Use of 2,6-bis(2-(trifluoromethyl)benzylidene)cyclohexanone in the manufacture of a medicament

Abstract

The application discloses application of (2E, 6E)-2, 6-bis (2-(trifluoromethyl) benzyl) cyclohexanone in preparation of a medicine, and the medicine is used for treating or preventing chronic kidney disease, preferably high blood fat and high blood pressure induced kidney disease, but is not limited to the two inducers, and specifically, the medicine can inhibit or improve pathological damage, oxidative stress and autophagy of kidney tissues.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the use of a compound of formula (I) (code name C66) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of chronic kidney disease.

[0002] Background Technology

[0003] Chronic kidney disease (CHD) is one of the leading causes of death and morbidity among people with hypertension and hyperlipidemia. There are many contributing factors to CHD, including chemical-induced, congenital, and comorbid causes. CHD is characterized by progressive glomerular sclerosis, oxidative stress in renal tissue, and autophagy, accompanied by proteinuria and a decline in glomerular filtration rate (GFR), ultimately leading to end-stage renal failure.

[0004] Based on years of work, the inventors have discovered that compound (C66) of formula (I) can effectively treat chronic kidney disease induced by various factors, especially kidney disease in patients with long-term hyperlipidemia or high Ang II, and can inhibit kidney damage induced by hyperlipidemia or high Ang II. Summary of the Invention

[0005] The object of this invention is to provide new uses for (2E,6E)-2,6-bis(2-(trifluoromethyl)benzylmethyl)cyclohexanone (a compound of formula (I), coded C66).

[0006] Specifically, the present invention provides the use of a compound of formula (I) (C66) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment or prevention of chronic kidney disease.

[0007]

[0008] The preferred use of the present invention is in the preparation of medicaments for the treatment or prevention of chronic kidney disease.

[0009] The present invention also provides the use of a compound of formula (I) (C66) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving chronic kidney disease.

[0010]

[0011] The preferred use of the present invention is in the preparation of a medicament for improving chronic kidney disease.

[0012] Kidney disease is caused by a variety of factors, including urinary tract obstruction, hyperlipidemia, hypertension, immune system disorders, and drug-induced kidney disease. The symptoms and treatments differ depending on the cause. Preferably, the kidney disease addressed in this invention is primarily caused by, but not limited to, hyperlipidemia or hypertension.

[0013] Preferably, in the use of this invention, the main symptoms of kidney disease are glomerular sclerosis, oxidative stress in kidney tissue, and autophagy.

[0014] Preferably, in the use of the present invention, the compound (C66) of formula (I) or a pharmaceutically acceptable salt thereof does not statistically significantly reduce blood lipids or blood pressure.

[0015] The medicament used in this invention contains an effective dose of a compound of formula (I) (C66). The effective dose can be the content in a unit dose form (e.g., a tablet, injection, pill, or injection) or a unit dose for the patient requiring treatment / prevention (e.g., a dose per unit body weight). The medicament manufacturer can easily convert the dose per unit body weight of the patient group requiring treatment / prevention into the content in a unit dose form of the medicament using the average weight of the patient group. For example, the average weight of an adult patient might be 60 kg; therefore, the content in a unit dose form for adults can be obtained by multiplying the average weight by the adult's dose per unit body weight. In this invention, the patient can be a mammal, such as a human, rabbit, dog, or mouse. Based on the equivalent dose conversion relationship between experimental animals and humans known to those skilled in the art (usually referring to the guidance of drug regulatory agencies such as the FDA and SFDA, and also to "Huang Jihan et al. Equivalent dose conversion between animals and between animals and humans in pharmacological experiments. Chinese Journal of Clinical Pharmacology and Therapeutics, 2004, 9(9):1069-1072"), the human dose per unit body weight can be derived from the dose of experimental animals. For example, for the commonly used experimental animal, the mouse, according to the above literature, has a conversion relationship of approximately 12:1 with adults; for the commonly used experimental animal, the rat, according to the above literature, has a conversion relationship of approximately 6:1 with adults. In this invention, the effective dose (in terms of content) can be 10ug-1g, preferably 0.1mg-500mg, more preferably 1mg-100mg.

[0016] The pharmaceutical compositions used in this invention typically also contain pharmaceutically acceptable carriers. As used herein, pharmaceutically acceptable carriers refer to non-toxic fillers, stabilizers, diluents, adjuvants, or other pharmaceutical excipients. Examples include diluents and excipients such as water and physiological saline; fillers such as starch and sucrose; binders such as cellulose derivatives, alginate, gelatin, and / or polyvinylpyrrolidone; humectants such as glycerin; disintegrants such as agar, calcium carbonate, and / or sodium bicarbonate; absorption enhancers such as quaternary ammonium compounds; surfactants such as hexadecyl alcohol; adsorbent carriers such as kaolin and / or soap clay; and lubricants such as talc, calcium / magnesium stearate, and polyethylene glycol. Furthermore, the pharmaceutical compositions of this invention may further contain other excipients, such as flavoring agents and sweeteners. According to known techniques in the art, pharmaceutical compositions can be formulated into various dosage forms according to the therapeutic purpose and route of administration. Preferably, the composition is in a unit dosage form, such as a lyophilized form, tablet, capsule, powder, emulsion, injection, or spray. More preferably, the pharmaceutical composition is in an injectable dosage form (e.g., lyophilized powder for injection) or an oral dosage form (e.g., tablet, capsule). The drug can be administered via conventional routes, particularly enterically, such as orally, in tablet or capsule form, or parenterally, such as in injectable solutions or suspensions, topically, such as in lotions or gels, or in nasal or diagnostic forms.

[0017] For ease of understanding, this invention references publicly available documents, the entire contents of which are incorporated herein by reference. The invention will now be described in detail with reference to specific embodiments and accompanying drawings. It should be particularly noted that these descriptions are merely partial exemplary descriptions and do not constitute a limitation on the scope of the invention. Many variations and modifications of the invention will be apparent to those skilled in the art based on the discussion in this specification. Attached image description:

[0018] Figure 1 The effects of compound (C66) of formula (I) on body weight, glucose tolerance and blood lipids in obese mice.

[0019] Figure 2 The compound (C66) of formula (I) improves renal function and pathological damage in obese mice.

[0020] Figure 3 The compound (C66) of formula (I) improves renal oxidative stress and autophagy in obese mice.

[0021] Figure 4 The effects of compound (C66) of formula (I) on body weight, blood pressure, creatinine and blood urea nitrogen in hypertensive mice.

[0022] Figure 5The compound (C66) of formula (I) improves kidney damage and oxidative stress in hypertensive mice. Detailed implementation method:

[0023] The present invention is further illustrated in the following embodiments. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0024] Example 1: The compounds of the present invention do not affect mouse body weight, glucose tolerance, or blood lipid levels.

[0025] C57L / B6 mice were randomly divided into 3 groups of 6 mice each:

[0026] Blank control group (Con group): Healthy C57L / B6 mice;

[0027] High-fat control group (HFD): Mice were fed a high-fat diet, had free access to water, and received no medication for 16 weeks;

[0028] Compound treatment group (HFD+C66 10mg / kg): Mice were fed a high-fat diet for 8 weeks. In the 9th week, the mice were administered compound (C66) of formula (I) by gavage at a dose of 10mg / kg / day for 8 consecutive weeks.

[0029] Body weight was measured in each group of mice during the period. Results are as follows: Figure 1 As shown in Figure A, there was no difference in body weight between the treatment group and the HFD group, indicating that the compound (C66) of formula (I) does not affect the weight changes in mice induced by a high-fat diet.

[0030] Two days before the end of the experiment, mice were given 1g / kg glucose orally after fasting for 12 hours. Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 minutes after oral glucose administration. A curve was plotted, and the results are shown below. Figure 1 B showed that administration of compound (C66) of formula (I) did not alleviate glucose tolerance caused by hyperlipidemia.

[0031] Eight weeks after administration, mice were sacrificed, weighed, and blood was collected. Serum levels of triglycerides (TG), total cholesterol (TCH), and high-density lipoprotein (HDL-c) were measured using a biochemical analyzer. The results are as follows: Figure 1 CE showed that administration of compound (C66) of formula (I) did not significantly alleviate the high-fat diet-induced increases in TG and TCH and decreases in HDL-c.

[0032] This indicates that compound (C66) of formula (I) has no effect on hyperlipidemia-induced weight gain, glucose tolerance, and elevated blood lipid levels, meaning that its therapeutic effect is unrelated to its lipid-lowering effect.

[0033] Example 2: The compounds of the present invention significantly improved renal function and pathological damage in hyperlipidemic mice.

[0034] Following the grouping and experimentation in Example 1, mice were sacrificed after 8 weeks of drug administration, and blood was collected to detect the levels of serum creatinine, blood urea nitrogen (BUN), and albumin (ALB). Serum creatinine, BUN, and albumin levels are important indicators of kidney disease progression; clinically, the severity of kidney disease is generally characterized by measuring serum creatinine levels. Figure 2 AC results showed that hyperlipidemia significantly increased serum creatinine, blood urea nitrogen, and albumin levels, while in the treatment group, oral compound (I) significantly reduced the increase in serum creatinine, blood urea nitrogen, and albumin (p<0.05).

[0035] Mice were grouped and tested according to Example 1. After 8 weeks of drug administration, mice in each group were sacrificed, and kidney tissue was collected. The tissue was fixed in 4% formalin, embedded in paraffin, sectioned at 5 μm, stained with hematoxylin and eosin (H&E), and examined under a microscope. The results are as follows: Figure 2 As shown in Figure D, the kidney tissue of mice in the disease control group showed significant lipid accumulation, glomerular sclerosis, enlargement, basement membrane thickening, and inflammatory cell infiltration. However, simultaneous administration of the compound of formula (I) significantly improved the symptoms of kidney tissue damage, inflammatory cell infiltration, and glomerular membrane matrix expansion in all treatment groups. This indicates that the compound (C66) of formula (I) can prevent kidney tissue dysfunction and pathological damage caused by hyperlipidemia.

[0036] Example 3: The compounds of the present invention significantly improved renal oxidative stress and autophagy in hyperlipidemic mice.

[0037] Mice were grouped and tested according to Example 1. After 8 weeks of drug administration, mice in each group were sacrificed, and kidney tissue was collected. The tissue was fixed in 4% formalin, embedded in paraffin, sectioned into 5μm sections, and subjected to 3-NT and LC3 immunohistochemical staining and microscopic examination. The results are as follows: Figure 3 As shown in Figures A and 3D, the kidney tissue of mice in the disease control group showed significant expression of the oxidative stress marker 3-NT and the formation of autophagosomes. Simultaneous administration of the compound of formula (I) significantly improved oxidative stress and autophagy in the kidney tissue of the treatment group. After homogenizing kidney tissue, total mRNA was extracted using Trizol reagent, and RT-qPCR analysis revealed that hyperlipidemia significantly increased the oxidative stress-related marker Nrf-2 in kidney tissue. Figure 3 B) and NQO-1 Figure 3C) and autophagy-related markers LC3 (3E) and Beclin 1 (3F) were inhibited. However, administration of the compound of formula (I) significantly suppressed the gene expression of Nrf-2, NQO-1, LC3, and Beclin 1 in renal tissue induced by hyperlipidemia. This indicates that the compound of formula (I) (C66) can prevent oxidative stress and autophagy in renal tissue induced by hyperlipidemia, thereby alleviating renal damage.

[0038] Example 4: The compounds of the present invention do not affect blood pressure and body weight in mice, but significantly reduce serum creatinine and blood urea nitrogen.

[0039] C57BL / 6 mice were randomly divided into 3 groups, with 6 mice in each group:

[0040] Blank control group (Con group): healthy mice;

[0041] Disease control group (Ang II group): Ang II was administered via subcutaneous micropump injection (1000 ng / kg / min), without any other medication, for 4 consecutive weeks;

[0042] Treatment group (Ang II + C66 10 mg / kg group): Ang II was administered subcutaneously via micropump injection (1000 ng / kg / min), without medication, for 4 consecutive weeks. On day 15 after disease onset, mice were administered compound (C66) of formula (I) by gavage at a dose of 10 mg / kg / day for 2 consecutive weeks.

[0043] During the period, the body weight and blood pressure of each group of mice were measured. The results are as follows: Figure 4 A and Figure 4 As shown in Figure B, there were no significant differences in body weight and blood pressure levels between the treatment group and the Ang II group, indicating that compound (C66) of formula (I) does not affect the body weight and blood pressure of hypertensive mice, and its therapeutic effect is unaffected by blood pressure values. Two weeks after administration, mice were sacrificed, blood was collected, and serum creatinine and blood urea nitrogen (BUN) levels were measured. Serum creatinine and BUN levels are important indicators of kidney disease progression; clinically, the severity of kidney disease is generally characterized by measuring serum creatinine levels. Figure 4 C and 4D results showed that hypertension significantly increased serum creatinine and blood urea nitrogen levels, while in all treatment groups, oral administration of compound (I) significantly reduced the increase in serum creatinine and blood urea nitrogen (p<0.05). These data indicate that hypertension is harmful to the kidneys, but administration of compound (I) significantly reduced hypertension-induced kidney damage, suggesting that compound (I) has a therapeutic effect on kidney lesions caused by hypertension and prevents parenchymal kidney disease.

[0044] Example 5: The compounds of the present invention significantly improved renal lesions and oxidative stress in hypertensive mice.

[0045] Following the grouping and experimentation in Example 4, mice in each group were sacrificed after two weeks of drug administration. Kidney tissue was collected, fixed in 4% formalin, embedded in paraffin, sectioned at 5 μm, and subjected to hematoxylin and eosin (H&E) and 3-NT immunohistochemical staining and microscopic examination. The results are as follows: Figure 5 As shown in AB, the kidney tissue of mice in the disease control group exhibited significant structural abnormalities (including glomerular sclerosis, enlargement, and thickening of the basement membrane, as well as severe inflammatory cell infiltration) and oxidative stress. Simultaneous administration of the compound of formula (I) significantly improved symptoms such as kidney tissue damage, inflammatory cell infiltration, glomerular membrane matrix expansion, and oxidative stress in all treatment groups. After homogenizing kidney tissue, total mRNA was extracted using Trizol reagent, and RT-qPCR analysis revealed that hypertension significantly increased the levels of Nrf-2, an oxidative stress-related marker, in kidney tissue. Figure 5 C) and NQO-1 Figure 5 The expression of Nrf-2 and NQO-1 in kidney tissue induced by hypertension was significantly inhibited by the compound of formula (I), while the expression of Nrf-2 and NQO-1 in kidney tissue induced by hypertension was significantly inhibited by the compound of formula (I). This indicates that the compound of formula (I) (C66) can prevent the exacerbation of kidney damage and oxidative stress caused by hypertension.

Claims

1. The use of (2E,6E)-2,6-bis(2-(trifluoromethyl)phenylmethylene)cyclohexanone or a pharmaceutically acceptable salt thereof in the preparation of pharmaceuticals, characterized in that, The aforementioned medication is used to treat or prevent kidney-related diseases; The kidney-related diseases mentioned are kidney tissue dysfunction and pathological damage caused by hyperlipidemia, or kidney tissue oxidative stress and autophagy caused by hyperlipidemia.

2. The use of (2E,6E)-2,6-bis(2-(trifluoromethyl)phenylmethylene)cyclohexanone or a pharmaceutically acceptable salt thereof in the preparation of a drug according to claim 1, characterized in that, The symptoms of kidney tissue dysfunction and pathological damage include lipid accumulation in the kidney tissue.

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