Nintedanib derivative as well as preparation method and application thereof

Abstract

The invention relates to a crystal form of a novel nintedanib derivative as shown in a formula (I), a preparation method of the crystal form, and medical application of a medicinal preparation containing the crystal form. The invention further provides application of the compound shown in the formula (I) in preparation of drugs for treating systemic sclerosis related pulmonary interstitial diseases, idiopathic pulmonary fibrosis and non-small cell lung cancer. The compound has the advantages of being remarkable in stability and long-acting.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a nintedanib derivative, its preparation method, and its pharmaceutical formulation and medicinal uses. Background Technology

[0002] Currently, only two new compound drugs for the treatment of pulmonary fibrosis have been approved for marketing worldwide: pirfenidone and nintedanib. Due to the significant adverse reactions (AEs) of these two drugs (photosensitivity rash (51.7%), loss of appetite (23.0%), stomach upset (14%), nausea (12.1%), etc.), patients often cannot tolerate them. Therefore, the actual therapeutic dose in clinical practice does not reach the daily dose used in clinical studies. A post-marketing surveillance report for pirfenidone in Japan showed that 61.8% of patients frequently received doses below 1200 mg / day during treatment. This situation is also observed in Chinese patients, with clinical treatment generally maintained at a dose of 1200 mg / day, and reaching a dose of 1800 mg / day being extremely difficult. A retrospective RWD clinical study conducted in China showed that pulmonary fibrosis patients often drop out due to intolerance to adverse drug reactions. The dropout rates for nintedanib and pirfenidone were over 61.22% and 32.68%, respectively (Dai et al., 2024), which are very high.

[0003] Nintedanib is a tyrosine kinase inhibitor that inhibits vascular endothelial growth factor and other pro-fibrotic mediators, such as platelet-derived growth factor, transforming growth factor, and fibroblast growth factor, and is active against fibroblasts. Without treatment, the life expectancy of patients is close to 3-5 years; with nintedanib, it is known to extend life to 7-11 years (Hilberg et al., 2008). Nintedanib has low oral bioavailability (<5%) and a high initial metabolic rate (Wind et al., 2019). The pharmacokinetics of nintedanib are well-established, showing rapid absorption after oral administration, high protein binding capacity, wide tissue distribution, metabolism via multiple enzymes, and primary excretion in feces. The recommended daily dose range for nintedanib is 100-150 mg. However, it can cause diarrhea, nausea, vomiting, and gastrointestinal side effects, and carries a risk associated with cardiovascular and bleeding events (Rivera-Ortega et al., 2018).

[0004] To enhance its bioavailability and therapeutic potential, several novel nintedanib formulations have been developed. Cyclodextrin complexes of nintedanib are designed to reduce glycoprotein excretion and improve its transport across the intestinal membrane (Vaidya et al., 2019). Similarly, liposomes (Kala and Chinni, 2022), polymeric nanoparticles (Liu et al., 2018), solid lipid nanocarriers (Zhu et al., 2020), and self-microemulsification delivery (Liu et al., 2019) have been investigated to improve drug release, bioavailability, and therapeutic efficacy. Furthermore, studies have investigated nanocrystals that improve mucosal adhesion and absorption (Zhu et al., 2022), magnetic nanospheres with therapeutic applications (Karade et al., 2021), and dry powder inhalers, demonstrating experimental reductions in pulmonary collagen deposition (Surber et al., 2020).

[0005] Optimizing drug bioavailability has many potential benefits. Reducing dosing frequency is generally considered desirable for patient convenience and improved adherence. By extending the drug release time, the duration of action per dose is expected to be longer. This then leads to an overall improvement in dosing parameters, such as taking the drug once daily instead of four times daily or once weekly, or even less when daily dosing was previously required. Many drugs are currently administered once daily. However, not all of these drugs have pharmacokinetic properties suitable for an exact 24-hour dosing interval. To date, no long-acting injectable drugs for the treatment of idiopathic pulmonary fibrosis have been approved on the market. Furthermore, successful optimization is required to determine key parameters associated with the development of various clinical drug candidates or molecules, such as selecting the appropriate dose, dosage form, route of administration, dosing regimen, and effects on animals and humans. Therefore, there is an unmet need for an effective drug delivery system for nintedanib.

[0006] The bioavailability of nintedanib can be improved via the injection route; more specifically, long-acting (sustained-release or extended-release) injectable formulations may provide better bioavailability after intramuscular or subcutaneous administration than the oral route. The anticipated increase in bioavailability of injectable formulations can provide therapeutic plasma concentration levels, with doses administered intramuscularly or subcutaneously once daily, every three days, weekly, every two weeks, every three weeks, monthly, every two months, every three months, or every six months. The total injectable dose can be significantly lower than the daily oral dose required for the same period, thereby reducing toxicity and improving patient compliance. Therefore, given the advantages associated with long-acting formulations, the need to develop long-acting parenteral formulations of nintedanib remains unmet, which would provide higher compliance while maintaining therapeutic levels of the drug in the patient system for extended periods (days, weeks, months, or even years) for the treatment of systemic sclerosis-related interstitial lung disease; idiopathic pulmonary fibrosis; non-small cell lung cancer; or other related diseases described herein.

[0007] Non-oral administration routes (e.g., parenteral routes) have been explored for use in other classes of drugs. However, to date, there is no extended-release injectable prodrug formulation of nintedanib.

[0008] Prior art regarding nintedanib derivatives includes: WO2022126441, CN117777003, CN103848814, Chemical Communications (Cambridge, United Kingdom) (2022), 58(8), 1199-1202, and WO2022012492. To overcome the aforementioned deficiencies of existing nintedanib and its derivatives, this invention provides a long-acting nintedanib derivative, its preparation method, and its pharmaceutical application, thereby enhancing the pharmaceutical activity and practicality of existing nintedanib and its derivatives. Summary of the Invention

[0009] This invention modifies the structure of nintedanib to prepare a prodrug with long-acting properties. This type of drug is formulated into a preparation suitable for intramuscular, subcutaneous, or intravenous injection. After intramuscular, subcutaneous, or intravenous injection, a drug reservoir is formed in the body. The drug is slowly, continuously, and stably released from the reservoir and converted into nintedanib, thereby exerting a long-acting effect. This invention is achieved using the following technical solution: The purpose of this invention is to overcome the shortcomings of the prior art and provide a nintedanib derivative or its solvate or a pharmaceutically acceptable salt thereof as shown in formula (I): .

[0010] The X-ray powder diffraction pattern of the compound represented by Formula I, expressed as a diffraction angle of 2θ±0.2°, is located at 6.52±0.2°, 7.88±0.2°, 8.70±0.2°, 10.12±0.2°, 11.68±0.2°, 12.60±0.2°, 13.14±0.2°, 15.60±0.2°, 15.88±0.2°, 16.34±0.2°, 17.46±0.2°, 17.80±0.2°, 18.80±0.2°, 19.42±0.2°, 19.74±0.2°, 20.34±0.2°, 2 Characteristic peaks are observed at 1.72±0.2°, 22.20±0.2°, 23.18±0.2°, 23.94±0.2°, 24.82±0.2°, 25.72±0.2°, 26.52±0.2°, 26.80±0.2°, 26.90±0.2°, 26.96±0.2°, 27.70±0.2°, 28.06±0.2°, 28.60±0.2°, 29.04±0.2°, 29.54±0.2°, 29.84±0.2°, 29.96±0.2°, and 30.78±0.2°. Its X-ray powder diffraction pattern is as follows: Figure 2 As shown.

[0011] Another object of the present invention is to provide a method for preparing nintedanib derivatives of Formula I or their solvates or pharmaceutically acceptable salts thereof, comprising the following steps: Step 1: Nydanib and chloromethylene laurate are reacted with a catalyst and solvent to prepare crude product (I); Step 2: Prepare pharmaceutical-grade raw material of formula (I) from crude product of formula (I) in a crystallization solvent, as shown in the following reaction formula:

[0012] In step 1, the reaction solvent is selected from one or more combinations of N,N-dimethylformamide, N,N-dimethylacetamide, DMSO, acetonitrile, and tetrahydrofuran, preferably acetonitrile; the catalyst is selected from cesium carbonate and pyridine DMAP, preferably cesium carbonate; the crystallization solvent in step 2 is selected from one or more combinations of n-pentane, n-hexane, n-heptane, cyclohexane, petroleum ether, toluene, xylene, dimethyl ether, diethyl ether, isopropyl ether, tetrahydrofuran, methyl acetate, ethyl acetate, dichloromethane, dichloroethane, chloroform, anhydrous ethanol, and anhydrous methanol, preferably anhydrous methanol and a combination of ethyl acetate / n-heptane, and the crystallization temperature is 0 to -20°C, preferably 0 to -5°C.

[0013] As a preparation method, the inventors also studied the following general method, with the reaction formula as follows:

[0014] As a result, the product could not be purified in step 1, making it impossible to prepare the pharmaceutical standard formula (Ⅰ).

[0015] In optimizing the process, a series of crystallization methods were screened to prepare the crystal form of the compound of the present invention, and the results are as follows:

[0016] Based on the above screening, the combination of anhydrous methanol and ethyl acetate / n-heptane as solvents for the crystallization process we provide is preferred, with the ethyl acetate / n-heptane (V / V) ratio preferably being 1:4 to 6; and the crystallization temperature preferably being -20 to 5°C.

[0017] Further research on the properties of the crystals revealed that the crystals obtained in this invention have many advantages, such as good chemical stability, few impurities under long-term storage conditions, and good hygroscopicity, which are beneficial for the further development of corresponding drugs.

[0018] In some embodiments, the purity and structure of the product prepared by the present invention were confirmed by chromatographic spectroscopy analysis, and the chromatographic analysis conditions are as follows: Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (size: 150×4.6mm; 3μm). Mobile phase A was 5mmol / L potassium dihydrogen phosphate solution (containing 0.1% triethylamine, pH adjusted to 6.0 with phosphoric acid)-tetrahydrofuran (90:10), and mobile phase B was acetonitrile-tetrahydrofuran (90:10). Gradient elution was performed according to the table below. Detection wavelength was 287nm. Column temperature was 35℃. Flow rate was 1.0ml per minute. Injection volume was 10μl.

[0019]

[0020] The assay involves precisely injecting the test solution into the liquid chromatograph and recording the chromatogram. Typical chromatograms and spectra are shown in the attached figure.

[0021] Another object of the present invention is a pharmaceutical composition comprising a nintedanib derivative or a solvate thereof as shown in Formula I and a pharmaceutically acceptable carrier. It is formulated for intramuscular, intradermal, or subcutaneous injection. The injectable formulation is prepared from the active ingredient of Formula (I) and related pharmaceutical excipients according to pharmaceutically acceptable methods, including pharmaceutically necessary excipients such as suspending agents, antioxidants, stabilizers, wetting agents, and preservatives. The injectable solution of the present invention can be an aqueous suspension, an oil-based injection, or a lyophilized solution, and is used in the preparation of drugs for the treatment of systemic sclerosis-related interstitial lung disease, idiopathic pulmonary fibrosis, and non-small cell lung cancer.

[0022] The key points of this invention are: 1) The crystal form of the derivative of the present invention has good stability and suitable solubility, and good long-term stability; 2) The crystal form of the derivative of this invention is obtained by cooling crystallization, which is simple, energy-efficient, suitable for industrialization, and has a stable crystal form, which is conducive to further development; 3) The derivatives of the present invention have superior stability and pharmacokinetic advantages compared with similar acetates, hexanoates, decanoates, hexadecyl esters and eicosyl esters, while also having the advantage of low toxicity. Attached Figure Description

[0023] Figure 1 This is the structure of the compound of formula I.

[0024] Figure 2 The XRD pattern is shown for compound I.

[0025] Figure 3 HPLC for compound of formula I.

[0026] Figure 4 This is the hydrogen spectrum of compound I.

[0027] Figure 5 This is the carbon spectrum of compound I.

[0028] Figure 6 LC-MS of compound of formula I.

[0029] Figure 7 The DSC / TG values ​​are for compounds of formula I.

[0030] Figure 8 IR for compound of formula I.

[0031] Figure 9 These are the results of pharmacokinetic studies in rats. Detailed Implementation

[0032] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. The following embodiments can further describe the present invention; however, these embodiments should not be construed as limiting the scope of the present invention.

[0033] Example 1: Preparation of nintedanib lauryl ester.

[0034]

[0035] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0–10 °C, and chloromethylene laurate (3.0 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. After adding 250 mL of dichloromethane and stirring to disperse the mixture, 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH = 15:1) to obtain 5.79 g of reddish-brown oily substance. This was dissolved in 60 mL of anhydrous methanol by heating, and then refrigerated overnight to allow crystallization, yielding 4.53 g of pale yellow crystalline solid, which was the target product. Samples were sent for analysis of 1H NMR, 1C NMR, mass spectrometry, infrared spectroscopy, DSC, and XRD. The results are shown below: 1H-NMR (400MHz, d-DMSO): δ11.9769 (s, 1H); 7.7216~7.4938 (m, 6H); 7.315 4(dd,1H);7.1690(d,2H);6.9668(d,2H);5.8594(s,2H);5.8486(d,1H);3 .7998(s,3H);3.0816(s,3H);2.7111(s,2H);2.3371(t,2H);2.2070(m,8H ); 2.1276 (s, 3H); 1.5129 (m, 2H); 1.2569~1.1683 (m, 16H); 0.8474 (t, 3H).

[0036] 13C-NMR (101MHz, d-DMSO): δ172.97, 168.39, 166.70, 160.06, 141.21, 135.85, 132.20, 131.15, 129.99, 128.91, 124.74, 123.4 2, 117.83, 109.68, 63.07, 55.04, 52.76, 52.35, 46.16, 33.79, 31.76, 29.42, 29.31, 29.14, 29.12, 28.76, 24.84, 22.55, 14.41.

[0037] Mass spectrometry: 750.9 = MH.

[0038] Infrared spectrum: 3446.41, 2923.47, 2851.78, 2798.26, 1742.87, 1715.44, 1675.52, 1653.51, 1621.72, 1596.43, 1571.23, 1540.48, 1516.25, 1474.07, 1456.96, 1436.59, 1375.85, 1350.84, 1318 .37, 1287.26, 1253.37, 1222.78, 1202.82, 1147.09, 1113.74, 1091.91, 1064.38, 1012.18, 980.95, 926.61, 847.84, 830.20, 770.31, 734.78, 717.75, 686.56, 625.50, 570.07, 532.62.

[0039] DSC-TGA: This product does not contain water of crystallization or crystallization solvent. Its melting point is approximately 124.75℃, and it begins to decompose at around 280℃.

[0040] XRD: This product is a crystalline powder with characteristic 2θ angles of: 6.52, 7.88, 8.70, 10.12, 11.68, 12.60, 13.14, 15.60, 15.88, 16.34, 17.46, 17.80, 18.80, 19.42, 19.74, 20.34, 21.72, 22.20, 23.18, 23.94, 24.82, 25.72, 26.52, 26.80, 26.90, 26.96, 27.70, 28.06, 28.60, 29.04, 29.54, 29.84, 29.96, and 30.78.

[0041] Example 2: Preparation of nintedanib lauryl ester.

[0042] Nintanib (0.54 g, 1 mmol), dichloromethane (20 mL), and cesium carbonate (0.36 g, 1.1 mmol) were added to a 100 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0–10 °C, and chloromethylene laurate (0.3 g, 1.2 mmol) was added. The mixture was stirred at 36–40 °C overnight. At room temperature, 100 mL of dichloromethane and 100 mL of water were added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (20 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=15:1) to obtain 0.47 g of reddish-brown oily substance. 5 mL of anhydrous methanol was added and heated to dissolve the substance. The mixture was then refrigerated overnight to allow crystallization, yielding 0.41 g of pale yellow crystalline solid, which was the target product.

[0043] Example 3: Preparation of nintedanib lauryl ester.

[0044] Nintedanib (5.4 g, 10 mmol), acetone (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0–10 °C, and chloromethylene laurate (3.0 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. After adding 250 mL of dichloromethane and stirring to disperse the mixture, 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=15:1) to obtain 5.32 g of reddish-brown oily substance. 55 mL of anhydrous methanol was added and heated to dissolve the oil. The solution was then placed in a refrigerator and allowed to crystallize overnight to obtain 4.13 g of pale yellow crystalline solid, which was the target product.

[0045] Example 4: Preparation of nintedanib lauryl ester.

[0046] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and alkalized by stirring at 40 °C for 30 min. The temperature was lowered to 0–10 °C, and chloromethylene laurate (3.8 g, 15 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. After adding 250 mL of dichloromethane and stirring to disperse, 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=15:1) to obtain 5.94 g of reddish-brown oily substance. Ethyl acetate / n-heptane (2V / 10V) was added for recrystallization, and the mixture was placed in a refrigerator for overnight crystallization to obtain 4.82 g of pale yellow crystalline solid, which was the target product.

[0047] Example 5: Preparation of nintedanib lauryl ester.

[0048] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0–10 °C, and chloromethylene laurate (5.0 g, 20 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. After adding 250 mL of dichloromethane and stirring to disperse the mixture, 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=15:1) to obtain 5.80 g of reddish-brown oily substance. Ethyl acetate / n-heptane (2V / 10V) was added for recrystallization, and the mixture was placed in a refrigerator for overnight crystallization to obtain 4.64 g of pale yellow crystalline solid, which was the target product.

[0049] Example 6: Preparation of nintedanib lauryl ester.

[0050] Nintanib (5.4 g, 10 mmol), DMF (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0–10 °C, and chloromethylene laurate (3.0 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. After adding 250 mL of dichloromethane and stirring to disperse the mixture, it was washed twice with 100 mL × 2 of water. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=15:1) to obtain 1.56 g of reddish-brown oily substance. 20 mL of anhydrous methanol was added for recrystallization, and the mixture was refrigerated overnight to obtain 1.13 g of pale yellow crystalline solid, which was the target product.

[0051] Comparative Example 1: Preparation of nintedanib acetate.

[0052]

[0053] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl chloroacetate (1.3 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=8:1) to obtain 4.30 g of pale yellow solid. Ethyl acetate / n-heptane (2V / 10V) was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 3.44 g of pale yellow crystalline solid, which was the target product.

[0054] Comparative Example 2: Preparation of nintedanib acetate.

[0055]

[0056] Nintedanib (5.4 g, 10 mmol), dichloromethane (100 mL), and triethylamine (1.2 g, 12 mmol) were added to a 250 mL three-necked flask. The mixture was cooled to 0–10 °C, and methyl chloroacetate (1.3 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. 100 mL of water was added and the mixture was stirred to separate the layers. The lower organic phase was washed twice with a saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=8:1) to obtain 1.94 g of pale yellow solid. Ethyl acetate / n-heptane (2V / 10V) was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 1.44 g of pale yellow crystalline solid, which was the target product.

[0057] Comparative Example 3: Preparation of nintedanibhexanoate.

[0058]

[0059] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl hexanoate (2.0 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=10:1) to obtain 4.50 g of pale yellow solid. 45 mL of methanol was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 3.86 g of pale yellow crystalline solid, which was the target product.

[0060] Comparative Example 4: Preparation of nintedanibutyrate.

[0061] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl hexanoate (2.5 g, 15 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=10:1) to obtain 4.34 g of pale yellow solid. 45 mL of methanol was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 3.64 g of pale yellow crystalline solid, which was the target product.

[0062] Comparative Example 5: Preparation of nintedanib-decanoate.

[0063]

[0064] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl decanoate (2.6 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=12:1) to obtain 4.48 g of pale yellow solid. 45 mL of methanol was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 3.52 g of pale yellow crystalline solid, which was the target product.

[0065] Comparative Example 6: Preparation of nintedanib-decanoate.

[0066] Nintedanib (5.4 g, 10 mmol), acetone (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl decanoate (2.6 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH = 12:1) to obtain 4.25 g of pale yellow solid. 45 mL of methanol was added for recrystallization, and the solid was placed in a refrigerator for overnight crystallization to obtain 3.14 g of pale yellow crystalline solid, which was the target product.

[0067] Comparative Example 7: Preparation of nintedanib hexadecanoate.

[0068]

[0069] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl palmitate (3.7 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15–20 °C. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=18:1) to obtain 6.1 g of pale yellow oily substance. 60 mL of methanol was added for recrystallization, and the mixture was placed in a refrigerator for overnight crystallization. No crystallization occurred. The product was concentrated to obtain 5.6 g of pale yellow oily substance, which was the target product.

[0070] Comparative Example 8: Preparation of nintedanib-hexadecanoate.

[0071] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was then lowered to 0-10 °C, and methyl palmitate (3.7 g, 12 mmol) was added. The mixture was stirred at 40 °C for 6 hours to react. The reaction solution was then concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=18:1) to obtain 5.8 g of pale yellow oily substance. 60 mL of methanol was added for recrystallization, and the mixture was placed in a refrigerator for overnight crystallization. No crystallization occurred. The product was concentrated to obtain 5.2 g of pale yellow oily substance, which was the target product.

[0072] Comparative Example 9: Preparation of nintedanib-eicosinate.

[0073]

[0074] Nintanib (5.4 g, 10 mmol), acetonitrile (100 mL), and cesium carbonate (3.6 g, 11 mmol) were added to a 250 mL three-necked flask and stirred at 40 °C for 30 min to alkalize. The temperature was lowered to 0–10 °C, and methyl chloroeicosate (4.3 g, 12 mmol) was added. The mixture was stirred at 15–20 °C overnight. The reaction solution was concentrated under reduced pressure at 45 °C. 250 mL of dichloromethane was added and stirred to disperse the mixture. 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=20:1) to obtain 6.3g of pale yellow oily substance. 65mL of methanol was added for recrystallization, and the mixture was refrigerated overnight for crystallization. No crystallization occurred. The product was concentrated to obtain 5.7g of pale yellow oily substance, which was the target product.

[0075] Comparative Example 10: Preparation of nintedanib-eicosinate.

[0076] Nintanib (5.4 g, 10 mmol), DMF (100 mL), and 60% sodium hydroxide (0.48 g, 12 mmol) were added to a 250 mL three-necked flask and stirred at room temperature for 30 min to alkalize. The temperature was then lowered to 0-10 °C, and methyl chloroeicosinate (4.3 g, 12 mmol) was added. The mixture was stirred and reacted overnight at 15-20 °C. After adding 250 mL of dichloromethane and stirring to disperse the mixture, 100 mL of water was added and stirred to separate the layers. The lower organic phase was washed twice with saturated sodium bicarbonate aqueous solution (150 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain an oily substance. The oily substance was purified by column chromatography (DCM:MeOH=20:1) to obtain 5.3g of pale yellow oily substance. 65mL of methanol was added for recrystallization, and the mixture was placed in a refrigerator for overnight crystallization. No crystallization occurred. The product was concentrated to obtain 4.8g of pale yellow oily substance, which was the target product.

[0077] Test Example 1: Physicochemical Properties.

[0078]

[0079] Solubility of this invention:

[0080] Nydanib solubility:

[0081] Conclusion: The solubility of this invention is orders of magnitude higher than that of nintedanib.

[0082] Test Example 2: Crystal Stability Test.

[0083] 1. Grinding test: Weigh 1g of the nidanib derivative crystal of this invention into an agate mortar and grind thoroughly for half an hour. Collect the sample and weigh 0.82g. X-ray powder diffraction confirmed that the product is the original crystal of this invention, and the crystal form did not change under these conditions.

[0084] 2. Influencing Factors Experiment The crystals of the nintedanib derivative of this invention were packaged in pharmaceutical low-density polyethylene bags and pharmaceutical composite films of polyester / aluminum / polyethylene, and subjected to tests under various influencing conditions, including light exposure (total fluorescent lamp illuminance not less than 1.2 × 10⁶ Lux·hr and near-ultraviolet lamp energy not less than 200 W·hr / m²), high temperature (60°C), and high humidity (25°C, RH: 90% ± 5, 30 days of high humidity). The results are as follows:

[0085] The data in the table show that the crystal form of the nintedanib lauroyl ester derivative of the present invention can maintain good stability under high temperature, high humidity and light conditions, indicating that the crystal form provided by the present invention has good stability and is beneficial to the storage of the active pharmaceutical ingredient.

[0086] Test Example 3: Stability Test.

[0087] The stability of compounds is generally unpredictable. Since the compounds of this invention may be used as active pharmaceutical ingredients (APIs) in the preparation of long-acting injectable solutions, storage is crucial. Impurities may be introduced or generated during storage, affecting the use of the API; therefore, the stability of the API of this invention is investigated. Following the ICH guideline "Stability Testing of New APIs and Formulations," influencing factor tests were conducted.

[0088] 1. Test conditions: The high humidity test of this product was conducted at 25℃±2℃, RH: 75%±5%, and high temperature (60℃) with the inner and outer packaging removed for 30 days. The light exposure (total illuminance not less than 1.2×106Lux·hr) was also conducted with the inner and outer packaging removed.

[0089] 2. Sampling and testing: High temperature samples were taken at 5, 10 and 30 days; high humidity samples were taken at 5 and 10 days; and light conditions samples were taken at 5 and 11 days.

[0090] 3. Observation results: See the table below.

[0091] Conclusion: As shown in the table, the active pharmaceutical ingredient of the present invention is relatively stable under the relevant conditions and no obvious impurities are generated, while similar compounds show obvious instability at high temperatures.

[0092] Test Example 4: Experimental study on in vitro cytotoxicity of normal human hepatocytes.

[0093] 1. Experimental Materials 1.1 Cells: LO2 cells, a human hepatitis cell line 1.2 Drug: Compound of Example 11 of this invention, HPLC purity 99.54% 1.3 Reagents and Instruments: Modified RPMI-1640 culture medium, penicillin-streptomycin solution, 0.25% trypsin-EDTA, fetal bovine serum, MTT, dimethyl sulfoxide, CO-150 carbon monoxide incubator, SW-CJ-2F medical clean bench, CKX-41-32 inverted microscope, CU600 electric thermostatic water bath, RT-2100C enzyme-linked immunosorbent assay (ELISA) analyzer. 2 Experimental Methods 2.1 Reagent Preparation 2.1.1 Preparation of MTT: Weigh 0.25g of MTT using a precision balance and place it in a 50mL volumetric flask. Add an appropriate amount of PBS, incubate in a 50-60℃ water bath, and shake well to dissolve completely. Add PBS to the mark to prepare a 5mg / kg solution. Filter the solution through a 0.22μm microporous membrane for sterilization, aliquot, and store in a refrigerator at 4℃ protected from light.

[0094] 2.1.2 Preparation of cell cryopreservation solution: Mix 20% serum, 10% DMSO and 70% 1640 medium evenly and store at -20℃.

[0095] 2.1.3 In Example 11 of the present invention, the compound was prepared by using DMSO to prepare a stock solution of the drug, and then diluted with culture medium to the concentration of the drug to be used. The final concentration of DMSO was controlled to be ≤0.1%.

[0096] 2.2 LO2 cell culture: Normal human LO2 cells were placed in a 25cm² culture medium. 2 Add approximately 4–5 mL of RPMI-1640 culture medium containing 10% FBS to cell culture flasks and incubate at 37°C in a 5% CO2 saturated humidity cell culture incubator. Change the culture medium every 2 days and observe cell growth daily. Once cells reach 80% confluence, passage or cryopreserve them. Use cells from passages 5–7 for formal experiments.

[0097] 2.3 Grouping and Drug Administration Experiment: The experiment was divided into a normal cell control group and drug administration groups with different concentrations of the present invention, based on the preliminary experimental results. The concentrations were 16.2, 32.4, 64.8, 130, 260, 518, 1036, 2073, 3240, and 4050 μmol / L, respectively.

[0098] 2.4 Hepatocyte MTT assay: Logarithmic growth phase LO2 cells were prepared into a cell suspension of 5.0 × 10³ cells / mL and seeded into 96-well plates. A normal control group and groups treated with different concentrations of the compound of the present invention (16.2, 32.4, 64.8, 130, 260, 518, 1036, 2073, 3240, and 4050 μmol / L) were included. After 24 h of culture, the culture medium was discarded, and the cells were washed 2–3 times with PBS. Different concentrations of the compound of the present invention were added, with 8 replicates per concentration. The plates were incubated at 37°C in a 5% CO₂ incubator. After 24, 48, and 72 h of culture, MTT solution was added at each time point, and the plates were incubated in the dark for 4 h. After 4 h, the supernatant was discarded, and 150 LDMSO was added to each well to dissolve the thiazolyl blue crystals. The plates were gently shaken to ensure uniform dissolution. The absorbance of each well was measured at 490 nm using a microplate reader. The absorbance value per well reflects the number of cells, and the two are directly proportional. The experiment was repeated three times. Cell viability was calculated as follows: Cell viability (%) = Absorbance of each group (OD490) × 100 / Absorbance of the control group (OD490) 2.5 Statistical Analysis All data are expressed as mean ± standard deviation and were processed using SPSS 17.0 statistical software. t-tests were performed for statistical analysis; paired t-tests were used for self-comparisons, and unpaired t-tests were used for inter-group comparisons. The significance level was P < 0.05.

[0099] 3. Experimental Results

[0100] Conclusion: After 72 h of administration, the OD values ​​of cells in each group showed an increasing trend at different concentrations of the present invention, but the growth rate decreased, indicating that the cells were still in the growth stage and the growth rate was somewhat inhibited. Different concentrations of the present invention had no inhibitory effect on cells; compared with the normal group, the cell survival rate was greater than 90%, and within the range of 16.2–4050 μmol / L, it showed almost no inhibition of cell growth and no cytotoxicity.

[0101] Test Example 5: Pharmacokinetic Study in Rats.

[0102] 1. Test sample and reference sample

[0103] 2. In vivo high performance liquid chromatography analysis method

[0104] 3 Experimental Design 3.1 Random Grouping This study selected 15 healthy SD rats with suitable SPF grades and randomly divided them into 5 groups, with 3 rats in each group. The test group was injected intramuscularly with YPK-033, while the control group was administered nintedanib tablet suspension by gavage (the rats were fasted for approximately 12 hours before administration, and given food and free access to water 1 hour after administration). Administration routes: intramuscular injection (hind leg muscle) and oral administration (gavage); frequency: single dose. 3.2 Sample Collection Collection method: Blood is collected from the submandibular vein, approximately 0.15 mL each time.

[0105] Data collection time points: Animals in the test group were collected at the following times: 0h before administration, 5min, 15min, 30min, 1h, 2h, 4h, 8h, 12h, 24h (1 day), 48h (2 days), 72h (3 days), 120h (5 days), 168h (7 days), 336h (14 days), 504h (21 days), and 672h (28 days).

[0106] Animals in the control group were monitored at 0 h before administration, 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h (1 day), and 48 h (2 days) after administration.

[0107] 3.3 Sample Separation and Preservation Sampling requirements and processing: Blood should be collected using EDTA-K2 anticoagulant tubes. Centrifuge to separate the plasma as soon as possible after collection. Store samples at approximately -20°C for further analysis; discard after analysis.

[0108] Conclusion: The compound of this invention exhibits stable blood drug release, remaining within the effective therapeutic concentration range for 672 hours. In contrast, similar acetates, decanoates, and hexadecano palmitates show a burst release effect, indicating that similar acetates, decanoates, and hexadecano palmitates possess potential therapeutic toxicity compared to long-term treatment tablets. Therefore, the compound of this invention has excellent application prospects.

[0109] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The compound represented by formula (Ⅰ) or a pharmaceutically acceptable salt thereof: 。 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The X-ray powder diffraction pattern of the compound represented by Formula I, expressed as a diffraction angle of 2θ±0.2°, is at 6.52±0.2°, 7.88±0.2°, 8.70±0.2°, 10.12±0.2°, 11.68±0.2°, 12.60±0.2°, 13.14±0.2°, 15.60±0.2°, 15.88±0.2°, 16.34±0.2°, 17.46±0.2°, 17.80±0.2°, 18.80±0.2°, 19.42±0.2°, 19.74±0.2°, and 20.34±0.2°. Characteristic peaks are present at 2°, 21.72±0.2°, 22.20±0.2°, 23.18±0.2°, 23.94±0.2°, 24.82±0.2°, 25.72±0.2°, 26.52±0.2°, 26.80±0.2°, 26.90±0.2°, 26.96±0.2°, 27.70±0.2°, 28.06±0.2°, 28.60±0.2°, 29.04±0.2°, 29.54±0.2°, 29.84±0.2°, 29.96±0.2°, and 30.78±0.2°.

3. The crystal form of the compound of formula I as described in claim 2, and its X-ray powder diffraction pattern are shown in Figure 2.

4. A method for preparing the nintedanib derivative of formula I as described in claim 1, or its solvate, or its pharmaceutically acceptable salt, characterized in that: Includes the following steps:

1. Nydanib was prepared as crude product (I) by reacting chloromethyl lauryl ester with a catalyst and solvent; The reaction formula is as follows: 。 5. A method for preparing the nintedanib derivative of formula I as described in claim 4, or its solvate, or its pharmaceutically acceptable salt, characterized in that: The reaction solvent in step 1 is selected from one or more of N,N-dimethylformamide, N,N-dimethylacetamide, DMSO, acetonitrile, and tetrahydrofuran, preferably acetonitrile; the catalyst is selected from cesium carbonate, pyridine, and DMAP, preferably cesium carbonate.

6. A pharmaceutical composition comprising a nintedanib derivative or a solvate thereof as shown in Formula I according to any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition of claim 6, formulated for intramuscular, intradermal, or subcutaneous injection.

8. Use of the compound of formula (I) of claim 1 or a salt thereof or the pharmaceutical composition of claim 6 in the preparation of a medicament for treating systemic sclerosis-related interstitial lung disease, idiopathic pulmonary fibrosis, and non-small cell lung cancer.

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