Targeted heparanase inhibitor for treating sepsis and application of targeted heparanase inhibitor

Abstract

The invention belongs to the technical field of targeted inhibitors, and particularly provides a targeted heparanase inhibitor for treating sepsis and application of the targeted heparanase inhibitor, the targeted heparanase inhibitor comes from natural medicine components, is simple to synthesize and has better inhibition activity, and the problems that existing heparanase inhibitors are not multiple in variety, not comprehensive in application and poor in inhibition effect are solved. Or is difficult to synthesize or obtain.

Description

Technical Field

[0001] The present invention belongs to the technical field of targeted inhibitors, and particularly relates to an inhibitor targeting heparanase for treating sepsis and its application. Background Art

[0002] Sepsis is an infection caused by bacteria, fungi, viruses, parasites, etc., leading to an inflammatory response in the body. If the inflammatory response intensifies, it will cause a deep inhibition of immune function. The main symptoms of sepsis include chills, fever, palpitation, shortness of breath, mental state changes, etc. Sepsis can develop into severe sepsis and septic shock, leading to organ dysfunction and circulatory disorders. The latest global epidemiological survey shows that the fatality rate of sepsis is 26.7% - 41.9%, which greatly endangers the lives of patients.

[0003] Heparanase (HPA) is currently the only known enzyme that can cleave heparan sulfate side chains and has been widely studied in aspects such as tumor metabolism, procoagulant activity, angiogenesis, and inflammation. In recent years, heparanase has been proven to have a very important impact in sepsis, especially sepsis-related acute kidney injury (S-AKI). Therefore, developing inhibitors targeting heparanase has become a new direction for treating sepsis.

[0004] Patent 2007100994479 discloses a small molecule polypeptide inhibitor of human heparanase, and these small molecule polypeptide inhibitors have potential medical value in the targeted treatment of tumors. Patent 2017800086184X discloses a quinazoline compound and introduces the use of this type of compound in treating diseases related to heparanase activity. Currently, there is literature proving (Role of heparanase in sepsis-related acute kidney injury) that inhibitors of heparanase are related to sepsis-related acute kidney injury, and heparanase inhibitors have good therapeutic and alleviating effects on sepsis. Therefore, developing heparanase inhibitors has become a new idea for treating sepsis. However, more structures and types of heparanase inhibitors remain to be developed. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide an inhibitor targeting heparanase for treating sepsis and its application. It is derived from natural drug components, is simple to synthesize, and has better inhibitory activity, so as to solve the problems that the existing types of heparanase inhibitors are not enough, the applications are not comprehensive enough, or they are difficult to synthesize or obtain.

[0006] To achieve the above object, the technical solution adopted by the present invention is: a heparanase-targeted inhibitor for treating sepsis, the inhibitor can achieve the effect of treating sepsis by inhibiting heparanase, and the inhibitor is chlorogenic acid, and its chemical name is (1R,3R,4S,5R)-3-[(E)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy-1,4,5-trihydroxycyclohexane-1-carboxylic acid, and the molecular formula of this compound is:

[0007]

[0008] On the basis of the above technical solution, the present invention can also have the following further specific selections or optimized selections.

[0009] Specifically, the sepsis is liver severe sepsis, lung severe sepsis, and kidney severe sepsis.

[0010] The present invention also provides the use of chlorogenic acid in the preparation of drugs for treating or preventing sepsis.

[0011] Specifically, the chlorogenic acid effectively inhibits the expression of inflammatory factors by inhibiting the activity of heparanase.

[0012] Preferably, the chlorogenic acid improves the case injury of CLP-induced sepsis by inhibiting the activity of heparanase.

[0013] Specifically, the dosage form of the drug is oral administration.

[0014] Specifically, the dosage of the drug is 30-60 mg / kg.

[0015] The research premise of the present invention is that some studies have reported that heparanase plays an important role in promoting pathological processes such as tumor growth, metastasis, angiogenesis, thrombosis, fibrosis, inflammation, autoimmunity, and renal dysfunction. And it has been found in studies that the expression of heparanase is increased in tissues such as the kidneys, livers, and lungs of mice with sepsis. Therefore, inhibiting the activity of heparanase has become a new idea and new direction for preventing or treating sepsis, especially liver severe sepsis, lung severe sepsis, and kidney severe sepsis.

[0016] On this premise, this research team is committed to screening a variety of natural drug components and selecting inhibitors that can inhibit heparanase. Among many natural drug extracts (compounds such as chlorogenic acid, isochlorogenic acid, lonicerin, biflorol, linalool, etc.), chlorogenic acid has been screened out to have heparanase inhibitory activity. According to the research findings, this is the first time to discover the heparanase inhibitory activity of chlorogenic acid, and its IC 50The value can reach below 30 μM. Experiments have proven that chlorogenic acid has low toxic and side effects in organisms and can effectively improve the pathological damage of CLP-induced sepsis. The above content not only expands the new direction for the development of small molecule inhibitors of heparanase, but also represents a new starting point and new use for the application of natural drug products, with important scientific research value and good clinical application prospects. Brief Description of the Drawings

[0017] Figure 1 In vitro inhibitory activity of chlorogenic acid on heparanase. Detailed Implementation Modes

[0018] To better understand the present invention, the content of the present invention will be further clarified below in conjunction with the drawings and specific embodiments, but the content of the present invention is not limited to the following embodiments.

[0019] Example 1: Obtaining Chlorogenic Acid

[0020] The present invention provides an inhibitor targeting heparanase for the treatment of sepsis. The inhibitor can achieve the effect of treating sepsis by inhibiting heparanase. The inhibitor is chlorogenic acid, and its chemical name is (1R,3R,4S,5R)-3-[(E)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy-1,4,5-trihydroxycyclohexane-1-carboxylic acid. The molecular formula of this compound is:

[0021]

[0022] The above compound is obtained by catalytic hydrolysis of methyl (1S)-1-ethoxycarbonyloxy-3-cis-[3,4-dihydroxy-trans-cinnamoyloxy]-4′-trans-5′-trans-dihydroxycyclohexane-γ-carboxylate in methanol with barium hydroxide, or chlorogenic acid is isolated from green coffee beans. The chlorogenic acid used in the examples of the present invention is purchased from Changsha Shanghe Biotechnology Co., Ltd.

[0023] Example 2: Inhibition of Heparanase

[0024] According to the standard enzymatic assay disclosed in the literature (Development of a colorimetric assay for heparanase activity suitable for kinetic analysis and inhibitor screening), the IC 50 value of the heparanase inhibitory compound of the present invention can be tested. Among them, heparanase is purchased from Hubei Yuansheng Peptide Biotechnology Co., Ltd. For the enzymatic assay, the test compound is dissolved and added to the assay at different concentrations to calculate the inhibition level, as shown in the appendixFigure 1 As shown. According to experimental evidence, the half-maximal inhibitory concentration (IC 50 ) of chlorogenic acid on heparanase is approximately 10.7 μM.

[0025] It should be noted that since the screening experiment of the present invention is the screening of the inhibitory activities of multiple compounds on heparanase, the half-maximal inhibitory concentration is only for heparanase, and subsequent research experiments will also carry out tests on the activities of other enzymes.

[0026] Example 3: Inhibition of Chlorogenic Acid on Inflammatory Factors in Sepsis

[0027] Forty SPF-grade male C57BL / 6 mice, 8 - 10 weeks old, weighing 25 ± 2 g, were purchased from Cyagen Biosciences (Suzhou) Co., Ltd. Thirty of them were randomly selected and used to establish a model group according to the protocol proposed in "Establishment and Evaluation of a Mouse Sepsis Model" (Jing Xizhong, Jia Huanhuan, Luo Ting, et al. Establishment and Evaluation of a Mouse Sepsis Model [J]. Acta Laboratorium Animalis Scientia Sinica, 2016, 24(2): 158 - 163.); the remaining 10 were used as a blank control group. The model group was randomly divided into 3 groups, with 10 mice in each group. One group was not treated at all and served as a model control group; the other 2 groups (Model Group 1 and 2) were each orally administered 5 ml of the chlorogenic acid described in Example 1 of the present invention twice a day, once in the morning and once in the evening. The dosing amounts of Model Group 1 and 2 were calculated as 30 mg / kg and 60 mg / kg respectively. Carotid blood of each group of mice was collected after 48 h and 96 h, and the expression levels of tumor necrosis factor α (TNF-α), inflammatory factors IL-6, and IL-1β were detected, and the average values were calculated. The results are shown in Table 1.

[0028] Table 1 Effects of Different Treatments on Inflammatory Factors and Tumor Necrosis Factor

[0029]

[0030] As can be seen from the above table, compared with the blank control group, the expression levels of tumor necrosis factor α (TNF-α) and inflammatory factors IL-6 and IL-1β in the model control group were significantly increased, indicating that the mouse sepsis model was successfully established. After treatment with the chlorogenic acid described in Example 1 of the present invention, the expression levels of tumor necrosis factor α (TNF-α) and inflammatory factors IL-6 and IL-1β in Model Group 1 and 2 decreased, indicating that the chlorogenic acid described in Example 1 of the present invention can significantly reduce the body's inflammatory response. By comparing Model Group 1 and 2 after treatment with the chlorogenic acid described in Example 1 of the present invention, it can be seen that the treatment effect of the high-dose group of mice is relatively higher, and no adverse reactions have been found for the time being.

[0031] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. An inhibitor targeting heparanase for treating sepsis, characterized in that: The compound is called chlorogenic acid and its chemical structure is:

2. The heparanase-targeted inhibitor for treating sepsis according to claim 1 is characterized in that: The sepsis is severe liver sepsis.

3. The heparanase-targeted inhibitor for treating sepsis according to claim 1 is characterized in that: The sepsis is severe pulmonary sepsis.

4. The inhibitor targeting heparanase for treating sepsis according to claim 1 is characterized in that: The sepsis is severe renal sepsis.

5. Application of chlorogenic acid in the preparation of drugs for treating or preventing sepsis.

6. The use according to claim 5, characterized in that: The chlorogenic acid effectively inhibits the expression of inflammatory factors by inhibiting the activity of heparanase.

7. The use according to claim 5, characterized in that: The chlorogenic acid improves the case damage of CLP-induced sepsis by inhibiting the activity of heparanase.

8. The use according to claim 5, characterized in that: The dosage form of the drug is oral administration.

9. The use according to claim 5, characterized in that: The dosage of the drug is 30-60 mg / kg.

10. The use according to claim 5, characterized in that: The sepsis is severe liver sepsis, severe lung sepsis, or severe kidney sepsis.

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