阅读说明：本技术 用于预防反复性泌尿道感染的药学组合物 (Pharmaceutical composition for preventing recurrent urinary tract infections ) 是由 王雅俊 张玉青 马晓恬 陈振耀 李苑如 蒙恩 张尚仁 于 2020-04-24 设计创作，主要内容包括：本发明提供了一种戊聚醣多硫酸钠(pentosan polysulfate sodium；PPS)用于制备预防人类个体中反复性泌尿道感染的药物的应用,包括用于口服戊聚醣多硫酸钠的一有效治疗剂量,其特征在于,用于口服戊聚醣多硫酸钠的所述有效治疗剂量为1-5毫克/公斤/日。(The invention provides the use of a sodium Pentosan Polysulfate (PPS) for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects, comprising an effective therapeutic dose for oral administration of sodium pentosan polysulfate, characterized in that the effective therapeutic dose for oral administration of sodium pentosan polysulfate is 1-5 mg/kg/day.)

1. Use of sodium Pentosan Polysulfate (PPS) for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects, comprising a therapeutically effective amount for oral administration of sodium pentosan polysulfate,

the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1-5 mg/kg/day.

2. The use of sodium Pentosan Polysulfate (PPS) according to claim 1 for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects,

the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1.67-3.4 mg/kg/day.

3. The use of sodium Pentosan Polysulfate (PPS) according to claim 1 for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects,

the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1-1.67 mg/kg/day.

4. Use of sodium Pentosan Polysulfate (PPS) according to claim 3 for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects,

the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1.25-1.42 mg/kg/day.

5. Use of sodium Pentosan Polysulfate (PPS) according to claim 3 for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects,

the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1-1.25 mg/kg/day.

Technical Field

The invention relates to a pharmaceutical composition for preventing recurrent urinary tract infection

Background

Lower urinary tract infections are common infectious diseases in women due to the replacement of the normal flora in the lower urinary tract by urinary pathogenic bacteria. Lower urinary tract infections are generally treated with antibiotics, and after clinical complete recovery, approximately 20% to 30% of women who have previously experienced a lower urinary tract infection may have relapsed into urinary tract infections within one year. This condition can be defined as a recurrent urinary tract infection (rUTI). The uropathogenic bacteria are derived from extra-urinary tissue or persist in the bladder epithelium.

Current preventative measures for recurrent urinary tract infections may include antibiotic and non-antibiotic preventative measures. Chronic administration of low doses of antibiotics has been shown to be effective in preventing recurrent urinary tract infections. However, after administration of low doses of antibiotics, the said uropathogenic bacteria in the lower urinary tract may thus develop resistance. Non-antibiotic prophylaxis may include bladder perfusion (interstitial infection) hyaluronic acid (hyaluronic acid), vaginal perfusion of estrogens, and supplementation with cranberry, vitamin C, or mannose (D-mannose).

Of the non-antibiotic preventative measures listed above, the only clinically proven preventive measure of recurrent urinary tract infections that is effective is bladder perfusion with hyaluronic acid. However, bladder perfusion is a complex procedure and can only be performed by a physician.

It is therefore an object of the present invention to provide a reliable and convenient preventive measure against recurrent urinary tract infections.

Disclosure of Invention

It is an object of the present invention to provide a therapeutically effective amount of a pharmaceutical composition for the prevention of recurrent urinary tract infections.

It is an object of the present invention to provide an effective therapeutic amount of oral sodium Pentosan Polysulfate (PPS) for the prevention of recurrent urinary tract infections.

An object of the present invention is to provide a use of sodium pentosan polysulfate for the preparation of a medicament for the prevention of recurrent urinary tract infections, comprising a therapeutically effective amount of sodium pentosan polysulfate for oral administration.

The invention provides the use of sodium pentosan polysulfate for the preparation of a medicament for the prevention of recurrent urinary tract infections in human subjects, comprising a therapeutically effective dose for oral administration of sodium pentosan polysulfate, characterized in that the therapeutically effective dose for oral administration of sodium pentosan polysulfate is 1-5 mg/kg/day.

According to an embodiment of the invention, the therapeutically effective amount of the sodium pentosan polysulfate for oral administration is 1.67-3.4 mg/kg/day in a human subject.

According to an embodiment of the invention, the therapeutically effective amount of the sodium pentosan polysulfate for oral administration is 1-1.67 mg/kg/day in a human subject.

According to an embodiment of the invention, the therapeutically effective amount of the sodium pentosan polysulfate for oral administration is 1.25-1.42 mg/kg/day in a human subject.

According to an embodiment of the invention, the therapeutically effective amount of the sodium pentosan polysulfate for oral administration is 1-1.25 mg/kg/day in a human subject.

The present invention provides a method of preventing a recurrent urinary tract infection in a human subject, comprising the steps of: orally administering a therapeutically effective amount of sodium pentosan polysulfate to the human subject, wherein the therapeutically effective amount in the human subject is 1-5 mg/kg/day.

Drawings

The following drawings depict one or more embodiments of the present invention and, together with the description, serve to explain the principles of the invention. Wherein like reference numerals are used to refer to the same or similar components throughout the several views of the drawings.

FIG. 1 shows a chemical structure of sodium Pentosan Polysulfate (PPS) in accordance with one embodiment of the present invention.

FIG. 2 is a flowchart of an experiment performed on rats with oral administration of sodium pentosan polysulfate, in accordance with an embodiment of the present invention.

Fig. 3 is a bar graph comparing the bacterial counts in urine of a control group, a model group (UTI model) of urinary tract infected animals and a prophylactic group of sodium pentosan polysulfate in the oral administration of sodium pentosan polysulfate to rats, in accordance with one embodiment of the present invention.

Fig. 4 is a bar graph comparing peripheral blood leukocyte counts of each group among the control group, the urinary tract infected animal model group, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate experiment, in accordance with one embodiment of the present invention.

Fig. 5 is a bar graph comparing peripheral red blood cell counts of each group among the control group, the urinary tract infected animal model group, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate experiment, in accordance with an embodiment of the present invention.

Fig. 6 is a bar graph comparing the IL-1 β concentration levels in peripheral blood of each group among the control group, the urinary tract infected animal model group and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate experiment, in accordance with one embodiment of the present invention.

Fig. 7 is a bar graph comparing TNF-a concentration levels in peripheral blood of each group among the control group, the urinary tract infected animal model group, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate experiment, in accordance with an embodiment of the present invention.

Fig. 8A-8C are H/E staining results of bladder in the control group, the urinary tract infection animal model group and the sodium pentosan polysulfate prevention group of the rat oral administration experiment with sodium pentosan polysulfate, in accordance with an embodiment of the present invention.

FIG. 9 is a Kaplan-Meier survival curve for human subjects undergoing oral testing for sodium pentosan polysulfate, in accordance with an embodiment of the present invention.

Fig. 10 is a bar graph comparing urine bacterial counts of a control group, a normal dose group, a reduced dose group, and a model group of animals with urinary tract infections in a rat oral experiment of sodium pentosan polysulfate doses, in accordance with an embodiment of the present invention.

Fig. 11 is a bar graph comparing the peripheral blood leukocyte counts of each group among the control group, the normal dose, the reduced dose, and the model group of urinary tract-infected animals for the rat sodium pentosan polysulfate dose oral experiment, in accordance with an embodiment of the present invention.

In accordance with common practice in the art, the various features are not drawn to scale, but are drawn to emphasize features relevant to the present invention. In the drawings and the description, like reference numerals refer to like elements.

Detailed Description

The present invention now will be described more fully hereinafter with reference to the accompanying drawings, in which various embodiments of the invention are shown. The invention may be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Like reference numerals refer to like elements.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising," when used herein, specify the presence of stated features, regions, integers, steps, actions, components, and/or groups, but do not preclude the presence or addition of one or more other features, regions, integers, steps, actions, components, and/or groups thereof.

It is to be understood that the terms "and/or" and "at least one" are intended to include any and all combinations of one or more of the listed elements. Also, although the terms first, second, and third may be used herein to describe various components, assemblies, regions, sections and/or sections, these components, assemblies, regions, sections and/or sections should not be limited by these terms. These terms are only used to distinguish one component from another component, one region from another region, one component from another component, or one paragraph from another paragraph. Thus, a first element, a first region, a first component, or a first section discussed below could also be termed a second element, a second region, a second layer, or a second section without departing from the teachings of the present invention.

As used herein, the term "sodium Pentosan Polysulfate (PPS)" refers to a semisynthetic, heparinoid carbohydrate-derived macromolecule having a molecular weight of 4000-6000 daltons (Dalton). Sodium pentosan polysulfate is a white, odorless and slightly hygroscopic powder with a solubility of up to 50% in water at a pH of 6. FIG. 1 shows a chemical structure of sodium pentosan polysulfate according to one embodiment of the present invention. The chemical formula of sodium pentosan polysulfate is (C)₅H₆Na₂O₁₀S₂)_nAnd R in FIG. 1 is SO₃And (4) Na. Sodium pentosan polysulfate is also a commercially available drug:the Active Pharmaceutical Ingredient (API) of (1).

Administration of sodium pentosan polysulfate has been approved as a treatment for Interstitial Cystitis (IC), or known as Painful Bladder Syndrome (PBS), because of its chemical structure and properties similar to the glycosaminoglycan (glycoaminoglycan) layer of the bladder wall. The glycosaminoglycan layer on the bladder wall comprises hyaluronic acid (hydoronic acid), chondroitin sulfate (chondroitin sulfate), keratan sulfate (keratan sulfate), and heparin sulfate (heparin sulfate). The glycosaminoglycan layer on the bladder wall is a protective layer for epidermal cells and prevents urinary tract pathogenic bacteria from adhering to the bladder wall, thereby preventing further bacterial infection of the bladder. Sodium pentosan polysulfate is structurally similar to heparin sulfate, which is one of the glycosaminoglycans and an anticoagulant. Because of its similarity to glycosaminoglycans, sodium pentosan polysulfate acts as a protective coating for the bladder wall when excreted in urine, thus preventing urinary pathogenic germs from adhering to the bladder wall. Several common sodium pentosan polysulfate side effects include: nausea, diarrhea, indigestion, headache, erythema, dyspepsia, lower abdominal pain, abnormal liver function, dizziness, and rectal bleeding.

As used herein, the term "prevention" includes prophylactic measures of a disease in an individual prior to exposure to a pathogen or prior to the occurrence of one or more symptoms of the disease or disorder. The prophylactic methods provided herein comprise orally administering to a subject an effective amount of sodium pentosan polysulfate for a specified period of time. Suitable individuals include human individuals susceptible to the diseases described herein, or animal models prior to exposure to the causative agent of the diseases described herein.

As used herein, a "therapeutically effective dose" refers to a dose that is effective for the required period of treatment, e.g., to inhibit or prevent a bacterial infection in a particular tissue or organ, in order to achieve the desired therapeutic result. The therapeutically effective amount of sodium pentosan polysulfate may vary depending on the age, sex, weight and/or body area of the individual. The therapeutically effective amount of sodium pentosan polysulfate may also be an amount that provides a therapeutic benefit far greater than the toxic or deleterious effects of sodium pentosan polysulfate.

According to one embodiment of the present invention, there is provided a method for preventing recurrent urinary tract infections (rUTI) by orally administering the therapeutically effective amount of sodium pentosan polysulfate. In patients with recurrent urinary tract infections, each urinary tract infection causes damage to the bladder mucosa, thereby increasing the susceptibility to the next bacterial infection. Sodium pentosan polysulfate can act as a protective coating for the epidermal cells of the urinary tract when excreted into the urine. The protective coating provided by sodium pentosan polysulfate can repair the mucous membrane of the urinary tract, thus inhibiting the attachment and invasion of pathogenic bacteria of the urinary tract to the urinary tract and also promoting the repair of damaged bladder epithelium. Thus, it is possible to prevent repetitive infections caused by urinary pathogenic bacteria in the urinary tract.

According to an embodiment of the present invention, there is also provided a use of sodium pentosan polysulfate for the preparation of a medicament for the prevention of recurrent urinary tract infections, comprising a therapeutically effective amount for oral administration of sodium pentosan polysulfate.

The following experiments demonstrate the efficacy of oral administration of sodium pentosan polysulfate and the effective dose of oral administration of sodium pentosan polysulfate for preventing recurrent urinary tract infections in rats or human subjects, according to one embodiment of the present invention.

Example 1: therapeutic effect of orally administered sodium pentosan polysulfate for preventing urinary tract infection caused by pathogenic Escherichia coli (Escherichia coli) in rat

The purpose of this example was to establish a model of urinary tract infected rat, study the efficacy of oral sodium pentosan polysulfate in the bladder in the model of urinary tract infected rat, centimetre and biomarkers related to urinary tract infection, and develop new targets for preventing urinary tract infection.

This example Sprague-Dawley (SD) rats were infected with uropathogenic Escherichia coli (UPEC) to establish urologyRat animal model of tract infection. UPEC is known to be the leading cause of urinary tract infections. The virulence factors of UPEC regulate its ability to adhere and invade on bladder epithelial cells, while the flagella of UPEC enable it to escape the host's immune response. The UPEC used in this example is700336^TMAnd number J96 (serotype O4: K6), available from taiwan food industry research institute of fiscal corporation. The UPEC were stored in tryptic soy agar/broth (trypticase soy agar/broth) and centrifuged at 5000rpm for 10 minutes. The pellet of UPEC was resuspended in 10mL of sterile phosphate buffered saline (PB S). The cell density of the UPEC was adjusted to an optical density (OD600) of 1 to 1.1. The optical density is equivalent to 8 × 10⁷To 2X 10⁸Colony Forming Unit (CFU) of (1)/mL. The UPEC is added with 250 μ L of 0.9% physiological saline solution, and is formulated to form 8 × 10⁷CFU suspension, which is then used to infect rats.

30 healthy female SD rats were randomly distributed into 3 groups: a control group, a model group of animals with urinary tract infections, and a prophylactic group of sodium pentosan polysulfate, wherein each of the groups has 10 female SD rats. In fig. 3 to 7, the urinary tract infection animal model group was labeled as "UTI" and the sodium pentosan polysulfate prevention group was labeled as "UROpre".

FIG. 2 is an experimental flow chart of this example, according to an embodiment of the present invention. The control group did not take any sodium pentosan polysulfate or was infected with UPEC. The urinary tract infection animal model group and the pentosan sodium polysulfate prevention group infected UPEC via urethral catheter (trans-urethral catherization) on day 1. The sodium pentosan polysulfate prevention group is used for feeding the sodium pentosan polysulfate from-14 days to 0 days by using an 18G stainless steel feeding needle connected with a 3mL syringe in an oral gavage mode. In this example, the dose of sodium pentosan polysulfate for rats was 30 mg/kg/day. Urine samples from rats were obtained on days 1, 3, 7, 10 and 14 using a PE50 catheter by a trans-urethral catheterization; blood samples were also taken from the tail vein on days 1, 3, 7, 10 and 14 with a 23G needle attached to a 1mL syringe. Rats of this example were sacrificed on day 14. 50 u L urine samples in LB culture solution diluted 10 times, and the undiluted urine samples were each point in 3 plate LB agar plate. The LB broth and LB agar plates were incubated at 37 ℃ for 16 to 18 hours. UPEC in LB broth and UPEC in LB agar plates were quantitated after incubation. Urinary tract tissue samples were then taken from the kidney and bladder after rat death and H/E staining was used to examine the histological characteristics of the urinary tract tissue.

Fig. 3 is a bar graph comparing the urine bacterial counts of the control group, the urinary tract infection animal model group and the sodium pentosan polysulfate prevention group in accordance with one embodiment of the present invention. FIG. 3 shows that the urine bacterial counts of the control group, the urinary tract infection animal model group and the sodium pentosan polysulfate prevention group were less than 10 at day 0³CFU/mL, and the control group urine bacterial counts were maintained at 10 in all cases³CFU/mL. The urine bacterial counts of the sodium pentosan polysulfate prevention group were lower than those of the model group of urinary tract-infected animals on days 1, 3 and 7, which indicates that sodium pentosan polysulfate may inhibit the growth of UPEC in this example.

Fig. 4 is a bar graph comparing peripheral blood leukocyte counts of the control group, the urinary tract infected animal model group and the sodium pentosan polysulfate prevention group in accordance with one embodiment of the present invention. White blood cell counts in the different groups reflect the UPEC infection status of the rats: a higher white blood cell count may represent a more severe UPEC infection. The sodium pentosan polysulfate prevents the group from steadily decreasing in white blood cell count after day 3 and is lower than the white blood cell count of the model group of animals with urinary tract infections on days 1, 3 and 7. The white blood cell count in the model group of urinary tract-infected animals was the highest in the 3 groups on days 1, 3 and 7.

Fig. 5 is a bar graph comparing peripheral red blood cell counts of each group among the control group, the urinary tract infected animal model group, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate experiment, in accordance with an embodiment of the present invention. Here there was no significant difference in the red blood cell counts of the different groups of rats.

The Luminex assay herein was used to evaluate peripheral blood biomarkers associated with infection or inflammation. Fig. 6 is a bar graph comparing the IL-1 β concentration levels in Luminex assay results of the groups among the control group, the model group of urinary tract infected animals, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate assay, in accordance with an embodiment of the present invention. The level of IL-1 β concentration is associated with a current inflammatory or infectious response, and higher levels of IL-1 β concentration indicate more severe inflammation. The sodium pentosan polysulfate prevention group had significantly lower IL-1 β concentration levels on day 3 than the urinary tract infected animal model group. This represents an infection caused by UPEC that was better controlled in the sodium pentosan polysulfate prevention group than in the urinary tract infection animal model group. Fig. 7 is a bar graph comparing TNF- α concentration levels in Luminex assay results of the groups among the control group, the model group of urinary tract infected animals, and the sodium pentosan polysulfate prevention group of the rat oral sodium pentosan polysulfate assay, in accordance with an embodiment of the present invention. TNF- α is a cytokine that promotes the inflammatory response, and higher levels of TNF- α concentration indicate a more severe inflammatory response in rats. The sodium pentosan polysulfate prophylactic group exhibited lower levels of TNF- α concentration on days 3 and 14, which represented a weaker inflammatory response, compared to the urinary tract infected animal model group.

FIG. 8A shows the result of H/E staining of a bladder tissue of the control group according to an embodiment of the present invention; FIG. 8B shows the result of H/E staining of a bladder tissue of the model group of animals with urinary tract infection; fig. 8C shows the H/E staining of a bladder tissue in the sodium pentosan polysulfate prevention group. It can be observed in fig. 8C that a newly formed layer of epithelial cells has accumulated on the surface layer of the tissue. In fig. 8C, the newly formed epithelial cells accumulated on the surface of the urothelial tissue showed a protective effect of the sodium pentosan polysulfate coating also contributing to the development of a more structurally intact urothelial tissue, which is now known to be of considerable importance in combating UPEC infection.

In summary, oral administration of 30 mg/kg/day sodium pentosan polysulfate to rats for 14 days in this example was effective in reducing urinary tract infections in rats and at lower levels of inflammation-associated cytokines compared to the model group of animals with urinary tract infections.

Example 2: therapeutic effect of orally administered sodium pentosan polysulfate for preventing recurrent urinary tract infection in human subjects

This example is a multicenter, prospective, open-label, second phase randomized control trial. The purpose of this example was to investigate the efficacy of oral administration of sodium pentosan polysulfate to human subjects for the prevention of recurrent urinary tract infections. The test period of this example is 16 weeks, which should cover the majority of recurrent episodes of recurrent urinary tract infections.

The human individuals in this example were screened and recruited in the clinic. Eligible patients are 20-70 year old female patients with a history of urinary tract infections of 2 or more in the last 6 months, or 3 or more in the last 12 months, and treated with antibiotics in the first few urinary tract infections. The eligible patient must be free of lower urinary tract symptoms and have been completely recovered from the previous urinary tract infection. This example also excludes patients with the following conditions: those who had an allergic reaction to sodium pentosan polysulfate, those who had been scheduled to receive any surgical procedure within a 24-week observation period after the end of the test, those who had been subjected to any interventional clinical trial 90 days before screening, those who had been subjected to bladder perfusion with hyaluronic acid within 3 months before screening, those who were pregnant, those who had suffered from a fever above 38 ℃, those who had a serum leukocyte count above 12,000/mm³Patients who had a urinary catheter inserted into one week, patients with urinary tract abnormality (urinary tract abnormalities), patients with interstitial cystitis (interstitial cystitis), patients with renal insufficiency, patients with cancer history, and patients with immune hypofunction (immunocompromises)ed), those who have received organ transplants, those with pelvic organ prolapse (pelvic organ prolapse) at grade 2 or above grade 2, those with a post-void residual urine (greater than 100 mL), those with stress or urge urinary incontinence (stress or urinary incontinence), and those who have been positive for culture of urinary bacteria after screening. A total of 45 patients were screened for eligibility, with 27 patients included and 18 patients excluded. The screening process in this trial was defined as visit 1.

The 27 patients were randomly assigned to one pentosan sodium polysulfate group (N13) and one control group (N14). The sodium pentosan polysulfate group is orally administered daily (300 mg total daily) 3 times daily, 100 mg each time, over the first 8 weeks. The dose was then changed to 100 mg once a day 2 more times in the last 8 weeks (a total of 200 mg of sodium pentosan polysulfate was administered daily). Sodium pentosan polysulfate can be administered 1 hour before or 2 hours after a meal. Any form of prevention or treatment of urinary tract infections, including antibiotics, bladder perfusion hyaluronic acid, cranberry, non-steroidal anti-inflammatory drugs or steroid drugs, is prohibited during the trial period of the trial. One patient in the group of sodium pentosan polysulfates was dropped off immediately after randomization, and did not receive any treatment course of sodium pentosan polysulfate. The control group did not take any placebo. Patients in this group received 1 visit every 4 weeks, from week 0 (visit 2), week 4 (visit 3), week 8 (visit 4), week 12 (visit 5) to week 16 (visit 6), with visit 6 marked as the end of the trial.

At each visit, patients reported to the investigator urinary tract-related symptoms. The diagnosis of urinary tract infections is based on the symptoms reported by the patient (pain in urination, frequency, urgency, suprapubic/bladder pain (suprapubic pain), nocturia or hematuria) and urine test results (white blood cells: > 5/HPF, nitrite positive or leukocyte lipolytic enzyme (leukacyte esterase) positive). And (4) carrying out urine bacterial culture on the patient with one of the symptoms and positive urine examination result so as to confirm the urinary tract infection. Patients without recurrent urinary tract infections were subjected to additional urinary cultures at week 16 (visit 6, endpoint of the experiment). Information about Adverse Events, including their severity (according to National Cancer Institute Common health Criteria for additive Events) version 5), time of occurrence, and investigator's association of Adverse Events with drugs, were also recorded at each visit. Since sodium pentosan polysulfate has a weak anticoagulant effect, a complete blood count, liver function test, prothrombin time/international normalized ratio (PT/INR), and activated partial thromboplastin time (aPTT) were performed at visits 1, 4, and 6.

To analyze the results of this experiment, all Statistical analyses were performed using a MedCalc Statistical Software (ver 19.0.3). The Mann-Whitney U assay and the Seedton t-assay (Student's t-test) were used to compare median and mean values, respectively. Chi-square test and Fisher's exact test were used for comparison and construction of a Listing table (containment table). Survival curves were constructed using the Kaplan-Meier method. A log-rank test (log-rank test) and a Cox proportional hazard model (Cox proportional hazard model) were used to compare the survival time without recurrent urinary tract infection between each group. All assays were double-tailed (2-tailed). When the P value is < 0.05, the statistical result can be considered significant.

The primary endpoint for this trial was no recurrent urinary tract infection. In this test, patients without any of the sodium pentosan polysulfate groups had lower urinary tract symptoms or urine bacteria culture appeared positive, so there was no recurrence of urinary tract infection by any of the patients in the sodium pentosan polysulfate group. In contrast, 9 of 14 patients in the control group (approximately 64%) had recurrent urinary tract infections. Table 1 provides a demographic description of the patients and the results of the recurrence of urinary tract infections in this trial.

Table 1: patient demographics and the outcome of recurrence of urinary tract infections in this trial.

According to previous studies, common adverse reactions of oral administration of sodium pentosan polysulfate include nausea (1.4% -7.9%), diarrhea (1-14.9%), headache (1-2.9%), erythema (3%), dyspepsia (2%), lower abdominal pain (2%), liver dysfunction (1-2%), dizziness (1/%) and rectal bleeding (4%). However, no adverse reaction was observed in any of the aforementioned groups of sodium pentosan polysulfate in this test. All patients showed no abnormalities in their blood counts, liver function tests and coagulation function-related tests.

FIG. 9 is a Kaplan-Meier survival curve showing the recurrence-free survival rate of urinary tract infection of 26 patients in the control group and the sodium pentosan polysulfate group according to the present test, in accordance with one embodiment of the present invention. From fig. 9, it can be seen that the recurrence-free survival rate of all urinary tract infections in the sodium pentosan polysulfate group was significantly higher than that in the control group (log-rank test p ═ 0.0004). In this test, the sodium pentosan polysulfate composition was higher than the expected clinical efficacy, which ended the test earlier and no more patients were involved. According to this test, oral administration of sodium pentosan polysulfate in human subjects has been shown to have excellent protective effects against repeated urinary tract infections.

Example 3: therapeutic effect of orally taking low-dose sodium pentosan polysulfate for preventing urinary tract infection of rat individual

The purpose of this example is to study the efficacy of orally administering a reduced dose of sodium pentosan polysulfate to rats in preventing urinary tract infections in a rat urinary tract infected animal model. SD rats were randomly grouped into a control group (N ═ 7), a model group of animals with urinary tract infections (N ═ 10), a normal dose sodium pentosan polysulfate group (N ═ 10), and a reduced dose sodium pentosan polysulfate group (N ═ 5). The urinary tract infected animal model group, the normal dose of the sodium pentosan polysulfate group, and the reduced dose of the sodium pentosan polysulfate group were infected with UPEC on day 1. The method and the perfusion amount of UPEC for infecting rats in this experiment were the same as those in example 1. The normal dose of sodium pentosan polysulfate group is orally administered with 30 mg/kg/day of sodium pentosan polysulfate daily from-14 days to 0 days, the reduced dose of sodium pentosan polysulfate group is orally administered with 8.5 mg/kg/day of sodium pentosan polysulfate daily from-14 days to 0 days, and the urinary tract infection animal model group is not administered with sodium pentosan polysulfate. The control group was not infected with UPEC nor was given sodium pentosan polysulfate. Urine and blood samples were taken from rats on days 0, 3 and 7 using the methods and tools described in example 1. The urine samples obtained were further diluted by 10, 102, 103, 104 and 105 times, and the diluted urine samples were further incubated on BSA plates at 37 ℃ for 12 hours. After incubation, UPEC colonies were counted on BSA plates. In fig. 10 and 11, the urinary tract infection animal model group is labeled as "UTI", the normal dose sodium pentosan polysulfate group is labeled as "UROpre" and the reduced dose sodium pentosan polysulfate group is labeled as "low UROpre".

Fig. 10 is a bar graph comparing urine bacterial counts of the control group, the normal dose sodium pentosan polysulfate group, the reduced dose sodium pentosan polysulfate group, and the model group of urinary tract-infected animals, in accordance with an embodiment of the present invention. The urine bacterial counts in the low UROpre group and UROpre group were similar, with urine bacterial counts in this group being lower than those in the model group of UROpre-infected animals at day 3.

Fig. 11 is a bar graph comparing peripheral blood leukocyte counts of the control group, the normal dose sodium pentosan polysulfate group, the reduced dose sodium pentosan polysulfate group, and the model group of urinary tract-infected animals, in accordance with an embodiment of the present invention. The white blood cell count reflects the current urinary tract infection, and the percentage white blood cell count can be a relative indicator of comparing the white blood cell count before and after infection. The white blood cell count in figure 11 was used to demonstrate the efficacy of the reduced dose of sodium pentosan polysulfate in a rat urinary tract infected animal model. In fig. 11, the white blood cell counts of the control group, the reduced dose sodium pentosan polysulfate group, and the urinary tract infected animal model group were set to 100% at day 0. At day 1, the percent white blood cell count in the above 3 groups increased, reflecting that more white blood cells entered the peripheral blood after infection. At day 3, the white blood cell count of the low urophore group was approximately 120%, while the percentage white blood cell count of the model group of animals with urinary tract infections was approximately 130%. This demonstrates that UPEC infection in the low UROpre group has been effectively controlled at day 3. At day 7, the percentage white blood cell count of the lowurograph group was approximately 60%, which was significantly lower than that of the model group of animals with urinary tract infection.

In summary, an oral dose of 8.5 mg/kg/day of sodium pentosan polysulfate used in this example, which is less than 1/3 of the dose of sodium pentosan polysulfate used in example 1, was found to be effective in a rat urinary tract infected animal model. The therapeutic effect of the 8.5 mg/kg/day dose of sodium pentosan polysulfate is similar to the therapeutic effect of the 30 mg/kg/day dose of sodium pentosan polysulfate.

The foregoing examples have demonstrated that a therapeutically effective dose of sodium pentosan polysulfate can be administered orally in rat and human subjects, in accordance with various embodiments of the present invention. To control the infection of pathogenic E.coli in the urinary tract, rats must be administered 14 days of sodium pentosan polysulfate prior to infection.

Based on body weight and body surface area, a therapeutically effective dose of the sodium pentosan polysulfate in rats can be converted to an equivalent dose for human subjects. According to the publications published by Nair, a.b. and Jacob s. (Nair, a.b., & Jacob, S. (2016.) a simple practical guide for dose conversion between human animals and human. journal of basic and clinical medicine, 7(2), 27.), the therapeutically effective dose of sodium pentosan polysulfate for human subjects can be converted to a dose for rats divided by a conversion constant of 0.162, or a dose for rats can be converted to a dose for human subjects multiplied by 0.162. See table 2 for a more detailed dose conversion table between human subjects and rats.

Table 2: dose conversion between human and rat

According to table 2 and one or more embodiments of the present invention, a therapeutically effective amount of sodium pentosan polysulfate, even less than 5 mg/kg/day, exhibits sufficient efficacy in preventing repetitive urinary tract infections in human subjects. In example 2, sodium pentosan polysulfate doses for human subjects of less than 5 mg/kg/day, or a total dose of 300 mg (reference body weight of a human female subject of 60 kg) have been shown to be efficacious. Administration of a reduced dose of sodium pentosan polysulfate of about 3.34 mg/kg/day, or a total dose of 200 mg per day, for 8 weeks after the trial has also proved to be efficacious. Further, example 3 has demonstrated that rats are given a lower dose of sodium pentosan polysulfate for 14 days: 8.5 mg/kg/day, has a therapeutic effect similar to the dose of 30 mg/kg/day of sodium pentosan polysulfate in preventing urinary tract infections. According to table 2, a dose of 8.5 mg/kg/day for rats corresponds to a dose of 1.38 mg/kg/day for human subjects, whereas a dose of 30 mg/kg/day for rats corresponds to a dose of 4.86 mg/kg/day for human subjects. This means that a sodium pentosan polysulfate dose of 1.38 mg/kg/day for a human subject should also have efficacy in preventing recurrent urinary tract infections, and minimize adverse effects of sodium pentosan polysulfate.

A reduced dose of sodium pentosan polysulfate represents greater clinical compliance, since it represents fewer pills to be taken by the patient than a normal dose, and enhances the patient's willingness to take sodium pentosan polysulfate. Thus, a sustained and long-term dosing regimen for the prevention of recurrent urinary tract infections would be feasible for patients with a history of urinary tract infections.

The above description is only an embodiment of the present invention and is not intended to limit the scope of the present invention. Variations and modifications will fall within the scope of the invention as derived from the description and claims of this invention. Moreover, not all of the benefits and features disclosed herein need be achieved by each embodiment and the scope of the claims. Further, the abstract and the title herein are provided for patent file retrieval only and are not intended to limit the scope of the present invention in any way.