Topical compositions and methods for treating cutis laxa

Abstract

Disclosed herein are topical compositions comprising an extract of centella asiatica and mandelic acid, where the pH of the composition is less than 5, and methods of using the compositions to reduce cellular senescence in the skin and to treat cutis laxa. A topical composition comprising a centella asiatica extract and mandelic acid is effective in reducing the number of senescent cells in mammalian dermis, reducing senescence-related secretory phenotypes (SASP), down-regulating at least one SASP-related gene, increasing expression of at least one cell cycle-related gene in dermal fibroblasts, and reducing the number of senescent cells in mammalian dermis. At least one gene associated with skin inflammation is down-regulated and the fibroblast turnover rate is increased.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 442,424, filed January 31, 2023, pursuant to 35 USC §119(e), the entire contents of which are incorporated herein by reference. Background Technology

[0003] This disclosure generally relates to the field of topical compositions and methods of using them. In particular, this disclosure relates to topical compositions for treating and / or reducing the effects of cutisporosis. Cutisporosis is a term involving chronic skin dysfunction and fragility that leads to loss of function and ultimately the breakdown of the skin's protective mechanisms. People with cutisporosis exhibit atrophic, thinning skin that becomes fragile, manifesting as purpura and white pseudoscars on the extremities, with a tendency to tear and potentially leading to deep dissecting hematomas. See G. Kaya et al., “Dermatoporosis, a Prevalent Skin Condition Affecting the Elderly: Current Situation and Potential Treatments”, 37 *Clinics Dermatol.* 346–50 (2019). Progression of cutisporosis can lead to skin lacerations, delayed healing, bleeding, skin infections, and ultimately, medical emergencies.

[0004] Cutis laxa typically first appears between the ages of 40 and 60, and reaches full development between the ages of 70 and 90, potentially spanning a 50-year period. As the post-World War II "baby boomer" generation continues to age, and a higher proportion of this group transitions to the age of 65 or older, the incidence of cutis laxa is likely to increase. Fully developed cutis laxa is quite common in this group, with two studies in France setting the incidence at 32% and 37.5%. See J.H. Saurat et al., “A Simple Self-Diagnosis Tool to Assess the Prevalence of Dermatoporosis in France,” 31 *Journal of the European Association for Dermatology and Venereology*, 1380–86 (2017); V. Mengeaud et al., “Prevalence of Dermatoporosis in Elderly French Hospital In-Patients: A Cross-Sectional Study,” 166 *Br. J. Dermatol.*, 442–43 (2012). Therefore, improved compositions and methods for treating and / or reducing the effects of dermatoporosis will bring significant benefits. Summary of the Invention

[0005] In one aspect that can be combined with any other aspect or embodiment, this disclosure relates to a topical composition comprising: a Centella asiatica extract encapsulated in liposomes; and an alpha-hydroxy acid (AHA), wherein the composition has a pH less than 5. In some embodiments, the composition is in the form of an emulsion. In some embodiments, the emulsion is an oil-in-water (O / W) emulsion.

[0006] In some embodiments, the pH of the composition is greater than or equal to 4 and less than 5. In some embodiments, the Centella asiatica extract is present at a concentration of at least about 0.01 wt.% and less than 0.2 wt.% relative to the total weight of the composition. In some embodiments, the Centella asiatica extract is present at a concentration of about 0.01 wt.% to about 0.1 wt.% relative to the total weight of the composition.

[0007] In some embodiments, the Centella asiatica extract comprises a triterpene. In some embodiments, the triterpene is selected from one or more of the group consisting of: madecassoside, asiaticoside, madecassic acid, and asiatic acid. In some embodiments, the triterpene is present in the Centella asiatica extract at a concentration of at least about 70 wt.% relative to the total weight of the extract. In some embodiments, the triterpene is present in the composition at a concentration of at least about 0.005 wt.% and less than 0.2 wt.% relative to the total weight of the composition. In some embodiments, the triterpene is present in the Centella asiatica extract at a concentration of at least about 0.01 wt.% to about 0.05 wt.% relative to the total weight of the composition.

[0008] In some embodiments, the AHA comprises mandelic acid. In some embodiments, the AHA is present at a concentration of at least about 0.01 wt.% and less than 0.5 wt.% relative to the total weight of the composition. In some embodiments, the AHA is present at a concentration of about 0.05 wt.% to about 0.3 wt.% relative to the total weight of the composition.

[0009] In some embodiments, the liposomes are present at a concentration of about 0.03 wt.% to about 4 wt.% relative to the total weight of the composition. In some embodiments, the liposomes comprise lecithin, propylene glycol, or a combination thereof. In some embodiments, the liposomes comprise an antioxidant. In some embodiments, the antioxidant is tocopherol, tocopheryl acetate, or a combination thereof. In some embodiments, the average particle size of the liposomes is 100 nm to 300 nm. In some embodiments, the average particle size of the liposomes is about 250 nm.

[0010] In some embodiments, the Centella asiatica extract is present in the form of an active solution. In some embodiments, the liposomes and the active solution are present in a weight ratio of about 1:1 to about 1:20. In some embodiments, the liposomes and the active solution are present in a weight ratio of about 1:2 to about 1:10.

[0011] In some embodiments, the Centella asiatica extract and the AHA are present in a weight ratio of about 1:5 to 1:15.

[0012] In some embodiments, the topical composition further comprises an antioxidant, which is separate from any antioxidant in the liposomes. In some embodiments, the antioxidant separate from any antioxidant in the liposomes is tocopheryl acetate, tocopherol, or a combination thereof.

[0013] In some embodiments, the topical composition further comprises at least one of a moisturizer, emollient, or humectant. In some embodiments, the topical composition further comprises glycerin.

[0014] In some embodiments, the topical composition is a cream or emulsion.

[0015] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a topical composition comprising: Centella asiatica extract, wherein the Centella asiatica extract is encapsulated in liposomes; and an alpha-hydroxy acid (AHA), wherein: the composition is an emulsion; the pH of the composition is 4 to 5; and the composition reduces biomarkers of skin inflammation in mammalian skin cells.

[0016] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a topical composition comprising: an alpha-hydroxy acid (AHA); and a triterpenoid compound selected from the group consisting of asiaticoside, asiaticoside, asiaticopic acid, and asiatic acid, wherein the triterpenoid compound is encapsulated in liposomes, and wherein the pH of the composition is 4 to 5. In some embodiments, the AHA comprises mandelic acid.

[0017] In some embodiments, the AHA is present in the composition at a concentration of at least about 0.01 wt.% and less than 0.5 wt.% relative to the total weight of the composition.

[0018] In some embodiments, the triterpenoid compound is present in the composition at a concentration of at least about 0.005 wt.% and less than 0.2 wt.% relative to the total weight of the composition.

[0019] In some embodiments, the composition is an emulsion.

[0020] In some embodiments, the topical compositions disclosed herein can effectively reduce the number of senescent cells in the dermis of mammals.

[0021] In some embodiments, the topical compositions of this disclosure can effectively reduce the senescence-associated secretory phenotype (SASP) in mammalian epidermal cells.

[0022] In some embodiments, the local compositions of this disclosure can effectively downregulate at least one SASP-related gene selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0023] In some embodiments, the topical composition of this disclosure can effectively increase the expression of at least one cell cycle gene in mammalian dermal fibroblasts. In some embodiments, the at least one cell cycle gene in the mammalian fibroblasts is selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

[0024] In some embodiments, the topical compositions of this disclosure can effectively downregulate at least one gene associated with skin inflammation, said at least one gene associated with skin inflammation selected from the group consisting of: MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2.

[0025] In some embodiments, the local compositions of this disclosure can effectively downregulate at least one AP-1 complex transcription factor. In some embodiments, the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

[0026] In some embodiments, the local compositions of this disclosure can effectively activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

[0027] In some embodiments, in the local compositions disclosed herein, the Centella asiatica extract and the AHA are present at concentrations that can effectively synergistically downregulate at least one SASP-related gene selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0028] In some embodiments, in the local compositions disclosed herein, the Centella asiatica extract and the AHA are present at concentrations that can effectively and synergistically upregulate at least one cell cycle gene in mammalian dermal fibroblasts, the at least one cell cycle gene being selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

[0029] In some embodiments, in the local compositions disclosed herein, the Centella asiatica extract and the AHA are present at concentrations that can effectively synergistically downregulate at least one SASP-related gene selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0030] In some embodiments, in the local compositions disclosed herein, the Centella asiatica extract and the AHA are present at concentrations that can effectively and synergistically downregulate at least one AP-1 complex transcription factor. In some embodiments, the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

[0031] In some embodiments, in the local compositions disclosed herein, the Centella asiatica extract and the AHA are present at concentrations that can effectively and synergistically activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

[0032] In some embodiments, in the local compositions disclosed herein, the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically downregulate at least one SASP-related gene selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0033] In some embodiments, in the local compositions disclosed herein, the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically upregulate at least one cell cycle gene in mammalian dermal fibroblasts, the at least one cell cycle gene being selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

[0034] In some embodiments, in the local compositions disclosed herein, the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically downregulate at least one SASP-related gene selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0035] In some embodiments, in the local compositions disclosed herein, the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically downregulate at least one AP-1 complex transcription factor. In some embodiments, the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

[0036] In some embodiments, in the local compositions disclosed herein, the triterpenoid compound and the AHA are present at concentrations that can effectively and synergistically activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

[0037] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method of treating skin affected by cutis laxity, the method comprising: applying the topical composition of this disclosure to the affected skin.

[0038] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method for increasing fibroblast turnover in mammalian skin, the method comprising: applying to the mammalian skin a topical composition according to any one of claims 1 to 47.

[0039] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method for reducing the number of senescent cells in the dermis of a mammal, the method comprising: applying the topical composition of this disclosure to the mammalian skin.

[0040] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method for reducing the aging-associated secretory phenotype (SASP) in mammalian epidermal cells, the method comprising: applying a topical composition of the present disclosure to mammalian skin.

[0041] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method of downregulating at least one SASP-related gene in mammalian skin, the method comprising: applying a topical composition of the present disclosure to mammalian skin, wherein the at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

[0042] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method of increasing the expression of at least one cell cycle gene in mammalian dermal fibroblasts, the method comprising: applying a topical composition of the present disclosure to mammalian skin, wherein the at least one cell cycle gene in the mammalian fibroblasts is selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

[0043] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method of downregulating at least one gene associated with skin inflammation, the method comprising: applying a topical composition of the present disclosure to mammalian skin, wherein the at least one gene associated with skin inflammation is selected from the group consisting of MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2.

[0044] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method of downregulating at least one AP-1 complex transcription factor, the method comprising: applying a topical composition of the present disclosure to mammalian skin. In some embodiments, the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

[0045] In another aspect, which may be combined with any other aspect or embodiment, this disclosure relates to a method for activating cell cycle progression, the method comprising: applying a topical composition of the present disclosure to mammalian skin, wherein the composition upregulates CCNE2, CCNB1, or a combination thereof.

[0046] Further aspects and / or embodiments of the invention will be provided in the detailed description of the inventive technology shown below, but are not limited thereto. The following detailed description is exemplary and illustrative, but is not intended to be limiting. Attached Figure Description

[0047] The various objects, aspects, features, and advantages of this disclosure will become more apparent and better understood through a detailed description taken in conjunction with the accompanying drawings.

[0048] Figure 1A This is a small plot showing the fluorescence emission (590 nm) of Centella asiatica extract against human dermal fibroblasts relative to concentration in the alamar blue assay.

[0049] Figure 1B This is a small plot showing the fluorescence emission (590 nm) of Centella asiatica extract against human dermal keratinocytes relative to concentration in an Almar Blue assay.

[0050] Figure 1C This is a small plot showing the fluorescence emission (590 nm) of mandelic acid against human dermal fibroblasts in an Almar Blue assay, relative to its concentration.

[0051] Figure 1D This is a small plot showing the fluorescence emission (590 nm) of mandelic acid against human dermal keratinocytes relative to concentration in an Almar Blue assay.

[0052] Figure 2 This is a schematic diagram of biomarkers associated with aging of dermal cells.

[0053] Figure 3A This is a small diagram of IL8 gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0054] Figure 3B This is a small diagram of IL1B gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0055] Figure 3C This is a small diagram of HIST1H2BG gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0056] Figure 3DThis is a small diagram of UBE2C gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0057] Figure 4A This is a small graph showing the fold change in MMP2 gene expression in human keratinocytes that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0058] Figure 4B This is a small graph showing the fold change in HSPA8 gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0059] Figure 4C This is a small graph showing the fold change in CSF2RA gene expression in human keratinocytes that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0060] Figure 4D This is a small graph showing the fold change in CXCL1 gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0061] Figure 4E This is a small graph showing the fold change in IL2RB gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0062] Figure 4F This is a small graph showing the NFKBIA gene expression (fold change) in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0063] Figure 4G This is a small graph showing the fold change in PTGS2 gene expression in human keratinized cells that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0064] Figure 5 This is a small graph showing the fold change in CDKN2B gene expression in human keratinocytes that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0065] Figure 6This is a small graph showing the fold change in gene expression of cell cycle-related genes in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0066] Figure 7A This is a small graph showing the fold change in WEE1 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0067] Figure 7B This is a small graph showing the fold change in H2AFX gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0068] Figure 7C This is a small graph showing the fold change in LMNB1 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0069] Figure 7D This is a small graph showing the fold change in CCNB2 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0070] Figure 7E This is a small graph showing the fold change in BUB1 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0071] Figure 7F This is a small graph showing the fold change in CDC45 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0072] Figure 7G This is a small graph showing the fold change in CDK1 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0073] Figure 7H This is a small graph showing the fold change in BIRC5 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0074] Figure 7IThis is a small graph showing the fold change in MCM10 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0075] Figure 7J This is a small graph showing the fold change in CCNE2 gene expression in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing Centella asiatica extract, mandelic acid, or a combination of Centella asiatica extract and mandelic acid.

[0076] Figure 8 This is a small plot showing the gene expression (fold change relative to the untreated group) of JunB and related proteins in human dermal fibroblasts that have grown to aggregate and been treated with a composition containing a combination of Centella asiatica extract and mandelic acid.

[0077] Figure 9A This image shows a p16-stained (yellow) human skin sample treated with the topical composition according to this disclosure, compared to an untreated cell sample. Cell nuclei are shown in blue. The circled areas show the presence of senescent cells in the dermis (untreated) and the migration of senescent cells into the epidermis (test composition).

[0078] Figure 9B A p16-stained (yellow) human skin sample treated with the topical composition according to this disclosure is shown compared to an untreated cell sample. Cell nuclei are shown in blue. Circled areas show the presence of senescent cells in the dermis (untreated) and the migration of senescent cells to the epidermis (test composition). The senescent cells in the center of the image of the untreated sample are present in hair follicles, where cells undergo natural senescence as part of the hair cycle.

[0079] Figure 9C A p16-stained (yellow) human skin sample treated with the topical composition according to this disclosure is shown compared to an untreated cell sample. Cell nuclei are shown in blue. The circled areas show the presence of senescent cells in the dermis (untreated) and the migration of senescent cells to the epidermis (test composition). The senescent cells on the right side of the image of the untreated sample and the senescent cells from the lower left corner to the center of the image of the treated sample are present in hair follicles, where cells undergo natural senescence as part of the hair cycle.

[0080] Figure 9DThe image shows a p16-stained (yellow) human skin sample treated with the topical composition according to this disclosure, compared to an untreated cell sample. Cell nuclei are shown in blue. Circled areas show the presence of senescent cells in the dermis (untreated) and the migration of senescent cells towards the epidermis (test composition). The senescent cells at the bottom of the image of the untreated sample and the senescent cells in the middle to upper portion of the image of the treated sample are present in hair follicles, where cells undergo natural senescence as part of the hair cycle. Detailed Implementation

[0081] In the following description, numerous specific details are set forth to provide a thorough understanding of the invention. Certain exemplary embodiments of the invention may be practiced without some or all of these specific details. In other instances, well-known process operations have not been described in detail to avoid unnecessarily obscuring the invention.

[0082] Topical Composition

[0083] Centella Asiatica extract

[0084] The topical composition according to this disclosure comprises Centella asiatica extract. Centella asiatica is a traditional herb widely used in Asia and increasingly popular in Western countries. Centella asiatica extract is used as a natural remedy and has wide applications in wound healing, skin barrier enhancement, skin hydration, reducing wrinkles, and alleviating skin conditions such as acne, atopic dermatitis, and burns. Its use in wound healing is attributed to its activity in improving collagen synthesis and microcirculation. It is also believed to regulate cellular aging and possesses anti-hyaluronidase activity. For general reference, see, for example, E. Arribas-Lopez et al., “A systematic review of the effect of Centella Asiatica on Wound Healing”, 19 *Int'l J. Envtl. Res. Pub. Health* 3266 (2022); S. Saeidinia et al., “Partial Thickness Burn Wound Healing by Topical Treatment: A Randomized Controlled Comparison Between Silver Sulfadiazine and Centiderm”, 96 *Medicine* 103 (2017); NKNema et al., “Matrix Metalloproteinase, Hyaluronidase and Elastase Inhibitory Potential of Standardized Extract of Centella Asiatica”. Asiatica, “Centella Asiatica Extracts Modulate HydrogenPeroxide-Induced Senescence in Human Dermal Fibroblasts”, 51 Pharm.Biol. 1182–87 (2013); YJ Kim et al., “Centella Asiatica Extracts Modulate HydrogenPeroxide-Induced Senescence in Human Dermal Fibroblasts”, 20 Experimental Dermatology, 998–1003 (2011).

[0085] Centella asiatica extract is available in the form of an aqueous or organic extract, and its therapeutic properties are attributed to the presence of triterpenoids: asiaticoside, asiaticoside, asiaticopic acid, and asiatic acid. Centella asiatica extract can contain high concentrations of triterpenoids. For example, commercially available Centella asiatica extracts (e.g., HETEROSIDES, available from Seppic) can contain more than 70 wt.% triterpenoids. In some embodiments, Centella asiatica extract contains HETEROSIDES (Seppic). In some embodiments, Centella asiatica extract contains triterpenoids selected from the group consisting of: asiaticoside, asiaticoside, asiaticopic acid, and asiatic acid.

[0086] Centella asiatica extract contains triterpenoids at concentrations suitable for achieving therapeutic benefits (e.g., reducing cellular aging). In some embodiments, triterpenoids are present in the Centella asiatica extract at concentrations greater than or equal to about 5 wt.%, greater than or equal to about 8 wt.%, greater than or equal to about 10 wt.%, greater than or equal to about 15 wt.%, greater than or equal to about 20 wt.%, greater than or equal to about 25 wt.%, greater than or equal to about 30 wt.%, greater than or equal to about 35 wt.%, greater than or equal to about 40 wt.%, greater than or equal to about 45 wt.%, greater than or equal to about 50 wt.%, greater than or equal to about 55 wt.%, greater than or equal to about 60 wt.%, greater than or equal to about 65 wt.%, greater than or equal to about 70 wt.%, greater than or equal to about 75 wt.%, greater than or equal to about 80 wt.%, greater than or equal to about 85 wt.%, greater than or equal to about 90 wt.%, greater than or equal to about 95 wt.%, or any range or value thereof relative to the total weight of the Centella asiatica extract. In some embodiments, the Centella asiatica extract comprises a triterpenoid compound at a total weight concentration of 70 wt.% or greater relative to the total weight of the Centella asiatica extract.

[0087] Centella asiatica extract (including triterpenoids and any solvent, including water) is present in the topical composition at any suitable concentration suitable for achieving therapeutic benefits (e.g., reducing cellular senescence). In some embodiments, Centella asiatica extract is present in the topical composition at a concentration of at least about 0.01 wt.%, at least about 0.02 wt.%, at least about 0.03 wt.%, at least about 0.04 wt.%, at least about 0.05 wt.%, at least about 0.06 wt.%, at least about 0.07 wt.%, at least about 0.08 wt.%, at least about 0.09 wt.%, at least about 0.10 wt.%, at least about 0.15 wt.%, at least about 0.20 wt.%, at least about 0.25 wt.%, at least about 0.30 wt.%, at least about 0.35 wt.%, at least about 0.40 wt.%, at least about 0.45 wt.%, at least about 0.50 wt.%, or any range or value between thereof.

[0088] In some embodiments, Centella asiatica extract is present in the topical composition at a concentration of less than or equal to about 0.5 wt.%, less than or equal to about 0.45 wt.%, less than or equal to about 0.40 wt.%, less than or equal to about 0.35 wt.%, less than or equal to about 0.30 wt.%, less than or equal to about 0.25 wt.%, less than or equal to about 0.20 wt.%, less than or equal to about 0.15 wt.%, less than or equal to about 0.10 wt.%, less than or equal to about 0.09 wt.%, less than or equal to about 0.08 wt.%, less than or equal to about 0.07 wt.%, less than or equal to about 0.06 wt.%, less than or equal to about 0.05 wt.%, or any range or value between thereof, relative to the total weight of the topical composition.

[0089] In some embodiments, Centella asiatica extract is present in the topical composition at a concentration of at least about 0.01 wt.% and less than 0.2 wt.%, about 0.01 wt.% to about 0.15 wt.%, about 0.01 wt.% to about 0.10 wt.%, about 0.01 wt.% to about 0.08 wt.%, about 0.01 wt.% to about 0.05 wt.%, or any range or value thereof, relative to the total weight of the topical composition.

[0090] The examples provided above include illustrative concentrations of Centella asiatica extract based on high concentrations of triterpenoids (e.g., greater than 70% triterpenoids available from Secico). In examples where a lower amount of triterpenoids is present in the Centella asiatica extract, the concentration of the Centella asiatica extract in the illustrative examples can be increased to ensure that a minimum amount of the triterpenoid concentrate disclosed herein is present in the formulation.

[0091] In some embodiments, the Centella asiatica extract is present in the form of an active solution. As used herein, the term "active solution" refers to a solution containing Centella asiatica extract (including any terpenoid compounds) and a solvent. In some embodiments, the Centella asiatica extract is present in the composition at a concentration lower than the recommended concentration for formulating the Centella asiatica extract into a topical composition (e.g., a recommended concentration of about 0.2 wt.% to 0.5 wt.%). Thus, in some embodiments, the Centella asiatica extract is present in the form of an active solution at a concentration of 0.01 wt.% to less than 0.2 wt.% (e.g., 0.01 wt.% to 0.15 wt.%, 0.01 wt.% to 0.10 wt.%, 0.01 wt.% to 0.08 wt.%, 0.01 wt.% to 0.05 wt.%) relative to the total weight of the composition. The inventors have surprisingly discovered that low concentrations (e.g., less than 0.2 wt.%) of the Centella asiatica extract have numerous therapeutic benefits, which are discussed below (see Method of Use). In some embodiments, low concentrations of Centella asiatica extract, together with low concentrations of alpha-hydroxy acids (AHAs), achieve synergistic therapeutic activity.

[0092] Given the concentration range of the Centella asiatica extract in the total composition (e.g., at least about 0.01 wt.% and less than 0.2 wt.%) and the concentration of triterpenoids in the Centella asiatica extract relative to the total weight of the Centella asiatica extract (e.g., at least about 5 wt.% to about 95 wt.%), it is evident that, whether or not the triterpenoids are part of the Centella asiatica extract, they can be present at concentrations suitable for achieving therapeutic benefits (e.g., reducing cellular senescence). In some embodiments, the triterpenoid compound may be present in amounts of at least about 0.0005 wt.%, at least about 0.001 wt.%, at least about 0.002 wt.%, at least about 0.003 wt.%, at least about 0.004 wt.%, at least about 0.005 wt.%, at least about 0.006 wt.%, at least about 0.007 wt.%, at least about 0.008 wt.%, at least about 0.009 wt.%, at least about 0.01 wt.%, at least about 0.02 wt.%, at least about 0.03 wt.%, at least about 0.04 wt.% of the total weight of the topical composition, by weight. The concentration is present in the topical composition at least about 0.05 wt.%, at least about 0.06 wt.%, at least about 0.07 wt.%, at least about 0.08 wt.%, at least about 0.09 wt.%, at least about 0.10 wt.%, at least about 0.11 wt.%, at least about 0.12 wt.%, at least about 0.13 wt.%, at least about 0.14 wt.%, at least about 0.15 wt.%, at least about 0.16 wt.%, at least about 0.17 wt.%, at least about 0.18 wt.%, at least about 0.19 wt.%, or any value range between therewith.

[0093] In some embodiments, the triterpenoid compound may be present in amounts less than about 0.2 wt.%, less than or equal to about 0.19 wt.%, less than or equal to about 0.18 wt.%, less than or equal to about 0.17 wt.%, less than or equal to about 0.16 wt.%, less than or equal to about 0.15 wt.%, less than or equal to about 0.14 wt.%, less than or equal to about 0.13 wt.%, less than or equal to about 0.12 wt.%, less than or equal to about 0.11 wt.%, less than or equal to about 0.10 wt.%, or less than or equal to about 0.09 wt.% by weight relative to the total weight of the topical composition. The concentration of any range or value between 0.08 wt.% and 0.09 wt.% is present in the topical composition.

[0094] In some embodiments, the triterpenoid compound may be present in the topical composition at a concentration of at least about 0.0005 wt.% and less than 0.2 wt.%, 0.001 wt.% and less than 0.2 wt.%, about 0.001 wt.% to about 0.15 wt.%, about 0.001 wt.% to about 0.1 wt.%, about 0.001 wt.% to about 0.05 wt.%, about 0.001 wt.% to about 0.01 wt.%, at least about 0.005 wt.% and less than 0.2 wt.%, about 0.005 wt.% to about 0.15 wt.%, about 0.005 wt.% to about 0.10 wt.%, about 0.005 wt.% to about 0.05 wt.%, about 0.005 wt.%, to about 0.01 wt.%, or any range or value thereof, relative to the total weight of the topical composition.

[0095] Liposomes

[0096] The triterpenoids present in Centella asiatica extract have large molecular weights and are chemically unstable, which can inhibit skin adsorption after topical application. Therefore, in some embodiments, Centella asiatica extract (and the triterpenoids therein) is encapsulated within a delivery medium (e.g., liposomes, lipid nanoparticles, or combinations thereof) to improve the stability of the triterpenoids and enhance the delivery of Centella asiatica extract to the skin by improving penetration into the epidermis and dermis. In some embodiments, Centella asiatica extract is encapsulated in liposomes. Liposome compositions can improve the distribution, efficacy, bioavailability, and / or activity of the active ingredient by improving delivery and tissue (e.g., skin) penetration. In some instances, the improvement in delivery and skin penetration is due to the encapsulation of the active ingredient in liposomes.

[0097] Liposomes encapsulating Centella asiatica extract can be prepared using lecithin and other phospholipids. Liposomes form when phospholipids such as lecithin are placed in water and thus form a bilayer or a series of bilayers; each bilayer is separated by water molecules once sufficient energy is provided. Liposomes can be formed by sonicating phospholipids in water. Low shear rates produce multilayered liposomes. Continuous high-shear sonication tends to form smaller monolayer liposomes. Hydrophobic compounds (e.g., triterpenoids, antioxidants such as tocopherol and / or tocopheryl acetate, and oils such as sunflower seed oil) can dissolve into the phospholipid bilayer membrane. Liposomes deliver Centella asiatica extract in lipid bilayers as described herein.

[0098] The phase transition temperature of the phospholipids used to prepare the liposomes described herein can be from about 10°C to about 25°C. In some embodiments, the phase transition temperature of the phospholipids is about 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, 32°C, 34°C, 36°C, 38°C, 40°C, or greater than 40°C. In some embodiments, the phase transition temperature of the phospholipids is in the range of about 10°C to about 40°C, about 12°C to about 36°C, about 14°C to about 32°C, about 16°C to about 20°C, or about 21°C to about 25°C.

[0099] In some embodiments, the liposomes encapsulating Centella asiatica extract are prepared by a method comprising: combining an "active solution" to form a mixture, said active solution being a solution containing Centella asiatica extract and a solvent (e.g., water); and contacting said active solution with an aqueous solution containing the liposomes. In some instances, the contact occurs at a temperature between about 10°C and about 25°C. In some instances, the contact occurs at temperatures of about 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, 32°C, 34°C, 36°C, 38°C, 40°C, or greater than 40°C. In some instances, the contact occurs at temperatures in the range of about 10°C to about 40°C, about 12°C to about 36°C, about 14°C to about 32°C, about 16°C to about 20°C, or about 21°C to about 25°C.

[0100] In some embodiments, the solvent is water. In some embodiments, the solvent is an organic solvent. Exemplary organic solvents include, but are not limited to, petroleum ether, cyclohexane, toluene, carbon tetrachloride, dichloromethane, chloroform, diethyl ether, diisopropyl ether, ethyl acetate, butanol, n-propanol, ethanol, methanol, polyethylene glycol, propylene glycol, and pyridine. In some instances, the solvent is a glycol. In some instances, the solvent is butanediol. In some instances, the solvent is octyl ethylene glycol. In some instances, the solvent is propylene glycol.

[0101] Solvents can be used at any suitable concentration to form stable liposomes. In some embodiments, the solvent (e.g., propylene glycol) is present in amounts greater than or equal to about 0.001 wt.%, greater than or equal to about 0.005 wt.%, greater than or equal to about 0.01 wt.%, greater than or equal to about 0.02 wt.%, greater than or equal to about 0.05 wt.%, greater than or equal to about 0.10 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, or greater than or equal to about 2.0 wt.% of the total weight of the liposomes. Concentrations of 0.0%, greater than or equal to about 2.5 wt.%, greater than or equal to about 3.0 wt.%, greater than or equal to about 3.5 wt.%, greater than or equal to about 4.0 wt.%, greater than or equal to about 4.5 wt.%, greater than or equal to about 5.0 wt.%, greater than or equal to about 5.5 wt.%, greater than or equal to about 6.0 wt.%, greater than or equal to about 6.5 wt.%, greater than or equal to about 7.0 wt.%, greater than or equal to about 8 wt.%, greater than or equal to about 9 wt.%, greater than or equal to about 10 wt.%, or any range or value between are provided. In some embodiments, the solvent is selected from propylene glycol, butanediol, octyl ethylene glycol, or combinations thereof. In some embodiments, the solvent is propylene glycol.

[0102] In some embodiments, forming liposomes encapsulating Centella asiatica extract comprises combining Centella asiatica extract and a solvent (e.g., water) to form an active solution; and contacting the active solution with an aqueous solution containing liposomes, wherein the aqueous solution contains water and liposomes at any concentration suitable for forming stable liposomes and encapsulating the Centella asiatica extract in the liposomes. In some embodiments, the aqueous solution contains water at a concentration greater than or equal to about 20 wt.%, 30 wt.%, 40 wt.%, 50 wt.%, 60 wt.%, 70 wt.%, 80 wt.%, or 90 wt.% relative to the total weight of liposomes, water, and Centella asiatica extract. In some embodiments, the aqueous solution contains water in the range of about 10 wt.% to about 95 wt.%, about 20 wt.% to about 90 wt.%, about 30 wt.% to about 85 wt.%, about 40 wt.% to about 80 wt.%, or about 50 wt.% to about 60 wt.% relative to the total weight of liposomes, water, and Centella asiatica extract.

[0103] In some embodiments, the aqueous solution comprises liposomes at a weight concentration of at least or about 10 wt.%, at least about 20 wt.%, at least about 30 wt.%, at least about 40 wt.%, at least about 50 wt.%, at least about 60 wt.%, or any range or value between therewith, relative to the total weight of liposomes, water, and Centella asiatica extract. In some embodiments, the aqueous solution comprises liposomes at a weight concentration of about 10 wt.% to about 80 wt.%, about 20 wt.% to about 70 wt.%, or about 30 wt.% to about 60 wt.%, relative to the total weight of liposomes, water, and Centella asiatica extract. The liposome to water ratio (w / w) can be in the range of about 1:9 to about 3:7. In some embodiments, the ratio (w / w) of liposomes to water may be greater than or equal to about 1:10, greater than or equal to about 1:9, greater than or equal to about 1:8, greater than or equal to about 1:7, greater than or equal to about 1:6, greater than or equal to about 1:5, greater than or equal to about 1:4, greater than or equal to about 1:3, or greater than or equal to about 1:2.

[0104] The method for producing liposomes encapsulating Centella asiatica extract can produce an encapsulation efficiency of no more than 100%. In some embodiments, the encapsulation efficiency is no more than 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5%.

[0105] Liposomes can be present in the topical composition at any suitable concentration to improve the delivery of Centella asiatica extract to the extracellular matrix of the epidermis or dermis of the skin. In some embodiments, the liposomes are present in amounts greater than or equal to about 0.01 wt.%, greater than or equal to about 0.02 wt.%, greater than or equal to about 0.03 wt.%, greater than or equal to about 0.04 wt.%, greater than or equal to about 0.05 wt.%, greater than or equal to about 0.06 wt.%, greater than or equal to about 0.07 wt.%, greater than or equal to about 0.08 wt.%, greater than or equal to about 0.09 wt.%, greater than or equal to about 0.10 wt.%, greater than or equal to about 0.15 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.30 wt.%, or greater than or equal to about 0.02 wt.%, respectively, relative to the total weight of the composition. The concentration of 0.35 wt.%, greater than or equal to about 0.40 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.6 wt.%, greater than or equal to about 0.7 wt.%, greater than or equal to about 0.8 wt.%, greater than or equal to about 0.9 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about 2.0 wt.%, greater than or equal to about 2.5 wt.%, greater than or equal to about 3.0 wt.%, greater than or equal to about 3.5 wt.%, greater than or equal to about 4.0 wt.%, greater than or equal to about 4.5 wt.%, greater than or equal to about 5.0 wt.%, or any range or value between these values, is present in the topical composition.

[0106] In some embodiments, liposomes are present in the topical composition at concentrations of about 0.01 wt.% to about 5 wt.%, about 0.05 wt.% to about 5 wt.%, about 0.1 wt.% to about 5 wt.%, about 0.03 wt.% to about 4 wt.%, about 0.05 wt.% to about 4 wt.%, about 0.05 wt.% to about 3 wt.%, about 0.05 wt.% to about 2 wt.%, about 0.05 wt.% to about 1 wt.%, about 0.05 wt.% to about 0.5 wt.%, about 0.05 wt.% to about 0.1 wt.%, about 0.1 wt.% to about 2 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.2 wt.% to about 0.8 wt.%, about 0.3 wt.% to about 0.7 wt.%, or any range or value between thereof, relative to the total weight of the composition.

[0107] In some embodiments, the liposomes have any size suitable for improving the delivery of Centella asiatica extract to the skin. In some embodiments, the average diameter of the liposomes is greater than or equal to about 50 nm, greater than or equal to about 60 nm, greater than or equal to about 70 nm, greater than or equal to about 80 nm, greater than or equal to about 90 nm, greater than or equal to about 100 nm, greater than or equal to about 110 nm, greater than or equal to about 120 nm, greater than or equal to about 130 nm, greater than or equal to about 140 nm, greater than or equal to about 150 nm, greater than or equal to about 160 nm, greater than or equal to about 170 nm, greater than or equal to about 180 nm, greater than or equal to about 190 nm, greater than or equal to about 200 nm, greater than or equal to about 210 nm, greater than or equal to about 220 nm, greater than or equal to about 230 nm, greater than or equal to about 240 nm, greater than or equal to about 2 ...50 nm, greater than or equal to about 160 nm, greater than or equal to about 170 nm, greater than or equal to about 180 nm, greater than or 260nm, greater than or equal to about 270nm, greater than or equal to about 280nm, greater than or equal to about 290nm, greater than or equal to about 300nm, greater than or equal to about 310nm, greater than or equal to about 320nm, greater than or equal to about 330nm, greater than or equal to about 340nm, greater than or equal to about 350nm, greater than or equal to about 360nm, greater than or equal to about 370nm, greater than or equal to about 380nm, greater than or equal to about 390nm, greater than or equal to about 400nm, greater than or equal to about 410nm, greater than or equal to about 420nm, greater than or equal to about 430nm, greater than or equal to about 440nm, greater than or equal to about 450nm, greater than or equal to about 500nm, greater than or equal to about 500nm, or any range or value between these values.

[0108] In some embodiments, the average diameter of the liposomes is any range or value between about 50 nm and about 500 nm, about 50 nm and about 400 nm, about 50 nm and about 300 nm, about 50 nm and about 250 nm, about 100 nm and about 500 nm, about 100 nm and about 400 nm, about 100 nm and about 300 nm, about 100 nm and about 250 nm, about 200 nm and about 500 nm, about 200 nm and about 400 nm, about 200 nm and about 300 nm. In some embodiments, the average diameter of the liposomes is about 250 nm.

[0109] In some embodiments, the polydispersity index (PdI) of the liposomes is from 0 to about 0.2. In some embodiments, the polydispersity index is about 0.01, 0.025, 0.05, 0.1, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8. In some instances, the polydispersity index is in the range of about 0.01 to about 0.8, about 0.025 to about 0.75, about 0.05 to about 0.6, or about 0.1 to about 0.3.

[0110] In some embodiments, the liposomes comprise propylene glycol, lecithin, or a combination thereof. In some embodiments, propylene glycol, by weight, is greater than or equal to about 0.001 wt.%, greater than or equal to about 0.005 wt.%, greater than or equal to about 0.01 wt.%, greater than or equal to about 0.02 wt.%, greater than or equal to about 0.05 wt.%, greater than or equal to about 0.10 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about 0.001 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about 0.50 ... The liposomes contain concentrations of about 2.0 wt.%, greater than or equal to about 2.5 wt.%, greater than or equal to about 3.0 wt.%, greater than or equal to about 3.5 wt.%, greater than or equal to about 4.0 wt.%, greater than or equal to about 4.5 wt.%, greater than or equal to about 5.0 wt.%, greater than or equal to about 5.5 wt.%, greater than or equal to about 6.0 wt.%, greater than or equal to about 6.5 wt.%, greater than or equal to about 7.0 wt.%, greater than or equal to about 8 wt.%, greater than or equal to about 9 wt.%, greater than or equal to about 10 wt.%, or any range or value between these values.

[0111] In some embodiments, propylene glycol is present in the liposomes at a concentration of about 0.001 wt.% to about 6 wt.%, about 0.002 wt.% to about 4 wt.%, about 0.01 wt.% to about 3 wt.%, or about 0.02 wt.% to about 2 wt.% (by weight) relative to the total weight of the liposomes or the total weight of the topical composition.

[0112] In some embodiments, lecithin, by weight, relative to the total weight of liposomes, is greater than or equal to about 0.001 wt.%, greater than or equal to about 0.005 wt.%, greater than or equal to about 0.01 wt.%, greater than or equal to about 0.02 wt.%, greater than or equal to about 0.05 wt.%, greater than or equal to about 0.10 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about 0.001 ...005 wt.%, greater than or equal to about 0.01 wt.%, greater than or equal to about 0.02 wt.%, greater than or equal to about 0.05 wt.%, greater than or equal to about 0.10 wt.%, greater than or equal to about 0.20 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.25 wt.%, greater than or equal to about 0.50 wt.%, greater than or equal to about 0.75 wt.%, greater than or equal to about 1.0 wt.%, greater than or equal to about 1.5 wt.%, greater than or equal to about The liposomes contain concentrations of about 2.0 wt.%, greater than or equal to about 2.5 wt.%, greater than or equal to about 3.0 wt.%, greater than or equal to about 3.5 wt.%, greater than or equal to about 4.0 wt.%, greater than or equal to about 4.5 wt.%, greater than or equal to about 5.0 wt.%, greater than or equal to about 5.5 wt.%, greater than or equal to about 6.0 wt.%, greater than or equal to about 6.5 wt.%, greater than or equal to about 7.0 wt.%, greater than or equal to about 8 wt.%, greater than or equal to about 9 wt.%, greater than or equal to about 10 wt.%, or any range or value between these values.

[0113] In some embodiments, lecithin is present in the liposomes at a concentration of about 0.001 wt.% to about 6 wt.%, about 0.002 wt.% to about 4 wt.%, about 0.01 wt.% to about 3 wt.%, or about 0.02 wt.% to about 2 wt.% (by weight) relative to the total weight of the liposomes or the total weight of the topical composition.

[0114] In some embodiments, liposomes are commercially available (e.g., PRO-LIPO). TM In some embodiments, the liposomes comprise propylene glycol, lecithin, sunflower seed oil, and tocopherol.

[0115] Alpha-hydroxy acid (AHA)

[0116] The local compositions according to this disclosure comprise one or more alpha-hydroxy acids (AHAs). In some embodiments, the one or more AHAs are selected from the group consisting of: mandelic acid, glycolic acid, lactic acid, citric acid, malic acid, hydroxyoctanoic acid, and hydroxydecanoic acid. In some embodiments, the one or more AHAs comprise mandelic acid. In some embodiments, mandelic acid comprises (S)-mandelic acid. In some embodiments, mandelic acid comprises (R)-mandelic acid. In some embodiments, mandelic acid comprises both (S)-mandelic acid and (R)-mandelic acid (e.g., para-mandelic acid).

[0117] Mandelic acid is derived from almonds and is a larger molecule than other AHAs used in skincare products (e.g., glycolic acid). Therefore, it tends to penetrate the skin more slowly, resulting in less skin irritation compared to other AHAs. At low concentrations (e.g., 6 wt.% or lower), said mandelic acid does not cause significant skin peeling, is safe for home use, and is tolerable for patients with various skin types. Therefore, mandelic acid is a viable option for topical application to improve skin quality. See, for example, SW Jacobs et al., “Effects of Topical Mandelic Acid Treatment on Facial Skin Viscoelasticity,” 34 Facial Plast. Surg. 651–56 (2018).

[0118] One or more AHAs (e.g., mandelic acid) are present in the topical composition at any suitable concentration suitable for achieving therapeutic benefits (e.g., reducing cellular senescence). In some embodiments, one or more AHAs (e.g., mandelic acid) are present in the topical composition at a concentration of at least about 0.01 wt.%, at least about 0.02 wt.%, at least about 0.03 wt.%, at least about 0.04 wt.%, at least about 0.05 wt.%, at least about 0.06 wt.%, at least about 0.07 wt.%, at least about 0.08 wt.%, at least about 0.09 wt.%, at least about 0.10 wt.%, at least about 0.15 wt.%, at least about 0.20 wt.%, at least about 0.25 wt.%, at least about 0.30 wt.%, at least about 0.35 wt.%, at least about 0.40 wt.%, at least about 0.45 wt.%, and less than 0.50 wt.%, or any range or value between thereof, relative to the total weight of the topical composition.

[0119] In some embodiments, one or more AHAs (e.g., mandelic acid) are present in the topical composition at concentrations of less than 0.50 wt.%, less than or equal to about 0.45 wt.%, less than or equal to about 0.40 wt.%, less than or equal to about 0.35 wt.%, less than or equal to about 0.30 wt.%, less than or equal to about 0.25 wt.%, less than or equal to about 0.20 wt.%, less than or equal to about 0.15 wt.%, less than or equal to about 0.10 wt.%, less than or equal to about 0.09 wt.%, less than or equal to about 0.08 wt.%, less than or equal to about 0.07 wt.%, less than or equal to about 0.06 wt.%, less than or equal to about 0.05 wt.%, or any range or value between these values ​​relative to the total weight of the topical composition.

[0120] In some embodiments, one or more AHAs (e.g., mandelic acid) are present in the topical composition at concentrations of at least about 0.01 wt.% and less than 0.5 wt.%, about 0.01 wt.% to about 0.4 wt.%, about 0.01 wt.% to about 0.3 wt.%, about 0.01 wt.% to about 0.2 wt.%, about 0.01 wt.% to about 0.1 wt.%, about 0.01 wt.% to about 0.05 wt.%, about 0.01 wt.% to about 0.04 wt.%, about 0.01 wt.% to about 0.03 wt.%, 0.05 wt.% to about 0.5 wt.%, about 0.05 wt.% to about 0.4 wt.%, about 0.05 wt.% to about 0.3 wt.%, about 0.05 wt.% to about 0.2 wt.%, about 0.05 wt.% to about 0.1 wt.%, or any range or value thereof, relative to the total weight of the topical composition.

[0121] In some embodiments, one or more AHAs (e.g., mandelic acid) are present in the topical composition at concentrations lower than the recommended concentrations for formulating one or more AHAs into a topical composition (e.g., the recommended concentration of mandelic acid for skin brightening of about 0.5 wt.% to 6 wt.%). The inventors have surprisingly discovered that low concentrations of AHAs, and particularly mandelic acid (e.g., below 0.5 wt.%), produce numerous therapeutic benefits, which are discussed below (see Method of Use). Simultaneously, low concentrations of AHAs achieve the desired acidic pH of the topical composition (e.g., 4 to 6, 3 to 5, or 4 to 5) while reducing or eliminating undesirable effects associated with use at higher doses, such as inflammation and skin irritation.

[0122] The inventors also surprisingly discovered that low doses of AHAs (e.g., mandelic acid) and low doses of Centella asiatica extract exhibit synergistic activity, such as synergistic protection against the effects of senescent cell byproducts and reduction of inflammation in the epidermis and increase of cell cycle activity in the dermis. This activity implies a synergistic effect in reducing cellular senescence, a synergy that was unexpected and unknown in the art at the time.

[0123] Centella asiatica extract and AHA are present in any weight ratio suitable for achieving the synergistic activities mentioned above. In some embodiments, Centella asiatica extract and AHA are present in a weight ratio of about 1:2 to 1:20, about 1:5 to 1:15, about 1:6 to 1:14, about 1:7 to about 1:13, about 1:8 to about 1:12, or any range or value between these weights. In some embodiments, Centella asiatica extract and AHA are present in a weight ratio of about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, or less.

[0124] In some embodiments, the triterpenoid compound is present with AHA in a weight ratio of about 1:2 to 1:400, about 1:5 to 1:200, about 1:6 to 1:100, about 1:7 to about 1:50, about 1:8 to about 1:20, about 1:10 to 1:15 or any range or value therebetween. In some embodiments, the triterpenoid compound is present with AHA in a weight ratio of about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:25, about 1:30, about 1:35, about 1:40, about 1:45, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:150, about 1:200, about 1:300, about 1:400 or less.

[0125] antioxidants

[0126] UV irradiation induces oxidative stress, which is detrimental to cellular function and can negatively impact cell survival. Antioxidants can mitigate these oxidative processes by counteracting the formation or spread of reactive oxygen species, thereby improving cellular function and survival.

[0127] In some embodiments, the topical composition according to this disclosure comprises one or more antioxidants. In some embodiments, the one or more antioxidants comprise tocopherol, tocopheryl acetate, hydroxyacetophenone, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tert-butylhydroquinone (TBHQ), propyl gallate, tocotrienol, ascorbyl palmitate, rosemary (Rosmarinus officinalis / rosemary) leaf extract, or combinations thereof. In some embodiments, the one or more antioxidants comprise tocopherol, tocopheryl acetate, or combinations thereof. In some embodiments, the one or more antioxidants comprise tocopherol. In some embodiments, the one or more antioxidants comprise tocopheryl acetate. In some embodiments, the one or more antioxidants are present separately from any antioxidants in the liposomes or composition that may be present in the liposomes. In some embodiments, the one or more antioxidants are present in an oil phase prepared separately from the liposomes or composition.

[0128] In some embodiments, one or more antioxidants may be present, alone or in combination, at least about 0.001 wt.%, at least about 0.002 wt.%, at least about 0.003 wt.%, at least about 0.004 wt.%, at least about 0.005 wt.%, at least about 0.006 wt.%, at least about 0.007 wt.%, at least about 0.008 wt.%, at least about 0.009 wt.%, at least about 0.01 wt.%, at least about 0.02 wt.%, at least about 0.03 wt.%, at least about 0.04 wt.%, at least about 0.05 wt.%, at least about 0.06 wt.%, at least about 0.07 wt.%, at least about 0.08 wt.%, at least about 0.09 wt.%, at least about 0.1 wt.%, at least about 0.2 wt.%, at least about 0.3 wt.%, at least about 0.4 wt.% of the total weight of the composition. .%, at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, at least about 2.0 wt.%, at least about 2.5 wt.%, at least about 3.0 wt.%, at least about 3.5 wt.%, at least about 4.0 wt.% The concentration is present at a range or value of at least 4.5 wt.%, at least 5.0 wt.%, at least 5.5 wt.%, at least 6.0 wt.%, at least 6.5 wt.%, at least 7.0 wt.%, at least 7.5 wt.%, at least 8.0 wt.%, at least 8.5 wt.%, at least 9.0 wt.%, at least 9.5 wt.%, at least 10 wt.%, or between these ranges or values. In some embodiments, one or more antioxidants are present at these concentrations separately from any antioxidants in the liposomes or composition that may be present in the liposomes or composition (e.g., in addition to these antioxidants).

[0129] In some embodiments, one or more antioxidants may be present, alone or in combination, at a weight percentage relative to the total weight of the composition of no more than about 10.0 wt.%, no more than about 9.5 wt.%, no more than about 9.0 wt.%, no more than about 8.5 wt.%, no more than about 8.0 wt.%, no more than about 7.5 wt.%, no more than about 7.0 wt.%, no more than about 6.5 wt.%, no more than about 6.0 wt.%, no more than about 5.5 wt.%, no more than about 5.0 wt.%, no more than about 4.5 wt.%, no more than about 4.0 wt.%, no more than about 3.5 wt.%, no more than about 3.0 wt.%, no more than about 2.5 wt.%, no more than about 2.0 wt.%, and no more than about 1.5 wt.%. The concentration is present in any range or value between approximately 1.0 wt.%, approximately 0.9 wt.%, approximately 0.8 wt.%, approximately 0.7 wt.%, approximately 0.6 wt.%, approximately 0.5 wt.%, approximately 0.4 wt.%, approximately 0.3 wt.%, approximately 0.2 wt.%, approximately 0.1 wt.%, approximately 0.09 wt.%, approximately 0.08 wt.%, approximately 0.07 wt.%, approximately 0.06 wt.%, approximately 0.05 wt.%, approximately 0.04 wt.%, approximately 0.03 wt.%, approximately 0.02 wt.%, approximately 0.01 wt.%, or within the range of such concentrations. In some embodiments, one or more antioxidants are present at these concentrations separately from any antioxidants in liposomes that may be present in the liposomes or the composition (e.g., in addition to these antioxidants).

[0130] In some embodiments, one or more antioxidants may be present, alone or in combination, in amounts of about 0.001 wt.%, about 0.002 wt.%, about 0.005 wt.%, about 0.008 wt.%, about 0.01 wt.%, about 0.02 wt.%, about 0.05 wt.%, about 0.08 wt.%, about 0.1 wt.%, about 0.2 wt.%, about 0.5 wt.%, about 0.8 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, or about 2.2 wt.% of the total weight of the composition, relative to the total weight of the composition. The concentrations are present at concentrations of approximately 10.0%, about 2.5 wt.%, about 2.8 wt.%, about 3.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, about 4.0 wt.%, about 4.2 wt.%, about 4.5 wt.%, about 4.8 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 6.0 wt.%, about 6.5 wt.%, about 7.0 wt.%, about 7.5 wt.%, about 8.0 wt.%, about 8.5 wt.%, about 9.0 wt.%, about 9.5 wt.%, or any range or value between these concentrations. In some embodiments, one or more antioxidants are present at these concentrations separately from any antioxidants in the delivery medium (e.g., liposomes) that may be present in the composition (e.g., in addition to the said antioxidants).

[0131] In some embodiments, one or more antioxidants, alone or together, are present in the composition at concentrations ranging from about 0.001 wt.% to about 10 wt.%, about 0.01 wt.% to about 10 wt.%, about 0.1 wt.% to about 10 wt.%, about 1 wt.% to about 10 wt.%, about 0.001 wt.% to about 1 wt.%, about 0.01 wt.% to about 1 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.001 wt.% to about 0.1 wt.%, about 0.01 wt.% to about 10 wt.%, about 0.01 wt.% to about 1 wt.%, about 0.01 wt.% to about 0.1 wt.%, about 0.1 wt.% to about 1 wt.%, or between any range or value thereof, relative to the total weight of the composition. In some embodiments, one or more antioxidants are present at these concentrations separately from any antioxidants in the delivery medium (e.g., in addition to the antioxidants) that may be present in the composition.

[0132] Moisturizer / Emollient

[0133] In some embodiments, the topical composition according to this disclosure comprises one or more moisturizers and / or emollients. By way of non-limiting examples, in some embodiments, one or more moisturizers and / or emollients may comprise polyols, such as polyols having 2 to 20 carbon atoms, including glycerol (glycerol / glycerin); ethylene glycol derivatives (e.g., propylene glycol, butylene glycol, pentylene glycol, hexanediol, dipropylene glycol, diethylene glycol, octyl ethylene glycol); and mixtures thereof. In some embodiments, the moisturizer and / or emollient may comprise glyceryl sorbitol; sugars (e.g., glucose, lactose, etc.); alkoxylated glucose derivatives; glucose ethers; panthenol (e.g., D-panthenol, DL-panthenol); polyethylene glycol (PEG); urea; sodium hyaluronate; octyl glycol; panthenol lactone; caprylic / capric triglyceride; cocoyl oleate / capric oleate; panthenol lactone; squalane; soluble chitosan; glycerol; amino acids (e.g., serine, citrulline, arginine, asparagine, or alanine); alpha-hydroxy acids; salicylic acid; hyaluronic acid (HA); sodium lactate; sodium pyroglutamic acid / sodium PCA; aloe vera gel; or combinations thereof.

[0134] In some embodiments, one or more moisturizers and / or emollients may be used alone or in combination in amounts of at least about 0.1 wt.%, at least about 0.2 wt.%, at least about 0.3 wt.%, at least about 0.4 wt.%, at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, or at least about 2.0 wt.% relative to the total weight of the composition. The concentration is present in at least about 2.5 wt.%, at least about 3.0 wt.%, at least about 3.5 wt.%, at least about 4.0 wt.%, at least about 4.5 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, at least about 10 wt.%, or any range or value between these values.

[0135] In some embodiments, one or more moisturizers and / or emollients may be present alone or in combination in amounts not exceeding about 30 wt.%, not exceeding about 25 wt.%, not exceeding about 20 wt.%, not exceeding about 15 wt.%, not exceeding about 14 wt.%, not exceeding about 13 wt.%, not exceeding about 12 wt.%, not exceeding about 11 wt.%, not exceeding about 10.0 wt.%, not exceeding about 9.5 wt.%, and not exceeding about 9.0 wt.% relative to the total weight of the composition. The concentration is present in any range or value between the following concentrations: % and not greater than about 8.5 wt.%, not greater than about 8.0 wt.%, not greater than about 7.5 wt.%, not greater than about 7.0 wt.%, not greater than about 6.5 wt.%, not greater than about 6.0 wt.%, not greater than about 5.5 wt.%, not greater than about 5.0 wt.%, not greater than about 4.5 wt.%, not greater than about 4.0 wt.%, not greater than about 3.5 wt.%, not greater than about 3.0 wt.%.

[0136] In some embodiments, one or more moisturizers and / or emollients may be used alone or in combination in amounts of about 0.1 wt.%, about 0.2 wt.%, about 0.5 wt.%, about 0.8 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, about 2.2 wt.%, about 2.5 wt.%, about 2.8 wt.%, about 3.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, relative to the total weight of the composition. The concentration is present in approximately 4.0 wt.%, approximately 4.2 wt.%, approximately 4.5 wt.%, approximately 4.8 wt.%, approximately 5.0 wt.%, approximately 5.5 wt.%, approximately 6.0 wt.%, approximately 6.5 wt.%, approximately 7.0 wt.%, approximately 7.5 wt.%, approximately 8.0 wt.%, approximately 8.5 wt.%, approximately 9.0 wt.%, approximately 9.5 wt.%, approximately 10.0 wt.%, approximately 15 wt.%, approximately 20 wt.%, approximately 25 wt.%, approximately 30 wt.%, or any range or value between these values.

[0137] In some embodiments, one or more moisturizers and / or emollients may be used alone or together in amounts relative to the total weight of the composition, such as from about 0.1 wt.% to about 30 wt.%, from about 0.1 wt.% to about 25 wt.%, from about 0.1 wt.% to about 20 wt.%, from about 0.1 wt.% to about 15 wt.%, from about 0.1 wt.% to about 14 wt.%, from about 0.1 wt.% to about 13 wt.%, from about 0.1 wt.% to about 12 wt.%, from about 0.1 wt.% to about 11 wt.%, from about 0.1 wt.% to about 10.0 wt.%, or from about 0.1 wt.% About 8 wt.%, about 0.1 wt.% to about 5 wt.%, about 0.1 to about 4 wt.%, about 0.1 wt.% to about 3 wt.%, about 0.1 wt.% to about 2 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.5 wt.% to about 30 wt.%, about 0.5 wt.% to about 25 wt.%, about 0.5 wt.% to about 20 wt.%, about 0.5 wt.% to about 15 wt.%, about 0.5 wt.% to about 10.0 wt.%, about 0.5 wt.% to about 8 wt.%, about 0.5 wt.% to about 5 wt.%, about 0.5 to about 4 wt.%, about 0.5 wt.% to about 3 wt.%, about 0.5 wt.% to about 2 wt.%, about 0.5 to about 1 wt.%, about 1 wt.% to about 30 wt.%, about 1 wt.% to about 25 wt.%, about 1 wt.% to about 20 wt.%, about 1 wt.% to about 15 wt.%, about 1 wt.% to about 10 wt.%, about 1 wt.% to about 8 wt.%, about 1 wt.% to about 5 wt.%, about 1 wt.% to about 4 wt.%, about 1 wt.% to about 3 wt.%, about 1 wt.% to about 2 wt.%, about 3 wt.% The concentration is present in any range or value between t.% and 30 wt.%, about 3 wt.% and 25 wt.%, about 3 wt.% and 20 wt.%, about 3 wt.% and 15 wt.%, about 1 wt.% and 10 wt.%, about 3 wt.% and 8 wt.%, about 3 wt.% and 5 wt.%, about 5 wt.% and 30 wt.%, about 5 wt.% and 25 wt.%, about 5 wt.% and 20 wt.%, about 5 wt.% and 15 wt.%, about 5 wt.% and 10 wt.%, about 5 wt.% and 8 wt.%, or between.

[0138] emulsifier

[0139] The topical compositions according to this disclosure may contain one or more emulsifiers for stabilizing the aqueous and / or oil phases of the emulsion. Suitable emulsifiers for topical compositions include anionic surfactants, cationic surfactants, nonionic surfactants, and amphoteric surfactants.

[0140] In some embodiments, the emulsifier comprising polyol esters can be used as emulsifiers or emollients. Suitable polyol esters include ethylene glycol mono- and di-fatty acid esters, diethylene glycol mono- and di-fatty acid esters, polyethylene glycol (200-6000) mono- and di-fatty acid esters, propylene glycol mono- and di-fatty acid esters, polypropylene glycol 2000 monooleate, polypropylene glycol 2000 monostearate, ethoxylated propylene glycol monostearate, glycerol mono- and di-fatty acid esters, polyglycerol poly-fatty acid esters, ethoxylated glycerol monostearate, 1,3-butanediol monostearate, 1,3-butanediol distearate, polyoxyethylene polyol fatty acid esters, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene ethers of highly saturated fatty alcohols, and alkyl polyglycoside emulsifiers. In some embodiments, the emulsifier comprises acrylic polymers (e.g., acrylate / C10-30 alkyl acrylate crosspolymers, such as those named PEMULEN). TM (EZ-4U is a crosslinked polymer). In some embodiments, the emulsifier comprises potassium cetyl phosphate. In some embodiments, the emulsifier comprises methylglucose sesquistearate.

[0141] In some embodiments, the emulsifier may be used alone or in combination at a concentration of at least about 0.1 wt.%, at least about 0.2 wt.%, at least about 0.3 wt.%, at least about 0.4 wt.%, at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, at least about 2.0 wt.%, at least about 2.2 wt.%, at least about 2.5 wt.%, at least about 2.8 wt.%, or at least about 3 wt.% of the total weight of the composition. The concentration is present at 0 wt.%, at least about 3.2 wt.%, at least about 3.5 wt.%, at least about 3.8 wt.%, at least about 4.0 wt.%, at least about 4.2 wt.%, at least about 4.5 wt.%, at least about 4.8 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, at least about 10.0 wt.%, or any range or value between these values.

[0142] In some embodiments, the emulsifier may be present alone or in combination at a concentration of no more than about 10.0 wt.%, no more than about 9.5 wt.%, no more than about 9.0 wt.%, no more than about 8.5 wt.%, no more than about 8.0 wt.%, no more than about 7.5 wt.%, no more than about 7.0 wt.%, no more than about 6.5 wt.%, no more than about 6.0 wt.%, no more than about 5.5 wt.%, no more than about 5.0 wt.%, no more than about 4.5 wt.%, no more than about 4.0 wt.%, no more than about 3.5 wt.%, no more than about 3.0 wt.%, or any range or value between thereof, relative to the total weight of the composition.

[0143] In some embodiments, the emulsifier may be used alone or in combination at a concentration relative to the total weight of the composition of about 0.1 wt.%, about 0.2 wt.%, about 0.3 wt.%, about 0.4 wt.%, about 0.5 wt.%, about 0.6 wt.%, about 0.7 wt.%, about 0.8 wt.%, about 0.9 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, about 2.2 wt.%, about 2.5 wt.%, about 2.8 wt.%, about 3 wt.%, or about 3 wt.%. The concentration is present in any range or value between 0.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, about 4.0 wt.%, about 4.2 wt.%, about 4.5 wt.%, about 4.8 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 6.0 wt.%, about 6.5 wt.%, about 7.0 wt.%, about 7.5 wt.%, about 8.0 wt.%, about 8.5 wt.%, about 9.0 wt.%, about 9.5 wt.%, about 10.0 wt.%, or between.

[0144] In some embodiments, the emulsifier may be used alone or in combination in amounts relative to the total weight of the composition, such as from about 0.1 wt.% to about 10 wt.%, from about 0.1 wt.% to about 5 wt.%, from about 0.1 wt.% to about 4 wt.%, from about 0.1 wt.% to about 3 wt.%, from about 0.1 wt.% to about 2 wt.%, from about 0.1 wt.% to about 1 wt.%, from about 0.1 wt.% to about 0.5 wt.%, from about 0.5 wt.% to about 10 wt.%, from about 0.5 wt.% to about 5 wt.%, from about 0.5 wt.% to about 4 wt.%, and from about 0.5 wt.%. The concentration is present in any range or value between % and 3 wt.%, about 0.5 wt.% and about 2 wt.%, about 0.5 wt.% and about 1 wt.%, about 1 wt.% and about 10 wt.%, about 1 wt.% and about 5 wt.%, about 1 wt.% and about 4 wt.%, about 1 wt.% and about 3 wt.%, about 1 wt.% and about 2 wt.%, about 2 wt.% and about 10 wt.%, about 2 wt.% and about 5 wt.%, about 3 wt.% and about 10 wt.%, about 3 wt.% and about 5 wt.%, about 5 wt.% and about 10 wt.%, or between.

[0145] Oil

[0146] In some embodiments, the local composition according to this disclosure comprises one or more oils. In some embodiments (e.g., when the composition is in emulsion form, such as an oil-in-water emulsion or an oil-in-water emulsion), the composition comprises an oil phase comprising one or more oils. By way of non-limiting examples, the one or more oils may comprise vegetable oils, mineral oils, animal oils or synthetic waxes, synthetic oils or synthetic butter, and mixtures thereof. In some embodiments, the oil may comprise: one or more mineral oils (e.g., those sold by Esso). and One or more vegetable oils (e.g., almond oil, sweet almond oil, palm oil, soybean oil, sesame oil, sunflower oil, olive oil, etc.); hydrogenated vegetable oils; one or more animal oils or plant-derived substitutes (e.g., lanolin, squalene, or fish oil and their derivatives, such as perhydrosqualene, for example, Sophim). (sales); one or more synthetic oils (e.g., cetearyl isononanoate, such as Cognis France's CETIOL). Isonononyl isononanoate, such as DUB sold by Stéarinerie Dubois. Diisopropyl adipic acid, such as Croda's CRODAMOL. Isopropyl palmitate, such as Croda's CRODAMOL Caprylic / capric triglycerides, such as those sold by Univar. Hydrogenated polyisobutylene, such as that from Rossow Products); one or more silicone oils (e.g., dimethicone, such as Dow Corning's Q7-9120 with a viscosity of 20 cSt and an ester of 12,500 cSt). Cyclomethyl silicone oils, such as Dow Corning's ST-cyclodimethylsiloxane Or Dow Corning's DC 9045 ); or any combination thereof.

[0147] In some embodiments, one or more oils are present alone or together in amounts of at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, at least about 2.0 wt.%, at least about 2.5 wt.%, at least about 3.0 wt.%, at least about 3.5 wt.%, at least about 4 wt.% of the total weight of the composition. The concentration is present in any range or value between 0.0 wt.%, at least about 4.5 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, at least about 10 wt.%, at least about 15.0 wt.%, at least about 20.0 wt.%, or between.

[0148] In some embodiments, one or more oils may be used alone or in combination in amounts not exceeding about 35.0 wt.%, not exceeding about 30 wt.%, not exceeding about 25 wt.%, not exceeding about 20 wt.%, not exceeding about 19 wt.%, not exceeding about 18 wt.%, not exceeding about 17 wt.%, not exceeding about 16 wt.%, not exceeding about 15 wt.%, not exceeding about 14 wt.%, not exceeding about 13 wt.%, not exceeding about 12 wt.%, not exceeding about 11 wt.%, and not exceeding about 10.0 wt.% of the total weight of the composition. The concentration is present in any range or value between 0.0% and approximately 9.5 wt.%, 9.0 wt.%, 8.5 wt.%, 8.0 wt.%, 7.5 wt.%, 7.0 wt.%, 6.5 wt.%, 6.0 wt.%, 5.5 wt.%, 5.0 wt.%, 4.5 wt.%, 4.0 wt.%, 3.5 wt.%, or 3.0 wt.%.

[0149] In some embodiments, one or more oils may be used alone or together in amounts of about 0.5 wt.%, about 0.8 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, about 2.2 wt.%, about 2.5 wt.%, about 2.8 wt.%, about 3.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, about 4.0 wt.%, about 4.2 wt.%, about 4.5 wt.%, about 4.8 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 6.0 wt.%, about 6.5 wt.%, about 7.0 wt.%, about 7.5 wt.%, about 8.0 wt.%, about 8.5 wt.%, about 9 wt.% relative to the total weight of the composition. The concentration is present in any range or value between 0 wt.%, about 9.5 wt.%, about 10.0 wt.%, about 10.5 wt.%, about 11.0 wt.%, about 11.5 wt.%, about 12.0 wt.%, about 12.5 wt.%, about 13.0 wt.%, about 13.5 wt.%, about 14.0 wt.%, about 14.5 wt.%, about 15.0 wt.%, about 16 wt.%, about 17 wt.%, about 18 wt.%, about 19 wt.%, about 20 wt.%, about 21 wt.%, about 22 wt.%, about 23 wt.%, about 24 wt.%, about 25 wt.%, about 26 wt.%, about 27 wt.%, about 28 wt.%, about 29 wt.%, about 30 wt.%, about 35 wt.%, or between these values.

[0150] In some embodiments, one or more oils may be used alone or together in amounts relative to the total weight of the composition, such as about 0.5 wt.% to about 35 wt.%, about 0.5 wt.% to about 30 wt.%, about 0.5 wt.% to about 25 wt.%, about 0.5 wt.% to about 20 wt.%, about 0.5 wt.% to about 15 wt.%, about 0.5 wt.% to about 10.0 wt.%, about 0.5 wt.% to about 5 wt.%, about 0.5 wt.% to about 4 wt.%, about 0.5 wt.% to about 3 wt.%, about 0.5 wt.% to about 2 wt.%, about 0.5 wt.% to about 1 wt.%, about 1 wt.% to about 30 wt.%, about 1 wt.% to about 25 wt.%. The concentration is present in any range or value between about 1 wt.% and about 20 wt.%, about 1 wt.% and about 15 wt.%, about 1 wt.% and about 10 wt.%, about 1 wt.% and about 8 wt.%, about 1 wt.% and about 5 wt.%, about 1 wt.% and about 4 wt.%, about 1 wt.% and about 3 wt.%, about 1 wt.% and about 2 wt.%, about 3 wt.% and about 30 wt.%, about 3 wt.% and about 20 wt.%, about 3 wt.% and about 15 wt.%, about 3 wt.% and about 10 wt.%, about 3 wt.% and about 8 wt.%, about 3 wt.% and about 5 wt.% and about 10 wt.%, about 5 wt.% and about 8 wt.%, or between.

[0151] Other fatty phase components

[0152] The compositions according to this disclosure may comprise an oil phase or a fatty phase. In some embodiments, the oil phase or fatty phase comprises oil and other components (e.g., waxes, fatty alcohols, fatty acids, esters of carboxylic acids or diacids, alkyl esters of fatty acids or diacids, alkenyl esters of fatty acids or diacids, mineral oils, petrolatum, etc.).

[0153] In some embodiments, the partial compositions according to this disclosure comprise one or more waxes. In some embodiments, the wax is selected from lanolin and its derivatives, including lanolin oil, lanolin wax, lanolin alcohol, lanolin fatty acid, isopropyl lanolinate, ethoxylated lanolin, ethoxylated lanolin alcohol, ethoxylated cholesterol, propoxylated lanolin alcohol, acetylated lanolin, acetylated lanolin alcohol, lanolin alcohol linoleate, lanolin alcohol ricinoleate, lanolin alcohol ricinoleate acetate, lanolin alcohol ricinoleate acetate, ethoxylated alcohol ester acetate, hydrogenated lanolin, hydrogenated lanolin, ethoxylated hydrogenated lanolin, ethoxylated sorbitol lanolin, and liquid and semi-solid lanolin. Materials that can also be used as waxes include hydrocarbon waxes, ester waxes, amide waxes, microcrystalline waxes, hydrogenated vegetable oils, rice bran wax, sunflower seed wax, paraffin wax, ceresin, and combinations thereof. Materials that can be used as waxes are wax esters, such as beeswax (e.g., synthetic beeswax), cetearyl, myristate, and stearate; beeswax derivatives, such as polyoxyethylene sorbitol beeswax; and plant waxes, including carnauba wax and candelilla wax.

[0154] In some embodiments, the partial compositions according to this disclosure comprise one or more fatty acids (e.g., nonanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, hydroxystearic acid, oleic acid, linoleic acid, ricinoleic acid, arachidic acid, behenic acid, erucic acid, and combinations thereof). In some embodiments, the partial compositions according to this disclosure comprise one or more fatty alcohols (e.g., lauryl alcohol, myristic alcohol, cetyl alcohol, hexadecyl alcohol, stearyl alcohol, isostearyl alcohol, hydroxystearyl alcohol, oleyl alcohol, ricinoleyl alcohol, behenyl alcohol, and arugula alcohol, and combinations thereof, such as octadecyl alcohol (i.e., cetearyl alcohol)) and 2-octyldodecyl alcohol.

[0155] In some embodiments, the topical compositions according to this disclosure comprise esters of carboxylic acids or dicarboxylic acids (e.g., methyl, isopropyl, and butyl esters of fatty acids). In some embodiments, the topical compositions according to this disclosure comprise alkyl esters (e.g., hexyl laurate, isohexyl laurate, isohexyl palmitate, isopropyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, isopropyl isostearate, dilauryl lactate, myristyl lactate, and hexadecyl lactate). In some embodiments, the topical compositions according to this disclosure comprise alkenyl esters of fatty acids, such as oleyl myristate, oleyl stearate, and oleyl oleate. In some embodiments, the topical compositions according to this disclosure comprise alkyl esters of dicarboxylic acids (e.g., diisopropyl adipate, diisohexyl adipate, bis(hexyldecyl) adipate, and diisopropyl sebacate).

[0156] In some embodiments, additional aliphatic phase components (e.g., waxes, fatty alcohols, fatty acids, esters of carboxylic acids or diacids, alkyl esters of fatty acids or diacids, alkenyl esters of fatty acids or diacids) may be present, alone or together, in amounts of at least about 0.1 wt.%, at least about 0.2 wt.%, at least about 0.3 wt.%, at least about 0.4 wt.%, at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, at least about 2.0 wt.%, at least about 2.2 wt.%, to [amount missing] wt.% of the total weight of the composition. The concentration is present in any range or value between approximately 2.5 wt.%, at least approximately 2.8 wt.%, at least approximately 3.0 wt.%, at least approximately 3.2 wt.%, at least approximately 3.5 wt.%, at least approximately 3.8 wt.%, at least approximately 4.0 wt.%, at least approximately 4.2 wt.%, at least approximately 4.5 wt.%, at least approximately 4.8 wt.%, at least approximately 5.0 wt.%, at least approximately 5.5 wt.%, at least approximately 6.0 wt.%, at least approximately 6.5 wt.%, at least approximately 7.0 wt.%, at least approximately 7.5 wt.%, at least approximately 8.0 wt.%, at least approximately 8.5 wt.%, at least approximately 9.0 wt.%, at least approximately 9.5 wt.%, at least approximately 10.0 wt.%, or between these values.

[0157] In some embodiments, additional aliphatic phase components (e.g., waxes, fatty alcohols, fatty acids, esters of carboxylic acids or diacids, alkyl esters of fatty acids or diacids, alkenyl esters of fatty acids or diacids) may be present, alone or together, at concentrations not exceeding about 10.0 wt.%, not exceeding about 9.5 wt.%, not exceeding about 9.0 wt.%, not exceeding about 8.5 wt.%, not exceeding about 8.0 wt.%, not exceeding about 7.5 wt.%, not exceeding about 7.0 wt.%, not exceeding about 6.5 wt.%, not exceeding about 6.0 wt.%, not exceeding about 5.5 wt.%, not exceeding about 5.0 wt.%, not exceeding about 4.5 wt.%, not exceeding about 4.0 wt.%, not exceeding about 3.5 wt.%, not exceeding about 3.0 wt.%, or any range or value between thereof, relative to the total weight of the composition.

[0158] In some embodiments, additional aliphatic phase components (e.g., waxes, fatty alcohols, fatty acids, esters of carboxylic acids or diacids, alkyl esters of fatty acids or diacids, alkenyl esters of fatty acids or diacids) may be present alone or together in amounts of about 0.1 wt.%, about 0.2 wt.%, about 0.3 wt.%, about 0.4 wt.%, about 0.5 wt.%, about 0.6 wt.%, about 0.7 wt.%, about 0.8 wt.%, about 0.9 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, about 2.5 wt.%, or about 2.5 wt.%, relative to the total weight of the composition. The concentration is present in any range or value between 2 wt.%, about 2.5 wt.%, about 2.8 wt.%, about 3.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, about 4.0 wt.%, about 4.2 wt.%, about 4.5 wt.%, about 4.8 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 6.0 wt.%, about 6.5 wt.%, about 7.0 wt.%, about 7.5 wt.%, about 8.0 wt.%, about 8.5 wt.%, about 9.0 wt.%, about 9.5 wt.%, about 10.0 wt.%, or between.

[0159] In some embodiments, additional aliphatic phase components (e.g., waxes, fatty alcohols, fatty acids, esters of carboxylic acids or diacids, alkyl esters of fatty acids or diacids, alkenyl esters of fatty acids or diacids) may be present alone or together in amounts relative to the total weight of the composition of about 0.1 wt.% to about 10 wt.%, about 0.1 wt.% to about 5 wt.%, about 0.1 wt.% to about 4 wt.%, about 0.1 wt.% to about 3 wt.%, about 0.1 wt.% to about 2 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.1 wt.% to about 0.5 wt.%, about 0.5 wt.% to about 10 wt.%, about 0.5 wt.% to about 5 wt.% The concentration is present in any range or value between 0.5 wt.% and 4 wt.%, about 0.5 wt.% and 3 wt.%, about 0.5 wt.% and 2 wt.%, about 0.5 wt.% and 1 wt.%, about 1 wt.% and 10 wt.%, about 1 wt.% and 5 wt.%, about 1 wt.% and 4 wt.%, about 1 wt.% and 3 wt.%, about 1 wt.% and 2 wt.%, about 2 wt.% and 10 wt.%, about 2 wt.% and 5 wt.%, about 3 wt.% and 10 wt.%, about 3 wt.% and 5 wt.%, about 5 wt.% and 10 wt.%, or between.

[0160] gelling agent

[0161] In some embodiments, the topical composition according to this disclosure comprises one or more gelling agents (also referred to as suspending agents or thickeners). By way of non-limiting example, one or more gelling agents may comprise ready-to-use mixtures (e.g., those marketed by Sepico Corporation under the name...). Polyacrylate-13 / polyisobutylene / polysorbate 20 sold by Cypic Company under the name Ammonium acrylate / acrylamide copolymer / polyisobutylene / polysorbate 20 blend sold, or by Sepico Corporation under the name SIMULGEL TM Acrylamide / sodium acryloyldimethyl taurate copolymer / isohexadecane / polysorbate 80 sold in 600; acrylic polymers (e.g., acrylate / C10-30 alkyl acrylate crosspolymers, such as those under the name PEMULEN) TM Products sold by EZ-4U); carbomer (e.g., products sold by Lubrizol). CARBOPOL Or AQUA ); polysaccharides (e.g., xanthan gum, such as that sold by Kelco). Gellan gum (e.g., Kelco's) ), sclerotium gum (e.g., Alban Muller Industrie) ), guar gum and its derivatives (such as those from Rhodia under the name JAGUAR HP- The products sold include hydroxypropyl guar gum, cationic guar gum, pullullan, cellulose and cellulose derivatives (such as microcrystalline cellulose and sodium carboxymethyl cellulose, for example, Hercules under the name BLANOSE CMC). Sales of hydroxypropyl methylcellulose, such as those by Dow Chemical under the name METHOCEL Premium sales, hydroxyethyl cellulose, such as those from Aqualon under the name... Sales of methylcellulose, carboxymethylcellulose); magnesium aluminum silicate (e.g., sold by Vanderbilt University). VEEGUM or VEEGUM ); bentonite; modified starch (e.g., under the name STRUCTURE) Modified potato starch for sale); carrageenan (e.g., κ, λ, β, and ω family, such as those sold by IMCD). and Products); Polyvinyl alcohol (PVA) (e.g., Merck sells polyvinyl alcohol 40- Polyvinylpyrrolidone; carboxyvinyl polymers; acrylic / ethyl acrylate copolymers (e.g., Polyacrylic acid polymers; polymethacrylic acid polymers; polyvinyl acetate polymers; polyvinyl chloride polymers; polyvinylidene chloride polymers; mixtures of polyethylene glycol with polyethylene glycol stearate or distearate; oleogels (e.g., trihydroxystearin or aluminum magnesium stearate); nonionic polymers; and any combination of the above substances.

[0162] In some embodiments, one or more gelling agents may be used alone or in combination in amounts of at least about 0.1 wt.%, at least about 0.2 wt.%, at least about 0.3 wt.%, at least about 0.4 wt.%, at least about 0.5 wt.%, at least about 0.6 wt.%, at least about 0.7 wt.%, at least about 0.8 wt.%, at least about 0.9 wt.%, at least about 1.0 wt.%, at least about 1.2 wt.%, at least about 1.5 wt.%, at least about 1.8 wt.%, at least about 2.0 wt.%, at least about 2.2 wt.%, at least about 2.5 wt.%, at least about 2.8 wt.%, or at least about 0.5 wt.% of the total weight of the composition. The concentration is present in any range or value between 3.0 wt.%, at least about 3.2 wt.%, at least about 3.5 wt.%, at least about 3.8 wt.%, at least about 4.0 wt.%, at least about 4.2 wt.%, at least about 4.5 wt.%, at least about 4.8 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, at least about 10.0 wt.%, or between these values.

[0163] In some embodiments, one or more gelling agents may be present, alone or together, at concentrations not greater than about 10.0 wt.%, not greater than about 9.5 wt.%, not greater than about 9.0 wt.%, not greater than about 8.5 wt.%, not greater than about 8.0 wt.%, not greater than about 7.5 wt.%, not greater than about 7.0 wt.%, not greater than about 6.5 wt.%, not greater than about 6.0 wt.%, not greater than about 5.5 wt.%, not greater than about 5.0 wt.%, not greater than about 4.5 wt.%, not greater than about 4.0 wt.%, not greater than about 3.5 wt.%, not greater than about 3.0 wt.%, or any range or value between thereof, relative to the total weight of the composition.

[0164] In some embodiments, one or more gelling agents may be used alone or together in amounts relative to the total weight of the composition of about 0.1 wt.%, about 0.2 wt.%, about 0.3 wt.%, about 0.4 wt.%, about 0.5 wt.%, about 0.6 wt.%, about 0.7 wt.%, about 0.8 wt.%, about 0.9 wt.%, about 1.0 wt.%, about 1.2 wt.%, about 1.5 wt.%, about 1.8 wt.%, about 2.0 wt.%, about 2.2 wt.%, about 2.5 wt.%, about 2.8 wt.%, etc. The concentration is present in any range or value between about 3.0 wt.%, about 3.2 wt.%, about 3.5 wt.%, about 3.8 wt.%, about 4.0 wt.%, about 4.2 wt.%, about 4.5 wt.%, about 4.8 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 6.0 wt.%, about 6.5 wt.%, about 7.0 wt.%, about 7.5 wt.%, about 8.0 wt.%, about 8.5 wt.%, about 9.0 wt.%, about 9.5 wt.%, about 10.0 wt.%, or between.

[0165] In some embodiments, one or more gelling agents may be used alone or together in amounts relative to the total weight of the composition, such as about 0.1 wt.% to about 10 wt.%, about 0.1 wt.% to about 5 wt.%, about 0.1 wt.% to about 4 wt.%, about 0.1 wt.% to about 3 wt.%, about 0.1 wt.% to about 2 wt.%, about 0.1 wt.% to about 1 wt.%, about 0.1 wt.% to about 0.5 wt.%, about 0.5 wt.% to about 10 wt.%, about 0.5 wt.% to about 5 wt.%, about 0.5 wt.% to about 4 ... The concentration is present in any range or value between t.% and 3wt.%, about 0.5wt.% and 2wt.%, about 0.5wt.% and 1wt.%, about 1wt.% and 10wt.%, about 1wt.% and 5wt.%, about 1wt.% and 4wt.%, about 1wt.% and 3wt.%, about 1wt.% and 2wt.%, about 2wt.% and 10wt.%, about 2wt.% and 5wt.%, about 3wt.% and 10wt.%, about 3wt.% and 5wt.%, about 5wt.% and 10wt.%, or between.

[0166] composition pH

[0167] The topical compositions according to this disclosure may have any pH suitable for ensuring the chemical and physical stability of the composition and any active ingredients contained therein, and are non-irritating to the skin and prevent dry skin.

[0168] In some embodiments, the pH of the anti-inflammatory topical composition according to this disclosure is between about 3.0 and about 9.0, between about 3.0 and about 8.5, between about 3.0 and about 8.0, between about 3.0 and about 7.5, between about 3.0 and about 7.0, between about 3.0 and about 6.5, between about 3.0 and about 6.0, between about 3.0 and about 5.5, between about 3.0 and about 5.0, between about 3.0 and about 4.5, between about 3.5 and about 9.0, between about 3.5 and about 8.5, between about 3.5 and about 8.0, between about 3.5 and about 7.5, between about 3.5 and about 7.0, between about 3.5 and about 6.5, between about 3.5 and about 6.0, between about 3.5 and about 5.5, between about 3.5 and about 5.0. Between approximately 3.5 and approximately 4.5, between approximately 4.0 and approximately 9.0, between approximately 4.0 and approximately 8.5, between approximately 4.0 and approximately 8.0, between approximately 4.0 and approximately 7.5, between approximately 4.0 and approximately 7.0, between approximately 4.0 and approximately 6.5, between approximately 4.0 and approximately 6.0, between approximately 4.0 and approximately 5.5, between approximately 4.0 and approximately 5.0, between approximately 4. Any range or value between 5 and about 9.0, between about 4.5 and about 8.5, between about 4.5 and about 8.0, between about 4.5 and about 7.5, between about 4.5 and about 7.0, between about 4.5 and about 6.5, between about 4.5 and about 6.0, between about 4.5 and about 6.0, between about 4.5 and about 5.5, between about 4.5 and about 5.0, or between these ranges or values.

[0169] In some embodiments, the pH of the anti-inflammatory topical composition according to this disclosure may be about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8. Approximately 5.9, approximately 6.0, approximately 6.1, approximately 6.2, approximately 6.3, approximately 6.4, approximately 6.5, approximately 6.6, approximately 6.7, approximately 6.8, approximately 6.9, approximately 7.0, approximately 7.1, approximately 7.2, approximately 7.3, approximately 7.4, approximately 7.5, approximately 7.6, approximately 7.7, approximately 7.8, approximately 7.9, approximately 8.0, approximately 8.1, approximately 8.2, approximately 8.3, approximately 8.4, approximately 8.5, approximately 8.6, approximately 8.7, approximately 8.8, approximately 8.9, approximately 9.0, or any range or value between them.

[0170] In some embodiments, the local composition has an acidic pH, such as less than about 7, less than about 6.5, less than about 6.0, less than about 5.5, less than about 5.0, less than about 4.5, or any range or value between therewith.

[0171] In particular, acidic topical compositions may be beneficial in treating elderly patients with cutis laxity or atopic dermatitis. Skin treated with acidic emulsions (e.g., pH 4) showed increased resistance to oxidative damage, enhanced barrier integrity, and reduced roughness and scaling of the skin surface several weeks after treatment. See, for example, M. et al., “Towards Optimal pH of the Skin and Topical Formulations: From the Current State of the Art to Tailored Products”, 8 Cosmetics 69 (2021); A. Kilic et al., “Skin Acidification With a Water-in-Oil Emulsion (pH 4) Restores Disrupted Epidermal Barrier and Improves Structure of Lipid Lamellae in the Elderly”, 46 Journal of Dermatology 457–65 (2019).

[0172] pH adjuster

[0173] In some embodiments, the local composition may include one or more pH adjusters adapted to adjust the pH of the composition to any of the ranges discussed above. In some embodiments, the pH adjuster may comprise one or more suitable mineral acids (e.g., hydrochloric acid, nitric acid, phosphoric acid, phosphorous acid, sulfuric acid, etc.), carboxylic acids (e.g., citric acid, glycolic acid, lactic acid, maleic acid, malic acid, succinic acid, glutaric acid, benzoic acid, malonic acid, salicylic acid, gluconic acid, etc.), polymeric acids (e.g., linear poly(acrylic) acid and its copolymers, such as maleic acid-acrylic acid copolymer, sulfonic acid-acrylic acid copolymer, and styrene-acrylic acid copolymer), crosslinked polyacrylic acid, poly(methacrylic) acid, carrageenan acid, alginate, etc.), and any combination thereof.

[0174] pH can be increased or made more alkaline by adding any suitable alkaline pH adjuster (e.g., sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, etc.). In some embodiments, one or more pH adjusters may comprise: ammonia; monoalkylamines, dialkylamines, and trialkylamines (e.g., trimethylamine); monoalkylolamines, dialkylolamines, and trialkylolamines (e.g., monoethanolamine, diethanolamine, triethanolamine, isopropanolamine, diisopropanolamine, and triisopropanolamine); alkali metal and alkaline earth metal hydroxides (e.g., sodium hydroxide, potassium hydroxide, lithium hydroxide, etc.); alkali metal and alkaline earth metal silicates; and other pH adjusters (e.g., aminomethylpropanol (AMP-95), tetrahydroxypropylethylenediamine, etc.). C-25 (PEG-15 cocoaamine, etc.).

[0175] In some embodiments, one or more pH adjusters comprise buffers. A buffer is one or more chemical compounds added to a solution to allow the solution to resist pH changes caused by dilution or the addition of small amounts of acid or base. An effective buffering system employs a solution containing a large and approximately equal concentration of a conjugate acid-base pair (or buffer). Buffers used herein can be any pharmaceutically acceptable chemical compound, including but not limited to salts of phosphoric acid and citrate (conjugate acid and / or base). In some aspects, the buffer comprises phosphate-buffered saline (PBS) or an alternative phosphate buffer.

[0176] Other ingredients

[0177] The topical compositions according to this disclosure may contain a wide range of optional ingredients or additives. Various non-limiting cosmetic and pharmaceutical ingredients commonly used in the skincare industry for the topical compositions of this disclosure are described by reference to the CTFA International Cosmetic Ingredient Dictionary, 15th edition, 2014, which is incorporated herein by reference in its entirety. Non-limiting examples of such genus of ingredients include: abrasives, anti-acne agents, anti-caking agents (e.g., silica, distarch phosphate, etc.), binders, biological additives, fillers, chelating agents (e.g., disodium EDTA), chemical additives, colorants, cosmetic astringents, cosmetic bactericides, denaturants, pharmaceutical astringents, topical analgesics, film-forming agents, fragrance components, emulsifiers, plasticizers, preservatives (e.g., phenoxyethanol, ethylhexylglycerin, etc.), propellants, reducing agents, skin bleaching agents, skin conditioning agents, skin protectants, etc. Solvents (e.g., ethanol, 1,2-hexanediol, etc.), foam enhancers, water-soluble agents, solubilizers, suspending agents (non-surfactants), sunscreens, UV absorbers and thickeners (aqueous and non-aqueous), solubilizers, chelating agents and exfoliating agents, plant extracts, dipeptides, tripeptides (e.g., tripeptide-1) and their derivatives, tetrapeptides (e.g., tetrapeptide-2 and their derivatives), hexapeptides (e.g., hexapeptide-11, hexapeptide-12, hexapeptide-38 and their derivatives), octapeptides (e.g., octapeptide-45 and their derivatives), and any combination thereof.

[0178] When present, such additional components are preferably present in relatively low concentrations, such as 0.001 wt.% to about 5 wt.%, about 0.001 wt.% to about 1 wt.%, about 0.001 wt.% to about 0.5 wt.%, about 0.001 wt.% to about 0.1 wt.%, about 0.001 wt.% to about 0.05 wt.%, about 0.001 wt.% to about 0.01 wt.%, or any range or value between therewith.

[0179] water

[0180] The partial compositions according to this disclosure may contain water in concentrations of about 0 wt.% to about 98 wt.%, about 5 wt.% to about 95 wt.%, about 10 wt.% to about 90 wt.%, about 15 wt.% to about 85 wt.%, about 20 wt.% to about 80 wt.%, about 25 wt.% to about 75 wt.%, about 30 wt.% to about 70 wt.%, about 35 wt.% to about 65 wt.%, or about 40 wt.% to about 60 wt.%, or any range or value between these concentrations relative to the total weight of the composition. In some embodiments, water is present at a concentration of about 5 wt.%, about 10 wt.%, about 15 wt.%, about 20 wt.%, about 25 wt.%, about 30 wt.%, about 35 wt.%, about 40 wt.%, about 45 wt.%, about 50 wt.%, about 55 wt.%, about 60 wt.%, about 65 wt.%, about 70 wt.%, about 75 wt.%, about 80 wt.%, about 85 wt.%, about 90 wt.%, about 95 wt.%, about 96 wt.%, about 97 wt.%, about 98 wt.%, about 99 wt.%, or any range or value between thereof, relative to the total weight of the composition.

[0181] Composition form

[0182] The topical compositions according to this disclosure can be in any galenical form that ensures the composition is stable, non-irritating to the skin, non-drying to the skin, and / or pleasant and easy to apply. In some embodiments, the topical composition is an emulsion (e.g., an oil-in-water emulsion or an oil-in-water emulsion), gel, cream, cream-gel, solution, suspension, lotion, emulsion, ointment, cream, foam (e.g., an aerosol or self-foaming composition), balm, paste, or sachet. In some embodiments, the topical composition is an emulsion. In some embodiments, the topical composition is a gel. In some embodiments, the topical composition is a cream. In some embodiments, the topical composition is a galenical form suitable for a pump dispenser. In some embodiments, the anti-inflammatory composition is formulated for no-rinse (rather than "rinse-off") application. In some embodiments, the topical composition is an oil-in-water (O / W) emulsion.

[0183] How to use

[0184] In one aspect, this disclosure relates to a method of treating skin affected by skin laxity, the method comprising: applying a topical composition according to this disclosure to the affected skin.

[0185] In one aspect, this disclosure relates to a method for increasing fibroblast turnover in mammalian skin, the method comprising: applying a topical composition according to this disclosure to the mammalian skin. In any of the embodiments described herein, mammalian skin may comprise human skin.

[0186] In one aspect, this disclosure relates to a method for reducing the number of senescent cells in the dermis of a mammal, the method comprising: applying a topical composition according to this disclosure to the mammalian skin. In some embodiments, the method comprises reducing the number of senescent cells in the dermis and removing the senescent cells from the dermis to the epidermis.

[0187] In one aspect, this disclosure relates to a method for reducing the activity of senescent cells in mammalian skin, the method comprising: applying a topical composition according to this disclosure to the mammalian skin. In some embodiments, the method comprises reducing the expression of p16 and / or reducing the activity of said p16 in mammalian skin.

[0188] In one aspect, this disclosure relates to a method for reducing the aging-associated secretory phenotype (SASP) in mammalian epidermal cells, the method comprising: applying a topical composition according to this disclosure to the mammalian skin. In some aspects, this disclosure relates to a method for modulating at least one SASP-related gene in mammalian skin, the method comprising: applying a topical composition according to this disclosure to the mammalian skin, wherein the at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C. In some embodiments, the modulation comprises downregulating one or more genes and / or upregulating one or more genes. In some embodiments, the modulation comprises downregulating one or more genes selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C. In some embodiments, the modulation comprises downregulating IL8. In some embodiments, the modulation comprises downregulating IL1B. In some embodiments, the modulation comprises downregulating HIST1H2BG. In some embodiments, the modulation comprises downregulating UBE2C. In some embodiments, the modulation is performed in keratinocytes.

[0189] In one aspect, this disclosure relates to a method of modulating one or more inflammation-related genes in mammalian skin, the method comprising: applying a topical composition according to this disclosure to mammalian skin, wherein the at least one inflammation-related gene is selected from the group consisting of MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2. In some embodiments, the modulation comprises downregulating one or more genes and / or upregulating one or more genes. In some embodiments, the modulation comprises downregulating one or more genes selected from the group consisting of MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2. In some embodiments, the modulation comprises downregulating MMP2. In some embodiments, the modulation comprises downregulating HSPA8. In some embodiments, the modulation comprises downregulating CSF2RA. In some embodiments, the modulation comprises downregulating CXCL1. In some embodiments, the modulation comprises downregulating IL2RB. In some embodiments, the modulation comprises downregulating NFKBIA. In some embodiments, the modulation comprises downregulating PTGS2. In some embodiments, the gene is associated with skin inflammation. In some embodiments, the modulation is performed in keratinocytes.

[0190] In one aspect, this disclosure relates to a method of regulating the expression of one or more genes associated with p15 activity in mammalian skin, the method comprising: applying a topical composition according to this disclosure to mammalian skin, wherein the one or more genes associated with p15 activity comprise CDKN2B. In some embodiments, the regulation is upregulation. In some embodiments, the method comprises upregulating the expression of CDKN2B in mammalian skin. In some embodiments, the regulation is performed in keratinocytes.

[0191] In one aspect, this disclosure relates to a method for regulating the expression of at least one cell cycle gene in mammalian dermal fibroblasts, the method comprising: applying a topical composition according to this disclosure to mammalian skin, wherein the at least one cell cycle gene in the mammalian fibroblasts is selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH. In some embodiments, regulation comprises downregulating one or more genes and / or upregulating one or more genes. In some embodiments, the regulation includes upregulating one or more genes selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH. In some embodiments, the regulation includes upregulating TK1. In some embodiments, the regulation includes upregulating NDC80. In some embodiments, the regulation includes upregulating HMMR. In some embodiments, the regulation includes upregulating KIF2C. In some embodiments, the regulation includes upregulating UBEC. In some embodiments, the regulation includes upregulating LMNB1. In some embodiments, the regulation includes upregulating CABLES1. In some embodiments, the regulation includes upregulating MCM10. In some embodiments, the regulation includes upregulating H2AFX. In some embodiments, the regulation includes upregulating CASC5. In some embodiments, the adjustment includes increasing ESCO2. In some embodiments, the adjustment includes increasing ERCC6L. In some embodiments, the adjustment includes increasing BUB1. In some embodiments, the adjustment includes increasing BIRC5. In some embodiments, the adjustment includes increasing CCNE2. In some embodiments, the adjustment includes increasing GTSE1. In some embodiments, the adjustment includes increasing CDCA8. In some embodiments, the adjustment includes increasing WEE1. In some embodiments, the adjustment includes increasing CDC45. In some embodiments, the adjustment includes increasing CDK1. In some embodiments, the adjustment includes increasing CCNB2. In some embodiments, the adjustment includes increasing CLSPN. In some embodiments, the adjustment includes increasing BUB1B. In some embodiments, the adjustment includes increasing PKMYT1. In some embodiments, the adjustment includes increasing KIF23.In some embodiments, the adjustment includes increasing TPX2. In some embodiments, the adjustment includes increasing KIF20A. In some embodiments, the adjustment includes increasing NCAPH.

[0192] In one aspect, this disclosure relates to a method for downregulating at least one AP-1 complex transcription factor, the method comprising: applying a topical composition of the present disclosure to mammalian skin. In some embodiments, the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

[0193] In one aspect that can be combined with any other aspect or embodiment, this disclosure relates to a method for activating cell cycle progression, the method comprising: applying a topical composition of the present disclosure to mammalian skin, wherein the composition upregulates CCNE2, CCNB1, or a combination thereof.

[0194] In one aspect, this disclosure relates to a method for increasing the turnover rate of dermal fibroblasts in mammalian skin, the method comprising: applying a topical composition according to this disclosure to the affected skin.

[0195] In one aspect, this disclosure relates to a method of stimulating collagen production in mammalian skin, the method comprising: applying a topical composition according to this disclosure to the affected skin.

[0196] In any aspect or embodiment, the regulated (upregulated or downregulated) gene expression can be measured as a fold change in expression relative to baseline expression or in untreated cells. In some embodiments, upregulation is at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, at least 2.0-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, at least 3.0-fold, at least 3.1-fold, at least 3.2-fold, at least 3.3-fold, at least 3.4-fold, at least 3.5-fold, at least 3.6-fold, at least... 3.7 times, at least 3.8 times, at least 3.9 times, at least 4.0 times, at least 4.1 times, at least 4.2 times, at least 4.3 times, at least 4.4 times, at least 4.5 times, at least 4.6 times, at least 4.7 times, at least 4.8 times, at least 4.9 times, at least 5.0 times, at least 5.5 times, at least 6.0 times, at least 6.5 times, at least 7.0 times, at least 7.5 times, at least 8.0 times, at least 8.5 times, at least 9.0 times, at least 9.5 times, at least 10.0 times, at least 15 times, at least 20 times or more, or any range or value between these values.

[0197] In some embodiments, the reduction is to at least -0.1 times, at least -0.2 times, at least -0.3 times, at least -0.4 times, at least -0.5 times, at least -0.6 times, at least -0.7 times, at least -0.8 times, at least -0.9 times, at least -1.0 times, at least -1.1 times, at least -1.2 times, at least -1.3 times, at least -1.4 times, at least -1.5 times, at least -1.6 times, at least -1.7 times, at least -1.8 times, at least -1.9 times, at least -2.0 times, at least -2.1 times, at least -2.2 times, at least -2.3 times, at least -2.4 times, at least -2.5 times, at least -2.6 times, at least -2.7 times, at least -2.8 times, at least -2.9 times, at least -3.0 times, at least -3.1 times, up to At least -3.2 times, at least -3.3 times, at least -3.4 times, at least -3.5 times, at least -3.6 times, at least -3.7 times, at least -3.8 times, at least -3.9 times, at least -4.0 times, at least -4.1 times, at least -4.2 times, at least -4.3 times, at least -4.4 times, at least -4.5 times, at least -4.6 times, at least -4.7 times, at least -4.8 times, at least -4.9 times, at least -5.0 times, at least -5.5 times, at least -6.0 times, at least -6.5 times, at least -7.0 times, at least -7.5 times, at least -8.0 times, at least -8.5 times, at least -9.0 times, at least -9.5 times, at least -10.0 times, at least -15 times, at least -20 times or more, or any range or value between these values.

[0198] The topical compositions according to this disclosure can be used with a variety of treatment regimens. In some instances, the topical compositions described herein are applied once daily, twice daily, three times daily, or more frequently. In some instances, the topical compositions described herein are applied twice daily. In some embodiments, the topical compositions described herein are applied daily, once daily, every other day, five days a week, once a week, every other week, for two weeks a month, for three weeks a month, once a month, twice a month, three times a month, or more frequently. In some embodiments, the topical compositions described herein are applied twice daily (e.g., in the morning and evening).

[0199] In some embodiments, the topical composition described herein is applied for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years, 4 years, 5 years, 10 years, or longer. In some embodiments, the topical composition described herein is applied twice daily for at least or about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or longer. In some embodiments, the topical composition described herein is applied once daily, twice daily, three times daily, four times daily, or more than four times daily for at least or about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or longer.

[0200] Example

[0201] Example 1. Preparation of topical compositions

[0202] Exemplary partial compositions according to this disclosure are shown in Table 1 below.

[0203] Table 1. Exemplary Topical Compositions

[0204]

[0205]

[0206] Prepare the topical compositions according to Table 1 according to the following general procedure:

[0207] In the main tank, a gelling phase (phase A) is prepared by adding a gelling agent (e.g., acrylate / C10-30 alkyl acrylate crosspolymer), disodium EDTA, glycerol, and water under stirring until completely dispersed. In a first auxiliary tank, an oil phase (phase B) is prepared by adding an oil (e.g., sunflower oil, hydrogenated vegetable oil) and a non-liquid fatty substance (e.g., cetearyl alcohol, synthetic beeswax) under stirring, followed by the addition of an emulsifier (e.g., methylglucose sesquistearate) and an antioxidant (e.g., tocopheryl acetate), and stirring until completely dispersed. Phase B is added to phase A under stirring, followed by the addition of an emulsifying phase, phase C (potassium cetyl phosphate), until an oil-in-water emulsion is observed. Next, the acrylate is neutralized by adding sodium hydroxide solution (e.g., pH about 5-6). Then, a preservative phase, phase F (e.g., phenoxyethanol, ethylhexylglycerin), is added under stirring until completely dispersed.

[0208] In the second auxiliary tank, the liposome mixture (e.g., PRO-LIPO) is stirred. TMThe active liposome phase (G phase) is prepared by mixing NEO, water, and Centella asiatica leaf extract (e.g., HETEROSIDES) until the Centella asiatica leaf extract is completely dispersed in the liposome mixture. The G phase is then added to the pH-adjusted emulsion in the master tank under stirring until completely dispersed. Finally, the AHA phase (H phase) containing mandelic acid is added to the master tank under stirring until completely dispersed and the pH is between 4 and 5, to obtain an oil-in-water emulsion containing liposome-encapsulated Centella asiatica leaf extract and mandelic acid.

[0209] Example 2. Cytotoxicity study of mandelic acid and Centella asiatica extract

[0210] To determine the appropriate dosage of Centella asiatica extract and mandelic acid, human primary adult dermal fibroblasts and human primary adult dermal keratinocytes were purchased from ZenBio (https: / / www.zen-bio.com / , Durham, NC) and plated in the culture medium recommended by ZenBio according to the provided ZenBio protocol.

[0211] Approximately 10,000 cells were seeded per well in a 96-well plate. A 12-point dilution series of Centella asiatica extract or mandelic acid was performed in triplicate of the wells. At 2x step, the concentration of Centella asiatica extract was varied from 0.00025% to 0.25%. At 2x step, the concentration of mandelic acid was varied from 0.00004% to 0.08%. After 24 hours of treatment, an alpha blue assay was performed. The plate was then read in an Envision platereader with 560 nm excitation and 590 nm emission filters to determine cell viability. Fluorescence intensity values ​​were plotted, and these values ​​were recorded in [the following text is incomplete and requires further context to translate accurately]. Figure 1A (Centella asiatica, fibroblasts) Figure 1B (Centella asiatica, keratinocytes) Figure 1C (Mandolic acid, fibroblasts) Figure 1D (Mandelic acid, keratinocytes) is shown. The arrow indicates the highest concentration at which cytotoxicity was not observed in the Almarin assay, in which cells were directly exposed to the respective ingredient in the absence of other topical formulation ingredients. Based on the surprising cellular responses at the tested concentrations, the inventors investigated whether negative effects (e.g., inflammation) could be reduced while providing therapeutic effects (e.g., reduced cellular senescence, increased cell turnover, etc.) by using Centella asiatica extract and mandelic acid (at concentrations lower than those conventionally recommended for their topical application) at concentrations lower than those typically recommended for their topical use.

[0212] Example 3. In vitro analysis of gene expression in human keratinocytes and dermal fibroblasts.

[0213] A series of in vitro gene expression analyses were performed to evaluate the effects of the topical composition according to this disclosure on inflammation, cell turnover, and aging. (See also...) Figure 2 (The figure is a schematic diagram of biomarkers associated with inflammation, cell turnover, and aging). The regulation of gene expression in the presence of Centella asiatica extract, mandelic acid, and a combination of Centella asiatica extract and mandelic acid was tested to assess the effect on genes associated with the aging-associated secretory phenotype (SASP), a phenotype associated with senescent cells that secrete high levels of inflammatory cytokines, immunomodulators, growth factors, and proteases. Human dermal fibroblasts and keratinocytes were grown to convergence and treated for 72 hours with Centella asiatica extract (0.005%), mandelic acid (0.05%), or a combination of both active ingredients (at the same concentration). (The selected concentrations of mandelic acid and Centella asiatica extract were determined to be the highest concentrations that would not cause cytotoxicity, as discussed in Example 2). RNA was extracted and sequenced. Differentially expressed genes were identified by comparison with untreated cells. Genes were annotated using the Reactome database.

[0214] like Figures 3A-3D As shown in the study, multiple SASP-related genes that increase inflammation were downregulated by Centella asiatica extract, indicating a protective effect of Centella asiatica extract on senescent cells. Furthermore, the combination of Centella asiatica extract and mandelic acid synergistically downregulated IL-8 expression. Figure 3A ), IL1B ( Figure 3B ), HIST1H2BG ( Figure 3C ) and UBE2C ( Figure 3D SASP-related genes.

[0215] In addition, such as Figure 4A-4G As shown, pro-inflammatory gene expression was significantly downregulated by Centella asiatica extract alone and in combination with mandelic acid, particularly for MMP2 (… Figure 4A HSPA8 Figure 4B ), CSF2RA Figure 4C ), CXCL1 Figure 4D ), IL2RB ( Figure 4E ), NFKBIA Figure 4F ) and PTG52 ( Figure 4G This is especially true. Furthermore, the combination of Centella asiatica extract and mandelic acid synergistically downregulated the expression of genes associated with general inflammation, and this downregulation may be related to the downregulation of SASP-related genes. The data are summarized in Table 2.

[0216] Table 2. Regulation of gene expression (fold change) by Centella asiatica extract, mandelic acid, and combinations of Centella asiatica extract and mandelic acid.

[0217] markers Centella asiatica Mandelic acid Centella Asiatica + Mandelic Acid IL8 -5.50 -1.46 -7.17 IL1B -5.05 1.73 -6.41 TNF -8.62 -2.69 -7.83 HIST1H4H -4.82 -0.54 -4.16 HIST1H2BG -3.98 0.45 -4.68 UBE2C -2.77 0.51 -5.44 UBB -5.06 -3.87 -5.69 UBC -3.43 -1.09 -3.16 CEBPB -4.63 -2.45 -3.69 HIST1H2BD -4.05 0.55 -4.02 HIST1H4E -11.59 0.60 -6.62 HIST1H3D -11.21 -0.23 -11.00

[0218] like Figure 5 As shown, both individual and combined Centella asiatica extracts and mandelic acid upregulate p15 (CDKN2B) expression, which involves evading oxidative stress-induced senescence due to the defective binding of p16INK4A to CDK4. Thus, treatment with the compositions according to this disclosure can prevent cellular senescence of keratinocytes and reduce the presence of senescent cells in the dermis.

[0219] Now for reference Figure 6 The combination of Centella asiatica extract and mandelic acid synergistically and significantly upregulated the expression of cell cycle-related genes TK1, UBEC, H2AFX, BUB1, CDCA8, CCNB2, KIF23, NDC80, LMNB1, CASC5, BIRC5, WEE1, CLSPN, TPX2, HMMR, CABLES1, ESCO2, CCNE2, CDC45, BUB1B, KIF20A, KIF2C, MCM10, ERCC6L, GTSE1, CDK1, PKMYT1, and NCAPH in dermal fibroblasts. Significantly, only the combination of Centella asiatica extract and mandelic acid significantly upregulated all of the genes shown (y-axis shows the fold change in log2). The data suggest increased cell turnover and consequently reduced cellular senescence.

[0220] Additionally, now refer to Figures 7A-7J The combination of Centella asiatica extract and mandelic acid synergistically upregulates the p16-interacting gene WEE1. Figure 7A H2AFX Figure 7B ), LMNB1 ( Figure 7C ), CCNB2 ( Figure 7D ), BUB1( Figure 7E ), CDC45 Figure 7F ), CDK1 Figure 7G ), BIRC5 ( Figure 7H ), MCM10 Figure 7I ) and CCNE2 ( Figure 7J The expression of p16 in fibroblasts indicates that the combination counteracts p16 in fibroblasts, thereby reversing cellular senescence.

[0221] Skin laxity is associated with the loss of ECM (collagen, elastin, glucosamine, etc.), leading to thinning, fragility, and vascular leakage of the skin. This breakdown is related to the production of ROS, which stimulate the activation of transcription factors (such as AP-1) and the synthesis of matrix metalloproteinases (MMPs). JunB is a major component of the transcription factor AP-1. Therefore, downregulating this factor will reduce or limit ECM breakdown. Now refer to Figure 8 Fibroblasts treated with a combination of Centella asiatica extract (0.005%) and mandelic acid (0.05%) showed significant downregulation of JunB and FosL2, both of which are transcription factors and members of the AP-1 complex. This downregulation impaired the ability of p21 (CDKN1A) to inhibit cell cycle and, consequently, activate cell cycle progression as demonstrated by the upregulation of cyclin E2 (CCNE2) and cyclin B1d (CCNB1). (All fold changes are significant, where p < 0.05).

[0222] In summary, gene expression data indicate that the combination of Centella asiatica extract and mandelic acid induces a strong synergistic effect in the following aspects: (i) significantly reducing the inflammatory secretory group (SASP) of senescent cells, thereby escaping the senescent cell activity of keratinocytes; (ii) increasing the turnover rate of dermal fibroblasts; and (iii) reducing p16 (a marker of senescence and a cell cycle arrestor), which suggests an overall reduction in fibroblast cellular senescence.

[0223] Example 4. In vitro analysis of cellular senescence activity

[0224] To investigate the effect of the topical composition according to this disclosure on the response of senescent cells when applied topically to the skin, a test composition was prepared according to Example 1 and tested in vitro on discarded human skin samples. Biopsies were taken from the discarded human skin samples, and the tissues were placed in 24-well plates for culture. After adaptation, the skin samples were treated with the test composition once daily for 10 days. The samples were stained with p16 to assess senescence activity using untreated skin samples as a baseline.

[0225] Figures 9A-9DImages of p16-stained skin samples are shown, comparing untreated skin (left) to skin treated with the test composition (right). Cell nuclei appear blue, while p16 appears yellow. Untreated skin shows evidence of p16 (senescent cells) in the dermis. In contrast, the sample treated with the test composition shows a loss of senescent cells in the dermis and evidence of senescent cells in the epidermis. In several images, a high concentration of senescent cells was observed in hair follicles, where cells undergo natural senescence as part of the hair cycle. These senescent cells are expected to be present but were not considered in this analysis.

[0226] In vitro results confirmed the gene expression data associated with p16 senescence activity (Example 2), as the test composition containing Centella asiatica extract and mandelic acid appeared to reduce and even counteract p16 activity. In vitro results also showed a reduced dermal cell turnover rate compared to untreated skin, demonstrated by a decreased population of senescent cells in the treated dermis.

[0227] Furthermore, surprisingly, the compositions according to this disclosure appear to promote the clearance of senescent dermal cells from the dermis to the epidermis. Without being bound by any particular theory, the tested compositions appear to potentially promote the clearance of senescent cells through a signaling mechanism initiated in the epidermis and signaling downwards to senescent dermal cells (possibly due to the collagen-stimulating effect of Centella asiatica extract), thereby reversing dermal aging in the presence of increased epidermal senescent cells and the appearance of terminally differentiated cells in the upper epidermis, and then promoting epidermal turnover (and protection from the effects of these cells). This suggests a form of senescent cell clearance and cell regeneration that would be highly beneficial for the management of skin laxity.

[0228] While it is believed that those skilled in the art will readily understand the foregoing terms, the following definitions are set forth in order to explain the subject matter currently disclosed.

[0229] The terms “a” or “an” can refer to one or more of the entities, i.e., they can refer to multiple indicators. Therefore, the terms “a” or “an,” “one or more,” and “at least one” are used interchangeably herein. Furthermore, the reference to “element” by the indefinite article “a” or “an” does not preclude the possibility of more than one element, unless the context clearly requires the presence of one and only one element.

[0230] Throughout this specification, references to "one embodiment," "an embodiment," "one aspect," or "an aspect" indicate that a particular feature, structure, or characteristic described in connection with an embodiment is included in at least one embodiment of this disclosure. Therefore, the phrases "in one embodiment" or "in one embodiment" appearing throughout this specification do not necessarily refer to the same embodiment. Furthermore, specific features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0231] As used herein, the terms “about” or “approximately” when preceding a numerical value indicate a range of 10% of the value plus or minus the value.

[0232] As those skilled in the art will understand, for any and all purposes, particularly for the purpose of providing a written description, all scopes disclosed herein also encompass any and all possible subscopes and combinations thereof. Any listed scope can be readily recognized as sufficiently describing and implementing an identical scope that can be decomposed into at least two, three, four, five, ten, etc., equal parts. As a non-limiting example, each scope discussed herein can be readily decomposed into a lower third, a middle third, and an upper third, etc. As those skilled in the art will also understand, all language such as “at most,” “at least,” “greater than,” “less than,” etc., includes the listed numbers and refers to a scope that can subsequently be decomposed into subscopes as discussed above. Finally, as those skilled in the art will understand, a scope includes each individual member. Thus, for example, a group having 1-3 cells means a group having 1, 2, or 3 cells. Similarly, a group having 1-5 cells means a group having 1, 2, 3, 4, or 5 cells, and so on.

[0233] Unless otherwise defined, all terms used herein (including technical and scientific terms) shall have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. It should be further understood that terms defined in commonly used dictionaries shall be interpreted as having a meaning consistent with their meaning in the context of this application and related fields, and shall not be interpreted in an idealized or overly formal sense unless expressly defined herein. While not expressly defined below, such terms shall be interpreted according to their ordinary meaning.

[0234] Furthermore, when features or aspects of this disclosure are described in accordance with the Markush Group, those skilled in the art will recognize that this disclosure is also described in accordance with any individual member or subgroup of the Markush Group.

[0235] Unless the context otherwise indicates, it is expressly intended that the various features of the invention described herein may be used in any combination. Furthermore, this disclosure also contemplates that in some embodiments, any feature or combination of features shown herein may be excluded or omitted. For example, if the specification states that a complex comprises components A, B, and C, it is expressly intended that any one or a combination of A, B, or C may be omitted or discarded individually or in any combination.

[0236] Unless otherwise expressly indicated, all specified embodiments, features, and terms are intended to include the listed embodiments, features, or terms and their equivalents.

[0237] All patents, patent applications, provisional applications and publications mentioned or cited herein are incorporated herein by full reference to the extent that they are not inconsistent with the express teachings in this specification, including all figures and tables.

[0238] Reference will now be made in detail to some specific embodiments contemplated in this disclosure. While various embodiments are described herein, it should be understood that they are not intended to limit the invention to the described embodiments. Rather, the invention is intended to cover alternatives, modifications, and equivalents that may be included within the spirit and scope of the technology defined by the appended claims.

Claims

1. A topical composition comprising: Centella asiatica extract, wherein the Centella asiatica extract is encapsulated in liposomes; and α-hydroxy acid (AHA), The pH of the composition is less than 5.

2. The topical composition according to claim 1, wherein the composition is in the form of an emulsion.

3. The topical composition according to claim 2, wherein the emulsion is an oil-in-water (O / W) emulsion.

4. The topical composition according to any one of claims 1 to 3, wherein the pH of the composition is greater than or equal to 4 and less than 5.

5. The topical composition according to any one of claims 1 to 4, wherein the Centella asiatica extract is present at a concentration of at least about 0.01 wt.% and less than 0.2 wt.% relative to the total weight of the composition.

6. The topical composition according to any one of claims 1 to 5, wherein the Centella asiatica extract is present at a concentration of about 0.01 wt.% to about 0.1 wt.% relative to the total weight of the composition.

7. The topical composition according to any one of claims 1 to 6, wherein the Centella asiatica extract comprises a triterpenoid compound.

8. The topical composition according to any one of claims 7, wherein the triterpenoid compound is selected from one or more of the group consisting of: madecassoside, asiaticoside, madecassic acid, and asiatic acid.

9. The topical composition according to claim 7 or claim 8, wherein the triterpenoid compound is present in the Centella asiatica extract at a concentration of at least about 70 wt.% relative to the total weight of the Centella asiatica extract.

10. The topical composition according to any one of claims 7 to 9, wherein the triterpenoid compound is present in the composition at a concentration of at least about 0.005 wt.% and less than 0.2 wt.% relative to the total weight of the composition.

11. The topical composition according to any one of claims 7 to 10, wherein the triterpenoid compound is present in the Centella asiatica extract at a concentration of at least about 0.01 wt.% to about 0.05 wt.% relative to the total weight of the composition.

12. The topical composition according to any one of claims 1 to 11, wherein the AHA comprises mandelic acid.

13. The topical composition according to any one of claims 1 to 12, wherein the AHA is present at a concentration of at least about 0.01 wt.% and less than 0.5 wt.% relative to the total weight of the composition.

14. The topical composition according to any one of claims 1 to 13, wherein the AHA is present at a concentration of about 0.05 wt.% to about 0.3 wt.% relative to the total weight of the composition.

15. The topical composition according to any one of claims 1 to 14, wherein the liposomes are present at a concentration of about 0.03 wt.% to about 4 wt.% relative to the total weight of the composition.

16. The topical composition according to any one of claims 1 to 15, wherein the liposome comprises lecithin, propylene glycol, or a combination thereof.

17. The topical composition according to any one of claims 1 to 16, wherein the liposomes comprise an antioxidant.

18. The topical composition of claim 17, wherein the antioxidant is tocopherol, tocopheryl acetate, or a combination thereof.

19. The topical composition according to any one of claims 1 to 18, wherein the average particle size of the liposomes is 100 nm to 300 nm.

20. The topical composition according to any one of claims 1 to 19, wherein the average particle size of the liposomes is about 250 nm.

21. The topical composition according to any one of claims 1 to 20, wherein the Centella asiatica extract is present in the form of an active solution.

22. The topical composition of claim 21, wherein the liposomes and the active solution are present in a weight ratio of about 1:1 to about 1:

20.

23. The topical composition according to claim 21 or claim 22, wherein the liposomes and the active solution are present in a weight ratio of about 1:2 to about 1:

10.

24. The topical composition according to any one of claims 1 to 23, wherein the Centella asiatica extract and the AHA are present in a weight ratio of about 1:5 to 1:

15.

25. The topical composition according to any one of claims 1 to 24, further comprising an antioxidant, said antioxidant being separate from any antioxidant in said liposomes.

26. The topical composition according to any one of claims 1 to 25, wherein the antioxidant, separate from any antioxidant in the liposomes, is tocopheryl acetate, tocopherol, or a combination thereof.

27. The topical composition according to any one of claims 1 to 26, further comprising at least one of a moisturizer, emollient, or humectant.

28. The topical composition according to any one of claims 1 to 27, further comprising glycerol.

29. The topical composition according to any one of claims 1 to 28, wherein the composition is a cream or emulsion.

30. A topical composition comprising: Centella asiatica extract, wherein the Centella asiatica extract is encapsulated in liposomes; and α-hydroxy acid (AHA), in: The composition is an emulsion; The pH of the composition is 4 to 5; and The composition reduces biomarkers of skin inflammation in mammalian skin cells.

31. A topical composition comprising: α-hydroxy acid (AHA); and Triterpenoids, selected from the group consisting of asiaticoside, asiaticoside, asiaticopic acid, and asiatic acid, wherein the triterpenoids are encapsulated in liposomes. The pH of the composition is 4 to 5.

32. The topical composition of claim 31, wherein the AHA comprises mandelic acid.

33. The topical composition according to claim 31 or claim 32, wherein the AHA is present in the composition at a concentration of at least about 0.01 wt.% and less than 0.5 wt.% relative to the total weight of the composition.

34. The topical composition according to any one of claims 31 to 33, wherein the triterpenoid compound is present in the composition at a concentration of at least about 0.005 wt.% and less than 0.2 wt.% relative to the total weight of the composition.

35. The topical composition according to any one of claims 31 to 34, wherein the composition is an emulsion.

36. The topical composition according to any one of claims 1 to 35, wherein the composition is effective in reducing the number of senescent cells in the dermis of mammals.

37. The topical composition according to any one of claims 1 to 36, wherein the composition is effective in reducing the aging-associated secretory phenotype (SASP) in mammalian epidermal cells.

38. The topical composition according to any one of claims 1 to 37, wherein the composition is capable of effectively downregulating at least one SASP-related gene, said at least one SASP-related gene being selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

39. The topical composition according to any one of claims 1 to 38, wherein the composition is capable of effectively increasing the expression of at least one cell cycle gene in mammalian dermal fibroblasts.

40. The topical composition of claim 39, wherein at least one cell cycle gene in the mammalian fibroblast is selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

41. The topical composition according to any one of claims 1 to 40, wherein the composition is capable of effectively downregulating at least one gene associated with skin inflammation, said at least one gene associated with skin inflammation being selected from the group consisting of: MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2.

42. The topical composition according to any one of claims 1 to 41, wherein the composition effectively downregulates the AP-1 complex transcription factor.

43. The topical composition according to claim 42, wherein the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

44. The topical composition according to any one of claims 1 to 43, wherein the composition can effectively activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

45. The topical composition according to any one of claims 1 to 30, wherein the Centella asiatica extract and the AHA are present at a concentration that can effectively and synergistically downregulate at least one SASP-related gene, wherein the at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG and UBE2C.

46. ​​The topical composition according to any one of claims 1 to 30 or claim 45, wherein the Centella asiatica extract and the AHA are present at a concentration that effectively and synergistically upregulates at least one cell cycle gene in mammalian dermal fibroblasts, said at least one cell cycle gene being selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

47. The topical composition according to any one of claims 1 to 30 or 45 to 46, wherein c at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG and UBE2C.

48. The topical composition according to any one of claims 1 to 30 or 45 to 47, wherein the Centella asiatica extract and the AHA are present at a concentration that can effectively and synergistically downregulate the AP-1 complex transcription factor.

49. The topical composition of claim 48, wherein the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

50. The topical composition according to any one of claims 1 to 30 or 45 to 49, wherein the Centella asiatica extract and the AHA are present at concentrations that can effectively and synergistically activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

51. The topical composition according to any one of claims 31 to 34, wherein the triterpenoid compound and the AHA are present in a manner that effectively and synergistically downregulates the concentration of at least one SASP-related gene, wherein the at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG, and UBE2C.

52. The topical composition according to any one of claims 31 to 34 or 51, wherein the triterpenoid compound and the AHA are present at a concentration capable of effectively and synergistically upregulating at least one cell cycle gene in mammalian dermal fibroblasts, said at least one cell cycle gene being selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A, and NCAPH.

53. The topical composition according to any one of claims 31 to 34 or 51 to 52, wherein the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically downregulate at least one SASP-related gene, wherein the at least one SASP-related gene is selected from the group consisting of IL8, IL1B, HIST1H2BG and UBE2C.

54. The topical composition according to any one of claims 31 to 34 or 51 to 53, wherein the triterpenoid compound and the AHA are present at a concentration that can effectively and synergistically downregulate the AP-1 complex transcription factor.

55. The topical composition according to claim 54, wherein the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

56. The topical composition according to any one of claims 31 to 34 or 51 to 55, wherein the triterpenoid compound and the AHA are present at concentrations that can effectively and synergistically activate cell cycle progression by upregulating CCNE2, CCNB1, or a combination thereof.

57. A method for treating skin affected by cutis laxity, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to the affected skin.

58. A method for increasing fibroblast turnover in mammalian skin, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to the skin of the mammal.

59. A method for reducing the number of senescent cells in the dermis of mammals, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to mammalian skin.

60. A method for reducing the aging-associated secretory phenotype (SASP) in mammalian epidermal cells, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to mammalian skin.

61. A method for downregulating at least one SASP-related gene in mammalian skin, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to the skin of a mammal. The at least one SASP-related gene is selected from the group consisting of: IL8, IL1B, HIST1H2BG, and UBE2C.

62. A method for increasing the expression of at least one cell cycle gene in mammalian dermal fibroblasts, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to the skin of a mammal. At least one cell cycle gene in the mammalian fibroblasts mentioned above is selected from the group consisting of: TK1, NDC80, HMMR, KIF2C, UBEC, LMNB1, CABLES1, MCM10, H2AFX, CASC5, ESCO2, ERCC6L, BUB1, BIRC5, CCNE2, GTSE1, CDCA8, WEE1, CDC45, CDK1, CCNB2, CLSPN, BUB1B, PKMYT1, KIF23, TPX2, KIF20A and NCAPH.

63. A method for downregulating at least one gene associated with skin inflammation, the method comprising: Apply the composition according to any one of claims 1 to 56 to the skin of a mammal. The at least one of the genes associated with skin inflammation is selected from the group consisting of: MMP2, HSPA8, CSF2RA, CXCL1, IL2RB, NFKBIA, and PTGS2.

64. A method for downregulating at least one AP-1 complex transcription factor, the method comprising: Apply the topical composition according to any one of claims 1 to 56 to mammalian skin.

65. The method of claim 64, wherein the AP-1 complex transcription factor is at least one selected from JunB and FosL2.

66. A method for activating cell cycle progression, the method comprising: Applying a topical composition according to any one of claims 1 to 56 to mammalian skin, wherein the topical composition upregulates CCNE2, CCNB1, or a combination thereof.

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