Application of manganese-type superoxide dismutase with high stability in prevention and treatment of prostatitis

Abstract

The application belongs to the field of biological medicine, and relates to application of a manganese-type high-stability superoxide dismutase with an amino acid sequence as shown in SEQ ID NO: 1 in prevention and treatment of prostatitis. The manganese-type high-stability superoxide dismutase can reduce the symptoms of CP / CPPS, and has good application value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to the prevention and treatment of prostatitis. BACKGROUND

[0002] Prostatitis is a common clinical disease in urology, which can be divided into: acute bacterial prostatitis (type I), chronic bacterial prostatitis (type II), chronic prostatitis / chronic pelvic pain syndrome (CP / CPPS) (type III) and asymptomatic prostatitis (type IV). Type III can be further divided into inflammatory type III (a) and inflammatory type III (b). CP / CPPS has the highest incidence among the four types, accounting for about 90-95% of patients with prostatitis, and is the most common urological disease in men under 50 years old. The main symptoms of CP / CPPS include long-term pain in the perineum and lower abdomen, abnormal urination and neuropsychiatric symptoms such as sexual dysfunction and decreased fertility, etc. Clinically, CP / CPPS has a slow onset, stubborn symptoms, repeated attacks and is difficult to cure, which seriously affects the quality of life of patients. At present, the treatment of CP / CPPS mainly focuses on relieving symptoms and cannot completely cure it. The pathogenesis and pathophysiology of CP / CPPS are not yet clear, which may be related to oxidative stress, pathogenic bacterial infection and dysfunction of the nervous, immune and endocrine systems. Although many studies have found that SOD can remove excess ROS in tissues, the treatment effect of ordinary SOD is still not ideal due to poor stability. SUMMARY

[0003] The present application applicant found in the research that manganese-type high-stability superoxide dismutase MS-SOD has a good therapeutic effect on prostatitis, therefore,

[0004] In the first aspect of the present application, the use of MS-SOD in the preparation of a drug for preventing or treating prostatitis is provided, wherein the amino acid sequence of the manganese-type high-stability superoxide dismutase is shown in SEQ ID NO: 1.

[0005] Preferably, the drug is administered by oral or injection route; further preferably, the injection is intravenous injection or intraperitoneal injection.

[0006] Preferably, the dosage form of the drug is tablet, capsule, powder, powder injection, injection or aerosol.

[0007] Preferably, the drug further comprises one or more pharmaceutically acceptable excipients, such as diluents, binders, wetting agents, disintegrants, solvents and / or buffers, etc. These excipients are prepared into drugs with MS-SOD in a manner known in the art, and the amount thereof is known to those skilled in the art.

[0008] In another aspect, the present application provides a method for preventing or treating prostatitis, comprising administering MS-SOD to an individual in need thereof.

[0009] Preferably, the prostatitis according to the present application is chronic prostatitis / chronic pelvic pain syndrome.

[0010] The MS-SOD according to the present application can alleviate the CP / CPPS symptoms of a CP / CPPS rat model, and has good stability and activity under high temperature, high pH, and the action of proteases such as pepsin and trypsin, and has good application value. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 A is the result of prostate index of each group;

[0012] Figure 1 B~1F respectively show the pain threshold results of rats in each group at different times;

[0013] Figure 2 A is the result of hematoxylin-eosin staining of histopathology;

[0014] Figure 2 B is the result of Masson staining of histopathology;

[0015] Figure 3 A~3C are the results of immunohistochemical (IHC) staining of CD3, CD45 and Mlp1a cells; Figure 3 D~3F are the expression value graphs of CD3, CD45 and MIP1a;

[0016] Figure 4 A is the fluorescence graph of ROS content; Figure 4 B is the relative numerical result graph of ROS;

[0017] Figure 4 C~4G are the results of detection of homogenate oxidative markers;

[0018] Figure 5 A~5D represent the results of TNF-α, IL-1β, IL-8 and CRP levels measured by ELISA method;

[0019] Figure 5 E~5G are the RT-qPCR results of TNF-α, IL-1β, IL-8;

[0020] Figure 6 A~6E are the results of Western blotting and numerical conversion. DETAILED DESCRIPTION

[0021] The present application will be described in detail below through specific embodiments, but the content of the present application is not limited thereto.

[0022] In the following examples, the reagents and instruments used are conventional reagents and instruments in the art, which can be obtained by commercial means, unless otherwise specified; the experimental methods used are conventional methods in the art, and those skilled in the art can undoubtedly implement the schemes and obtain the corresponding results according to the content of the examples.

[0023] The MS-SOD sequence used in the following examples is shown as SEQ ID NO: 1, and its preparation process is described in detail in Example 1 of CN105624126A, which is incorporated herein by reference.

[0024] The sequence of MS-SOD is:

[0025] MPYPFKLPDLGYPYEALEPHIDAKTMEIHHQKHHGAYVTNLNAALEKYPYLHGVEVEVLLRHLAALPQDIQTAVRNNGGGHLNHSLFWRLLTPGGAKEPVGELKKAIDEQFGGFQALKEKLTQAAMGRFGSGWAWLVKDPFGKLHVLSTPNQDNPVMEGFTPIVGIDVWEHAYYLKYQNRRADYLQAIWNVLNWDVAEEFFNKA (SEQ ID NO: 1).

[0026] Experimental reagents:

[0027] Freund's complete adjuvant CFA: Sigma, USA;

[0028] SOD, CAT, GSH-PX and MDA detection kit: Nanjing Jiancheng Biological Technology Institute;

[0029] NO detection kit: Shanghai Biyun Tian Biological Technology Co., Ltd.;

[0030] DCFH-DA ROS detection kit; CRP ELISA kit: Beijing Bai'elai Boluo Technology Co., Ltd.;

[0031] IL-1β and TNF-α ELISA kit: R&D Systems, USA;

[0032] IL-8 Elisa kit: Shanghai Xitang Biological Technology Co., Ltd.;

[0033] Masson's trichrome staining kit: Jiangsu Keygen Biotechnology Co., Ltd.;

[0034] One-step immunohistochemical kit (rabbit and mouse universal): Jiangsu Keygen Biotechnology Co., Ltd.;

[0035] Anti-CD3, anti-CD45 and anti-GAPDH antibodies: Abcam (UK);

[0036] Anti-MIP1a antibody: Mybiosource, USA;

[0037] Anti-NF-κB P65, anti-phosphorylated NF-κB P65, anti-P38-MAPK and anti-phosphorylated P38-MAPK antibodies: Cell Signaling Technology, USA;

[0038] Total RNA reagent kit: Tiangen Biotech Co., Ltd., China;

[0039] RT-PCR kit: Bio-Rad Life Sciences (Shanghai) Co., Ltd.;

[0040] H&E staining kit: Jiangsu Kaiji Biotechnology Co., Ltd.

[0041] Laboratory animals:

[0042] Forty male Sprague-Dawley rats were purchased from Shanghai Ruijin Biotechnology Co., Ltd. and allowed to rest for one week. The rats were housed in an environment of 25±3℃, exposed to alternating cycles of 12 hours of light and 12 hours of darkness, and had free access to food and water.

[0043] Experimental methods

[0044] Forty SD rats were randomly divided into 5 groups, 8 rats in each group: blank sham operation group (control group), model group, MS-SOD low dose (100 u / g), MS-SOD medium dose (300 u / g) and MS-SOD high dose (500 u / g) group. According to the method reported by Peng et al., CP / CPPS was induced, which is one of the most commonly used CP / CPPS induction models; first, the rats were anesthetized with 10% chloral hydrate (1 ml / 300 g) intraperitoneally, then the abdominal hair was scraped, the skin was disinfected, and the lower abdomen was incised at the midline to expose the prostate. After injecting 0.1 mL of CFA into the prostate of the model group and MS-SOD high / medium / low dose group rats, the incision was layered sutured. The sham operation group was only turned over the prostate several times after laparotomy, and then the abdomen was sutured. Two days after the modeling was successful, the model group and the MS-SOD group were orally administered with normal saline 1 ml or MS-SOD for 4 weeks, and the sham operation group was not given any treatment. On day 28, blood was collected from the abdominal aorta under 10% chloral hydrate anesthesia. After centrifugation at 2000 x g for 15 minutes, the supernatant was collected and stored at -80°C for subsequent biochemical tests. At the same time, the prostate was quickly removed, washed with normal saline, and the adipose tissue and adherent blood were removed. The wet weight of the prostate was recorded, a part of the prostate was taken for histological imaging, and another part was stored at -80°C for biochemical analysis. Finally, at the end of the experiment, the rats were euthanized by inhaling excess isoflurane. This study was conducted in accordance with the guidelines of the "Helsinki Declaration".

[0045] Statistical analysis

[0046] Statistical analysis was performed using SPSS software. All data are expressed as mean ± standard deviation (SD), using (x ± s), where N represents the number of animals. Normal distribution data were analyzed by t-test. One-way ANOVA and Tukey's test were used to evaluate the statistical significance of differences between groups. *P <0.05, #P <0.05, **P <0.01 and ##P <0.01 were statistically significant.

[0047] Example 1: Basic evaluation

[0048] Prostate index determination

[0049] On day 28 after surgery, the body weight, total weight of the abdomen and the lateral side of the rats in each group were recorded after anesthesia with 10% chloral hydrate to calculate the prostate index: Prostate index = Prostate weight (mg) / Rat weight (g) x 100%.

[0050] Results: The prostate index (PI) of the model group rats was significantly higher than that of the sham operation group (**P <0.01), and the PI of the MS-SOD (300 U / g, 500 U / g) group was significantly lower than that of the model group (#P <0.05 and #P <0.01, respectively) Figure 1A). This indicates that MS-SOD can reduce the prostate index of CP / CPPS.

[0051] Pain threshold assessment

[0052] Following the methods of Cheng et al. and Ni et al., pain threshold tests were performed on rats 1 day before surgery and 7, 14, and 28 days after surgery. The rat scrotum (BME-404) was vertically stimulated with a Von-Frey filament five times consecutively, each stimulation lasting at least 10 seconds. Violent abdominal contractions, scratching, jumping, and licking at the stimulated area were used as the pain threshold, and the average values ​​were recorded.

[0053] result: Figure 1 B shows the changes in pain threshold among the rat groups throughout the 28-day experiment. It was found that the pain threshold of the rats in each group gradually increased to a certain level. Importantly, the pain threshold of the rats in each group was measured before the experiment, and no significant initial differences were found between the groups. Figure 1 C). On day 7, a significant decrease in pain threshold was observed in the model group compared to the sham surgery group (**P<0.01), while there were no significant differences between the other groups and the model group (P>0.05). Figure 1 D). On day 14, the pain threshold of rats in the model group was significantly lower than that in the sham-operated group (**P<0.01), and the pain threshold of the model group was significantly higher than that in the high-dose MS-SOD group (500U / g##P<0.01); Figure 1 E). On day 28, compared with the sham-operated group, the pain threshold of rats in the model group was significantly reduced (**P<0.01), and MS-SOD (300U / g, 500U / g) significantly increased the pain threshold (##P<0.01). Figure 1 F). This indicates that MS-SOD can increase the pain threshold of CP / CPPS.

[0054] Evaluation of serum pro-inflammatory mediators

[0055] Serum samples were collected, and the levels of IL-1β, IL-8, TNF-α, and CRP were measured by ELISA to analyze the therapeutic effect of MS-SOD on rat CP / CPPS.

[0056] Result: As Figure 5 As shown in the AD, compared with the sham surgery group, the levels of TNF-α, IL-8, IL-1β and CRP in the model group were significantly increased (**P<0.01), while the levels in the high, medium and low dose MS-SOD groups were significantly decreased compared with the model group (P<0.05 or P<0.01), suggesting that MS-SOD has an anti-inflammatory effect.

[0057] Example 2: Histopathological Experiment

[0058] H&E staining (hematoxylin and eosin staining)

[0059] Prostate tissues were first fixed with 4% paraformaldehyde for 24 hours, then paraffin-embedded. The sections were 5 pm thick, and H&E staining was used for prostate tissue morphological examination. Briefly, according to the instructions of the H&E staining kit, the sections were deparaffinated, stained, dehydrated, and sealed. Representative fields were selected under an optical microscope (400x magnification) for imaging.

[0060] Results: H&E staining was used to observe the changes in prostate tissue structure in each group and to evaluate the therapeutic effect of MS-SOD on CP / CPPS. As shown in Figure 2 As shown in Fig. A, no edema was observed in the prostate stroma of the sham operation group, and the gland lumen structure was complete, with no obvious hyperplasia of acinar epithelium. No inflammatory cell infiltration was observed in all structures. Edema, increased secretion, and vascular hyperemia were observed in the model group, and the prostate structure was obviously abnormal, such as abnormal cavity secretion and uneven tissue hyperplasia. Notably, the MS-SOD (100, 300, and 500 u / g) groups showed different degrees of gland inflammation, and the cavity discharge decreased in a dose-dependent manner.

[0061] Masson staining:

[0062] After fixation, paraffin embedding, and sectioning of the prostate tissue, a Masson trichrome staining kit was used to observe the fibrosis of the prostate tissue. Briefly, according to the instructions, the sections were deparaffinated, stained, rinsed, dehydrated, dried, and sealed, respectively. Images were observed under an optical microscope (400x magnification), and representative fields were selected for imaging.

[0063] Results: Masson staining was used to observe the degree of fibrosis in the prostate tissue of each group and to evaluate the therapeutic effect of MS-SOD on fibrous connective tissue during CP / CPPS. As shown in Figure 2 As shown in Fig. B, no obvious fibrosis was observed in the sham operation group. The model group showed a high degree of liver fibrosis, which was widely distributed. The MS-SOD (100, 300, and 500 u / g) groups showed different degrees of fibrosis, which was scattered, and the degree of fibrosis decreased in a dose-dependent manner as the concentration of MS-SOD increased.

[0064] Example 3: Immunohistochemical examination

[0065] The paraffin-embedded sections of each group were placed in an oven and baked for 3 hours. After drying, routine xylene deparaffination was performed, followed by graded alcoholization of the slides, and then hydration. The slides were incubated with 3% H2O2 at room temperature for 10 min using citric acid antigen retrieval solution to inactivate endogenous enzymes.

[0066] One-step immunohistochemistry kit (rabbit, mouse universal) was used for the experiment. Reagent A (normal goat serum) was incubated for 30 minutes, and PBS was washed for 3 times, each for 2 minutes. The diluted antibody (CD3 antibody [1:50], CD45 antibody [1:200] and Mlp1a antibody [1:100]) was added to the slide, and after incubation at 4°C overnight, PBS was washed for 3 times, each for 2 minutes. Next, reagent B (polymer enhancer) was added for 30 minutes, and then PBS was washed for 3 times, each for 2 minutes. Reagent C (HRP-labeled anti-mouse rabbit polymer) was added for 30 minutes, and then PBS was washed for 3 times, each for 2 minutes. Finally, 3,3'-diaminobenzidine (DAB) was added for color development for 3-5 minutes, and the reaction time was controlled under a microscope. After washing with water, hematoxylin solution was added for staining. After the slice was dried, neutral gum was used for fixation. Two experienced researchers conducted blind evaluation of IHC readings. Each IHC slice was observed and photographed under a microscope at 400 times magnification, and semi-quantitative analysis was performed by integral optical density analysis.

[0067] Results: In order to preliminarily distinguish the types of inflammatory cells infiltrating the prostate tissue during CP / CPPS, immunohistochemical (IHC) staining of CD3, CD45 and Mlp1a cells was performed Figure 3 A-C). CD3 is a biomarker of T lymphocytes, CD45 is a non-specific marker of leukocytes, and Mlp1a is a biomarker of macrophages. As shown in Figure 3 D-F, the levels of CD3, CD45 and MIP1A in the model group were significantly higher than those in the sham operation group (**P<0.01). Interestingly, compared with the model group, the expression of CD3, CD45 and MIP1A in the MS-SOD (100, 300 and 500 U / g) group was significantly reduced (P<0.05 or P<0.01), and it was reduced in a dose-dependent manner with the increase of MS-SOD concentration.

[0068] Example 4: Reactive oxygen species fluorescent probe

[0069] ROS content was measured using DCFH-DA. Each group of prostate specimens was placed in 4% paraformaldehyde fixative for 24 hours, then transferred to 30% sucrose for 48 hours. The tissue blocks were placed in small boxes, and a suitable amount of OCT embedding agent was added to soak the tissue; then the box was slowly placed in a small cup containing liquid nitrogen. Once the bottom of the box was in contact with the liquid nitrogen, it was left in place rather than being immersed in the liquid nitrogen. The tissue was quickly frozen (about 10 seconds). After the frozen block was ready, the sample was sectioned in a cryostat. Next, after the clean working solution was applied to the surface of the section at room temperature for 10 minutes, it was carefully removed. Finally, DCFH-DA staining solution was added to the frozen section, and incubation was continued at 37°C for 20 minutes, followed by fluorescence microscopy observation and photography (630x; green: DCFH-DA, blue: DAPI).

[0070] Oxidative stress is the result of an imbalance between ROS production and endogenous antioxidant capacity, either due to excessive ROS production or decreased clearance capacity. Therefore, this example used the dichlorofluorescein diacetate (DCFH-DA) probe to quantitatively analyze ROS in prostate tissue using the CP / CPPS model. As shown in Figure 4 As shown in A and B, the ROS level of the model group was significantly higher than that of the sham operation group (**P < 0.01), but compared with the model group, the ROS level of the MS-SOD (100, 300 and 500 U / g) group was significantly reduced (P < 0.01). This indicates that MS-SOD can reduce oxidative damage in a dose-dependent manner.

[0071] Example 5: Homogenate detection

[0072] Homogenate preparation

[0073] The prostate samples were weighed and divided into two parts before adding sterile normal saline (for ELISA) or RIPA lysate (for western blotting). The tissue homogenate was prepared with a tissue homogenizer at 4°C, centrifuged at 10,000 x g for 10 minutes, and the supernatant was collected and stored at -20°C.

[0074] Determination of oxidative stress level

[0075] According to the instructions of the ELISA kit, a suitable amount of sample homogenate was detected for antioxidant markers, including malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) levels, for quantitative analysis to evaluate the antioxidant capacity of prostate tissue.

[0076] As shown in Figure 4C-G showed that compared with the sham operation group, the levels of SOD, GSH-Px and CAT in the model group were significantly reduced, and the levels of MDA and NO were significantly increased (**P <0.01). Compared with the model group, the levels of SOD, GSH-Px and CAT in the MS-SOD (100, 300 and 500 u / g) group were significantly increased, and the levels of MDA and NO were significantly reduced (P <0.05 or P <0.01), and showed a dose-dependent manner. It is suggested that MS-SOD has the effect of enhancing free radical scavenging and reducing peroxidation.

[0077] RT-qPCR analysis

[0078] The expression of pro-inflammatory mediators (IL-1β, TNF-α, IL-8) was detected by LightCycler 480 real-time fluorescence quantitative PCR system.

[0079] In order to determine the effect of MS-SOD on inflammatory cytokines, the mRNA levels of IL-1β, TNF-α and IL-8 in prostate were detected by quantitative real-time polymerase chain reaction (RT-qPCR) using LightCycler 480 real-time fluorescence quantitative PCR system. As shown in Figure 5 E-G showed that compared with the sham operation group, the expression of IL-1β, TNF-α and IL-8 in the model group was significantly increased (**P <0.01). Compared with the model group, MS-SOD (100, 300 and 500 u / g) significantly reduced the expression of these cytokines (P <0.01).

[0080] Western blot

[0081] The protein concentration of a small part of prostate tissue homogenate was measured according to the instructions of BCA kit (Sigma). The tissue samples and protein markers were electrophoresed by SDS-PAGE, and the separated polypeptides were transferred to PVDF membrane. The membrane was soaked in Tris buffer (TBS), moved to blocking buffer (5% skim milk powder and 0.1% Tween [TBST] solution in TBS), then washed with TBST solution, and finally incubated with monoclonal phosphorylated p65 antibody, p65, phospho-p38 and p38 antibody at 4°C overnight. The next day, the cell membrane was washed with TBST three times, incubated with goat anti-rabbit or mouse IgG antibody at room temperature for 1 h. The membrane was washed with TBST three times, and ECL chemiluminescence reagent was added to observe protein expression. The images were collected on Tanon6600 imaging workstation, and the optical density analysis was performed by imageproplus6.0.

[0082] It is well known that the transcription of inflammatory cytokines (IL-1β, TNF-α and IL-8) depends on the activation of nuclear factor NF-κB, and p38 mitogen-activated protein kinase (MAPK) is an important signal molecule for intracellular signal transduction, both of which play an important role in the regulation of inflammatory response and oxidative stress, so it is speculated that MS-SOD can regulate the progression of CP / CPPS in rats by inhibiting the NF-κB and p38mapk signaling pathways. Therefore, we detected the expression of phosphorylated NF-κB p65 and p38MAPK proteins in prostate tissue. As Figure 6 As shown in A, C, E, the phosphorylation levels of p65 and p38 in the model group were significantly higher than those in the sham operation group (**P<0.01), and the phosphorylation levels of p65 and p38 in the MS-SOD (100, 300, 500U / g) group were significantly lower than those in the model group (##P<0.01). Figure 6 B, D respectively indicate that the amount of unphosphorylated p65 and p38 in the model group and the SOD group has no significant change. It is suggested that MS-SOD may reduce inflammatory response and oxidative stress damage by inhibiting the NF-κB, p65 and p38mapk signaling pathways.

[0083] From the above experimental results, we can see that:

[0084] (1) MS-SOD can regulate the inflammatory response and oxidative stress of CP / CPPS rat model;

[0085] (2) NF-κb p65 and p38mapk signaling pathways play a key role in MS-SOD regulating the inflammatory response and oxidative stress damage of CP / CPPS rats. These results suggest that MS-SOD is a potential candidate for the treatment of CP / CPPS. SEQUENCE LISTING <110> Hangzhou Ruida Pharmaceutical Technology Co., Ltd. Kailaihengji Biological Medicine (Hangzhou) Co., Ltd. <120> Application of manganese-type high-stability superoxide dismutase in prevention and treatment of prostatitis <130> DSP1F223341JW <150> CN202110910916.0 <151> 10 August 2021 <160> 1 <170> SIPOSequenceListing 1.0 <210> 1 <211> 204 <212> PRT <213> Artificial Sequence <400> 1 Met Pro Tyr Pro Phe Lys Leu Pro Asp Leu Gly Tyr Pro Tyr Glu Ala 1 5 10 15 Leu Glu Pro His Ile Asp Ala Lys Thr Met Glu Ile His His Gln Lys 20 25 30 His His Gly Ala Tyr Val Thr Asn Leu Asn Ala Ala Leu Glu Lys Tyr 35 40 45 Pro Tyr Leu His Gly Val Glu Val Glu Val Leu Leu Arg His Leu Ala 50 55 60 Ala Leu Pro Gln Asp Ile Gln Thr Ala Val Arg Asn Asn Gly Gly Gly 65 70 75 80 His Leu Asn His Ser Leu Phe Trp Arg Leu Leu Thr Pro Gly Gly Ala 85 90 95 Lys Glu Pro Val Gly Glu Leu Lys Lys Ala Ile Asp Glu Gin Phe Gly 100 105 110 Gly Phe Gin Ala Leu Lys Glu Lys Leu Thr Gin Ala Ala Met Gly Arg 115 120 125 Phe Gly Ser Gly Trp Ala Trp Leu Val Lys Asp Pro Phe Gly Lys Leu 130 135 140 His Val Leu Ser Thr Pro Asn Gln Asp Asn Pro Val Met Glu Gly Phe 145 150 155 160 Thr Pro Ile Val Gly Ile Asp Val Trp Glu His Ala Tyr Tyr Leu Lys 165 170 175 Tyr Gln Asn Arg Arg Ala Asp Tyr Leu Gln Ala Ile Trp Asn Val Leu 180 185 190 Asn Trp Asp Val Ala Glu Glu Phe Phe Asn Lys Ala 195 200

Claims

1. Use of a manganese-type hyperstable superoxide dismutase in the preparation of a medicament for the treatment of prostatitis, wherein, The amino acid sequence of the manganese-type high-stability superoxide dismutase is shown as SEQ ID NO: 1; The medicine is administered through oral or injection route. The prostatitis is chronic prostatitis / chronic pelvic pain syndrome.

2. The use according to claim 1, wherein, The injection is intravenous injection or intraperitoneal injection.

3. The use according to claim 1, wherein, The dosage form of the medicine is tablet, capsule, powder, powder injection, injection or aerosol.

4. The use according to any one of claims 1 to 3, wherein The medicine further comprises one or more pharmaceutically acceptable adjuvants.

5. Use according to claim 4, wherein, The adjuvant is diluent, binder, wetting agent, disintegrant, solvent and / or buffer.

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