Compound and application thereof in preparation of anti-aging drugs

Abstract

The present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, and its use in preventing, inhibiting, slowing, or alleviating aging.

Description

Technical Field

[0001] This application relates to the field of biomedicine, and more specifically to a class of compounds and their use in the preparation of anti-aging drugs. Background Technology

[0002] Aging, or senescence, refers to a gradual and irreversible biological process that occurs with increasing age, leading to the gradual decline of the body's structure and cellular function, resulting in physiological, biochemical, and morphological changes, weakened adaptability and resistance, and ultimately, death. To date, aging is the most important risk factor for various diseases, including cancer, coronary artery disease, Alzheimer's disease, Parkinson's disease, and chronic renal failure.

[0003] Aging includes immunosenescence, which can cause disorders in the lymphatic system and cellular function, increasing the risk of infections, tumors (including cancer), and other diseases. Anti-aging can inhibit immunosenescence, enhance immune function, and suppress tumor growth.

[0004] Telomeres, mitochondria, and inflammation are hallmarks of aging. With increasing age, imbalances in mitochondrial dynamics lead to mitochondrial dysfunction, accelerating aging progression and becoming closely associated with the occurrence and development of age-related diseases. The process of aging is inextricably linked to mitochondrial dysfunction.

[0005] Brown adipose tissue is part of adipose organs. The abundance of brown adipose tissue is associated with a lower prevalence of cardiometabolic diseases and a lower incidence of type II diabetes. However, the metabolic activity of brown adipose tissue gradually declines with age. This study assessed the aging status of brown adipose tissue in prematurely aging mice by detecting the transcriptional and protein expression levels of uncoupling proteinase 1 (UCP1), a key biomarker of brown adipose tissue activity.

[0006] Over the past few decades, the biomedical field has extensively explored and researched anti-aging mechanisms to prevent disease, and a variety of small molecule compounds and drugs with the potential to extend lifespan have been discovered. However, the field still needs to explore and discover more anti-aging drugs. Summary of the Invention

[0007] Provided are compounds of formula (I) or pharmaceutically acceptable salts thereof, or their use in the preparation of medicaments for the prevention, inhibition, slowing, or relief of aging, or for the prevention or treatment of aging-related diseases.

[0008]

[0009] in,

[0010] R 1 It can be unsubstituted or substituted with halogen, alkyl, or haloalkyl, aliphatic, aliphatic, aryl, aryl, heteroaryl, or heteroaryl;

[0011] R 2 It can be unsubstituted or substituted with halogen, alkyl, or haloalkyl, aliphatic, aliphatic, aryl, aryl, heteroaryl, or heteroaryl;

[0012] Q 1 For –L 1 Or –C(=A) 1 )–NH–L 1 ,in,

[0013] –L 1 The group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl is selected; and

[0014] =A 1 =O, =S, or =NH;

[0015] Q 5 Q 6 , and Q 7 Each is independently selected from hydrogen, halogen, alkoxy, heteroaryl, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 The group consisting of –L 0 The group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl is selected; and

[0016] A 4 It is carbon or nitrogen.

[0017] The compounds in some embodiments of this application can effectively improve metabolic homeostasis, slow down the decline in muscle strength caused by aging, and improve insulin sensitivity; and can enhance anti-tumor immunity and improve organ aging (including but not limited to improving kidney aging, increasing the number of lung mitochondria, increasing the browning tendency of brown adipose tissue, and enhancing the metabolic activity of brown adipose tissue). Attached Figure Description

[0018] Figure 1The results of (a) weekly body weight measurement, (b) grip strength measurement, (c) insulin resistance test (ITT), and (d) intraperitoneal glucose tolerance test (IPGTT) of mice in each group of Experiment 1 show that administration of the compound of this application can effectively improve metabolic instability in aged mice. (a) The horizontal axis is the duration after the first administration (weeks), and the vertical axis is body weight (g). ● represents the control group, and ▲ represents the #43 compound group; (b) The left side of the horizontal axis represents the control group, and the right side represents the #43 compound group. The vertical axis is grip strength (N); (c, d) The horizontal axis is the duration after injection (minutes), and the vertical axis is blood glucose (mmol / L). ● represents the control group, and ■ represents the #43 compound group.

[0019] Figure 2 In Experiment 2, after intraperitoneal inoculation of transplanted tumors and subsequent dissection, the following photographs were taken of mice in each group: (a) whole body, (b) spleen, (c) white adipose tissue, and (d) brown adipose tissue; and (e) comparison of the weights of the heart, liver, spleen, lungs, pancreas, thyroid gland, brain, cerebellum, brown adipose tissue, white adipose tissue, and beige adipose tissue. (a) The left image represents the control group, and the right image represents the #43 compound group. (b–d) Organs from the control group are placed on the left, and organs from the #43 compound group are placed on the right. (e) The horizontal axis, from left to right, represents the heart, liver, spleen, lungs, pancreas, thyroid gland, brain, cerebellum, brown adipose tissue, white adipose tissue, and beige adipose tissue; the vertical axis represents weight (g); gray bars represent the control group, and hollow bars represent the #43 compound group.

[0020] Figure 3 Example 2: After intraperitoneal inoculation of transplanted tumors and dissection, mice in each group underwent a comparison of (a) images and (b) images of β-galactosidase staining in their kidney cells. (a) Top: Control group; Bottom: Compound #43 group. (b) Horizontal axis: Left: Control group; Right: Compound #43 group; Vertical axis: Percentage of positive area (%).

[0021] Figure 4 In Experiment 2, after intraperitoneal inoculation of transplanted tumors and subsequent dissection, images of kidney tissue sections stained with Tom20 and DAPI were obtained: (a) and (b) a comparison of the proportion of Tom20-stained positive areas to the total area. (a) Top: Control group; Bottom: Compound #43 group; From left to right: TOM3 (red), DAPI (blue), and combined. (b) Horizontal axis: Left: Control group; Right: Compound #43 group; Vertical axis: Proportion of positive areas to the total area (%).

[0022] Figure 5In Experiment 2, after intraperitoneal inoculation of transplanted tumors and subsequent dissection, the kidney tissue sections stained with P21 and DAPI were compared in (a) and (b) with the proportion of P21-stained positive areas to the total area. (a) Top: Control group; Bottom: Compound #43 group; from left to right: P21 (red), DAPI (blue), and combined. (b) Horizontal axis: Left: Control group; Right: Compound #43 group; Vertical axis: Proportion of positive areas to the total area (%).

[0023] Figure 6 In Experiment 2, after intraperitoneal inoculation of transplanted tumors and subsequent dissection, lung tissue sections stained with Tom20 and DAPI were examined. (a) shows the images, and (b) compares the percentage of Tom20-stained positive areas in the total area. (a) Top: Control group; Bottom: Compound #43 group; From left to right: TOM3 (red), DAPI (blue), and combined. (b) Horizontal axis: Left: Control group; Right: Compound #43 group; Vertical axis: Percentage of positive areas (%).

[0024] Figure 7 In Experiment 2, after mice in each group were intraperitoneally inoculated with transplanted tumors and dissected, images of brown adipose tissue sections stained with hematoxylin and eosin (HE) (a) and immunohistochemically stained (b) and (c) were compared, showing a comparison of the proportion of positive areas showing UCP1 expression. (a, b) The top represents the control group, and the bottom represents the #43 compound group. (c) The horizontal axis represents the control group on the left and the #43 compound group on the right, and the vertical axis represents the proportion of positive areas (%).

[0025] Figure 8 Example 3: Photographs of mice in each group after 6 weeks of drug administration (a), (b), and (c) blood lipids. (a) Left: Control group; Right: Compound #43 group. (c) Horizontal axis from left to right: Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and total triglycerides (TG). (b, c) Solid column: Control group; Hollow column: Compound #43 group. (b, c) Vertical axis: Content (mmol / L).

[0026] Figure 9 Color photographs of the fundus of the retina of mice in each group of Experiment Example 4. The top, middle, and bottom views represent the normal group, control group, and compound 43 group, respectively. The left and right eye photographs are placed on the left and right sides, respectively. Detailed Implementation

[0027] To further illustrate the technical means and effects adopted by this application in order to achieve the intended purpose, the following detailed description of the specific implementation methods, structures, features and effects of this application is provided in conjunction with the accompanying drawings and preferred embodiments.

[0028] Terms and Definitions

[0029] As used herein, singular terms refer to one or more. For example, "element" or "a single element" both refer to one or more elements. As used herein, the term "multiple" refers to at least two.

[0030] As used herein, the term “approximately” refers to an approximation, within or near a range. When the term “approximately” is used in conjunction with a numerical range, it modifies the range by extending the boundaries above or below the provided numerical value. Generally, this document uses the term “approximately” to mean a numerical value that varies by 10% above or below the provided value. On the other hand, the term “approximately” means adding or subtracting 20% ​​of the numerical value of the number it modifies. For example, “approximately 50%” means within the range of 45%–55%. Numerical ranges referred to herein by endpoints include all integers and fractions contained within that range (e.g., “1 to 5” includes, but is not limited to, 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5). It should also be understood that all integers and fractions are considered to be modified by the term “approximately”.

[0031] As used herein, the terms “comprising,” “including,” or “containing,” as non-exclusive or open-ended terms, are intended to indicate that a combination (e.g., apparatus, composition, method, etc.) includes the listed elements (e.g., units of an apparatus, components of a composition, substantial steps of a method, etc.), but does not exclude other elements. As used herein, the term “substantially composed of” when used to define compositions and methods means excluding other elements that have any substantial effect on the combination for the stated purpose, but does not exclude other elements that do not substantially affect the basic and novel features of the invention. As used herein, the term “composed of” refers to a combination excluding other elements (units, components, substantial steps, etc.), but unless otherwise stated, it does not mean the exclusion of trace amounts of unavoidable impurities. Embodiments defined by each of these connecting terms are within the scope of the invention. As a particular embodiment, disclosure of technical solutions including the terms “comprising,” “including,” or “containing” should also be considered as simultaneous disclosure of corresponding technical solutions including the terms “substantially composed of” and “composed of.”

[0032] As used herein, the term "and / or" refers to and covers any and all possible combinations of one or more of the associated listed items. When used in a list of two or more items, the term "and / or" means that any one of the listed items may be included alone, or may include any combination of two or more listed items. For example, if a group, combination, or composition is described as including (or containing) components A, B, C, and / or D, then the composition may include A alone; include B alone; include C alone; include D alone; include a combination of A and B; include a combination of A and C; include a combination of A and D; include a combination of B and C; include a combination of B and D; include a combination of C and D; include a combination of A, B, and C; include a combination of A, B, and D; include a combination of A, C, and D; include a combination of B, C, and D; or include a combination of A, B, C, and D.

[0033] As used herein, the stereochemistry of chiral centers can be defined according to the conventions of those skilled in the art, i.e., using solid wedged bonds. "Indicate groups pointing outwards from the page (towards the reader) and use dashed wedge bonds." "Indicates a group that faces inwards (away from the reader). If such notation is used, it can be understood as indicating a specific, single stereoisomer of the group shown in the chemical structures herein. Any bond not specifically indicated by a solid or dashed wedge in this document should be considered as not specifically indicating whether the bond faces outwards, inwards, or on the page, but this does not preclude it from facing outwards or inwards where chemically permissible."

[0034] As used herein, the term "isomer" means a compound having the same molecular formula but differing in the bonding properties or sequence of its atoms or in the spatial arrangement of its atoms. Specifically, the term "stereoisomer" refers to isomers with different spatial arrangements of atoms; the term "enantiomer" refers to stereoisomers with one or more asymmetric centers, which are non-overlapping mirror images of each other; and the term "diastereomer" refers to stereoisomers that are not enantiomers but have opposite configurations at one or more asymmetric centers. When a compound has an asymmetric center, for example, if the carbon atom is bonded to four different groups, there can be a pair of enantiomers. Enantiomers can be characterized and designated as R-configuration or S-configuration by the absolute configuration of one or more asymmetric centers, or as dextrorotatory or levorotatory by the manner in which the molecule rotates in the plane of polarization. Chiral compounds can exist as individual enantiomers or as mixtures thereof, for example, as racemic mixtures. The compounds of this application may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. All stereoisomers of the compounds in this application, including but not limited to diastereomers, enantiomers, and transisomers, and mixtures thereof such as racemic mixtures, shall be considered as part of this application.

[0035] Implementation

[0036] Provided are compounds of formula (I) or pharmaceutically acceptable salts thereof. Provided are uses of compounds of formula (I) or pharmaceutically acceptable salts thereof in the prevention, inhibition, slowing, or relief of aging. Provided are uses of compounds of formula (I) or pharmaceutically acceptable salts thereof in the preparation of medicaments for the prevention, inhibition, slowing, or relief of aging. Provided are compounds of formula (I) or pharmaceutically acceptable salts thereof for the prevention, inhibition, slowing, or relief of aging. Provided are methods for the prevention, inhibition, slowing, or relief of aging, including the administration of compounds of formula (I) or pharmaceutically acceptable salts thereof.

[0037]

[0038] In some implementations, R 1 and R 2 Each is independently unsubstituted or substituted with halogen, alkyl, or haloalkyl, especially substituted with halogen or haloalkyl, even more especially substituted with fluorine or fluoroalkyl, especially substituted with fluorine or trifluoromethyl, aliphatic, aliphatic oxy, aryl, aryloxy, heteroaryl, or heteroaryloxy, especially aryl, aryloxy, heteroaryl, or heteroaryloxy, even more especially aryl or heteroaryl.

[0039] In some implementations, R 1 and R 2Each of the following is an aliphatic group, which is either unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, even more particularly substituted with a fluorine or fluoroalkyl group, especially substituted with a fluorine or trifluoromethyl group. In some embodiments, the aliphatic group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0040] In some implementations, R 1 and R 2 Each of the following is an aliphatic oxy group, individually unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, even more particularly substituted with a fluorine or fluoroalkyl group, and especially particularly substituted with a fluorine or trifluoromethyl group. In some embodiments, the aliphatic oxy group is selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0041] In some implementations, R 1 and R 2 Each is independently unsubstituted or substituted with halogen, alkyl, or haloalkyl, especially substituted with halogen or haloalkyl, even more particularly substituted with fluorine or fluoroalkyl, especially substituted with fluorine or trifluoromethyl, aryl or aryloxy, especially phenyl.

[0042] In some implementations, R 1 and R 2 Each aryl group is independently unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, even more particularly substituted with a fluorine or fluoroalkyl group, especially substituted with a fluorine or trifluoromethyl group. In some embodiments, the aryl group is selected from the group consisting of phenyl, naphthyl, anthraceneyl, and phenanthrene, particularly phenyl.

[0043] In some implementations, R 1 and R 2 Each of the following is an independent heteroaryl group, either unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, even more particularly substituted with a fluorine or fluoroalkyl group, and especially particularly substituted with a fluorine or trifluoromethyl group. In some embodiments, the heteroaryl group is pyrroleyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, etc. azole group, iso Azolyl, thiazolyl, pyridyl, pyranyl, thiaranyl, pyridazinyl, pyrimidinyl, pyrazinyl Azinyl, thiazinyl, indolyl, purinyl, quinolinyl, or isoquinolinyl, especially pyrroleyl, furanyl, or thiophenyl, and even more particularly thiophenyl.

[0044] In some implementations, R 1 and R2 Each aryl group is independently unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, even more particularly substituted with a fluorine or fluoroalkyl group, especially substituted with a fluorine or trifluoromethyl group, and is an aryl group. In some embodiments, the aryl group is selected from the group consisting of phenoxy, naphthyloxy, anthraceneyloxy, and phenanthreneoxy, particularly phenoxy.

[0045] In some implementations, R 1 and R 2 Each of these is an independent heteroaryloxy group, either unsubstituted or substituted with a halogen, alkyl, or haloalkyl group, particularly substituted with a halogen or haloalkyl group, and even more particularly substituted with a fluorine or fluoroalkyl group, especially substituted with a fluorine or trifluoromethyl group. In some embodiments, the heteroaryloxy group is pyrrolithoxy, furanoxy, thiophenoxy, pyrazoloxy, imidazoloxy, etc. Azoxy, iso- Azoxy, thiazoxy, pyridinoxy, pyranoxy, thiaranoxy, pyridazinoxy, pyrimidinoxy, pyrazinoxy Azineoxy, thiazineoxy, indoloxy, purineoxy, quinolineoxy, or isoquinolineoxy, especially pyrroleoxy, furanoxy, or thiophenoxy, and even more particularly thiophenoxy.

[0046] In some implementations, R 1 and R 2 Each is independently a phenyl substituted with a halogen or haloalkyl group, more particularly a phenyl substituted with a fluorine or fluoroalkyl group, and especially a phenyl substituted with a fluorine or trifluoromethyl group. In some embodiments, R 1 and R 2 Each is independently a halogen-substituted phenyl group. In some embodiments, R 1 and R 2 Each is independently a phenyl group substituted with a haloalkyl group. In some embodiments, the haloalkyl group is a trihalomethyl or a fluoroalkyl group, and particularly a trifluoromethyl group. In some embodiments, R 1 and R 2 Each is independently a fluorinated phenyl group. In some embodiments, R 1 and R 2 Each is an independent phenyl group substituted with a trifluoromethyl group.

[0047] In some embodiments, the halogen is fluorine, chlorine, bromine, or iodine, especially fluorine.

[0048] In some implementations, R 1 and R 2 Each is independent of the other: –Ph(–X) n (Right now In this context, each –X is independently a halogen or haloalkyl group, particularly a halogen or trihalomethyl group, particularly fluorine or trifluoromethyl; and n is 1, 2, 3, 4, or 5, particularly 1, 2, or 3. In some embodiments, –Ph(–X) is used. n The connection key is position 1, R 1 and R 2 Each element independently includes a –X located at the intermediate position (position 3 or 5), and optionally also includes at least one of a –X located at the adjacent position (position 2 or 6) and a –X located at the opposite position (position 4). In some embodiments, –Ph (–X) is used. n The connection key is position 1, R 1 and R 2 Each independently includes –X located at position 2; position 3; position 4; positions 2 and 3; positions 2 and 4; positions 2 and 5; positions 2 and 6; positions 3 and 4; positions 3 and 5; positions 2, 3 and 4; positions 2, 3 and 5; positions 2, 3 and 6; positions 2, 4 and 5; positions 2, 4 and 6; or positions 3, 4 and 5, wherein each –X is independently a halogen or haloalkyl, particularly a halogen or trihalomethyl, particularly fluorine or trifluoromethyl.

[0049] In some implementations, R 1 and R 2 Each is independently selected from the group consisting of fluorophenyl, difluorophenyl, trifluorophenyl, (trifluoromethyl)phenyl, di(trifluoromethyl)phenyl, and tri(trifluoromethyl)phenyl. Particularly in some embodiments, R 1 and R 2 Each of the following is independently selected from 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2,3-2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 3,4,5-trifluorophenyl, 2-(trifluoromethyl)phenyl, 3-(trifluoromethyl)phenyl, 4-(trifluoromethyl)phenyl, 2,3 The group consisting of -di(trifluoromethyl)phenyl, 2,4-di(trifluoromethyl)phenyl, 2,5-di(trifluoromethyl)phenyl, 2,6-di(trifluoromethyl)phenyl, 3,4-di(trifluoromethyl)phenyl, 3,5-di(trifluoromethyl)phenyl, 2,3-2,3,4-tri(trifluoromethyl)phenyl, 2,3,5-tri(trifluoromethyl)phenyl, 2,3,6-tri(trifluoromethyl)phenyl, 2,4,5-tri(trifluoromethyl)phenyl, 2,4,6-tri(trifluoromethyl)phenyl, and 3,4,5-tri(trifluoromethyl)phenyl.

[0050] In some implementations, R 1 It is 3-(trifluoromethyl)phenyl, R 2 It is 2,4,5-trifluorophenyl.

[0051] In some implementations, Q 5 Q 6 , and Q 7 Each is independently selected from hydrogen, halogen, alkoxy, heteroaryl, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 A group consisting of [a number of elements]. In some implementations, Q [is used]. 5 Q 6 , and Q 7 Each is independently selected from hydrogen, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 A group consisting of [a number of elements]. In some implementations, Q [is used]. 5 Q 6 , and Q 7 Each is independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, formyl, acetyl, propionyl, methoxycarbonyl, ethoxycarbonyl, and propoxycarbonyl. In some embodiments, Q 5 Q 6 , and Q 7 Each is independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, iodine, methoxy, and formyl groups. In some embodiments, Q 5 Q 6 , and Q 7 Each is independently selected from the group consisting of hydrogen, fluorine, chlorine, and methoxy. In some embodiments, the alkoxy group is methoxy, ethoxy, or propoxy. In some embodiments, –L 0 Choose from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl, and = A 1=O, =S, or =NH, especially =O or =S. In some embodiments, Q 1 For –L 0 In some implementations, Q 1 –C(=A1)–NH–L 0 .

[0052] In some implementations, –L 0 Choose free hydrogen, C 1-10 Alkyl, C 1-10 alkenyl, C 1-10 alkynyl, aryl C 1-10 Alkyl and heteroaryl C 1-10 The group consisting of alkyl groups, especially those composed of C 1-8 Alkyl, C 1-8 alkenyl, C 1-8 alkynyl, aryl C 1-8 Alkyl and heteroaryl C 1-8 The group consisting of alkyl groups, and more particularly those composed of C 1-6 Alkyl, C 1-6 alkenyl, C 1-6 alkynyl, aryl C 1-6 Alkyl and heteroaryl C 1-6 The group consisting of alkyl groups, and more particularly those composed of hydrogen and C 1-3 Alkyl, C 1-3 alkenyl, C 1-3 alkynyl, aryl C 1-3 Alkyl and heteroaryl C 1-3 The group consisting of alkyl groups. In some embodiments, –L 0 Choose C freely 4-10 Alkyl, C 4-10 alkenyl, C 4-10 alkynyl, aryl C 4-10 Alkyl and heteroaryl C 4-10 The group consisting of alkyl groups, especially those composed of C 4-8 Alkyl, C 4-8 alkenyl, C 4-8 alkynyl, aryl C 4-8 Alkyl and heteroaryl C 4-8 The group consisting of alkyl groups, and more particularly those composed of C 4-6 Alkyl, C 4-6 alkenyl, C 4-6 alkynyl, aryl C 4-6 Alkyl and heteroaryl C 4-6 The group consisting of alkyl groups. In some embodiments, –L 0 The group is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, allyl, propargyl, phenyl, and benzyl. In some embodiments, –L 0The alkyl group is selected from the group consisting of hydrogen, methyl, ethyl, allyl, propargyl, phenyl, and benzyl. In some embodiments, the alkyl group is methyl, ethyl, propyl, or isopropyl. In some embodiments, the alkenyl group is allyl. In some embodiments, the alkynyl group is propargyl.

[0053] In some implementations, Q 5 Q 6 , and Q 7 Each is independently selected from hydrogen, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 The group consisting of –L 0 Choose free hydrogen, C 1-10 Alkyl, C 1-10 alkenyl, C 1-10 alkynyl, aryl C 1-10 Alkyl and heteroaryl C 1-10 A group composed of alkyl groups.

[0054] In some implementations, Q 5 Q 6 , and Q 7 All are hydrogen.

[0055] In some implementations, A 4 It is carbon or nitrogen. In some embodiments, A 4 It is carbon. In some embodiments, A 4 It is nitrogen.

[0056] In some implementations, Q 1 For –L 1 Or –C(=A) 1 )–NH–L 1 In some implementations, –L 1 Choose from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl, and = A 1 =O, =S, or =NH, especially =O or =S. In some embodiments, Q 1 For –L 1 In some implementations, Q 1 For –C(=A) 1 )–NH–L 1 In some implementations, –L 1 Choose free hydrogen, C1-10 Alkyl, C 1-10 alkenyl, C 1-10 alkynyl, aryl C 1-10 Alkyl and heteroaryl C 1-10 The group consisting of alkyl groups, especially those composed of C 1-8 Alkyl, C 1-8 alkenyl, C 1-8 alkynyl, aryl C 1-8 Alkyl and heteroaryl C 1-8 The group consisting of alkyl groups, and more particularly those composed of C 1-6 Alkyl, C 1-6 alkenyl, C 1-6 alkynyl, aryl C 1-6 Alkyl and heteroaryl C 1-6 The group consisting of alkyl groups, and more particularly those composed of hydrogen and C 1-3 Alkyl, C 1-3 alkenyl, C 1-3 alkynyl, aryl C 1-3 Alkyl and heteroaryl C 1-3 The group consisting of alkyl groups. In some embodiments, –L 1 Choose C freely 4-10 Alkyl, C 4-10 alkenyl, C 4-10 alkynyl, aryl C 4-10 Alkyl and heteroaryl C 4-10 The group consisting of alkyl groups, especially those composed of C 4-8 Alkyl, C 4-8 alkenyl, C 4-8 alkynyl, aryl C 4-8 Alkyl and heteroaryl C 4-8 The group consisting of alkyl groups, and more particularly those composed of C 4-6 Alkyl, C 4-6 alkenyl, C 4-6 alkynyl, aryl C 4-6 Alkyl and heteroaryl C 4-6 The group consisting of alkyl groups. In some embodiments, –L 1 The group is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, allyl, propargyl, phenyl, and benzyl. In some embodiments, –L 1 The group consisting of hydrogen, methyl, ethyl, allyl, propargyl, phenyl, and benzyl is selected.

[0057] In some embodiments, the aryl group is phenyl, naphthyl, anthraceneyl, phenanthryl, or pyrene, particularly phenyl or naphthyl, and more particularly phenyl. In some embodiments, the heteroaryl group is pyrroleyl, furanyl, thiopheneyl, pyrazolyl, imidazoleyl, etc. azole group, iso Azolyl, thiazolyl, pyridyl, pyranyl, thiaranyl, pyridazinyl, pyrimidinyl, pyrazinyl Azinyl, thiazinyl, indoleyl, purineyl, quinolinyl, or isoquinolinyl.

[0058] In some embodiments, the aryl alkyl group is arylmethyl, arylethyl, or arylpropyl, particularly arylmethyl, and more particularly benzylmethyl. In some embodiments, the heteroaryl alkyl group is heteroarylmethyl, heteroarylethyl, or heteroarylpropyl. In some embodiments, the alkyl aryl group is methylaryl, ethylaryl, or propylaryl. In some embodiments, the alkyl heteroaryl group is methyl heteroaryl, ethyl heteroaryl, or propyl heteroaryl.

[0059] In some implementations, the C 1-10 C 1-8 C 1-6 or C 1-3 It can be C1. In some embodiments, the C... 1-10 C 1-8 C 1-6 or C 1-3 It can be C2. In some embodiments, the C... 1-10 C 1-8 C 1-6 or C 1-3 It can be C3. In some embodiments, the C... 1-10 C 1-8 C 1-6 C 4-10 C 4-8 or C 4-6 It can be C4. In some embodiments, the C... 1-10 C 1-8 C 1-6 C 4-10 C 4-8 or C 4-6 It can be C5. In some embodiments, the C... 1-10 C 1-8 C 1-6 C 4-10 C 4-8 or C 4-6 It can be C6. In some embodiments, the C... 1-10 C 1-8 C 4-10 Or C 4-8 It can be C7. In some embodiments, the C... 1-10 C 1-8 C 4-10 Or C 4-8 It can be C8. In some embodiments, the C... 1-10 Or C 4-10 It can be C9. In some embodiments, the C...1-10 Or C 4-10 It can be C 10 .

[0060] In some implementations, R 1 It is 3-(trifluoromethyl)phenyl; R 2 It is 2,4,5-trifluorophenyl; Q 1 Methyl; Q 5 Q 6 , and Q 7 Both are hydrogen; and A 4 It is carbon.

[0061] In some embodiments, formula (I) is one or more selected from the group consisting of formulas (I1), (I2), (I3), (I4), (I5), (I6), (I7), and (I8), or a racemic mixture thereof.

[0062]

[0063] In some embodiments, formula (I) is formula (I1) or formula (I8) or a racemic mixture thereof (composed of formula (I1) and formula (I8)). In some embodiments, formula (I) is formula (I2) or formula (I7) or a racemic mixture thereof (composed of formula (I2) and formula (I7)). In some embodiments, formula (I) is formula (I3) or formula (I6) or a racemic mixture thereof (composed of formula (I3) and formula (I6)). In some embodiments, formula (I) is formula (I4) or formula (I5) or a racemic mixture thereof (composed of formula (I4) and formula (I5)).

[0064] In some embodiments, the aging is organ aging, which may be selected from the group consisting of, for example, the eye, heart, liver, spleen, lungs, kidneys, pancreas, thyroid gland, thymus, brain, cerebellum, fat (e.g., brown fat, white fat, or beige fat), muscles, bones, joints, reproductive system (e.g., gonads, such as testes or ovaries), and neurons.

[0065] In some embodiments, the aging is selected from the group consisting of obesity, immune degeneration, reproductive degeneration, decreased organ mitochondrial expression, increased expression of organ aging markers, decreased brown adipose tissue activity, brown adipose tissue whitening, metabolic instability (e.g., glucose or lipid metabolism instability), decreased metabolic rate or activity, decreased muscle strength or mass, increased muscle fibrosis, decreased insulin sensitivity, increased risk of hyperglycemia, increased risk of hyperlipidemia, increased risk of hypertension, increased risk of heart disease, increased risk of kidney disease, increased risk of diabetes, increased risk of osteoporosis, macular degeneration, and increased risk of tumors (e.g., cancer).

[0066] In some embodiments, the prevention, inhibition, slowing down, or mitigation of aging is selected from the group consisting of reducing obesity, inhibiting or slowing down immune degeneration, inhibiting or slowing down reproductive degeneration, increasing organ mitochondrial expression, reducing organ aging marker expression, increasing brown adipose tissue activity, inhibiting or slowing down brown adipose tissue whitening, maintaining metabolic homeostasis (e.g., maintaining glucose homeostasis or maintaining lipid homeostasis), maintaining metabolic rate or metabolic activity, increasing muscle strength or mass, inhibiting or slowing down muscle fibrosis, improving insulin sensitivity, reducing the risk of hyperglycemia, reducing the risk of hyperlipidemia, reducing the risk of hypertension, reducing the risk of kidney disease, reducing the risk of diabetes, reducing the risk of osteoporosis, treating or preventing macular degeneration, and reducing the risk of tumors (e.g., cancer).

[0067] Example

[0068] Compound Synthesis

[0069] The reaction of compound (A) and compound (B) yields compound (I).

[0070]

[0071] In compounds of formula (A) and formula (B), R 1 R 2 Q 1 Q 5 Q 6 Q 7 A 4 The definition is the same as that in compound (I); X′ can be chlorine, bromine, or iodine.

[0072] The synthesis of the compound can be carried out in the presence of a base. The base can be sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, cesium carbonate, triethylamine, or triethylenediamine (i.e., 1,4-diazabicyclo[2.2.2]octane), especially triethylenediamine.

[0073] The synthesis of the compound can be carried out in the presence of a chiral auxiliary. Such chiral auxiliaries may be, for example, 1-(3,5-bis(trifluoromethyl)phenyl)-3-((1S)-(6-methoxyquinoline-4-yl)(5-vinylquinoline-2-yl)methyl)thiourea or 3-((3,5-bis(trifluoromethyl)phenyl)amino)-4-(((1S)-(6-methoxyquinoline-4-yl)(5-vinylquinoline-2-yl)methyl)amino)cyclobut-3-ene-1,2-dione.

[0074] The synthesis of compounds can be carried out in the presence of molecular sieves. The molecular sieves can be 3A, 4A, 5A, 10Z, or Y-type molecular sieves.

[0075] The synthesis of the compound can be carried out in the presence of a solvent. The solvent may be selected from aliphatic hydrocarbon solvents, aromatic hydrocarbon solvents, haloalkanes solvents, alcohol solvents, ester solvents, ketone solvents, sulfoxide solvents, amide solvents, nitrile solvents, and heterocyclic solvents, as well as combinations thereof, particularly from chloroform, dichloromethane, ethyl acetate, toluene, and acetonitrile, as well as combinations thereof.

[0076] The following illustrates specific preparation methods for some exemplary compounds of this application. Those skilled in the art should be able to deduce and implement the specific preparation methods for the compounds in various embodiments of this application by analogy with the specific preparation methods below and the prior disclosures mentioned above.

[0077] Preparation of compound 43# in Example 1

[0078] Compound 43# is compound (I), wherein R 1 It is 3-(trifluoromethyl)phenyl, R 2 It is 2,4,5-trifluorophenyl, Q 1 It is methyl, Q 5 Q 6 , and Q 7 Both are hydrogen, and A 4 The answer is C.

[0079]

[0080] 0.01 mmol of 1-(3,5-bis(trifluoromethyl)phenyl)-3-((1S)-(6-methoxyquinoline-4-yl)(5-vinylquininyl-2-yl)methyl)thiourea, 10 mg of 4A molecular sieve, triethylenediamine, 0.1 mmol of 3-chloro-1-methylindol-2-one (compound (A)) and 0.13 mmol of (E)-2-(3-(trifluoromethyl)benzoyl)-3-(2,4,5-trifluorophenyl)acrylonitrile (compound (B)) were dissolved in 3 mL of chloroform and the reaction was carried out with stirring at room temperature.

[0081] After the reaction was monitored to be complete by thin-layer chromatography (TLC), the reaction solution was concentrated by rotary evaporation. The crude product was purified by column chromatography (petroleum ether:ethyl acetate = 10:1 to 5:1) to give 4 g of product, which was compound 43#. Yield: 62%.

[0082] MS(ESI)m / z:C 26 H 15 F6N2O2, [M+H] + Calculated value: 501.1; Actual value: 501.14.

[0083] The obtained racemic compound 43# was separated to obtain four diastereomers of each other: compound 43-p1, compound 43-p2, compound 43-p3, and compound 43-p4, which are represented by one of the enantiomers.

[0084]

[0085] The characteristic data of compound 43-p1 are as follows:

[0086] 1 H NMR (600MHz, CDCl3) δ = 7.95 (s, 1H), 7.90-7.83 (m, 2H), 7.61-7.55 (m, 1H), 7.54-7.47 (m, 1H), 7.26- 7.20(m,1H),7.13-7.06(m,2H),7.05-6.97(m,1H),6.63(d,J=7.6Hz,1H),4.05(s,1H),3.17(s,3H).

[0087] 13 C NMR (150MHz, CDCl3) δ = 185.7, 167.3, 156.8 (ddd, 1 J C–F =246.8,9.6,2.8Hz),150.4(ddd, 1 J C–F =253.3,14.2,12.3Hz),146.8(ddd, 1 J C–F =246.2,12.6,3.6Hz),144.2,134.4,132.0,131.8(q, 2 J C–F =33.3Hz), 131.0(q, 3 J C–F =3.5Hz),130.1,129.6,126.0(q, 3 J C–F =4.0Hz), 123.1(d, 1 J C–F =272.8Hz),122.85,121.35,120.86,118.94(ddd, 2 J C–F =20.8,4.3,1.5Hz),113.2(ddd, 2 J C–F =27.0,10.2,1.8Hz),112.9,109.9,105.9(dd, 2 J C–F=27.3,21.2Hz),41.9(d, 3 J C–F =1.7Hz), 38.8, 30.8, 27.0.

[0088] 19 F NMR (564MHz, CDCl3) δ = -113.403, -130.439, -140.363, -140.464.

[0089] The characteristic data of compound 43-p2 are as follows:

[0090] 1 H NMR (600MHz, CDCl3) δ = 7.95 (s, 1H), 7.89-7.82 (m, 2H), 7.60-7.55 (m, 1H), 7.54-7.48 (m, 1H), 7.26-7.2 0(m,1H),7.13-7.06(m,2H),7.05-6.97(m,1H),6.62(d,J=7.6Hz,1H),4.07-4.03(m,1H),3.16(s,3H).

[0091] 19 F NMR (564MHz, CDCl3) δ = -113.348, -130.393, -140.381.

[0092] The characteristic data of compound 43-p3 are as follows:

[0093] 1 H NMR (600MHz, CDCl3) δ = 7.93 (s, 1H), 7.76 (br t,J=9.2Hz,2H),7.47(t,J=8.0Hz,1H),7.27(s,2H),6.98-6.90(m,2H),6.86(t,J=7.6Hz,1H),6.68(d,J=7.6Hz,1H),4.16(s,1H),3.38(s,3H).

[0094] 19 F NMR (564MHz, CDCl3) δ = -115.516, -131.822, -140.953, -141.008.

[0095] The characteristic data of compound 43-p4 are as follows:

[0096] 1H NMR (600MHz, CDCl3) δ = 7.97 (s, 1H), 7.81 (t, J = 8.7Hz, 2H), 7.51 (t, J = 7.9Hz, 1H), 7.42-7.28 (m, 2H ),7.02-6.94(m,2H),6.90(dt,J=0.7,7.7Hz,1H),6.72(d,J=7.4Hz,1H),4.19(s,1H),3.42(s,3H).

[0097] 19 F NMR (564MHz, CDCl3) δ = -115.516, -131.804, -140.990.

[0098] Anti-aging activity of compounds

[0099] Example 1: Compound 43 improves metabolic homeostasis in aged rats

[0100] Grouping and administration of experimental animals

[0101] Eighteen-month-old commercial mice were prepared and divided into a control group and a compound 43 group according to their body weight.

[0102] - Control group: 100 μL of a solvent consisting of 5% DMSO, 15% Tween 80 and 80% PBS was injected into mice via the tail vein at the time of administration.

[0103] -43# compound group: Based on the mouse body weight and the mouse drug dosage (15 mg / kg mouse body weight), the corresponding weight of 43# compound was dissolved in 100 μL of solvent to obtain the 43# compound drug solution; it was injected into the mice via the tail vein during administration.

[0104] Administer once a week for 22 weeks.

[0105] 1.1 Weight Measurement

[0106] Mice were weighed and their weight recorded weekly. Unpaired t-tests were used to compare the two groups, with p < 0.05 considered statistically significant.

[0107] The results are as follows Figure 1 As shown in (a), the body weight of the 43# compound group was lower than that of the control group.

[0108] 1.2 Grip strength measurement

[0109] Grip strength was measured in two groups of mice after the drug administration period using a small animal grip strength meter, and the measurements were recorded. Unpaired t-tests were used to compare the two groups, with p < 0.05 considered statistically significant.

[0110] The results are as follows Figure 1 As shown in (b), mice in the compound 43 group had higher grip strength than the control group. Administration of compound (I) to subjects improved their muscle strength and slowed age-related muscle weakness decline.

[0111] 1.3 Insulin Tolerance Test

[0112] After the dosing cycle, mice were fasted for 4 hours but allowed free access to water. They were then injected intraperitoneally with 1 U / kg of insulin. Blood samples were collected from the tail vein at 0, 15, 30, 60, and 120 minutes post-injection to measure and record blood glucose levels. Two-way Anova was used for comparison between the two groups, with p < 0.05 considered statistically significant.

[0113] The results are as follows Figure 1 As shown in (c), the blood glucose levels in mice in the compound 43 group were lower than those in the control group over 120 minutes. Administration of compound (I) to subjects improved their insulin sensitivity.

[0114] 1.4 Glucose tolerance test

[0115] After the drug administration cycle, mice were fasted for 14 hours but allowed free access to water. They were then injected intraperitoneally with 1 g / kg of 20% glucose solution. Blood samples were collected from the tail vein at 0, 15, 30, 60, and 120 minutes post-injection to measure and record blood glucose levels. Two-way ANOVA was used for comparison between the two groups, with p < 0.05 considered statistically significant.

[0116] The results are as follows Figure 1 As shown in (d), the blood glucose levels in mice in the compound 43 group were lower than those in the control group within 120 minutes. Administration of compound (I) to subjects improved their ability to regulate blood glucose concentration.

[0117] Experimental Example 2: Compound 43# enhances anti-tumor immunity and improves parenchymal organ aging in aged mice.

[0118] Grouping and administration of experimental animals

[0119] Same as Experiment 1.

[0120] 2.1 Intraperitoneal inoculation of transplanted tumors

[0121] After the dosing cycle was completed, aged mice in both the control and experimental groups were intraperitoneally injected with MC38 mouse colon cancer cells, with a cell count of 8 × 10⁻⁶. 5 indivual.

[0122] The mice were anesthetized and euthanized 14 days after inoculation.

[0123] The size of the abdominal tumor and spleen was recorded by photograph, and the weight of the solid organs of the two groups of mice was recorded.

[0124] The results are as follows Figure 2 As shown in (a) to (e). Figure 2 (a) shows that the tumors in mice in the 43# compound group were significantly reduced compared to the control group; Figure 2 (b) shows that the number of splenic tumors in mice in the 43# compound group was reduced compared with that in control mice; Figure 2 (c) and (e) show that the weight of white adipose tissue in the abdominal cavity of mice in the compound 43 group was significantly lower than that in the control group; Figure 2 (d) shows that the brown adipose tissue in mice in the compound 43 group showed a greater tendency to brown compared to the control group. Administration of compound (I) to elderly subjects enhanced their antitumor immunity.

[0125] Kidney samples were taken from the euthanized mice in the following experimental examples 2.2 and 2.3; lung samples were taken in the following experimental example 2.4; and brown adipose tissue samples were taken in the following experimental examples 2.5 and 2.6.

[0126] 2.2 β-galactosidase staining of kidney cells

[0127] The aforementioned kidney samples were serially frozen sectioned to a thickness of 10 μm. The frozen sections were thawed at room temperature for 10 minutes, and the tissue to be tested was circled with an immunohistochemical pen. An appropriate amount of β-galactosidase staining fixative was added to the tissue, and the sections were fixed at room temperature for 20 minutes. The sections were then washed three times with PBS for 5 minutes each time, and then placed in a humidified chamber. An appropriate amount of β-galactosidase staining working solution was added, and the sections were incubated overnight at 37°C in a CO2-free incubator. After tissue staining, the staining solution was removed, and the sections were washed twice with PBS and twice with pure water. Nucleotide red staining solution was added for 3 minutes, followed by three washes with water. The sections were dehydrated twice with anhydrous ethanol for 5 minutes each time, cleared with xylene for 5 minutes, and then mounted with neutral resin.

[0128] Image J was used to perform statistical analysis on the positive regions, and an unpaired t-test was used, with p < 0.05 considered statistically significant. The results are as follows: Figure 3 As shown in (a) and (b), the number of β-galactosidase-positive areas in the kidneys of mice in the compound 43 group was significantly reduced compared to the control group. Administration of compound (I) to subjects improved renal aging.

[0129] 2.3 Immunofluorescence staining of kidney tissue sections

[0130] The aforementioned kidney samples were serially frozen sectioned to a thickness of 10 μm. The frozen sections were thawed at room temperature for 10 minutes, fixed in ice-cold methanol at -20°C for 10 minutes, and washed three times with PBS. 3% hydrogen peroxide solution was added, and the sections were washed three times with PBS after 10 minutes. Immunostaining blocking solution was added for blocking at room temperature, and the blocking solution was discarded after 1 hour. Primary antibody (Tom20 or P21) was added and incubated overnight at 4°C. The primary antibody was then recovered, and the sections were washed three times with PBS. Goat anti-rabbit fluorescent secondary antibody was added and incubated at room temperature for 1 hour, followed by three washes with PBS. DAPI staining was added at room temperature. After 5 minutes, the sections were mounted using anti-fluorescence quenching mounting solution.

[0131] Images were taken and recorded under a fluorescence microscope. Image J was used to perform statistical analysis on the positive areas. An unpaired t-test was used, and p < 0.05 was considered statistically significant.

[0132] Tom20 staining results of kidney tissue sections Figure 4 As shown in (a) and (b), the number of kidney mitochondria in mice in the compound 43 group was significantly increased compared to the control group. Administration of compound (I) to subjects increased the number of kidney mitochondria.

[0133] P21 staining results of kidney tissue sections Figure 5 As shown in (a) and (b), the expression of the aging marker P21 was significantly lower in the compound 43 group compared to the control group. Administration of compound (I) to subjects improved renal aging.

[0134] 2.4 Immunofluorescence staining of lung tissue sections

[0135] The aforementioned lung samples were serially frozen sectioned to a thickness of 10 μm. The frozen sections were thawed at room temperature for 10 minutes, fixed in ice-cold methanol at -20°C for 10 minutes, and washed three times with PBS. 3% hydrogen peroxide solution was added, and the sections were washed three times with PBS after 10 minutes. Immunostaining blocking solution was added and the sections were blocked at room temperature for 1 hour, then the blocking solution was discarded. Primary antibody (Tom20) was added and incubated overnight at 4°C. The primary antibody was then recovered, and the sections were washed three times with PBS. Goat anti-rabbit fluorescent secondary antibody was added and incubated at room temperature for 1 hour, followed by three washes with PBS. DAPI staining was added at room temperature. After 5 minutes, the sections were mounted using anti-fluorescence quenching mounting solution.

[0136] Images were taken and recorded under a fluorescence microscope. Image J was used to perform statistical analysis on the positive areas. An unpaired t-test was used, and p < 0.05 was considered statistically significant.

[0137] Tom20 staining results of lung tissue sections are as follows Figure 6 As shown in (a) and (b), the number of lung mitochondria in mice in the compound 43 group was significantly increased compared to the control group. Administration of compound (I) to subjects increased the number of lung mitochondria.

[0138] 2.5 Brown fatty hematoxylin-eosin (HE) staining

[0139] The aforementioned brown fat samples were fixed in 4% paraformaldehyde solution and then transferred to 10% neutral formalin solution. The brown fat samples were then sequentially immersed in 70% ethanol once, 85% ethanol once, 95% ethanol twice, anhydrous ethanol twice, xylene three times, and paraffin three times for dehydration, followed by embedding and sectioning.

[0140] The sections were placed in xylene twice to dewax, and then in anhydrous ethanol twice, 95% ethanol once, 85% ethanol once, and 75% ethanol once to rehydrate.

[0141] After rehydration, the sections were stained with hematoxylin for 14 seconds, then with another hematoxylin solution for 1 minute, and washed with water for 5 minutes. Differentiation was then performed with differentiation solution for 3-5 seconds, followed by washing with water for 1 minute. The sections were treated with blueing solution for 7 seconds, followed by washing with water for 20 seconds. The sections were then immersed in 95% ethanol for 1 minute, followed by staining with eosin for 14 seconds. The stained sections were then immersed in anhydrous ethanol twice for 2 minutes each, followed by immersion in xylene twice for 2 minutes each. Finally, the sections were mounted with neutral resin.

[0142] The results are as follows Figure 7 As shown in (a). With Figure 2 (d) also shows that the brown adipose tissue in mice in the compound 43 group was more prone to browning than that in the control group. Administration of compound (I) to subjects increased their brown adipose tissue's tendency to brown.

[0143] 2.6 Immunohistochemical staining of brown fat paraffin sections

[0144] The preparation and rehydration of brown fatty paraffin sections were the same as in Experiment 2.5.

[0145] After rehydration, the slides were microwaved on high for 15 minutes for antigen retrieval, and then washed three times with PBS for 5 minutes each time.

[0146] The tissue area to be tested was circled with an immunohistochemical pen, and immunostaining blocking solution was added. The area was blocked at room temperature for 1 hour, then the blocking solution was discarded. Primary antibody was added and incubated overnight at 4°C. The primary antibody was then recovered, and the tissue was washed three times with PBS for 5 minutes each time. Goat anti-rabbit HRP-conjugated secondary antibody was then added and incubated at room temperature for 1 hour, followed by three washes with PBS for 5 minutes each time.

[0147] Observe under a microscope and add DAB colorimetric solution. Once a positive result appears, immediately place the container in water to terminate the reaction.

[0148] After terminating the reaction, the sections were stained with hematoxylin for 1 minute and washed with water for 5 minutes. The stained sections were then immersed in anhydrous ethanol twice for 2 minutes each, followed by immersion in xylene twice for 2 minutes each. The sections were then mounted with neutral resin.

[0149] Images were taken and recorded under a fluorescence microscope. Image J was used to perform statistical analysis on the positive areas. An unpaired t-test was used, and p < 0.05 was considered statistically significant.

[0150] The results are as follows Figure 7 As shown in (b) and (c), the expression of UCP1 in brown adipose tissue of mice in compound 43 group was significantly increased compared with that in control mice. Administration of compound (I) to subjects improved the aging of their brown adipose tissue.

[0151] Example 3: Compound 43# improves body weight and metabolic status in type II diabetic mice

[0152] Grouping and administration of experimental animals

[0153] Prepare db / db mice and divide them equally into a control group and a 43# compound group according to their body weight.

[0154] - Control group: 200 μL of a solvent consisting of 5% DMSO, 15% Tween 80 and 80% PBS was injected into mice via the tail vein at the time of administration.

[0155] -43# compound group: Based on the mouse body weight and the mouse drug dosage (15 mg / kg mouse body weight), the corresponding weight of 43# compound was dissolved in 200 μL of solvent to obtain 43# compound drug solution; it was injected into the mice via the tail vein during administration.

[0156] Mice were fed a normal, conventional fat diet and administered the drug once a week for 6 weeks.

[0157] 3.2 Mouse body shape

[0158] Figure 8 (a) shows photographs of db / db mice in the control group and the 43# compound group 6 weeks after drug administration. The left side represents the control group, and the right side represents the 43# compound group. Figure 8 As shown in (a), mice in the compound 43 group were significantly fatter than mice in the control group. Administration of compound (I) to subjects significantly reduced their body weight.

[0159] 3.3 Blood glucose and blood lipid measurements

[0160] Blood was collected by cutting the tip of the mouse tail 2mm with scissors, and blood glucose (Yuwell Blood Glucose Meter 580) and blood lipids (Ultra-Tech Blood Lipid Analyzer PFS-30) were analyzed.

[0161] The results are as follows Figure 8As shown in (b) and (c). Mice in the compound 43 group had significantly lower levels of blood glucose, total cholesterol, high-density lipoprotein cholesterol, and total triglycerides compared to the control group. Administration of compound (I) to subjects significantly reduced their blood glucose, total cholesterol, high-density lipoprotein cholesterol, and total triglycerides.

[0162] Example 4: Compound #43 improves fundus lesions in mice

[0163] Grouping and administration of experimental animals

[0164] Prepare normal mice as the normal group, and T... reg Cellular Abdh5 knockout mice were randomly assigned to either the control group or the 43# compound group.

[0165] T was prepared using Cre-LoxP technology. reg Abdh5 knockout mice were created using Foxp3 cells. Cre Mice (gifted by Professor Ye Lilin of Army Medical University) and Abdh5 flox / flox Mice (Cyagen (Suzhou) Biotechnology) were mated and positive mice were selected from the F1 generation to obtain T mice. reg Abdh5 knockout mice.

[0166] - Normal group: Normal mice were administered 100 μL of a solvent consisting of 5% DMSO, 15% Tween 80 and 80% PBS via tail vein injection.

[0167] -Control group: using T reg Abdh5 knockout mice were administered the solvent via tail vein injection of 100 μL.

[0168] Compound group -43#: using T reg In Abdh5 knockout mice, the corresponding weight of compound 43 was dissolved in 100 μL of solvent according to the mouse body weight and the drug dosage (15 mg / kg mouse body weight) to obtain the compound 43 drug solution; the solution was then injected into the mice via the tail vein.

[0169] Administer once a week for 4 months.

[0170] 4.1 Acquisition of color photographs of the mouse retina and fundus

[0171] Mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital solution at a dose of 50 mg / kg. The pupils of both eyes were dilated using compound tropicamide eye drops (0.5% tropicamide and 0.5% phenylephrine hydrochloride). Color photographs of the mouse retina were acquired using a bright-field imaging system with the optic nerve as the center, using the Phoenix MicronIV small animal retinal imaging system (Phoenix, USA). 1.7% sodium hyaluronate gel was used during imaging to prevent corneal dryness.

[0172] The results are as follows Figure 9 As shown. No obvious lesions were observed in the fundus of the normal group mice, while numerous yellowish-white lesions were visible in the fundus of the control group mice, indicating significant fundus lesions. In contrast, the fundus lesions in the compound 43# group mice were significantly alleviated. Administration of compound (I) to the subjects significantly improved their fundus lesions.

[0173] The above embodiments are merely preferred embodiments of this application and should not be construed as limiting the scope of protection of this application. Any non-substantial changes and substitutions made by those skilled in the art based on this application shall fall within the scope of protection claimed by this application.

Claims

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention, inhibition, slowing down, or relief of aging. in, R 1 It is an aliphatic group, aliphatic oxy group, aryl group, aryl oxy group, heteroaryl group, or heteroaryl oxy group that is unsubstituted or substituted with halogen, alkyl, or haloalkyl. R 2 It is an aliphatic group, aliphatic oxy group, aryl group, aryl oxy group, heteroaryl group, or heteroaryl oxy group that is unsubstituted or substituted with halogen, alkyl, or haloalkyl. Q 1 For –L 1 Or –C(=A) 1 )–NH–L 1 ,in, –L 1 The group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl is selected; and =A 1 =O, =S, or =NH; Q 5 Q 6 , and Q 7 Each is independently selected from hydrogen, halogen, alkoxy, heteroaryl, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 The group consisting of –L 0 The group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylalkyl, and heteroarylalkyl is selected; and A 4 It is carbon or nitrogen.

2. The use as described in claim 1, characterized in that, The aging referred to is organ aging, and the organs are selected from the group consisting of the eyes, heart, liver, spleen, lungs, kidneys, pancreas, thyroid gland, thymus, brain, cerebellum, brown fat, white fat, beige fat, muscles, bones, joints, reproductive system, and neurons.

3. The use as described in claim 1 or 2, characterized in that, The aging criteria were selected from the group consisting of obesity, immune degeneration, reproductive degeneration, decreased mitochondrial expression in organs, increased expression of organ aging markers, decreased brown adipose tissue activity, brown adipose tissue whitening, metabolic instability, decreased metabolic rate or activity, decreased muscle strength or mass, increased muscle fibrosis, decreased insulin sensitivity, increased risk of hyperglycemia, increased risk of hyperlipidemia, increased risk of hypertension, increased risk of heart disease, increased risk of kidney disease, increased risk of diabetes, increased risk of osteoporosis, macular degeneration, and increased risk of tumors.

4. The use as described in any one of claims 1 to 3, characterized in that, R 1 It is substituted with halogen or haloalkyl, and more particularly with fluorine or fluoroalkyl, especially with fluorine or trifluoromethyl, aliphatic, aliphatic oxy, aryl, aryloxy, heteroaryl, or heteroaryloxy. Optionally, R 1 It is substituted with halogen or haloalkyl, and more particularly with fluorine or fluoroalkyl, especially with fluorine or trifluoromethyl, aryl or heteroaryl, particularly phenyl.

5. The use as described in any one of claims 1 to 4, characterized in that, R 1 For –Ph(–X) n , in, Each –X is independently a halogen or haloalkyl group, particularly a halogen or trihalomethyl group, especially a fluorine or trifluoromethyl group; and n is 1, 2, 3, 4, or 5, especially 1, 2, or 3; Optionally, use –Ph(–X) n The connection key is position 1, R 1 It includes –X located between positions, and optionally also includes at least one of –X located adjacent to and –X located opposite to positions.

6. The use as described in any one of claims 1 to 5, characterized in that, R 2 It is substituted with halogen or haloalkyl, and more particularly with fluorine or fluoroalkyl, especially with fluorine or trifluoromethyl, aliphatic, aliphatic oxy, aryl, aryloxy, heteroaryl, or heteroaryloxy. Optionally, R 2 It is substituted with halogen or haloalkyl, and more particularly with fluorine or fluoroalkyl, especially with fluorine or trifluoromethyl, aryl or heteroaryl, particularly phenyl.

7. The use as described in any one of claims 1 to 6, characterized in that, R 2 For –Ph(–X) n , in, Each –X is independently a halogen or haloalkyl group, particularly a halogen or trihalomethyl group, especially a fluorine or trifluoromethyl group; and n is 1, 2, 3, 4, or 5, especially 1, 2, or 3; Optionally, use –Ph(–X) n The connection key is position 1, R 2 It includes –X located between positions, and optionally also includes at least one of –X located adjacent to and –X located opposite to positions.

8. The use as described in any one of claims 1 to 7, characterized in that, Q 5 Q 6 , and Q 7 Each is independently selected from hydrogen, –C(=O)–L 0 –O–C(=O)–L 0 –NH–C(=O)–L 0 –S(=O)2–L 0 –NH–S(=O)2–L 0 –NH–C(=O)–NH–L 0 , and –NH–C(=S)–NH–L 0 The group consisting of –L 0 Choose free hydrogen, C 1-10 Alkyl, C 1-10 alkenyl, C 1-10 alkynyl, aryl C 1-10 Alkyl and heteroaryl C 1-10 The group consisting of alkyl groups; and / or A 4 The answer is C; Optionally, –L 0 Choose free hydrogen, C 1-3 Alkyl, C 1-3 alkenyl, C 1-3 alkynyl, aryl C 1-3 Alkyl and heteroaryl C 1-3 The group consisting of alkyl groups; Optionally, Q 5 Q 6 , and Q 7 All are hydrogen.

9. The use as described in any one of claims 1 to 8, characterized in that, –L 1 Choose free hydrogen, C 1-10 Alkyl, C 1-10 alkenyl, C 1-10 Alkyne group, azide C 1-10 Alkyl, aryl C 1-10 Alkyl and heteroaryl C 1-10 The group consisting of alkyl groups; Optionally, –L 1 Choose C freely 4-10 Alkyl, C 4-10 alkenyl, C 4-10 Alkyne group, azide C 4-10 Alkyl, aryl C 4-10 Alkyl and heteroaryl C 4-10 The group consisting of alkyl groups; Optionally, Q 1 For –L 1 , and –L 1 It is a methyl group.

10. The use as described in any one of claims 1 to 9, characterized in that, R 1 It is 3-(trifluoromethyl)phenyl; R 2 It is 2,4,5-trifluorophenyl; Q 1 It is methyl; Q5, Q6, and Q7 are all hydrogen; and A4 is carbon.

Images