Pirfenidone eutectic crystal as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pirfenidone eutectic crystal and a preparation method, a pharmaceutical composition and application thereof. The pirfenidone eutectic provided by the invention is prepared from pirfenidone and a eutectic ligand, and the eutectic ligand is selected from one of pimelic acid, cinnamic acid and saccharin. The pirfenidone eutectic crystal provided by the invention not only can improve the acute pancreatitis relieving effect of pirfenidone, but also can improve the safety of pirfenidone.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to pirfenidone cocrystal and a preparation method thereof, a pharmaceutical composition and use thereof. Background Art

[0002] Pirfenidone is a pyridone compound used clinically to treat idiopathic pulmonary fibrosis. Its specific mechanism of action is still unclear, but it has been reported to inhibit the synthesis and secretion of multiple inflammatory factors, reduce lipid peroxidation, and has anti-inflammatory, antioxidant and anti-fibrotic effects. In addition to clinically approved indications, Shi Xiaoxian et al. reported in the article "Study on the Therapeutic Effect of Pirfenidone on L-arginine-induced Acute Pancreatitis in Mice" that pirfenidone can also relieve L-arginine-induced acute pancreatitis. For the treatment of acute diseases, the absolute safety of drugs is particularly important. It has been reported that pirfenidone has no significant effect on the LD of SW mice. 50 It is 1000mg / kg, and its safety needs to be further improved.

[0003] Currently, there are no reports on the use of pirfenidone and cocrystal ligands to prepare cocrystals to enhance the efficacy of pirfenidone in relieving acute pancreatitis and improve its safety. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a pirfenidone cocrystal and a preparation method thereof, a pharmaceutical composition and use thereof. The pirfenidone cocrystal provided by the present invention can not only improve the effect of pirfenidone in relieving acute pancreatitis, but also improve the safety of pirfenidone.

[0005] The technical solution of the present invention is:

[0006] A pirfenidone cocrystal is prepared from pirfenidone and a cocrystal ligand, wherein the cocrystal ligand is selected from one of pimelic acid, cinnamic acid and saccharin.

[0007] Furthermore, the co-crystal ligand is selected from saccharin.

[0008] Furthermore, the molar ratio of pirfenidone to the cocrystal ligand is 1:(0.5-2).

[0009] Furthermore, the molar ratio of pirfenidone to the cocrystal ligand is 1:1.

[0010] Furthermore, the X-ray powder diffraction pattern of the co-crystal expressed as 2θ±0.02° has characteristic peaks at 10.75°, 13.66°, 15.47°, 17.64°, 17.82°, 18.73°, 20.88°, 21.7°, 22.34°, and 25.43°.

[0011] Furthermore, the pirfenidone cocrystal was prepared by wet grinding.

[0012] Furthermore, the solvent used in preparing the pirfenidone cocrystal by wet grinding is selected from one of acetone, ethanol and N,N-dimethylformamide.

[0013] Furthermore, when the pirfenidone cocrystal is prepared by wet grinding, the mass volume ratio of pirfenidone to solvent is 3: (1.5-2.5) mg / μL.

[0014] Furthermore, when the pirfenidone cocrystal was prepared by wet grinding, the mass volume ratio of pirfenidone to solvent was 3:2 mg / μL.

[0015] Furthermore, when the pirfenidone cocrystal is prepared by wet grinding, the grinding time is 1.5 to 2.5 hours.

[0016] Furthermore, when the pirfenidone cocrystal is prepared by wet grinding, the grinding time is 2 hours.

[0017] Another object of the present invention is to provide a pharmaceutical composition, which comprises the above-mentioned pirfenidone cocrystal.

[0018] Furthermore, the pharmaceutical composition is an oral solid preparation, which is prepared with the above-mentioned pirfenidone cocrystal as an active ingredient and pharmaceutically acceptable additives.

[0019] Furthermore, the oral solid preparation is selected from one of granules, tablets and capsules.

[0020] Another object of the present invention is to provide the use of the above-mentioned pirfenidone cocrystal, the pirfenidone cocrystal prepared by the above-mentioned method for preparing the pirfenidone cocrystal, and the above-mentioned pharmaceutical composition in the preparation of drugs for treating pancreatitis or pulmonary fibrosis.

[0021] Drug cocrystal refers to a new solid phase crystal formed by a fixed stoichiometric ratio of an API and a cocrystal conformer (CCF) under the action of hydrogen bonds, π-π stacking or other non-covalent bonds. In addition to the improvement of physical and chemical properties such as solubility and stability, drug cocrystals may (but not necessarily) bring about improvements in in vivo effects such as pharmacodynamics, pharmacokinetics and toxicology. However, whether a specific API and CCF can form a cocrystal and whether the formed cocrystal can achieve the expected improvement are both very uncertain. There is no technical inspiration in the prior art to improve the effect of pirfenidone in relieving acute pancreatitis and improve its safety through cocrystal technology. The pirfenidone cocrystal prepared by pirfenidone and a cocrystal ligand selected from pimelic acid, cinnamic acid and saccharin of the present invention can not only improve the effect of pirfenidone in relieving acute pancreatitis, but also improve its safety.

[0022] Compared with the prior art, the present invention has the following advantages:

[0023] (1) The experimental results of the effect on L-arginine-induced acute pancreatitis in mice showed that the serum amylase level of the co-crystal prepared by the present invention was statistically significantly lower than that of the pirfenidone group, which indicates that the pirfenidone co-crystal prepared by the present invention can improve the effect of pirfenidone in relieving acute pancreatitis.

[0024] (2) The results of the safety study showed that, under the condition that the oral dose was higher than that of the pirfenidone group, the number of animals that died within one week in the cocrystal group that was orally administered with the pirfenidone cocrystal prepared by the present invention was still significantly lower than that in the pirfenidone group. This indicates that the safety of the pirfenidone cocrystal prepared by the present invention is higher than that of pirfenidone. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 XRPD pattern of cocrystal 3 prepared in the present invention;

[0026] Figure 2 XRPD comparison diagram of cocrystal 3, API after grinding, and dextrin after grinding;

[0027] Figure 3 This is the DSC and TGA overlay of cocrystal 3 prepared in the present invention;

[0028] Figure 4 This is an image of cocrystal 3 prepared in the present invention under a polarizing microscope. DETAILED DESCRIPTION

[0029] The present invention is further illustrated below through the description of specific implementation modes, but this is not a limitation of the present invention. Those skilled in the art can make various modifications or improvements based on the basic idea of ​​the present invention, but as long as they do not deviate from the basic idea of ​​the present invention, they are all within the scope of the present invention.

[0030] Example 1 Cocrystal screening test

[0031] 1.1 Materials

[0032] 1.1.1 Reagents

[0033] The reagents used in this embodiment include:

[0034] API: Pirfenidone

[0035] Cocrystal ligands: pimelic acid, sebacic acid, lauric acid, myristic acid, cinnamic acid, palmitic acid, stearic acid, pamoic acid, saccharin and fumaric acid, etc.

[0036] Solvents: acetone, ethanol, N,N-dimethylformamide.

[0037] The above reagents are all highly commercially available.

[0038] 1.1.2 Equipment and Instruments

[0039] The equipment and instruments used in this embodiment are all commonly used equipment and instruments for wet grinding, and are also equipment and instruments that can be equipped by ordinary pharmaceutical research and development institutions.

[0040] 1.2 Methods

[0041] a) weighing 30 mg of pirfenidone sample and an equal molar ratio of ligand into a mortar;

[0042] b) adding 20 μL of solvent;

[0043] c) After grinding for 2 h, the solid samples were collected and dried to constant weight, and then the melting point (melting range) was measured. The results are shown in Table 1.

[0044] 1.3 Results

[0045] The results are shown in Table 1.

[0046] Table 1 Cocrystal screening test results

[0047] API Cocrystal ligand Solvents Product melting range (℃) Eutectic number Pirfenidone Lauric acid acetone >2 Pirfenidone Sebacic acid DMF >2 Pirfenidone Pimelic acid Ethanol <2 Eutectic 1 Pirfenidone Myristic acid acetone >2 Pirfenidone Cinnamic acid DMF <2 Eutectic 2 Pirfenidone Palmitic acid Ethanol >2 Pirfenidone saccharin acetone <2 Eutectic 3 Pirfenidone Stearic acid DMF >2 Pirfenidone Pamoic acid Ethanol >2 Pirfenidone Fumaric Acid acetone >2

[0048] 30 mg of pirfenidone sample was weighed, 20 μL of ethanol was added, and after grinding for 2 h, the obtained solid sample was collected and dried to constant weight, and then the melting point A1 was measured.

[0049] 30 mg of pirfenidone sample was weighed, 20 μL of DMF was added, and after grinding for 2 h, the obtained solid sample was collected and dried to constant weight, and then the melting point B1 was measured.

[0050] Weigh 30 mg of pirfenidone sample, add 20 μL of acetone, grind for 2 h, collect the obtained solid sample, dry to constant weight, and measure the melting point C1.

[0051] Weigh an amount of pimelic acid equimolar to 30 mg of pirfenidone, add 20 μL of ethanol, grind for 2 h, collect the resulting solid sample, dry it to constant weight, and measure the melting point A2.

[0052] Cinnamic acid in an amount equal to 30 mg of pirfenidone was weighed, 20 μL of DMF was added, and after grinding for 2 h, the resulting solid sample was collected and dried to constant weight, and the melting point B2 was measured.

[0053] Saccharin in an amount equal to 30 mg of pirfenidone was weighed, 20 μL of acetone was added, and after grinding for 2 h, the obtained solid sample was collected and dried to constant weight, and then the melting point C2 was measured.

[0054] After calculation, |A1-A2|, |B1-B2| and |C1-C2| were all >5°C, indicating that the corresponding API-cocrystal ligand-solvent combination formed a single compound.

[0055] The elemental analysis results showed that the molar ratio of the API to the cocrystal ligand in cocrystal 1, cocrystal 2 and cocrystal 3 was 1:1, and all of them were non-solvates.

[0056] The results in Table 1 further confirm that whether the same API can form a cocrystal with a cocrystal ligand is highly unpredictable.

[0057] Example 2 Screening of the superior cocrystal 1 - Effect on L-arginine-induced acute pancreatitis in mice

[0058] Basically, the method disclosed in the article "Study on the Therapeutic Effect of Pirfenidone on L-arginine-induced Acute Pancreatitis in Mice" by Shi Xiaoxian et al. was used to investigate the effects of cocrystal 1, cocrystal 2, cocrystal 3 in Table 1 and pirfenidone raw material on L-arginine-induced acute pancreatitis in mice, among which the results of serum amylase level determination in each group are shown in Table 2.

[0059] Table 2 Serum amylase level determination results of each group

[0060]

[0061] Note:

[0062] a All are calculated based on pirfenidone.

[0063] The results of the two-sided equal variance t test (Table 2) showed that:

[0064] The serum amylase level in the model group was statistically significantly higher than that in the blank control group;

[0065] The serum amylase levels in the pirfenidone group, cocrystal group 1, cocrystal group 2, and cocrystal group 3 were statistically significantly lower than those in the model group;

[0066] The serum amylase levels in cocrystal group 1 and cocrystal group 3 were statistically significantly lower than that in the pirfenidone group.

[0067] Example 3 Advantageous Crystal Form Screening 2 - Safety Study

[0068] C57BL / 6 mice, weighing 20-25 g and aged 4-6 weeks, were randomly divided into 3 groups, with 5 males and 5 females in each group. The drugs and doses shown in Table 3 were administered by single oral gavage, and the animals were then allowed to eat, drink and move freely. The number of animals that died within 1 week was calculated, and the results are shown in Table 3.

[0069] Table 3 Safety study results

[0070] Group N dose Number of dead animals Pirfenidone group 10 1500mg / kg 8 Eutectic-1 Group 10 3000mg / kg 6 Eutectic-Group 3 10 3000mg / kg 3

[0071] As shown in Table 3, the LD of eutectic 3 50 >3000mg / kg, significantly higher than pirfenidone.

[0072] Example 4 Further structural confirmation of cocrystal 3

[0073] 4.1 X-ray powder diffraction (XRPD)

[0074] The relevant parameters of the XRPD instrument are shown in Table 4:

[0075] Table 4 XRPD instrument related parameters

[0076] instrument Malvern Panalytical Aeris X-ray wavelength Cu, Kα:1.5419 X-ray pipeline setup 40kV,15mA Scan Mode 1D Scanning range (2θ) 3.5o-40o Scan step length (2θ) 0.0027166o Scan rate (2θ) 10o / min

[0077] The XRPD pattern of the cocrystal 3 prepared by the present invention is as follows Figure 1 As shown, the XRPD comparison of cocrystal 3, API after grinding, and dextrin after grinding is shown in Figure 2 shown. Figure 1 , Figure 2 R70324044-Sac represents eutectic 3. Figure 2 API stands for pirfenidone. Figure 1 The positions and relative intensities of the Top 10 absorption peaks are shown in Table 5.

[0078] Depend on Figure 1 From the results in Table 5, it can be seen that the X-ray powder diffraction pattern of the cocrystal 3 prepared by the present invention expressed as 2θ±0.02° has characteristic peaks at 10.75°, 13.66°, 15.47°, 17.64°, 17.82°, 18.73°, 20.88°, 21.7°, 22.34°, and 25.43°. Figure 2 It can be seen that cocrystal-3 exhibits good crystallinity.

[0079] Table 5 Figure 1 Top 10 absorption peak positions and relative intensities (%)

[0080] Position (2θ±0.02°) Relative strength (%) 10.75 100 13.66 35.91 15.47 21.78 17.64 16.96 17.82 15.54 18.73 30.56 20.88 25.57 21.7 29 22.34 18.47 25.43 51.89

[0081] 4.2 Thermogravimetric analysis (TGA)

[0082] The relevant parameters of the TGA instrument are shown in Table 6:

[0083] Table 6 TGA instrument related parameters

[0084] Experimental Instruments TA,TGA500 How to use Linear temperature rise Temperature range RT-300.00℃ Heating rate 10℃ / min Purge gas <![CDATA[N2(>99.999%)]]> Sample tray type Aluminum sample pan, open without cover

[0085] 4.3 Differential Scanning Calorimetry (DSC)

[0086] The relevant parameters of the DSC instrument are shown in Table 7:

[0087] Table 7 Related parameters of DSC instrument

[0088] Experimental Instruments TA,DSC250 How to use Linear temperature rise Temperature range RT-300.00℃ Heating rate 10℃ / min Purge gas <![CDATA[N2(>99.999%)]]> Sample tray type Aluminum sample pan with lid, perforated top

[0089] The DSC and TGA superposition diagrams of the cocrystal 3 prepared by the present invention are as follows: Figure 3 shown. Figure 3 R70324044-SAC represents eutectic 3. Figure 3 The TGA test results show that the material weight loss (Weight Percent Loss) before melting is 0.483%, and the mass loss (Weight Loss) is 0.013mg. Figure 3 The DSC test results show that the material starts to melt at a temperature of 119.32°C, the enthalpy (normalized) is 85.178 J / g, and the peak temperature is 120.45°C.

[0090] 4.3 Polarized light microscopy (PLM)

[0091] The samples to be tested were spread on a glass slide with methyl silicone oil and observed by PLM.

[0092] The image of the eutectic 3 prepared by the present invention under a polarizing microscope is as follows: Figure 4 As shown. Figure 4 It can be seen that under the test condition that the magnification of the polarizing microscope is 10*2.5 times, the eutectic 3 prepared by the present invention is small, irregular crystal particles.

[0093] Example 5 Preparation Example

[0094] (1) Capsules

[0095] The capsule formulation containing cocrystal 1 (100 capsules) is shown in Table 8.

[0096] Table 8 Capsule formulation containing cocrystal 1 (100 capsules)

[0097] Eutectic 1 3g Microcrystalline Cellulose 20g Glyceryl Monostearate 0.8g

[0098] Preparation method of capsules: pass cocrystal 1, microcrystalline cellulose and glyceryl monostearate through an 80-mesh sieve for later use. Mix the prescribed amount of cocrystal 1, microcrystalline cellulose and glyceryl monostearate and fill the capsules.

[0099] (2) Tablets

[0100] The tablet formulation containing cocrystal 3 (100 tablets) is shown in Table 9.

[0101] Table 9 Tablet formula containing cocrystal 3 (based on 100 tablets)

[0102] Eutectic 3 3.5g Sorbitol 18g Glyceryl Monopalmitate 0.8g

[0103] Preparation method of tablets: pass cocrystal 3, sorbitol and monopalmitin through an 80-mesh sieve for later use. Mix the prescribed amount of cocrystal 3, sorbitol and monopalmitin and compress into tablets.

Claims

1. A pirfenidone cocrystal, characterized in that The co-crystal is prepared from pirfenidone and a co-crystal ligand, and the co-crystal ligand is selected from one of pimelic acid, cinnamic acid and saccharin.

2. The pirfenidone cocrystal according to claim 1, characterized in that The molar ratio of pirfenidone to the cocrystal ligand is 1:(0.5-2).

3. The pirfenidone cocrystal according to claim 1, characterized in that The X-ray powder diffraction pattern of the co-crystal expressed as 2θ±0.02° has characteristic peaks at 10.75°, 13.66°, 15.47°, 17.64°, 17.82°, 18.73°, 20.88°, 21.7°, 22.34°, and 25.43°.

4. The method for preparing the pirfenidone cocrystal according to any one of claims 1 to 3, characterized in that: Pirfenidone cocrystals were prepared by wet grinding.

5. The method for preparing the pirfenidone cocrystal according to claim 4, characterized in that: The solvent used in preparing pirfenidone cocrystal by wet grinding is selected from one of acetone, ethanol and N,N-dimethylformamide.

6. The method for preparing the pirfenidone cocrystal according to claim 4, characterized in that: When the pirfenidone cocrystal is prepared by wet grinding, the mass volume ratio of pirfenidone to solvent is 3: (1.5-2.5) mg / μL.

7. The method for preparing the pirfenidone cocrystal according to claim 4, characterized in that: When the pirfenidone cocrystal is prepared by wet grinding, the grinding time is 1.5 to 2.5 hours.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the pirfenidone co-crystal according to any one of claims 1 to 3.

9. The pharmaceutical composition according to claim 8, characterized in that The pharmaceutical composition is an oral solid preparation, which is prepared using the pirfenidone cocrystal described in any one of claims 1 to 3 as an active ingredient and a pharmaceutically acceptable additive.

10. Use of the pirfenidone co-crystal according to any one of claims 1 to 3, the pirfenidone co-crystal prepared by the method for preparing the pirfenidone co-crystal according to any one of claims 4 to 7, or the pharmaceutical composition according to claim 8 or claim 9 in the preparation of a medicament for treating pancreatitis or pulmonary fibrosis.

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