Application of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving indications of aging

Abstract

This invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving age-related indications. The OICR-9429 provided by this invention exhibits multiple beneficial technical effects in improving age-related indications, effectively alleviating the inflammatory response caused by pyroptosis during aging, and improving the immune function and anti-infection ability of aging organisms. OICR-9429 also has a significant therapeutic effect on sepsis, inhibiting the rise in inflammation levels caused by infection, improving the survival rate and prognosis of elderly patients. Long-term intervention with OICR-9429 can continuously reduce the chronic inflammatory state of aging organisms, increase muscle mass, enhance motor ability, prolong life, and reduce the occurrence of various age-related diseases. This invention lays the foundation for the development of products that comprehensively intervene in aging and related functional decline, and has important application value in improving the quality of life of the elderly.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more particularly to the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving indications of aging. Background Technology

[0002] Chronic low-grade inflammation has been identified as a core mechanism driving the occurrence and progression of many age-related diseases. This persistent, low-level systemic inflammatory state is closely related to the progressive dysfunction of the immune system, together forming the basic pathological environment of functional decline and increased disease risk in aging organisms.

[0003] Current clinical strategies for intervening in age-related inflammation primarily rely on traditional medications such as nonsteroidal anti-inflammatory drugs (NSAIDs), immunomodulators, and glucocorticoids. These drugs work by nonspecifically downregulating inflammatory cytokine networks or broadly suppressing immune cell activity. While they can alleviate some inflammatory symptoms in the short term, their limitations in long-term use are becoming increasingly apparent. NSAIDs are prone to causing gastrointestinal mucosal damage and renal dysfunction; the broad immunosuppressive effects of glucocorticoids significantly increase the risk of infection and metabolic disorders; and while biologics such as anti-TNF-α antibodies are effective for specific inflammatory diseases, they are expensive and may pose potential risks due to excessive suppression of immune surveillance. More fundamentally, these methods are not designed to target the inherent inflammatory mechanisms of aging itself; their effects often remain at the symptom level and cannot reverse or halt the intrinsic changes in the age-related immune system.

[0004] The shortcomings of existing treatments are mainly reflected in several dimensions. First, the effects are short-lived and unsustainable, making it difficult to achieve long-term, stable regulation of the chronic process of aging, and the efficacy diminishes over time. Second, the selectivity of targets is generally poor; while intervening in inflammation, drugs frequently interfere with normal physiological immune defense, tissue repair, and other key processes, resulting in a narrow therapeutic window and side effects becoming a major obstacle to long-term application.

[0005] Recent studies have revealed that pyroptosis plays a crucial role in the occurrence and persistence of age-related chronic low-grade inflammation. This process can be non-infectiously triggered, leading to altered cell membrane permeability and the release of large amounts of pro-inflammatory mediators, which continuously exacerbate local and systemic inflammation. However, existing anti-inflammatory drugs primarily target classical inflammatory pathways or immune cells, lacking effective and specific regulatory capabilities against pyroptosis, a specific mechanism of inflammatory amplification. Therefore, how to specifically intervene in age-related pyroptosis pathways without significantly suppressing overall immune function has become a critical issue that urgently needs to be addressed in the field of anti-aging. Summary of the Invention

[0006] To address the aforementioned problems in the prior art, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving indications of aging.

[0007] In a first aspect, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for anti-aging, wherein the structural formula of OICR-9429 is as follows:

[0008] .

[0009] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for anti-aging, preferably, the OICR-9429 or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0010] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of anti-aging products is preferably described in that the effective concentration of OICR-9429 or a pharmaceutically acceptable salt thereof is 3 mg / kg to 30 mg / kg.

[0011] More preferably, the effective concentration of the OICR-9429 or a pharmaceutically acceptable salt thereof is 5 mg / kg.

[0012] In this invention, the effective concentration of OICR-9429 is the applicable dose for mice. Those skilled in the art can obtain the equivalent dose applicable to other species, including but not limited to humans, rats, rabbits, dogs, cats, and pigs, by conventional technical means such as the body surface area conversion method.

[0013] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for anti-aging, preferably, the products comprising pharmaceuticals.

[0014] More preferably, the drug further includes pharmaceutically acceptable excipients.

[0015] More preferably, the excipients include sulfobutyl ether-β-cyclodextrin.

[0016] More preferably, the dosage form of the drug is a powder, tablet, granule, capsule, solution, emulsion or suspension.

[0017] Secondly, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for prolonging lifespan and / or delaying aging, wherein the structural formula of OICR-9429 is [insert structural formula here].

[0018] .

[0019] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for prolonging life and / or delaying aging is preferably used to achieve any one or more of the following objectives: (1) improving age-related decline in motor function; (2) reducing age-related inflammatory response; (3) reducing age-related lesions; and (4) improving age-related decline in disease resistance.

[0020] More preferably, the improvement of age-related decline in motor function includes any one or more of the following: (1) increasing muscle mass; (2) enhancing muscle strength; (3) improving motor coordination; (4) improving motor balance; and (5) improving motor endurance.

[0021] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for prolonging life and / or delaying aging is preferably described in that OICR-9429 or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0022] According to the present invention, the effective concentration of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for prolonging life and / or delaying aging is preferably 3 mg / kg to 30 mg / kg.

[0023] More preferably, the effective concentration of the OICR-9429 or a pharmaceutically acceptable salt thereof is 5 mg / kg.

[0024] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for prolonging life and / or delaying aging, preferably, the products comprise pharmaceuticals.

[0025] More preferably, the drug further includes pharmaceutically acceptable excipients.

[0026] More preferably, the excipients include sulfobutyl ether-β-cyclodextrin.

[0027] More preferably, the dosage form of the drug is a powder, tablet, granule, capsule, solution, emulsion or suspension.

[0028] Thirdly, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving the motor function of older individuals, wherein the structural formula of OICR-9429 is [insert structural formula here].

[0029] .

[0030] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of a product for improving the motor function of elderly individuals is preferred, wherein the product is used to achieve any one or more of the following objectives: (1) increasing muscle mass; (2) enhancing muscle strength; (3) improving motor coordination; (4) improving motor balance; and (5) improving motor endurance.

[0031] The application of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for improving the motor function of elderly individuals, preferably, is that OICR-9429 or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0032] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for improving the motor function of elderly individuals is preferably described in that the effective concentration of OICR-9429 or a pharmaceutically acceptable salt thereof is 3 mg / kg to 30 mg / kg.

[0033] More preferably, the effective concentration of the OICR-9429 or a pharmaceutically acceptable salt thereof is 5 mg / kg.

[0034] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for improving the motor function of older individuals, preferably, the products comprising a medicine.

[0035] More preferably, the drug further includes pharmaceutically acceptable excipients.

[0036] More preferably, the excipients include sulfobutyl ether-β-cyclodextrin.

[0037] More preferably, the dosage form of the drug is a powder, tablet, granule, capsule, solution, emulsion or suspension.

[0038] The application of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for improving the motor function of elderly individuals, preferably, said elderly individuals are mammals, including but not limited to humans, mice, rats, rabbits, dogs, cats and pigs.

[0039] Fourthly, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for the prevention and / or treatment of sepsis, wherein the structural formula of OICR-9429 is as follows:

[0040] .

[0041] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for the prevention and / or treatment of sepsis, preferably, for patients with sepsis including elderly individuals.

[0042] More preferably, the elderly individual is a mammal, including but not limited to humans, mice, rats, rabbits, dogs, cats, and pigs.

[0043] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for the prevention and / or treatment of sepsis, preferably, said sepsis caused by bacterial infection.

[0044] More preferably, the sepsis is caused by infection with Pseudomonas aeruginosa.

[0045] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for the prevention and / or treatment of sepsis is preferred, wherein the OICR-9429 or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0046] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for the prevention and / or treatment of sepsis is preferably carried out at an effective concentration of 3 mg / kg to 30 mg / kg.

[0047] More preferably, the effective concentration of the OICR-9429 or a pharmaceutically acceptable salt thereof is 5 mg / kg.

[0048] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for the prevention and / or treatment of sepsis, preferably, the products comprising a medicament.

[0049] More preferably, the drug further includes pharmaceutically acceptable excipients.

[0050] More preferably, the excipients include sulfobutyl ether-β-cyclodextrin.

[0051] More preferably, the dosage form of the drug is a powder, tablet, granule, capsule, solution, emulsion or suspension.

[0052] Fifthly, the present invention provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for antibacterial infection, wherein the structural formula of OICR-9429 is as follows:

[0053] .

[0054] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for antibacterial infection, preferably, the bacteria include Pseudomonas aeruginosa.

[0055] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for antibacterial infection, preferably, in which the host of the bacterial infection includes an elderly individual.

[0056] More preferably, the elderly individual is a mammal, including but not limited to humans, mice, rats, rabbits, dogs, cats, and pigs.

[0057] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for antibacterial infection is preferred, wherein the OICR-9429 or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0058] According to the present invention, the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for antibacterial infection is preferably carried out at an effective concentration of 3 mg / kg to 30 mg / kg.

[0059] More preferably, the effective concentration of the OICR-9429 or a pharmaceutically acceptable salt thereof is 5 mg / kg.

[0060] The use of OICR-9429 or a pharmaceutically acceptable salt thereof provided by the present invention in the preparation of products for antibacterial infection, preferably, the products comprising a drug.

[0061] More preferably, the drug further includes pharmaceutically acceptable excipients.

[0062] More preferably, the excipients include sulfobutyl ether-β-cyclodextrin.

[0063] More preferably, the dosage form of the drug is a powder, tablet, granule, capsule, solution, emulsion or suspension.

[0064] This invention also provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for treating diseases associated with upregulation of H3K4me3 and / or GSDMD, wherein the structural formula of OICR-9429 is [insert structural formula here].

[0065] .

[0066] This invention also provides the use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of products for reducing H3K4me3 and / or GSDMD expression, wherein the structural formula of OICR-9429 is [insert structural formula here].

[0067] .

[0068] The present invention has the following beneficial effects:

[0069] The OICR-9429 provided by this invention exhibits multiple beneficial technical effects in improving age-related indications. It not only effectively alleviates the inflammatory response caused by pyroptosis during aging but also improves the immune function and anti-infection ability of aging organisms. OICR-9429 also has a significant therapeutic effect on sepsis, inhibiting the rise in inflammation levels caused by infection, improving the survival rate and prognosis of elderly patients. Long-term intervention with OICR-9429 can continuously reduce the chronic inflammatory state of aging organisms, increase muscle mass, enhance motor ability, prolong life, and reduce the occurrence of various age-related diseases. This invention lays the foundation for developing products that comprehensively intervene in aging and related functional decline, and has significant application value in improving the quality of life of the elderly. Attached Figure Description

[0070] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0071] Figure 1 This is a schematic diagram of the OICR-9429 drug administration intervention process provided in Embodiment 1 of the present invention.

[0072] Figure 2 These are the mouse neutrophils provided in Example 1 of this invention. Gsdmd Transcription level detection results.

[0073] Figure 3 This is the statistical result of the protein expression level of GSDMD in neutrophils of mice in each group during the drug administration intervention period provided in Example 1 of the present invention.

[0074] Figure 4 The H3K4me3 in the neutrophils of mice in each group after drug administration intervention provided in Example 1 of this invention is... Gsdmd Statistical results on the enrichment of gene promoter regions.

[0075] Figure 5 This is a statistical analysis of the survival scores of mice in each group after Pseudomonas aeruginosa infection, as provided in Example 2 of this invention.

[0076] Figure 6The results of pyroptosis detection in mice after Pseudomonas aeruginosa infection provided in Example 2 of this invention are shown.

[0077] Figure 7 The results are the serum IL-1β levels of mice in each group after Pseudomonas aeruginosa infection, as provided in Example 2 of this invention.

[0078] Figure 8 These are the pathological examination results of the liver and kidney of mice in each group after Pseudomonas aeruginosa infection, provided in Example 2 of this invention; A is the liver; B is the kidney.

[0079] Figure 9 These are the plasma antigen content detection results of each group of mice provided in Example 3 of the present invention.

[0080] Figure 10 These are the results of detecting the expression levels of inflammatory factors in the liver and kidneys of mice in each group provided in Example 3 of this invention; A represents the kidney; B represents the liver.

[0081] Figure 11 These are the muscle stem cell density detection results of each group of mice provided in Example 3 of the present invention; A is the immunohistochemical fluorescence staining image; B is the statistical result of muscle stem cell density.

[0082] Figure 12 These are the results of the motor ability test of each group of mice provided in Example 3 of the present invention; A is the result of the limb pulling force test; B is the result of the endurance test.

[0083] Figure 13 These are the lifespan tracking results of each group of mice provided in Example 3 of the present invention; A represents the first batch of aged control group and aged OICR-9429 group; B represents the second batch of aged control group and aged OICR-9429 group.

[0084] Figure 14 This is a combined statistical analysis of tissue lesions in two batches of aged control group and aged OICR-9429 group mice provided in Example 3 of the present invention.

[0085] Figure 15 These are the weight tracking results of each group of mice provided in Example 4 of this invention.

[0086] Figure 16 These are the blood biochemical index detection results of each group of mice provided in Example 4 of the present invention; A is blood urea nitrogen; B is creatinine; C is aspartate aminotransferase. Detailed Implementation

[0087] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0088] Unless otherwise specified, the experimental methods involved in the following embodiments are conventional methods in the art. For example, you can refer to the experimental manual in the art or follow the conditions recommended in the manufacturer's instructions.

[0089] Unless otherwise specified, all experimental materials and reagents used in the following examples are commercially available.

[0090] The chemical name of OICR-9429 involved in the following examples is N-[4-(4-Methyl-1-piperazinyl)-3'-(4-morpholinylmethyl)-3-biphenylyl]-6-oxo-4-(trifluormethyl)-1,6-dihydro-3-pyridincarboxamid, CAS number 1801787-56-3, and structural formula is [not specified].

[0091] .

[0092] The sulfobutyl ether-β-cyclodextrin (SBE-β-CD) involved in the following examples is a commonly used excipient for injections, with CAS number 25167-62-8.

[0093] Example 1: Effect of OICR-9429 on GSDMD expression in aged mice

[0094] 1. Experimental Methods

[0095] (1) Laboratory animals

[0096] Eight male C57BL / 6J mice with an initial age of 2 months were used as young mice, and this group was designated as the young control group.

[0097] Male C57BL / 6J mice with an initial age of 17 months were used as naturally aged mice and were randomly divided into two groups of 8 mice each, designated as the aged control group and the aged OICR-9429 group, respectively.

[0098] (2) Drug administration

[0099] After acclimatizing the three groups of mice in this embodiment for 7 days, they were given a one-month drug treatment.

[0100] Preparation of OICR-9429 injection: Use physiological saline containing 10% m / v SBE-β-CD as a solvent, and fully dissolve OICR-9429 in the solvent at a single dose of 5 mg / kg to obtain OICR-9429 injection.

[0101] like Figure 1 As shown, mice in the aged OICR-9429 group were treated with OICR-9429. Each mouse was injected with 200 μL of OICR-9429 injection solution every 3 days via intraperitoneal injection for a total of 10 times until they reached 18 months of age. Subsequent evaluation and analysis were conducted.

[0102] Using the same administration method and frequency, aged control mice were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0103] Using the same administration method and frequency, young control mice were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0104] (3) qPCR detection of GSDMD expression level

[0105] After drug administration, neutrophil samples were collected from the three groups of mice in this example, purified using magnetic beads. RNA was extracted, and cDNA was synthesized via reverse transcription. Using cDNA as a template, the transcription level of GSDMD was detected by qPCR. The detection primers used are shown in Table 1. Two... -ΔΔCt Method calculation Gsdmd The relative expression level, in Gapdh Normalize the internal reference gene.

[0106] Table 1. qPCR primers for Example 1

[0107]

[0108] (4) Flow cytometry detection of GSDMD expression level

[0109] On the day of drug administration, anticoagulation samples were collected from the three groups of mice in this example. Anticoagulation samples were then collected from the aged control group and the OICR-9429 group every week thereafter. Anticoagulation samples were collected from the three groups of mice at the end of drug administration.

[0110] The expression level of GSDMD in the above samples was detected by flow cytometry. The antibody used was GSDMD antibody, manufactured by Abcam, catalog number ab219800.

[0111] (5) ChIP-qPCR detection of H3K4me3 in Gsdmd Enrichment of gene promoter regions

[0112] After drug administration, neutrophils purified by magnetic beads from the three groups of mice in this example were collected, and the H3K4me3 content in the samples was detected by ChIP-qPCR. Gsdmd The enrichment level of the gene promoter region was determined using the following antibodies: H3K4me3 antibody (Abcam, catalog number ab213224) and H3 antibody (Abcam, catalog number ab1791).

[0113] 2. Experimental Results

[0114] like Figure 2 As shown, neutrophils in aged control mice Gsdmd Transcriptional levels were significantly higher in mice than in young control mice; compared with the aged control group, the aged OICR-9429 group mice showed significantly higher transcriptional levels. Gsdmd Transcription levels were significantly reduced, even lower than those in the young control group.

[0115] like Figure 3 As shown, during the one-month treatment period, compared with the aged control group, the expression level of GSDMD protein in mice in the aged OICR-9429 group continued to decrease, eventually reaching a level that of the young control group.

[0116] like Figure 4 As shown, H3K4me3 in neutrophils of aged control mice... Gsdmd The enrichment level of promoter region was significantly higher than that of young control mice; compared with the aged control group, the H3K4me3 enrichment level of aged OICR-9429 mice was significantly decreased.

[0117] The above results indicate that OICR-9429 intervention can reduce the expression level of GSDMD in aged mice and inhibit senescence-induced excessive pyroptosis.

[0118] Example 2: Effect of OICR-9429 on the antiseptic ability of aged mice

[0119] 1. Experimental Methods

[0120] (1) Laboratory animals

[0121] Fifteen male C57BL / 6J mice with an initial age of 2 months were used as young mice, and this group was designated as the young control group.

[0122] Male C57BL / 6J mice with an initial age of 17 months were used as naturally aged mice and randomly divided into two groups: the aged control group (n=25) and the aged OICR-9429 group (n=15).

[0123] (2) Drug administration and construction of sepsis model

[0124] After the three groups of mice in this embodiment were acclimatized for 7 days, they were given a one-month drug treatment and then infected with Pseudomonas aeruginosa (PAO1).

[0125] Preparation of OICR-9429 injection: Use physiological saline containing 10% m / v SBE-β-CD as a solvent, and fully dissolve OICR-9429 in the solvent at a single dose of 5 mg / kg to obtain OICR-9429 injection.

[0126] In the aged OICR-9429 group, each mouse was injected with 200 μL of OICR-9429 injection solution every 3 days via intraperitoneal injection.

[0127] Using the same administration method and frequency, aged control mice were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0128] Using the same administration method and frequency, young control mice were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0129] After the 10th dose, mice in the three groups were administered PAO1 (strain number: ATCC 15692) at a dose of 3 × 10⁻⁶. 6 CFUs is used for infection only, via intraperitoneal injection.

[0130] (3) Immunofluorescence detection of pyroptosis

[0131] After the drug administration was completed, peritoneal lavage fluid samples were collected from the three groups of mice in this example. The expression level of GSDMD was detected by immunofluorescence. The antibodies used were: Ly6G antibody, manufacturer Biolegend, catalog number 127602; and GSDMD antibody, manufacturer Abcam, catalog number ab219800.

[0132] (4) Serum IL-1β level detection

[0133] Serum samples were collected from the three groups of mice in this example 12 hours after infection, and the IL-1β level was detected by ELISA.

[0134] (5) Survival score statistics

[0135] During PAO1 infection, the mortality of the three groups of mice in this example was recorded regularly, and the survival score of each group of mice was calculated.

[0136] (6) Pathological examination

[0137] After the experiment, liver and spleen tissues of mice from the three groups in this example were collected and stained with hematoxylin and eosin (H&E) to analyze the pathological damage of each tissue.

[0138] 2. Experimental Results

[0139] like Figure 5 As shown, in the sepsis model, the survival score of mice in the aged control group was significantly lower than that of mice in the young control group. The survival score of mice in the aged control group had decreased to below 0.5 48 hours after PAO1 infection, and all mice in the aged control group had died by 120 hours after PAO1 infection. Compared with the aged control group, the survival score of aged mice in the aged OICR-9429 group was significantly increased, and remained above 0.7 from 36 hours to 120 hours after PAO1 infection.

[0140] like Figure 6 As shown, in the sepsis model, GSDMD was not expressed in neutrophils of mice in both the young control group and the aged OICR-9429 group, indicating that the pyroptosis level of neutrophils in the aged control group was significantly higher than that in the young control group. However, the pyroptosis level of neutrophils in the aged OICR-9429 group was significantly reduced after OICR-9429 administration, and was comparable to that in the young control group.

[0141] like Figure 7 As shown, in the sepsis model, the serum IL-1β level in mice in the aged control group was significantly higher than that in mice in the young control group; compared with the aged control group, the serum IL-1β level in mice in the aged OICR-9429 group was significantly lower, comparable to that in the young control group.

[0142] like Figure 8 As shown in A and B, in the sepsis model, the tissue damage repair capacity of mice in the young control group was significantly higher than that in the aged control group. The aged control group mice showed severe hepatocyte vacuolar degeneration, blurred edges of spleen white pulp, and granulocyte infiltration in germinal centers 24 to 48 hours after infection. Compared with the aged control group, the tissue damage repair capacity of mice in the aged OICR-9429 group was significantly enhanced.

[0143] The above results indicate that OICR-9429 intervention can inhibit the rise in inflammation levels caused by sepsis, reduce visceral tissue damage, and improve survival rate and prognosis.

[0144] Example 3: Effects of long-term treatment with OICR-9429 on aged mice

[0145] 1. Experimental Methods

[0146] (1) Laboratory animals

[0147] This embodiment uses two batches of mice for the experiment.

[0148] The first batch of mice was divided into two groups: four male C57BL / 6J mice with an initial age of 2 months were designated as young mice, and this group was designated as the young control group. Male C57BL / 6J mice with an initial age of 17 months were designated as naturally aging mice and were randomly divided into two groups of 14 mice each, designated as the aged control group and the aged OICR-9429 group, respectively.

[0149] The second batch of mice was grouped as follows: C57BL / 6J male mice with an initial age of 17 months were used as naturally aged mice and randomly divided into two groups, namely the aged control group (15 mice) and the aged OICR-9429 group (18 mice).

[0150] (2) Drug administration

[0151] Preparation of OICR-9429 injection: Use physiological saline containing 10% m / v SBE-β-CD as a solvent, and fully dissolve OICR-9429 in the solvent at a single dose of 5 mg / kg to obtain OICR-9429 injection.

[0152] For the two batches of aged OICR-9429 groups, the first course of OICR-9429 was administered to mice aged 17-18 months, followed by a second course of OICR-9429 administration at 19-20 months of age. The period from 18-19 months of age was a no-treatment interruption. The procedure for each course was as follows: each mouse was injected with 200 μL of OICR-9429 solution every 3 days for one month via intraperitoneal injection.

[0153] Using the same administration method and frequency, aged control mice were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0154] The young control group in the first batch was not given any medication. When the two groups of older mice in the same batch were finished being given medication, samples were collected from the young control group mice (at this time, they were 2 months old).

[0155] (3) Detection of plasma antigen content

[0156] After the experiment, plasma samples were collected from the elderly control group and the elderly OICR-9429 group mice in this embodiment. The contents of autoantigen dsDNA and Smith were detected by ELISA. Based on the absorbance values ​​measured by the ELISA reader, the contents of each antigen in the elderly OICR-9429 group were normalized to the average value of the elderly control group to obtain the relative contents.

[0157] (4) Detection of tissue inflammatory factor expression levels

[0158] After the experiment, liver and spleen tissues were collected from mice in the young control group, aged control group, and aged OICR-9429 group, respectively. RNA was extracted, and cDNA was synthesized by reverse transcription. Using cDNA as a template, inflammatory factors were detected by qPCR. Il1b , Il6 , Tnf , Ccl2 , Ccl5 , Cxcl1 , Gzmb and Ifng The transcriptional level of [the substance] was measured, and the detection primers used are shown in Table 2. Two [methods] were employed. -ΔΔCt The relative expression levels of each gene were calculated using this method. Gapdh Normalize the internal reference gene.

[0159] Table 2 qPCR primers for Example 3

[0160]

[0161] (5) Muscle stem cell density detection

[0162] After the experiment, quadriceps muscle samples were collected from aged control mice and aged OICR-9429 mice in this embodiment. Immunohistochemistry was used to detect the expression of the muscle stem cell marker PAX7 protein. The antibody used was PAX7 antibody, manufactured by Santa Cruz, catalog number sc-81648. DAPI was used to stain the cell nuclei. Based on the staining results, the density of muscle stem cells was calculated, specifically the percentage of PAX7-positive cells per 100 cell nuclei.

[0163] (6) Sports ability test

[0164] The limb pull strength and rotarod duration of aged mice were tested at 17, 18, 19 and 20 months of age, respectively, in the aged control group and the aged OICR-9429 group.

[0165] (7) Life tracking and tissue lesion detection

[0166] After the experiment, the two batches of aged control mice and aged OICR-9429 mice were kept in the rearing facility until natural death. The mortality rate of each group of mice was recorded regularly, and the cumulative survival rate of each group was calculated. The dead mice were dissected, and the pathological changes in their bodies (such as the presence of tumors, gonadal necrosis, etc.) were observed and recorded.

[0167] 2. Experimental Results

[0168] (1) Inflammation level

[0169] like Figure 9 As shown, compared with the aged control group (the relative contents of dsDNA and Smith were 1.11 and 1.15, respectively), the contents of dsDNA and Smith in the plasma of aged OICR-9429 mice (the relative contents were 0.84 and 0.59, respectively) were significantly reduced, by 24.3% and 48.7%, respectively.

[0170] like Figure 10 As shown in A and B, compared with the young control group, the levels of inflammatory factors in the liver and kidneys of aged mice in the aged control group were significantly increased; while the levels of inflammatory factors in the liver and kidneys of aged mice in the aged OICR-9429 group were significantly lower than those in the aged control group and were basically equivalent to those in the young control group.

[0171] The above results indicate that long-term treatment with OICR-9429 can alleviate chronic inflammation in aged mice, reduce the content of autoantigens in plasma, and decrease the level of inflammation in tissues such as the liver and kidneys.

[0172] (2) athletic ability

[0173] like Figure 11 As shown in A and B, compared with the aged control group (muscle stem cell density of 1.16%), the muscle stem cell density of aged mice in the aged OICR-9429 group (4.81%) was significantly increased, an increase of 315.5%.

[0174] like Figure 12 As shown in A and B, during the treatment period, the limb pulling strength and duration of rotarod movement in aged mice in the aged OICR-9429 group were generally better than those in the aged control group, with increases of 19.2% and 73.8%, respectively.

[0175] The above results indicate that long-term treatment with OICR-9429 can enhance muscle mass and motor function in aged mice.

[0176] (3) Natural death

[0177] like Figure 13 As shown in A and B, among the two batches of aged mice, the cumulative survival probability of the aged OICR-9429 group was significantly higher than that of the aged control group. All mice in the aged control group died naturally before 35 months of age, while more than 30% of the mice in the aged OICR-9429 group (a total of 7 mice) survived until 40 months of age. Compared with the aged control group (median lifespan of 119.9 weeks), the lifespan of the mice in the aged OICR-9429 group (median lifespan of 145.2 weeks) was significantly extended, an increase of more than 15%.

[0178] like Figure 14 As shown, in both batches of aged mice, compared with the aged control group (macro lesion rate of 51.72%), the aged OICR-9429 group mice had significantly reduced macro lesions (macro lesion rate of 9.52%), a reduction of more than 80%.

[0179] The above results indicate that long-term treatment with OICR-9429 can prolong the lifespan of aged mice and reduce the occurrence of age-related diseases.

[0180] Example 4: Biosafety evaluation of OICR-9429

[0181] 1. Experimental Methods

[0182] (1) Laboratory animals

[0183] Two-month-old male C57BL / 6J mice were used as young mice and randomly divided into two groups of four mice each, designated as the young control group and the young OICR-9429 group, respectively.

[0184] Male C57BL / 6J mice with an initial age of 17 months were used as naturally aged mice and were randomly divided into two groups of 12 mice each, designated as the aged control group and the aged OICR-9429 group, respectively.

[0185] (2) Drug administration and weight tracking

[0186] After acclimatizing the four groups of mice in this embodiment for 7 days, they were given a one-month drug treatment.

[0187] Preparation of OICR-9429 injection: Use physiological saline containing 10% m / v SBE-β-CD as a solvent, and fully dissolve OICR-9429 in the solvent at a single dose of 5 mg / kg to obtain OICR-9429 injection.

[0188] In both young and old OICR-9429 mice, 200 μL of OICR-9429 injection solution was injected into each mouse every 3 days for one month via intraperitoneal injection.

[0189] Using the same administration method and frequency, mice in the young control group and the old control group were treated with saline containing 10% m / v SBE-β-CD, with a single administration dose of 200 μL per mouse.

[0190] Record the weight of each mouse before each administration.

[0191] (3) Blood biochemical index detection

[0192] After the experiment, serum samples were collected from mice in the young control group, young OICR-9429 group, old control group, and old OICR-9429 group, respectively. The levels of urea, creatinine, and aspartate aminotransferase (ASAT) were detected by an automated biochemical analyzer to assess the hepatotoxicity and nephrotoxicity of OICR-9429.

[0193] 2. Experimental Results

[0194] (1) Weight changes

[0195] like Figure 15 As shown, during the experiment, there was no significant difference in weight between the young control group and the young OICR-9429 group; there was also no significant difference in weight between the elderly control group and the elderly OICR-9429 group.

[0196] (2) Hepatotoxicity and nephrotoxicity

[0197] like Figure 16 As shown in A to C, there were no significant differences in the levels of urea, creatinine, and aspartate aminotransferase in young mice in the young control group and the young OICR-9429 group. The three blood biochemical indicators of the aged mice in the aged OICR-9429 group were significantly lower than those in the aged control group, and were basically equivalent to those in the young control group and the young OICR-9429 group.

[0198] The above results indicate that long-term treatment with OICR-9429 does not affect the normal growth of either young or old mice; at the same time, long-term treatment with OICR-9429 does not cause liver or kidney toxicity, and even in old mice, long-term treatment with OICR-9429 can alleviate liver and kidney stress, demonstrating excellent biocompatibility.

[0199] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for prolonging lifespan and / or delaying aging in elderly individuals, characterized in that, The structural formula of OICR-9429 is as follows: ； The drug is used to achieve one or more of the following purposes: (1) Improves age-related decline in motor function; (2) Reduce inflammatory response caused by aging.

2. The use of OICR-9429 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving the motor function of elderly individuals, characterized in that, The structural formula of OICR-9429 is as follows: 。 3. The application according to claim 2, characterized in that, The drug is used to achieve one or more of the following purposes: (1) Increase muscle mass; (2) Enhance muscle strength; (3) Improve motor coordination; (4) Improve balance during exercise; (5) Improve exercise endurance.

4. The application according to any one of claims 1 to 3, characterized in that, The effective concentration of OICR-9429 or its pharmaceutically acceptable salt is 3 mg / kg to 30 mg / kg.

5. The application according to claim 4, characterized in that, The effective concentration of OICR-9429 or its pharmaceutically acceptable salt is 5 mg / kg.

6. The application according to any one of claims 1 to 3, characterized in that, The OICR-9429 or its pharmaceutically acceptable salt is the sole active ingredient.

7. The application according to any one of claims 1 to 3, characterized in that, The drug also includes pharmaceutically acceptable excipients.

8. The application according to claim 7, characterized in that, The excipients include sulfobutyl ether-β-cyclodextrin.

Images