Use of alamorelin in the manufacture of a medicament for the treatment of sepsis or sepsis-induced immune dysfunction

Abstract

The application belongs to the technical field of medicine, and particularly relates to application of Alamorelin in preparation of a medicine for treating sepsis or sepsis-induced immune disorders. The Alamorelin is used in preparation of a medicine for inhibiting release of inflammatory factors, promoting thymus regeneration, improving lymphocyte function and metabolism, regulating immune aging process, or relieving organ function damage. The Alamorelin can significantly improve 5-day survival rate of sepsis mice, effectively reduce organ function damage and pathological changes of liver, kidney and the like, significantly reduce serum pro-inflammatory factor (IL-1beta, IL-6, TNF-alpha) level, play a whole-body anti-inflammatory role, obviously relieve thymus atrophy, improve thymus tissue structure and inhibit lymphocyte apoptosis, inhibit expression of a spleen cell aging marker, relieve an immune aging phenotype, and has a significant clinical application potential.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the use of alamolin in the preparation of drugs for treating sepsis or sepsis-induced immune disorders. Background Technology

[0002] Sepsis, a life-threatening organ dysfunction resulting from a dysregulated response to infection, has become the leading cause of death in intensive care units worldwide. Despite significant advancements in early fluid resuscitation, anti-infective therapy, and organ support, the overall mortality rate remains high. The root cause lies in the fact that traditional "one-size-fits-all" treatment strategies, primarily focused on controlling pathogens and maintaining vital signs, have failed to effectively address the persistent and complex host immune system dysfunction caused by sepsis, particularly the progressive depletion and exhaustion of immune function. Current core treatments, such as antibiotic therapy, while targeting pathogens, face limitations including drug resistance, delayed intervention, and the inability to correct immune dysregulation; fluid resuscitation and vasoactive drugs aim to improve circulatory failure but have no direct intervention on the immunopathological process; and organ support such as mechanical ventilation and continuous renal replacement therapy are passive replacements that cannot promote fundamental recovery of immune and organ function. These conventional therapies fail to reverse one of the core pathophysiological aspects of sepsis—immunosuppression and immunosenescence—leading patients, even after surviving the acute phase, to face long-term secondary infections, persistent multi-organ dysfunction, and an increased risk of death.

[0003] Recent studies have revealed that the immunosuppressive state induced by sepsis is highly similar to physiological "immunoresenescence," a condition termed "acute acquired immunosenescence." It is characterized by massive lymphocyte apoptosis and depletion, particularly affecting T cells and B cells, leading to a loss of antigen-specific immune response; myeloid cell dysfunction, manifested as monocyte / macrophage dysfunction or a suppressive phenotype, and impaired antigen-presenting function; failure of immunoregenerative function, with acute atrophy of key lymphoid organs such as the thymus, resulting in the interruption of new T cell output and the inability to rebuild immune system diversity; and the formation of the senescence-associated secretory phenotype (SASP), where aging immune cells secrete large amounts of pro-inflammatory factors and proteases, creating a chronic low-grade inflammatory environment that further damages tissues and suppresses normal immune function. This immunosenescence state is a key mechanism leading to increased susceptibility to nosocomial infections, poor vaccine response, and decreased long-term survival in sepsis patients, making targeted reversal of immunosenescence and restoration of immune homeostasis a promising new strategy in the field of sepsis treatment.

[0004] However, existing research on immunomodulation in sepsis, such as the use of granulocyte colony-stimulating factors, immune checkpoint inhibitors, and interleukins, while having entered clinical trials, has not yet achieved a breakthrough consensus on their efficacy and safety. These immunotherapies generally have limitations: first, they target a single pathway or cytokine, making it difficult to comprehensively correct the complex and networked immune disorders of sepsis; second, their effects are contradictory, as sepsis involves both excessive inflammation and immunosuppression, and a single anti-inflammatory or immunomodulatory strategy may produce opposite harmful effects at different stages or in different patients; third, they lack the dual efficacy of organ protection and immune reconstitution, as most drugs cannot simultaneously achieve the multiple goals of inhibiting excessive inflammation, protecting organ function, and promoting lymphocyte regeneration and functional recovery.

[0005] Therefore, it is particularly important to find a drug that can systematically and multi-targetly regulate the host response, with a particular focus on immune reconstitution. Summary of the Invention

[0006] The purpose of this invention is to provide the application of alamolin in the preparation of drugs for treating sepsis or sepsis-induced immune disorders. The alamolin described in this invention, through activating GHSR receptor-mediated anabolic metabolism and directly regulating the immune system, provides a novel solution for systemic intervention in sepsis-related immune disorders, particularly targeting and reversing the key pathological link of "immunosenescence." This is expected to break through existing treatment bottlenecks and realize a transformation in sepsis treatment from simple life support to promoting the fundamental recovery of immune function.

[0007] The specific implementation process of this invention is as follows: The use of alamolin in the preparation of drugs for the treatment of sepsis or sepsis-induced immune disorders.

[0008] Furthermore, alamolin is used as the sole active ingredient in the preparation of drugs for treating sepsis or sepsis-induced immune disorders.

[0009] Furthermore, alamolin is used in the preparation of drugs that inhibit the release of inflammatory factors, promote thymus regeneration, improve lymphocyte function and metabolism, regulate the immune aging process, or alleviate organ dysfunction.

[0010] Furthermore, the drug aramoline can improve the survival rate of sepsis patients and improve their appetite, weight and / or muscle mass by activating growth hormone secretagogue receptors.

[0011] Furthermore, the drug is a drug that alleviates sepsis-induced organ dysfunction of the liver and kidneys by reducing the elevated ALT and AST levels in the serum caused by sepsis through alamolin.

[0012] Furthermore, the drug achieves a systemic anti-inflammatory effect by reducing the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the serum of sepsis patients through alamolin.

[0013] Furthermore, the drug alleviates thymic atrophy caused by sepsis by aramoline, inhibits thymic lymphocyte apoptosis, and achieves a significant increase in the thymic index and improvement in thymic tissue structure.

[0014] Furthermore, the drug works by inhibiting the expression of β-galactosidase, a marker of splenic cell aging, through alamolin, thereby reducing the accumulation of senescent cells and regulating sepsis-induced immune disorders.

[0015] Furthermore, the alamolin can be used in combination with other treatments, which are selected from antibiotics, vasoactive drugs, immunomodulators, or fluid resuscitation therapy.

[0016] Furthermore, the drug comprises a pharmaceutically effective dose of alamolin or a pharmaceutically acceptable salt, ester, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier; the pharmaceutically acceptable carrier includes one or more excipients, stabilizers, solubilizers, pH adjusters, isotonic agents, or sustained-release materials; the dosage form of the drug is an oral preparation, an injection, or a transdermal preparation, wherein the oral preparation is preferably a tablet, capsule, or oral liquid.

[0017] The positive effects of this invention: (1) Existing research indicates that alamolin, as a ghrelin analogue, is primarily used to improve appetite and weight in patients with cancer cachexia. This invention is the first to propose and demonstrate that alamolin can systematically intervene in and reverse sepsis-induced acute acquired immunosenescence through its unique dual-action effect (activation of GHSR receptor-mediated anabolic metabolism and immune regulation). This transcends the traditional treatment concept that focuses solely on pathogen clearance and organ support, shifting the therapeutic target to the fundamental reconstruction of the immune system, providing a new approach and direction for overcoming the clinical challenge of high mortality and long-term sequelae of sepsis.

[0018] (2) This invention protects the specific effects and targets of alamolin in reversing immunosenescence. This invention does not propose a general concept of immune regulation, but rather clarifies the key aspects of alamolin's action on immunosenescence through research: promoting thymus regeneration to restore immune regeneration capacity, improving lymphocyte function and metabolism, and alleviating age-associated secretory phenotype (SASP)-mediated chronic inflammation. In particular, its regenerative effect on the thymus, the "engine" of the immune system, directly targets the core defect of sepsis that prevents the immune system from self-repairing. This effect provides a clear and actionable molecular and cellular target for the field of sepsis immunotherapy.

[0019] (3) This invention uses a more clinically relevant severe sepsis model for validation, making the conclusions more valuable for translation. Many preclinical studies on immunomodulatory therapy for sepsis currently use acute inflammatory models such as LPS injection, which have a simple pathological process and are difficult to simulate the complex, multi-stage immune dysregulation process of clinical sepsis. The validation of this invention is based on the cecal ligation and perforation (CLP) model, which better simulates the real pathological process of clinical multimicrobial infection, bacterial translocation, and progressive organ damage, especially effectively inducing significant thymic atrophy and lymphocyte depletion, among other immunosenescent phenotypes. Demonstrating the effectiveness of alamolin on this model greatly enhances the persuasiveness and feasibility of translating this treatment strategy into clinical practice.

[0020] (4) This invention has the potential advantage of "multi-effect synergy and treating both the symptoms and the root cause". Alamorin, on the one hand, improves the overall wasting state of sepsis patients by promoting anabolism, providing a material basis for immune recovery; on the other hand, it directly regulates the immune aging process and promotes immune reconstitution. This dual effect of simultaneously improving the overall condition and repairing the immune system is not possessed by existing single-target immune agonists or inhibitors. It is expected to control acute inflammation while preventing or reducing the risk of long-term immunosuppression and secondary infection, achieving benefits throughout the entire process from acute treatment to long-term prognosis improvement.

[0021] (5) This invention has a known clinical safety profile and a relatively low risk of drug translation. Compared to completely novel compounds, alamolin has accumulated a large amount of clinical safety data in cancer cachexia patients. This invention opens up a completely new indication for it (sepsis and sepsis-related immune disorders), which allows its subsequent clinical development to be based on a relatively well-defined drug safety profile, significantly reducing the unknown risks of translating from laboratory to clinical application and accelerating its development as a new strategy for the treatment of sepsis.

[0022] (6) This invention not only proposes a novel use of alamolin for the treatment of sepsis and its induced immune aging, but also demonstrates significant innovation and practicality in terms of mechanism depth, model relevance and clinical translation prospects, providing a strong candidate and scientific basis for the development of next-generation sepsis treatment drugs. Attached Figure Description

[0023] Figure 1 A graph showing that alamolin significantly improves the 5-day survival rate of septic mice; Figure 2 The protective effect of alamolin on organ function in septic mice is shown in Figures A and B, which represent serum liver function markers (ALT and AST). Figures C and D show the H&E staining results of liver and kidney tissue pathological sections, respectively. Figure 3A graph showing that alamolin significantly downregulated the serum levels of inflammatory factors IL-1β, IL-6, and TNF-α in septic mice. The data are expressed as mean ± standard error (SEM) and are derived from three independent replicate experiments. Figure 4 To illustrate the effect of alamolin on thymic atrophy in septic mice, Figures A and B show the changes in thymic index at 24 and 48 hours post-surgery; Figures C and D show the pathological morphology of thymic tissue stained with H&E at 48 hours post-surgery. Figure 5 Image showing β-galactosidase staining in mouse spleen tissue. Detailed Implementation

[0024] The technical solution of the present invention will be described in more detail below with reference to experiments.

[0025] Unless otherwise stated, the terms used herein have the meanings commonly understood by those skilled in the art. Unless otherwise specified, all experimental materials and reagents used in the experiments are commercially available. For experiments where specific techniques or conditions are not specified, the procedures described in the literature in this field or the product instructions should be followed.

[0026] Alamolin, an orally administered, bioactive ghrelin analogue, exhibits unique potential advantages in treating sepsis. It possesses a dual mechanism of action: firstly, by activating the growth hormone secretagogue receptor (GHSR), it effectively promotes appetite, increases lean body mass, and reverses the severe wasting state associated with sepsis, providing a material basis for immune recovery; secondly, its downstream signaling pathways (such as the growth hormone / insulin-like growth factor-1 axis) have been shown to have direct regulatory effects on the immune system. Regarding its potential role in reversing immunosenescence, alamolin promotes thymic regeneration—the GH / IGF-1 axis is a known important positive regulator of thymic structure and function, promoting thymic epithelial cell proliferation, reducing thymic cell apoptosis, and enhancing thymic output function, thereby directly counteracting sepsis-induced acute thymic atrophy; simultaneously, it improves lymphocyte function, enhances T cell proliferation and survival, improves immune cell metabolism, and regulates the cytokine spectrum towards a more balanced state; furthermore, it alleviates age-related chronic inflammation by regulating SASP. More importantly, alamolin has undergone extensive clinical studies in patients with cancer cachexia, demonstrating an acceptable safety profile, which provides preliminary safety data to support its application in the preparation of drugs for treating sepsis. With its unique anabolic and potential immunomodulatory properties, alamolin offers a highly innovative and promising new use for sepsis and its resulting long-term immune sequelae.

[0027] In practical production applications, alamorelin can be used as the main active ingredient, formulated into pharmaceutically acceptable salts, esters, solvates, or prodrugs, for the preparation of drugs to treat sepsis or sepsis-induced immune disorders.

[0028] Example 1 (i) In this embodiment, alamolin powder was used and purchased from the Selleck reagent website; the pH value of PBS phosphate buffer was 7.2-7.4.

[0029] The method for preparing the Alamolin working solution in this embodiment is as follows: 20 mg of alamolin powder was dissolved in 100 μL of dimethyl sulfoxide (DMSO) to prepare a clear stock solution, which was then added to 800 μL of polyethylene glycol 300 (PEG300) and mixed thoroughly until clear. 100 μL of Tween 80 was added to the above system and mixed thoroughly until clear. PBS phosphate buffer was then added to bring the volume to 10 mL and mixed thoroughly to obtain a 2 mg / mL alamolin working solution. The alamolin working solution should be prepared and used immediately.

[0030] The placebo solvent (Vehicle) used in the control group was prepared by adding equal amounts of DMSO, PEG300, and Tween80 to the working solution, and then adding PBS solution to bring the volume to 10 mL and mixing thoroughly.

[0031] (ii) Animal feeding and model making (1) The animal source used in this embodiment is: SPF grade 6-8 week male C57BL / 6 mice with an average weight of 20-25g, provided by the Experimental Animal Center of Xi'an Jiaotong University.

[0032] (2) Animal rearing process: Adult male healthy C57BL / 6 mice were provided by the Experimental Animal Center of Xi'an Jiaotong University. The mice were placed in a rearing room under strict conditions of 12-hour photocycle, 22±2℃ and 70% humidity for one week of acclimatization feeding, during which the mice had free access to water and food. After 7 days, the mice were randomly divided into three groups: sham-operated group (Sham-Vehicle), sepsis model group (CLP-Vehicle), and alamorin treatment group (CLP-Ana 30mg / kg), with 40 mice in each group. The alamorin treatment group was given intraperitoneal injection of alamorin 30mg / kg daily 2 hours after the establishment of the sepsis model. Mice in the sham-operated group and the sepsis model group were given the same volume of Vehicle.

[0033] (3) Modeling (CLP model): Cecal ligation and perforation (CLP) A mouse sepsis model was established using cecal ligation and perforation (CLP).

[0034] (3.1) Prepare a sodium pentobarbital solution (2g of sodium pentobarbital is dissolved in 100mL of water for injection to prepare a 2.0% solution by mass / volume ratio) and administer it intraperitoneally to mice at a dose of 40 mg / kg. After anesthetizing the mice, fix them in a supine position on the operating table, and use an electric shaver to remove the fur from their abdomens to thoroughly clean the surgical area. Perform three alternating circumferential disinfections with povidone-iodine and 75% alcohol by volume.

[0035] (3.2) Make a longitudinal skin incision of about 1-2 cm along the midline of the lower abdomen, bluntly dissect the subcutaneous tissue, and expose the linea alba. Gently lift the linea alba with forceps and make a small incision of about 0.8-1 cm with fine scissors, taking care to avoid damaging internal organs. Gently bring the cecum out of the abdominal cavity with blunt forceps or a cotton swab moistened with physiological saline, and ligate it at 1 / 3 of the ileocecal valve with 4-0 silk suture. The ligation should be firm to block blood flow, but the intestinal segment should not be severed. After ligation, the distal cecum should immediately show ischemia and darken in color. Use an 18G needle to penetrate the intestinal wall on both sides of the ligated segment of the cecum, creating an "in-out" perforation. Gently squeeze the ligated segment of the cecum to allow a small fecal pellet (about the size of a grain of rice) to overflow from the perforation into the abdominal cavity to ensure the establishment of a source of infection. Take care to avoid excessive squeezing, which may lead to large-area contamination or tearing of the perforation. Using forceps, the cecum, along with the small amount of spilled feces, was gently returned to the abdominal cavity. Finally, the abdominal incision was sutured layer by layer with 4-0 sterile non-absorbable sutures, and the area was disinfected.

[0036] (3.3) In the sham surgery group, no ligation or cecal perforation was performed. After disinfection, the abdominal cavity was opened, the cecum was located and placed back into the abdominal cavity for incision suturing. In the sepsis model group and the alamolin treatment group, complete disinfection, ligation, cecal perforation and incision suturing were performed.

[0037] (3.4) After the surgery, fluid resuscitation was performed. All mice were injected with 0.5 mL of physiological saline to replenish fluid loss. The mice were placed individually in warm cages (37°C heat pads or heat lamps) with clean bedding until they were fully awake.

[0038] (3.5) Provide food and water to the mice after they have recovered. The mice that have successfully developed the model showed symptoms such as delayed recovery, lethargy, sluggishness, reduced food and water intake, rapid breathing, curled-up body, erect hair, loose stools, and sluggish response to external stimuli. Bloody exudate, intestinal edema and adhesion, and necrosis and blackening of the cecum were also observed after abdominal dissection.

[0039] (iii) Alamolin can improve the 5-day survival rate of septic mice. Experimental Procedure: After surgery, all mice were placed in warm, clean cages with free access to food and water. The alamorin treatment group (CLP-Ana 30 mg / kg) received intraperitoneal injections of alamorin (30 mg / kg) once daily, while the sham-operated group (Sham-Vehicle) and the sepsis model group (CLP-Vehicle) received an equal volume of vehicle. The primary endpoint was 5-day postoperative survival rate, with the number of surviving mice in each group recorded every 12 hours, and observation continued until 120 hours postoperatively.

[0040] Experimental results: like Figure 1 Survival curves show that all mice in the Sham-Vehicle group survived, with a 5-day survival rate of 100%. The survival rate of mice in the CLP-Vehicle group was significantly reduced, with a 5-day survival rate of 30.6%. The alamolin treatment group (CLP-Ana 30 mg / kg) significantly improved the survival rate of septic mice, with a 5-day survival rate of 61.1%, which was significantly different from that of the CLP-Vehicle group (P < 0.01).

[0041] Experimental conclusion: In the sepsis mouse model (CLP model) used in this invention, intraperitoneal injection of alamolin (30 mg / kg / day) significantly improved the 5-day survival rate of mice, suggesting that alamolin has positive therapeutic potential for sepsis.

[0042] (iv) Alamolin can alleviate organ dysfunction in septic mice. 1. Experimental Results 1.1 Effects of Alamolin on Liver Function Indicators in Sepsis-Induced Mice The degree of liver function damage in mice of each group was assessed 24 and 48 hours after surgery by detecting serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Results are as follows... Figure 2 As shown in A and B.

[0043] 24 hours after surgery Figure 2 Compared with the Sham-Vehicle group, the serum ALT and AST levels in mice in the CLP-Vehicle group were significantly increased (P < 0.01). In the alamolin treatment group (CLP-Ana, 30 mg / kg), the above indicators were significantly lower than those in the CLP-Vehicle group (P < 0.01), indicating that alamolin can effectively alleviate acute liver injury in the early stage (24 hours) of sepsis.

[0044] 48 hours after surgery Figure 2(B) Liver injury markers remained at high levels in the CLP-Vehicle group. The ALT and AST levels in the alamolin treatment group (CLP-Ana, 30 mg / kg) were significantly lower than those in the CLP-Vehicle group (ALT P < 0.05, AST P < 0.01), indicating that continuous two-day administration of alamolin could persistently alleviate liver injury in septic mice.

[0045] 1.2 Effects of Alamolin on the histopathology of major organs in septic mice like Figure 2 As shown in C and D, the pathological changes in liver and kidney tissues 24 hours after surgery were observed by hematoxylin-eosin (HE) staining.

[0046] Liver tissue: The Sham-Vehicle group showed clear liver lobule structure and neatly arranged hepatocytes; the CLP-Vehicle group showed obvious pathological damage such as inflammatory cell infiltration, hepatocyte edema and punctate necrosis; the alamolin treatment group (CLP-Ana, 30mg / kg) showed significant reduction of the above lesions, and significant improvement in liver tissue structure destruction and inflammatory infiltration.

[0047] Kidney tissue: In the Sham-Vehicle group, the glomerular and tubular structures were normal; in the CLP-Vehicle group, glomerular congestion, tubular epithelial cell edema and vacuolar degeneration were observed, and casts were seen in some tubular lumens; in the alamorin treatment group (CLP-Ana, 30 mg / kg), kidney tissue damage was significantly reduced, the tubular structure remained basically intact, and cast formation was reduced.

[0048] 2. Experimental Conclusions The results of this experiment indicate that intraperitoneal injection of alamolin (30 mg / kg, once daily) significantly reduced elevated ALT and AST levels in the serum of septic mice, effectively alleviating liver function damage. Furthermore, histopathological observation further confirmed that alamolin treatment significantly reduced histopathological damage to the liver and kidneys of septic mice. These biochemical and morphological results collectively demonstrate that alamolin has a clear protective effect against sepsis-induced multi-organ dysfunction.

[0049] (v) Systemic anti-inflammatory effects of alamolin on septic mice 1. Experimental Results 1.1 Effects of Alamolin on Systemic Inflammatory Factor Levels in Septic Mice in the Early Stage (24 Hours Post-Surgery) like Figure 3 As shown in Figure A, the concentrations of key pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the serum of mice in each group 24 hours after surgery were detected by enzyme-linked immunosorbent assay (ELISA).

[0050] Compared with the Sham-Vehicle group, the serum levels of IL-1β, IL-6, and TNF-α in mice in the CLP-Vehicle group were significantly increased (P < 0.001), indicating that sepsis induced a strong systemic inflammatory response. In contrast, the levels of these inflammatory factors in the alamolin treatment group (CLP-Ana, 30 mg / kg) were significantly lower than those in the CLP-Vehicle group (P < 0.01), suggesting that alamolin can effectively inhibit the excessive release of pro-inflammatory factors in the early stages of sepsis.

[0051] 1.2 Effects of Alamolin on Systemic Inflammatory Factor Levels in Septic Mice in the Late Stage (48 Hours Post-Surgery) Postoperative inflammatory factor test results 48 hours later Figure 3 As shown in B, the levels of IL-1β, IL-6, and TNF-α in the CLP-Vehicle group were still significantly higher than those in the Sham-Vehicle group (P < 0.01).

[0052] The anti-inflammatory effect of alamolin remained significant and sustained at this phase point. Compared with the CLP-Vehicle group, the serum levels of IL-1β, IL-6, and TNF-α in the alamolin treatment group (CLP-Ana, 30 mg / kg) were further reduced, all significantly lower than those in the CLP-Vehicle group (P < 0.001), demonstrating a strong and sustained systemic anti-inflammatory effect.

[0053] 2. Experimental Conclusions The results of this experiment indicate that intraperitoneal injection of alamolin significantly reduces the serum levels of IL-1β, IL-6, and TNF-α in septic mice. This inhibitory effect appeared as early as 24 hours post-surgery and persisted until 48 hours post-surgery. These results confirm, at the level of inflammatory mediators, that alamolin has a clear systemic anti-inflammatory effect on septic mice, which is one of the important mechanisms by which alamolin improves survival rate and protects organ function.

[0054] (vi) The alleviating effect of alamolin on thymic atrophy in septic mice 1. Experimental Results 1.1 Effects of Alamorin on Thymus Weight and Thymus Index in Septic Mice The thymus index (thymus wet weight / body weight) is a classic indicator for assessing the degree of thymus atrophy. For example... Figure 4 As shown in A and B, the thymus status of mice in each group was assessed 24 and 48 hours after surgery through gross observation and quantitative analysis.

[0055] 24 hours after surgery Figure 4 A): At 24 hours post-surgery, compared with the Sham-Vehicle group, the CLP-Vehicle group showed significant thymic atrophy and a significantly lower thymic index (P < 0.001). The thymic index in the alamolin treatment group (CLP-Ana, 30 mg / kg) was significantly higher than that in the CLP-Vehicle group (P < 0.05), indicating that alamolin can effectively slow the progression of thymic atrophy in the early stages of sepsis.

[0056] 48 hours after surgery Figure 4 B): Thymic atrophy was further exacerbated in the CLP-Vehicle group, with a significant reduction in thymic volume and a thymic index lower than 24 hours post-surgery, showing a more significant difference compared to the Sham-Vehicle group. In contrast, the thymic index significantly increased in the alamorin treatment group (CLP-Ana, 30 mg / kg) (P < 0.01), indicating that alamorin can sustainably alleviate sepsis-induced thymic atrophy.

[0057] 1.2 Effects of Alamolin on the Pathological Structure of Thymus Tissue in Septic Mice like Figure 4 As shown in C and D, the histopathological changes of the thymus were further observed 48 hours after surgery by hematoxylin-eosin (HE) staining.

[0058] Under low magnification ( Figure 4 C): The Sham-Vehicle group had an intact thymus structure, clear corticomedullary boundary, and high cell density; the CLP-Vehicle group showed typical severe atrophy, with reduced thymus volume, blurred corticomedullary structure, and significantly reduced cell density; the alamorin treatment group (CLP-Ana, 30 mg / kg) had better preservation of thymus tissue structure, reduced atrophy, and significantly higher cell density than the CLP-Vehicle group.

[0059] Under high magnification ( Figure 4 D): In the Sham-Vehicle group, the lymphocytes in the thymic cortex were tightly arranged, and clear thymic bodies were visible in the medulla. In the CLP-Vehicle group, the corticomedullary boundary was unclear, the structure was disordered, and a large number of scattered "starry sky" apoptotic cell fragments were visible. In the alamolin treatment group (CLP-Ana, 30 mg / kg), the above pathological changes were significantly improved, the apoptotic cell fragments were significantly reduced, and the corticomedullary structure was partially restored.

[0060] 2. Experimental Conclusions The results of this experiment indicate that intraperitoneal injection of alamolin (30 mg / kg, once daily) effectively alleviates thymic atrophy in septic mice, mainly manifested by a significant increase in the thymic index, improved thymic tissue structure, and significant inhibition of thymic lymphocyte apoptosis. This suggests that alamolin has a protective effect against sepsis-related immune organ damage and can help improve the immunosuppressive state in the later stages of sepsis.

[0061] (vii) Effects of alamolin on senescence-related phenotypes of spleen cells in septic mice 1. Experimental Results Cellular senescence is a key marker of tissue dysfunction and immunosenescence, and senescence-associated β-galactosidase (SA-β-gal) activity is a commonly used biological marker for identifying senescent cells. Figure 5 (Results of SA-β-gal staining in spleen tissue) As shown, the changes in SA-β-gal activity in spleen tissue of mice in each group 48 hours after surgery were detected by histochemical staining to assess the degree of sepsis-induced immune cell senescence and the intervention effect of alamolin.

[0062] Compared with the Sham-Vehicle group, the CLP-Vehicle group showed a significant increase in SA-β-gal-positive areas and a marked increase in the number of positive cells (appearing blue-green) in the spleen tissue, indicating that sepsis can induce significant cellular senescence in spleen tissue. This suggests that sepsis, in addition to causing acute inflammation and organ damage, can also accelerate the aging process of immune organs.

[0063] Compared with the CLP-Vehicle group, the aramoline treatment group (CLP-Ana, 30 mg / kg) showed a significant reduction in the number of SA-β-gal positive areas in the spleen, a significantly weaker positive intensity, and staining results closer to those of the Sham group. This indicates that aramoline can effectively inhibit the cellular senescence process in the spleen tissue of septic mice.

[0064] 2. Experimental Conclusions The results of this experiment indicate that intraperitoneal injection of alamolin (30 mg / kg) significantly reduced the activity of senescence-associated β-galactosidase (SA-β-gal) in the spleen tissue of septic mice, and decreased the accumulation of senescent cells. This suggests, from the perspective of cellular senescence, that alamolin can alleviate the sepsis-induced senescent phenotype of immune organs and can regulate sepsis-related immune dysfunction by inhibiting premature senescence of immune cells, providing a new direction for exploring mechanisms to improve the long-term prognosis of sepsis.

[0065] This invention, through a series of animal experiments, confirms that alamolin has clear and multiple beneficial effects on sepsis. Specifically, it significantly improves the 5-day survival rate of septic mice; effectively reduces functional damage and pathological changes in organs such as the liver and kidneys; significantly reduces serum levels of pro-inflammatory factors (IL-1β, IL-6, TNF-α), exerting a systemic anti-inflammatory effect; significantly alleviates thymic atrophy, improves thymic tissue structure, and inhibits lymphocyte apoptosis; and inhibits the expression of splenic cell senescence markers, alleviating the immunosenescence phenotype. These results indicate that alamolin can improve the prognosis of sepsis through multiple effects, including anti-inflammation, organ protection, and immunomodulation, and has significant potential for clinical application.

[0066] The above are merely preferred embodiments of the present invention and are not intended to limit the scope of the invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of alamolin in the preparation of drugs for the treatment of sepsis or sepsis-induced immune disorders.

2. The application according to claim 1, characterized in that: The use of alamolin as the sole active ingredient in the preparation of drugs for the treatment of sepsis or sepsis-induced immune disorders.

3. The application according to claim 1, characterized in that: The application of alamolin in the preparation of drugs that inhibit the release of inflammatory factors, promote thymus regeneration, improve lymphocyte function and metabolism, regulate the immune aging process, or alleviate organ dysfunction.

4. The application according to claim 1, characterized in that: Alamolin, one of the drugs in question, can improve the survival rate of sepsis patients and improve their appetite, weight, and / or muscle mass by activating growth hormone secretagogue receptors.

5. The application according to claim 1, characterized in that: The drug is a medication that alleviates sepsis-induced organ dysfunction of the liver and kidneys by reducing elevated ALT and AST levels in the serum caused by sepsis.

6. The application according to claim 1, characterized in that: The drug works by reducing the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the serum of sepsis patients, thereby achieving a systemic anti-inflammatory effect.

7. The application according to claim 1, characterized in that: The drug works by relieving thymic atrophy caused by sepsis and inhibiting thymic lymphocyte apoptosis, thereby significantly increasing the thymic index and improving the thymic tissue structure.

8. The application according to claim 1, characterized in that: The drug works by inhibiting the expression of β-galactosidase, a marker of senescence in spleen cells, through alamolin, thereby reducing the accumulation of senescent cells and regulating sepsis-induced immune disorders.

9. The application according to claim 1, characterized in that: The alamolin can be used in combination with other treatments, which are selected from antibiotics, vasoactive drugs, immunomodulators, or fluid resuscitation therapy.

10. The application according to claim 1, characterized in that: The drug comprises a pharmaceutically effective dose of alamolin or a pharmaceutically acceptable salt, ester, solvate or prodrug thereof, and a pharmaceutically acceptable carrier; the pharmaceutically acceptable carrier includes one or more excipients, stabilizers, solubilizers, pH adjusters, isotonic agents or sustained-release materials; the dosage form of the drug is an oral preparation, an injection or a transdermal preparation, wherein the oral preparation is preferably a tablet, capsule or oral liquid.

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