CN115212192B - Application of diphenyl ketone derivative in preparation of kidney fibrosis resisting medicine - Google Patents

Abstract

The invention provides application of a benzophenone derivative in preparing medicines for resisting kidney fibrosis, wherein the benzophenone derivative is any one of 4,5,2' -trihydroxy-2, 5' -dibromobenzophenone, 3, 4-dihydroxyl-4 ' -tertiary butyl benzophenone, 4, 5-dihydroxyl-2-bromo-4 ' -tertiary butyl benzophenone and 2,3, 4-trihydroxy-6-chloro-4 ' -tertiary butyl benzophenone. Pharmacological tests prove that the benzophenone derivative can effectively reduce the kidney fibrosis degree of UUO rats and reduce abnormal deposition of ECM, and the levels of the main treatment indexes Scr and BUN are superior to those of the first-line clinical drug losartan potassium.

Description

Application of diphenyl ketone derivative in preparation of kidney fibrosis resisting medicine

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, relates to a novel pharmaceutical application of a compound, in particular to a benzophenone compound and derivatives thereof, including pharmaceutically acceptable salts thereof, and application of the benzophenone compound in preparation of medicines for resisting kidney fibrosis.

Background

Fibrosis is defined as excessive deposition of extracellular matrix proteins (Extracellular Matrix, ECM), scarring and thickening of tissue, essentially an excessive wound healing response, which can lead to dysfunction, morbidity and mortality in the patient's kidney, lung, liver, etc. vital organs. Therefore, inhibition of excessive deposition of ECM is of great importance in improving fibrosis of body organs.

Chronic kidney disease (Chronic Kidney Disease, CKD) is a long-term disease of abnormal renal tissue structure and function affecting about 10% of adults worldwide, with about 120 tens of thousands of patients dying from CKD each year. Diabetes and hypertension are two major causes of CKD, the kidneys have the ability to regenerate after an acute injury, but in the case of chronic injury regeneration and recovery are much more difficult. This process is therefore often irreversible and can progress to end stage renal disease, which requires dialysis or kidney transplantation.

The pathological hallmark of CKD is glomerulosclerosis and tubular interstitial fibrosis caused by excessive deposition of ECM, and the presence of fibrosis in CKD is closely related to future manifestations of renal failure and thus to poor long-term prognosis.

At present, no medicine for effectively treating or preventing kidney fibrosis exists, and clinical treatment mainly comprises controlling primary diseases, eliminating relevant dangerous factors (anti-infection and the like), controlling symptoms and the like, and only delaying the progress of diseases. Therefore, in order to reverse or eliminate kidney fibrosis, development of new therapeutic drugs is imperative, and in particular, discovery of drugs that reduce ECM deposition is a problem that is currently in need of solution.

ECM mainly includes collagen and non-collagen. Collagen 1 (collagen 1, COL 1) is the most abundant protein in the fibrotic matrix, fibronectin (FN) is a non-collagenous glycoprotein, and the degree of renal fibrosis is aggravated, so too is the expression of COL1 and FN. Therefore, COL1 and FN are used as indicators for evaluating the degree of kidney fibrosis.

CN 101602657a discloses a halogenated hydroxy aromatic ketone compound, its preparation method and application, the compound is that two phenyl groups of diphenyl ketone are respectively connected with one or more hydroxy and halogen substituent groups, typical compounds include 4-chloro-3 ',4' -dihydroxydiphenyl ketone, 2, 3-dibromo-4, 5-dihydroxydiphenyl ketone, 2, 6-dibromo-3, 3',4', 5-pentahydroxy diphenyl ketone, etc. The halogenated hydroxy aryl ketone compounds and non-toxic salts, esters and ethers thereof have antibacterial, antitumor, antioxidant, vascular endothelial cell injury resisting and vascular smooth muscle cell proliferation inhibiting activities.

Further, CN 102423308A discloses the application of two bromophenol compounds, 2, 3-dibromo-4, 5-dihydroxybenzophenone and 2,3 '-dibromo-4, 5,6' -trihydroxybenzophenone, and pharmaceutically acceptable salts thereof, in preparing medicaments for protecting myocardial ischemia reperfusion injury, and the protection function of the medicaments on myocardial ischemia reperfusion injury is clarified.

While CN 103156829A discloses the application of polyhydroxy bromo-benzophenone compound 2,4',5' -trihydroxy-5, 2' -dibromobenzophenone and derivatives thereof in the treatment and prevention of atherosclerosis, and proves that the compound has wider prospect for clinical medication by reducing the expression of TG and LDL-C in serum, inhibiting the expression of inflammatory factors IL-6, TNF-alpha and adhesion molecule sICAM-1 in serum, inhibiting vascular endothelial injury and protecting vascular endothelial, thereby inhibiting the occurrence and development of AS lesions.

CN 105193795a further discloses the application of two halophenol compounds, namely 2,4',5' -trihydroxy-5, 2 '-dibromobenzophenone and 2', 5-dibromo-2- (2-phenylimidazol-1-yl) methyl-4 ',5' -dihydroxybenzophenone in the aspect of angiogenesis promotion, which can remarkably increase the length of internode blood vessels of the damaged zebra fish, has a protective effect on the zebra fish with vascular damage, and has potential application prospects in the aspects of preventing and treating ischemic heart disease, stroke, muscle atrophy, distal limb necrosis and the like.

Further, CN 105816446a and CN 107007610a disclose the application of two halophenol compounds 4,5,2 '-trihydroxy-2, 5' -dibromobenzophenone and 4,5,2 '-tris (4-morpholinyloxy) -2,5' -dichlorobenzophenone in preparing anti-type ii diabetic nephropathy drugs and preparing drugs for treating acute pyelonephritis and chronic nephritis, respectively, showing the important application prospects in treating diabetic nephropathy and treating acute pyelonephritis and chronic nephritis.

Although the research shows that 4,5,2 '-trihydroxy-2, 5' -dibromobenzophenone and the like have good treatment effects on diabetic nephropathy rats, acute pyelonephritis rats and the like, the mechanism is that the compound has strong anti-inflammatory effect, and can obviously inhibit inflammatory reactions of kidney tissue cells and vascular endothelial cells. Therefore, given the apparent difference between the pathogenesis of renal fibrosis and that of diabetic nephropathy and acute pyelonephritis, the anti-renal fibrosis effect cannot be deduced from the fact that these benzophenone compounds can be used for treating diabetic nephropathy and acute pyelonephritis. In fact, the current clinically relevant drugs for the treatment of the above-mentioned kidney diseases are of little effect in reversing or slowing down kidney fibrosis.

In addition, CN 111393277a discloses 4-butyl polyhydroxy benzophenone compound derivatives and application thereof, which are used for preventing and treating oxidative stress injury, reducing blood lipid and protecting liver. Most of 4-butyl polyhydroxy diphenyl ketone derivatives provided by the preparation method have the protection rate of more than 90% on injured cells, have remarkable liver protection effect, have an inhibiting effect on adipogenesis and have a remarkable improvement effect on liver injury caused by NAFLD.

However, there is no report about the pharmacological actions of the 4-butyl polyhydroxy diphenyl ketone derivatives on resisting kidney fibrosis at home and abroad.

Disclosure of Invention

The invention aims to provide a new application of a benzophenone derivative in medicines for resisting kidney fibrosis, so as to solve the problem that medicines for resisting kidney fibrosis are lacking in the prior art.

The benzophenone derivative and the pharmaceutically acceptable salt thereof obtained by screening can be used for resisting kidney fibrosis and the application of the benzophenone derivative in preparing medicines for resisting kidney fibrosis is provided.

Further, the benzophenone derivative is specifically any one of 4,5,2' -trihydroxy-2, 5' -dibromobenzophenone, 3, 4-dihydroxyl-4 ' -tert-butylbenzophenone, 4, 5-dihydroxyl-2-bromo-4 ' -tert-butylbenzophenone and 2,3, 4-trihydroxy-6-chloro-4 ' -tert-butylbenzophenone.

More specifically, among the above benzophenone derivatives:

4,5,2 '-trihydroxy-2, 5' -dibromobenzophenone has the structural formula:

abbreviated as HP1.

The structural formula of the 3, 4-dihydroxy-4' -tert-butyl benzophenone is as follows:

abbreviated as HP2.

The structural formula of the 4, 5-dihydroxyl-2-bromo-4' -tert-butylbenzophenone is as follows:

abbreviated as HP3.

The structural formula of the 2,3, 4-trihydroxy-6-chloro-4' -tert-butyl benzophenone is as follows:

abbreviated as HP4.

The four benzophenone derivatives listed above according to the invention are all compounds reported in the literature and the preparation methods thereof are well known to the person skilled in the art. For example, a specific preparation of 4,5,2 '-trihydroxy-2, 5' -dibromobenzophenone (HP 1) and its sodium salt is given in CN 103156829A.

Further, the invention also provides application of the benzophenone derivative in preparing medicines for reducing extracellular matrix deposition.

In the kidney fibrosis resistant medicine prepared by the benzophenone derivative, the medicine active ingredient can be the benzophenone derivative or pharmaceutically acceptable salt thereof.

Further, the pharmaceutically acceptable salts thereof are specifically sodium, potassium or ammonium salts.

As with conventional medicines, the medicine provided by the invention is used for resisting kidney fibrosis and comprises necessary auxiliary materials besides the medicinal active ingredients. The auxiliary materials may include, but are not limited to, at least one of diluents, excipients, fillers, binders, humectants, absorption enhancers, surfactants, lubricants, stabilizers, flavoring agents, sweeteners, pigments, and the like, which are conventional in the pharmaceutical arts.

Furthermore, the medicament of the invention can be prepared into any pharmaceutically acceptable dosage form, and the dosage form comprises any one of tablets, capsules, granules, pills, dripping pills, powder, oral liquid, sustained release preparation or controlled release preparation.

Pharmacological tests on the four benzophenone derivatives provided by the invention prove that the four benzophenone derivatives are used as medicines for resisting kidney fibrosis, can obviously reduce serum creatinine (Scr) and serum urea nitrogen (BUN) levels of UUO rats, and improve kidney functions.

Specifically, the medicine disclosed by the invention can obviously reduce the collagen fiber area of UFO rat kidney and reduce the expression quantity of COL1 and FN in ECM protein, thereby inhibiting ECM deposition of kidney tissues and playing an anti-kidney fibrosis role.

The test result also shows that the effect of the medicine of the invention on resisting kidney fibrosis is better than losartan potassium.

Drawings

Fig. 1 is the Masson staining results of benzophenone derivatives to reduce fibrosis of kidney tissue of UUO rats.

FIG. 2 is a graph showing the results of benzophenone derivatives in reducing COL1 and FN protein expression in the ECM deposition in kidney tissue of UFO rats.

Detailed Description

The following describes in further detail the embodiments of the present invention with reference to the drawings and examples. The following examples are presented only to more clearly illustrate the technical aspects of the present invention so that those skilled in the art can better understand and utilize the present invention without limiting the scope of the present invention.

The production process, the experimental method or the detection method related to the embodiment of the invention are all conventional methods in the prior art unless otherwise specified, and the names and/or the abbreviations thereof are all conventional names in the field, so that the related application fields are very clear and definite, and a person skilled in the art can understand the conventional process steps according to the names and apply corresponding equipment to implement according to conventional conditions or conditions suggested by manufacturers.

The various instruments, equipment, materials or reagents used in the examples of the present invention are not particularly limited in source, and may be conventional products commercially available through regular commercial routes or may be prepared according to conventional methods well known to those skilled in the art.

Example 1: benzophenone derivatives were studied for their resistance to rat kidney fibrosis.

Male SD rats weighing 180-220 g, animal origin: s Bei Fu (beijing) biotechnology limited, license number: SCXK- (jing) 2019-0010, pass number: 110324220102652614.

animals and related treatments used in this experiment met the requirements of animal welfare, and prior to the development of the experiment, the institutional animal welfare committee had been subjected to ethical review.

Losartan potassium tablet, zhejiang Huahai pharmaceutical industry Co., ltd; rat creatinine (Cr) determination kit (sarcosine oxidase method, microplate method), urea nitrogen (BUN) test kit (urease method), nanjing built bioengineering institute.

72 SD rats were selected and 8 rats were randomly selected as Sham surgery group (Sham), and the remaining rats were operated on unilateral ureteral obstruction (UUIO).

After the SD rat is anesthetized by intraperitoneal injection of 3% pentobarbital sodium solution (dosage of 0.15mL/100 g), the SD rat is fixed in a supine position, the middle position of the left abdomen is opened, and a ureter is searched downwards along the kidney gate; the left ureter is dissociated, permanent ligation is respectively carried out at the opening of the renal pelvis and the 1/3 position on the ureter by using a 4-0 surgical suture, and then the left ureter is cut off from the middle of the two ligation points; suturing layer by layer, closing abdomen, and preparing into a single-sided ureteral obstruction (UUO) rat model.

Sham operated SD rats were free of ureters after opening the abdomen, but were not ligated and were directly closed.

The UFO model rats successfully prepared were randomly divided into 8 groups of model group (UFO), positive drug group (UFO+losartan potassium, 10 mg/kg/d), HP1 low dose group (22.5 mg/kg/d), HP1 medium dose group (45 mg/kg/d), HP1 high dose group (90 mg/kg/d), HP2 high dose group (90 mg/kg/d), HP3 high dose group (90 mg/kg/d) and HP4 high dose group (90 mg/kg/d), 8 animals each.

Each drug group was given its own concentration of drug for 21 days by continuous intragastric administration. The sham group and the model group were given equal doses of physiological saline.

After administration by molding, the patients are fasted overnight on day 20 and anesthetized in abdominal cavity on day 21, and relevant indexes of abdominal aortic blood detection are extracted. The kidney was washed with physiological saline, and kidney tissue (including glomeruli and tubules) of the same portion of the left kidney was longitudinally cut and immersed in 4% paraformaldehyde for pathological morphological analysis.

Analysis of data using SPSS Statistics 20 software, all values were inThe comparison between groups is shown to be a one-way analysis of variance (ANOVA) based on the results of the variance test, e.g., the variance test is a LSD test, e.g., the variance test is a Dunnett's T3 testPA difference of < 0.05 is statistically significant.

Masson staining is one of the main methods of pathological staining for judging renal fibrosis. The Masson staining results of fig. 1 show that blue collagen fiber area of the tubular stroma of the UUO rats is significantly increased compared to Sham group, while HP1 can dose-dependently decrease collagen fiber area compared to UUO group, indicating that HP1 can alleviate kidney fibrosis of UUO rats. It can also be seen from fig. 1 that the effect of high doses of HP1, HP2, HP3 and HP4 on reducing kidney fibrosis is superior to that of losartan potassium which is the current first-line clinical medicament of CKD.

Collagen 1 (COL 1) and Fibronectin (FN) are the main indicators for determining ECM deposition changes. By adopting immunohistochemistry to detect the expression levels of COL1 and FN in pathological sections of kidney tissues, the results in FIG. 2 show that compared with the Sham group, the expression levels of COL1 and FN in the UUO group are obviously increased, while the expression levels of COL1 and FN in the HP1 treatment group are obviously lower than those in the UUO group, which indicates that HP1 can effectively reduce ECM deposition in kidney tissues. It can also be seen from fig. 2 that the effect of high doses of HP1, HP2, HP3 and HP4 on reducing COL1 and FN protein expression was superior to that of the positive drug losartan potassium.

Serum creatinine (Scr) and serum urea nitrogen (BUN) are important indicators reflecting kidney function. The biochemical blood detection results in table 1 show that, compared with Sham group, the Scr and BUN contents in UUO group are significantly increased, HP1 can significantly reduce the levels of kidney injury markers Scr and BUN, and significantly improve kidney function, while the difference between the level of losartan potassium reduced Scr and UUO group has no statistical significance.

Meanwhile, the effect of improving kidney function of the high-dose HP1, HP2, HP3 and HP4 groups is obviously better than that of losartan potassium groups.

Note that: compared to the Sham group, the number,P< 0.01; in comparison with the group of UUOs, ^# ，P＜0.05， ^## ，P< 0.01; compared with the UFO+losartan potassium group, ^Δ ，P＜0.05。

the above embodiments of the invention are not intended to be exhaustive or to limit the invention to the precise forms disclosed. Various changes, modifications, substitutions and alterations may be made by those skilled in the art without departing from the principles and spirit of the invention, and it is intended that the invention encompass all such changes, modifications and alterations as fall within the scope of the invention.

Claims (4)

1. The application of a benzophenone derivative in preparing medicines for resisting kidney fibrosis is that the benzophenone derivative is any one of 4,5,2' -trihydroxy-2, 5' -dibromobenzophenone, 3, 4-dihydroxyl-4 ' -tert-butylbenzophenone, 4, 5-dihydroxyl-2-bromo-4 ' -tert-butylbenzophenone and 2,3, 4-trihydroxy-6-chloro-4 ' -tert-butylbenzophenone.

2. The use according to claim 1, wherein the medicament further comprises a pharmaceutically acceptable salt thereof.

3. Use according to claim 2, characterized in that the pharmaceutically acceptable salt is a sodium, potassium or ammonium salt.

4. The use according to claim 1, characterized in that the medicament further comprises necessary auxiliary materials.