Use of Gelsemium for treating sepsis

Abstract

The invention relates to use of koumine, a gelsemium alkaloid monomer, for treating sepsis.

Description

Technical Field

[0001] The present invention relates to a new use of a gelatin alkaloid monomer koumine or a pharmaceutically acceptable salt thereof, and in particular to the use of koumine or a pharmaceutically acceptable salt thereof in preparing a pharmaceutical composition for treating sepsis. Background Art

[0002] Sepsis refers to the systemic inflammatory response syndrome (SIRS) caused by infection. In recent years, the incidence of sepsis has continued to rise worldwide, with an annual increase of 9%. The development of sepsis does not depend on the infected pathogenic microorganisms. Once it occurs, its occurrence and development follows its own pathological process and laws. Therefore, sepsis is essentially the body's response to infectious factors. The late inflammatory storm can lead to serious illness, making the clinical treatment of sepsis very difficult. Although anti-infection treatment and organ function support technology have made great progress in recent years, the mortality rate of sepsis is as high as 30% to 70%. There are also reports of mortality rates as high as 28%-50% (Xiao Wei, Yang Mingshi, Medical Review, Vol. 20, No. 8, pp. 1425-1429, April 2014).

[0003] Sepsis can be divided into sepsis, severe sepsis and septic shock according to its severity. Severe sepsis refers to sepsis accompanied by organ dysfunction, poor tissue perfusion or hypotension. Septic shock refers to severe sepsis accompanied by persistent hypotension that cannot be corrected after adequate fluid resuscitation, and is also considered a special type of severe sepsis.

[0004] The fundamental pathogenesis of sepsis is still unclear. It involves complex systemic inflammatory network effects, gene polymorphism, immune dysfunction, abnormal coagulation function, tissue damage, and abnormal host responses to different infectious pathogens and their toxins. It is closely related to the pathophysiological changes of multiple systems and organs in the body. Studies have shown that bacterial endotoxins can induce sepsis. The uncontrolled inflammatory response, immune dysfunction, high metabolic state, and multi-organ dysfunction in the pathophysiological process of sepsis can be directly or indirectly triggered by endotoxins. Infectious factors in sepsis activate the body's mononuclear macrophage system and other inflammatory response cells to produce and release a large number of inflammatory mediators. In sepsis, endogenous inflammatory mediators interact with each other to form a network effect and cause extensive damage to various systems and organs throughout the body. At the same time, certain cytokines, such as tumor necrosis factor-α (TNF-α), may play an important role in the occurrence and development of sepsis.

[0005] The complications of sepsis are essentially the clinical manifestations of the various stages of the pathophysiology of sepsis. Common complications include shock, acute lung injury / acute respiratory distress syndrome, etc.

[0006] At present, the clinical treatment of sepsis and related diseases is mainly through infection control or non-specific treatment such as glucocorticoids and symptomatic treatment such as prevention and treatment of organ failure and shock. Although it can relieve sepsis symptoms and prolong life in some patients, the effect is limited. Therefore, it is urgent to find new therapeutic drugs.

[0007] Koumine is the alkaloid monomer with the highest content in Chinese Gelsemium elegans Benth. Chinese patent CN 101323618 B discloses a method for extracting and isolating koumine from Gelsemium elegans plants. The inventors of this application discovered a new use of koumine in the treatment of sepsis during their research on the pharmacodynamics of koumine. Summary of the invention

[0008] In one aspect of the present invention, there is provided a use of koumine or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for treating sepsis in an individual in need thereof.

[0009] In another aspect of the present invention, there is provided koumine or a pharmaceutically acceptable salt thereof for use in treating sepsis in a subject in need thereof.

[0010] In another aspect of the present invention, a method for treating sepsis in an individual in need thereof is provided, the method comprising administering to the individual a therapeutically effective amount of koumine or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of a pharmaceutical composition comprising koumine or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 To show the effect of koumine on the survival rate of mice with cecal ligation and puncture (CLP)-induced sepsis.

[0012] Figure 2 The effect of koumine on the survival rate of mice with LPS-induced sepsis was shown.

[0013] Figure 3 The results showed that koumine could improve the pathological manifestations of liver, lung and kidney tissues in septic mice.

[0014] Figure 4 The results showed that koumine has a regulatory effect on the levels of inflammatory factors in the brain tissue of septic mice.

[0015] Figure 5 The results showed that koumine has a regulatory effect on the expression levels of NF-κB pathway-related proteins in septic mice. DETAILED DESCRIPTION

[0016] The present invention is now described in detail with reference to representative embodiments of the present invention. These embodiments are merely illustrative and should not be construed as limiting the scope of the present invention in any way. On the contrary, the present invention is intended to encompass all substitutions, modifications and equivalents within the scope of the present invention as defined by the claims.

[0017] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0018] The term "Hermaphrodite" used herein has the following structural formula, the molecular formula is C 20 H 22 N 2 O, which is an alkaloid monomer obtained from Gelsemium elegans, can also be obtained by synthesis:

[0019]

[0020] As used herein, the pharmaceutically acceptable salt of Gelsemium includes the salt formed by Gelsemium and organic acid or inorganic acid. Exemplary pharmaceutically acceptable salts include, but are not limited to: hydrochloride, sulfate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, acid phosphate, citrate, acetate, oxalate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate. Preferably, the pharmaceutically acceptable salt is Gelsemium hydrochloride.

[0021] As used herein, the term "pharmaceutically acceptable" means that the compound, composition or carrier is suitable for administration to a subject to achieve the uses or methods described herein without undue adverse side effects.

[0022] The term "subject" as used herein refers to a mammal or a human, including but not limited to primates (monkeys, chimpanzees, gorillas, etc.), rodents (such as rats, mice, gerbils, hamsters, ferrets and similar rodents), lagomorphs, suids (such as pigs, minipigs), equines, canines, felines, etc. In a preferred embodiment, the subject is a mammal. In a further preferred embodiment, the subject is a human.

[0023] The term "treatment" as used herein refers to the complete or partial cure or elimination of a disease, disorder or pathological state, including but not limited to one selected from the following, or a combination of two or more: alleviating or eliminating the cause of the disease, disorder or pathological state; improving or eliminating its pathological changes; alleviating or eliminating one or more symptoms and / or signs thereof; delaying or arresting its progression; reducing its severity; reducing its incidence; reducing its recurrence; and improving its prognosis.

[0024] The inventors of the present application have discovered through experiments that koumine or a pharmaceutically acceptable salt thereof (e.g., hydrochloride) has significant therapeutic efficacy on animal individuals (e.g., mice) suffering from sepsis, can improve the survival rate of the animal individuals, improve pathological changes / damages in one or more organs (including but not limited to bronchi, lungs, liver, and kidneys), reduce the levels of inflammatory factors in the animal individuals (especially brain tissue and blood), and inhibit the activation of the NF-κB signaling pathway.

[0025] Therefore, in one aspect, the present application provides a use of koumine or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for treating sepsis in an individual in need thereof.

[0026] In some embodiments, the administration of koumine or a pharmaceutically acceptable salt thereof can prevent, alleviate or eliminate one or more symptoms and / or signs of sepsis.

[0027] In some embodiments, the signs and symptoms include, but are not limited to:

[0028] 1) Typical symptoms and signs of sepsis, including:

[0029] Chills, fever (body temperature > 38.3°C) or hypothermia (body temperature < 36°C), palpitations (heart rate > 90 beats / min), shortness of breath (respiration > 20 times / min or arterial carbon dioxide partial pressure (PaCO 2 )<32mmHg), changes in mental status, obvious edema or positive fluid balance (more than 20ml / kg in 24 hours);

[0030] 2) Symptoms and signs of severe sepsis, including:

[0031] Hypotension, lactic acidosis, oliguria, acute changes in mental status, etc. related to one or more organ dysfunction and tissue hypoperfusion;

[0032] 3) Symptoms and signs of septic shock, including:

[0033] Hypotension that cannot be reversed by fluid therapy, impaired consciousness (such as anxiety, slow reaction or confusion), rapid pulse, cold and wet limbs, oliguria or anuria, etc.

[0034] In some embodiments, the administration of the koumine or a pharmaceutically acceptable salt thereof can improve the survival rate of the individual. For example, the survival rate is improved by more than about 1 to about 6 times, preferably about 1.5 to 5 times, more preferably about 2 to 4 times, for example about 2.5 to 3 times.

[0035] In some embodiments, the administration of koumine or a pharmaceutically acceptable salt thereof can delay or arrest the progression of the sepsis; in other embodiments, reduce the severity of the sepsis; and / or in some embodiments, improve the prognosis of the sepsis.

[0036] In some embodiments, the administration of koumine or a pharmaceutically acceptable salt thereof can prevent, improve or eliminate pathological changes and / or dysfunctions of one or more organs in the individual.

[0037] In some further embodiments, the organ is selected from the group consisting of thymus, lung, trachea, bronchus, brain, heart, liver, spleen, bone marrow, kidney and intestine, more preferably lung, bronchus, brain, liver and kidney, particularly preferably lung, bronchus, liver and kidney.

[0038] In some further embodiments, the pathological changes are selected from (e.g., lung) interstitial inflammatory cell infiltration, bronchial and alveolar dilatation, and alveolar fusion. The inflammatory cells may be, for example, lymphocytes, macrophages, monocytes, granulocytes (eosinophils, basophils, particularly neutrophils).

[0039] In some embodiments, the administration of koumine or a pharmaceutically acceptable salt thereof can reduce the level of inflammatory factors in the individual, particularly in the brain tissue and blood.

[0040] In some further embodiments, the inflammatory factor is one or more selected from tumor necrosis factor, interleukin, interferon and NO, preferably one or more selected from TNF-α, IL-1β, IL-6, IL-8, IL-18, INF-γ and NO, more preferably one or more selected from TNF-α, IL-1β and IL-6.

[0041] In some further embodiments, the inflammatory factor is one or more of TNF-α, IL-6 and IL-1β in blood and / or brain tissue.

[0042] In some embodiments, the administration of koumine or a pharmaceutically acceptable salt thereof can inhibit the activation of IKK phosphorylation in the individual; in some embodiments, inhibit the activation of IκBα phosphorylation and / or the degradation of IκBα in the individual; in some embodiments, inhibit the phosphorylation of p65 in the individual; and / or in some embodiments, inhibit the activation of the NF-κB signaling pathway in the individual.

[0043] In another aspect, the present application also provides use of koumine or a pharmaceutically acceptable salt thereof in preparing a pharmaceutical composition for preventing or treating complications of sepsis.

[0044] In some embodiments, the complication is one or more selected from shock, acute lung injury, acute respiratory distress syndrome, deep vein thrombosis, stress ulcers, metabolic acidosis, disseminated intravascular coagulation (DIC), acute renal failure, acute respiratory failure, and multiple organ dysfunction / failure.

[0045] In another aspect, the present application provides koumine or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising koumine or a pharmaceutically acceptable salt thereof, for use in: treating sepsis in an individual in need thereof; preventing, alleviating or eliminating one or more symptoms and / or signs of sepsis; improving the survival rate of the individual; delaying or arresting the progression of sepsis; reducing the severity of sepsis; improving the prognosis of sepsis; preventing, improving or eliminating pathological changes and / or dysfunction of one or more organs in an individual with sepsis; reducing the level of inflammatory factors in the individual; inhibiting the activation of IKK phosphorylation in the individual; inhibiting the activation of IκBα phosphorylation and / or the degradation of IκBα in the individual; inhibiting the phosphorylation of p65 in the individual; inhibiting the activation of NF-κB signaling pathway in the individual; and / or preventing or treating complications of sepsis.

[0046] In another aspect, the present application provides a method for treating sepsis in an individual in need thereof; preventing, alleviating or eliminating one or more symptoms and / or signs of sepsis; improving the survival rate of the individual; delaying or arresting the progression of sepsis; reducing the severity of sepsis; improving the prognosis of sepsis; preventing, improving or eliminating pathological changes and / or dysfunction of one or more organs in an individual with sepsis; reducing the level of inflammatory factors in the individual; inhibiting the activation of IKK phosphorylation in the individual; inhibiting the activation of IκBα phosphorylation and / or the degradation of IκBα in the individual; inhibiting the phosphorylation of p65 in the individual; inhibiting the activation of NF-κB signaling pathway in the individual; and / or preventing or treating complications of sepsis, the method comprising administering to the individual a therapeutically effective amount of koumine or a pharmaceutically acceptable salt thereof having the following structure, or a therapeutically effective amount of a pharmaceutical composition comprising koumine or a pharmaceutically acceptable salt thereof.

[0047] In various aspects described above, the sepsis includes severe sepsis and septic shock.

[0048] In some embodiments according to each aspect described above, the pharmaceutical composition comprises the kumquat or a pharmaceutically acceptable salt thereof as the sole active ingredient.

[0049] In other embodiments, the pharmaceutical composition further comprises one or more other active ingredients. In such embodiments, the koumine or its pharmaceutically acceptable salt and the one or more other active ingredients can be separately formulated so as to be respectively contained in a plurality of separate pharmaceutical compositions, or formulated together into a single pharmaceutical composition comprising them.

[0050] In some embodiments of the method of the present invention, the koumine or its pharmaceutically acceptable salt is administered to the individual as the sole active ingredient, or in combination with one or more other active ingredients. In the case of combined administration, the koumine or its pharmaceutically acceptable salt and the one or more other active ingredients can be administered to the individual simultaneously or sequentially. Combined administration includes the use of separate pharmaceutical compositions containing the koumine or its pharmaceutically acceptable salt and the one or more other active ingredients, respectively, administered simultaneously or sequentially in any order, or administering a single pharmaceutical composition containing the koumine or its pharmaceutically acceptable salt and the one or more other active ingredients.

[0051] The one or more other active ingredients are one or more chemotherapeutic agents, biological therapeutic agents, or a combination thereof known in the art to have therapeutic efficacy for sepsis or complications of sepsis.

[0052] The pharmaceutical composition optionally comprises one or more pharmaceutically acceptable carriers, such as excipients, disintegrants, diluents, binders, glidants, lubricants, pH regulators, preservatives, dispersants, suspending agents, ointment bases, emulsifiers, emollients, penetration enhancers, surfactants, propellants, flavoring agents, sweeteners, drug release regulators, etc. Those skilled in the art can select a suitable carrier according to the desired formulation form, administration method and release characteristics of the pharmaceutical composition.

[0053] The terms "comprising," "including," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude other unrecited elements or method steps, even though the other unrecited elements or method steps are not necessarily present (i.e., these terms also encompass the terms "consisting essentially of" and "consisting of.").

[0054] The pharmaceutical composition can be formulated into a solid, semisolid or liquid dosage form suitable for administration by any recognized administration method available in the art, including but not limited to: (1) suitable for oral administration: such as tablets, capsules, powders, granules, lozenges, aqueous or non-aqueous solutions or suspensions, syrups, etc.; (2) suitable for parenteral administration: such as subcutaneous, intramuscular or intravenous injections, such as sterile solutions or suspensions; (3) suitable for topical administration: such as plasters, ointments, creams, sprays or gels for skin or mucous membranes; (4) suitable for transdermal administration: such as patches, gel ointments, etc.; (5) suitable for vaginal or rectal administration: such as suppositories, emulsions, gels or effervescent tablets, etc.

[0055] As used herein, the term "therapeutically effective amount" refers to the amount of active ingredient that, when administered, will achieve the desired therapeutic effect for the stated use or method.

[0056] In purposes or methods as described herein, the dosage of the koumi element or its pharmaceutically acceptable salt and / or the one or more other active ingredients generally depends on multiple factors, including the severity of the individual, disease or condition treated, the speed of administration and the judgment of the prescribing physician. Generally speaking, effective dose is about 0.0001 to about 50mg per kg body weight per day, for example, about 0.0005 to about 25mg / kg / day or about 0.001 to about 5mg / kg / day (single or divided administration). For 70kg people, this will add up to about 0.007mg / day to about 3500mg / day, for example, about 0.035mg / day to about 1750mg / day or about 0.07mg / day to about 350mg / day. In some cases, the dosage level not higher than the lower limit of the aforementioned range can be enough, and in other cases, still can adopt larger doses without causing any harmful side effects, if necessary, the larger doses can be divided into several smaller doses to be administered throughout the day.

[0057] The content of koumine or its pharmaceutically acceptable salt and / or the one or more other active ingredients in the pharmaceutical composition or the amount used in the method can be about 0.001 mg to about 500 mg, such as about 0.01-200 mg, about 0.1-20 mg or about 0.5-10 mg, preferably about 0.1-20 mg.

[0058] The present invention is described in more detail below by way of examples, but these examples do not constitute any limitation to the present invention, and the scope of the present invention is only defined by the claims.

[0059] Example

[0060] Example 1: Effect of Gelsemium elegans on the survival rate of mice with sepsis induced by CLP

[0061] Healthy male 6-8 week old C57BL / 6 mice (weight 20-23 g), SPF grade, were purchased from Shanghai Slake Animal Experiment Co., Ltd., license: SCXK (Shanghai) 2017-0005. Adaptive feeding was performed for 5-7 days before the experiment, with free access to food and water, and maintained at a constant temperature (20-26°C) and constant humidity (40-70%) under a 12 / 12h light-dark cycle (lighting time 8:00-20:00). The experimental operation complied with the regulations issued by the International and Local Laboratory Animal Use and Protection Committee (hereinafter the same). 20 H 22 N 2 O·HCl, molecular weight 342.1499) powder was prepared by the School of Pharmacy of Fujian Medical University, batch number: KM201610 (purity 99.99% determined by HPLC), stored at 4°C away from light.

[0062] The animals were randomly divided into 5 groups, including a blank group, a model group, and an experimental group (a low-dose group, a medium-dose group, and a high-dose group), with 10 animals in each group. The experimental group was given different doses of koumine (0.8, 2.4, and 7.2 mg / kg) by intraperitoneal injection, and the blank group and the model group were given the same dose of normal saline. After the animals were anesthetized by intraperitoneal injection of 10% ketamine, the abdomen was wiped and disinfected with alcohol, and a 2 cm incision was made along the midline of the lower abdomen. The muscular layer, fascia, and peritoneal layer were opened layer by layer to expose the abdominal organs. The cecum was carefully removed with forceps, and a No. 4 intestinal thread was used to ligate the 1 / 2 of the end of the cecum. A No. 22 needle was used to perforate the hole, and a small part of the contents was gently squeezed out with forceps to ensure the patency of the hole, and the intestinal contents were pushed to the end of the cecum. The cecum was returned to the abdominal cavity, and the incision was closed and sutured. In the blank group, the cecum was only taken out and then put back into the abdominal cavity without ligation or perforation. The survival rate of the mice was observed and recorded every 12 hours. Taking the survival rate as an indicator, the effect of koumine on the survival rate of septic mice was observed.

[0063] The mice gradually woke up 4 hours after the cecal ligation surgery, and then it was found that the mice had reduced eating and activity, and had symptoms such as limb weakness and slow reaction. Figure 1 As shown, the survival rate of mice in the model group was as low as 12.5% ​​during the observation period (within 96 hours); the survival rate of mice in each koumine treatment group was higher than that in the model group, and there was a significant difference in the survival rate of mice between the experimental groups with different doses (***p < 0.01), among which the high-dose group (7.2 mg / kg) had the highest survival rate, reaching 72%. The results showed that koumine had a regulatory effect on sepsis in mice, and it was dose-dependent.

[0064] Example 2: Effect of Gelsemium elegans on the survival rate of mice with LPS-induced sepsis

[0065] Healthy male C57BL / 6 mice aged 6-8 weeks (weight 20-23 g), SPF grade, were purchased from Shanghai Slake Animal Experiment Co., Ltd., license: SCXK (Shanghai) 2017-0005. They were adaptively fed for 5-7 days before the experiment, with free access to food and water, and were raised at a constant temperature (20-26°C) and constant humidity (40-70%) under a 12 / 12h light-dark cycle (lighting time 8:00-20:00). 20 H 22 N 2 O·HCl, molecular weight 342.1499) powder was prepared by the School of Pharmacy of Fujian Medical University, batch number: KM201610 (purity 99.99% determined by HPLC), stored at 4°C away from light.

[0066] The animals were randomly divided into 5 groups, including a blank group, a model group, and an experimental group (low-dose group, medium-dose group, and high-dose group), with 10 animals in each group. The experimental group was given different doses of koumine (0.8, 2.4, and 7.2 mg / kg) by intraperitoneal injection, and the blank group was given the same dose of normal saline. 30 minutes later, LPS (15 mg / kg) was injected intraperitoneally into the model group and the experimental group. The survival rate of the mice was observed within 7 days, and the data were recorded every 12 hours. The effect of koumine on the survival rate of septic mice was observed using the survival rate as an indicator.

[0067] like Figure 2 As shown, mice in the model group and experimental group died one after another after being given LPS, and all mice in the model group (LPS group) died within 48 hours. The survival rate of mice given different doses of koumine was significantly improved (***p < 0.01), and the survival rate of mice in the high-dose group reached 77.8%. The results show that koumine has a therapeutic effect on sepsis in mice.

[0068] Example 3: Effect of Gelsemium elegans on the pathological manifestations of liver, lung and kidney tissues in septic mice

[0069] A CLP-induced sepsis mouse model was established according to the experimental protocol described in Example 1. 24 h after CLP surgery, mice were anesthetized by intraperitoneal incision of 4 cm along the midline of the abdomen, and part of the right lobe of the liver, kidney and right upper lobe of the lung were cut off and quickly placed in 10% formalin solution, followed by conventional ethanol dehydration, paraffin embedding, sectioning, and hematoxylin-eosin (HE) staining to observe the changes in the pathological morphology of liver, kidney and lung tissues.

[0070] like Figure 3As shown in the figure, HE staining showed that there were no inflammatory damage changes in the liver, lung and kidney tissues of the blank group mice; under the microscope, a large number of inflammatory cells infiltrated the lung interstitium of the model group mice, lymphocyte clusters were distributed around the bronchi, diffuse bronchi and alveolar expansion, and more alveolar fusion occurred. However, under the microscope, the lymphocyte clusters in the lung tissue of the mice in the koumine treatment group were reduced, and even partially disappeared. The expansion of the bronchi and alveoli was significantly reduced compared with the model group, and the alveolar fusion was also reduced. Compared with the blank group, the liver, kidney and lung of the model group mice were significantly damaged, while the symptoms of injury in the treatment group were alleviated compared with the model group, indicating that koumine has a therapeutic effect on liver, kidney and lung damage in septic mice.

[0071] Example 4: Regulatory effect of koumine on brain tissue and serum inflammatory factors in CLP-induced septic mice

[0072] According to the experimental scheme described in Example 1, a CLP-induced sepsis mouse model was established. 24 hours after the last administration, blood samples were collected by removing the eyeballs, standing at 4°C for 30 minutes, and then centrifuged at 3000r / min for 15 minutes, and stored after the serum was precipitated. At the same time, brain tissue was taken, and the levels of TNF-α, IL-1β and IL-6 in brain tissue and serum were determined by ELISA. The results are shown in Table 1 below.

[0073] like Figure 4 As shown in the figure, the levels of TNF-α and IL-1β in the brain tissue of CLP-induced septic mice (model group) increased significantly (###p<0.001), indicating that a more serious inflammatory response occurred in the brain of septic mice. Compared with the model group, different doses of koumine significantly reduced the levels of TNF-α and IL-1β in the brain tissue of CLP-induced septic mice (**p<0.01). The results showed that koumine had an inhibitory effect on brain tissue inflammation in CLP-induced septic mice.

[0074] The levels of TNF-α, IL-1β, and IL-6 in the serum of mice with CLP-induced sepsis model are summarized in Table 1 below.

[0075] Table 1: Levels of TNF-α, IL-1β and IL-6 in the serum of mice with CLP-induced sepsis model

[0076]

[0077] As shown in Table 1, compared with the blank control group, the levels of TNF-α, IL-1β and IL-6 in serum of the model group increased significantly, and the difference was extremely significant ( ###p<0.001), and the serum levels of TNF-α, IL-1β and IL-6 in the different doses of koumine groups were significantly reduced compared with the model group (**p<0.05, ***p<0.001). The results showed that koumine had a reducing effect on the levels of serum inflammatory factors in CLP-induced septic mice.

[0078] Example 5: Regulatory effect of koumine on the expression level of NF-κB pathway-related proteins in CLP-induced septic mice

[0079] According to the experimental scheme described in Example 1, a CLP-induced sepsis mouse model was established. 24 hours after CLP surgery, the mice were immediately decapitated, brain tissue was obtained, protein was extracted, and Western Blot was used to detect the expression levels of NF-κB pathway-related proteins. The results are shown in Figure 5 .like Figure 5 As shown in Figure 2, Western Blot results showed that the expression of p65 in the model group did not change significantly (p>0.05), but the degradation of NF-κB inhibitory protein IκBα increased significantly (p<0.01) ( Figure 5 ), and the phosphorylation of IKK, p65 and IκBα was also enhanced (**p<0.01)( Figure 5 ). Kosmine can inhibit the phosphorylation activation and degradation of IκBα by inhibiting the phosphorylation activation of IKK, inhibit the phosphorylation of downstream p65, and then inhibit the activation of NF-κB signaling pathway. Therefore, the inhibitory effect of Kosmine on the activation of NF-κB signaling pathway may be related to its treatment of sepsis.

[0080] In summary, koumine can increase the survival rate of mice with sepsis induced by LPS and CLP, significantly improve the pathological manifestations of liver, lung and kidney tissues of mice with sepsis, and significantly reduce the levels of inflammatory factors in brain tissue and serum of mice with sepsis. Secondly, koumine can inhibit the phosphorylation activation of IKK during sepsis, inhibit the phosphorylation activation and degradation of IκBα, inhibit the phosphorylation of downstream p65, and thus inhibit the activation of the NF-κB signaling pathway.

[0081] All publications and patent applications cited in this application are indicative of the state of the art in the field to which the present disclosure pertains. They are all incorporated herein by reference in their entirety.

[0082] According to the above description, those skilled in the art can determine the essential features of the present invention, and can make various changes and modifications to the present invention to make it suitable for various purposes and conditions without departing from the spirit and scope of the present invention. All such modifications and amendments are also included in the scope of the present invention. Therefore, the scope of the present invention should be determined by the appended claims and their legal equivalents.

Claims

1. Use of a koumine having the following structure or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for treating sepsis in an individual in need thereof: 。 2. The use according to claim 1, wherein the pharmaceutical composition is used for administration to treat one or more symptoms and / or signs of sepsis; improve the survival rate of the individual; delay or arrest the progression of sepsis; reduce the severity of sepsis.

3. The use according to claim 1 or 2, wherein the pharmaceutical composition is for administration to treat pathological changes and / or dysfunctions of one or more organs in the individual.

4. The use according to claim 3, wherein the organ is selected from the group consisting of lung, brain, liver, kidney.

5. The use according to claim 3, wherein the pathological changes are selected from interstitial inflammatory cell infiltration, bronchial and alveolar dilatation, and alveolar fusion.

6. The use according to any one of claims 1 or 2, wherein the pharmaceutical composition is used for administration to reduce the levels of inflammatory factors in brain tissue and blood.

7. The use according to claim 6, wherein the inflammatory factor is one or more of TNF-α, IL-1β and IL-6.

8. The use according to claim 7, wherein the inflammatory factor is one or more of TNF-α, IL-6 and IL-1β in blood and / or brain tissue.

9. The use according to any one of claims 1 or 2, wherein the pharmaceutical composition is for administration to inhibit activation of IKK phosphorylation in the individual; inhibit activation of IκBα phosphorylation and / or degradation of IκBα in the individual; inhibit phosphorylation of p65 in the individual; and / or inhibit activation of NF-κB signaling pathway in the individual.

10. Use of a koumine or a pharmaceutically acceptable salt thereof having the following structure in the preparation of a pharmaceutical composition for treating sepsis-related liver injury, sepsis-related lung injury, sepsis-related kidney injury and sepsis-related brain injury: 。 11. The use according to any one of claims 1, 2 or 10, wherein the sepsis includes severe sepsis and septic shock.

12. The use according to any one of claims 1, 2 or 10, wherein the only active ingredient in the pharmaceutical composition is the kumquat or a pharmaceutically acceptable salt thereof.

13. The use according to any one of claims 1, 2 or 10, wherein the pharmaceutical composition further comprises one or more other active ingredients.

14. The use according to any one of claims 1, 2 or 10, wherein the pharmaceutical composition optionally comprises one or more pharmaceutically acceptable carriers.

15. The method of claim 1 , wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, acid phosphate, citrate, acetate, oxalate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate.

16. The use according to any one of claims 1, 2 or 10, wherein the pharmaceutical composition is for oral administration, parenteral administration, topical administration, transdermal administration or rectal administration.

17. The use according to any one of claims 1, 2 or 10, wherein the pharmaceutical composition is in the form of a tablet, capsule, granule, syrup, patch, suppository, emulsion or gel.

18. The use according to any one of claims 1, 2 or 10, wherein the individual is a mammal or a human.

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