Prophylactic and therapeutic agent for urinary tract infection comprising 1-deoxymannose

Abstract

The prophylactic and therapeutic agent for urinary tract infection is characterized by comprising 1-deoxymannose.

Description

Technical Field

[0001] The present invention relates to a preventive and therapeutic agent for urinary tract infection comprising 1-deoxymannose. Background Art

[0002] Urinary tract infection is a disease caused by the presence of pathogenic microorganisms in the urinary tract, such as the renal pelvis, urethra, and bladder, and is divided into cystitis and pyelonephritis according to the site of infection. In many cases, urinary tract infection is caused by bacterial pathogens, most of which are considered to be uropathogenic Escherichia coli (UPEC). Urinary tract infection is usually treated with antibiotics, however, this can lead to the emergence of antibiotic-resistant bacteria. Another known treatment is to take mannose in the form of supplements.

[0003] Urinary tract infection caused by UPEC occurs when the Fim H lectin of UPEC binds to the mannose residue at the end of the sugar chain protruding from the urothelial cells. Fim H lectin also binds to mannose. Therefore, when free mannose is present in urine, UPEC becomes more likely to bind to this mannose and less likely to bind to urothelial cells, thereby controlling urinary tract infection. UPEC that fails to bind to the urinary tract is excreted from the body with urine.

[0004] Mannose has been orally tested in human clinical trials, and the following results have been reported: Mannose derived from plant hemicellulose or a mixture of mannose and cranberry components is contained in the preventive and improving agent for urinary tract infection (Patent Documents 1 and 2). In addition, it is reported that mannose is effective in preventing cystitis cases (Non-Patent Document 1). At the same time, considering that only about 0.1% of orally administered mannose reaches the urinary tract (Non-Patent Document 2), the effect of mannose in preventing and treating urinary tract infection is limited.

[0005] In order to find a substance showing stronger anti-adhesion activity than mannose, many mannose derivatives have been developed (Non-patent Document 3). However, considering the fact that mannose derivatives are substances obtained by chemically modifying mannose, it is speculated that the small intestinal absorption rate of mannose derivatives orally administered to humans is even lower than that of mannose, which also makes it difficult for such mannose derivatives to reach the urinary tract. In addition, mannose derivatives have not yet been evaluated in clinical trials to determine how many of the mannose derivatives orally administered to humans reach the urinary tract and their effectiveness in preventing and treating urinary tract infections; therefore, their effects are questionable.

[0006] Furthermore, since urinary tract infections have a high recurrence rate, more effective prevention and treatment methods are needed.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: JP-A-2011-219469

[0010] Patent Document 2: JP-A-2012-171899

[0011] Non-patent literature

[0012] Non-patent literature 1: Antibiotics 2021, 10, 373.

[0013] Non-patent document 2: Luminacoids Research, Vol. 22, No. 2 (2018)

[0014] Non-patent document 3: Molecules 2018, 23, 1641

[0015] Non-patent literature 4: IUCrJ (2017) 4, 7-23 Summary of the invention

[0016] Problem that the invention aims to solve

[0017] As described above, many reports have been published on substances that bind to Fim H lectin, such as mannose and mannose derivatives. However, the effects of these substances on humans have not been evaluated or have been insufficiently evaluated. Under such circumstances, it is necessary to provide a preventive and therapeutic agent for urinary tract infections that can be safely administered to humans and can reach the urinary tract at an effective concentration to exhibit anti-adhesion activity against UPEC.

[0018] The object of the present invention is to provide an effective preventive and therapeutic agent for urinary tract infection that can be safely administered to humans. In the search for materials that meet these requirements, the present inventors discovered that 1-deoxymannose administered to humans is excreted in urine at a high rate; thus, the present invention was completed.

[0019] Solutions for solving problems

[0020] The present invention provides a preventive and therapeutic agent for urinary tract infection comprising 1-deoxymannose.

[0021] In the agent for preventing and treating urinary tract infections, 1-deoxymannose is preferably detected in the urine of a person after administration of the agent.

[0022] In the agent for preventing and treating urinary tract infections, the concentration of 1-deoxymannose in urine after administration of the agent is preferably higher than the concentration of mannose in urine after administration of mannose.

[0023] In the agent for preventing and treating urinary tract infections, the amount of urine after administration of the agent is preferably greater than the amount of urine after administration of mannose.

[0024] The preventive and therapeutic agent for urinary tract infection is preferably administered orally.

[0025] Effects of the Invention

[0026] The urinary tract infection preventive and therapeutic agent according to the present invention contains 1-deoxymannose so that it can reach the urinary tract at an effective concentration to exhibit anti-adhesion activity against UPEC, resulting in high preventive and therapeutic effects against urinary tract infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] [ Figure 1 ]: A graph showing the urinary 1-deoxymannose concentration and the urinary mannose concentration after oral administration of each sugar;

[0028] [ Figure 2 ]: A graph showing urine volume after oral administration of each sugar; and

[0029] [ Figure 3 ]: A diagram showing the utilization of sugar by Escherichia coli. DETAILED DESCRIPTION

[0030] 1-Deoxymannose, also known as 1,5-anhydro-D-mannitol, is a deoxy form of mannose in which the oxygen atom of the hydroxyl group at the 1-position is removed. 1-Deoxymannose is known to be a metabolite of 1,5-anhydro-D-fructose (1,5-AF). The chemical structural formula of 1-deoxymannose is shown below.

[0031] [Chemical formula 1]

[0032]

[0033] 1-Deoxymannose used in the present invention can be synthesized by reducing 1,5-anhydro-D-fructose (1,5-AF). Detection and quantification of 1-deoxymannose can be performed by high performance liquid chromatography (HPLC).

[0034] In the present invention, when 1-deoxymannose and mannose are administered at the same dose, the urinary 1-deoxymannose concentration after administration of 1-deoxymannose is preferably higher than the urinary mannose concentration after administration of mannose.

[0035] In addition, when 1-deoxymannose and mannose are administered at the same dose, the urine volume after administration of 1-deoxymannose is preferably larger than the urine volume after administration of mannose.

[0036] The administration route may be, for example, oral administration; direct administration into the body intravenously, subcutaneously or intraperitoneally by injection, dripping, etc.; or external application. For ease of administration to patients, oral administration is preferred.

[0037] The preventive and therapeutic agent may be in any form, such as tablets, fine powders, capsules, drops, powders, granules or injections, etc.; the form of the agent is not particularly limited. The preventive and therapeutic agent may also contain components required for the preparation, such as preparation carriers, excipients and stabilizers, etc.

[0038] In addition, the preventive and therapeutic agents of the present invention may also contain other preventive and therapeutic agents for urinary tract infections (eg, cranberry, lactic acid bacteria, polyphenols, ascorbic acid, etc.), other pharmacological components or nutritional components such as glucose, etc.

[0039] Hereinafter, the present invention will be described in further detail by way of examples. The present invention is not limited to these examples.

[0040] Example:

[0041] <Sample preparation>

[0042] The 1-deoxymannose used in the examples was synthesized as follows: a 1,5-AF aqueous solution was hydrogenated in the presence of a Raney nickel catalyst, and the resulting solution containing a reduction product of 1,5-AF was concentrated and crystallized, thereby obtaining a 1-deoxymannose sample. The sample thus obtained was confirmed to have a purity of 99% or more by HPLC. The 1-deoxymannose sample was used as a test sample.

[0043] In the comparative example, D-mannose (pure powder) manufactured by NOW Foods was used as a mannose sample. The mannose sample was confirmed to have a purity of 99% or more by HPLC. The mannose sample was used as a test sample.

[0044] <HPLC analysis conditions>

[0045] HPLC apparatus: PU2089 plus, AS2055 plus, and RI2031plus (serial numbers) manufactured by JASCO Corporation; MODEL 556 manufactured by GL Sciences Inc.

[0046] Column: MITSUBISHI MCI GEL CK08S; two-column

[0047] Oven temperature: 60℃

[0048] Eluent: Water

[0049] Detector: Differential refractometer

[0050] Flow rate: 1ml / min

[0051] Injection volume: 100 μL

[0052] Quantification of 1-deoxymannose was performed using a calibration curve prepared by using 1-deoxymannose prepared as described above. Quantification of mannose was performed using a calibration curve prepared by using premium mannose as a reagent.

[0053] <1- Safety evaluation of deoxymannose>

[0054] 1,5-AF was orally administered to humans, and then metabolites in urine were measured. As a result, about 5% of the administered 1,5-AF was recovered in urine as 1-deoxymannose. It is presumed that the orally administered 1,5-AF is reduced in vivo to form 1-deoxymannose as a product. Therefore, the 1,5-AF administration test was used as a safety evaluation test of 1-deoxymannose as a metabolite.

[0055] <Example 1>

[0056] 1 g of 1-deoxymannose was dissolved in water to prepare a 20% w / w solution as an oral administration sample. Urine was collected from six human subjects before the sample was administered. The total amount of the sample was orally administered to each subject, and the total amount of urine was collected 1, 2, 3, 4, 6, and 8 hours after administration. The urine volume of the urine collected at each time point was measured, and then filtered through a membrane filter with a pore size of 0.2 μm, and then the 1-deoxymannose concentration in the urine was determined by HPLC analysis.

[0057] <Comparative Example 1>

[0058] Mannose was orally administered in place of 1-deoxymannose in the same manner as in Example 1, and then urine was collected.

[0059] <1-deoxymannose concentration in urine>

[0060] Figure 1 The results of measuring the urinary 1-deoxymannose concentration at each time point after administration of a sample containing 1-deoxymannose in Example 1 and the urinary mannose concentration at each time point after administration of a sample containing mannose in Comparative Example 1 are shown. Each concentration in urine at each time point is a simple average of each concentration in urine of six subjects.

[0061] like Figure 1As shown, 1-deoxymannose was detected in the urine of all subjects collected from 1 to 8 hours after administration. The concentration of 1-deoxymannose was the highest at 2.12 mg / ml 2 hours after administration. In contrast, mannose was not detected from any subject. It is speculated that this is because the concentration of mannose is lower than the detection limit of HPLC, 0.1 mg / ml. In addition, the mannose content in urine was also determined by D-mannose / D-fructose / D-glucose kit (Megazyme Ltd.; detection limit: less than 10 μg / ml) using the urine supernatant obtained after urine centrifugation (10,000g, 5 minutes); however, mannose was still not detected. This proves that 1-deoxymannose reaches the urinary tract at a higher concentration than mannose, and therefore shows much higher anti-infective activity against UPEC than mannose.

[0062] The dissociation constant (K) of 1-deoxymannose for Fim H lectin D ) is reported to be 1125 nM (185 ng / ml) (Non-patent Document 4). However, the highest urinary 1-deoxymannose concentration observed 2 hours after administration was D Therefore, it was demonstrated that orally administered 1-deoxymannose exhibited a high anti-adhesion activity against Escherichia coli.

[0063] <Urine volume>

[0064] Figure 2 The results of measuring the urine volume at each time point after administration of a sample containing 1-deoxymannose in Example 1 and the urine volume at each time point after administration of a sample containing mannose in Comparative Example 1 are shown. Figure 2 In the graph, the vertical axis represents the percentage of the urine volume after administration of 1-deoxymannose in Example 1 at each time point relative to the urine volume after administration of mannose in Comparative Example 1 taken as 100%.

[0065] The results showed that the urine volume of the 1-deoxymannose administration group was greater than that of the mannose administration group at all time points. Considering that UPEC is excreted from the body through urine, the greater the urine volume, the lower the risk of UPEC infection. It is believed that the administration of 1-deoxymannose not only inhibits UPEC infection due to 1-deoxymannose excreted in urine, but also helps to effectively excrete UPEC from the body by increasing urine volume.

[0066] <Recovery rate in urine>

[0067] The urinary recovery rate of 1-deoxymannose was determined by dividing the amount of 1-deoxymannose recovered obtained from the amount of urine collected and the urinary 1-deoxymannose concentration by the dose. The urinary recovery rate is expressed by the following formula:

[0068] Recovery rate in urine [%] =

[0069] ((V 1 ×C 1 )+(V 2 ×C 2 )+(V 3 ×C 3 )+(V 4 ×C 4 )+(V 6 ×C 6 )+(V 8 ×C 8 )) / M×100

[0070] V n : Urine volume n hours after administration [ml]

[0071] C n : 1-deoxymannose concentration in urine n hours after administration [mg / ml]

[0072] M: 1-deoxymannose dosage (=1000 mg)

[0073] Up to 8 hours after administration, the urinary recovery of 1-deoxymannose was very high at 43.3 ± 9.03% when calculated as described above.

[0074] <Example 2 and Comparative Examples 2 to 3>

[0075] It is reported that the presence of glucose in the urine or urinary tract of diabetic patients etc. increases the possibility of bacterial proliferation compared to the absence of glucose in the urinary tract because such glucose serves as an energy source for bacteria. Therefore, it is expected that sugar administered for the purpose of preventing urinary tract infection will not be easily utilized by bacteria when it reaches the urinary tract.

[0076] To evaluate the utilization of 1-deoxymannose by Escherichia coli (NBRC 3301), the following experiment was performed.

[0077] Use M9 minimal medium containing 2 ml of 1% glucose (MgSO 4 and CaCl 2 Add to Difco TM M9 Low Salt (Difco TMM9 Minimal Salt). Escherichia coli (NBRC 3301) was shaken at 37°C for 17 hours to prepare a culture solution. 10 μL of the culture solution was added to 2 ml of M9 basal medium containing 1% 1-deoxymannose, 1% mannose and 1% glucose, respectively, and then cultured at 37°C with shaking. During the culture period, 100 μL of the culture solution was sampled over time and its turbidity (600 nm) was measured using a colorimetric cell with an optical path length of 1 cm. The growth of Escherichia coli was evaluated based on the increased turbidity. The measurement results are shown in Figure 3 In which Example 2, Comparative Example 2 and Comparative Example 3 correspond to a culture solution containing 1-deoxymannose, a culture solution containing mannose and a culture solution containing glucose, respectively.

[0078] The turbidity of the culture solution containing 1-deoxymannose did not increase compared to the culture solutions containing mannose and glucose, respectively. Considering that the increase in turbidity indicates an increase in the growth of E. coli, it was found that 1-deoxymannose is less likely to be utilized by E. coli (NBRC 3301) than glucose and mannose. This proves that 1-deoxymannose is a sugar suitable for a preventive and therapeutic agent for urinary tract infections.

Claims

1. A preventive and therapeutic agent for urinary tract infection, comprising 1-deoxymannose.

2. The preventive and therapeutic agent for urinary tract infection according to claim 1, wherein 1-deoxymannose is detected in urine of a human after administration of the agent.

3. The preventive and therapeutic agent for urinary tract infections according to claim 1 or 2, wherein the concentration of 1-deoxymannose in urine after administration of the agent is higher than the concentration of mannose in urine after administration of mannose.

4. The preventive and therapeutic agent for urinary tract infection according to claim 1 or 2, wherein the urine volume after administration of the agent is greater than the urine volume after administration of mannose. The preventive and therapeutic agent for urinary tract infection according to claim 1 or 2, which is administered orally.

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