Dipeptide derivative composition, method for producing the same, and use

Abstract

The present invention relates to a dipeptide derivative composition, and its preparation and use, the composition comprising (a) a compound of formula I, (b) glycine and (c) an antioxidant, the composition has good compatibility with auxiliary agents, and a lyophilized preparation prepared based on the composition has low impurity content and good stability under high humidity, strong light and low temperature storage conditions.

Description

Detailed Description of the Invention

[0001] This application claims the priority of a prior application filed with the China National Intellectual Property Administration on May 6, 2022, with a patent application number of 202210532274.X and an invention title of "Dipeptide Derivative Composition, Its Manufacturing Method and Use". The entire content of the above application is incorporated herein by reference.

[0002] 〔Technical Field〕 The present invention belongs to the field of pharmaceutical preparations, and specifically relates to a dipeptide derivative composition, its manufacturing method and use.

[0003] 〔Background Art〕 Liver failure refers to severe liver damage caused by multiple factors, which causes severe impairment or decompensation in the functions of the liver itself, such as synthesis, detoxification, excretion, and in-vivo changes. Clinically, a group of syndromes represented by disorders of the thrombin mechanism, jaundice, hepatic encephalopathy, dehydration, etc. appear. The fatality rate of liver failure is extremely high.

[0004] Patent applications CN201110025509.8 and CN201110025516.8 disclose dipeptide derivatives that can be used in the treatment of liver failure, and their structures are as shown below. [[ID=二十一]] [[ID=二十二]]

[0005] [[ID=二十三]] [[ID=二十四]] [[ID=二十五]]

Chemical Formula

[0006] [[ID=三十一]] [[ID=三十二]]The chemical name of the dipeptide derivative is 3-(2-benzyloxycarbonylamino-3-methyl-butanamide)-5-fluoro-4-oxo-pentanoic acid (F573), which can significantly inhibit or reverse liver failure, has a significant therapeutic effect on liver failure, and has no obvious toxicity to cells. [[ID=三十三]] [[ID=三十四]]

[0007] [[ID=三十五]] The dipeptide derivative is easily destroyed by various enzymes in the oral and gastrointestinal environments, and its efficacy is easily lost due to the first-pass effect of the liver. Therefore, it is not suitable for designing an oral dosage form. Furthermore, research has shown that the dipeptide derivative has poor stability in aqueous solutions and cannot withstand moist heat sterilization. Therefore, there is a need to develop an injectable dosage form that is highly stable, has a low impurity content, and has few side effects.

[0008] [Summary of the Invention] In order to improve upon the problems present in the prior art, in a first embodiment, the present invention is (a) A compound of formula I, whose structure is as follows:

[0009] [ka]

[0010] , (b) Glycine, (c) Provides a pharmaceutical composition containing an antioxidant.

[0011] According to embodiments of the present invention, the antioxidant is one, two, or more selected from ascorbic acid, sodium edetate, sodium bisulfite, and sodium pyrosulfite.

[0012] According to embodiments of the present invention, the above composition optionally further comprises composition (d) a pH adjuster, in some embodiments the pH adjuster being an alkaline reagent, which is one, two or more selected from sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, or disodium hydrogen phosphate.

[0013] According to embodiments of the present invention, composition (a) accounts for about 0.5% to about 10.0% (w / w) of the total amount of the composition, for example, 0.5%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, and 10.0% (w / w). Preferably, composition (a) accounts for about 2.0% to about 5.0% (w / w) of the total amount of the composition. According to embodiments of the present invention, composition (b) accounts for about 0.5% to about 15.0% (w / w) of the total amount of the composition, for example, 0.5%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, and 15.0% (w / w). Preferably, composition (b) accounts for about 3.0% to about 10.0% (w / w) of the total amount of the composition. According to embodiments of the present invention, composition (c) accounts for about 0.01% to about 0.50% (w / w) of the total amount of the composition, for example, 0.01%, 0.05%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.40%, and 0.50% (w / w). Preferably, composition (c) accounts for about 0.05% to about 0.30% (w / w) of the total amount of the composition. According to embodiments of the present invention, the pH value of the composition is greater than 7.0, and in some embodiments, the pH value range of the composition is selected from 7.0 to 9.0, for example, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, and 9.0, and preferably 7.5 to 8.5.

[0014] According to embodiments of the present invention, the above composition is a lyophilized powder injection.

[0015] In a second embodiment, the present invention is (a1) Add the specified amount of antioxidant and glycine, and optionally add a pH adjuster, and mix uniformly. (a2) Add the compound of formula I in the appropriate amount to the solution obtained in step (a1) and dissolve it, The present invention provides a method for producing the above-mentioned pharmaceutical composition, which includes the above-mentioned [ingredient].

[0016] According to embodiments of the present invention, the method for producing the above-mentioned pharmaceutical composition is: (a3) Further comprising the steps of sterilizing and filtering the liquid obtained in step (a2) and freeze-drying it.

[0017] According to embodiments of the present invention, in step (a1) above, the antioxidant and glycine are added to the pH adjuster in the specified amounts, preferably in aqueous solution form with a concentration of 0.5 to 3 mol / L, for example, 0.5 mol / L, 1.0 mol / L, 1.5 mol / L, 2.0 mol / L, or 3.0 mol / L, and more preferably the pH adjuster is a 1 mol / L aqueous solution of sodium bicarbonate. In some embodiments, the pH value of the liquid obtained in step (a1) is greater than 7.0, preferably in the range of 7.0 to 9.0, and more preferably 7.5 to 8.5. According to embodiments of the present invention, in step (a2) above, the temperature of the solution obtained in step (a1) is controlled to 8 to 15°C, and the compound of formula I is added in a further amount. In some embodiments, the compound of formula I is first sieved through an 80 to 200 mesh sieve (for example, it may be selected from a 100 mesh sieve), and then added to the solution obtained in step (a1). According to embodiments of the present invention, nitrogen gas protection is employed in the processes of steps (a1) and (a2) described above.

[0018] According to embodiments of the present invention, in step (a3) ​​above, a Millipore filter membrane is used for sterilization filtration. Preferably, the Millipore filter membrane is a polyethersulfone Millipore filter membrane.

[0019] According to one preferred method of the present invention, the method for producing the above-mentioned pharmaceutical composition is: Weigh the sodium pyrosulfite and glycine in the compounding amounts, add them to the sodium bicarbonate solution, stir, and completely dissolve them. Adjust the pH value to 7.0 - 9.0, control the solution temperature to 8 - 15 °C, then add the F573 in the compounding amount, stir and dissolve for 30 - 50 minutes, perform nitrogen gas protection during the whole liquid compounding process, sterilize and filter, fill, and at the same time Blooming add a butyl rubber stopper to an appropriate height and include the step of freeze-drying. Preferably, it further includes in-box plugging, out-of-box, capping, lamp inspection, labeling, packaging, sample submission, and warehousing of the finished product after passing the inspection after freeze-drying.

[0020] [[ID=[6]] In a third aspect, the present invention provides the use of the above pharmaceutical composition in the manufacture of a drug for preventing or treating liver failure. <​​​​​​​​​​​​​​​​​The technical aspects of the present invention will be described in more detail below, in accordance with specific embodiments. The embodiments described below are merely illustrative and should not be interpreted as limiting the scope of the claims of the present invention. Any technology realized based on the above-described aspects of the present invention falls within the scope of the claims of the present invention.

[0025] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be manufactured by known methods.

[0026] reagent 3-(2-benzyloxycarbonylamino-3-methylbutanamide)-5-fluoro-4-oxopentanoic acid (abbreviated as F573 and supplied by Beijing Condini Pharmaceutical Co., Ltd.) has the following structure:

[0027] [ka]

[0028] equipment equipment LYO-25 medical vacuum freeze dryer, Shimadzu 20A high-performance liquid chromatograph, HP1100 high-performance liquid chromatograph, Mettler AE240 balance.

[0029] (Example 1: Preparation of F573 Injectable Solution) 1.1 Screen injectable solvents for the purpose of manufacturing the F573 injectable solution.

[0030] [Table 1]

[0031] The results indicate that F573 completely dissolved in propylene glycol / aqueous solution (80% / 20%, pH adjusted to 4.0-5.0).

[0032] 1.2 Stability Considerations for F573 Injectable Solution F573 was dissolved in propylene glycol / aqueous solution (80% / 20%, pH adjusted to 4.0-5.0), sterilized at 120°C for 15 minutes, and then tested (see Table 1-2 below for results).

[0033] [Table 2]

[0034] The results show that in samples sterilized at high temperatures, the related substances increased by approximately 50%, while the F573 content decreased by approximately 50%. Since F573 has poor stability in aqueous solutions and cannot withstand moist heat sterilization, it is unsuitable for development into small-volume or large-volume injectable formulations.

[0035] (Example 2: Powdered injection (preparation using F573 + non-aqueous solvent)) To manufacture the F573 powder injection in composite packaging, each set contains one vial of sterile powder with 30 mg of the active ingredient and 2 mL of non-aqueous solvent. It is to be prepared and used immediately, with an appropriate amount of analgesic (benzyl alcohol) added as needed. First, a 90% ethanol solution is used as the solvent to dissolve, crystallize, decarburize, and dry the F573 raw material. The results show that performing processes such as recrystallization and decarburization on the raw material results in a yield of approximately 50% and can cause problems such as changes in crystal form. Therefore, F573 is not suitable for development in the above dosage form.

[0036] (Example 3: Study on the formulation of freeze-dried compositions) 3.1 Experiment on the effects of auxiliary agents In lyophilized formulations, mannitol can be used as a carrier to form a uniform skeleton, but amino acids can be used not only as skeleton agents for lyophilized materials but also as general protein protectants. The formulation is selected to be one in which no other auxiliary agents are added to F573, and another in which arginine, glycine, and mannitol are added, respectively. The lyophilized process involves sieving F573 through a 100-mesh sieve to prepare it for use. An appropriate amount of sodium bicarbonate is weighed and prepared as a 1 mol / L solution for use. The required amount of arginine, glycine, or mannitol is weighed and added to the sodium bicarbonate solution, stirred to dissolve completely, the solution temperature is controlled to 8-15°C, then the required amount of F573 is added, stirred while adding, and stirred for 40 minutes to dissolve. Nitrogen gas protection was maintained throughout the entire liquid formulation process. A 0.22 μm polyethersulfone Millipore filter membrane (filter cartridge) is used as the terminal filter for sterilization filtration and packing. Blooming Butyl rubber stoppers were added to the appropriate height, and freeze-dried to obtain a lump.

[0037] The research results show that glycine is even more suitable as a skeletal agent, as shown in Table B-1 below.

[0038] [Table 3]

[0039] 3.2 Compatibility tests between F573 and auxiliary agents were conducted, and studies were performed by combining F573 with glycine and sodium pyrosulfite. As shown in Tables B-2 and B-3 below, it was found that F573 is suitable for combination with glycine and sodium pyrosulfite.

[0040] [Table 4]

[0041] [Table 5]

[0042] 3.3 Optimization Experiment of Formulation Sodium pyrosulfite and glycine were selected as adjuvants for the F573 freeze-dried formulation. The formulation was further optimized by considering these two factors, setting each factor to three levels. Nine formulations were arranged using the orthogonal array L9(34), and experiments were conducted. The results were analyzed using the physical appearance of the freeze-dried needle, redissolution time, and pH value as evaluation indicators. The distribution of experimental factor levels is shown in Table C-1, and the experimental results and statistical analysis are shown in Tables C-2 and C-3.

[0043] [Table 6]

[0044] [Table 7]

[0045] [Table 8]

[0046] (Example 4: Freeze-dried formulation F-1) 4.1 Manufacturing of the F-1 Formulation F-1 (dosage for 1000 bottles) F573 30 g Glycine 50 g Sodium pyrosulfite 1 g Sodium bicarbonate (appropriate amount) Add 1000 mL of sterile water for injection.

[0047] F573 was sieved through a 100-mesh sieve and prepared for use. An appropriate amount of sodium bicarbonate was weighed and prepared into a 1 mol / L solution and prepared for use. Separately, the required amounts of sodium pyrosulfite and glycine were weighed and added to the sodium bicarbonate solution, stirred until completely dissolved, the pH was adjusted to 7.5-8.5, the solution temperature was controlled to 8-15°C, then the required amount of F573 was added, stirred while adding, and stirred for 40 minutes until dissolved. Nitrogen gas protection was maintained throughout the entire liquid mixing process. After inspection, pH and content passed, and the solution was filtered. A polyethersulfone millipore filter membrane (filter cartridge) was used as the final filter for sterilization filtration, and after the visible foreign matter inspection of the drug solution passed, it was placed in a large drug bottle, the amount to be charged was adjusted, and it was filled at the same time. Blooming Butyl rubber stoppers were added to the appropriate height. Freeze-drying, internal pressure sealing, unpacking, capping, lamp inspection, labeling, packaging, sample submission, and storage of finished products after inspection approval were carried out. Finished products were stored at 4-8°C.

[0048] 4.2 Three batches of samples were taken according to the F-1 manufacturing process described above, and the long-term stability of each formulation under high humidity, strong light, and low-temperature storage conditions was examined. The test results are as follows. The results show that each test item for Injectable F573 conforms to the pharmacopoeia's requirements for the stability of the formulation.

[0049] [Table 9]

[0050] [Table 10]

[0051] [Table 11]

[0052] [Table 12]

[0053] [Table 13]

[0054] [Table 14]

[0055] Illustrative embodiments of the present invention have been described above. However, the claims of the present invention are not limited to the embodiments described above. Any modifications, equivalent substitutions, improvements, etc., made without departing from the spirit and principles of the present invention should be included within the scope of the claims of the present invention.

Claims

1. A pharmaceutical composition, (a) A compound of formula I, whose structure is as follows: 【Chemistry 1】 、 (b) Glycine, (c) A composition containing antioxidants, Pharmaceutical composition.

2. The antioxidant is characterized by being one, two, or more selected from ascorbic acid, sodium edetate, sodium bisulfite, and sodium pyrosulfite. The pharmaceutical composition according to claim 1.

3. The aforementioned pharmaceutical composition further comprises composition (d) a pH adjuster. The pharmaceutical composition according to claim 1.

4. The pH adjusting agent is an alkaline reagent, and is characterized by being one, two, or more selected from sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, or disodium hydrogen phosphate. The pharmaceutical composition according to claim 3.

5. The above composition (a) accounts for 0.5% to 10.0% (w / w) of the total amount of the pharmaceutical composition, The aforementioned composition (b) accounts for approximately 0.5% to approximately 15.0% (w / w) of the total amount of the pharmaceutical composition. The composition (c) is characterized in that it accounts for 0.01% to 0.50% (w / w) of the total amount of the pharmaceutical composition. The pharmaceutical composition according to claim 1.

6. The composition (a) comprises 2.0% to 5.0% (w / w) of the total amount of the pharmaceutical composition. The pharmaceutical composition according to claim 5.

7. The composition (b) comprises 3.0% to approximately 10.0% (w / w) of the total amount of the pharmaceutical composition. The pharmaceutical composition according to claim 5.

8. The aforementioned composition (c) accounts for 0.05% to 0.30% (w / w) of the total amount of the pharmaceutical composition. The pharmaceutical composition according to claim 5.

9. The aforementioned pharmaceutical composition is characterized by having a pH value greater than 7.

0. The pharmaceutical composition according to claim 1.

10. The aforementioned pharmaceutical composition is characterized by having a pH value of 7.0 to 9.

0. The pharmaceutical composition according to claim 9.

11. The aforementioned pharmaceutical composition is characterized by having a pH value of 7.5 to 8.

5. The pharmaceutical composition according to claim 9.

12. Furthermore, it contains an amount of sterile water for injection with a total volume of 1000 mL. It contains 30g of composition (a), 50g of composition (b), and 1g of composition (c), and The aforementioned pharmaceutical composition is characterized by having a pH value of 7.5 to 8.

5. The pharmaceutical composition according to claim 1.

13. The aforementioned pharmaceutical composition is characterized by being a freeze-dried powder for injection. The pharmaceutical composition according to claim 1.

14. A method for producing a pharmaceutical composition according to any one of claims 1 to 13, (a1) Add the specified amount of antioxidant and glycine and mix uniformly, (a2) The solution obtained in step (a1) is characterized by comprising the step of adding a compound of formula I in the appropriate amount and dissolving it, Manufacturing method.

15. The manufacturing method according to claim 14, further comprising adding a pH adjusting agent in step (a1).

16. The manufacturing method according to claim 14, further comprising the step of (a3) ​​sterilizing and filtering the liquid obtained in step (a2) and freeze-drying it.

17. In step (a1) above, add the specified amount of antioxidant and glycine to the pH adjuster. In step (a2), the temperature of the solution obtained in step (a1) is controlled to 8 to 15°C, and the compound of formula I is further added in the appropriate amount. The manufacturing method according to claim 15.

18. The process is characterized by weighing the required amounts of sodium pyrosulfite and glycine, adding them to a sodium bicarbonate solution and stirring until completely dissolved, adjusting the pH to 7.0-9.0, controlling the solution temperature to 8-15°C, then adding the required amount of the compound of formula I, stirring for 30-50 minutes until dissolved, protecting the entire liquid formulation process with nitrogen gas, sterilizing and filtering, filling, simultaneously adding a bloomed butyl rubber stopper to an appropriate height, and freeze-drying. The manufacturing method according to claim 15.

19. Use of the pharmaceutical composition according to any one of claims 1 to 13 in the manufacture of a drug for preventing or treating liver failure.

20. The aforementioned liver failure is characterized by including acute liver failure, subacute liver failure, acute exacerbation of chronic liver failure, or chronic liver failure. The use described in claim 19.

21. The drug is further used to increase the survival rate of patients with liver failure and / or to improve liver function indicators in patients with liver failure, The use described in claim 20.

22. The improvement of the liver function indicators is characterized by including a decrease in alanine aminotransferase (ALT), a decrease in aspartate aminotransferase (AST), and / or a decrease in total bilirubin (TBil), The use described in claim 21.

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