Pemetrexed disodium liquid composition, its preparation method and application

Abstract

The present invention discloses a pemetrexed disodium liquid composition, its preparation method and application. The pemetrexed disodium liquid composition provided by the present invention comprises a pharmaceutical active ingredient and a stabilizer, the pharmaceutical active ingredient being one or more of pemetrexed disodium, pharma- ceutically acceptable complexes, salts, solvates and hydrates of pemetrexed disodium, and the stabilizer being one or more of an organic solvent, a pH adjuster and an antioxidant. The pemetrexed disodium liquid composition of the present invention has the advantages of a simple preparation process, good physical and chemical stability, clinical ease of use without the need for reconstitution, low risk of microbial contamination during the preparation process, and suitability for industrial production.

Description

[Technical field]

[0001] This application claims priority to a prior application, bearing patent application number 202210645147.0 and entitled "Liquid composition of pemetrexed disodium, its preparation method and application," filed with the State Intellectual Property Office of the People's Republic of China on June 9, 2022. The above-mentioned prior application is incorporated herein by reference in its entirety. The present invention relates to a pemetrexed disodium liquid composition, its preparation method and application. [Background technology]

[0002] Pemetrexed is a multi-target antimetabolite antitumor drug, a folate antagonist that can inhibit folate-dependent enzymes such as thymidylate synthase, dihydrofolate reductase, and glycine ribonucleoside formyltransferase, which are involved in the biosynthesis of thymidine and purine nucleosides, thereby achieving its antitumor effect. Pemetrexed disodium was first developed by Eli Lilly and Company, and received marketing approval from the FDA in 2004. It obtained the Chinese import drug license in August 2005 and began clinical use in China. The drug is used in combination with cisplatin as a first-line drug to treat unresectable malignant pleural mesothelioma, and as a second-line drug to treat non-squamous non-small cell lung cancer as a single agent.

[0003] Currently available in several countries and regions around the world, the drug is a lyophilized powder injection that must be reconstituted with 0.9% sodium chloride solution to a concentration of 25 mg / mL according to pharmaceutical specifications before use, and then diluted a second time before administration. The reconstitution process may lead to dosage errors for patients, raising safety concerns and the risk of microbial contamination.

[0004] Therefore, finding a pemetrexed formulation that is easy to use, has good physical and chemical stability, has a simple manufacturing process, and is suitable for industrial production is currently a technical problem that needs to be solved urgently. Summary of the Invention

[0005] The pemetrexed disodium liquid composition provided by the present invention comprises a pharmaceutical active ingredient and a stabilizer, the pharmaceutical active ingredient being one or more selected from pemetrexed disodium, and pharma- ceutically acceptable complexes, salts, solvates and hydrates of pemetrexed disodium, and the stabilizer being one or more selected from organic solvents, pH adjusters and antioxidants.

[0006] According to an embodiment of the present invention, the content of the pharmaceutical active ingredient is 1 mg / mL to 50 mg / mL, preferably 15 mg / mL to 30 mg / mL, for example, 20 mg / mL, 25.00 mg / mL, 27.57 mg / mL, 30 mg / mL, 40 mg / mL, and the content refers to the ratio of the mass of the pharmaceutical active ingredient to the total volume of the pemetrexed disodium liquid composition.

[0007] According to an embodiment of the present invention, the organic solvent is one or more selected from ethanol, propylene glycol, and polyethylene glycol. For example, the polyethylene glycol is selected from polyethylene glycol 400 (PEG400) and / or polyethylene glycol 300 (PEG300).

[0008] According to some embodiments of the present invention, the content of the organic solvent is 70 mg / mL to 300 mg / mL, for example 90 mg / mL to 260 mg / mL, illustratively 90 mg / mL, 180 mg / mL, 260 mg / mL or 280 mg / mL, and the content refers to the ratio of the mass of the organic solvent to the total volume of the pemetrexed disodium pharmaceutical composition.

[0009] According to an embodiment of the present invention, the pH adjuster may be an acidic pH adjuster and / or a basic pH adjuster. For example, the acidic pH adjuster is preferably hydrochloric acid and / or citric acid, more preferably citric acid. For example, the basic pH adjuster may be one or more of sodium hydroxide, lysine, arginine, meglumine, and tromethamine, preferably tromethamine. For example, the lysine may be L-lysine and / or D-lysine. For example, the arginine may be L-arginine and / or D-arginine.

[0010] According to an embodiment of the present invention, the antioxidant is one or more selected from anhydrous sodium sulfite, cysteine ​​hydrochloride, acetylcysteine, and methionine, more preferably cysteine ​​hydrochloride. The content of the antioxidant is preferably 0-15 mg / mL, more preferably 0.1 mg / mL-10 mg / mL, for example, 0.3 mg / mL, 1 mg / mL, 1.63 mg / mL, or 3 mg / mL, and the content refers to the ratio of the mass of the antioxidant to the total volume of the pemetrexed disodium pharmaceutical composition.

[0011] In some embodiments, the content of the stabilizer (e.g., a basic pH adjuster) in the pemetrexed disodium liquid composition is preferably 0.5 mg / mL to 50 mg / mL, more preferably 1 mg / mL to 10 mg / mL, for example, 2.4 mg / mL, and the content refers to the ratio of the mass of the stabilizer (e.g., a basic pH adjuster) to the total volume of the pemetrexed disodium liquid composition.

[0012] According to some embodiments of the present invention, the stabilizer comprises at least one or more of citric acid, tromethamine, cysteine ​​hydrochloride, meglumine, and arginine, and in some further embodiments, the stabilizer may further comprise propylene glycol.

[0013] According to an embodiment of the present invention, the pH of the pemetrexed disodium liquid composition is preferably 7.5 to 9.5, and more preferably 8.0 to 9.0. According to an embodiment of the present invention, the pemetrexed disodium liquid composition may further contain an osmotic pressure adjusting agent. The osmotic pressure adjusting agent is preferably one or more selected from sodium chloride, mannitol, glycerin, and propylene glycol. The content of the osmotic pressure adjusting agent is preferably 1 mg / mL to 300 mg / mL, for example, 1 mg / mL to 100 mg / mL, more preferably 2 mg / mL to 30 mg / mL, for example, 5.8 mg / mL, 11 mg / mL, 15 mg / mL, or 23 mg / mL, and the content refers to the ratio of the mass of the osmotic pressure adjusting agent to the total volume of the pemetrexed disodium liquid composition. When the pemetrexed disodium liquid composition contains propylene glycol, the propylene glycol can not only adjust the osmotic pressure, but also improve the stability of the pemetrexed disodium liquid composition.

[0014] According to an embodiment of the present invention, the pemetrexed disodium liquid composition further comprises water, such as water for injection, to a volume of 1 mL. Illustratively, water is added to a volume of 1 mL.

[0015] According to an embodiment of the present invention, the pemetrexed disodium liquid composition is selected from any one of the following compositions:

[0016] Composition 1 comprises a pharmaceutical active ingredient, a stabilizer, and water, wherein the pharmaceutical active ingredient is pemetrexed disodium, a pharma- ceutically acceptable complex, salt, or hydrate thereof, and the stabilizer is selected from tromethamine and citric acid; Preferably, in the composition, the content of the active pharmaceutical ingredient is 25 mg / mL, and the content of the tromethamine is 2.4 mg / mL; Composition 2 comprises a pharmaceutical active ingredient, a stabilizer, and water, wherein the pharmaceutical active ingredient is pemetrexed disodium, a pharma- ceutically acceptable complex, salt, or hydrate thereof, and the stabilizer comprises tromethamine and citric acid, and further comprises anhydrous sodium sulfite, acetylcysteine, sodium thiosulfate, and / or cysteine ​​hydrochloride; Preferably, in the above composition II, the content of the medicament active ingredient is 20 mg / mL, 25 mg / mL, 27.57 mg / mL, 30 mg / mL or 40 mg / mL, the content of the above tromethamine is 2.4 mg / mL, the content of the above anhydrous sodium sulfite is 1 mg / mL, the content of the above acetylcysteine ​​is 1.63 mg / mL, the content of the above cysteine ​​hydrochloride is 0.3 mg / mL, and the content of the above sodium thiosulfate is 1 mg / mL; Composition 3: A composition comprising a pharmaceutical active ingredient, a stabilizer, and water, wherein the pharmaceutical active ingredient is pemetrexed disodium, a pharma- ceutically acceptable complex, salt, or hydrate thereof, and the stabilizer comprises citric acid and cysteine ​​hydrochloride, and further comprises meglumine and / or arginine; Preferably, in the above composition III, the content of the active pharmaceutical ingredient is 20 mg / mL, 25 mg / mL, 27.57 mg / mL, 30 mg / mL or 40 mg / mL, the content of the above cysteine ​​hydrochloride is 0.3 mg / mL, the content of the above meglumine is 2.4 mg / mL, and the content of the above arginine is 2.4 mg / mL; Composition 4: A pharmaceutical composition comprising a pharmaceutical active ingredient, a stabilizer, and water, wherein the pharmaceutical active ingredient is pemetrexed disodium, a pharma- ceutically acceptable complex, salt, or hydrate thereof, the stabilizer comprises tromethamine and citric acid, and further comprises propylene glycol and / or polyethylene glycol (e.g., polyethylene glycol 300 or polyethylene glycol 400); Preferably, in the above composition IV, the content of the medicament active ingredient is 20 mg / mL, 25 mg / mL, 27.57 mg / mL, 30 mg / mL or 40 mg / mL, the content of the above tromethamine is 2.4 mg / mL, the content of the above propylene glycol is 70 to 300 mg / mL (e.g., 90, 180, 260 mg / mL), and the content of the above polyethylene glycol is 70 to 300 mg / mL (e.g., 90, 180, 280 mg / mL); Composition 5: A composition comprising a pharmaceutical active ingredient, a stabilizer, an osmolality modifier, and water, wherein the pharmaceutical active ingredient is pemetrexed disodium, a pharma- ceutically acceptable complex, salt, or hydrate thereof, the stabilizer comprises tromethamine and citric acid, and the osmolality modifier is selected from sodium chloride, mannitol, glycerin, and / or propylene glycol; Preferably, in the fifth composition, the content of the active pharmaceutical ingredient is 20 mg / mL, 25 mg / mL, 27.57 mg / mL, 30 mg / mL or 40 mg / mL, the content of the tromethamine is 2.4 mg / mL, the content of the sodium chloride is 5.8 mg / mL, the content of the mannitol is 23 mg / mL, the content of the glycerin is 15 mg / mL, and the content of the propylene glycol is 11 mg / mL; Any one of the above compositions has a pH of 8.0 to 9.0.

[0017] The present invention further provides a method for preparing the pemetrexed disodium liquid composition, which may include a nitrogen filling process, which can be selected from liquid blending type nitrogen filling and / or container filling type nitrogen filling, and is preferably liquid blending type nitrogen filling and container filling type nitrogen filling. In some embodiments, nitrogen is filled so that the dissolved oxygen content in the pemetrexed disodium liquid composition is less than 5 mg / L, preferably less than 3 mg / L, and the dissolved oxygen content refers to the mass ratio of oxygen gas to the volume of the pemetrexed disodium liquid composition. In some embodiments, the head space residual oxygen content is less than 6%, preferably less than 4%, and the head space residual oxygen content refers to the volume fraction of oxygen gas in the mixed gas at the top of a sealed packaging container with which the pemetrexed disodium liquid composition directly contacts.

[0018] According to an embodiment of the invention, the manufacturing method includes non-terminal sterilization.

[0019] The present invention further provides the application of said pemetrexed disodium liquid composition in the preparation of antineoplastic drugs.Preferably, said antineoplastic drugs can be liquid drug formulations, such as oral liquids or injection liquids.

[0020] The present invention further provides an antitumor drug, which comprises the above-mentioned pemetrexed disodium liquid composition or is prepared from the above-mentioned pemetrexed disodium liquid composition. For example, the antitumor drug may be an oral liquid or an injectable liquid.

[0021] The present invention further provides a method for treating tumors, comprising providing a therapeutically effective amount of the pemetrexed disodium liquid composition or the antitumor drug to a patient in need thereof. Each of the above preferred conditions can be arbitrarily combined to obtain each of the relatively preferred embodiments of the present invention without departing from the common knowledge in the art.

[0022] All of the reagents and raw materials used in the present invention are commercially available.

[0023] Beneficial Effects of the Invention The pemetrexed disodium liquid composition of the present invention has the advantages of being simple in manufacturing process, having good physical and chemical stability, being clinically convenient without the need for reconstitution, having a low risk of microbial contamination during the preparation process, and being suitable for industrial production.

[0024] In the prior art, lyophilized powder of pemetrexed for injection needs to be reconstituted and then diluted twice before use, which effectively overcomes the drawbacks such as complicated operations and the reconstitution process that may cause differences in dosage to patients and the risk of microbial contamination. [Brief description of the drawings]

[0025] [Figure 1] 1 is a mean drug-time graph (N=6) following intravenous injection of Compound 16 and control Alimta® in Beagle dogs; [ka] shows the mean drug-time curve for formulation 16, [ka] shows the mean drug-time curves following the control drug Alimta®. [Diagram 2] This is the change trend of impurity PMQS-IM-B in the accelerated experiment of blend 16. [Diagram 3] This is the change trend of impurity PMQS-IM-C in the accelerated experiment of blend 16. [Figure 4] This is the change trend of impurity PMQS-IM-F in the accelerated experiment of blend 16. [Diagram 5] This is the change trend of impurity PMQS-IM-B in the long-term experiment of formulation 16. [Figure 6] This is the change trend of impurity PMQS-IM-C in the long-term experiment of formulation 16. [Figure 7]This is the change trend of impurity PMQS-IM-F in the long-term experiment of formulation 16. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] The present invention will be further described below with reference to examples, but the present invention is not limited to the scope of the described examples. In the following examples, experimental methods without specific conditions are selected according to conventional methods and conditions or product instructions.

[0027] The amount of pemetrexed disodium in the formulations in the Examples and Comparative Examples is calculated based on anhydrous pemetrexed disodium.

[0028] Comparative Example 1 [Table 1] The drug solution was prepared according to the amount of Formulation 1, and the drug solution was pre-filtered using a 0.22 μm filter core, and the resulting solution was filled into washed and sterilized vials and sealed.

[0029] Example 1 [Table 2] The drug solution was prepared according to the amount of formulation 2, and the drug solution was pre-filtered using a 0.22 μm filter core. The resulting solution was filled into a washed and sterilized vial, which was then purged with nitrogen gas to a headspace residual oxygen of <4%, and sealed.

[0030] Example 2 The samples obtained from formulations 1 and 2 were placed under high temperature / light illumination conditions (high temperature 40°C ± 2°C / 75% ± 5% RH, light illumination total illuminance 1.2 × 10 5 lux·hr, near UV energy 24 w·hr / m 2 ) The stability of the system was examined, and the appearance, pH, and related substances of the sample solutions were measured, and the data are summarized as follows. [Table 3] As is clear from the results of the solution appearance, in formulation 1, the pemetrexed solution was directly filled into the container, and the appearance of the solution changed under high temperature and light illumination conditions, while in formulation 2, the stabilizers tromethamine and citric acid were added to the formulation, and the appearance of the solution changed only under light illumination.

[0031] As is clear from the pH results, the pH of Blend 1 decreased under high temperature and light illumination conditions, with the maximum decrease being 1.7, while the pH of Blend 2 was stable under high temperature and light illumination conditions.

[0032] As is clear from the results of related substances, the total impurities in Blend 1 increased obviously under high temperature and light illumination conditions, and the total impurities in Blend 2 increased, but the increase tendency was smaller than that of Blend 1.

[0033] As mentioned above, compared with Formulation 1, after adding stabilizers and filling with nitrogen, the physical and chemical stability of the solution of Formulation 2 is obviously increased.

[0034] Example 3 [Table 4] The drug solutions were prepared according to the amounts of formulations 3 to 7, and the drug solutions were pre-filtered using a 0.22 μm filter core. The resulting solutions were filled into cleaned and sterilized vials, which were then purged with nitrogen gas to a headspace residual oxygen of <4%, and sealed.

[0035] Example 4 The samples obtained from formulations 3 to 5 were placed under high temperature / light illumination conditions (high temperature 40°C ± 2°C / 75% ± 5% RH, light illumination total illuminance 1.2 × 10 5 lux·hr, near UV energy 24 w·hr / m 2 ) The stability of the system was investigated, and the appearance, pH, and related substances of the sample solutions were measured. The data was compared with Formulation 2 and summarized as follows: [Table 5] As is clear from the results of the solution appearance, in formulation 2, no antioxidant was added, and the color of the system changed slightly after being left under light conditions for a while, while in formulations 3 to 5, different types of antioxidants were added, and after being left under high temperature and light for 5 and 10 days, the appearance did not change in any case.

[0036] As can be seen from the pH results, the pH values ​​of the systems are all relatively stable after different kinds of antioxidants are added.

[0037] As can be seen from the results of related substances, the total impurities of formula 2 (without antioxidant) remained basically unchanged under high temperature conditions; among formulas 3 to 5, the impurities of formula 3 (antioxidant is anhydrous sodium sulfite) and formula 4 (antioxidant is acetylcysteine) increased relatively quickly, but formula 5 (antioxidant is cysteine ​​hydrochloride) showed better stability, with single impurities below 0.06% and total impurities below 0.32%.

[0038] As mentioned above, in the formulation of antioxidant, cysteine ​​hydrochloride is added as an antioxidant to the pemetrexed solution, and the stability of the solution is relatively good. At the same time, the formulation without antioxidant can ensure excellent stability even under high temperature conditions, and the detection amount of single impurities is relatively low.

[0039] Example 5 [Table 6] The drug solution was prepared according to the amounts of formulations 8-9, and the drug solution was pre-filtered using a 0.22 μm filter core. The resulting solution was filled into a washed and sterilized vial, which was then purged with nitrogen gas to a headspace residual oxygen <4%, and sealed.

[0040] Example 6 The samples obtained from formulations 8 and 9 were placed under high temperature / light illumination conditions (high temperature 40°C ± 2°C / 75% ± 5% RH, light illumination total illuminance 1.2 × 105 lux·hr, near UV energy 24 w·hr / m 2 The stability of the system was examined, and the appearance, pH, and related substances of the sample solutions were measured. The data was compared with Formulation 5 (pH adjuster was tromethamine), and summarized as follows: [Table 7] As can be seen from the above results, when the pH adjusters meglumine, arginine, and tromethamine were added to the formulation, the appearance of the system did not change after the sample was placed under high temperature / light illumination conditions of 40°C for 5 and 10 days, and the single impurity was less than 0.08%, while the total impurities were only less than 0.33%, indicating that meglumine, arginine, and tromethamine have excellent stabilizing effects on pemetrexed solution and can function as stabilizers.

[0041] Example 7 [Table 8] The drug solution was prepared according to the amounts of formulations 10-12, and the drug solution was pre-filtered using a 0.22 μm filter core. The resulting solution was filled into a washed and sterilized vial, which was then purged with nitrogen gas to a headspace residual oxygen of <4%, and sealed.

[0042] The samples obtained from formulations 10 to 12 were placed under high temperature conditions (high temperature 40°C±2°C / 75%±5%RH) to examine the stability of the system, and the related substances in the sample solutions were measured. The data was compared with formulation 5 (pH adjuster is tromethamine), and summarized as follows. [Table 9] As can be seen from the above results, when the organic solvent propylene glycol is added to the formula at 180 mg / mL or more, the stability is increased similarly to that of formula 5 with the addition of antioxidants. It is illustrated that adding propylene glycol to the formula also has a stabilizing effect on the product.

[0043] Example 8 [Table 10] The drug solution was prepared according to the amounts of formulations 13-15, and the drug solution was pre-filtered using a 0.22 μm filter core. The obtained solution was filled into a washed and sterilized vial, and then purged with nitrogen gas to make the headspace residual oxygen <4%, and sealed.

[0044] Example 9 [Table 11] The drug solutions were prepared according to the amounts of formulations 16-19, and the drug solutions were pre-filtered using a 0.22 μm filter core. The resulting solutions were filled into cleaned and sterilized vials, and then purged with nitrogen gas to a headspace residual oxygen <4%, and sealed.

[0045] Example 10 The samples obtained from formulations 16 to 19 were placed under high temperature conditions (high temperature 40°C ± 2°C / 75% ± 5% RH) to examine the stability of the system, and the related substance data was summarized as follows. [Table 12] From the above table, it can be seen that the effect of different types of osmotic modifiers on the stability of the products is not clear.

[0046] In addition, sample solutions were prepared at the dilution ratios used in clinical use and their osmotic pressures were measured. The results are shown in the table below. [Table 13] The results show that all of the samples prepared in this example can meet the isotonicity requirements in the predicted clinical use.

[0047] Example 11 A drug solution was prepared according to the amounts of the formula 16, and the drug solution was filled into a colorless neutral borosilicate glass injection bottle, and the solution was filled with nitrogen gas to reduce the amount of dissolved oxygen. The oxygen content in the solution was measured, and the results are shown in the table below. [Table 14] The sample obtained after sealing the ampoule was placed under high temperature conditions (high temperature 40℃±2℃ / 75%±5%RH) to examine the stability of the system, and the data on appearance and related substances are summarized as follows. [Table 15] As can be seen from the above results, when the amount of dissolved oxygen in the solution is reduced, the amplification of total impurities in the pemetrexed disodium solution is further reduced, suggesting that the amount of dissolved oxygen in the solution has a certain effect on the stability of pemetrexed disodium. Therefore, in order to reduce the effect of the amount of dissolved oxygen in water, in the liquid blending process, first, nitrogen filling treatment is carried out on the water for injection with high purity nitrogen gas to replace the dissolved oxygen in water, so that the effect of dissolved oxygen on the pemetrexed disodium solution can be reduced.

[0048] Example 12 [Table 16] A drug solution was prepared according to the blending amounts in the above table, and the drug solution was filled into a colorless neutral borosilicate glass injection bottle. It was then examined under a head space whether or not filling with nitrogen had any effect on the stability.

[0049] The manufactured samples were placed under high temperature conditions (high temperature 40℃±2℃ / 75%±5%RH) to check the stability of the system, and the appearance of the sample solution and related substances were inspected. The results are shown in the table below. [Table 17] As can be seen from the results of the solution appearance, when the sample without nitrogen filling was placed under high temperature conditions for 10 days, the appearance of the solution began to change, and it transformed from a colorless and transparent liquid to a slightly yellowish and transparent liquid. However, after the headspace nitrogen filling process was applied, the solution systems with different residual oxygen contents were placed under high temperature conditions for different periods, and their appearances did not change obviously, and they were colorless and transparent solutions.

[0050] As is clear from the results of the related substances, the sample filled with headspace nitrogen had 0.31% impurities after 10 days at 40℃, which is 0.26% less than the sample not filled with headspace nitrogen. Therefore, the product must be filled with headspace nitrogen.

[0051] Formulation 5 was tested for headspace nitrogen filling to determine headspace residual oxygen limits. [Table 18] The drug solution was prepared according to the formulation in the above table, and the drug solution was pre-filtered using a 0.22 μm filter core. The resulting solution was filled into a washed and sterilized vial, and the effect of different nitrogen filling processes on stability was examined.

[0052] The samples with different residual oxygen contents were placed under high temperature conditions (high temperature 40℃±2℃ / 75%±5%RH) to check the stability of the system, and the appearance of the sample solution and related substances were inspected. The results are shown in the table below. [Table 19] The samples with headspace residual oxygen of 3.9% and 1.8% showed relatively small increases in total impurities, both close to the results for day 0, and no significant difference between the two groups.

[0053] As mentioned above, the nitrogen filling process has a relatively large effect on the stability of the system, and when the headspace residual oxygen is controlled to 4% or less, the stability of the system is relatively good.

[0054] Example 13 Using formula 16 and corresponding processes, i.e., before liquid blending, nitrogen was filled into the water for injection to replace oxygen gas, and then vacuum was drawn by freeze dryer and nitrogen was filled to control the headspace residual oxygen within 4%. Three batches of products, coded 1, 2, and 3, were commercially produced. The obtained three samples were placed under accelerated conditions (25℃ / 60%RH) and long-term conditions (5±3℃), respectively. The obtained related substance testing results are shown in the following Tables A-B and Figures 2-7. [Table 20] As can be seen from Table A and Figures 2-4, the 6-month stability data under accelerated conditions is well within the limits and meets the criteria. [Table 21] As can be seen from Table B and Figures 5 to 7, under long-term conditions (5±3°C), data from 6 months indicates that related substances are well within limits and stability is good.

[0055] From the above data, it can be seen that the pemetrexed disodium liquid composition of the present invention has good stability, the manufacturing process is stable, and it is suitable for commercial production.

[0056] Considering the general principle that the injection formulation should be as simple as possible, formulation 16 was selected for consideration in subsequent non-clinical studies.

[0057] Example 14 In this study, a total of 12 Beagle dogs, half male and half female, were used, and compound 16 and the control drug Alimta (registered trademark) were intravenously administered according to a two-cycle crossover experimental design. The doses of both were 25 mg / kg, and sampling times were before administration, immediately after the end of administration (0-1 min), 5 min, 20 min, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 10 h, 12 h, 16 h, 24 h, and 36 h. The average pharmacokinetic parameters of pemetrexed in the plasma of Beagle dogs in each group are shown in Table 1, and the average drug concentration-time graph is shown in Figure 1. [Table 22] As can be seen from the above results, Formulation 16 can achieve exposure levels close to those of the control drug, and the important pharmacokinetic parameters are essentially consistent.

[0058] Example 15 Equilibrium dialysis was used to investigate the binding rate of different concentrations of Formulation 16 (based on the pharmacokinetic results of Formulation 16 after intravenous injection in Beagle dogs, combined with the preclinical data of the reference formulation, the low, medium and high concentrations of the experimental groups were set at 10, 100 and 1000 μg / mL, respectively) to human plasma protein, and compared with the binding rate of the control drug Alimta (registered trademark). At concentrations of 10, 100 and 1000 μg / mL, Formulation 16 has a human plasma protein binding rate of 86.50%, 90.19% and 74.72%, respectively, and the control drug (Alimta (registered trademark)) has a human plasma protein binding rate of 86.14%, 89.94% and 80.96%, respectively, at the corresponding concentrations. The detailed data of the plasma protein binding rate are shown in Table 2 below. [Table 23] As a result, under the conditions of this study, the binding rates of pemetrexed disodium to human plasma proteins for the 10 μg / mL, 100 μg / mL, and 1000 μg / mL formulations 16 and the control drug Alimta (registered trademark) were all at moderate levels (50% to 90%).

[0059] Example 16 Pemetrexed disodium is a multi-target antimetabolic antineoplastic drug, which was clinically administered intravenously after dilution with 0.9% sodium chloride injection, and an in vitro hemolysis test was performed on formulation 16.

[0060] The study included a negative control group (0.9% sodium chloride injection, tube 1), a positive control group (sterile water for injection, tube 2), a control drug Alimta® group (pemetrexed disodium for injection, 10 mg / mL in dosage, 0.1, 0.2, 0.3, 0.4, and 0.5 mL / tube, tubes 3 to 7), and 16 combination groups (10 mg / mL in dosage, 0.1, 0.2, 0.3, 0.4, and 0.5 mL / tube, tubes 8 to 12). Each group consisted of three parallel tubes. 2% red blood cell suspension, 0.9% sodium chloride injection, sterile water for injection, control drug Alimta (registered trademark), and 16 combination groups were mixed uniformly in a set ratio, and then placed in an incubator at a temperature range of 37±0.5°C, and observed once each at 0, 15, 30, 45, 60, 120, and 180 minutes (±10%). The specific results are shown in Table 3. [Table 24] As a result, after leaving it at 37±0.5℃ for 180 minutes, the tube of the negative control group had all the red blood cells settled, the upper layer was colorless and transparent, and after shaking appropriately, the settled red blood cells were redispersed, and no hemolysis or aggregation was observed, while the tube of the positive control group had a clear red solution, no red blood cells remained at the bottom of the tube, indicating total hemolysis.The above results suggest that this test system is normal and reliable.

[0061] In the tubes of the Alimta (registered trademark) group, all red blood cells had settled, the upper layer was colorless and transparent, and no hemolysis was observed. However, among the settled red blood cells, agglutinated red blood cells were also found, which did not disperse even when shaken, and the agglutinated red blood cells did not disperse even upon microscopic examination, and it was determined that the tubes were not hemolyzed but were agglutinated.

[0062] In the tubes containing formulation 16, all red blood cells had settled, the upper liquid was colorless and transparent, and after appropriate shaking the settled red blood cells were redispersed. Microscopic examination revealed no abnormalities in the red blood cells, and no hemolysis or aggregation was observed.

[0063] As mentioned above, the 16th formulation at a concentration of 10 mg / mL had no hemolytic or agglutinating effect on rabbit red blood cells, and the results of the in vitro hemolysis test were negative.

[0064] Example 17 For clinical use, this product needs to be diluted with 0.9% sodium chloride injection and then injected intravenously. An active systemic anaphylaxis test was conducted on guinea pigs with Formulation 16 to observe whether anaphylaxis occurred after administering the test product to guinea pigs, and this was used as a reference for evaluating the safety of clinical use.

[0065] Six groups were set up in this study, namely, a negative control group, a positive control group, an Alimta® low-dose group, a positive control group, and a low-dose group and a high-dose group of Formula 16, each of which consisted of six guinea pigs, half male and half female. In the sensitization stage, the negative control group was intravenously injected with 0.9% sodium chloride injection at a volume of 4 mL / kg, the positive control group was intraperitoneally injected with 8 mg / mL chicken egg albumin solution at a volume of 0.5 mL / animal, and the Alimta® and Formula 16 low-dose and high-dose groups were intravenously injected with 10 mg / mL Alimta® or Formula 16 at a volume of 2 mL / kg and 4 mL / kg, respectively, with doses of 20 mg / kg and 40 mg / kg, which are approximately 1.5 and 3 times the maximum dose to be used in clinical trials. The day of the first administration was defined as the first day of the study. Sensitization was performed once every other day, and three consecutive sensitizations were performed (i.e., sensitization was performed on the first, third, and fifth days of the study). On the 14th and 21st days after the final sensitization (i.e., the 19th and 26th days of the study), the guinea pigs were challenged by intravenous injection of twice the sensitizing dose, and the systemic reactions and deaths of the guinea pigs after challenge were observed. [Table 25] [Table 26] The results showed that during the sensitization period, the guinea pigs in each group were generally in good condition, with normal spontaneous movement, clean skin and fur, no abnormal secretions, normal weight gain, and no other abnormal symptoms.

[0066] Within 30 minutes after the initial induction, none of the guinea pigs in the negative control group, the Alimta (registered trademark) low and high dose groups, and the Formulation 16 low and high dose groups showed anaphylaxis, whereas the guinea pigs in the positive control group showed anaphylactic symptoms and all died within 7 minutes after administration, indicating strongly positive anaphylaxis.

[0067] Within 30 minutes after the final stimulation, none of the guinea pigs in the negative control group, the Alimta® low and high dose groups, or the Formulation 16 low and high dose groups experienced anaphylaxis.

[0068] As described above, in the present study, British guinea pigs were intravenously injected with 10 mg / mL of formulation 16 at a volume of 2 and 4 mL / kg, respectively, i.e., doses of 20 and 40 mg / kg, and the results of the active systemic anaphylaxis test were negative.

[0069] Example 18 A single intravenous injection irritation test was conducted on formulation 16 in rabbits to observe the irritation reaction and reversibility of the test substance on the rabbit ear marginal vein blood vessels and surrounding tissues, and use the results as a reference for the safety evaluation of clinical use.

[0070] This study included two groups, an Alimta® group and a Combination 16 group, each of which contained eight rabbits, half male and half female. The rabbits in each group were intravenously injected with 10 mg / mL Alimta® or Combination 16 at a volume of 4 mL / kg through the right ear margin, resulting in a dose of 40 mg / kg, which is approximately three times the maximum dose to be used in clinical trials. At the same time, an autologous control was used, and an equal volume of 0.9% sodium chloride injection was intravenously injected through the left ear margin. A single dose was administered, and the rabbits were continuously observed for 21 days after administration. The day of administration was defined as the first day of the study.

[0071] During the test period, the general condition of the rabbits and the injection sites were observed every day. On the 3rd and 21st days after administration (i.e., the 4th and 22nd days of the test), 4 rabbits (half male and half female) from each group were euthanized and dissected to visually observe the local irritation reaction after injection, and the tissues taken were pathologically examined.

[0072] During the observation period after administration, none of the rabbits in each group showed any reactions such as redness, swelling, congestion or necrosis at either injection site. The rabbits were in good general condition, their spontaneous movements were normal, and no other abnormal symptoms were observed.

[0073] On days 3 and 21 after administration, no visible abnormal changes were observed at either injection site in any of the rabbits in each group.

[0074] Three days after administration, mild vascular inflammation associated with mechanical irritation of the injection puncture appeared at one of the injection sites on the control side (1 / 8).Otherwise, no abnormalities were observed at any of the other injection sites in each group on days 3 and 21 after administration.

[0075] As is evident from the above results, under the conditions of this study, when 10 mg / mL of Formulation 16 was intravenously injected into Japanese White rabbits via the right ear margin at a volume of 4 mL / kg, i.e., a dose of 40 mg / kg, no irritation was observed in the blood vessels and surrounding tissues at the injection site.

[0076] As described above, the pemetrexed disodium liquid composition provided by the present invention can achieve an exposure level close to that of commercially available control products, and is less irritating to the body, has fewer adverse reactions, is safer, and is easier to use, thereby improving patient compliance and clinical dosing convenience, and has promising prospects for commercialization.

[0077] Although several embodiments of the present invention have been described in detail, those skilled in the art may make various improvements and modifications to the specific embodiments described without substantially departing from the teachings and advantages of the subject matter of the present invention, and such improvements and modifications are within the spirit and scope of the present invention as encompassed by the appended claims.

Claims

1. A pemetrexed disodium liquid composition comprising an active pharmaceutical ingredient and a stabilizer, wherein the active pharmaceutical ingredient is one or more of pemetrexed disodium, and pharma- ceutically acceptable complexes, salts, solvates, and hydrates of pemetrexed disodium, and the stabilizer is one or more of an organic solvent, a pH adjuster, and an antioxidant. A pemetrexed disodium liquid composition comprising:

2. the content of the active pharmaceutical ingredient is 1 mg / mL to 50 mg / mL, and the content refers to the ratio of the mass of the active pharmaceutical ingredient to the total volume of the pemetrexed disodium liquid composition; The pemetrexed disodium liquid composition of claim 1.

3. the content of the active pharmaceutical ingredient is 15 mg / mL to 30 mg / mL, and the concentration refers to the ratio of the mass of the active pharmaceutical ingredient to the total volume of the pemetrexed disodium liquid composition; and / or the organic solvent is one or more of ethanol, propylene glycol, and polyethylene glycol; and / or The pH adjuster is an acidic pH adjuster and / or a basic pH adjuster, and / or The antioxidant is one or more of anhydrous sodium sulfite, cysteine ​​hydrochloride, acetylcysteine, and methionine. The pemetrexed disodium liquid composition according to claim 1 or 2.

4. The acidic pH adjuster is hydrochloric acid and / or citric acid; and / or the basic pH adjuster being one or more of sodium hydroxide, lysine, arginine, meglumine, and tromethamine; and / or The polyethylene glycol is polyethylene glycol 300 and / or polyethylene glycol 400. The pemetrexed disodium liquid composition of claim 3.

5. The lysine is L-lysine and / or D-lysine, and / or The arginine is L-arginine and / or D-arginine. The pemetrexed disodium liquid composition of claim 4.

6. In the pemetrexed disodium liquid composition, the content of the basic pH adjusting agent is 0.5 mg / mL to 50 mg / mL, and the content refers to the mass ratio of the basic pH adjusting agent to the total volume of the pemetrexed disodium liquid composition; and / or The pemetrexed disodium liquid composition has a pH of 7.5 to 9.5; and / or The content of the antioxidant is 0-15 mg / mL, and the content refers to the ratio of the mass of the antioxidant to the total volume of the pemetrexed disodium liquid composition; The pemetrexed disodium liquid composition of claim 3.

7. In the pemetrexed disodium liquid composition, the content of the basic pH adjusting agent is 1 mg / mL to 10 mg / mL, and the content refers to the mass ratio of the basic pH adjusting agent to the total volume of the pemetrexed disodium liquid composition; and / or The pH of the pemetrexed disodium liquid composition is 8.0 to 9.0; and / or the content of the antioxidant is 0.1 mg / mL to 10 mg / mL, and the content refers to the ratio of the mass of the antioxidant to the total volume of the pemetrexed disodium liquid composition; The pemetrexed disodium liquid composition of claim 6.

8. The pemetrexed disodium liquid composition further comprises a osmolality adjusting agent, Preferably, the osmolality adjusting agent is one or more of sodium chloride, mannitol, glycerin and propylene glycol; and / or the content of the osmotic agent is 1 mg / mL to 300 mg / mL, and the content refers to the ratio of the mass of the osmotic agent to the total volume of the pemetrexed disodium liquid composition; The pemetrexed disodium liquid composition according to any one of claims 1 to 7.

9. A method for preparing the pemetrexed disodium liquid composition according to any one of claims 1 to 8, comprising a nitrogen filling process, Preferably, the nitrogen filling process is liquid blending type nitrogen filling and / or container filling type nitrogen filling; Preferably, the nitrogen filling process is to fill with nitrogen so that the dissolved oxygen content in the pemetrexed disodium liquid composition is less than 5 mg / L and / or the head space residual oxygen content is less than 6%, and more preferably, the nitrogen filling process is to fill with nitrogen so that the dissolved oxygen content in the pemetrexed disodium liquid composition is less than 3 mg / L and / or the head space residual oxygen content is less than 4%. A manufacturing method comprising the steps of:

10. The application of the pemetrexed disodium liquid composition according to any one of claims 1 to 8 in the preparation of antineoplastic drugs, preferably, the antineoplastic drugs are oral liquids or injection liquids.

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