Application of CCL7 antagonist in preparation of medicine for treating sepsis

Abstract

The invention relates to application of a CCL7 antagonist in preparation of drugs for treating sepsis, it is found that CCL7 is highly expressed in serum of a sepsis patient in a high inflammatory reaction period, after the CCL7 antagonist is applied to a sepsis mouse in the high inflammatory reaction period, the death rate is remarkably reduced, the CCL7 antagonist is a CCL7 neutralizing antibody, and experiments prove that the CCL7 antagonist can be used for preparing drugs for treating sepsis. When the CCL7 antagonist is used for intravenous injection of mice according to an effective acting dose of 500 ug / kg / d, the death rate of mice in a CLP group after sepsis is 62%, and the death rate of mice in a CLP + CCL7nAb treatment group after sepsis is only 10%, so that the CCL7 antagonist is proved to be capable of effectively treating sepsis, reducing synthesis and release of in-vivo inflammatory factors and quickly promoting recovery of functions of visceral organs, and has important clinical treatment significance.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of a CCL7 antagonist in the preparation of a drug for treating sepsis. Background Art

[0002] Sepsis is a systemic inflammatory response, and severe cases can lead to multiple organ dysfunction and even death. Sepsis is a major public health problem of global concern, imposing a huge disease burden. According to the statistics of the World Health Organization, more than 3 million people die from sepsis globally every year, accounting for about 10% of the total death toll. Sepsis is also one of the important reasons for the consumption of health resources, and its treatment costs are high. In addition, sepsis has a serious impact on the quality of life, working ability and family economic status of patients.

[0003] Regarding the treatment of sepsis, it mainly includes aspects such as anti-infection, fluid resuscitation and multi-organ function support. Anti-infection is the core of sepsis treatment, and the early application of broad-spectrum antibiotics is a recommended strategy. However, due to the abuse and irrational use of antibacterial drugs, drug-resistant strains have emerged. In addition, fluid resuscitation is also an important part of sepsis treatment, aiming to maintain effective blood volume and perfused organs. However, the specific methods and goals of fluid resuscitation are still controversial and need to be individualized based on the patient's own situation. For the supportive treatment of multiple organ dysfunction, mechanical ventilation and renal replacement therapy are crucial. However, long-term mechanical ventilation can cause complications, such as barotrauma of the alveoli, continuous high airway pressure, especially high PEEP, which can affect the return blood volume, respiratory tract infection, laryngeal injury, neonatal pulmonary-bronchial dysplasia, etc. Patients undergoing renal replacement therapy need to take immunosuppressants for a long time after surgery, the treatment is expensive and the operation is complex. Although there are many current treatment methods for sepsis, each method has certain limitations, and the mortality and long-term prognosis of sepsis patients have not been significantly improved through intensive treatment. Therefore, finding new drugs that can effectively reduce the mortality of sepsis and have good safety is an urgent problem to be solved in the current treatment of sepsis.

[0004] Chemokine Ligand 7 (CCL7), also known as Monocyte Chemoattractant Protein 3 (MCP-3), is a chemokine. The gene encoding CCL7 is located on human chromosome 17q11.2-q12 and contains 3 exons and 2 introns. The expression of CCL7 is affected by the stimulation of various inflammatory mediators, such as tumor necrosis factor-α and interleukin-1β. In addition, the transcriptional level of CCL7 is also regulated by nuclear factor κB. Many types of cells can produce CCL7, including monocytes, macrophages, lymphocytes, and endothelial cells. CCL7 mainly attracts and activates monocytes. By binding to specific G protein-coupled receptors CCR1, CCR2, and CCR3 on the cell surface, it generates specific signal transduction, promotes the chemotaxis, migration, adhesion, and proliferation of monocytes to the inflammatory site, and then produces a large number of inflammatory mediators, leading to inflammatory damage in various organs of the body. Summary of the Invention

[0005] In view of the deficiencies in the prior art, the present invention provides the use of a CCL7 antagonist in the preparation of a drug for treating sepsis.

[0006] The technical solution of the present invention is as follows:

[0007] The use of a CCL7 antagonist in the preparation of a drug for treating sepsis.

[0008] Further, the CCL7 antagonist is a CCL7 neutralizing antibody.

[0009] Further, the drug can reduce the content of CCL7 in the serum of sepsis patients.

[0010] Further, the effective dose of the drug is 500 μg / kg / d.

[0011] Further, the administration route of the drug is intravenous injection.

[0012] Further, the drug also includes pharmaceutically acceptable excipients, and the pharmaceutically acceptable excipients include one or more of diluents, wetting agents, binders, disintegrants, lubricants, color, flavor and fragrance regulators, solvents, solubilizers, cosolvents, emulsifiers, antioxidants, metal chelating agents, inert gases, preservatives, local anesthetics, pH regulators, isotonic regulators, and isosmotic regulators.

[0013] Further, the dosage form of the drug is an injection.

[0014] The drug of the present invention can also be used in combination with other drugs for treating sepsis, and the combination of multiple drugs can greatly improve the success rate of treatment.

[0015] Compared with the prior art, the present invention has at least the following advantages:

[0016] The present invention relates to the application of a CCL7 antagonist in the preparation of a drug for treating sepsis. The present invention discovers that CCL7 is highly expressed in the serum of sepsis patients during the hyperinflammatory response phase. After applying the CCL7 antagonist to sepsis mice in the hyperinflammatory response phase, the mortality rate is significantly reduced. The CCL7 antagonist is a CCL7 neutralizing antibody. It has been experimentally verified that when intravenously injecting mice with an effective dose of 500 μg / kg / d, the 7-day mortality rate of mice in the CLP group after sepsis is 62%, while the 7-day mortality rate of mice in the CLP + CCL7 nAb treatment group after sepsis is only 10%. This proves that the CCL7 antagonist can effectively treat sepsis, reduce the synthesis and release of inflammatory factors in the body, and rapidly promote the recovery of the functions of various organs, which has important clinical therapeutic significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the specific embodiments of the present invention, the drawings required for use in the specific embodiments or the description of the prior art will be briefly introduced below.

[0018] Figure 1 It shows the difference in the expression level of CCL7 in the serum of sepsis patients during the hyperinflammatory response phase and the recovery phase in Example 1 of the present invention;

[0019] Figure 2 It shows the difference in the expression level of CCL7 in the serum of sepsis mice during the hyperinflammatory response phase and the recovery phase in Example 2 of the present invention;

[0020] Figure 3 It shows the change in the mortality rate of sepsis mice in the hyperinflammatory response phase after injecting the CCL7 neutralizing antibody in Example 3 of the present invention. SPECIFIC EMBODIMENTS

[0021] The present invention will be further described in detail below. It is necessary to point out here that the following specific embodiments are only used to further illustrate the present invention and cannot be understood as limiting the protection scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention based on the above application content.

[0022] The present invention generally and / or specifically describes the materials and test methods used in the experiments. The test methods or testing methods involved, unless otherwise specified, are conventional methods; the reagents or instruments used, unless the manufacturer is indicated, are conventional products commercially available and are prepared or used by conventional methods.

[0023] Source of materials:

[0024] CCL7 neutralizing antibody: Mouse CCL7 / MCP-3 / MARC Antibody, catalog number AF-456-NA, purchased from company R&D.

[0025] Example 1: CCL7 is highly expressed in the serum of patients in the hyperinflammatory phase of sepsis

[0026] 1. Research protocol

[0027] (1) Sample source: Trauma patient biobank and clinical research database of Army Characteristic Medical Center

[0028] (2) Experimental group: 40 serum samples from patients with sepsis in the hyperinflammatory phase

[0029] (3) Control group: 38 serum samples from patients with sepsis in the convalescent phase

[0030] (4) Detection method: Detect the expression level of CCL7 in the experimental group and the control group by liquid chip technology

[0031] (5) Statistical analysis: If the two groups of data meet the requirements of normality and homogeneity of variance, Student's t-test is selected; when the two groups only meet normality but not homogeneity of variance, Weltch's t-test is selected; when the two groups do not meet normality, Wilcoxon rank sum test or Wilcoxon-Mann-Whitney test is selected.

[0032] 2. Research results

[0033] The expression levels of CCL7 in the serum of patients with sepsis in the hyperinflammatory phase and the convalescent phase are shown in Figure 1 , it can be seen from the figure that the expression level of CCL7 in the serum of patients with sepsis in the hyperinflammatory phase (mean ± standard deviation: 16.31 ± 4.59 pg / ml) is significantly higher than that in the convalescent phase (mean ± standard deviation: 13.5 ± 2.39 pg / ml), P = 0.0012, and the difference is statistically significant.

[0034] Example 2: CCL7 is highly expressed in the serum of mice in the hyperinflammatory phase of sepsis

[0035] 1. Research protocol

[0036] (1) Experimental animals: 7-week-old male C57bl / 6j mice (body weight 18 - 22 g)

[0037] (2) Experimental group: 8 serum samples from mice during the hyperinflammatory phase after CLP modeling (sepsis model);

[0038] (3) Control group: 11 serum samples from mice during the convalescent phase after CLP modeling (sepsis model);

[0039] (4) Detection method: Detect the expression level of CCL7 in the experimental group and the control group by liquid chip technology;

[0040] (5) Statistical analysis: If the two groups of data meet the requirements of normality and homogeneity of variance, Student's t test is selected; when the two groups only meet normality but not homogeneity of variance, Weltch's t test is selected; when the two groups do not meet normality, Wilcoxon rank sum test or Wilcoxon-Mann-Whitney test is selected.

[0041] 2. Research results

[0042] The expression levels of CCL7 in the sera of septic mice during the hyperinflammatory phase and convalescent phase are shown in Figure 2 , and it can be seen from the figure that the expression level of CCL7 in the sera of septic mice during the hyperinflammatory phase (mean ± standard deviation: 1622.4 ± 1335.3 pg / ml) was significantly higher than that during the convalescent phase (mean ± standard deviation: 672.7 ± 460.4 pg / ml), P = 0.04, and the difference was statistically significant.

[0043] Example 3 Neutralizing antibody against CCL7 significantly reduces the mortality of septic mice

[0044] 1. Research plan

[0045] (1) Experimental animals: Male C57bl / 6j mice, 7 weeks old, weighing 18 - 22 g;

[0046] (2) Experimental group (CLP + CCL7nAb group): After CLP modeling (sepsis model) in mice during the hyperinflammatory phase, inject neutralizing antibody against CCL7 into the tail vein of mice, 500 μg / kg, once;

[0047] (3) Control group:

[0048] ① Isotype control group (CLP + IgG group): After CLP modeling in mice during the hyperinflammatory phase, inject IgG into the tail vein of mice, 500 μg / kg, once; ② Negative control group (CLP group): After CLP modeling in mice during the hyperinflammatory phase, no drug treatment is given;

[0049] (4) Detection index: Mortality rate 7 days after sepsis;

[0050] (5) Statistical analysis: Survival analysis.

[0051] 2. Research results

[0052] The survival curves of the experimental group and the two control groups are as Figure 3 shown. The mortality rate of mice in the CLP+CCL7 nAb treatment group 7 days after sepsis was 10%. The mortality rate of mice in the isotype control group (CLP+IgG group) 7 days after sepsis was 60%. The mortality rate of mice in the negative control group (CLP group) 7 days after sepsis was 62%. The mortality rate of mice in the treatment group was significantly lower than that in the isotype control group and the negative control group, and the difference was statistically significant.

[0053] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered by the scope of the claims and the description of the present invention.

Claims

1. Use of a CCL7 antagonist in the preparation of a drug for treating sepsis.

2. The application according to claim 1, characterized in that, The CCL7 antagonist is a CCL7 neutralizing antibody.

3. The application according to claim 2, characterized in that, The drug can reduce the content of CCL7 in the serum of sepsis patients.

4. The application according to claim 3, characterized in that, The effective dose of the drug is 500 ug / kg / d.

5. The application according to any one of claims 1 to 3, characterized in that The administration route of the drug is intravenous injection.

6. The application according to claim 5, characterized in that, The drug also includes pharmaceutically acceptable excipients, and the pharmaceutically acceptable excipients include one or more of diluents, wetting agents, binders, disintegrants, lubricants, color, flavor and odor regulators, solvents, solubilizers, cosolvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local anesthetics, pH regulators, isotonic regulators and isosmotic regulators.

7. The application according to claim 6, wherein The dosage form of the drug is an injection.

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