WO2021036495A1

WO2021036495A1 - Novel phenylacetic acid derivative, preparation method thereof and use thereof as drug

Info

Publication number: WO2021036495A1
Application number: PCT/CN2020/099650
Authority: WO
Inventors: 李政; 张陆勇; 刘冰
Original assignee: 广东药科大学
Priority date: 2019-08-29
Filing date: 2020-07-01
Publication date: 2021-03-04
Also published as: CN112441916A

Abstract

The present invention relates to a novel deuterated 3-pentylphenylacetic acid derivative represented by general formula (I), a preparation method thereof, and a use of a pharmaceutical composition containing the derivative in preparation of a drug for preventing or/and treating organ fibrosis-associated diseases. The compound of the present invention has more excellent drug metabolism properties and anti-organ fibrosis effect in vivo, and can be applied to the preparation of drugs for preventing and/or treating fatty liver, hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, Alström syndrome and other organ fibrosis associated diseases, and has a wide application prospect.

Description

Novel phenylacetic acid derivative, its preparation method and its use as medicine

Technical field

The invention relates to a deuterated 3-pentylphenylacetic acid derivative, a preparation method and application thereof, and belongs to the technical field of medicine. The structure of the deuterated derivative involved in the present invention is unique and novel in this field.

Background technique

Organ fibrosis refers to the accumulation of infection, inflammation, fat and other substances in the living body caused by various reasons such as microorganisms, chemical substances, immune responses, eating habits, environment, genetic background and other reasons (including unknown causes). After the degeneration of tissues or cells causes organ damage, necrosis, etc., it cannot fully regenerate, and connective tissue proliferation and fibrosis occur. It is known to occur in various organs such as the liver, lungs, heart, kidney, cranial nervous system, and many organs and tissues such as muscles, bones, and skin. Liver cirrhosis is a typical pathological state of fibrosis in the liver. It is the chronic development of liver disease caused by various reasons (such as drug-induced liver injury, fatty liver, chronic hepatitis, etc.) to the final morbid state, due to the reduction of functional liver cells and fibrosis Tissue hyperplasia, liver function is significantly reduced. There are about 20 million patients with liver cirrhosis in the world, but there is no effective treatment drug yet. At present, the main preventive method is how to delay the development of chronic liver disease to cirrhosis through various symptomatic therapies. In pulmonary fibrosis, in addition to fibrosis caused by lung injury accompanied by pneumonia such as interstitial pneumonia, there is also idiopathic pulmonary fibrosis of unknown cause. The disease has symptoms such as cough, chest pain, and dyspnea, which seriously affects patients Quality of life and poor prognosis. Myocardial fibrosis is a disease in which the tissues of the myocardium and heart valves become fibrotic. The primary disease is cardiomyopathy and valvular disease caused by coronary circulatory failure, infection or immune response, etc., and can progress to a state of heart failure. In the kidney, if fibrosis is caused in the kidney tissue along with the development of chronic kidney disease, it will cause the kidney function to deteriorate rapidly and reach an irreversible state. In short, the development of organ fibrosis is a serious disease that causes irreversible deterioration of the function of the organ and tissue and has a very poor prognosis. However, there is still a lack of effective drugs. Liver transplantation is the only effective treatment method for liver cirrhosis, but liver transplantation has many problems such as insufficient organ donors, high medical costs, and health risks to the donor in the case of living donor liver transplantation. Therefore, the development of drugs for organ fibrosis has become a research hotspot in this field, in order to bring more economical, safe and effective new drugs to patients with organ fibrosis.

Pirfenidone has anti-inflammatory, anti-fibrotic and antioxidant properties, and can significantly delay the rate of forced expiratory vital capacity (FVC) decline. It was approved for the treatment of idiopathic pulmonary fibrosis (IPF) in 2008 and is recommended for light It is not clear whether IPF patients with moderate pulmonary dysfunction and severe pulmonary dysfunction can benefit. Nintanib is a VEGFR/FGFR/PDGFR inhibitor, which can significantly reduce the absolute value of FVC decline in patients with IPF and relieve the disease process to a certain extent. It was approved for IPF in the United States and the European Union in 2014 and 2015, respectively. Like pirfenidone, this product is recommended for patients with mild and moderate pulmonary dysfunction. Whether severe patients can benefit or not requires further research. However, pirfenidone and nintedanib have greater side effects and poor patient tolerance. In recent years, a series of clinical studies have shown that PBI-4050 (WO2012097428) can significantly improve the fibrosis of various organs with less side effects (Nat Rev Drug Discov, 2017, 16, 755-772 and Eur Respir J, 2019, 53, 1800663), However, its benzylic site is easily oxidized and metabolized in the body, thereby increasing the metabolic burden of the liver, affecting its pharmacokinetic properties, and ultimately affecting the anti-fibrosis effect in the body.

The present invention uses a deuteration strategy to block the potential metabolic sites of PBI-4050, which has better drug metabolism properties and anti-organ fibrosis effects in vivo. Therefore, the deuterated PBI-4050 and its pharmaceutically acceptable salts can be potentially used to treat or prevent organ fibrosis-related diseases, and have broad development prospects.

Summary of the invention

In view of the defects of PBI-4050 that is easy to metabolize and unmet clinical needs in the prior art, the purpose of the present invention is to provide deuterated PBI-4050 derivatives and their medical applications, and provide a solution for the prevention or/and treatment of organ fibrosis-related diseases. Class of new potential drugs.

The deuterated PBI-4050 derivative of the present invention contains an effective amount of the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof:

among them:

R ₁ , R ₂ and R ₃ are each independently selected from hydrogen and deuterium, and _{at least one of the R 1} , R ₂ and R ₃ groups is a deuterium atom;

More preferred compounds of the invention include, but are not limited to:

2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)acetic acid (I-1);

2-(3-(pentyl-1,1-D ₂ )phenyl)-2,2-D ₂ acetic acid (I-2);

2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)-2,2-D ₂ acetic acid (I-3);

Sodium 2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)acetate (I-4).

The pharmaceutically acceptable salt involved in the present invention is a sodium salt, a potassium salt or a lithium salt.

Another aspect of the present invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound or a pharmaceutically acceptable salt thereof and a suitable carrier, diluent or excipient.

The present invention also relates to the use of the compound or its pharmaceutically acceptable salt or its pharmaceutical composition in the preparation of a medicament for the prevention or/and treatment of organ fibrotic diseases, wherein the organ fibrosis occurs in the digestive organs, respiratory organs, Fibrosis of circulatory organs, genitourinary organs, motor organs, cranial nervous system, endocrine organs, or skin; including prevention or/and treatment of fatty liver, liver fibrosis, pulmonary fibrosis, kidney fibrosis, heart fibrosis,

Syndrome and other organ fibrosis-related diseases in medicine for at least one disease.

The present invention can be measured by using the test system described below.

The following biological test examples describe the present invention.

The experimental methods of the specific conditions in the test examples of the present invention are usually in accordance with the conventional conditions or in accordance with the conditions recommended by the commercial manufacturer. The reagents without specific sources are commonly used reagents purchased on the market.

Test Example 1 Study on the stability of the compound of the present invention on human liver microsomes

Prepare the mother liquor of the test compound with DMSO, and dilute it with 0.1% BSA solution to the concentration to be tested. Human liver microsomes were incubated in a 96-well plate containing 0.1M Tris buffer (pH 7.4), 5.0mM MgCl ₂ solution, 0.005% BSA, and 1.0mM NADPH at 37°C. The final concentration of microsomal protein per well was 0.5 mg/mL, after incubating for 5 minutes, add the test drug, and add methanol to stop the reaction after 0, 10, 20, 30, 60, and 90 minutes. Determine the concentration of the compound in the sample by LC-MS/MS, and calculate the half-life of the compound. See Table 1 for the stability results of liver microsomes of the example compounds.

Table 1: Stability of compounds to human liver microsomes

Conclusion: All the compounds of the present invention have stronger stability to human liver microsomes than PBI-4050, and have the potential to reduce the dosage of drugs and improve the efficacy of drugs in vivo.

Test Example 2 The in vivo anti-hepatic fibrosis effect of the compound of the present invention can be measured by using the following measurement system:

8-week-old C57BL/6 mice, male, were randomly divided into 6 groups with 6 mice in each group. They were given 2mL/kg carbon tetrachloride olive oil solution (10% content) twice a week. The model was built for 60 days. During the model period, the blank control group (blank vehicle: 0.5% sodium carboxymethyl cellulose solution) and the test compound group (200 mg/kg) were given the blank vehicle and the test compound by intragastric administration once a day for 60 consecutive days. The mice were sacrificed on the 61st day, serum was collected, and serum AST and ALT levels were determined by an automatic biochemical analyzer; the liver was removed, and the content of hydroxyproline in the liver was determined by alkaline hydrolysis, and the liver fibrosis of mice was observed by Masson staining status. AST, ALT and hydroxyproline levels are shown in Table 2. See Figure 1 for the results of Masson staining.

Table 2: Effects of compounds on serum AST, ALT and liver hydroxyproline levels (

n=6)

Note: *P≤0.05 is the Student’s t-test result relative to the PBI-4050 group.

The results show that the compound of the present invention and its medicinal sodium salt I-4 can significantly improve the liver function of the mouse model of liver fibrosis and reduce the level of hydroxyproline in the liver; Masson staining (figure 1) shows that the model control group The fibrous septa of the liver lobules were formed, and the fibrous deposition around the central vein and the portal area increased. The degree of liver fibrosis in the administration group was significantly improved, showing a good anti-liver fibrosis effect. Among them, the AST levels of compounds I-1, I-3 and I-4 were significantly lower than those of the PBI-4050 group, and the ALT and hydroxyproline levels of the I-3 group were significantly lower than those of the PBI-4050 group. Medicinal value.

Test Example 3 The in vivo anti-kidney fibrosis effect of the compound of the present invention can be measured by using the measurement system described below:

8-week-old C57BL/6 mice, males, were randomly divided into 6 groups, each with 6 mice. The renal fibrosis model was prepared by feeding with 0.25% adenine feed for four weeks. After being given adenine feed for one week, the blank control group was started. The test compound group (200mg/kg) was intragastrically administered with blank vehicle and test compound once a day. After three weeks of continuous administration, the mice were sacrificed, serum was collected, and serum urea nitrogen (BUN) and blood creatinine (Scr) levels were determined; Take the kidney to observe the fibrosis state of the mouse kidney by Masson staining. See Table 3 for BUN and Scr levels. See Figure 2 for the results of Masson staining.

Table 3: Effects of compounds on BUN and Scr levels (

n=6)

Note: **P≤0.01 is the result of Student’s t test relative to the blank control group.

The results show that the test compound can significantly improve the renal function indicators of the renal fibrosis model. Masson staining (figure 2) shows that the model control group has dilated renal tubules, thickened glomerular basement membrane, and increased fibrous proliferation. Renal fibrosis of the group was significantly improved, and the compound of the present invention showed a better anti-renal fibrosis effect.

Description of the drawings

Figure 1: Masson staining results of liver fibrosis model induced by _{CCl 4}

Figure 2: Masson staining results of adenine-induced renal fibrosis model kidney

detailed description

In the following, the present invention will be further described in conjunction with the embodiments. It should be noted that the following embodiments are only for illustration, but not for limiting the present invention. Various changes made by those skilled in the art based on the teachings of the present invention should fall within the protection scope required by the claims of this application.

Example 1

2-(3-(Pentyl-1,1,2,2-D ₄ )phenyl)acetic acid (I-1)

Step 1: In the sealed tube, add Pd(PPh ₃ ) ₂ Cl ₂ (0.6g, 0.78mmol), 1-pentyne (5.4g, 78.6mmol), and (3-bromophenyl) methyl acetate (6g , 26.2mmol) and tetrabutylammonium fluoride hydrate (20.6g, 78.6mmol), mixed well and heated at 80°C for 3h. The mixture was added with 50 mL of water, and extracted with ethyl acetate (20 mL×3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the organic solvent was evaporated under reduced pressure. Purification by column chromatography (ethyl acetate/petroleum ether, 5:95, v/v) gave methyl (3-(pentyn-1-yl)phenyl)acetate (3.6 g, 65%) as a pale yellow oil.

¹ H NMR(300MHz,DMSO-d ₆ )δ7.33-7.28(m,3H), 7.27-7.22(m,1H), 3.70(s,2H), 3.63(s,3H), 2.41(t,J =7.0Hz, 2H), 1.58 (h, J = 7.2Hz, 2H), 1.02 (t, J = 7.3Hz, 3H).

Step 2: Mix 10% Pd/C (0.36g) with D ₂ O (30 mL), after replacing hydrogen, react at room temperature for 24 hours, add (3-(pentyn-1-yl)phenyl)acetate to the reaction flask Ester (3.6g) in methanol dilution, continue to react at room temperature for 6h, the reaction solution was filtered with diatomaceous earth, the filter cake was washed with ethyl acetate, the filtrate was collected, 20mL of water was added, ethyl acetate extracted, and the organic phases were combined with anhydrous sodium sulfate Dry, filter, and evaporate the organic solvent under reduced pressure. Purification by column chromatography (ethyl acetate/petroleum ether, 3:97, v/v) to obtain methyl 2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)acetate (3.2 g, 89%).

Step 3: Dissolve methyl 2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)acetate (3.2g, 14.3mmol) in a mixture of _{THF/CH 3} OH/H _{2 O} LiOH (1.7g, 71.4mmol) was added to the solvent. After the addition, the reaction was carried out at room temperature for 5h. The solvent was evaporated under reduced pressure, acidified with 1N HCl, and extracted with ethyl acetate. The combined organic phases were dried with anhydrous sodium sulfate, filtered, and reduced under reduced pressure. The organic solvent is evaporated. The residue was purified by column chromatography (ethyl acetate/petroleum ether, 1:3, v/v) to obtain 2-(3-(pentyl-1,1,2,2-D ₄ )phenyl) as a white solid Acetic acid was 2.9 g, and the yield was 91%.

¹ H NMR (300MHz, CD ₃ OD) δ: 7.23--7.19 (m, 1H), 7.09--7.04 (m, 3H), 3.57 (s, 2H), 1.36--1.32 (m, 4H), 0.88 (t, J=7.2Hz,3H).ESI-MS m/z:209.1[MH] ^- .

Example 2

2-(3-(Pentyl-1,1-D ₂ )phenyl)-2,2-D ₂ acetic acid (I-2)

Add m-pentylphenylacetic acid (3g), 10% palladium on carbon (0.3g) and heavy water (25mL) to a round bottom flask. After hydrogen replacement, heat at 50°C for 72h. The reaction solution is filtered with diatomaceous earth. Acetic acid The filter cake was washed with ethyl acetate, the filtrate was collected, 20 mL of water was added, and the mixture was extracted with ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the organic solvent was evaporated under reduced pressure. The residue was purified by column chromatography (ethyl acetate/petroleum ether, 1:3, v/v) to obtain 2.6 g of a white solid with a yield of 86%.

¹ H NMR (300MHz, CD ₃ OD)δ: 7.25–7.20(m,1H), 7.08–7.04(m,3H), 1.62(t,J=7.4Hz,2H), 1.37–1.32(m,4H) ,0.87(t,J=7.1Hz,3H).ESI-MS m/z:209.1[MH] ^- .

Example 3

2-(3-(Pentyl-1,1,2,2-D ₄ )phenyl)-2,2-D ₂ acetic acid (I-3)

In a round bottom flask, I-1 (2g), 10% palladium on carbon (0.2g) and heavy water (20mL) were sequentially added. After hydrogen replacement, the reaction was heated at 50°C for 72h. The reaction solution was suction filtered with diatomaceous earth and ethyl acetate Wash the filter cake, collect the filtrate, add 20 mL of water, and extract with ethyl acetate. The combined organic phases are dried over anhydrous sodium sulfate, filtered, and the organic solvent is evaporated under reduced pressure. The residue was purified by column chromatography (ethyl acetate/petroleum ether, 1:3, v/v) to obtain 1.6 g of a white solid with a yield of 80%.

¹ H NMR(300MHz, CD ₃ OD)δ: 7.28–7.22(m,1H), 7.09–7.04(m,3H), 1.36–1.32(m,4H), 0.89(t,J=7.3Hz,3H) .ESI-MS m/z:211.1[MH] ^- .

Example 4

Sodium 2-(3-(pentyl-1,1,2,2-D ₄ )phenyl)acetate (I-4)

In the mixed solution of ethanol and water (4:1, v/v), I-1 (2g, 3.5mmol) and sodium bicarbonate (0.8g, 3.5mmol) were sequentially added. The reaction mixture was stirred overnight at room temperature. The solvent was evaporated under reduced pressure, the obtained white sticky solid was dissolved in water, and the solution was lyophilized to obtain 1.9 g of white solid with a yield of 86%.

Example 5

Tablets containing active agent I-4:

The active ingredients, pregelatinized starch and microcrystalline cellulose are sieved, mixed thoroughly, added to the polyvinylpyrrolidone solution, mixed, soft materials are made, sieved, wet granules are made, dried at 50-60℃, and the carboxymethyl starch The sodium salt, magnesium stearate and talc are sieved and added to the above granules to form a tablet.

Claims

A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof:

among them:

R 1 , R 2 and R 3 are each independently selected from hydrogen and deuterium, and at least one of the R 1 , R 2 and R 3 groups is a deuterium atom.
The compound of general formula (I) as defined in claim 2 or a pharmaceutically acceptable salt thereof, said compound being selected from:

2-(3-(pentyl-1,1,2,2-D 4 )phenyl)acetic acid;

2-(3-(pentyl-1,1-D 2 )phenyl)-2,2-D 2 acetic acid;

2-(3-(pentyl-1,1,2,2-D 4 )phenyl)-2,2-D 2 acetic acid;

Sodium 2-(3-(pentyl-1,1,2,2-D 4 )phenyl)acetate.
The pharmaceutically acceptable salt of the compound of general formula (I) according to any one of claims 1-2 is a sodium salt, a potassium salt or a lithium salt.
A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and a suitable carrier or excipient.
Use of the compound defined in any one of claims 1 to 4, or a pharmaceutically acceptable salt or pharmaceutical composition thereof, in the preparation of a medicine for preventing or/and treating organ fibrotic diseases, wherein organ fibrosis occurs in digestion Fibrosis of organs, respiratory organs, circulatory organs, urogenital organs, motor organs, cranial nervous system, endocrine organs, or skin.
The compound defined in any one of claims 1 to 4, or a pharmaceutically acceptable salt or pharmaceutical composition thereof, is used in the preparation of prevention or/and treatment of fatty liver, liver fibrosis, pulmonary fibrosis, kidney fibrosis, heart fibrosis,
Syndrome and other organ fibrosis-related diseases in medicine for at least one disease.