CN116549463A - Application of compound sulfamethoxazole tablet in preparation of saphenous-attack-resistant Pythium insidiosum medicines - Google Patents

Abstract

The application belongs to the technical field of medical treatment, and particularly relates to application of a compound sulfamethoxazole tablet in preparation of a saphenous-attack-resistant Pythium drug. The application discloses application of sulfamethoxazole in preparation of anti-Pythium insidioum medicines. In a second aspect the present application provides a combination of a first and a second aspect of the present application, it is characterized by comprising compound sulfamethoxazole. The application provides an economic and convenient saphenous-attack-resistant Pythium drug which is convenient for sequential treatment and easy to achieve effective blood concentration.

Description

Application of compound sulfamethoxazole tablet in preparation of saphenous-attack-resistant Pythium insidiosum medicines

Technical Field

The application belongs to the technical field of medical treatment, and particularly relates to application of a compound sulfamethoxazole tablet in preparation of a saphenous-attack-resistant Pythium drug.

Background

Saphenous saprophyticus is a major pathogenic source of infection of mammals with saprophyticus, animals and humans are major hosts, mainly in tropical and subtropical regions. There are currently few reports in China that diagnosis and treatment have great challenges. Infection usually occurs by agricultural or natural exposure to water contaminated with zoospores, pythium insidioum is a protozoa that has similar fungal characteristics in tissues. Such aquatic pathogens can cause disease when they enter the skin or mucous membranes. The treatment scheme is mainly based on individual case reports, retrospective study or drug sensitivity results based on animal models, and no unified and effective treatment scheme exists. Because the mold is similar to fungi, antifungal drugs are used or are drug sensitive, but the failure rate is high, which may be caused by the lack of target ergosterol of the antifungal drug by the pathogen or the need of higher blood concentration to exert the antibacterial effect.

The treatment effect of the patient infected by the saphenous procymidone is poor, scattered individual reports indicate that antifungal drugs (amphotericin B) and antibacterial drugs (such as macrocyclic esters) are effective, but the results are not verified on other patients, the mortality rate is high, most patients even need amputation or cornea transplantation, the recurrence rate is high, the hospitalization time of most patients is up to many months, and the corresponding sequential oral drug selection is not available after discharge, so that the recurrence possibility is increased, and great economic burden is brought to the patients and families.

Disclosure of Invention

In view of the above, the application discloses application of compound sulfamethoxazole tablets in preparation of saphenous-attack-resistant Pythium medicines, and provides an saphenous-attack-resistant Pythium medicine which is economical and convenient, is convenient for sequential treatment and is easy to achieve effective blood concentration.

The application discloses application of compound sulfamethoxazole in preparing a medicine for resisting saphenous-attack Pythium.

The compound sulfamethoxazole comprises sulfamethoxazole and trimethoprim.

In another embodiment, the anti-saphenous procymidone drug comprises a therapeutically effective amount of sulfamethoxazole and pharmaceutically acceptable pharmaceutical adjuvants.

Specifically, the saphenous-attack-resistant Pythium drug comprises: 12.5. Mu.g/mL of sulfamethoxazole and 2.5. Mu.g/mL of trimethoprim, or 25. Mu.g/mL of sulfamethoxazole and 5. Mu.g/mL of trimethoprim, or 50. Mu.g/mL of sulfamethoxazole and 10. Mu.g/mL of trimethoprim, or 100. Mu.g/mL of sulfamethoxazole and 20. Mu.g/mL of trimethoprim, or 150. Mu.g/mL of sulfamethoxazole and 30. Mu.g/mL of trimethoprim, or 200. Mu.g/mL of sulfamethoxazole and 40. Mu.g/mL of trimethoprim, or 250. Mu.g/mL of sulfamethoxazole and 50. Mu.g/mL of trimethoprim, or 300. Mu.g/mL of sulfamethoxazole and 60. Mu.g/mL of trimethoprim, or 400. Mu.g/mL of sulfamethoxazole and 80. Mu.g/mL of trimethoprim.

In another embodiment, the concentration of the sulfamethoxazole in the compound sulfamethoxazole is 12.5 mu g/mL-400 mu g/mL; the concentration of the trimethoprim is 2.5-80 mug/mL.

In another embodiment, the compound sulfamethoxazole has a half maximal effect concentration EC50 of 148.4 μg/mL.

In another embodiment, the compound sulfamethoxazole is in the form of an injection or tablet.

In another embodiment, the anti-saphenous procymidone drug is an oral formulation, an injectable formulation, or a topical formulation.

The application discloses an economic and effective scheme for treating Pythium insidiosum infection, which can be used for sequential treatment and is convenient to take. The compound sulfamethoxazole is also called neonomine, belongs to a middle-effect sulfanilamide medicine applied to the whole body, can competitively act on dihydrofolate synthase in bacteria with para-aminobenzoic acid, and prevents the synthesis of bacterial dihydrofolate, thereby inhibiting the growth and reproduction of bacteria. Sulfamethoxazole is well absorbed (about 90% or more of the amount administered can be absorbed) after oral administration, but is absorbed more slowly. The peak value of the blood concentration is reached 2-4 hours after the administration. The concentration of the free medicine in blood can reach 80-100 mug/ml after single oral administration of 2 g. The distribution volume of the compound sulfamethoxazole is about 0.15L/kg. After absorption, the drug is widely distributed in various extracellular fluids such as systemic tissues, pleural fluid, peritoneal fluid, synovial fluid, aqueous humor, saliva, sweat, urine, bile and the like, but cannot enter intracellular fluid. The application finds that the compound sulfamethoxazole has the effect of treating Pythium insidioum infection in vivo and inhibiting the growth of Pythium insidioicum in vitro.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.

FIG. 1 shows the antibacterial effect of 12.5. Mu.g/mL sulfamethoxazole and 2.5. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 2 shows the antibacterial effect of 25. Mu.g/mL sulfamethoxazole and 5. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 3 shows the antibacterial effect of 50. Mu.g/mL sulfamethoxazole and 10. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 4 shows the antibacterial effect of 100. Mu.g/mL sulfamethoxazole and 20. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 5 shows the antibacterial effect of 150. Mu.g/mL sulfamethoxazole and 30. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 6 shows the antibacterial effect of 200. Mu.g/mL sulfamethoxazole and 40. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 7 shows the antibacterial effect of 250. Mu.g/mL sulfamethoxazole and 50. Mu.g/mL trimethoprim on saphenous mould provided in the examples of the present application;

FIG. 8 is a graph showing the inhibition of saphenous mould by 300 μg/mL sulfamethoxazole and 60 μg/mL trimethoprim provided in the examples of the present application;

FIG. 9 is a graph showing the inhibition of saphenous mould by 400 μg/mL sulfamethoxazole and 80 μg/mL trimethoprim provided in the examples of the present application;

FIG. 10 shows the antibacterial effect of a negative control provided in the examples of the present application on Pythium insidiosum;

FIG. 11 is a graph showing the half-maximal effect concentration of sulfamethoxazole on saphenous mould.

Detailed Description

The application provides application of a compound sulfamethoxazole tablet in preparing a saphenous-attack-resistant Pythium drug, which is used for solving the technical defects of poor treatment effect and high recurrence rate of saphenous-attack-resistant Pythium drugs in the prior art.

The following description of the technical solutions in the embodiments of the present application will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art without undue burden from the present disclosure, are within the scope of the present disclosure.

Wherein, the raw materials or reagents used in the following examples are all commercially available or self-made.

The drugs or reagents used in the following examples are all commercially available or self-made; the compound sulfamethoxazole is marked as SMZ/TMP.

The Pythium insidiosum used in the following examples was cultured by conventional means; the saphenous pythium fungus solid culture medium is the conventional saphenous pythium fungus solid culture medium.

Example 1

The embodiment of the application provides application of sulfamethoxazole tablets in treating Pythium insidioum infection, which comprises the following specific steps:

one 56 year old male patient was diagnosed in month 12 of 2020 due to "2 months of skin infection of the right lower limb". There is a history of hepatitis B liver cirrhosis and spleen hyperactivity. His occupation is a farming, ever, who has caught his feet in swamps. The microbiological examination (including bacteria, fungi, tuberculosis and other pathogens) is negative. Vancomycin is applied for 10 days, and teicoplanin is cured for discharge after 5 days.

And 2021, the hospital was admitted again for "2 months of swelling pain of the right lower limb". The main sign is that severe non-recessed edema of the right lower limb is accompanied by black hardening nodules, and the skin temperature of the hardened region rises. Because the patient's immune index was abnormal (ANCA positive), not excluding immune vasculitis, the "40 mg/d of methylprednisolone" treatment was given for 14 days, with an initial improvement in symptoms, followed by an exacerbation of swelling pain. The "erysipelas" cannot be excluded after the multidisciplinary consultation. However, the clinical symptoms were not improved after sequential administration of penicillin, vancomycin and ceftriaxone. Because of the obvious pain and swelling of the lower limbs of patients, the patients have to be partially cut, with little liquid seepage, and the subcutaneous multiple ulcers can be seen to be in radial and net structures. And then, taking biopsied tissues of ulcer exudates, swabs and ulcer surfaces for microbiological and second generation sequencing inspection, wherein the second generation sequencing (next generation sequencing) results of all specimens show saphenous saprophyticus, the smears of the exudates and the swabs show that hyphae are separated, and then the tissue culture results of the biopsies show that the saprophyticus grows.

Histopathological examination showed inflammatory granulation tissue with eosinophil infiltration. PAS staining and Gomori Methenamine Silver (GMS) staining showed small amounts of fungal hyphae and spores, and NGS and subsequent whole gene sequencing were further verified as saphenous mould.

The treatment regimen was then changed to azithromycin, caspofungin and linezolid, and after one week azithromycin was discontinued and tigecycline was started. After 11 days of combined treatment with caspofungin, linezolid, tigecycline, etc., there was a reduction in ulcer surface leakage. However, based on thrombocytopenia caused by linezolid and dyspepsia caused by tigecycline, and economic difficulties of patients, the treatment regimen is changed to oral itraconazole, i.v. ceftazidime-tazobactam, for external use with tigecycline. Four days later the treatment was given up. Patients were asked to discharge on day 45, and after discharge, compound sulfamethoxazole (each tablet contained 0.4g sulfamethoxazole and 0.08g trimethoprim, 2 tablets/time, 3 times daily), mesenchyme, and itraconazole were taken. On day 87, ulcers were reduced. Starting on day 103, the patient only took sulfamethoxazole orally. On day 128, all skin ulcers had healed. So far, patients only take the compound sulfamethoxazole orally, no adverse reaction occurs, and the disease is stable.

It can be seen that the single administration of the compound sulfamethoxazole can treat the Pythium insidiosum infection; the combined administration of the compound sulfamethoxazole, the mermaiden and the itraconazole also has the effect of treating the saphenous pythium infection.

Example 2

The embodiment of the application provides an antibacterial test of compound sulfamethoxazole, which specifically comprises the following steps:

(1) Oomycete inoculation

And (3) taking a new transfer oomycete plate, punching holes on the position, which is about 1cm away from the plate wall, of the oomycete growth ring by using a puncher, picking up bacteria-carrying agar by using a toothpick, inverting the bacteria-carrying agar to the center position of the fresh Columbia blood agar plate, sealing the inoculated culture plate by using a sealing film, and culturing the culture plate in a culture box at 37 ℃ for 1-2 days.

(2) Pharmaceutical board

Commercially available compound sulfamethoxazole injection (purchased from Rogowski pharmaceutical) was diluted with crude salt to different drug concentrations (concentrations in Table 1). A100 ml conical flask is used for preparing a Columbia blood agar basal medium, the Columbia blood agar basal medium is sterilized at 121 ℃ for 15min under high pressure, after the Columbia blood agar basal medium is cooled to 55 ℃, diluted compound sulfanilamide medicines with different concentrations and sterile defibrinated sheep blood are added into the culture medium by using a micropipette, and the culture medium is uniformly mixed by shaking, and three plates (with the diameter of 90 mm) are poured for each concentration of each medicine.

(3) Observe and record the results

Inoculating oomycetes at the center of a medicine plate after cooling, setting a control group, sealing a sealing film, placing the sealing film in a 37 ℃ incubator, observing the growth condition of the oomycetes every 24 hours, measuring the growth diameters of oomycetes treated by different medicines by adopting a ruler crisscross method when the colony growth of the control group is about 2/3 (40-42 h), calculating the antibacterial rate of the concentration of the medicines according to the results shown in Table 1, and carrying out data processing by software by taking the antibacterial rate as an ordinate and the different concentrations of the medicines as an abscissa. The antibacterial conditions of different concentrations of the medicine on the saphenous mould are shown in figures 1-9, fig. 10 shows the antibacterial effect of negative control on saphenous mould.

As shown in Table 1, solid media of Pythium insignium containing different concentrations of the drug were prepared, the drug concentration was divided into 10 groups, 1 group was a solid medium containing 12.5. Mu.g/mL of sulfamethoxazole and 2.5. Mu.g/mL of trimethoprim, 2 group was a solid medium containing 25. Mu.g/mL of sulfamethoxazole and 5. Mu.g/mL of trimethoprim, 3 group was a solid medium containing 50. Mu.g/mL of sulfamethoxazole and 10. Mu.g/mL of trimethoprim, 4 group was a solid medium containing 100. Mu.g/mL of sulfamethoxazole and 20. Mu.g/mL of trimethoprim, 5 group was a solid medium containing 150. Mu.g/mL of sulfamethoxazole and 30. Mu.g/mL of trimethoprim, 6 group was a solid medium containing 200. Mu.g/mL of sulfamethoxazole and 40. Mu.g/mL of trimethoprim, 7 group was a solid medium containing 250. Mu.g/mL of sulfamethoxazole and 50. Mu.g/mL of trimethoprim, 8 group was a solid medium containing 100. Mu.g/mL of sulfamethoxazole and 20. Mu.g/mL of trimethoprim, and 5 group was a solid medium containing 50. Mu.g/mL of sulfamethoxazole and 10. Mu.g/mL of trimethoprim was a solid medium was prepared.

TABLE 1

Group of	SMZ/TMP(μg/mL)	Pure growth 1	Pure growth amount 2	Antibacterial ratio 1	Antibacterial rate 2
						1	12.5/2.5	7.2	7.3	7.69％	6.41％
2	25.0/5	7.1	7.0	8.97％	10.26％
						3	50.0/10	6.6	6.7	15.38％	14.10％
4	100/20	5.3	5.3	32.05％	32.05％
						5	150/30	3	3.05	61.54％	60.90％
6	200/40	2	1.95	74.36％	75.00％
						7	250/50	1.6	1.65	79.49％	78.85％
8	300/60	1.4	1.4	82.05％	82.05％
						9	400/80	1.1	1.2	85.90％	84.62％
10	Negative control	7.8	7.8	Without any means for	Without any means for

Note that: the 1 st group to the 9 th group are test groups, and the calculation formula of the bacteriostasis rate is as follows:

pure growth = colony mean diameter-cake diameter.

3. Determination of Compound sulfonamideHalf maximum effect concentration EC of Methazole on saphenous beancurd fungus ₅₀ As shown in FIG. 11, it is clear from FIG. 11 that EC of compound sulfamethoxazole ₅₀ 148.4.

This example shows that sulfamethoxazole and trimethoprim have the effect of inhibiting the growth of Pythium insidioum.

In summary, the failure rate of the current treatment of the humic acid is high, and scattered individual reports indicate that antifungal drugs (amphotericin B) and antibacterial drugs (such as macrocyclic esters) are effective, but the results are not verified on other patients. Patients have long hospitalization times or no good sequential oral drug selection after hospitalization, resulting in patients being more prone to relapse.

The embodiment of the application discovers that the compound sulfamethoxazole has the effect of treating the saphenous pythium infection, and the compound sulfamethoxazole is an economic and effective saphenous pythium resistant medicament which can be sequentially treated and is convenient to take.

The foregoing is merely a preferred embodiment of the present application and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present application and are intended to be within the scope of the present application.

Claims (5)

1. Application of compound sulfamethoxazole in preparing anti-saphenous pythium fungus medicine;

the compound sulfamethoxazole comprises sulfamethoxazole and trimethoprim.

2. The use according to claim 1, wherein the concentration of sulfamethoxazole in the compound sulfamethoxazole is 12.5 μg/mL to 400 μg/mL; the concentration of the trimethoprim is 2.5-80 mug/mL.

3. The use according to claim 1, wherein the compound sulfamethoxazole has a half maximal effector concentration EC ₅₀ 148.4. Mu.g/mL.

4. The use according to claim 1, wherein the anti-saphenous-mould medicament comprises a therapeutically effective amount of compound sulfamethoxazole and pharmaceutically acceptable pharmaceutical adjuvants.

5. The use according to claim 1, wherein the compound sulfamethoxazole is in the form of an injection or tablet.