New bezafibrate salt and application thereof

Abstract

The invention discloses a novel bezafibrate salt and an application thereof. The novel bezafibrate salt comprises a bezafibrate nicotinamide salt, a bezafibrate NMNAM salt, a bezafibrate choline salt, a bezafibrate lysine salt, a bezafibrate methionine methyl ester salt and a bezafibrate methionine ethyl ester salt. The new salts not only significantly improve the bioavailability of bezafibrate, but also show great simplicity and high efficiency in the preparation method, and provide a new choice for drug development and clinical application.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and in particular to a new salt of bezafibrate and its applications. Background Art

[0002] Bezafibrate belongs to the fibrate lipid-lowering drugs, which can significantly reduce triglycerides and low-density lipoproteins, increase the level of high-density lipoproteins, delay the progression of coronary atherosclerotic lesions, and reduce the occurrence of coronary heart disease events. In recent years, studies have found that bezafibrate can also inhibit the synthesis of bile acids and has the effect of treating primary biliary cholangitis. Bezafibrate is rapidly absorbed orally, with a Tmax of about 2 h, but is excreted quickly. After single oral administration of the ordinary preparation of bezafibrate to humans, the elimination half-life T1 / 2β is about 1.5 - 2.0 hours, and 95% of the drug is excreted through urine, and about 50% is excreted in the form of the original drug, with poor bioavailability. The ordinary tablets of bezafibrate need to be orally administered three times a day, 200 mg each time, to achieve better therapeutic effects. Bezafibrate is a drug that needs to be taken for a long time. The three-times-a-day administration has poor patient compliance and affects the efficacy. The dose of 600 mg per day is also prone to produce toxic and side effects.

[0003] In order to improve the pharmacokinetic properties of bezafibrate and increase its bioavailability, many studies on bezafibrate have been carried out in various aspects such as formulation and molecular structure modification. In terms of formulation: freeze-drying granulation, preparation of disintegrating tablets, solid dispersions, research and development of sustained-release preparations, ultrafine powder of the active ingredient, preparation of nanofluids, electrosprayed nanospheres, nano-liposomes, preparation of osmotic pump tablets using semi-solid 3D printing technology, and use of new pharmaceutical excipients to improve the bioavailability of drugs, etc. In terms of molecular structure modification, mainly prodrugs are prepared. For example, WO2019168842 prepared N-methylamide derivatives, and WO2012145899 prepared bezafibrate ester prodrugs. However, at present, only the sustained-release tablets have been approved for marketing, with a daily dosage of 400 mg, once a day. There is still a large room for improvement in the pharmacokinetic properties of bezafibrate. Summary of the Invention

[0004] The object of the present invention is to provide a new salt of bezafibrate and its applications in view of the deficiencies in the prior art.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] The first aspect is to provide a bezafibrate salt represented by the structural formula (I),

[0007]

[0008] wherein, M + is a quaternary ammonium salt or AH + , and A is selected from an amino acid or its ester derivative.

[0009] Furthermore, A is selected from L-amino acids, D-amino acids, or a mixture of L- and D-amino acids.

[0010] More preferably, the amino acid is selected from natural amino acids, preferably amino acids present in the human body. Most preferably, lysine (Formula (II)), arginine, and methionine methyl ester (Formula (III)):

[0011]

[0012] Among them, R is a C1-C8 alkyl group, preferably a C1-C4 normal alkyl group, namely methyl, ethyl, n-propyl, and n-butyl.

[0013] Furthermore, M + is selected from choline (Formula (IV)) or N-methyl nicotinamide (abbreviated as NMNAM, Formula (V)):

[0014]

[0015]

[0016] The above-mentioned new bezafibrate salts provided by the present invention can be directly reacted with corresponding bases such as lysine and choline hydroxide using bezafibrate. The reaction ratio (molar ratio) of bezafibrate to the base is 1:(0.2 - 10), preferably 1:(0.5 - 2), and most preferably 1:1.

[0017] The reaction temperature is -20°C - 140°C, preferably 0°C - 100°C, and most preferably room temperature to 80°C. The reaction solvent is not limited, and preferably water and solvents miscible with water, including water, C1-C4 alcohols, C3-C8 ketones, tetrahydrofuran, methyltetrahydrofuran, and their mixtures.

[0018] The above-mentioned new bezafibrate salts provided by the present invention can also be prepared by first preparing the sodium salt or potassium salt of bezafibrate and then reacting it with corresponding coordination salts such as NMNAM hydrochloride, D-methionine methyl ester hydrochloride, and L-methionine ethyl ester hydrochloride. The reaction ratio (molar ratio) of bezafibrate to the base is 1:(0.2 - 10), preferably 1:(0.5 - 2), and most preferably 1:1.

[0019] The reaction temperature is -20°C - 140°C, preferably 0°C - 100°C, and most preferably room temperature to 80°C. The reaction solvent is not limited, and preferably water and solvents miscible with water, including water, C1-C4 alcohols, C3-C8 ketones, tetrahydrofuran, and methyltetrahydrofuran, and their mixtures.

[0020] The second aspect is to provide a pharmaceutical composition with the above-mentioned bezafibrate salt as the main active ingredient.

[0021] Further, the pharmaceutical composition is administered by injection, and the selected dosage form is sterile powder injection, small volume injection or large volume infusion.

[0022] Further, the pharmaceutical composition is administered by oral route, and the selected dosage forms are tablets, coated tablets, capsules, granules, oral solutions.

[0023] For the pharmaceutical composition provided by the present invention, the active ingredient does not need to be micronized or ultramicronized, and only needs to be conventionally pulverized to meet the requirements of the pharmaceutical preparation process.

[0024] In addition to the bezafibrate salts provided by the present invention, the pharmaceutical composition provided by the present invention preferably adds a certain amount of pharmaceutical preparation excipients. For injection preparations, solubilizers, preservatives, chelating agents, pH regulators, osmotic pressure regulators, etc. are preferably added. For oral preparations, diluents, disintegrants, binders, drug release rate regulators, coating agents, etc. are preferably added. The pharmaceutical excipients can be one kind, and preferably two or more kinds are added.

[0025] The definitions and types of the above-mentioned solubilizers, preservatives, chelating agents, pH regulators, osmotic pressure regulators, diluents, disintegrants, binders, drug release rate regulators, coating agents, etc. are the same as those known to professionals in this field, and they are all pharmaceutical excipients approved by the drug regulatory department for use in pharmaceutical preparations.

[0026] The third aspect is to provide the application of the above-mentioned bezafibrate salts in the preparation of drugs for regulating blood lipids, preferably for reducing high triglycerides and / or increasing high-density lipoprotein.

[0027] The fourth aspect is to provide the application of the above-mentioned bezafibrate salts in the preparation of drugs for treating hepatobiliary diseases. The hepatobiliary diseases are preferably cholangitis, cholecystitis, gallstones, cholestasis, fatty liver. Most preferably, it is used for treating primary biliary cholangitis and cholestasis diseases.

[0028] When the bezafibrate salts and their pharmaceutical compositions provided by the present invention are used for regulating blood lipids and treating hepatobiliary diseases, they can be used simultaneously with other drugs as needed. It is preferably used in combination with statins or ursodeoxycholic acid.

[0029] By adopting the above technical solutions, compared with the prior art, the present invention has the following technical effects:

[0030] The novel bezafibrate salts of the present invention specifically include, but are not limited to, bezafibrate NMNAM salt, bezafibrate choline salt, bezafibrate lysine salt, bezafibrate methionine methyl ester salt and bezafibrate methionine ethyl ester salt; these novel salts significantly improve the bioavailability and solubility of bezafibrate in water, and there is no need for ultramicronization or preparation of nanofluids when preparing pharmaceutical compositions, which simplifies the process, improves the curative effect, and provides new options for drug development and clinical applications. Detailed embodiments

[0031] The present invention will be further described below in conjunction with specific embodiments, but it is not intended to limit the present invention. It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other.

[0032] Example 1 Preparation of bezafibrate NMNAM salt

[0033]

[0034]

[0035] Put 3.00 g of bezafibrate, 1.5 g of NMNAM hydrochloride, 0.338 g of sodium hydroxide, and 25 ml of 50% ethanol into a reaction flask, stir, and heat until all the solids are dissolved and clear. Cool to room temperature and place in the refrigerator overnight. Solids precipitate out, filter by suction, filter out the solids, and dry to obtain the product with a yield of about 50%.

[0036] 1H NMR (400 MHz, D2O) σ 1.41 (S, 6H), 2.79 (T, 2H), 3.52 (T, 2H), 4.39 (S, 3H), 6.78 (D, 2H), 7.11 (D, 2H), 7.39 (D, 2H), 7.48 (D, 2H), 8.08 (T, 1H), 8.80 (D, 1H), 8.87 (D, 1H), 9.18 (S, 1H)

[0037] Example 2 Preparation of bezafibrate choline salt

[0038]

[0039] Put 3.00 g of bezafibrate and 50 ml of absolute ethanol into a reaction flask and stir.

[0040] Dissolve 2.3 g of choline hydroxide aqueous solution (44%) in 10 ml of absolute ethanol, and drop it into the above reaction solution. While dropping bezafibrate, it dissolves. After dropping, it is immediately dissolved and clear. Continue to stir for 2 hours, concentrate under reduced pressure to dryness to obtain a gum, add 30 ml of isopropanol, stir, solids precipitate out, heat until all the solids are dissolved, and cool to precipitate crystals. Filter by suction, filter out the solids and dry to obtain 1.12 g of the product.

[0041] 1H NMR (400 MHz, D2O) σ 1.39 (S, 6H), 2.75 (T, 2H), 3.09 (S, 9H), 3.41 (T, 2H), 3.47 (T, 2H), 3.92 (T, 2H), 6.76 (D, 2H), 7.08 (D, 2H), 7.33 (D, 2H), 7.44 (D, 2H)

[0042] Preparation of Bezafibrate Lysine Salt

[0043]

[0044] Dissolve 1.00 g of bezafibrate and 0.405 g of lysine in 14 ml of methanol in a reaction flask and stir. It will become clear after a few minutes. Concentrate to dryness to obtain a gummy substance. Add absolute ethanol twice to obtain 1.3 g of a white powdery product.

[0045] 1H NMR(400MHz,D2O)σ1.38 - 1.42(m,2H),1.40(S,6H),1.62(m,2H),1.80 - 1.84(m,2H),2.79(T,2H),2.94(T,2H),3.52(T,2H),3.66 - 3.80(m,1H),6.77(D,2H),7.11(D,2H),7.40(D,2H),7.49(D,2H)

[0046] Preparation of Bezafibrate Methionine Methyl Ester Salt

[0047]

[0048] Dissolve 0.55 g of D - methionine methyl ester hydrochloride in 5 ml of water, add 0.25 g of sodium bicarbonate, stir for 0.5 hour, extract three times with 5 ml of dichloromethane each time. Combine the extracts, dry with anhydrous magnesium sulfate, filter off the magnesium sulfate, add 1.00 g of bezafibrate to the filtrate and stir for 1 hour. Concentrate under reduced pressure to dryness to obtain a gummy substance. Dissolve the gummy substance in acetic acid, and then a large amount of solid will precipitate. Filter by suction, filter out the solid, and dry to obtain 1.3 g.

[0049] 1HNMR(400MHz,DCCl3)σ1.58(s,6H),1.86 - 1.92(m,2H),2.07(s,3H),2.62 - 2.64(m,2H),2.88(t,2H),3.69(t,2H),3.67 - 3.70(m,1H),3.75(s,3H),6.15(br,1H),6.89(d,2H),7.12(d,2H),7.38(d,2H),7.62(d,2H),8.86(br,3H)

[0050] Preparation of Bezafibrate Methionine Ethyl Ester Salt

[0051]

[0052] Dissolve 0.56 g of L-methionine ethyl ester hydrochloride in 5 ml of water, add 0.25 g of sodium bicarbonate, stir for 0.5 hour, extract three times with 5 ml of dichloromethane each time, combine the extracts, dry with anhydrous magnesium sulfate, filter off the magnesium sulfate, add 1.00 g of bezafibrate to the filtrate, and stir for 1 hour. Concentrate under reduced pressure to dryness to obtain a gum. Dissolve the gum in ethyl acetate, and then a large amount of solid precipitates. Filter by suction, filter out the solid, and dry to obtain 1.4 g.

[0053] 1HNMR(400MHz,DCCl3)σ1.28(t,3H),1.57(s,6H),1.86 - 1.87(m,2H),2.10(s,3H),2.62(br,2H),2.86(t,2H),3.64(t,2H),3.43(br,1H),4.20(q,2H),6.21(s,1H),6.88(d,2H),7.10(d,2H),7.37(d,2H),7.63(d,2H),8.80(br,3H)

[0054] Comparative Example 1 Preparation of Sodium Bezafibrate

[0055] Place 3.00 g of bezafibrate, 0.338 g of sodium hydroxide, and 75 ml of methanol in a reaction flask and stir until all the solids dissolve. Concentrate under reduced pressure to dryness to obtain sodium bezafibrate.

[0056]

[0057] Comparative Example 2 Preparation of Calcium Bezafibrate

[0058] Place 1.00 g of bezafibrate, 0.112 g of sodium hydroxide, and 15 ml of water in a reaction flask and stir for a moment until it dissolves clearly.

[0059] Dissolve 0.155 g of calcium chloride in 5 ml of water, drop it into the above reaction solution, and solid precipitates while dropping. After dropping, continue to stir for 2 hours, filter by suction, filter out the solid, and dry to obtain calcium bezafibrate (weighing 1 g).

[0060] Comparative Example 3 Preparation of Magnesium Bezafibrate

[0061] Place 1.00 g of bezafibrate, 0.112 g of sodium hydroxide, and 15 ml of water in a reaction flask and stir for a moment until it dissolves clearly.

[0062] Dissolve 0.15 g of magnesium chloride in 5 ml of water, drop it into the above reaction solution, and solid precipitates while dropping. After dropping, heat, and part of the solid dissolves. Continue to stir for 2 hours, cool, and solid precipitates. Filter by suction, filter out the solid, and dry to obtain magnesium bezafibrate (weighing 0.90 g).

[0063] Bioavailability Study of Verification Example 1

[0064] Animals

[0065] Sprague-Dawley rats, male, weighing 200 - 250 g. Fast for 12 hours before the start of the experiment, but can drink water freely.

[0066] Drugs

[0067] Control drug: Bezafibrate. Since bezafibrate is insoluble in water, it is made into a suspension with a concentration of 0.6 mg / ml using a 0.5% methylcellulose saline solution. Test drug: The bezafibrate salts prepared in Examples 1 - 5 and Comparative Example 1 above. It is made into a solution equivalent to 0.6 mg / ml of bezafibrate with normal saline. The specific concentrations are shown in Table 1:

[0068] Table 1

[0069]

[0070]

[0071] Grouping and Sample Collection and Preservation

[0072] The rats are randomly divided into seven groups, with 8 rats in each group. Group 1 orally administers 1 ml of bezafibrate suspension, equivalent to a bezafibrate dose of 3 mg / kg. Groups 2 - 7 sequentially orally administer 1 ml of sodium bezafibrate, NMNAM salt, choline salt, lysine salt, ethyl methionine salt, and methyl methionine salt, respectively, equivalent to a bezafibrate dose of 3 mg / kg. All formulations are freshly prepared before drug administration.

[0073] At predetermined time points of 5, 15, and 30 minutes, 1, 2, 3, 4, 6, 8, 10, 12, 18, 24, 48, and 72 hours after drug administration, blood samples are collected from the retro-orbital vein. The blood samples are collected in tubes pre-added with 0.5 mL of sodium heparin, and centrifuged (3000 rpm) for 10 minutes to obtain plasma. All plasma samples are stored in a freezer at -20°C before analysis.

[0074] Extraction and Separation of Plasma Samples

[0075] Take plasma samples (50 μL), add 10 μL of 10% perchloric acid solution, shake for 10 minutes, centrifuge for 10 minutes, take 10 μL of the supernatant and inject it into a high-performance liquid chromatograph to measure the peak area of bezafibrate, and calculate the blood concentration at each time point by the external standard method.

[0076] High-Performance Liquid Chromatography Determination Conditions

[0077] Mobile phase: composed of methanol - 0.01 mol / L acetate buffer (pH 4.0) - 0.5 mol / L tetrabutylammonium bromide (74:26:0.2); Flow rate: 1.0 ml / min; Chromatographic column: Agilent ZORBAX ODS C18 (5.0 μm, 250 nm X 4.6 mm); Detection wavelength: 232 nm; Column temperature: 40 °C.

[0078] Data processing and statistical analysis

[0079] The data was processed using the 3P97 pharmacokinetic program, and the AUC was compared by the double one-sided t-test method for bioavailability evaluation. The pharmacokinetic parameters are shown in Table 2 below.

[0080] Table 2

[0081]

[0082]

[0083] As can be seen from Table 2, bezafibrate sodium has little effect on the pharmacokinetic parameters of bezafibrate. Bezafibrate NMNAM salt, choline salt, lysine salt, and methionine salt have obvious effects on pharmacokinetics, and the bioavailability is significantly improved.

[0084] Verification Example 2 Solubility test

[0085] Weigh a certain amount of bezafibrate and an equivalent amount of bezafibrate salt, shake with water, and observe the dissolution of the sample. The test results are shown in Table 3.

[0086] Table 3

[0087]

[0088] As can be seen from Table 3, the solubility of bezafibrate sodium salt, NMNAM salt, choline salt, lysine salt, and methionine ester salt in water is greatly improved compared to bezafibrate. The calcium salt and magnesium salt of bezafibrate have little effect.

[0089] The above is only a preferred embodiment of the present invention, and it does not limit the implementation manner and protection scope of the present invention. For those skilled in the art, it should be realized that all equivalent replacements and obvious changes made by using the content of the present invention specification should be included in the protection scope of the present invention.

Claims

1. A bezafibrate salt as shown in structural formula (I), characterized in that: Among them, M + For quaternary ammonium salt or AH + , A is selected from amino acids or their ester derivatives.

2. The bezafibrate salt according to claim 1, characterized in that The A is selected from lysine, arginine, and methylthioamino acid ester.

3. The bezafibrate salt according to claim 1, characterized in that M + Selected from choline or N-methylnicotinamide.

4. A pharmaceutical composition, characterized in that The bezafibrate salt of claim 1 is used as the main active ingredient.

5. The pharmaceutical composition according to claim 4, characterized in that The drug is administered by injection, and the selected dosage form is sterile powder injection, small water injection or large infusion.

6. The pharmaceutical composition according to claim 4, characterized in that The drug is administered orally, and the selected dosage forms are tablets, coated tablets, capsules, granules, and oral solutions.

7. Use of the bezafibrate salt according to claim 1 in the preparation of a drug for regulating blood lipids.

8. Use of the bezafibrate salt according to claim 1 in the preparation of a medicament for treating hepatobiliary diseases.

9. Use of the pharmaceutical composition according to claim 4 in the preparation of a drug for regulating blood lipids.

10. Use of the pharmaceutical composition according to claim 4 in the preparation of drugs for treating hepatobiliary diseases.

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