Application of polycyclic polyisopentenyl phloroglucinol compound Atenuamione D in preparation of medicine for treating hepatocellular carcinoma

Abstract

The invention relates to an application of a polycyclic polyisopentenyl phloroglucinol compound Atenuamione D, which is extracted and separated from Hypericum sampsonii Hance, in preparation of a medicine for treating hepatocellular carcinoma, in particular to an application of the polycyclic polyisopentenyl phloroglucinol compound Atenuamione D in preparation of a medicine for treating hepatocellular carcinoma. The in-vivo pharmacological research shows that the polycyclic polyisopentenyl phloroglucinol compound can obviously reduce the tumor weight of nude mice with hepatocellular carcinoma under the dosage of 25 mg / kg and 50 mg / kg, can reverse the purine metabolism abnormality of nude mice with hepatocellular carcinoma, and can be used for preparing the medicine for treating hepatocellular carcinoma.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a polycyclic polyisoprenyl phloroglucinol compound extracted and separated from the aboveground part of Hypericum sampsonii Hance, a preparation method thereof and application of the polycyclic polyisoprenyl phloroglucinol compound in preparation of a drug for treating hepatocellular carcinoma. BACKGROUND

[0002] Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer, accounting for 75-90% of all liver cancer cases, which mainly originates from the malignant transformation of hepatocytes, and its clinical manifestations mainly include liver pain, liver function abnormalities, metastasis symptoms and cancer syndrome, etc. The incidence of HCC is closely related to chronic liver diseases (such as cirrhosis and viral hepatitis), and HCC has high invasiveness and metastasis tendency, and the early symptoms are occult, and the prognosis in the middle and late stages is poor.

[0003] At present, the main treatment for HCC is chemotherapy, but the complex tumor microenvironment, tumor heterogeneity and genetic mutations and other factors limit the effect of existing therapies. In recent years, multi-target kinase inhibitors, sorafenib, atezolizumab, bevacizumab and lenvatinib and other systemic chemotherapy drugs have been approved as first-line treatment options for advanced HCC. However, these therapies can improve patient survival, but can also cause serious side effects such as hypertension, diarrhea, loss of appetite, weight loss and other serious side effects, further damaging the overall health of patients. In addition, the current clinic is still facing the problem of drug resistance of many existing anticancer drugs. Therefore, accelerating the development of new drugs for treating hepatocellular carcinoma has become a major challenge that needs to be addressed in the clinic.

[0004] Natural small molecules are an important source of antitumor drugs, and according to statistics, 49% of existing anticancer small molecule drugs are related to natural small molecules or their derivatives. China has rich resources of medicinal plants, which have rich structural diversity and biocompatibility, and many drug source molecules have been found from natural small molecules. Therefore, finding natural small molecule drugs for treating HCC from medicinal plants to meet the clinical demand is an important way to develop new anti-HCC drugs. SUMMARY

[0005] Therefore, the purpose of the present application is to provide a polycyclic polyisoprenyl phloroglucinol compound and a preparation method thereof and application of the polycyclic polyisoprenyl phloroglucinol compound in preparation of a drug for treating hepatocellular carcinoma. Since the polycyclic polyisoprenyl phloroglucinol compound is derived from biological sources, when it is prepared into a drug for treating hepatocellular carcinoma, it has a significant anticancer effect and good biocompatibility with the human body, which is beneficial to reducing the side effects of the drug on the human body.

[0006] To solve the technical problems of the present application, the present application provides the following technical solutions:

[0007] Use of polycyclic polyisoprenyl depsidate attenuatumione D in preparation of a medicine for treating hepatocellular carcinoma, wherein the polycyclic polyisoprenyl depsidate attenuatumione D has the following chemical structure:

[0008]

[0009] Further, the polycyclic polyisoprenyl depsidate attenuatumione D has a content of 0.1-95wt% in the medicine.

[0010] Further, the polycyclic polyisoprenyl depsidate attenuatumione D is obtained by extraction and separation from the Herba Hyperici Japonici.

[0011] Further, the preparation method of the polycyclic polyisoprenyl depsidate attenuatumione D comprises the following steps:

[0012] S1: mixing the Herba Hyperici Japonici powder with 8 times of 50-100% ethanol, heating and refluxing for extraction for 1h, extracting for 3 times, filtering and combining the 3 times of extraction solutions, and reducing pressure to concentrate to a thick solution to obtain Herba Hyperici Japonici total extract;

[0013] S2: passing the Herba Hyperici Japonici total extract through a D101 macroporous resin column, using ethanol and water as eluent, gradient eluting from 80% ethanol to 100% ethanol, collecting the eluent of the 100% ethanol section, and rotary evaporating and concentrating to remove the solvent to obtain a mixture A after concentration;

[0014] S3: separating the mixture A by preparative HPLC of Rp-C18 reverse-phase silica gel filler, wherein the mobile phase is 80% acetonitrile, the flow rate is 8.0ml / min, the peak with a retention time of 18.6min is collected, and rotary evaporation and concentration are performed to remove the solvent to obtain the polycyclic polyisoprenyl depsidate attenuatumione D.

[0015] Further, the medicine comprises a pharmaceutically acceptable excipient.

[0016] Further, the medicine is any one of a liquid preparation, a solid preparation, a semi-solid preparation, and a gaseous preparation.

[0017] Further, the medicine is any one of an oral liquid, a tablet, a capsule, a dripping pill, a granule, or an injection.

[0018] Further, the effective administration dose of the medicine is 25-50mg / kg / day.

[0019] The compound of the present application or the pharmaceutical composition containing it can be administered in unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0020] The administration dosage form can be a liquid dosage form, a solid dosage form or a semi-solid dosage form. The liquid dosage form can be a solution (including true solution and colloidal solution), an emulsion (including o / w type, w / o type and multiple emulsion), a suspension, an injection (including water injection, powder injection and infusion), eye drops, nose drops, lotion and liniment, etc.; the solid dosage form can be a tablet (including ordinary tablet, enteric-coated tablet, chewable tablet, dispersible tablet, effervescent tablet, oral disintegrating tablet), a capsule (including hard capsule, soft capsule, enteric-coated capsule), a granule, a powder, a pellet, a drop pill, a suppository, a film, a patch, an aerosol (powder) mist, a spray, etc.; the semi-solid dosage form can be an ointment, a gel, a paste, etc.

[0021] The compound of the present application can be prepared into ordinary preparation, sustained-release preparation, controlled-release preparation, targeted preparation and various microparticle drug delivery systems.

[0022] In order to prepare the compound of the present application into a tablet, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, solubilizers. The diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agents can be water, ethanol, isopropyl alcohol, etc.; the binders can be starch paste, dextrin, sugar syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecylsulfate, etc.; the lubricants and solubilizers can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0023] The tablet can be further prepared into a coated tablet, such as sugar-coated tablet, film-coated tablet, enteric-coated tablet, or a double-layer tablet and a multi-layer tablet.

[0024] To make the administration unit into a capsule, the effective component compound of the present application can be mixed with a diluent, a co-solvent, and the mixture can be directly placed into a hard capsule or a soft capsule. The effective component compound of the present application can also be first mixed with a diluent, a binder, and a disintegrant to form a granule or a pellet, and then placed into a hard capsule or a soft capsule. The various diluents, binders, wetting agents, disintegrants, and co-solvents used for preparing tablets of the compound of the present application can also be used for preparing capsules of the compound of the present application.

[0025] To make the compound of the present application into an injection, water, ethanol, isopropyl alcohol, propylene glycol, or a mixture thereof can be used as a solvent, and an appropriate amount of a commonly used solubilizer, a co-solvent, a pH adjuster, and an osmotic pressure adjuster in the art can be added. The solubilizer or the co-solvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH adjuster can be a phosphate, an acetate, hydrochloric acid, sodium hydroxide, etc.; and the osmotic pressure adjuster can be sodium chloride, mannitol, glucose, a phosphate, an acetate, etc. If a freeze-dried powder injection is prepared, mannitol, glucose, etc. can also be added as a supporting agent.

[0026] In addition, if necessary, a coloring agent, a preservative, a flavoring agent, or other additives can also be added to the pharmaceutical preparation.

[0027] To achieve the purpose of medication and enhance the therapeutic effect, the drug or the pharmaceutical composition of the present application can be administered by any known administration method.

[0028] The administration dose of the pharmaceutical composition of the compound of the present application can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the administration route, and the dosage form, etc. Generally, the suitable dose of the compound of the present application per day is in the range of 0.001-50 mg / Kg body weight. The above dose can be administered in one dosage unit or divided into several dosage units, which depends on the clinical experience of the doctor and the administration scheme including the use of other therapeutic methods.

[0029] The compound or the composition of the present application can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present application has a synergistic effect with other therapeutic drugs, the dose thereof should be adjusted according to the actual situation.

[0030] Beneficial technical effects: The present application adopts a method for evaluating the therapeutic effect of monomer compounds on H22 cell-induced hepatocellular carcinoma nude mice by subcutaneous injection. The experiment proves that the polycyclic polyisoprenyl phloroglucinol compound has a significant effect on reducing the weight of tumor tissue and improving liver damage in hepatocellular carcinoma nude mice at a dose of 25 mg / kg and 50 mg / kg. The hepatocellular carcinoma drug prepared therefrom can reduce purine metabolism and inhibit the MAPK pathway, effectively reduce the tumor load, repair damaged hepatocytes, and significantly improve the liver damage of tumor-bearing mice.

[0031] For better understanding and implementation, the present application is described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 Comprehensive evaluation results of the nude mouse model, body weight and tumor growth characteristics of the mice in Example 2;

[0033] Figure 2 Comprehensive evaluation results of the liver tissue morphological changes, sinusoidal blood flow and serum AST / ALT activity of the mice in Example 2;

[0034] Figure 3 Evaluation results of the purine metabolism level of the mice in Example 2. DETAILED DESCRIPTION

[0035] The essential content of the present application is further illustrated below with reference to the examples, but the protection scope of the present application is not limited thereto. Although the present application is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can be modified or replaced equivalently without departing from the essence and scope of the technical solutions of the present application.

[0036] The chemical structural formula of the polycyclic polyisoprenyl depsidate compound Attenuatumione D in the embodiments of the present application is shown as follows:

[0037]

[0038] Example 1: Preparation of Attenuatumione D

[0039] S1: The gold coin grass powder is mixed with 8 times the volume of 95% ethanol, heated to reflux extraction for 1 h, extracted for 3 times, filtered, and the 3 times of extraction solutions are combined and concentrated under reduced pressure to a thick solution to obtain the total extract of gold coin grass;

[0040] S2: The total extract of gold coin grass is passed through a D101 macroporous resin column, and ethanol and water are used as eluents to perform gradient elution from 80% ethanol to 100% ethanol, the eluate of the 100% ethanol section is collected, concentrated by rotary evaporation, and the solvent is removed to obtain a mixture A after concentration;

[0041] S3: The mixture A is separated by preparative HPLC with Rp-C18 reversed-phase silica gel filler, wherein the mobile phase is acetonitrile with a concentration of 80%, the flow rate is 8.0 ml / min, and the peak with a retention time of 18.6 min is collected, concentrated by rotary evaporation, and the solvent is removed to obtain the polycyclic polyisoprenyl depsidate compound Attenuatumione D.

[0042] The mixture A is prepared by preparative HPLC using a RP-C18 column (5 μm, 250 x 20 mm) eluted with acetonitrile / water 80:20 by volume by volume at a flow rate of 8 mL / min and detected at 205 nm wavelength, to obtain finally the polycyclic polyprenyl depsidate compound of the application Attenuatumione D.

[0043] Example 2: Intervention of Attenuatumione D on H22 cell-induced hepatocellular carcinoma in nude mice by subcutaneous injection Pharmacological evaluation of the effect

[0044] 1. Experimental samples

[0045] Preparation of the solution of the sample to be tested: the polycyclic polyprenyl depsidate compound of the application Attenuatumione D was prepared as described in Example 1. An appropriate amount of sample was accurately weighed and dissolved in DMSO to obtain a stock solution at 10 mM, to be used in the pharmacological tests.

[0046] 2. Experimental subjects

[0047] BALB / c nude mice (male, SPF grade, 7-8 weeks old) weighing 20 ± 2 g were used. The mice were acclimatized in a sterile environment for three days before the experiment.

[0048] 3. Experimental method

[0049] 3.1. Establishment of the hepatocellular carcinoma nude mouse model

[0050] The BALB / c nude mice were acclimatized in a sterile environment (temperature 22 ± 2°C, humidity 60% ± 5%, 12 hours light / 12 hours dark cycle) for three days before the experiment. All operations were carried out in strict accordance with the "Guidelines for the Management of Laboratory Animals" and the relevant regulations of the People's Republic of China, while referring to GB / T 35892-2018 "Guidelines for the Welfare and Ethical Review of Laboratory Animals". 1 x 10 7 H22 tumor cells were transplanted subcutaneously into the mesentery of the nude mice in a volume of 0.2 mL, ensuring that the required number of cells for the experiment was sufficient.

[0051] 3.2. Administration method and tissue sampling

[0052] Twenty male BALB / c nude mice were randomly divided into four groups (n = 5 for each group) 24 hours after the subcutaneous injection of H22 cells in the previous step, and intravenous administration was performed daily for 16 consecutive days. The groups were as follows:

[0053] Group I, type (saline); group II, CTX (25 mg / kg BW); group III, Attenuatumione D (25 mg / kg BW); group IV, Attenuatumione D (50 mg / kg BW).

[0054] The blank group was physiological saline, the positive control group was cyclophosphamide (CTX), and the administration groups were 25 mg / kg / day and 25 mg / kg / day. The dose of the administration groups was selected according to the pre-experiment, which showed no toxic effect on nude mice. On the last day, all nude mice were weighed and sacrificed, and then the tumors were removed and weighed.

[0055] 3.3 Inhibition rate of tumor

[0056] The anti-tumor effect was expressed as the inhibition rate of tumor (%), and the calculation formula was: inhibition rate (%) = [model group (tumor weight; g) - treatment group (tumor weight; g) / model group (tumor weight; g)] x 100.

[0057] 3.4 Histological analysis

[0058] The liver tissues of each group were fixed with formalin (10% neutral buffer), dehydrated with gradient ethanol, transparentized with xylene, and embedded with paraffin. The tumor tissues were cut from the middle and cut into 4 μm thick sections, which were stained with hematoxylin and eosin (H&E) and observed under a microscope.

[0059] 3.5 Serum metabolomics analysis

[0060] 100 mL of serum sample was taken and 300 mL of methanol was added for extraction. After incubation in ice bath for 10 minutes, centrifugation was performed at 4°C (13,000 rpm) for 10 minutes. The supernatant was freeze-dried and resuspended with 50 mL of methanol / water (3:1 by volume). After centrifugation at 4°C (13,000 rpm) for 10 minutes again, the supernatant was collected for analysis. To ensure the consistency of the experimental conditions and the quality of the data, all extracted tissues or serum samples were mixed to prepare a quality control sample (QC) for serum metabolomic analysis. The pretreatment process of the quality control sample was synchronized with the experimental samples.

[0061] A Vanquish Ultra Performance Liquid Chromatography system (Thermo Fisher Scientific, Waltham, USA) coupled with an Orbitrap Exploris 120 mass spectrometer was used. The Hypersil GOLD C18 column (100 mm x 2.1 mm, 1.9 mm, Thermo Fisher Scientific) was used for analysis. The mobile phase A was water containing 0.1% formic acid, and the mobile phase B was acetonitrile containing 0.1% formic acid. The gradient elution program was set as follows: 0-5 min 5% B, 0.5-20 min 5%-100% B, 20-27.9 min 100% B, 28-30 min 5% B. The flow rate was 0.30 mL / min, and the column oven was set at 40 °C. The injection volume was 5 mL for each analysis, and the sample order was randomly arranged. The mass spectrometry scan range was set between 50 and 1000 m / z, and the ion spray voltage was set at 3200 V in the electrospray ionization mode, with the vaporizer temperature maintained at 350 °C. In the electrospray ionization mode, the hydrogen-deuterium collision energy (%) was set at 30, 60, or 90, respectively. The temperature was maintained at 4 °C during the analysis of all samples.

[0062] 4. Analysis of experimental results

[0063] 4.1 Anti-tumor effect of Attenuatumione D in vivo

[0064] As shown in Figure 1, to fully evaluate the anti-tumor activity of Attenuatumione D, a H22 hepatocellular carcinoma nude mouse model was established (A), the body weight of nude mice (B), the morphological changes of tumors (C), and the tumor weight (D) were determined. Figure 1 Figure 1 Figure 1 Figure 1 Figure 1

[0065] 4.2 Body weight

[0066] To evaluate the effect of Attenuatumione D on the growth status and body weight of mice, we studied the drug-induced toxicity as an indicator. As shown in Figure 2, the body weight of the mice in the control group increased gradually, while the body weight of the mice in the treatment group decreased significantly, indicating that Attenuatumione D had a significant effect on the growth status and body weight of the mice. Figure 1 ​​​​​As shown in Figure B, the initial body weights of all nude mice in this study were similar, with no significant difference (p>0.05). However, the final body weight of the model group (33.98±1.47g) was significantly higher than that of the CTX-treated group (30.60±1.17g), indicating that CTX has significant side effects. Furthermore, the physiological state of the mice in the Attenuatumione D-treated group was better than that of the model group and the CTX group, exhibiting smooth skin, normal growth, frequent activity, and rapid response. Meanwhile, the initial and final weight gain rates of the Attenuatumione D-treated group (250 or 500 mg / g BW; 1.40±0.06 or 1.42±0.10) were not significantly different from those of the model group (1.39±0.06), indicating that the mice in this treatment group did not exhibit the aforementioned side effects and toxicity signs.

[0067] 4.3 Histopathological Analysis

[0068] like Figure 2 As shown, this study used hematoxylin-eosin (H&E) staining to observe morphological changes in liver tissue under a microscope. Figure 2 As shown in Figure A, the hepatocytes in the control group were neatly arranged, exhibiting a typical clustered structure, and there was clear evidence of blood flow through the sinusoidal spaces formed by the central vein. Figure 2 (B). However, compared with the control group, the liver tissue of the model group (HCC-induced tumor-bearing mice) showed features such as lymphoma proliferation, hepatic vascular congestion, and hepatocyte atypia and degeneration. Figure 2 In HCC, not only does portal vein congestion occur, but the volume and number of lymphoid tissue surrounding the vein also increase. Furthermore, the dilation and congestion of the central vein indicate impaired blood flow in this area, possibly due to the accumulation of atypical hepatocytes and lymphoid tissue. Some hepatocytes exhibit ballooning degeneration and microfatty degeneration, hallmarks of HCC-induced cell damage. H&E staining of Attenuatumione D group mice treated with HCC (+50 mg / kg) showed mild mononuclear inflammatory aggregation, with only slight swelling and vacuolar degeneration of hepatocytes. In particular, the Attenuatumione D group significantly reduced hepatic vascular congestion, decreased hepatocyte microfatty degeneration, and enhanced macrophage activity. This study suggests that the presence of macrophages indicates that the liver is responding to inflammation or damage and actively clearing toxins and other harmful substances from the body. Figure 2C and D). In addition, the liver sections of HCC + 50mg / kg attenuatumione D group mice showed that the hepatocytes were arranged in cord-like structures, relatively intact, which indicated that their functions were gradually restored to normal levels. The experimental results showed that the 50mg / kg dose of attenuatumione D treatment not only improved the cell histological features, but also minimized the degree of liver damage such as coagulopathy and vacuolar degeneration, and was more effective in improving liver histology.

[0069] 4.4 Attenuatumione D reversed the abnormal purine metabolism in serum of HCC nude mice

[0070] Metabolomics analysis showed that there were significant changes in three metabolites Figure 3 ). The most significant differential metabolites in the drug administration group compared to the model group were Inosine, Guanine and Hypoxanthine in purine metabolism. Abnormal purine metabolism promotes chronic inflammatory response and plays a key role in various biological processes of cancer cells. Abnormal purine metabolism has been found in various cancers including HCC and is considered an important mechanism for increasing cancer risk. Studies have shown that the abnormality of purine metabolism in the model group is increased, and after administration, the level of purine metabolism is significantly reduced, and the metabolite level of the drug administration group at a dose of 50mg / kg / day of attenuatumione D is close to that of the positive drug. The above research results show that attenuatumione D can significantly reverse the abnormal purine metabolism of hepatocellular carcinoma nude mice to normal levels.

[0071] The above results show that the polycyclic polyisoprenyl phloroglucinol compound attenuatumione D of the present application can significantly reduce the tumor weight of hepatocellular carcinoma nude mice at a dose of 25mg / kg and 50mg / kg, reverse the abnormal purine metabolism of hepatocellular carcinoma nude mice, and can be used for preparing a therapeutic drug for hepatocellular carcinoma.

[0072] The above-described embodiments only express several embodiments of the present application, which are described in more detail and in detail, but should not be understood as limiting the scope of the patent. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, and the present application also intends to include these modifications and improvements.

Claims

1. The use of polycyclic polyisoprenyl depsidate attenuatumione D in the preparation of a drug for treating hepatocellular carcinoma, characterized in that: The chemical structural formula of the polycyclic polyisoprenyl depsidate Attenuatumione D is as follows: ​ 2. Use according to claim 1, characterized in that: The content of the polycyclic polyisoprenyl depsidate Attenuatumione D in the medicine is 0.1-95wt%.

3. Use according to claim 1, characterized in that: The polycyclic polyisoprenyl depsidate Attenuatumione D is extracted and separated from the Hypericum attenuatum.

4. The use according to claim 1, characterized in that: The preparation method of the polycyclic polyisoprenyl depsidate Attenuatumione D comprises the following steps: S1: the Hypericum attenuatum powder is mixed with 8 times of 95% ethanol, heated to reflux extraction for 1h, extracted for 3 times, filtered, and the 3 times of extraction solutions are combined, concentrated under reduced pressure to a thick solution, to obtain Hypericum attenuatum total extract; S2: the Hypericum attenuatum total extract is passed through a D101 macroporous resin column, ethanol and water are used as eluents, gradient elution is carried out from 80% ethanol to 100% ethanol, the eluent of the 100% ethanol section is collected, concentrated by rotary evaporation, and the solvent is removed, to obtain a mixture A after concentration; S3: the mixture A is separated by preparative HPLC of Rp-C18 reverse-phase silica gel filler, wherein the mobile phase is 80% acetonitrile, the flow rate is 8.0ml / min, the peak with a retention time of 18.6min is collected, concentrated by rotary evaporation, and the solvent is removed, to obtain the polycyclic polyisoprenyl depsidate Attenuatumione D.

5. The use according to claim 1, characterized in that: The medicine comprises a pharmaceutically acceptable excipient.

6. The use according to claim 1, characterized in that: The medicine is any one of a liquid preparation, a solid preparation, a semi-solid preparation, and a gaseous preparation.

7. The use according to claim 1, characterized in that: The dosage form of the medicine is any one of an oral liquid, a tablet, a capsule, a dripping pill, a granule, or an injection.

8. Use according to any one of claims 1 to 7, characterized in that: The effective administration dose of the medicine is 25-50mg / kg / day.

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