Novel anti-aging compounds

Abstract

The present invention relates to esters of formula (I) or pharmaceutically acceptable salts thereof and their use as medicaments, in particular for the treatment of skin diseases or skin cancer. The present invention further relates to the use of esters of formula (I) or pharmaceutically acceptable salts thereof in cosmetic and nutritional compositions for preventing or reducing the signs of aging in healthy subjects.

Description

[Technical Field]

[0001] The present invention relates to novel esters made from mandelic acid (derivatives) and phenylpropanoic acid (derivatives) and their use as pharmaceuticals, especially for the treatment of skin disorders. The invention further relates to cosmetic and nutritional compositions comprising the novel esters for preventing or reducing the signs of aging in healthy subjects. [Background technology]

[0002] Aging is an inevitable biological process characterized by the gradual deterioration of various physiological functions, making elderly people increasingly weak and susceptible to disease. The aging process is associated with several chronic, degenerative, and inflammatory diseases. It is characterized by the accumulation of macromolecular damage associated with cellular senescence, impaired tissue regeneration, and progressive loss of physiological integrity (Non-Patent Document 1). Senescence is a state in which cells cease proliferation, lose tissue-specific gene expression, and express inflammatory genes, a state known as the senescence-associated secretory phenotype (SASP). Aging is caused by various stresses, the accumulation of DNA damage, and subsequent epigenetic changes (Non-Patent Document 2; Non-Patent Document 1). The dermis is a connective tissue occupied by fibroblasts, which are involved in the synthesis and secretion of matrix components such as collagen and elastin, which maintain skin structure and provide elasticity, resistance, and strength to the tissue (Non-Patent Document 3; Non-Patent Document 4). Human dermal fibroblasts are involved in almost all skin processes by interacting with both the epidermis and other resident dermal cells (such as endothelial cells, nerve cells, inflammatory cells, and adipocytes). Furthermore, signaling from the dermal compartment underlies the maintenance and homeostasis of epidermal stem cells (Non-Patent Document 3; Non-Patent Document 5). Therefore, dermal fibroblasts are the primary target cells for counteracting skin aging.

[0003] A prominent feature of skin aging is the shift to aging gene expression, which accounts for many of the undesirable structural changes in aging skin, resulting in wrinkles, sagging, and atrophy (Non-Patent Document 2). The age-related aging transition is triggered by intrinsic factors such as telomere shortening, as well as extrinsic factors including UV light, air pollution, cigarette smoke, and metabolic stress, all of which cause DNA damage. With aging, dermal fibroblasts acquire an aging phenotype that significantly impacts skin structure and function. This phenotype is characterized by the following changes in gene expression:

[0004] -Lower expression of collagen and elastin genes. Loss of collagen and elastin leads to loss of firmness and the formation of deeper wrinkles (Non-Patent Document 6).

[0005] - Higher expression of matrix metallopeptidases (e.g., MMP1 and MMP3). These enzymes degrade the dermal matrix and are selectively overexpressed in association with traumatic tissue injury to remove damaged tissue. In aging, their overexpression contributes to a decrease in skin integrity, thickness, and elasticity (Non-Patent Document 7).

[0006] -Decreased expression of tissue inhibitor of metalloproteinase 1 (TIMP1), which exhibits potent inhibition of matrix metallopeptidases and is released by fibroblasts to control inflammatory degradation by MMPs. The decrease in TIMP1 expression associated with fibroblast aging contributes to increased skin bond breakdown associated with aging, both ex vivo and in vivo (Non-Patent Document 7).

[0007] - Higher expression of inflammatory mediator genes, which are characteristic of the senescence-associated secretory phenotype (SASP), which, in addition to mediating nonspecific inflammation, plays a paracrine role in the progression of senescence (Non-Patent Document 1).

[0008] -Cyclin-dependent kinase inhibitor genes, especially p21, in fibroblasts cip1 / waf1 (CDKN1A) and p16 in keratinocytesINK4a Higher expression of CDKN2A (CDKN2A). Overexpression in senescence causes cell cycle arrest, resulting in non-proliferating and inactive cells (Non-Patent Document 8).

[0009] At the cellular level, the transition to senescence is driven by epigenetic modifications, and the accumulation of such changes plays an essential role in the transition to the senescent phenotype (Non-Patent Document 2).

[0010] Lower expression of DNA methyltransferase 1 (DNMT1) is characteristic of aging dermal fibroblasts and keratinocytes. DNMT1 is known as a "maintenance DNMT" because it preserves the original methylation pattern during cell division. DMNT1 expression is inversely correlated with CDKN1A expression and chronological age in human skin samples and cultured fibroblasts (Non-Patent Document 9).

[0011] - Lower expression of class III histone deacetylases (HDACs), particularly SIRT1, which mediate cell survival, UV damage response, DNA repair, and tissue regeneration, as well as energy metabolism, inflammation, and oxidative stress control (Non-Patent Document 10). In dermal fibroblasts, SIRT1 expression significantly decreases with age (Non-Patent Document 11; Non-Patent Document 12), and upregulation or downregulation of SIRT1, in particular, delays or accelerates fibroblast aging, respectively (Non-Patent Document 13).

[0012] While the pharmaceutical industry is increasingly developing new drugs to counter cellular aging, the development of truly anti-aging skin care products is hindered by the requirement for active cosmetic ingredients with well-established safety profiles. The ban on animal testing of cosmetic ingredients in the EU and some US states primarily limits the chemical scope of new cosmetic ingredients to those naturally occurring chemicals that are part of the human diet or metabolism. While some dietary compounds, such as resveratrol and curcumin, have been associated with anti-aging properties, their effects are related only to the potential delay of aging. Such compounds have not been shown to reverse aging or to rejuvenate aging tissues. Patent Document 1 discloses a skin aging inhibitor containing resveratrol 3-O-α-glucoside for preventing skin aging symptoms such as age spots, dullness, wrinkles, sagging skin, and roughness. Patent Document 2 discloses a composition containing phosphorylated resveratrol for treating and reducing the symptoms of skin aging.

[0013] Unlike resveratrol and curcumin, the esters of the present invention have been found to not only slow but reverse aging, making them suitable for the treatment of many inflammatory diseases or disorders, as well as cosmetic products for promoting skin health and preventing the signs of skin aging.

[0014] Furthermore, the esters of the present invention have been found to have anticancer properties, for example, by enhancing DNA damage repair, as demonstrated in two models in which skin cancer-associated CPDs (cyclobutane pyrimidine dimers) were induced in human skin by ultraviolet (UV) irradiation. Skin cancer is cancer that originates in the skin and is also known as cutaneous neoplasia. Skin cancer is the most common form of cancer, accounting for at least 40% of cancer cases worldwide (Non-Patent Document 14). There are many types of skin cancer, including basal cell carcinoma, squamous cell carcinoma, melanoma, and Merkel cell carcinoma. The first two, along with many less common skin cancers, are known as non-melanoma skin cancers. Basal cell carcinoma grows slowly and can damage surrounding tissue, but is unlikely to spread to distant areas or lead to death. It often appears as a painless, raised area of ​​skin that may have a sheen or be accompanied by small blood vessels, or it may present as a raised area with ulcers. Squamous cell skin cancer is more likely to spread. It usually appears as a hard lump with a scaly top, but may also ulcerate. Melanoma is the most aggressive. Signs include moles that vary in size, shape, or color; have irregular borders; have two or more colors; are itchy, or bleed. The most common type is nonmelanoma skin cancer, which affects at least 2 to 3 million people annually. Of nonmelanoma skin cancers, approximately 80% are basal cell carcinoma and 20% are squamous cell carcinoma.

[0015] Ultraviolet radiation from sunlight is thought to be the primary cause of skin cancer. Environmental carcinogens (environmental pollutants) can also cause skin cancer. Examples of environmental carcinogens can include contaminated drinking water, poor indoor air quality, chemical pollutants (e.g., asbestos), food chemicals (e.g., dioxin), and ionizing radon radiation.

[0016] Although any area of ​​the body can be affected by skin cancer, it typically occurs on the face, neck, head, and arms. Basal cell carcinoma and squamous cell carcinoma are responsible for approximately 2,700 deaths annually in the United States, and melanoma is responsible for approximately 7,400 deaths annually (15).

[0017] In the case of basal cell carcinoma or squamous cell carcinoma, several options may be given, including surgery, local chemotherapy, photodynamic therapy, or radiation therapy.In the case of melanoma, treatment may include surgery, chemotherapy, isolated limb perfusion, immunotherapy, and radiation therapy.However, some of these treatments have many drawbacks, such as flu-like symptoms, extreme fatigue, hair loss, DNA damage, the development of secondary cancers, radiation burns to the skin, and cell migration into the bloodstream.In addition, there is no treatment available for the safe long-term prevention or safe prevention of remission of skin cancer.Therefore, there is a need in the art for new and effective skin cancer treatments. [Prior art documents] [Patent documents]

[0018] [Patent Document 1] U.S. Patent No. 10,149,809 [Patent Document 2] U.S. Patent No. 846,5973 [Non-patent literature]

[0019] [Non-Patent Document 1] Lee et al. 2021 [Non-patent document 2] Orioli et al. 2018 [Non-patent document 3] Murphree 2017 [Non-patent document 4] Arseni et al2018 [Non-Patent Document 5] Sriram et al. 2015 [Non-patent document 6] Ezure et al. 2019 [Non-Patent Document 7] Hornebeck et al., 2003 [Non-patent document 8] Idda et al. 2020 [Non-Patent Document 9] Sen et al., 2010 [Non-Patent Document 10] Garcia-Peterson et al. 2017 [Non-Patent Document 11] Tigges et al. 2014 [Non-Patent Document 12] Carlomosti et al., 2017 [Non-Patent Document 13] De Cabo et al., 2015 [Non-Patent Document 14] Cakir et al. 2012 [Non-Patent Document 15] Aggarwal et al. 2021 Summary of the Invention [Problem to be solved by the invention]

[0020] The present inventors have found that the compounds according to the present invention have anti-aging properties. More specifically, the compounds of the present invention can inhibit age-related gene expression in human fibroblasts (i.e., prevent the formation of a senescent cell state), and even more interestingly, the compounds can revert senescent cells to a non-senescent state. The present invention is described in the claims. [Means for solving the problem]

[0021] Detailed Description of the Invention Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0022] The esters of the present invention are obtained by formal condensation between the carboxylic acid in 3-phenylpropanoic acid (or a derivative thereof) and the α-hydroxy group in 2-hydroxy-2-phenylacetic acid (or a derivative thereof). In this context, "2-hydroxy-2-phenylacetic acid" (IUPAC name) may be simply referred to as "mandelic acid". Similarly, "3-phenylpropanoic acid" (IUPAC name) may be simply referred to as "phenylpropanoic acid". The prefixes "(R)" and "(S)" have their usual meaning in the art and indicate the (R)-enantiomer or the (S)-enantiomer, respectively. Similarly, the prefix "(RS)" refers to a racemic mixture.

[0023] In this context, "non-therapeutic benefit" refers to the effect of the esters in the cosmetic or nutritional compositions of the present invention to maintain, reduce, or enhance a physiological process or parameter within a normal physiological range in a healthy subject. Non-limiting examples of non-therapeutic benefits are the inhibition or reversal of cellular aging, the inhibition or reversal of aging, the reduction of facial wrinkles, the maintenance or improvement of skin tone, the maintenance or improvement of muscle endurance or strength, the optimization of sports endurance, the maintenance or improvement of mitochondrial function, and the counteraction or signs of aging.

[0024] In this context, the term "treatment" should be understood in the broadest sense as the prevention, improvement, or treatment of a disease or disorder. Accordingly, treatment is also intended to include prophylactic treatment of a disease or disorder. Thus, "treatment" refers to a therapeutic effect (i.e., medical or therapeutic benefit) in a disease or disorder that results in the reduction, alleviation, mitigation, or reduction of at least one clinical symptom associated with the disease or disorder, or the delay in the progression of the disease or disorder, and / or the prevention or delay in the onset of the disease or disorder. Thus, treatment should be understood as the effect of a pharmaceutical on a pathological process with the purpose of preventing or counteracting a disease or disorder, or a physiological process that potentially leads to a disease or disorder. Non-limiting examples of therapeutic effects include the inhibition or reversal of disease-related cellular senescence, the improvement of mitochondrial biogenesis and function, the reduction or prevention of inflammation, the relief of pain, the improvement of wound healing, the inhibition or reversal of disease-related cellular malignancy, the inhibition of malignant cell proliferation, and cytotoxicity against malignant cells.

[0025] The term "effective amount" refers to an amount of the ester of the present invention sufficient to produce the desired effect. The effective amount will vary depending on the application for which the composition is being used, the age and physical condition of the subject, the severity of the disease or disorder, the duration of treatment, the nature of any concurrent treatment, the carrier used, and similar factors within the knowledge and expertise of those skilled in the art.

[0026] The term "support" is used interchangeably with the terms "maintain," "restore," or "preserve." The term "reduce" is used interchangeably with the terms "lower," "counteract," "diminish," or "reduce." The term "normalize" is used interchangeably with the terms "modulate" or "regulate." The term "improve" is used interchangeably with the terms "enhance," "promote," "stimulate," "increase," or "elevate."

[0027] In this context, it should be understood that the esters of the present invention may be in the form of pharmaceutically acceptable salts. Suitable examples can be found, for example, in Remington's Pharmaceutical Sciences, 17th Edition. Similarly, various solvates of the esters or their pharmaceutically acceptable salts are also within the scope of the present invention.

[0028] In the present context, it is to be understood that alkyl and alkoxy groups may be linear, branched, or cyclic. As a non-limiting example, a C3-alkyl may be linear n-propyl (-CH2CH2CH3), isopropyl (-CH(CH3)2), or cyclopropyl. Most preferably, the alkyl and / or alkoxy groups are linear or branched.

[0029] The present invention will now be described in more detail. It should be understood that the present invention may be embodied in different forms and should not be construed as limited to the embodiments set forth herein.

[0030] The present inventors have found that the compound of formula (I) has anti-aging properties. More specifically, the compound of the present invention can inhibit aging-related gene expression in human fibroblasts (i.e., prevent the formation of senescent cells), and even more interestingly, the compound can revert senescent cells to a non-senescent state. These observations make the compound suitable for the treatment of various diseases and disorders, particularly skin diseases, rheumatic diseases, and wounds. Furthermore, this finding makes the compound suitable for use in cosmetic and nutritional compositions to prevent the signs and symptoms of aging in healthy subjects.

[0031] In a first aspect, the present invention relates to compounds of general formula (I) or a pharmaceutically acceptable salt thereof: [ka] During the ceremony, * denotes the (S) or (R) enantiomer or any mixture thereof; z is an integer of 0, 1, 2, 3, 4, or 5; R 1 is independently selected from F, OH, and C1-C6 alkoxy; n is an integer of 0, 1, 2, 3, 4, or 5; R 2 is independently selected from F, OH, and C1-C6 alkoxy; R 3 is selected from H and C1-C8 alkyl.

[0032] (R 1 ) number of radicals (z) The inventors have discovered that compounds according to the present invention may contain up to five (i.e., z=5) (R 1 ) substituent. In one embodiment, z is an integer of 0 to 4, preferably z is an integer of 0 to 3, more preferably z is an integer of 0 to 2, and most preferably z is an integer of 1 to 3. Thus, it is most preferred that the phenylpropanoic acid portion of the ester has one or two substituents.

[0033] (R 2 ) Number of groups (n) The inventors have discovered that compounds according to the present invention can be used in combination with up to five (i.e., n=5) (R 2 In one embodiment, n is an integer from 0 to 4, preferably n is an integer from 0 to 3, more preferably n is an integer from 0 to 2, and most preferably n is an integer from 0 to 1. Thus, the mandelic acid portion of the ester may be unsubstituted or contain one substituent (R 2 ) is most preferred.

[0034] Most preferably, R 1 and R 2 The total number of groups (ie, n+z) does not exceed six.

[0035] Substituent (R 1 ) types The inventors have found that the aromatic ring of the phenylpropanoic acid portion of the ester tolerates fluorine (-F), hydroxy (-OH) and / or alkoxy (-O-alkyl) substituents. It is understood that these substituents can also be replaced with bioisosteres known in the art. Thus, in one embodiment, R 1 (when present) are independently selected from F, OH and C1-C6 alkoxy. 1 (when present) is independently selected from OH and C1-C5 alkoxy. 1 (when present) is independently selected from OH and C1-C4 alkoxy. 1 (when present) is independently selected from OH and C1-C3 alkoxy. 1 (when present) is independently selected from OH and C1-C2 alkoxy. 1 (when present) is independently selected from OH and methoxy (OMe). In a most preferred embodiment, R 1 (when present) are independently selected from OH and C1-C3 alkoxy.

[0036] Substituent (R 2 ) types The inventors have found that the aromatic ring of the mandelic acid moiety tolerates fluorine (-F), hydroxy (-OH) and / or alkoxy (-O-alkyl) substituents. It is understood that these substituents can also be replaced with bioisosteres known in the art. Thus, in one embodiment, R 2 (when present) are independently selected from F, OH and C1-C6 alkoxy. 2 (when present) is independently selected from OH and C1-C5 alkoxy. 2(when present) is independently selected from OH and C1-C4 alkoxy. 2 (when present) is independently selected from OH and C1-C3 alkoxy. 2 (when present) is independently selected from OH and C1-C2 alkoxy. 2 (when present) is independently selected from OH and methoxy (OMe). In a most preferred embodiment, R 2 (when present) are independently selected from OH and C1-C3 alkoxy.

[0037] R 1 Position of the substituent In a preferred embodiment, (R 1 ), when present, are located at the 3-, 4- and / or 5-positions (i.e., meta and / or para), as shown in formula (I) below. [ka]

[0038] In another preferred embodiment, (R 1 ), when present, are located at the 2- and / or 6-positions (i.e., ortho), as shown in formula (I) below. [ka]

[0039] R 2 Position of the substituent In a preferred embodiment, R 2 When present, the group(s) are located at the 3-, 4- and / or 5-positions (i.e., meta and / or para), as shown in formula (I) below. Most preferably, R 2 is in fourth place. [ka]

[0040] R 3 substituent In one embodiment, R 3 is selected as H or C1-C7 alkyl, more preferably R 3 is selected as H or C1-C6 alkyl. In a preferred embodiment, R 3 is selected as H or C1-C5 alkyl. In a more preferred embodiment, R 3 is selected as H or C1-C4 alkyl. In a highly preferred embodiment, R 3 is selected as H or C1-C3 alkyl. In a more highly preferred embodiment, R 3 is selected as H or C1-C2 alkyl. In an even more highly preferred embodiment, R 3 is selected as H or C alkyl (methyl). In the most preferred embodiment, R 3 is chosen as H.

[0041] Preferred Embodiments In one embodiment, z is an integer of 0, 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, 2, or 3; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0042] In another embodiment, z is an integer of 0, 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, 2, or 3; R 2 (when present) is independently selected from OH and C1-C2 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0043] In another embodiment, z is an integer of 0, 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, 2, or 3; R 2 (when present) is independently selected from OH and methoxy (OMe); R 3 is selected as H or C1-C3 alkyl.

[0044] In another embodiment, z is an integer of 0, 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C2 alkoxy; n is an integer of 0, 1, 2, or 3; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0045] In another embodiment, z is an integer of 0, 1, 2, or 3; R 1 (when present) is independently selected from OH and methoxy (OMe); n is an integer of 0, 1, 2, or 3; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0046] In one embodiment, z is an integer of 0, 1, or 2; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0047] In another embodiment, z is an integer of 0, 1, or 2; R 1(when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C2 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0048] In another embodiment, z is an integer of 0, 1, or 2; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and methoxy (OMe); R 3 is selected as H or C1-C3 alkyl.

[0049] In another embodiment, z is an integer of 0, 1, or 2; R 1 (when present) is independently selected from OH and C1-C2 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0050] In another embodiment, z is an integer of 0, 1, or 2; R 1 (when present) is independently selected from OH and methoxy (OMe); n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0051] In another embodiment, z is an integer of 0, 1, or 2; R 1 (when present) is independently selected from OH and methoxy (OMe); n is an integer of 0, 1, or 2; R 2(when present) is independently selected from OH and methoxy (OMe); R 3 is selected as H or C1-C3 alkyl.

[0052] In a highly preferred embodiment, z is an integer of 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 3 is selected as H or C1-C3 alkyl.

[0053] In a highly preferred embodiment, z is an integer of 0, 1, 2, 3, 4, or 5, and R 1 is independently selected from F, OH, and C1-C3 alkoxy; n is an integer of 0, 1, 2, 3, 4, or 5, and R 2 is independently selected from F, OH, and C1-C3 alkoxy; R 3 is selected from H and C1-C8 alkyl; 1 or R 2 At least one of them is OH.

[0054] In another highly preferred embodiment, z is an integer of 1, 2, or 3; R 1 (when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 2 is present in the 3rd, 4th and / or 5th positions (i.e., meta or para); R 3 is selected as H or C1-C3 alkyl.

[0055] In another highly preferred embodiment, z is an integer of 1, 2, or 3; R 1(when present) is independently selected from OH and C1-C3 alkoxy; n is an integer of 0, 1, or 2; R 2 (when present) is independently selected from OH and C1-C3 alkoxy; R 2 is present at positions 2 and / or 6 (i.e., ortho); R 3 is selected as H or C1-C3 alkyl.

[0056] In a highly preferred embodiment, the compound of formula (I) is mandelic acid 2-hydroxy-phenylpropanoate, mandelic acid 3-hydroxy-phenylpropanoate, mandelic acid 4-hydroxy-phenylpropanoate, mandelic acid 3,4-dihydroxyphenylpropanoate, mandelic acid 3,4,5-trihydroxyphenylpropanoate, mandelic acid 2-methoxyphenylpropanoate, mandelic acid 3-methoxyphenylpropanoate, mandelic acid 4-methoxyphenylpropanoate, mandelic acid 4-hydroxyphenylpropanoate, Mandelic acid 4-hydroxy-3,5-dimethoxyphenylpropanoate, Mandelic acid 2,3-dimethoxyphenylpropanoate, Mandelic acid 2,4-dimethoxyphenylpropanoate, Mandelic acid 2,5-dimethoxyphenylpropanoate, Mandelic acid 3,4-dimethoxyphenylpropanoate, Mandelic acid 3,5-dimethoxyphenylpropanoate, Mandelic acid 2,3,4-trimethoxyphenylpropanoate, Mandelic acid 3,4,5-trimethoxyphenylpropanoate Mandelic acid 4-ethoxyphenylpropanoate, Mandelic acid 4-propoxyphenylpropanoate, Mandelic acid 2-hydroxy-phenylpropanoate, Mandelic acid 3-hydroxy-phenylpropanoate, Mandelic acid 4-hydroxy-phenylpropanoate, Mandelic acid 3,4-dihydroxyphenylpropanoate, Mandelic acid 2-methoxyphenylpropanoate, Mandelic acid 3-methoxyphenylpropanoate, Mandelic acid 4-methoxyphenylpropanoate, Mandelic acid 4 -Hydroxy-3-methoxyphenylpropanoate, Mandelic acid 3,5-dimethoxyphenylpropanoate, Mandelic acid 3,4,5-trimethoxyphenylpropanoate, Mandelic acid 4-ethoxy-phenylpropanoate, Mandelic acid 4-propoxy-phenylpropanoate, 4-hydroxymandelic acid 2-hydroxyphenylpropanoate, 4-hydroxymandelic acid 3-hydroxyphenylpropanoate, 4-hydroxymandelic acid 4-hydroxyphenylpropanoate, 4-hydroxymandelic acid 3,4-dihydroxyphenyl-propanoate, 4-hydroxymandelic acid 2-methoxyphenylpropanoate, 4-hydroxymandelic acid 3-methoxyphenylpropanoate, 4-hydroxymandelic acid 4-methoxyphenylpropanoate, 4-hydroxymandelic acid 4-hydroxy-3-methoxyphenyl-propanoate, 4-hydroxymandelic acid 3,5-dimethoxyphenyl-propanoate, 4-hydroxymandelic acid 4-ethoxyphenylpropanoate, 4-hydroxymandelic acid 4-propoxyphenylpropanoate, 4-methoxymandelic acid 2-hydroxy-phenylpropanoate, 4-methoxymandelic acid 3-hydroxy-phenylpropanoate, 4-methoxymandelic acid 4-hydroxyphenyl-propanoate, 4-methoxymandelic acid 3,4-dihydroxyphenylpropanoate, 4-methoxymandelic acid 2-methoxyphenylpropanoate, 4-methoxymandelic acid 3-methoxyphenylpropanoate, 4-Methoxymandelic acid 4-methoxyphenylpropanoate, 4-methoxymandelic acid 4-hydroxy-3-methoxyphenylpropanoate, 4-methoxymandelic acid 3,5-dimethoxyphenylpropanoate, 4-methoxymandelic acid 4-ethoxyphenylpropanoate, 4-methoxymandelic acid 4-propoxyphenylpropanoate, 4-propoxymandelic acid 2-hydroxyphenylpropanoate, 4-propoxymandelic acid 3-hydroxyphenylpropanoate Panoate, 4-propoxymandelic acid 4-hydroxyphenyl-propanoate, 4-propoxymandelic acid 3,4-dihydroxyphenylpropanoate, 4-propoxymandelic acid 2-methoxyphenylpropanoate, 4-propoxymandelic acid 3-methoxyphenylpropanoate, 4-propoxymandelic acid 4-methoxyphenylpropanoate, 4-propoxymandelic acid 4-hydroxy-3-methoxyphenyl-propanoate, 4-propoxymandelic acid 3,5-Dimethoxyphenyl-propanoate, 4-Propoxymandelic acid 4-ethoxyphenylpropanoate, 4-Propoxymandelic acid 4-propoxyphenylpropanoate, 4-Hydroxy-3-methoxymandelic acid 2-hydroxyphenylpropanoate, 4-Hydroxy-3-methoxy-mandelic acid 3-hydroxy-phenylpropanoate, 4-Hydroxy-3-methoxy-mandelic acid 4-hydroxy-phenylpropanoate, 4-Hydroxy-3-methoxy-mandelic acid 3,4-dihydro 4-hydroxy-3-methoxy-mandelic acid 2-methoxy-phenylpropanoate, 4-hydroxy-3-methoxy-mandelic acid 3-methoxy-phenylpropanoate, 4-hydroxy-3-methoxy-mandelic acid 4-methoxy-phenylpropanoate, 4-hydroxy-3-methoxy-mandelic acid 4-hydroxy-3-methoxy-phenylpropanoate, 4-hydroxy-3-methoxy-mandelic acid 3,5-dimethoxy-phenylpropanoate, 4-hydroxy mandelate 4-hydroxy-3-methoxy-phenylpropanoate, 4-hydroxy-3-methoxy-mandelic acid 4-ethoxy-phenylpropanoate, and 4-hydroxy-3-methoxy-mandelic acid 4-propoxy-phenylpropanoate, methyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, methyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, ethyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, ethyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, isopropyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, isoamyl mandelate 4-hydroxy-3-methoxy-phenylpropanoate, benzyl mandelate 3,4-dihydroxyphenylpropanoate, phenethyl mandelate 3,4-dihydroxyphenylpropanoate, wherein the compound may be the (S) or (R) enantiomer or any mixture thereof. ,

[0057] In a more highly preferred embodiment, the compound is mandelic acid 3-hydroxyphenylpropanoate, mandelic acid 4-hydroxyphenylpropanoate, mandelic acid 3,4-dihydroxyphenylpropanoate mandelic, mandelic acid 4-hydroxy-3-methoxyphenylpropanoate, mandelic acid 2-methoxyphenylpropanoate, mandelic acid 3-methoxyphenylpropanoate, mandelic acid 4-methoxyphenylpropanoate, mandelic acid 4-ethoxyphenylpropanoate, mandelic acid 4-propoxyphenylpropanoate, mandelic acid Mandelic acid 2,5-dimethoxyphenylpropanoate, mandelic acid 3,5-dimethoxyphenylpropanoate, mandelic acid 3,4,5-trimethoxyphenylpropanoate, 4-hydroxymandelic acid 2-methoxyphenylpropanoate, 4-hydroxymandelic acid 3-hydroxyphenylpropanoate, 4-hydroxymandelic acid 4-hydroxyphenylpropanoate, 4-hydroxymandelic acid 4-hydroxy-3-methoxyphenylpropanoate, 4-hydroxymandelic acid 3,5-dimethoxy-phenylpropanoate, 4-methoxy Mandelic acid 2-methoxyphenylpropanoate, 4-methoxymandelic acid 4-methoxyphenylpropanoate, 4-methoxymandelic acid 3,5-dimethoxyphenylpropanoate, 4-methoxymandelic acid 4-ethoxyphenylpropanoate, 4-propoxymandelic acid 4-hydroxyphenylpropanoate, 4-propoxymandelic acid 2-methoxyphenylpropanoate, 4-propoxymandelic acid 3-methoxyphenylpropanoate, 4-propoxymandelic acid 3,5-dimethoxyphenylpropanoate, 4-propoxymandelic acid acid 4-hydroxy-3-methoxyphenylpropanoate, 4-propoxymandelic acid 4-ethoxyphenylpropanoate, 4-propoxymandelic acid 4-propoxyphenylpropanoate, 4-hydroxy-3-methoxymandelic acid 4-methoxyphenylpropanoate, 4-hydroxy-3-methoxymandelic acid 3,5-dimethoxyphenylpropanoate and 4-hydroxy-3-methoxy-mandelic acid 4-ethoxyphenylpropanoate, wherein the compounds may be the (S) or (R) enantiomers or any mixture thereof.

[0058] In the most preferred embodiments, the compound is 2-((3-(4-hydroxyphenyl)propanoyl)oxy)-2-phenylacetic acid (i.e., mandelic acid 4-hydroxyphenylpropanoate), 2-((3-(3,4-dihydroxyphenyl)propanoyl)oxy)-2-phenylacetic acid (i.e., mandelic acid 3,4-dihydroxyphenyl-propanoate), or 2-((3-(4-hydroxy-3-methoxyphenyl)propanoyl)oxy)-2-phenylacetic acid (i.e., mandelic acid 4-hydroxy-3-methoxyphenylpropanoate).

[0059] Medical Use The compounds of the present invention can be used to treat diseases or disorders associated with increased levels of cellular senescence, particularly skin diseases, rheumatic diseases and / or wound healing.The compounds of the present invention can also be used to treat cancer, particularly skin cancer.

[0060] Thus, in a second aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament. In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use as a medicament in the treatment of skin diseases, rheumatic diseases, skin cancer and / or wounds.

[0061] Thus, the present invention also relates to a method of treating skin diseases, rheumatic diseases, skin cancer and / or wounds, the method comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0062] In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a skin disease or disorder selected from the group consisting of asteatotic eczema, stasis dermatitis, lichen simplex chronicus, seborrheic dermatitis, seborrhea, psoriasis, atopic dermatitis, infantile eczema, childhood eczema, adult eczema, keratosis pilaris, ichthyosis vulgaris, hand and foot dermatitis, keratoconus, dyshidrotic eczema, discoid eczema, nummular eczema, allergic contact dermatitis, irritant contact dermatitis, overtreatment dermatitis, hand eczema, and sun damage.

[0063] In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a rheumatic disease or disorder selected from the group consisting of rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, Reiter's syndrome, psoriatic arthritis, gout, juvenile chronic arthritis, enteropathic synovitis, infectious arthritis, soft tissue rheumatism and fibromyalgia.

[0064] In one embodiment, the present invention provides a method for treating acute or chronic skin wounds; acute or chronic wounds associated with bodily tissues selected from muscle, fat, bone, internal organs, nervous tissue, cartilage, joints, arteries, veins, gastrointestinal tract, mucous membranes and eyes; acute wounds selected from traumatic wounds, surgical wounds, infected wounds, mucosal wounds, burn wounds, wounds resulting from underlying medical conditions and corneal ulcers; surgical wounds, traumatic wounds, burn wounds, infected or contaminated wounds, venous ulcers, arterial ulcers, mixed venous and arterial ulcers. The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in treating a wound selected from among ulcers, pressure ulcers, diabetic ulcers, neuropathic ulcers, fistulas, immunological ulcers, malignant ulcers, dermatitis ulcers, radiation ulcers, pyoderma gangrenosum and skin graft wounds; traumatic wounds selected from among cuts, crushed wounds, ruptures, lacerations, contusions, abrasions and abrasions; and wounds that heal poorly and / or slowly.

[0065] In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a skin cancer selected from the group consisting of basal cell carcinoma, squamous cell carcinoma, melanoma, Merkel cell carcinoma, cutaneous T-cell lymphoma, dermatofibrosarcoma protuberans, Merkel cell carcinoma, and sebaceous gland carcinoma.

[0066] In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a precancerous condition, such as actinic keratosis.

[0067] Pharmaceutical Composition The compound of the present invention can be in the form of pharmaceutical composition.The pharmaceutical composition can contain pharmaceutically acceptable carriers and / or one or more excipients that are commonly used in the art.The pharmaceutical composition can be formulated as, for example, emulsion, liniment, solution, gel, foam, tablet, capsule, powder, etc.

[0068] Cosmetic or nutritional compositions and their uses The compounds of the present invention can also be used in cosmetics or nutritional compositions to prevent or reduce signs of aging in healthy subjects.Cosmetics or nutritional compositions can be used to reduce or counteract wrinkles / fine lines, age spots, hyperpigmentation and / or solar lentigines.Cosmetics or nutritional compositions can also be used to increase skin smoothness, dermal density, skin microcirculation, and / or skin moisturization.

[0069] Therefore, in a third aspect, the present invention relates to the use of the compound of formula (I) or its pharmaceutically acceptable salt in reducing or counteracting wrinkles / fine lines, age spots, hyperpigmentation and / or solar lentigines, or increasing skin smoothness, dermal density, skin microcirculation and / or skin moisturizing.Therefore, the present invention also relates to a method for reducing or counteracting wrinkles / fine lines, age spots, hyperpigmentation and / or solar lentigines, or increasing skin smoothness, dermal density, skin microcirculation and / or skin moisturizing, comprising administering an effective amount of the compound of formula (I) or its pharmaceutically acceptable salt to a subject.The compound can be administered, for example, by applying a cosmetic composition to at least a part of the skin, or by ingesting / consuming a nutritional composition.

[0070] In this context, the term "cosmetic composition" refers to a cosmetic or skin care product containing the esters of the present invention added to provide a non-therapeutic benefit to healthy subjects. Non-limiting examples include products commonly referred to as "dermocosmetics" or "anti-aging skin care," e.g., products designed to counteract the signs of aging or the aging process itself in healthy subjects. Regulatory definitions and names of such products vary in various regions of the world and change regularly. Non-limiting examples of such products include creams, lotions, gels, serums, liniments, foams, pastes, sprays, serums, solutions, powders, shampoos, and conditioners. In this context, the term "nutritional composition" refers to a food or non-food product containing the esters of the present invention added to provide a non-therapeutic physiological benefit. Non-limiting examples include products commonly referred to as functional foods, food supplements, health supplements, dietary supplements, nutraceuticals, or medical foods. Regulatory definitions and names of such products vary widely in various regions of the world and change regularly. Nutritional compositions in the form of "foods" may be in the form of specialized food preparations, including soft drinks, juices, smoothies, dairy products, etc., or general foods or beverages. Nutritional products in "non-food" form can be in the form of, for example, tablets, capsules, powders, chewing gum and lozenges. [Example]

[0071] Example 1

[0072] screening (RS)-Mandelic acid 3,4-dihydroxyphenylpropanoate was identified as a modulator of the pharmacological targets histone acetyltransferase p300 (p300), phosphodiesterase 4 (PDE4), and Kelch-like ECH-associated protein 1 (KEAP1) / nuclear factor erythroid 2-related factor 2 (NRF2). Analogs of (RS)-Mandelic acid 3,4-dihydroxyphenylpropanoate (i.e., the compound of Formula (I)) were identified by applying artificial intelligence to screen thousands of analogs for potential effects on molecular targets. Analysis was performed using a combination of computational tools, including Schrodinger's Small Molecule Suite (shape screening, pharmacophore screening, and docking analysis) and ADMET Predictor from Simulations Plus.

[0073] Exemplary compounds of Formula (I) were then synthesized and tested for their ability to modulate such targets (see Example 2 for data) and counteract the aging phenotype in three models of dermal aging reflecting intrinsic and extrinsic induction of aging (see Example 3 for data). Additionally, the ability to enhance DNA repair was demonstrated in two models of UV-induced cyclobutane pyrimidine dimer DNA damage that are therapeutically relevant to aging as well as skin cancer.

[0074] Materials and Chemicals All chemicals used were of standard analytical or synthetic grade. (R)-Mandelic acid (product number 154210), (S)-Mandelic acid (product number 778052), isoamyl (RS)-mandelate (product number S351512), phenethyl (RS)-mandelate (product number S679380), benzyl (RS)-mandelate (product number S679399), and oxalyl chloride (product number 221015) were purchased from Merck (Germany). (RS)-4-Hydroxymandelic acid (product number TCIAH0660) was purchased from Avantor (Denmark). (Berlin, Germany). (RS)-4-Methoxymandelic acid (product number BD98560), (RS)-4-propoxymandelic acid (product number BD22647), (RS)-4-hydroxy-3-methoxymandelic acid (product number BD6743), methyl (R)-mandelate (product number BD6677), methyl (S)-mandelate (product number BD32621), ethyl (R)-mandelate (product number BD127771), and ethyl (S)-mandelate (product number BD135652) were purchased from BLD Pharmatech (Germany). All hydroxyphenylpropanoic acids were provided by Zentexia ApS (Denmark). Solid-phase extraction was performed using Discovery® DSC-18 SPE Tubes (product number 52609-U) purchased from Merck (Denmark).

[0075] Synthesis of the Compounds (Esters) of the Present Invention Exemplary esters of the present invention were prepared according to the following procedure: 0.4 mmol of phenylpropanoic acid (derivative) was dissolved in a mixture of methylene chloride and tetrahydrofuran. 0.45 mmol of oxalyl chloride was added under argon, and the mixture was allowed to react at ambient temperature for 5-10 minutes to yield the acid chloride. 0.4 mmol of mandelic acid (derivative) was dissolved in tetrahydrofuran and added to the acid chloride solution under argon, and the mixture was allowed to react at ambient temperature for 45 minutes to yield the ester of the present invention. The ester solution was dried on a rotary evaporator and dissolved in a mixture of ethyl acetate and methylene chloride. The solution was transferred to a separatory funnel and washed four times with brine. The organic phase was dried over anhydrous sodium sulfate and then dried on a rotary evaporator to yield the ester of the present invention. Optionally, further purification was performed using standard solid-phase extraction procedures, where the crude ester solution was loaded onto a Discovery® DSC-18 SPE Tube and eluted with gradually increasing amounts of ethanol, from 100% water to 99.5% ethanol. The purity of the esters of the present invention is assessed by HPLC and LCMS using an Agilent 1200 HPLC equipped with a DAD detector and a 4610 QQQ detector (ESI negative mode). Elution was carried out on an Agilent Technologies Poroshell 120 SB-C18 column (3 × 150 mm, 2.7 μm) with a gradient of water and acetonitrile (both containing 0.1% formic acid).

[0076] The following esters of the invention were prepared: [Table 1] TIFF2025537789000006.tif241147TIFF2025537789000007.tif253150TIFF20255377890 00008.tif245145TIFF2025537789000009.tif193112TIFF2025537789000010.tif251146 TIFF2025537789000011.tif231146TIFF2025537789000012.tif192109TIFF20255377890 00013.tif193110TIFF2025537789000014.tif192110TIFF2025537789000015.tif168145

[0077] conclusion The compound of formula (I) was successfully synthesized and purified by the procedures described.

[0078] Example 2

[0079] the purpose The aim was to test the modulatory effect of compounds of formula (I) on three pharmacological targets related to cellular senescence.

[0080] Test Compound The test compounds prepared in Example 1 were dissolved in DMSO prior to testing.

[0081] Phosphodiesterase 4D (PDE4D) PDE4D hydrolyzes the second messenger cAMP, a regulator and mediator of several cellular responses to extracellular signals. PDE4D plays a particularly important role in skin aging, and elevating cAMP through inhibition of PDE4D holds the potential to alleviate the aging-associated secretory phenotype (SASP) of the skin.

[0082] Assay The PDE-Glo™ Phosphodiesterase Assay (Catalog No. V1361) was purchased from Promega (USA). Active PDE4D (Catalog No. P92-31DG-05) was purchased from SignalChem Biotech (Canada). PDE4D stock (0.1 μg / mL) was diluted in assay buffer. The assay was performed in a 96-well format according to the manufacturer's protocol. All dilutions were performed in assay buffer. Rolipram was used as a positive inhibitor control. The procedure was as follows: 10 μl of diluted PDE4D was added to each well, followed by 5 μL of test compound solution or vehicle control. The reaction was initiated by adding 10 μL of 2.5 μM cAMP to each well, followed by incubation at room temperature for 1 hour. The reaction was stopped by adding 12.5 μL of PDE-Glo™ Termination Buffer to each well, followed by 12.5 μL of PDE-Glo™ Detection Solution. Plates were incubated at room temperature for 20 minutes, after which 50 μL of luciferase-based Kinase-Glo™ Reagent was added to each well, followed by a 10-minute incubation at room temperature. Chemiluminescence was measured on a Spectramax ID5 (Molecular Devices) with an integration time of 1000 ms.

[0083] result After subtracting the background control, the concentration that inhibited the assay by 50% (IC-50) compared to the vehicle control was established for each test compound (see table below).

[0084] [Table 2] TIFF2025537789000017.tif230132TIFF2025537789000018.tif25130

[0085] conclusion Test compounds of the invention showed convincing inhibition of PDE4D, supporting more general findings of efficacy in AI-based (computer) models of compounds of formula (I) as described in Example 1.

[0086] Histone acetyltransferase p300 (p300) Histone acetyltransferase (HAT) enzymes coordinate the acetylation of histone and non-histone proteins. p300 is a transcriptional coactivator that acetylates core histones to promote chromatin decondensation and recruits the basal RNA polymerase machinery. In addition, many non-histone proteins, such as p53, STATs, and alpha interferon receptors, serve as substrates for p300.

[0087] Assay The P300 Chemiluminescent Assay Kit (Cat. No. 79705) was purchased from BSP Bioscience Inc. (USA). The assay was performed in a 96-well format according to the manufacturer's protocol. All dilutions were performed in assay buffer. Anacardic acid was used as a positive inhibitor control. The procedure was as follows: 5 μL of test compound solution or vehicle control was added to a 96-well plate precoated with histone peptides. 20 μL of p300 solution (0.01 ng / μL) was added to all wells except the blank and negative control, which received 20 μL assay buffer. The plate was incubated at room temperature for 30 minutes. 25 μL of acetyl-CoA substrate solution was added to each well, followed by incubation at 30°C for 1 hour. The supernatant was removed, and the plate was washed three times with 200 μL of TBST buffer. 100 μL of blocking buffer was then added (left for 10 minutes), and the plate was washed three times with 200 μL of TBST buffer. 100 μL of diluted primary antibody was added to each well, followed by incubation at room temperature for 1 hour. The antibody is specific for the acetylated form of the histone peptide, and therefore, binding was proportional to p300 activity.

[0088] The supernatant was removed, and the plate was washed three times with 200 μL of TBST buffer. 100 μL of blocking buffer was added (left for 10 minutes), followed by three additional washes with 200 μL of TBST buffer. 100 μL of diluted secondary HRP-labeled antibody was added to each well, followed by incubation at room temperature for 1 hour. The supernatant was removed, and the plate was washed three times with 200 μL of TBST buffer. 100 μL of blocking buffer was added (left for 10 minutes), followed by three additional washes with 200 μL of TBST buffer.

[0089] 100 μL of a mixture of ELISA ECL substrates A and B was added to each well, and chemiluminescence was read on a Spectramax ID5 (Molecular Devices) with an integration time of 1000 ms.

[0090] result The concentration that inhibited the assay by 50% (IC-50) compared to vehicle control was established for each test compound (see table below).

[0091] [Table 3]

[0092] conclusion The test compounds showed convincing inhibition of p300, confirming the more general findings of efficacy in AI-based (computer) models of compounds of formula (I) as described in Example 1.

[0093] Kelch-like ECH-associated protein 1 (KEAP1) / nuclear factor erythroid 2-related factor 2 (NRF2) The NRF2 antioxidant response pathway plays an important role in cellular defense and regeneration. NRF2, a basic leucine zipper transcription factor, induces the expression of antioxidant and phase II enzymes by binding to the antioxidant response element (ARE) region of gene promoters. Under basal conditions, NRF2 is retained in the cytosol by binding to the cytoskeletal protein KEAP1. Upon exposure to oxidative stress or other ARE activators, NRF2 is released from KEAP1 and translocates to the nucleus, where it can bind to AREs, leading to the expression of antioxidant and phase II enzymes that protect cells from oxidative damage. Therapeutic agents that release NRF2 by binding to KEAP1 can activate the antioxidant response pathway without oxidative stress.

[0094] Assay The ARE Reporter-Hep G2 cell line (catalog no. 60513), ONE-Step™ Luciferase Assay System (catalog no. 60690), and Growth Medium 1K (catalog no. 79533) were purchased from BSP Bioscience (USA). Cells were cultured in Growth Medium 1K according to the manufacturer's protocol until the day of assay. The assay was performed in a 96-well format according to the manufacturer's protocol. All dilutions were performed in Growth Medium 1K. 80 μM methyl fumarate was used as a positive control. The procedure was as follows: Hep G2 cells were seeded at a density of 40,000 cells / well in 45 μL of assay medium into a white, clear-bottom 96-well microplate. 10 μL of test compound solution or vehicle control was added to each well, followed by incubation at 37°C and 5% CO2 for 18 hours. 100 μL of ONE-Step™ Luciferase Assay Reagent was added to each well, followed by shaking at room temperature for 15 minutes. ARE luciferase reporter transcriptional responses were measured as chemiluminescence on a Spectramax ID5 (Molecular Devices), and relative ARE expression was calculated after subtraction of background controls.

[0095] result Relative ARE expression was calculated after subtracting background controls. The concentration inducing a 50% enhanced ARE transcriptional response (EC-50) was established for each test compound (see table below).

[0096] [Table 4]

[0097] conclusion The test compounds showed convincing activation of the NRF2 pathway, confirming more general findings of efficacy in AI-based (computer) models of compounds of formula (I) as described in Example 1.

[0098] Example 3

[0099] the purpose To establish whether (R)-mandelic acid 3,4-dihydroxyphenylpropanoate can counteract the aging phenotype, seven studies of intrinsic or extrinsic dermal aging were performed.

[0100] Test Compound (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate was prepared according to Example 1.

[0101] method A total of seven studies (Hernandez-Segura et al., 2018) in three models of dermal aging reflecting intrinsic and extrinsic induction of aging were used:

[0102] 1. Intrinsic aging in primary human dermal fibroblasts. -In this endogenous model, the aging phenotype was mediated by replicative telomere shortening. -Cells were cultured until they stopped proliferating (typically >20 passages), after which they acquired a senescent phenotype. These cells were seeded into 6-well plates. Non-senescent cells (<10 passages) from the same batch were seeded as a non-senescent control. -Senescent cells were incubated for 48 hours with a concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate or vehicle, as well as vehicle-treated non-senescent (normal) controls. -This was followed by a 3 hour incubation with resazurin and relative cell density and proliferation was established by measuring the fluorescence (excitation 550 nm and emission 590 nm) of the collected medium. -Cells were harvested and gene expression was analyzed by real-time qRT-PCR.

[0103] 2. DNA damage-induced aging in human dermal fibroblasts (fHDF / TERT166). In this exogenous model, the senescent phenotype was induced with doxorubicin. Cells were treated with doxorubicin for 24 hours, followed by incubation without doxorubicin for 6 days, after which the cells acquired a senescent phenotype. These cells were seeded into 6-well plates. Cells from the same batch that were not treated with doxorubicin were seeded as non-senescent controls. -Senescent cells were incubated for 48 hours with a concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate or vehicle, as well as vehicle-treated non-senescent (normal) controls. -This was followed by a 3 hour incubation with resazurin and relative cell density and proliferation was established by measuring the fluorescence (excitation 550 nm and emission 590 nm) of the collected medium. -Cells were harvested and gene expression was analyzed by real-time qRT-PCR.

[0104] 3. Oxidative stress-induced aging in human dermal fibroblasts (fHDF / TERT166). In this exogenous model, the senescence phenotype was induced with H2O2. The cells were transiently treated with H2O2 for 6 days, after which the cells acquired a senescent phenotype. These cells were seeded into 6-well plates. The same batch of cells that were not treated with H2O2 were seeded as a non-senescent control. -Senescent cells were incubated for 48 hours with a concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate or vehicle, as well as vehicle-treated non-senescent (normal) controls. -This was followed by a 3 hour incubation with resazurin and relative cell density and proliferation was established by measuring the fluorescence (excitation 550 nm and emission 590 nm) of the collected medium. -Cells were harvested and gene expression was analyzed by real-time qRT-PCR.

[0105] Key Comparisons In each model, the expression of key genes involved in skin aging was measured. All models included: Normal control (non-aging, vehicle-treated) Aged control (vehicle-treated) and · Senescent cells treated with (R)-mandelic acid 3,4-dihydroxyphenylpropanoate (in vehicle).

[0106] This design allowed for the following important comparisons: Aging controls versus normal controls to establish whether significant induction of aging-related gene expression occurred. · (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate treated cells versus senescent controls to establish whether (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate inhibited aging-related gene expression. (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate treated cells versus normal controls to establish whether (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate treatment can revert cells to a non-senescent state.

[0107] [Table 5]

[0108] [Table 6]

[0109] [Table 7]

[0110] [Table 8]

[0111] result To get the best overview of the observed effects as well as reproducibility between models, the data were examined by gene expression across the seven skin tests.

[0112] In all of the data presented below, gene expression was normalized to two reference (housekeeping) genes, GAPDH and ACTB.

[0113] Effects on dermal aging and key markers of aging CDKN1A(p21 cip1 / waf1 ) CDKN1A is a well-established marker of senescence in dermal fibroblasts and has also been found to correlate with age in biopsies from young, middle-aged or elderly humans (Idda et al., 2020).

[0114] Intrinsic Aging Research In both intrinsic aging studies, CDKN1A was significantly (p<0.01 or p<0.05) overexpressed (2.1-5.3 fold) in aged controls compared to normal controls. In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.05) reduced the expression of CDKN1A compared to aged controls, corresponding to: -0.23 to 0.50 times at 2.0 μg / ml -0.22 to 0.54 times at 8.0 μg / ml

[0115] All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups in both studies showed CDKN2A expression that was not significantly different compared to normal controls.

[0116] DNA damage-induced aging research In both DNA damage-induced aging studies, CDKN1A was significantly (p<0.05) overexpressed (3.9-4.7-fold) in aged controls compared to normal controls.

[0117] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.05) reduced CDKN1A expression compared to aged controls, corresponding to: -0.29 to 0.30 times at 2.0 μg / ml -0.26 to 0.36 times at 8.0 μg / ml

[0118] All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treated groups in both studies showed CDKN2A expression that was close (not significant) to or only slightly higher than normal controls.

[0119] Oxidative stress-induced aging research In all three oxidative stress-induced aging studies, CDKN1A expression was significantly (p<0.01 or p<0.05) enhanced (2.1-3.6-fold) in aged controls compared with normal controls.

[0120] In all studies and at all concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.05) reduced CDKN1A expression compared to aged controls, corresponding to: -0.47 times at 1.0 μg / ml -0.29 to 0.30 times at 2.0 μg / ml -0.37 times at 4.0 μg / ml - and 0.26-0.36 times at 8.0 μg / ml.

[0121] In two of the three studies, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate concentrations of 2.0 mg / ml and 8.0 μg / ml resulted in significantly (p<0.01 or p<0.05) decreased CDKN1A expression compared to normal controls.

[0122] Conclusion CDKN1A In all three aging models, CDKN1A was significantly overexpressed in aged controls, confirming the aging state. Collectively, the data convincingly demonstrate that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate, at concentrations achievable by topical administration, reduces CDKN1A expression in aged dermal fibroblasts to non-aging levels associated with younger skin, regardless of the mode of senescence induction.

[0123] LMNB1 (lamin B1) LMNB1 (lamin B1) expression is significantly downregulated in senescent dermal fibroblasts, and its decrease has also been proposed as a marker for quantifying cellular senescence in photoaged skin (Wang et al., 2017). For each study, the relative expression of LMNB1 was shown separately for senescent and normal controls.

[0124] Intrinsic Aging Research In both intrinsic aging studies, the relative expression of LMNB1 was significantly reduced (0.11- to 0.16-fold) in aged controls compared with normal controls, whereas both concentrations of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate enhanced expression to levels slightly lower than normal controls, but not significantly different.

[0125] All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate groups in both studies showed significantly (p<0.05) enhanced LMNB1 expression compared to aged controls, corresponding to: -5.0 to 8.9 times at 2.0 μg / ml -4.9 to 7.0 times at 8.0 μg / ml

[0126] DNA damage-induced aging research In both DNA damage-induced aging studies, the relative expression of LMNB1 was significantly (p<0.01) reduced (0.026-0.029-fold) in aged controls compared to normal controls. All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups in both studies showed significantly (p<0.05) enhanced LMNB1 expression compared to aged controls, corresponding to: -2.0μg / ml: 25.6 to 27.4 times -23.3 to 30.3 times at 8.0 μg / ml

[0127] Both concentrations of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate enhanced LMNB1 expression to levels close to those of normal controls, as only two of the four (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups were significantly (p<0.05) lower compared to normal controls.

[0128] Oxidative stress-induced aging research In all three oxidative stress-induced aging studies, the relative expression of LMNB1 was significantly (p>0.05) reduced (0.27-0.41-fold) in aged controls compared with normal controls, whereas all tested concentrations of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate enhanced expression to levels close to or significantly (p<0.05) higher than those in normal controls.

[0129] All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate concentrations in all three studies showed significantly (p<0.05) higher LMNB1 expression compared to aged controls, corresponding to: -1.7 times at 1.0 μg / ml -2.8 to 4.0 times at 2.0 μg / ml -2.3 times at 4.0 μg / ml -3.0 to 4.0 times at 8.0 μg / ml

[0130] Conclusion LMNB1 In all three aging models, LMNB1 was significantly downregulated in aged controls, confirming the aging state. Collectively, the data convincingly demonstrate that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treatment, at concentrations achievable by topical administration, enhances LMNB1 expression in aged dermal fibroblasts to non-aged levels associated with young skin, independent of the mode of senescence induction.

[0131] Effects on key markers of dermal connective tissue Collagen expression (COL1A1 and COL3A1) Collagen constitutes approximately 70–80% of the dry weight of the dermis, consisting of approximately 80% type I collagen and 15% type III collagen (Waller and Maibach 2006).

[0132] Decreased collagen expression plays a central role in the structural changes associated with skin aging and is associated with fibroblast senescence (Ezure et al., 2019). This is reflected in lower expression of type I collagen alpha 1 chain (COL1A1) and type III collagen alpha 1 chain (COL3A1).

[0133] For each study, the relative expression of COL1A1 and COL3A1 was shown separately for aged and normal controls, respectively.

[0134] Intrinsic aging test (COL1A1 only) In both intrinsic aging studies, COL1A1 was significantly (p<0.01) underexpressed (0.16-0.36-fold) compared to normal controls.

[0135] In both studies, both concentrations of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) increased COL1A1 expression compared to aged controls, corresponding to: -3.1 times at 2.0 μg / ml -7.1 times at 8.0 μg / ml

[0136] In both studies, the highest concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed a significant (p<0.05) enhancement of COL1A1 expression compared to normal controls.

[0137] DNA damage-induced aging research In both DNA damage-induced aging studies, COL3A1 was significantly (p<0.01 or p<0.05) underexpressed (0.17-0.29-fold) in aged controls compared to normal controls, whereas COL1A1 was significantly (p<0.01) underexpressed (0.40-fold) in only one study.

[0138] In both studies and at both concentrations, the (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treated groups showed significantly (p<0.05) enhanced expression of COL1A1 compared to aged controls, which corresponds to: -4.3 to 9.3 times at 2.0 μg / ml -8.0μg / ml: 5.0 to 25.3 times

[0139] Furthermore, both concentrations in both studies showed significantly (p<0.05) enhanced COL1A1 expression compared to normal controls, which corresponds to: -1.7 to 8.5 times at 2.0 μg / ml -2.0 to 23.2 times at 8.0 μg / ml

[0140] In both studies and at both concentrations, the (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treated groups showed significantly (p<0.05) enhanced expression of COL3A1 compared to aged controls, which corresponds to: -2.8 to 8.5 times at 2.0 μg / ml -3.7 to 8.5 times at 8.0 μg / ml

[0141] Three of the four (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups in the two studies showed a significant (p<0.05) increase in COL3A1 expression compared with normal controls.

[0142] Oxidative stress-induced aging In all oxidative stress-induced aging studies, COL1A1 was significantly (p<0.05) underexpressed (0.14- to 0.52-fold) in aged controls compared to normal controls.

[0143] In all studies and at all concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treated groups showed significantly (p<0.05) enhanced COL1A1 expression compared to aged controls, corresponding to: -4.1 to 5.7 times at 2.0 μg / ml -2.8 to 13.3 times at 8.0 μg / ml

[0144] Six of the eight (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups in the three studies showed a significant (p<0.05) increase in COL1A1 expression compared with normal controls.

[0145] In all oxidative stress-induced aging studies, COL3A1 was significantly (p<0.05) underexpressed (0.17-0.42-fold) in aged controls compared to normal controls.

[0146] In all studies and at all concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treated groups showed significantly (p<0.05) enhanced expression of COL3A1 compared to aged controls, which corresponds to: -4.4 to 6.2 times at 2.0 μg / ml -2.8 to 6.5 times at 8.0 μg / ml

[0147] Four of the eight (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated groups in the three studies showed significant (0<0.01 or p<0.05) enhancement of COL3A1 expression compared with normal controls.

[0148] Conclusions COL1A1 and COL3A1 In all seven studies and at all concentrations tested, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) enhanced the expression of both COL1A1 and COL3A1 compared with aged controls, reaching levels similar to, or in many cases higher than, those of normal controls. This convincingly demonstrates that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate can reverse the aging-associated downregulation of collagen expression at concentrations achievable by topical treatment.

[0149] Elastin (ELN) and TIMP metallopeptidase inhibitor 1 (TIMP1) ELNs play a major structural role in the dermis and are crucial for dermal elasticity. ELNs are significantly downregulated in skin aging (Ezure et al., 2019; Imokawa et al., 2015).

[0150] Tissue inhibitor of metalloproteinases 1 (TIMP1) exhibits potent inhibition of matrix metallopeptidases and is released by fibroblasts to control inflammatory degradation by MMPs. TIMP1 is an essential protective factor for dermal connective tissue, and its expression decreases with aging of fibroblasts both ex vivo and in vivo, thus contributing to the increased degradation of dermal connective tissue with aging (Hornebeck et al. 2003).

[0151] Intrinsic aging test (ELN only) In both intrinsic aging studies, ELN was significantly (p<0.05) underexpressed (0.24-0.38-fold) in aged controls compared to normal controls.

[0152] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) increased ELN expression compared to aged controls, corresponding to: -2.4 to 4.8 times at 2.0 μg / ml -3.5 to 3.9 times at 8.0 μg / ml

[0153] ELN expression in the (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated group did not show statistical significance compared with normal controls in any of the studies.

[0154] DNA damage-induced aging research In one study, there was a trend toward ELN underexpression (0.59-fold) in aging controls compared to normal controls, but this did not reach statistical significance (p=0.078).In another study, ELN was significantly (p<0.01) underexpressed (0.41-fold) in aging controls compared to normal controls.

[0155] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly (p<0.01 or p<0.05) enhanced ELN expression compared to aged controls, corresponding to: -2.0μg / ml: 5.4 to 13.4 times -4.5 to 32.1 times at 8.0 μg / ml

[0156] Furthermore, in both studies, all (R)-mandelic acid 3,4-dihydroxyphenylpropanoate groups showed a significant (p<0.05) enhancement of ELN expression compared to normal controls.

[0157] In one study, TIMP1 was significantly (p<0.01) underexpressed (0.87-fold) in aging controls compared to normal controls, whereas expression in the other study was not significantly different.

[0158] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) increased TIMP1 expression compared to aged controls, corresponding to: -1.6 to 4.8 times at 2.0 μg / ml -1.9 to 7.3 times at 8.0 μg / ml

[0159] In both studies, all (R)-mandelic acid 3,4-dihydroxyphenylpropanoate groups showed a significant (p<0.05) enhancement of TIMP1 expression compared to normal controls.

[0160] Oxidative stress-induced aging In all oxidative stress-induced aging studies, ELN was significantly (p<0.05) underexpressed (0.24-0.55-fold) in aged controls compared to normal controls.

[0161] In all three studies and at all concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly (p<0.05) enhanced expression of ELNs compared to aged controls, corresponding to: -1.9 times at 1.0 μg / ml -3.6 to 6.1 times at 2.0 μg / ml -3.7 times at 4.0 μg / ml -8.0μg / ml: 4.6 to 8.7 times

[0162] Furthermore, in one of three studies, 2.0 and 8.0 μg / ml of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly enhanced expression of ELN compared with normal controls.

[0163] TIMP1 expression in aged controls was significantly (p<0.05) underexpressed (0.55-fold) in only one of three oxidative stress-induced aging studies.

[0164] In all studies, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 μg / ml showed significantly (p<0.05) enhanced TIMP1 expression compared with either aged controls at 2.0 or 8.0 μg / ml.

[0165] Conclusions ELN and TIMP1 In all seven studies and at all tested concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) enhanced ELN expression compared to aged controls, reaching levels similar to or significantly (p<0.05) higher than normal controls.

[0166] This convincingly demonstrates that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate can reverse aging-associated downregulation of ELN expression at concentrations achievable by topical administration.

[0167] Furthermore, in all five studies in which TIMP1 was tested, and at most concentrations tested, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly enhanced TIMP1 expression compared to aging controls (p<0.01 or p<0.05), and in four of the five studies, it reached levels significantly enhanced compared to normal controls (p<0.05). Because TIMP1 inhibits matrix metallopeptidases, this effect may contribute to the anti-aging effects of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate.

[0168] Effects on key markers of dermal SASP The senescence-associated secretory phenotype (SASP) is a characteristic feature of dermal aging and may have a detrimental effect on connective tissue architecture through enhanced expression of matrix metallopeptidases.

[0169] Matrix metallopeptidases MMP1 and MMP3 Both MMP1 and MMP3 are overexpressed in aged skin as well as in senescent fibroblasts and are involved in aging-associated dermal connective tissue degradation (Hornebeck et al., 2003).

[0170] For each study, the relative expression of MMP1 and MMP3 was shown separately for aged and normal controls, respectively.

[0171] Intrinsic aging test (measurement of MMP1 only) In both intrinsic aging studies, MMP1 expression was significantly (p>0.05) enhanced (2.2-2.6-fold) in aged controls compared with normal controls. All (R)-mandelic acid 3,4-dihydroxyphenylpropanoate concentrations in both studies showed significantly (p<0.05) suppressed MMP1 expression compared to aged controls, which corresponds to: -0.39 to 0.42 times at 2.0 μg / ml -0.40 to 0.52 times at 8.0 μg / ml

[0172] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed similar levels of MMP1 expression compared to normal controls (not statistically significant).

[0173] DNA damage-induced aging research In both DNA damage-induced aging studies, significantly (p<0.05) enhanced expression of MMP1 (3.0-3.2-fold) and MMP3 (2.3- and 4.8-fold) was observed in aged controls compared with normal controls.

[0174] In both tests and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly suppressed the expression of MMP1 compared to aged controls, which corresponds to: -0.17 to 0.49 times at 2.0 μg / ml -0.18 to 0.60 times at 8.0 μg / ml

[0175] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly suppressed the expression of MMP3 compared to aged controls, which corresponds to: -0.25 to 0.32 times at 2.0 μg / ml -0.23 to 0.32 times at 8.0 μg / ml

[0176] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate induced MMP1 and MMP3 expression close to the levels of normal controls, reaching significantly (p<0.01 or p<0.05) lower expression in one study.

[0177] Oxidative stress-induced aging research In all oxidative stress-induced aging models, significantly (p<0.01 or p<0.05) enhanced expression of MMP1 (2.7-38.3-fold) and MMP3 (4.9-8.2-fold) was observed in aged controls compared with normal controls, whereas the (R)-mandelic acid 3,4-dihydroxyphenylpropanoate-treated group showed expression levels close to those of normal controls.

[0178] With one exception, all concentrations of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate in all studies significantly (p<0.01 or p<0.05) inhibited MMP1 expression compared to aged controls, corresponding to: -0.04 times at 1.0 μg / ml -0.03 to 0.39 times at 2.0 μg / ml -0.06 times at 4.0 μg / ml -0.04 to 0.83 times at 8.0 μg / ml

[0179] In all studies and at all concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) inhibited the expression of MMP3 compared to aged controls, which corresponds to: -0.36 times at 1.0 μg / ml -0.16 to 0.46 times at 2.0 μg / ml -0.23 times at 4.0 μg / ml -0.17 to 0.18 times at 8.0 μg / ml

[0180] Conclusion Matrix metallopeptidase In conclusion, the data convincingly demonstrate that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate, at concentrations achievable by topical administration, reduces the expression of the key SASP genes MMP1 and MMP3 in senescent dermal fibroblasts, independent of the mode of aging induction.

[0181] (R)-Mandelic acid 3,4-dihydroxyphenylpropanoate treatment was able to reduce expression to levels not different from normal controls in 4 of 7 studies for MMP1 and 2 of 5 studies for MMP3.

[0182] Effects on key markers of epigenetic aging repair DNMT1 (DNA methyltransferase 1) and SIRT1 (sirtuin 1) Lower expression of DNA methyltransferase 1 (DNMT1) is characteristic of senescent dermal fibroblasts. DNMT1 is known as a "maintenance DNMT" because it preserves the original non-senescent methylation of dermal cells. DNMT1 expression is inversely correlated with chronological age in human skin (Orioli et al., 2018).

[0183] The class III histone deacetylase SIRT1 exerts control over energy metabolism, inflammation, and oxidative stress, as well as mediating cell survival, UV damage response, DNA repair, and tissue regeneration (Garcia-Peterson et al., 2017). In dermal fibroblasts, SIRT1 expression significantly decreases with age (Tigges et al., 2014; Carlomosti et al., 2017), and notably, upregulation or downregulation of SIRT1 results in delayed or accelerated fibroblast senescence, respectively (De Cabo et al., 2015).

[0184] Intrinsic Aging Research In both intrinsic aging studies, DNMT1 was significantly (p<0.01 or p<0.05) underexpressed (0.39-0.51-fold) in aging controls compared to normal controls. In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p>0.001, p<0.01, or p<0.05) increased DNMT1 expression compared to aging controls, corresponding to: -2.7 to 2.9 times at 2.0 μg / ml -2.8 to 2.9 times at 8.0 μg / ml

[0185] Furthermore, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 and 8.0 μg / ml significantly (p<0.01 or p<0.05) enhanced DNMT1 expression compared with normal controls in some studies and was not different from controls in other studies.

[0186] In both intrinsic aging studies, SIRT1 was significantly (p<0.05) underexpressed (0.53-0.64-fold) in aged controls compared to normal controls.

[0187] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) increased SIRT1 expression compared to aged controls, corresponding to: -2.8 to 3.2 times at 2.0 μg / ml -2.8 to 3.4 times at 8.0 μg / ml

[0188] Furthermore, 2.0 and 8.0 μg / ml of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly (p>0.001, p<0.01 or p<0.05) enhanced expression of DNMT1 compared to normal controls in both studies, which corresponds to: -1.7 to 1.8 times at 2.0 μg / ml -1.8 to 1.8 times at 8.0 μg / ml

[0189] DNA damage-induced aging research In both DNA damage-induced aging studies, DNMT1 was significantly (p<0.01 or p<0.05) underexpressed (0.11-0.32-fold) in aged controls compared to normal controls.

[0190] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.01 or p<0.05) increased DNMT1 expression compared to aged controls, corresponding to: -4.6 to 5.2 times at 2.0 μg / ml -8.0μg / ml: 5.0 to 5.6 times

[0191] In one study, the highest concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly (p<0.05) enhanced DNMT1 expression compared to normal controls, whereas in other studies both concentrations were significantly (p<0.01 or p<0.05) lower.

[0192] In both DNA damage-induced aging studies, SIRT1 was significantly (p<0.01 or p<0.05) underexpressed (0.20-0.54-fold) in aged controls compared to normal controls.

[0193] In both studies and at both concentrations, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate significantly (p<0.05) increased SIRT1 expression compared to aged controls, corresponding to: -3.2 to 3.5 times at 2.0 μg / ml -2.8 to 3.1 times at 8.0 μg / ml

[0194] In one study, the highest concentration of (R)-mandelic acid 3,4-dihydroxyphenylpropanoate showed significantly (p<0.05) enhanced SIRT1 expression compared to normal controls, whereas in the other study both concentrations were significantly (p<0.05) lower.

[0195] Oxidative stress-induced aging research In two of three oxidative stress-induced aging studies, DNMT1 was significantly (p<0.05) underexpressed (0.50-0.84-fold) in aged controls compared to normal controls.

[0196] In all three studies, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 μg / ml and 8.0 μg / ml significantly (p<0.05) increased DNMT1 expression compared to aged controls, corresponding to: 1.5 to 1.9 times at 2.0 μg / ml 1.4 to 2.9 times at 8.0 μg / ml

[0197] Furthermore, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 μg / ml and 8.0 μg / ml significantly (p<0.05) enhanced DNMT1 expression compared to normal controls in two of the three studies.

[0198] SIRT1 expression showed a trend toward suppression (0.58-0.89 fold) in aging controls compared with normal controls, which was statistically significant in only one study (p<0.05).

[0199] In all three studies, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 μg / ml and 8.0 μg / ml significantly (p<0.05) increased SIRT1 expression compared to aging controls, corresponding to: 2.1 to 3.5 times at 2.0 μg / ml 1.8 to 4.2 times at 8.0 μg / ml

[0200] Furthermore, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate at 2.0 μg / ml and 8.0 μg / ml showed significant (p<0.05) enhancement of SIRT1 expression compared to normal controls in all three studies.

[0201] Conclusions DNMT1 and SIRT1 In all seven studies across three dermal aging models, (R)-mandelic acid 3,4-dihydroxyphenyl-propanoate treatment significantly and substantially enhanced the expression of DNMT1 and SIRT1 compared to aged controls, reaching levels at or significantly higher than those of normal controls.

[0202] This demonstrates that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate induces epigenetic repair pathways and thus eliminates their aging-associated downregulation at concentrations achievable with topical administration.

[0203] Overall conclusions of dermal aging research All seven studies involving three dermal aging models demonstrated key elements of the aging phenotype according to the changes seen in aged human dermis: -Cyclin-dependent kinase inhibitor p21 cip1 / waf1 A significant and substantial increase in the expression of (CDKN1A)-mediated cell cycle arrest. -A significant and substantial decrease in the expression of the nuclear lamina protein lamin B1 (LMNB1), which is essential for nuclear stability. -A significant and substantial reduction in the expression of essential connective tissue components, including proteins related to collagen I (COL1A1), collagen III (COL3A1) and elastin (ELN), which are essential for maintaining the thickness, structure and elasticity of young skin. -A significant and substantial increase in the expression of SASP-related matrix metallopeptidases, including MMP1 and MMP3, which cause age-related degradation and damage of dermal connective tissue. -Significant and substantial reduction in the expression of essential epigenetic maintenance and repair genes, including the class III histone deacetylase sirtuin 1 (SIRT1) and DNA methyltransferase 1 (DNMT1).

[0204] In all seven studies, (R)-mandelic acid 3,4-dihydroxyphenylpropanoate treatment showed significant, reproducible, and substantial reversal effects on all of the above-mentioned traits of dermal aging resulting in a non-aging phenotype associated with younger skin.

[0205] Based on the data, it can be concluded that (R)-mandelic acid 3,4-dihydroxyphenylpropanoate is effective at concentrations achievable in the dermis using topical formulations.

[0206] Example 4

[0207] the purpose To establish whether compounds of the present invention can enhance the repair of DNA damage induced by ultraviolet (UV) light exposure, two studies of aging- and cancer-associated DNA damage in human skin were conducted.

[0208] Test Compound (R)-Mandelic acid 4-hydroxy-3-methoxyphenylpropanoate was prepared according to Example 1.

[0209] method DNA damage is a major driver of cellular senescence and aging. Insufficient repair of DNA damage can further lead to mutations and cancer development. The ability to repair DNA damage gradually declines with age, and this decline plays an important role in accelerated skin aging and the risk of skin cancer.

[0210] Absorption of ultraviolet (UV) light generates two major types of DNA damage: cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts, which play a central role in skin aging and the development of human skin cancer. UV-damaged DNA is usually repaired by nucleotide excision repair or base excision repair. After UV exposure, cells arrest the cell cycle for repair. Insufficient DNA repair capacity plays an essential role in skin aging and the development of aging phenotypes, hence the term "photoaging."

[0211] Two different models of UV-induced DNA damage were used, and in both models, treatment with (R)-mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate was initiated after the induction of DNA damage to determine whether (R)-mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate could accelerate DNA repair.

[0212] 1. UV-induced DNA damage in human skin explants from middle-aged donors. In this model, 11 mm circular skin biopsies from the same donor (female, 52 years old) were provided by Genoskin, France. The explants (Nativeskin Access®) were embedded in a matrix that maintained all normal skin physiology for up to 7 days and cultured in nutrient medium in 12-well plates in an incubator at 37°C and 5% CO2.

[0213] Significant DNA damage was induced by placing the explants in a UV-solar simulator (SOL500 with UV filter H2) from Dr. Honle GmbH, Germany, which provided a combination of UVA and UVB radiation comparable to natural sunlight. Three groups of four explants were treated with 100 mJ / cm 2 The explants were exposed to 1000 uV of UVB once daily for two consecutive days. A control group of four explants without UV exposure was included to estimate the increase in DNA damage caused by UV treatment. Immediately after the last UV treatment, groups of explants were treated topically on the surface of the skin with 10 μL / explant of one of the following: 4.0% (w / w) of (R)-Mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate 2.0% (w / w) of (R)-Mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate - Vehicle (both UV and non-UV control groups) Treatment was repeated once daily for four consecutive days. On day -6, two 4 mm punch biopsies were taken and used as follows: One was used to estimate cell viability using the Resazurin Deep Blue Cell Viability Kit (Cat. No. 424702) from BioLegend (USA). Biopsies were incubated in 900 μL of fresh nutrient medium with 90 μL of Resazurin Deep Blue for 4 hours, after which the medium was collected and transferred to a black fluorescent 96-well plate (Thermo Scientific 137101), and fluorescence was measured on a Spectramax ID5 at 550 nm excitation and 590 nm emission to establish cell viability. The other biopsy was subjected to tissue homogenization in a GentleMACS™ tube from Miltenyi Biotec (Germany), followed by DNA extraction using the DNeasy® Blood & Tissue Kit (Qiagen, UK) according to standard protocols. - Double-stranded DNA (dsDNA) was quantified using the Invitrogen™ Quant-iT™ dsDNA Assay Kit (Cat. No. Q33130) from Thermo Fisher Scientific (Denmark). The amount of cyclobutane pyrimidine dimer (CPD) in each biopsy was quantified with a specific antibody using a CPD ELISA kit from Cell Biolabs (USA). The assay was based on a standardized solution of dsDNA (4 μg / mL) from each sample.

[0214] 2. UV-induced DNA damage in human reconstructed "full thickness" skin. In this model, highly standardized 3D reconstructed human skin tissue (EpiDerm FT) based on differentiated primary human keratinocytes and fibroblasts was provided by Mattek Europe (Slovakia). These tissues were grown on porous membranes and cultured in nutrient medium in 6-well plates in an incubator at 37°C and 5% CO2. Significant DNA damage was induced by placing the explants in a UV-solar simulator (SOL500 with UV filter H2) from Dr. Honle GmbH, Germany, which provided a combination of UVA and UVB radiation comparable to natural sunlight.

[0215] Four groups of four to five explants were treated with 100 mJ / cm 2 Once daily for two consecutive days, a control group of three explants without UV exposure was included to estimate the increase in DNA damage caused by UV treatment. Immediately after the last UV treatment, groups of explants were treated topically on the surface of the skin with 10 μL / explant of one of the following: 3.0% (w / w) of (R)-Mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate 1.0% (w / w) of (R)-Mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate 0.5% (w / w) of (R)-Mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate -Vehicle Treatment was repeated once daily for four consecutive days. On day -6, two 4 mm punch biopsies were taken and used as follows: One was used to estimate cell viability using the Resazurin Deep Blue Cell Viability Kit (Cat. No. 424702) from BioLegend (USA). Biopsies were incubated in 900 μL of fresh nutrient medium with 90 μL of Resazurin Deep Blue for 4 hours, after which the medium was collected and transferred to a black fluorescent 96-well plate (Thermo Scientific 137101), and fluorescence was measured on a Spectramax ID5 at 550 nm excitation and 590 nm emission to establish cell viability. The other biopsy was subjected to tissue homogenization in a GentleMACS™ tube from Miltenyi Biotec (Germany), followed by DNA extraction using the DNeasy® Blood & Tissue Kit (Qiagen, UK) according to standard protocols. - Double-stranded DNA (dsDNA) was quantified using the Invitrogen™ Quant-iT™ dsDNA Assay Kit (Cat. No. Q33130) from Thermo Fisher Scientific (Denmark). The amount of cyclobutane pyrimidine dimer (CPD) in each biopsy was quantified with a specific antibody using a CPD ELISA kit from Cell Biolabs (USA). The assay was based on a standardized solution of dsDNA (4 μg / mL) from each sample.

[0216] result: UV-induced DNA damage in human skin explants from middle-aged donors The levels of CPD for the three groups are shown in the table below. [Table 9]

[0217] Both concentrations of topical (R)-mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate showed highly biologically and statistically significant reductions in CPD indicating significant enhancement of DNA repair.

[0218] UV-induced DNA damage in human reconstructed "full-thickness" skin. The levels of CPD for the three groups are shown in the table below. [Table 10]

[0219] All concentrations of topical (R)-mandelic acid 4-hydroxy-3-methoxyphenyl-propanoate showed highly biologically and statistically significant reductions in CPD indicating significant enhancement of DNA repair.

[0220] Conclusion: The data show that topically administered (R)-mandelic acid 4-hydroxy-3-methoxyphenyl propanoate effectively penetrates the skin barrier and exerts a substantial enhancement of DNA repair. In addition to its relevance to skin aging, this effect indicates strong potential for the treatment of skin cancer and other conditions associated with DNA damage.

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Claims

1. A compound of general formula (I) or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 During the ceremony, * denotes the (S) or (R) enantiomer or any mixture thereof; z is an integer of 0, 1, 2, 3, 4, or 5; R １ are independently F, OH and C １ ~C ４ alkoxy; n is an integer of 0, 1, 2, 3, 4, or 5; R ２ are independently F, OH and C １ ~C ４ selected from alkoxy; R ３ is H and C １ ~C ８ alkyl.

2. 2. The compound of claim 1, wherein z is an integer from 0 to 4, preferably z is an integer from 0 to 3, and most preferably z is an integer from 1 to 3.

3. 3. The compound of claim 1 or 2, wherein n is an integer from 0 to 4, preferably n is an integer from 0 to 3, and most preferably n is an integer from 0 to 2.

4. 4. The compound according to claim 1, wherein n is an integer of 0 to 3 and z is an integer of 0 to 3.

5. R １ independently, OH and C １ ~C ３ 5. The compound of any one of claims 1 to 4, wherein the compound is selected from alkoxy.

6. R ２ independently, OH and C １ ~C ３ 5. The compound of any one of claims 1 to 4, wherein the compound is selected from alkoxy.

7. z is an integer from 0 to 3, and one or more R １ When the group is present, the group of formula (I) 【Chemistry 2】 7. The compound of claim 1, wherein the aryl group is located at the 3-, 4- and / or 5-position as shown in

8. n is an integer from 0 to 3, and one or more R ２ When the group is present, the group of formula (I) 【Transformation 3】 8. The compound of claim 1, wherein the aryl group is located at the 3-, 4- and / or 5-position as shown in

9. R ３ is H or C １ ~C ５ 9. The compound according to any one of claims 1 to 8, wherein said alkyl is selected as alkyl.

10. R ３ is H or C １ ~C ３ 9. The compound according to any one of claims 1 to 8, wherein said alkyl is selected as alkyl.

11. R ３ The compound of any one of claims 1 to 10, wherein is H.

12. 12. A compound according to any one of claims 1 to 11 for use as a pharmaceutical.

13. 13. A compound for use according to claim 12 for the treatment of skin diseases, rheumatic diseases, skin cancer and / or wounds.

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