Application of organosilicon quaternary ammonium salt in preparation of medical material for typing diagnosis of preadenitis disease

Abstract

The invention discloses an application of organosilicon quaternary ammonium salt in preparation of a medical material for typing diagnosis of prostatitis, the organosilicon quaternary ammonium salt can improve typing diagnosis of prostatitis, and a molecular-level or nano-pipeline biocompatible membrane is formed in urethra to prevent or destroy bacterial biomembranes, so that the prostatitis can be diagnosed more accurately, and the prostatitis can be diagnosed more accurately. Prostatic fluid flows out through the pipeline and is not polluted by bacteria and leukocytes, the typing diagnosis rate is increased, a basis is provided for prostatitis treatment, and a novel method for typing diagnosis of prostatitis is provided.

Description

Technical Field

[0001] The invention belongs to the technical field of prostatitis typing disease diagnosis, and relates to the application of urethra biocompatible material membrane in medical materials. Background Art

[0002] Current status and problems of prostatitis classification diagnosis at home and abroad. The classification diagnosis of prostatitis requires laboratory testing of prostatic fluid. However, since the prostatic fluid obtained under non-invasive conditions must be collected through the urethra, it may be contaminated by bacteria and leukocytes in the urethra, resulting in false positive or false negative results, leading to misdiagnosis and affecting the treatment effect. At present, the guidelines of various countries in the world use the "3 urines + prostatic fluid" Meares-Stamey four-cup method for prostatic fluid detection and classification. Before collecting prostatic fluid, 2 urines are collected, and after collecting prostatic fluid, 1 urine is collected. The collected prostatic fluid is separated and cultured separately to distinguish male urethra, bladder and prostate infections, and the types of prostatitis are divided accordingly. Although "3 urines + prostatic fluid" improves the diagnosis rate, the urethra itself will be contaminated by bacteria and leukocytes. The four-cup method cannot completely eliminate the contamination of bacteria and leukocytes. In addition, due to the complexity of the operation, the compliance of doctors in the application is extremely poor. The application rate of doctors in various countries in the world is less than 50%.

[0003] At present, there is no ideal method for the diagnosis and classification of prostatitis. Therefore, it is also an urgent need for clinical diagnosis and treatment to find a new method for the diagnosis and classification of prostatitis that is simple and accurate, provide a basis for the treatment of prostatitis, and improve diagnostic and treatment guidelines with higher clinical value. Summary of the invention

[0004] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0005] The invention provides application of organosilicon quaternary ammonium salt in preparing medical materials for typing and diagnosing prostatitis.

[0006] As a preferred embodiment of the use of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for typing and diagnosing prostatitis: the organosilicon quaternary ammonium salt includes organosilicon monoquaternary ammonium salt, organosilicon diquaternary ammonium salt and organosilicon di-long-chain diquaternary ammonium salt.

[0007] As a preferred embodiment of the application of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for diagnosing prostatitis: the structural formula of the organosilicon quaternary ammonium salt is:

[0008]

[0009] As a preferred embodiment of the application of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for diagnosing prostatitis: the structural formula of the organosilicon diquaternary ammonium salt is:

[0010]

[0011] Wherein, R1 and R2 are each selected from C 11 R3 is (EtO)3Si-, (MeO)3Si-, (EtO) 3 Si(CH2)nCO-, (EtO)3Si(CH2)3NHCO-, (MeO)3Si(CH2)3CO- or (MeO)3Si(CH2)3NHCO-; X is F, Cl, Br or I; Y1 and Y2 are the same or different and independently selected from O, NH; n is 3.

[0012] As a preferred embodiment of the use of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for diagnosing prostatitis: the structural formula of the organosilicon di-long chain diquaternary ammonium salt is:

[0013]

[0014] In the technical solution of the present invention, the organosilicon quaternary ammonium salt forms a self-assembled nanomaterial layer in vivo in the form of an aqueous solution. It is further preferred that the concentration of the organosilicon quaternary ammonium salt aqueous solution is 0.1-20 wt %.

[0015] The organosilicon quaternary ammonium salt of the present invention is used in the preparation of medical materials for diagnosing prostatitis.

[0016] The preferred embodiment described herein is that prostatitis includes acute and chronic prostatitis, and chronic prostatitis includes leukocyte and bacteria positive type, leukocyte positive and bacteria negative type, and leukocyte and bacteria negative type.

[0017] As a preferred embodiment of the use of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for diagnosing prostatitis: the organosilicon quaternary ammonium salt is administered in vivo.

[0018] As a preferred embodiment of the use of the organosilicon quaternary ammonium salt of the present invention in the preparation of medical materials for diagnosing prostatitis: the organosilicon quaternary ammonium salt is administered in a push injection manner.

[0019] In the technical solution of the present invention, the organosilicon quaternary ammonium salt forms a nano-scale or molecular-scale channel membrane after being injected into the urethra.

[0020] In the technical solution of the present invention: the organosilicon quaternary ammonium salt prevents or destroys the formation of bacterial biofilm.

[0021] In the technical solution of the present invention: the organosilicon quaternary ammonium salt isolates bacteria and leukocytes.

[0022] In the technical solution of the present invention: the prostatitis includes acute prostatitis and chronic prostatitis. Preferably: the chronic prostatitis includes leukocyte and bacteria positive type, leukocyte positive and bacteria negative type, and cell and bacteria negative type. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 for Figure 1 The images are scanning electron micrographs, in which: a. normal urethral membrane cells (group 1); b. urethral membranes forming antimicrobial agents on the surface of normal urethra (group 2); c. bacterial biofilms on the urethra (group 3); d. bacterial biofilms destroyed by nano antimicrobial films (group 4). DETAILED DESCRIPTION

[0024] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0025] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0026] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0027] Embodiment 1:

[0028]

[0029] Add KH791 (50g, 1.0eq) to a 1L single-mouth flask, add 250ml of dichloromethane to dissolve and cool to -5℃ to 5℃, then add N-methylmorpholine dichloromethane solution (47.83g, 2.5eq dissolved in 125ml dichloromethane) and chloroacetyl chloride dichloromethane solution (44.85g, 2.1eq dissolved in 125ml dichloromethane) at the same time, keep warm for 18h, spot plate detection, developing agent methanol: dichloromethane 1:10, iodine cylinder color development. After the reaction is completed, filter, wash the filtrate with saturated potassium dihydrogen phosphate 125ml three times, saturated sodium chloride 125ml three times, and dry the organic layer with anhydrous sodium sulfate. Dry under reduced pressure. The product is a yellow liquid, weighing 50g, with a yield of 63.3%.

[0030] The intermediate (50 g, 1.0 eq) was added to a 2L single-necked flask, and 1L isopropanol was added to heat and dissolve, and N,N-dimethyldodecyl tertiary amine (64 g, 2.5 eq) was slowly added dropwise, and refluxed at 80°C for 8 h. The plate was tested every 2 h, and the developing agent was PE: EA 2:1. After the reaction was completed, the reaction solution was cooled to room temperature and dried under reduced pressure to obtain 61.9 g of the target product, with a yield of 81.9%.

[0031] 1 H NMR(500MHz,Chloroform-d)δ8.15(t,J=6.1Hz,1H),4.32(s,2H),4.20(s,2H),3.81(q,J=8.0Hz,6H),3.66–3.58(m,6H),3.49(tdd,J=7.1,6.1,0.8Hz,2H) ,3.43(d,J=2.2Hz,11H),3.39(t,J=7.1Hz,2H),1.70(p,J=7.1Hz,2H),1.57(p ,J=7.1Hz,4H),1.38–1.19(m,46H),0.92–0.86(m,6H),0.68(t,J=7.1Hz,2H).

[0032] Example 2

[0033] The method for preparing a water-soluble self-assembly nanomaterial comprises adding a di-long-chain diquaternary ammonium salt of organic silicon (Example 1) into water and stirring (the mass concentration of the di-long-chain diquaternary ammonium salt of organic silicon is 1.5%) to obtain a water-soluble preparation. The particle size of the di-long-chain diquaternary ammonium salt of organic silicon is detected by a trifunctional particle system manufactured by Brookhaven Corporation (BIC) in New York, USA, and is found to be 37 nanometers.

[0034] Example 3

[0035] Animal urinary tract bacterial biomodel was established. Methods Twelve adult Beagle dogs aged 24 months were randomly divided into an experimental group (6 dogs) and a control group (6 dogs). The experimental group was instilled with 2.5 ml of standard pathogenic bacteria Escherichia coli (ATCC25922), and the control group was instilled with the same volume of sterile saline. Bacterial culture identification, drug sensitivity test and pathological observation were performed. Results Compared with the control group, the bacterial culture of the model group was significantly different (P<0.05). The bacteria were identified as the same type of Escherichia coli, with the highest resistance rate to ampicillin and the most sensitive to meropenem. Tissue formation in pathological sections, SEM image of the control group a. Normal urethral membrane cells (group 1); SEM image of the experimental group c. Bacterial biofilm on the urethra (group 3), a large number of inflammatory cells infiltrated under the microscope. Conclusion Escherichia coli can cause lower urinary tract infection in Beagle dogs, which is similar to lower urinary tract infection in humans. Multiple bladder instillations are a simple and effective experimental method to establish an animal model. It also confirms that the drug resistance of bacterial biofilm is one of the main causes of lower urinary tract infection.

[0036] Example 4

[0037] A 1.5% water-soluble preparation of organosilicon diquaternary ammonium salt forms a molecular or nanoscale channel biofilm in the urethra to collect prostate massage fluid (EPS). Methods 70 Beagle dogs were randomly divided into 7 groups, 10 in each group. A urinary tract infection model was established, and urine, EPS and prostate tissue were collected for bacterial culture and leukocyte microscopy. At the same time, urethral and prostate tissues were taken for histopathology and electron microscopy. Results are shown in Figure 1 (b and d). Conclusion The EPS collected through the urethra-formed biofilm can exclude the possibility of urethral contamination (false positive) and interference by urethral membrane (false negative), and the new method is feasible. Animal experiments showed that the EPS formed nano- or molecular-based biofilms after injection into the urethra. After the urethral membrane products were injected into the urethra of Beagle dogs, a uniform, dense, and regular physical antimicrobial film was formed. Figure 1 -b. Urethral membrane forms an antimicrobial film on the surface of the normal urethra (Group 2); Figure 1 -d. Bacterial biofilms were destroyed by nano-antimicrobial films (Group 4).

[0038] Embodiment 5:

[0039] Clinical trial of 1.5% water-soluble preparation of organosilicon long-chain diquaternary ammonium salt in the diagnosis and classification of prostatitis. 720 subjects with chronic prostatitis were included, and 240 of them were randomly selected from the control group for "three urine + EPS" test, and the remaining 480 test groups (240 cases of 1.5% water-soluble preparation of organosilicon long-chain diquaternary ammonium salt in test group I and 240 cases of normal saline in test group II) were tested. Urine and prostatic fluid specimens of all subjects were collected for microscopic examination and bacteriological examination. Results: Table 1. Conclusion: The new method in the test group can more accurately diagnose the classification of chronic prostatitis.

[0040] Table 1 Prostatitis detection rate in each group

[0041] Group Number of cases Leukocyte and bacteria positive Leukocyte positive and bacteria negative Leukocyte-negative and bacteria-negative Epidemiology 5－8％ 45％ 45％ One test group 240 7.1％ 47％ 42％ Second control group 240 56％ 32％ 37％

[0042] Table 1 Prostatitis detection rate in each group

[0043] Control group 240 24％ 52％ 37％

[0044] Note: There was no statistical difference between the first test group (1.5% water-soluble preparation of organosilicon di-long-chain diquaternary ammonium salt) and the epidemiological group (P>0.05), while the other two groups were statistically different from the epidemiological group (P<0.05 or 0.001)

[0045] Example 6 Biocompatibility Test

[0046] According to GB / T16886.5-2017 "Biological Evaluation of Medical Devices Part 5: In Vitro Cytotoxicity Test";

[0047] GB / T16886.1⒐2017 "Biological Evaluation of Medical Devices Part 10: Irritation and Skin Sensitization Test" conducted a biocompatibility test on Example 2, and the results are as follows:

[0048] Cytotoxicity test results

[0049]

[0050] Under the conditions of this experiment, the cell survival rate of the test solution was 88% and there was no potential cytotoxicity.

[0051]

[0052]

[0053] Under the conditions of this test, no erythema and edema appeared on the skin of the stimulated part of the test group animals, and according to the Magnusson and Kligman grading standard, the grade was less than 1. Therefore, under the conditions of this test, the test solution did not cause skin sensitization in guinea pigs.

[0054] Animal individual vaginal irritation observation and scoring:

[0055] Clinical observation:

[0056]

[0057] Histological evaluation:

[0058] Vaginal tissue reaction microscopy scoring table

[0059]

[0060] Under the conditions of this test, the rabbit vaginal irritation index of this sample is 0, and the reaction level is no irritation

[0061] The experimental sample prepared according to CN 103977435 B, after biocompatibility testing, had a cell survival rate of 35%, with potential cell fixation, which was higher than the sample prepared by the present invention. According to the Magnusson and Kligman grading standard, the grade was less than 1. Therefore, under the conditions of this test, the test solution did not cause skin sensitization in guinea pigs. According to GB / T16886.1⒐2017 "Biological Evaluation of Medical Devices Part 10: Irritation and Skin Sensitization Test", a vaginal mucosal irritation test was conducted, with an irritation index of 1, and there was a certain vaginal mucosal irritation test.

Claims

1. Application of organosilicon quaternary ammonium salts in the preparation of medical materials for the diagnosis and typing of prostatitis.

2. The use according to claim 1, characterized in that: The organosilicon quaternary ammonium salts include organosilicon monoquaternary ammonium salts, organosilicon diquaternary ammonium salts and organosilicon di-long chain diquaternary ammonium salts.

3. The use according to claim 1, characterized in that: The organosilicon di-long-chain diquaternary ammonium salt comprises the following structure:

4. The use according to any one of claims 1 to 3, characterized in that: The organosilicon quaternary ammonium salt forms a self-assembled nano material layer in the body in the form of an aqueous solution; preferably, the concentration of the organosilicon quaternary ammonium salt aqueous solution is 0.1-20wt%.

5. The use according to claim 4, characterized in that: Meet the biocompatibility requirements of medical devices, including cytotoxicity testing, skin sensitization testing, and vaginal irritation testing.

6. The use according to claim 1, characterized in that: The organosilicon quaternary ammonium salt aqueous solution is administered in vivo in the form of push injection.

7. The use according to claim 6, characterized in that: After being injected into the urethra, it forms a nanoscale or molecular-scale channel membrane.

8. The use according to claim 6, characterized in that: Prevent or destroy bacterial biofilm formation.

9. The use according to claim 6, characterized in that: The isolation included bacteria and leukocytes.

10. The use according to claim 1, characterized in that: The prostatitis includes acute prostatitis and chronic prostatitis. Preferably, the chronic prostatitis includes leukocyte and bacteria positive type, leukocyte positive and bacteria negative type, and cell and bacteria negative type.

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