Application of ester compound in preparation of sepsis prevention and treatment medicine

Abstract

The invention discloses an application of an ester compound in preparation of a sepsis prevention and treatment medicine. The structural formula of the ester compound is as shown in the following formula: # imgabs0 #. The compound can relieve multi-organ injury caused by sepsis, relieve organ dysfunction caused by sepsis and reduce release of inflammatory factors and injury factors, and can be used for preventing, relieving and / or treating sepsis and other related diseases and improving sepsis organ injury and inflammatory response.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to the application of an ester compound in the preparation of drugs for preventing and treating sepsis. Background Art

[0002] Sepsis is life-threatening organ dysfunction caused by the dysregulation of the body's response to infection. It is the final common pathway of death in severe infectious diseases. In the early stage of infection, pathogenic microorganisms and their toxins enter the systemic circulation from the local area, activate the host immune system, cause a large release of inflammatory factors, trigger cytokine storms, lead to multiple organ dysfunction, and ultimately cause the death of the host. The global incidence of sepsis is approximately 48.9 million, and the number of sepsis-related deaths is approximately 11 million, accounting for about 20% of the total global deaths. Sepsis is a complex disease process involving multiple organs. A series of physiological changes caused by infection may lead to dysfunction of organs such as the lungs, kidneys, liver, and intestines. Treating sepsis requires taking effective measures as soon as possible to minimize the occurrence of organ failure. Through timely and accurate intervention, the risk of organ failure can be reduced, and the survival rate of patients can be improved. Although the understanding of the pathogenesis of sepsis and treatment methods have been continuously improved in recent years, the in-hospital mortality rate of sepsis patients remains high, and there is an urgent need to develop new drugs to effectively prevent or treat sepsis. Summary of the Invention

[0003] One object of the present invention is to provide the application of a compound represented by the following formula in the preparation of drugs for preventing and treating sepsis,

[0004]

[0005] The said application includes: alleviating organ damage caused by sepsis, including sepsis-induced lung injury, sepsis-induced intestinal injury, sepsis-induced liver injury, and sepsis-induced kidney injury; alleviating organ dysfunction caused by sepsis; increasing the survival rate or inhibiting body temperature decline; improving histopathological damage of the lungs, intestines, liver, and kidneys; reducing the expression of serum inflammatory factors interferon-β (IFN-β), interleukin-1β (IL-1β), and chemokine (CXCL1); reducing the expression of damage-related molecules C-reactive protein (CRP), creatine kinase (CK), alanine aminotransferase (ALT), and blood urea nitrogen (BUN).

[0006] Another object of the present invention is to provide the application of a pharmaceutical composition in the preparation of drugs for preventing and treating sepsis. The pharmaceutical composition includes an active ingredient, and the active ingredient is a compound represented by the following formula:

[0007]

[0008] In the present invention, the administration route applied can be oral or injection, specifically intraperitoneal, subcutaneous, intravenous or intramuscular injection; pharmaceutically acceptable carriers such as diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. can also be added to prepare a pharmaceutical composition in a certain form, such as powders, pills, capsules, tablets, microcapsules, soft capsules, membranes, suppositories, injections, ointments, powders, granules, aerosols and other preparations.

[0009] Studies have shown that the compound can effectively inhibit the systemic inflammatory response caused by cytokine storm by significantly reducing the levels of pro-inflammatory factors (such as IL-1β, IFN-β, CXCL1) in the serum of septic mice. In addition, the compound can improve the phenomenon of body temperature reduction in septic mice and show significant effects in protecting the pathological damage of multiple organs such as the liver, kidney, lung and intestine. Experimental data further prove that the efficacy of the compound is dose-dependent and significantly improves the survival rate of mice within 72 hours at a high dose (3 mg / kg). The present invention reveals the potential application value of the compound in regulating the pathological process of sepsis in multiple aspects.

[0010] Compared with the prior art, the present invention has the following remarkable advantages:

[0011] The present invention for the first time confirms that the ester compound can be used for preventing, alleviating and / or treating sepsis and other related diseases, and improving organ damage and inflammatory response in sepsis. Brief Description of the Drawings

[0012] Figure 1 Effects of Compound No. 6 at different doses on the survival rate and body temperature of CLP mice, where A is the statistical chart of the survival rate of CLP mice within 72 hours after surgery; B is the chart of the body temperature change of CLP mice within 72 hours after surgery;

[0013] Figure 2 Effects of Compound No. 6 at different doses on the pathological damage of the lungs, intestines, livers and kidneys of septic mice induced by CLP surgery, where A is the HE staining diagram of the liver, kidney, lung and intestine, and B is the statistical chart of the HE staining pathological score (n = 6, expressed as mean ± SD, ## P < 0.01 compared with the sham operation group, * P < 0.05 and ** P < 0.01 compared with the model group, scale bar = 100 μm);

[0014] Figure 3Effects of Compound 6 at different doses on the levels of serum inflammatory factors in septic mice induced by CLP surgery. Among them, A is the statistical chart of interferon-β (IFN-β), B is the statistical chart of interleukin-1β (IL-1β), and C is the statistical chart of chemokine (CXCL1) (n = 8, expressed as mean ± SD, ## P < 0.01 compared with the sham operation group, * P < 0.05 and ** P < 0.01 compared with the model group);

[0015] Figure 4 Effects of Compound 6 at different doses on the levels of damage-related factors in the serum of CLP mice. Among them, A is the statistical chart of C-reactive protein (CRP), B is the statistical chart of creatine kinase (CK), C is the statistical chart of alanine aminotransferase (ALT), and D is the statistical chart of blood urea nitrogen (BUN) (n = 8, expressed as mean ± SD, ## P < 0.01 compared with the sham operation group, ** P < 0.01 compared with the model group). Detailed implementation manners

[0016] Aiming at the complex pathological mechanisms of sepsis and the deficiencies of existing treatment methods, the present invention aims to explore and verify the new use of an ester compound (Compound 6) in the prevention and treatment of sepsis, and provide an innovative and efficient solution. Through the research on the mechanism of action and pharmacological effects of this compound, the present invention discovers for the first time that Compound 6 can regulate the cytokine storm by reducing the levels of pro-inflammatory factors (such as IL-1β, IFN-β, CXCL1), improve the multi-organ pathological damage caused by the inflammatory response, and particularly show excellent protective effects in liver, kidney, lung and intestinal tissues. In addition, Compound 6 can also effectively improve the survival rate of septic mice and inhibit the decrease in body temperature, showing a dose-dependent therapeutic effect. The present invention verifies the efficacy of Compound 6 in sepsis from multiple aspects, covering multiple levels such as inflammation inhibition, organ protection and survival rate improvement, providing a scientific basis for subsequent drug development and clinical application.

[0017] The technical solution of the present invention takes Compound 6 ester as the core and effectively controls the key pathological processes of sepsis through multiple mechanisms. The present invention discovers for the first time that Compound 6 can inhibit the abnormal release of inflammatory factors, improve the pathological damage of multiple organs and significantly improve the survival rate, demonstrating its significant advantages as a therapeutic drug for sepsis.

[0018] Through experimental research, the present invention discovers that Compound 6 can significantly reduce the levels of pro-inflammatory factors such as IL-1β, IFN-β, and CXCL1 in the serum in a cecal ligation and puncture (CLP)-induced mouse sepsis model. The massive release of inflammatory factors is a key lethal factor in the course of sepsis. Compound 6 controls the pathological progression of sepsis by regulating inflammatory factors and inhibiting the systemic inflammatory response caused by cytokine storm. In addition, this compound can also improve the problem of body temperature decline in septic mice, showing a significant dose-dependent effect. In the experiment, when the dose reaches 3 mg / kg, its effect is better than that of the positive control drug, showing a strong anti-inflammatory effect.

[0019] In terms of multi-organ protection, Compound 6 shows excellent curative effects. Through histopathological detection, it is found that this compound can significantly reduce the tissue damage of the liver, kidney, lung, and intestine in septic mice, reduce the pathological score, and reduce tissue edema and vascular leakage. When the dose is 3 mg / kg, its multi-organ protection effect is the most significant, better than that of the traditional drug dexamethasone. This compound not only improves the tissue damage caused by acute inflammatory response but also plays an important role in maintaining the functional integrity of multiple organs, fully demonstrating its potential for treating sepsis.

[0020] Compound 6 described in the present invention can be administered by oral or injection methods, including intraperitoneal, subcutaneous, intravenous, or intramuscular injection. In addition, Compound 6 can be formulated into various dosage forms in combination with excipients, fillers, and other auxiliary materials, including tablets, capsules, injections, aerosols, etc., to meet different clinical needs.

[0021] The preferred embodiments of the present invention will be described in detail below in conjunction with the examples. It should be understood that the following examples are given only for the purpose of illustration and are not used to limit the scope of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0022] The experimental methods used in the following examples are all conventional methods unless otherwise specified.

[0023] The materials, reagents, etc. used in the following examples can be obtained from commercial channels unless otherwise specified.

[0024] Example 1

[0025] Effects of Different Doses of Compound 6 on the Mortality and Body Temperature of Septic Mice Induced by CLP Surgery

[0026] I. Experimental Materials

[0027] 1. Experimental Animals

[0028] Male C57BL / 6J mice, weighing 22 - 24 g, of SPF grade, were purchased from Nanjing Annokang Biotechnology Co., Ltd., meeting the quality standards of ordinary experimental animals. License number: SCXK (Jiangsu) 2020 - 0009. The mice were housed separately in cages, with free access to water and food. The temperature of the breeding room was 23 - 25 °C, and the relative humidity was 40% - 80%. The experiment started after 2 - 3 days of adaptive breeding.

[0029] 2. Instruments and Reagents

[0030] Electronic analytical balance (Shanghai Mettler - Toledo Co., Ltd.), Milli - Q ultrapure water machine (Millipore Corporation, USA), microplate reader (Bio - Tek Instruments, Inc., USA), refrigerated centrifuge (Thermo Fisher Scientific, USA), sterilized medical suture needles (Ningbo Medical Suture Co., Ltd.), tissue scissors, surgical forceps (Shanghai Surgical Instrument Factory), sodium pentobarbital (Sigma - Aldrich, USA). Compound 6 was provided by Suzhou Manyuan Biotechnology Co., Ltd., and dexamethasone (Dex) was purchased from Sigma - Aldrich, USA.

[0031] II. Experimental Methods

[0032] 1. Grouping and Administration of Experimental Animals

[0033] 120 male C57BL / 6J mice were randomly divided into 6 groups (6 mice per group):

[0034] (1) Sham - operation group: Intraperitoneal injection of an equal volume of normal saline 1 h before surgery, once.

[0035] (2) Model group: Intraperitoneal injection of an equal volume of normal saline 1 h before surgery, once.

[0036] (3) Low - dose compound 6 group: Intraperitoneal injection of compound 6 at a dose of 0.3 mg / kg 1 h before surgery, once.

[0037] (4) Medium - dose compound 6 group: Intraperitoneal injection of compound 6 at a dose of 1 mg / kg 1 h before surgery, once.

[0038] (5) High - dose compound 6 group: Intraperitoneal injection of compound 6 at a dose of 3 mg / kg 1 h before surgery, once.

[0039] (6) Positive drug group: Intraperitoneal injection of Dex at a dose of 2 mg / kg 1 h before surgery, once.

[0040] 2. Preparation of Cecal Ligation and Puncture (CLP) Model

[0041] C57BL / 6J mice were intraperitoneally anesthetized with sodium pentobarbital according to body weight. After verifying the absence of the righting reflex, the animals were placed in the supine position, and their four limbs were fixed on the operating board. The abdominal hair was shaved off. In the midline of the anterior abdomen, an incision about 1 cm was made along the linea alba, the epidermis and muscular layer were separated, the cecum was found in the abdominal cavity, the cecum and mesentery were separated, and 75% of the cecum was ligated with 5-0 suture. The cecum was penetrated once from the avascular side of the mesentery to the non-mesenteric side above the end of the cecum with a 20-gauge needle, and a small amount of intestinal contents was extruded from both ends of the needle hole. The cecum was then returned to the abdominal cavity, and the abdomen was sutured layer by layer to close the wound.

[0042] 3. Survival rate and body temperature detection

[0043] The survival rate of mice was continuously observed and calculated within 72 h, and the body temperature of the surviving mice was measured at 12 h, 24 h, 36 h, 48 h, 60 h, and 72 h, respectively.

[0044] 4. Data processing

[0045] All data were statistically analyzed using Graphpad Prism 8 software, expressed as mean±SD. The comparison between two groups was performed using Students’t test, and one-way ANOVA was used when comparing three or more groups. The Dunnett’s test was used for the test, and P<0.05 was considered statistically significant.

[0046] III. Experimental results

[0047] As Figure 1 shown, compared with the model group, different doses of Compound 6 could improve the survival rate of CLP mice and alleviate the decrease in body temperature. At 0.3 mg / kg and 1 mg / kg, the effect was slightly worse than that of the positive drug group, and at 3 mg / kg, the effect was better than that of the positive drug. In summary, the treatment of Compound 6 for sepsis showed a dose-dependence, and the best therapeutic effect was achieved at 3 mg / kg.

[0048] Example 2

[0049] Effects of different doses of Compound 6 on pathological damage of liver, kidney, lung, and intestinal tissues in sepsis mice induced by CLP surgery I. Experimental materials

[0050] The instruments and experimental animals were the same as those in Example 1.

[0051] II. Experimental methods

[0052] 1. Animal grouping, model establishment, and drug administration were the same as those in Example 1.

[0053] 2. HE staining of liver, kidney, lung, and intestinal tissues

[0054] After blood collection from each group of mice, the lung, intestine, liver, and kidney tissues were removed, fixed in 4% paraformaldehyde solution for 24 h, embedded in paraffin, and sent to Wuhan Sevier Biotechnology Co., Ltd. for HE staining. The sections were scanned using a digital scanner, and the HE sections were blindly scored by professionals.

[0055] 3. Data processing was the same as in Example 1.

[0056] III. Experimental Results

[0057] As Figure 2 shown in the results of B, 6-compound at different doses could improve the pathological damage of liver, kidney, lung, and intestinal tissues in septic mice induced by CLP surgery. At 0.3 mg / kg, 1 mg / kg, and 3 mg / kg, there were significant differences compared with the model group, and the effect was most obvious at 3 mg / kg. In summary, the treatment of 6-compound for sepsis was dose-dependent, and the treatment effect was optimal at 3 mg / kg. As Figure 2 shown in A, the H&E results also indicated that 6-compound (3 mg / kg) could reduce the pathological damage of liver, kidney, lung, and intestine.

[0058] Example 3

[0059] Effect of 6-compound at different doses on the expression of inflammatory factors in the serum of septic mice induced by CLP surgery

[0060] I. Experimental Materials

[0061] The instruments and experimental animals were the same as in Example 1. The interleukin-1β (IL-1β), chemokine (CXCL1), and interferon-β (IFN-β) kits were purchased from Nanjing Jinyibai Biotechnology Co., Ltd.

[0062] II. Experimental Methods

[0063] 1. Animal grouping, modeling, and administration were the same as in Example 1.

[0064] 2. Determination of the content of inflammatory factors in the serum of mice

[0065] After 24 h of modeling in each group of mice, blood was collected from the orbital cavity. After standing at room temperature for 30 min, centrifugation was performed (3500 rpm, 10 min), and the supernatant was taken and stored in aliquots. The determination of the contents of IFN-β, IL-1β, and CXCL1 in the serum of mice was carried out according to the kit instructions.

[0066] 3. Data processing was the same as in Example 1.

[0067] III. Experimental Results

[0068] As Figure 3As shown, compared with the sham operation group, the contents of IFN-β, IL-1β, and CXCL1 in the serum of mice in the model group were significantly increased. Compared with the model group, after administration of Compound 6, the contents of IFN-β, IL-1β, and CXCL1 in the plasma of mice were significantly decreased in a dose-dependent manner, and the therapeutic effect was optimal at 3 mg / kg. The above results indicate that Compound 6 can effectively inhibit the release of sepsis-related inflammatory factors induced by CLP.

[0069] Example 4

[0070] Effect of Different Doses of Compound 6 on the Expression of Damage-Related Molecules in the Serum of Mice with Sepsis Induced by CLP Surgery

[0071] I. Experimental Materials

[0072] The instruments, reagents, and experimental animals were the same as in Example 1. Kits for C-reactive protein (CRP), creatine kinase (CK), blood urea nitrogen (BUN), and alanine aminotransferase (ALT) were purchased from Nanjing Jinyibai Biotechnology Co., Ltd.

[0073] II. Experimental Methods

[0074] 1. Animal grouping, modeling, and administration were the same as in Example 1.

[0075] 2. Determination of the Contents of Damage-Related Molecules in the Serum of Mice

[0076] After 24 h of modeling in each group of mice, blood was collected from the orbital cavity. After standing at room temperature for 30 min, centrifugation was performed (3500 rpm, 10 min), and the supernatant was taken and stored in aliquots. The determination of the contents of CRP, CK, ALT, and BUN in the serum of mice was carried out according to the kit instructions.

[0077] 3. Data processing was the same as in Example 1.

[0078] III. Experimental Results

[0079] As Figure 4 shown, compared with the sham operation group, the contents of CRP, CK, ALT, and BUN in the serum of mice in the model group were significantly increased. Compared with the model group, after administration of Compound 6, the contents of CRP, CK, ALT, and BUN in the plasma of mice were significantly decreased in a dose-dependent manner, and the therapeutic effect was optimal at 3 mg / kg, all being better than the positive drug group. The above results indicate that Compound 6 can effectively improve the release of damage-related inflammatory factors in mice with sepsis induced by CLP.

Claims

1. Use of a compound as shown in the following formula in the preparation of a drug for the prevention and treatment of sepsis, 。 2. The use according to claim 1, characterized in that: The drug alleviates sepsis-induced organ damage.

3. The use according to claim 1, characterized in that: The drug alleviates sepsis-induced organ dysfunction.

4. The use according to claim 1, characterized in that: The drug reduces the release of inflammatory factors and damaging factors.

5. Use of a pharmaceutical composition in the preparation of a drug for the prevention and treatment of sepsis, characterized in that: The pharmaceutical composition comprises an active ingredient, and the active ingredient is shown in the following formula: 。 6. The use according to claim 5, characterized in that: The pharmaceutical composition also includes pharmaceutically acceptable excipients.

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