ingredients. Any of the compositions, preparations, or formulations discussed throughout this disclosure may be or include one or more natural products.

[0370] Those skilled in the art will recognize that suitable methods of administering the compounds of the present disclosure to a patient are available, and that while more than one route may be used to administer a particular compound, a particular route may provide a more immediate and more effective response than another route. EXAMPLES

[0371] The function and advantages of these and other embodiments can be better understood from the following examples, which are intended to be illustrative in nature and are not to be construed as limiting the scope of the invention.

[0372] Example 1 Identification of compounds from emblica extract To identify the compounds present in emblica, emblica extracts were prepared and analyzed by HPLC.

[0373] In this example, 40.9 grams of caplets containing E. officinalis extract (Himalaya Drug Company, Sugar Land, TX lot 112000924; each caplet contains 250 mg of fruit extract (45% tannin) and 350 mg of powdered stem (2% tannin)) were stirred with 250 ml of methanol at room temperature for 6 hours. The resulting solution was filtered to remove insoluble material, and the methanol layer was removed using a Rotovap (Fraction 1). The solid material resulting from the filtration step was stirred with an additional 250 ml of methanol at room temperature for 4 hours, filtered to remove insoluble material, and the filter funnel and methanol layer were removed using a Rotovap (Fraction 2). Fractions 1 and 2 were combined to give 8.25 grams of dark, hydrous solid.

[0374] 450 mg of the solid isolated as described above was dissolved in 2 ml of methanol. The resulting solution was sonicated until all contents were in solution and subjected to preparative HPLC purification using an ACCQPrep instrument (Teledyne ISCO, Lincoln, NE) with a Phenomenex Gemini® 5 μM NX-C18 110A 150×4.6 mm liquid chromatography column (Phenomenex, Torrance, CA). HPLC was performed using two solvent gradients as described in Table 3 below. Table 3: HPLC gradients [Table 3] Solvent A- Acetonitrile; Solvent B- Water Water was purified with a PureLab® Ultra water purification system (ELGA LabWater, Woodridge, IL).

[0375] The collected fractions were removed and the fractions were transferred to 11 separate vials. 1 mg of each fraction was taken and dissolved in 1 ml of methanol in a 1.5 ml vial. The resulting solution was sonicated until all contents were in solution and subjected to analytical HPLC analysis using an Agilent 1100 series instrument (Agilent Technologies, Santa Clara, CA) with a Phenomenex Gemini® 5 μM NX-C18 110A 150×4.6 mm liquid chromatography column (Phenomenex, Torrance, CA). HPLC was performed using two solvent gradients as described in Table 4 below. Table 4: HPLC gradients [Table 4] Solvent A- Acetonitrile with 0.1% trifluoroacetic acid (TFA) Solvent B- Water with 0.1% TFA Water was purified with a PureLab® Ultra water purification system (ELGA LabWater, Woodridge, IL).

[0376] To identify specific compounds in the A. emblica extract, standards shown in Table 5 were used. Table 5: emblica extract compound identification standards [Table 5]

[0377] Standards were prepared by dissolving 1 mg of standard in 1 ml of methanol, sonicating the solution, and adding the solution to a 2 ml vial. Samples were subjected to analytical HPLC as described above to verify purity. A combined standard solution was made by adding 100 ul of each standard to a separate 2 ml vial. Standards were used to identify the components in the emblica extract.

[0378] The following compounds (Table 6) were identified in the Emblica extract prepared and analyzed as described above. Table 6: Compounds identified in emblica extract [Table 6]

[0379] In addition, approximately 20 other compounds were present but have yet to be identified.

[0380] Example 2 Prophetic examples showing the in vivo energy and vitality improving effects of application of emblica extract Without further specific inclusion criteria, male and female subjects aged between 35 and 70 years showing aging in the form of prominent eye wrinkles are evaluated for the study. Subjects complete a baseline questionnaire of 10 multiple choice questions with predefined options to be selected. The questionnaire requests information on baseline energy levels and vitality. Other baseline parameters are measured for skin roughness (Ra, Rz) by DermaTOP (three-dimensional imaging of surface structures), skin hydration by Corneometer (Courage & Khazaka, Cologne, Germany) (capacitance measurement), and skin elasticity by Cutometer (optical measurement of skin displacement during suction of 300 mbar).

[0381] Subjects apply emblica extract (2-5 drops of emblica extract oil) or placebo topically to one side of the face twice daily for approximately 12 weeks. The study is performed in a split-face design. Anti-wrinkle properties are measured periorbitally in the area of ​​the crow's feet. Skin moisturizing effects are evaluated over the cheek bones. The effects of the emblica extract are compared to a reference product or placebo.

[0382] Subjects complete a final questionnaire at the end of the study. The final questionnaire requests information on baseline energy levels and vitality. Skin roughness (Ra, Rz), skin hydration, and skin elasticity are also measured at the end of the study. Baseline results are compared to final results.

[0383] Administration of the C. emblica extract is expected to improve energy levels and vitality. Administration of the C. emblica extract is also expected to improve aging-related parameters, including skin roughness, skin hydration, skin elasticity, and additional qualitative parameters measured by questionnaire.

[0384] Example 3 Restoration of mitochondrial DNA depletion and function and suppression of inflammation by Emblica extract in vivo In these experiments, the shaved dorsal skin of 8-9 week-old female C57BL / 6 control and mtDNA-depleted mice (expressing D1135A-POLG1) was topically treated daily with 200 μl of 50 mg / ml emblica extract ointment (prepared as described in the Methods section) or a corresponding amount of control ointment (lacking emblica extract) starting 1 week before the start of dox administration (200 mg / kg diet only) and continuing daily application for 16 weeks (112 days).

[0385] The effect of emblica extract treatment on reversal of mtDNA function was examined by staining paraffin-embedded dorsal skin sections of mtDNA-depleted mice treated with emblica extract ointment or control ointment with Oxphos complex IV antibody (COXII) (n=3). A statistically significant increase in COXII staining in mtDNA-depleted skin treated with emblica extract compared to control treatment was observed (Figure 1A). In addition, RT-PCR analysis of genes encoding mtDNA showed upregulation in mtDNA-depleted skin treated with emblica extract compared to control treatment (Figure 1B). Finally, an increase in mtDNA content was observed in skin samples from mtDNA-depleted mice treated with emblica extract compared to control treatment (Figure 1C).

[0386] As described herein, the skin of mtDNA-depleted mice showed an increase in mixed inflammatory cells in the dermis and around the adnexa, including mast cells, neutrophils and lymphocytes. After treatment with M. emblica extract, the skin of mtDNA-depleted mice showed a statistically significant decrease in inflammatory cells in the dermis and around the adnexa, including mast cells (Giemsa+ve positive cells; p=3.56E-07), granulocytes (MPO+ve cells; P=0.002), macrophages and histiocytes (CD163+ve cells; p=0.007), and B lymphocytes (Pax-5+ve cells; p=0.042) (Figure 1D). A decrease in the expression of inflammatory genes was also observed in skin samples from mtDNA-depleted mice treated with M. emblica extract versus those treated with control ointment (data not shown).

[0387] Example 4 Restoration of mitochondrial DNA depletion evidenced by reversal of wrinkled skin and hair loss by fucus extract in vivo To investigate the ability of other agents to restore mitochondrial DA and function, we investigated C. fucus extract. Reversal of mitochondrial DNA depletion is evidenced by reversal of wrinkled skin and hair loss. However, reversal of mitochondrial DNA depletion may have therapeutic effects on other indications.

[0388] In these experiments, the shaved dorsal skin of 8-9 week-old female C57BL / 6 control and mtDNA-depleted mice (expressing D1135A-POLG1) was topically treated daily with 200 μl of 50 mg / ml fucus extract ointment (prepared as described in the Methods section) or a corresponding amount of control ointment (lacking fucus extract) starting 1 week before the start of dox administration (200 mg / kg diet and 2 mg / mL in 5% sucrose water) and continued for 51 days (n=4 for each group).

[0389] The results are shown in Figure 2A. Consistent with previous results, dox administration to mtDNA-depleted mice resulted in significant hair loss and skin wrinkling phenotypes. Administration of fucus extract partially reversed the hair loss and skin wrinkling phenotypes compared to mtDNA-depleted mice without fucus extract treatment (dox administration alone). Control mice showed no effect when treated with dox alone or dox in combination with fucus extract.

[0390] Example 5 Restoration of mitochondrial DNA content by in vivo fucus extracts. The effect of fucus extract treatment on mitochondrial DNA content was also examined. Skin samples from animals in Example 4 were collected and analyzed for mitochondrial DNA content, and the results are shown in Figure 2B. As shown in Figure 2B, dox administration to mtDNA-depleted mice resulted in a significant decrease in mitochondrial DNA content compared to control mice. Administration of fucus extract completely restored mitochondrial DNA content. These results indicate that the beneficial effect of fucus extract on reversing the hair loss and wrinkled skin phenotype in mtDNA-depleted mice correlates with the preservation of mitochondrial DNA content.

[0391] Example 6 Expression of mitochondrial biogenesis regulatory proteins in vitro We tested C. emblica extract, its constituents, C. fucus extract, and related compounds in vitro for the induced expression of mitochondrial biogenesis regulatory proteins, including mitochondrial complex IV subunit 2 (COXII), mitochondrial transcription factor A (TFAM), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1a), at 6, 24, and 48 h after administration. The results are presented in the graphs in Figures 4A-4C and the gel electrophoresis images in Figure 5.

[0392] Many of the compositions tested showed increased protein expression compared to the DMSO control, which generally increased with increasing time.

[0393] Example 7 Prolonged expression of mitochondrial biogenesis regulatory proteins in vitro C. emblica extract was tested in vitro for induced expression of mitochondrial biogenesis regulatory proteins including mitochondrial complex IV subunit 2 (COXII), mitochondrial transcription factor A (TFAM), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1a) at 6, 12, 24, 48, 72, and 96 hours after administration, generally as described in the previous examples. Formulations of C. emblica extract at 0.01%, 0.05%, 0.1%, and 0.2% were tested. The results are presented in the graphs of Figures 6A-6F (COXII expression), 7A-7F (TFAM expression), and 8A-8F (PCG-1a expression). With a general trend, certain formulations induced higher expression after 96 hours than at earlier time points.

[0394] Emblica extracts formulated with nanoparticle delivery carriers were tested in vitro for induced expression of COXII at 6 and 24 hours after administration and compared to non-encapsulated compositions. The results are presented in the graphs of Figures 9A-9D. Specifically, Figure 9A shows COXII expression 6 hours after administration of the non-encapsulated composition. Figure 9B shows COXII expression 6 hours after administration of the nanoencapsulated composition. Figure 9C shows COXII expression 24 hours after administration of the non-encapsulated composition. Figure 9D shows COXII expression 24 hours after administration of the nanoencapsulated composition.

[0395] As shown in the data presented in Figures 9A-9D, all of the Emblica extract formulations showed an increase in COXII expression, with certain nanoencapsulated formulations showing a higher increase in COXII expression.

[0396] Example 8 Expression of mitochondrial biogenesis regulatory proteins in vitro Selected components of the C. emblica extract were tested in vitro for inducible expression of mitochondrial biogenesis regulatory proteins, including COXII and TFAM, at 6, 12, 24, and 48 hours post-administration, generally as described in the previous examples. Various concentrations of the formulations were tested. The results are presented in the graphs of Figures 10A through 22D. Briefly, chebulaginic acid, chebulinic acid, kaempferol, ellagic acid, ascorbic acid, citric acid, gallic acid, quercetin, and punicalagin were tested for inducible expression of target proteins at concentrations ranging from 2.5 μM to 100 μM.

[0397] Example 9 Nanoencapsulation of emblica extract and chebulinic acid C. emblica extract and chebulic acid formulations and nanoencapsulated chebulic acid formulations were tested in vitro for inducible expression of mitochondrial biogenesis regulatory proteins COXII and TFAM at 6, 12, 18, and 24 hours post-administration, generally as described in Example 4. Various concentrations of the formulations were tested. The results are presented in the graphs of Figures 23A-23D (COXII) and 24A-24D (TFAM).

[0398] The induction expression of COXII 6 hours after administration of various concentrations of nanoparticles loading 0.01%-0.2% of C. emblica extract and 50-400 μg / L was examined. The results are presented in the graphs of Figures 31A-31B. As shown in the graphs of Figures 25A-25B, higher efficacy was observed as a result of higher compound loading.

[0399] The induced expression of COXII and TFAM (with and without T. emblica extract) was examined 6, 12, 24, and 48 hours after administration of nanoparticle compositions with sizes ranging from 4.4 μm to 95 μm. The results are presented in the graphs of Figures 26A-26D (COXII), Figures 27A-27D (TFAM), and Figures 28A-28D (COXII). In the graphs, the legends of the x-axis are as follows: Nano A: Y100 (Emblica emblica extract) powder loaded (40% LF; average particle size = 11.6 μm) Nano B: Y100 (Emblica emblica extract) oil loaded (42% LF; average particle size = 9.4 μm) Nano C: Undoped blank (average particle size = 7.7 μm) Nano D: Large size octyl doped blank (average particle size = 95 μm) Nano E: Small octyl doped blank (average particle size = 4.4 μm)

[0400] As shown in the graphs of Figures 26A-26D, 27A-27D, and 28A-28D, greater efficacy was generally observed as a result of smaller particles.

[0401] The induced expression of COXII and TFAM 6, 12, 24, and 48 hours after administration of nanoparticle compositions with smaller sizes (with and without C. emblica extract) (with and without octyl doping) was examined. The results are presented in the graphs of Figures 29A-29D (COXII) and Figures 30A-30D (TFAM). As shown in the data presented in Figures 29A-29D and 30A-30D, the octyl doped particles showed higher expression than their non-doped counterparts.

[0402] Overall, the data indicate that the induction of mitochondrial biogenesis depends on the nanoparticle loading and nanoparticle size. Differences in nanoparticle preparation (between octyl doped and non-doped) and loading have an effect on the induction of mitochondrial biogenesis.

[0403] material and method Generation of mtDNA-depleted mice D1135A-POLG1 site-specific mutations were created in full-length human POLG1 complementary DNA (cDNA) using a site-directed mutagenesis kit (Agilent, Santa Clara, CA, USA). The primer sequences used for site-directed mutagenesis are as follows, with the mutation site in uppercase: D1135A_F: 5'-gcatcagcatccatgCGgaggttcgctacctgg-3' and D1135A_R: 5'-ccaggtagcgaacctcCGcatggatgctgatgc-3'. The mutations were confirmed by sequencing. D1135A-POLG1 cDNA was subcloned into the dox-inducible mammalian expression vector pTRE-Tight-BI-AcGFP1 (Clontech, Palo Alto, CA, USA). To obtain germline transmission of human D1135A-POLG1 (POLG1-DN), the pTRE-Tight-BI-AcGFP1-D1135A-POLG1 construct was microinjected into fertilized oocytes from C57BL / 6 mice. Potential founders were identified by screening genomic DNA from tail biopsies for the presence of the human Polg1 transgene using PCR. Heterozygous human POLG1 positive (+ / POLG1-DN + ) founder male mice were mated with CAG-rtTA3 (rtTA) C57BL / 6 female mice (Jackson Laboratories, stock no. 016532) to generate + / POLG1-DN + rtTA + Heterozygous transgenic mice were obtained. + rtTA + Heterozygous mice were interbred to generate homozygous POLG1-DN + rtTA + / POLG1-DN + rtTA + We generated mice (mtDNA-depleted mice) that showed normal litter size (6–7 pups) and Mendelian distribution of genotypes, i.e., wild-type, heterozygous + / POLG1-DN. + or + / rtTA +and homozygous POLG1-DN + rtTA + / POLG1-DN + rtTA + This resulted in a 1:2:1 distribution of POLG1-DN alleles, indicating that homozygosity for the POLG1-DN allele did not result in fetal or postnatal lethality. All mice received dox ad libitum in the diet (200 mg / kg diet) and water (2 mg / ml dox in 5% sucrose water). All animal experiments were performed by following the guidelines established by the Animal Care and Use Committee.

[0404] Histological and immunohistochemical analyses Skin from the dorsal and other tissues were fixed in buffered formalin, embedded in paraffin, sectioned (5 μM), and stained with hematoxylin and eosin. Skin sections were stained with Giemsa stain to detect mast cells, while MPO, CD3, CD163, and Pax-5 antibodies were used to detect other types of inflammatory cells by immunohistochemical analysis (Carson, et al., Histotechnology: A Self-Instruction Text, 3 ed., American Society for Clinical Pathology Press, Hong Kong, 2009).

[0405] RT-PCR and mtDNA content analysis To measure relative gene expression by RT-PCR, total cellular RNA from skin samples was isolated using Trizol (Invitrogen, Carlsbad, CA, USA). Approximately 1000-2000 ng of RNA was normalized across samples and cDNA was generated using an Iscript cDNA synthesis kit (Bio-Rad Laboratories, Hercules, CA, USA). The cDNA was then subjected to RT-PCR using Green Taq PCR mix (Promega, Madison, WI, USA) and gene-specific primers shown in Table 1 below. PCR products were run on 1.5-2% agarose gels and photographed using a gel photography system. At least three biological replicates were used in each PCR. β2-microglobulin or RNU6B were used as internal controls in each PCR.

[0406] Analysis of mtDNA content in skin and other tissues was performed as previously reported (Singh et al., PloS One, 10, e0139846, 2015). Briefly, mtDNA content was analyzed by real-time PCR with absolute quantification using the following primers: mMitoF: 5'-CTAGAAACCCCGAAACCAAA-3', mMitoR: 5'-CCAGCTATCACCAAGCTCGT-3', mB2MF: 5'-ATGGGAAGCCGAACATACTG-3', and mB2MR: 5'-CAGTCTCAGTGGGGGTGAAT-3'. Beta-2-microglobulin (B2M) was used as an internal control. [Table 7]

[0407] Microarray gene analysis Total RNA samples were extracted from MCF-7 cells expressing POLG1 D1135A 5 days after dox induction and from cells grown in the absence of dox for 5 days by Trizol extraction (Invitrogen). Illumina human microarray gene expression analysis was performed with the total RNA samples as previously described. BN-PAGE and Western blot analysis

[0408] Isolation of mitochondria was performed as previously described (Johnstone et al., J Biol Chem, 277, 42197-42204, 2002). To analyze mitochondrial OXPHOS supercomplexes, blue native polyacrylamide gel electrophoresis (BN-PAGE) was performed with mitochondrial fractions prepared from skin samples as previously described (Schagger et al., Mehtods Enzymol, 260, 190-202, 1995). Protein expression of mitochondrial OXPHOS subunits in skin samples was performed according to standard immunoblot. A premix cocktail containing primary monoclonal antibodies against subunits of OXPHOS complexes (Mitosciences, Eugene, OR, USA) was used to detect protein expression of OXPHOS supercomplexes in BN-PAGE analysis and OXPHOS subunits in immunoblot analysis. Voltage-dependent anion channel (VDAC) or β-actin antibodies were used as loading controls.

[0409] Analysis of enzymatic activities of OXPHOS complexes Isolated mitochondria were used for measuring the enzymatic activities of OXPHOS complexes as previously described (Owens et al., PloS One, 6, e23846, 2011).

[0410] Transmission electron microscopy Transmission electron microscopy of skin samples was performed as previously described (NAG et al., J Mol Cell Cardiol, 15, 301-317, 1983). Images were acquired using an FEI-Tecnai electron microscope.

[0411] cell culture Dermal fibroblasts from wild-type C57BL / 6 (control cells) and mtDNA-depleted mice containing the D1135A-POLG1 site-specific mutation (POLG1-DN cells) were generated and spontaneously immortalized as described (Todaro et al., J Cell Biol, 17, 299-313 (1963)). These cells were maintained in DMEM / F12 (Cellgro, Herndon, VA) supplemented with 10% FBS (Atlanta Biologicals, Lawrenceville, GA). To induce POLG1-DN expression in dermal fibroblasts, 1 μg / ml dox dissolved in water was added to the cells in culture, and after 6 days of incubation, the cells were washed with PBS and collected in Trizol for isolation of total RNA.

[0412] To estimate cell proliferation and cell survival, MTT assay was performed as previously described (Ronghe et al., J Steroid Biochem Mol Biol, 144 PtB, 500-512, 2014). Both control and POLG1-DN cells were first treated with dox (1 μg / ml) for 3 days, and then the cells were plated in 96-well plates at a density of 3000 cells / well with or without dox (1 μg / ml)-containing culture medium. Readings were taken every 24 hours.

[0413] Preparation of emblica extract Emblica officinalis was obtained from commercially available caplets (Himalaya Drug Company, Sugar Land, TX). Each caplet contains 600 mg (250 mg fruit extract (45% tannin), 350 mg powdered stem (2% tannin)). The caplets were crushed and dissolved in sterile water to make a 100 mg / ml solution, then filtered. The emblica solution was then mixed with a suitable ointment base for topical application. Suitable ointment bases include, but are not limited to, dermabase ointment (MARCELLE®; water, mineral oil, propylene glycol, stearyl alcohol, cetyl esters, cetyl alcohol, glyceryl stearate, sodium lauryl sulfate, lecithin, and methylparaben) and 1:1 w / v) and Geritrex hydrophilic ointment (NDC 54162-670-14). The ointment base and emblica solution may be added in any convenient ratio, for example, from 1:5 w / v emblica solution to ointment base to 5:1 w / v emblica solution to ointment base. The ointment base and emblica solution may be added in a ratio of 1:1 w / v emblica solution to ointment base.

[0414] Preparation of fucus extract Fucus vesiculosus (also known as bladder wrack, black tang, rockweed, bladder fucus, sea oak, cut weed, dyers fucus, red fucus, and rock wrack) powder was obtained from Maine Coast Sea Vegetables, Inc. (Hancock, MN). An aqueous solution (100 mg / ml) was prepared from the fucus powder and filtered. The fucus solution was then mixed with a suitable ointment base for topical application. Suitable ointment bases include, but are not limited to, dermabase ointment (MARCELLE®; water, mineral oil, propylene glycol, stearyl alcohol, cetyl esters, cetyl alcohol, glyceryl stearate, sodium lauryl sulfate, lecithin, and methylparaben) and 1:1 w / v) and Geritrex hydrophilic ointment (NDC 54162-670-14). The ointment base and the fucus solution may be added in any convenient ratio, for example, from 1:5 w / v fucus solution to ointment base to 5:1 w / v fucus solution to ointment base. The ointment base and the fucus solution may be added in a ratio of 1:1 w / v fucus solution to ointment base.

[0415] In vivo experimental design of emblica extract For animal experiments, the dorsal skin of mice is depilated (e.g., shaved under low-dose isoflurane inhalation anesthesia) approximately 2 days before the start of administration of composition to skin.Various active compositions and control compositions are topically applied to dorsal skin daily as described.The skin of mice is depilated before collection and analysis.

[0416] The emblica extract ointment, at a concentration of 100 mg / mL in water, was topically applied to a defined shaved area of ​​the dorsal skin of the mice on each day. The control composition contained the same amount of ointment base without the addition of emblica extract.

[0417] Fucus extract ointment at a concentration of 100 mg / mL aqueous solution was topically applied to a defined shaved area of ​​the dorsal skin of the mice on each day. The control composition contained the same amount of ointment base without the addition of fucus extract.

[0418] mtDNA-depleted mice (containing the D1135A-POLG1 mutation) and wild-type C57BL / 6 mice (control) were used in preventive and therapeutic experiments.

[0419] In the preventative experiment, daily emblica extract treatment as described above was started 7 days before starting dox-mediated induction of POLG1-DN. Both mtDNA-depleted and wild-type C57BL / 6 mice were divided into two treatment groups: i) control group (treated with ointment base only, n=5); and ii) test group (treated with emblica extract ointment, n=5). Daily administration was continued until the end of the experiment (day 112).

[0420] In the therapeutic experiment, wild-type C57BL / 6 mice (control) and mtDNA-depleted mice were first induced with dox for 30 days, and then the emblica extract treatment described above was applied daily until the end of the experiment (day 112). Both mtDNA-depleted and wild-type C57BL / 6 mice were divided into two treatment groups: i) control group (treated with ointment base only, n=5); and ii) test group (treated with emblica extract ointment, n=5).

[0421] During the period of dox administration, for Examples 1-13, mice were given dox ad libitum in the diet (200 mg / kg diet) and water (2 mg / ml dox in 5% sucrose water), and for Examples 14-15, mice were given dox ad libitum in the diet (2 mg / kg diet) and water (2 mg / ml dox in 5% sucrose water). A schematic of both the preventative and therapeutic in vivo experiments is shown in Figure 3, and it should be noted that in certain experiments of Examples 3-5, the time course of administration of emblica or fucus extracts continued beyond the end of the period specified in Figure 3 (i.e., up to 112 days).

[0422] statistical analysis Statistical analysis was performed using an unpaired Student's t-test. Data are presented as mean ± sem. A P value of <0.05 was considered significant. All cell experiments were repeated at least three times.

[0423] The phraseology and terminology used herein are for purposes of explanation and should not be considered as limiting. As used herein, the term "plurality" refers to two or more items or components. The terms "comprising," "including," "carrying," "having," "containing," and "involving," whether in the specification or claims, are open-ended terms, i.e., meaning "including but not limited to." Thus, the use of such terms is meant to encompass the items recited thereafter, and equivalents thereof, as well as additional items. With respect to the claims, only the transitional phrases "consisting of" and "consisting essentially of" are closed or semi-closed transitional phrases, respectively. The use of ordinal terms such as "first," "second," "third," etc. in the claims to modify claim elements does not in itself imply any priority, preference, or order of a claim element relative to an alternative or primary order in which the method acts are performed, but is merely used as a label to distinguish one claim element having a particular name from another element having the same name (in the absence of the use of ordinal terms) to identify the claim element.

[0424] Having thus described some aspects of at least one embodiment, it should be appreciated that various changes, modifications, and improvements will be readily apparent to those skilled in the art. Any feature described in any embodiment may be included in or substituted for any feature of any other embodiment. Such changes, modifications, and improvements are intended to be part of this disclosure and are intended to be within the scope of the present invention. Accordingly, the foregoing description and drawings are by way of example only.

[0425] Those skilled in the art should recognize that the parameters and configurations described herein are exemplary and that the actual parameters and / or configurations will depend on the particular application for which the disclosed methods and materials are used. Those skilled in the art should also recognize, or be able to ascertain, equivalents to the specific embodiments disclosed using no more than routine experimentation.

Claims

1. A composition comprising an effective amount of one or more emblica extracts, fucus extracts, and chebula extracts, or a pharmaceutically acceptable form thereof, or one or more compound components of one or more emblica extracts, fucus extracts, and chebula extracts, or a pharmaceutically acceptable form thereof, or one or more compounds having a similarity score of at least 95% to one or more compound components of emblica extracts, fucus extracts, and chebula extracts, or a pharmaceutically acceptable form thereof, for enhancing or improving the fertility of a subject, extending the reproductive lifespan of a subject, or reducing, preventing, or delaying the perimenopausal period, menopause, or the symptoms thereof of a subject.

2. A composition comprising an effective amount of one or more of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compound components of one or more of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compounds having a similarity score of at least 95% with one or more compound components of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, for use in a method for increasing the mitochondrial mass of an oocyte or preventing a decrease in the mitochondrial mass of an oocyte, or for improving embryonic development or improving the fertilization rate of an embryo, wherein the composition is administered to the subject related to the oocyte or the embryo or to the embryo.

3. A composition comprising an effective amount of one or more emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compound components of one or more emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compounds having a similarity score of at least 95% to one or more compound components of emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, for treating or preventing an ovarian disease or condition of interest.

4. A composition comprising an effective amount of one or more of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compound components of one or both of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, or one or more compounds having a similarity score of at least 95% to one or more compound components of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof, for use in a method for improving embryonic development or improving the fertilization rate of embryos, the method comprising administering the composition to at least one of a donor subject, a recipient subject, and the embryo, and transferring the oocyte cytoplasm to a recipient oocyte fertilized from a donor oocyte to form the embryo, wherein the donor oocyte has a higher mitochondrial DNA (mtDNA) content than the recipient oocyte.

5. The composition according to claim 1, wherein the emblica extract is derived from Emblica officinalis, the fucus extract is derived from Fucus vesiculosus, Fucus serratus, Fucus spiralis, or Fucus guiryi, or the chebula extract is derived from Terminalia chebula, Terminalia arborea, or Lumnitzera racemose.

6. The composition according to claim 1, wherein the compound component of the emblica extract, fucus extract, or chebula extract is benzoic acid substituted with 1 to 5 hydroxyl groups and optionally 1 to 3 O-(C1-C5 alkyl) groups or O-(C1-C5 alkenyl) groups, or a pharmaceutically acceptable form thereof, or benzene substituted with -CH=CH-(CH2)a-C(O)OH (wherein a is 0 to 5) and 1 to 5 hydroxyl groups, or a pharmaceutically acceptable form thereof, or a combination thereof.

7. The composition according to claim 1, wherein the compound component of the emblica extract, fucus extract, or chebula extract is gallic acid, vanillic acid, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, quercetin, fucoidan, emblicanin A, emblicanin B, punigluconin, pedankragine, punicaforin, filanemburin, kaempferol, ellagic acid, kevric acid, kebradic acid, punicalagin, any of the above metabolites, a compound having a similarity score of at least 95% with any of the above, or any of the above in a pharmaceutically acceptable form.

8. The composition according to claim 1, wherein one or more compound components of the emblica extract, fucus extract, or chebula extract, or one or more compound components having a similarity score of at least 95% to the compound components of the emblica extract, fucus extract, or chebula extract, are purified, for example, at least 80% purified, at least 85% purified, at least 90% purified, at least 95% purified, at least 98% purified, at least 99% purified, at least 99.9% purified, at least 99.99% purified, or at least 99.999% purified.

9. The composition according to claim 1, wherein the effective amount or therapeutic effective amount is sufficient to induce mitochondrial biogenesis and / or increase the expression of at least one protein selected from PGC-1a, TFAM, NRF-1, and COX II.

10. The composition according to claim 1, wherein the administration modulates the menstrual cycle of the subject or recipient, for example, normalizing the menstrual cycle, increasing ovulation events, and / or increasing anti-Müllerian hormone (AMH) levels.

11. The composition according to claim 2, wherein the administration reduces the probability of embryonic aneuploidy and / or Lie syndrome.

12. The composition according to claim 3, wherein the administration reduces or decreases the severity or frequency of at least one symptom of an ovarian disease or condition, such as dry skin or vagina, pelvic pain or muscle spasms, inflammation, prolonged or irregular menstrual cycles, and reduced ovulation events.

13. The composition according to claim 1, characterized in that the composition is administered topically, parenterally, for example, intravenously, intraperitoneally, intramuscularly, or enterally.

14. The composition according to claim 13, wherein the composition is formulated as a topical solution, an oil, a cream, an emulsion, or a gel, or as a parenteral liquid solution.

15. The composition according to claim 14, wherein the composition is formulated as a shampoo, conditioner, spray, cream, gel, ointment, body wash, soap, lotion, or makeup.

16. The composition according to claim 13, wherein the composition is formulated as an enteral capsule or tablet, or as a nutritional supplement or food, for example, a food, food supplement, medical food, food additive, functional food, or beverage.

17. The composition according to claim 1, wherein the composition is formulated for immediate release or sustained release, for example, controlled release or sustained release.

18. The composition according to claim 2, characterized in that the composition is administered in combination with one or more of clomiphene, tamoxifen, letrozole, metformin, gonadotropins, gonadotropin-releasing hormones, dopamine agonists, and other hormonal treatments, or a surgical procedure, such as fallopian tube surgery, laparoscopic surgery to remove or destroy cysts or submucosal fibroids, or laparoscopic ovarian drilling.

19. The composition according to claim 1, characterized in that the composition is administered in combination with a UV blocker, a moisturizer, a sunscreen, a wrinkle cream, a retinoid, an alpha-hydroxy acid, a beta-hydroxy acid, squalene, an antioxidant, tretinoin, glycosaminoglycans (GAGs), lactic acid, malic acid, citric acid, tartaric acid, hydroquinone, kojic acid, L-ascorbic acid, licorice extract, N-acetylglucosamine, niacinamide, soybeans, skin fillers or injectable drugs, such as hyaluronic acid or calcium hydroxyl apatite, botulinum toxin, laser resurfacing procedures, ultrasound therapy, chemical peels, such as glycolic acid peels, trichloroacetic acid or salicylic acid, or skin peeling procedures.

20. The composition according to claim 1, wherein the composition is administered in combination with or encapsulated in a biomaterial, selected from extracellular vesicles, collagen, hyaluronic acid, synthetic biomaterials (e.g., polylactic acid (PLA), polyglycolic acid (PGA), polycaprolactone (PCL), polyethylene glycol (PEG)), fibrin, alginate, and composite biomaterials.

21. The composition according to claim 3, wherein the ovarian disease or condition is selected from perimenopause, menopause, endometriosis, ovarian cysts, premenopausal or postmenopausal ovarian cancer, e.g., ovarian epithelial carcinoma, ovarian tumors, e.g., ovarian germ cell tumors, low-grade ovarian tumors, and ovarian stromal tumors, polycystic ovary syndrome (PCOS), primary ovarian insufficiency (POI), and ovarian torsion.

22. The composition according to claim 1, characterized in that the composition is administered in combination with one or more of the following: hormone therapy, for example, estrogen, progesterone, testosterone or a synthetic form thereof; selective serotonin reuptake inhibitors (SSRIs); gabapentin; clonidine; hormonal contraception; gonadotropin-releasing hormone (GnRH) agonists or antagonists; thermotherapy; pain relief; nonsteroidal anti-inflammatory drugs (NSAIDs), for example, ibuprofen or other analgesics; spironolactone; eflornithine; electrolysis; chemotherapy; radiotherapy; and surgery, for example, hysterectomy; bilateral salpingooophorectomy; and deburking surgery; and lifestyle changes, for example, weight loss; improved nutrition; and increased physical activity.

23. The composition according to claim 2, wherein the embryo is produced by in vitro fertilization (IVF).

24. The composition according to claim 2, wherein the method further comprises measuring the mitochondrial content of the embryo and selecting the embryo that responds to a measurement of mitochondrial content that is within a predetermined range.

25. The composition according to claim 4, wherein the oocyte transplant is complete, or the oocyte transplant is partial, for example by electrofusion or direct oocyte injection, or the transplant is performed by modified intracytoplasmic sperm injection (ICSI), autonomous germline mitochondrial energy transfer (AUGMENT) to the oocyte, nuclear genome transplant, for example, oocyte spindle transplant, oocyte vesicle (GV) transplant, pronuclear transplant (PNT), or polar body nuclear transplant (PBNT).

26. The composition according to claim 4, further comprising using CRISPR / Cas 9 gene editing technology on the mtDNA of the donor oocyte or the recipient oocyte.

27. ​​The method described above, To form the embryo by applying CRISPR / Cas 9 gene editing technology to the mitochondrial DNA (mtDNA) of the target oocyte to be fertilized; Introducing stem cell-derived mitochondrial DNA (mtDNA) into the target oocyte to be fertilized to form the embryo; or During development, an antioxidant is introduced into the culture medium of the embryo. The composition according to claim 2, further comprising:

28. The composition according to claim 1, characterized in that the composition comprises a nanoparticle delivery carrier, a skin permeability enhancer, or is administered in combination with a skin permeability enhancer, for example, a chemical skin permeability enhancer or a physical skin permeability enhancer, or a mitochondrial targeting agent, or a delivery carrier functionalized with a mitochondrial targeting agent.

29. The composition according to claim 1, wherein the compound component of the emblica extract, fucus extract, or chebula extract is derived from, purified from, or isolated from the emblica extract, fucus extract, or chebula extract, or is derived from, purified from, or isolated from, or synthesized from a source other than the emblica extract, fucus extract, or chebula extract.

30. A kit comprising: a preparation containing an effective amount of donor mtDNA in a therapeutically acceptable carrier; and a composition containing an effective amount of one or more of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof; or one or more compound components of one or more of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof; or one or more compounds having a similarity score of at least 95% to one or more compound components of the emblica extract, fucus extract, and chebula extract, or a pharmaceutically acceptable form thereof.

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