Dipeptide derivative compositions, and methods for their preparation 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 priority to a prior application bearing patent application number 202210532274.X and entitled "Dipeptide Derivative Composition, and Its Preparation Method and Use," filed with the State Intellectual Property Office of China on May 6, 2022, the entirety of which is incorporated herein by reference.

[0002] [Technical field] The present invention relates to the field of pharmaceutical formulations, specifically to a dipeptide derivative composition, and its preparation and use.

[0003] [Background technology] Liver failure refers to severe liver damage caused by multiple factors, which causes severe impairment or decompensation in the liver's own functions such as synthesis, detoxification, excretion and biotransformation, and clinically manifests as a group of syndromes represented by disorders of the thrombin mechanism, jaundice, hepatic encephalopathy, dehydration, etc. The mortality rate of liver failure is extremely high.

[0004] Patent applications CN201110025509.8 and CN201110025516.8 disclose dipeptide derivatives that can be used to treat liver failure, the structures of which are as follows:

[0005] [ka]

[0006] The chemical name of this dipeptide derivative is 3-(2-benzyloxycarbonylamino-3-methyl-butanamido)-5-fluoro-4-oxo-pentanoic acid (F573), which can significantly inhibit or reverse liver failure, has significant therapeutic effect on liver failure, and has no obvious toxicity to cells.

[0007] In the oral and gastrointestinal environments, the dipeptide derivative is easily destroyed by various enzymes in the oral and gastrointestinal tract, and is also easily lost in efficacy due to the first-pass effect of the liver. Therefore, it is not suitable for designing an oral dosage form. Furthermore, research has found that the dipeptide derivative has poor stability in aqueous solution and cannot withstand moist heat sterilization, so there is a need to develop an injection dosage form with high stability, low impurity content, and low side effects.

[0008] Summary of the Invention In order to remedy the problems existing in the prior art, in a first aspect, the present invention provides a method for producing ... liquid crystal display comprising: (a) a compound of formula I, having the structure:

[0009] [ka]

[0010] , (b) glycine, (c) an antioxidant.

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

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

[0013] According to an embodiment of the present invention, the 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%, 10.0% (w / w). Preferably, the composition (a) accounts for about 2.0% to about 5.0% (w / w) of the total amount of the composition, According to an embodiment of the present invention, the composition (b) is 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%, 15.0% (w / w). Preferably, the composition (b) is about 3.0% to about 10.0% (w / w) of the total amount of the composition, According to an embodiment of the present invention, the 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%, 0.50% (w / w). Preferably, the composition (c) accounts for about 0.05% to about 0.30% (w / w) of the total amount of the composition, According to an embodiment of the present invention, the pH value of the composition is greater than 7.0, and in some embodiments, the pH value of the composition is in the range of 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, 9.0, and preferably 7.5 to 8.5.

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

[0015] In a second aspect, the present invention provides a method for producing a method for the treatment of a pulmonary arthritis, comprising: (a1) adding the antioxidant, glycine, and optionally a pH adjuster in the prescribed amounts, and mixing uniformly; (a2) adding and dissolving an amount of a compound of formula I in the solution obtained in step (a1); The present invention also provides a method for producing the pharmaceutical composition, comprising:

[0016] According to an embodiment of the present invention, a method for producing the pharmaceutical composition includes the steps of: (a3) filtering and sterilizing the liquid obtained in step (a2), followed by freeze-drying.

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

[0018] According to an embodiment of the present invention, in the step (a3), a Millipore filter membrane is used for sterilization filtration, and preferably, the Millipore filter membrane is a polyethersulfone Millipore filter membrane.

[0019] According to a preferred embodiment of the present invention, the method for preparing the pharmaceutical composition includes: The process includes weighing out the amount of sodium pyrosulfite and glycine, adding them to the sodium bicarbonate solution and stirring to completely dissolve them, adjusting the pH value to 7.0-9.0, controlling the solution temperature to 8-15°C, then adding the amount of F573, stirring for 30-50 minutes to dissolve them, protecting the entire liquid blending process with nitrogen gas, sterilizing and filtering, filling, and simultaneously adding butyl rubber stoppers in the open state to an appropriate height, and then freeze-drying. Preferably, the process further includes the steps of box pressure stoppering, box removal, capping, lamp inspection, labeling, packaging, sample submission, and warehousing of the finished product after passing the inspection after freeze-drying.

[0020] In a third aspect, the present invention provides the use of the pharmaceutical composition described above in the manufacture of a medicament for the prevention or treatment of liver failure.

[0021] According to an embodiment of the present invention, the liver failure includes acute liver failure, subacute liver failure, and acute exacerbation of chronic liver failure and chronic liver failure.

[0022] According to an embodiment of the present invention, the medicament is further used to improve the survival rate of patients with liver failure and / or improve liver function indicators of patients with liver failure. In some embodiments, improving the liver function indicators includes reducing alanine aminotransferase (ALT), reducing aspartate aminotransferase (AST), and / or reducing total bilirubin (TBil).

[0023] (Beneficial Effects) The present invention provides a stable composition of a dipeptide derivative of formula I, which has good compatibility with auxiliary agents, and the lyophilized preparation prepared based on the composition has a low impurity content and good stability under high humidity, strong light and low temperature storage conditions.

[0024] [Mode for carrying out the invention] The technical solution of the present invention will be described in more detail below with reference to specific examples. It should be understood that the following examples are merely illustrative and interpreted for the present invention, and should not be interpreted as limiting the scope of the claims of the present invention. Any technology realized based on the above content of the present invention is included in the scope of the claims of the present invention.

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

[0026] reagent 3-(2-benzyloxycarbonylamino-3-methyl-butanamido)-5-fluoro-4-oxo-pentanoic acid (abbreviated as F573, provided by Beijing ContiNic Pharmaceutical Co., Ltd.), the structure is as follows:

[0027] [ka]

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

[0029] Example 1: Preparation of F573 injection 1.1 Screening of injection solvents for the purpose of producing an injection solution of F573.

[0030] [Table 1]

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

[0032] 1.2 Stability considerations for F573 injection F573 was dissolved in a propylene glycol / water solution (80% / 20%, pH adjusted to 4.0-5.0), sterilized at high temperature for 15 minutes at 120°C for 15 minutes, and then tested (see Table 1-2 below for the results).

[0033] [Table 2]

[0034] The results show that in the samples after high-temperature sterilization, the content of related substances increased by about 50%, and the content of F573 decreased by about 50%. F573 has poor stability in aqueous solution and cannot withstand moist heat sterilization, which means that it is not suitable for development into small-volume or large-volume injections.

[0035] (Example 2: Powdered injection (preparation using F573 + non-aqueous solvent)) To produce F573 powder injection in a composite package, each set contains one bottle of sterile powder containing 30 mg of active ingredient and 2 mL of non-aqueous solvent, which is prepared immediately and used immediately, and an appropriate amount of analgesic (benzyl alcohol) is added appropriately. First, 90% ethanol solution is used as the solvent, and the F573 raw material is dissolved, crystallized, decarbonized, dried, etc. are performed. The results show that the yield is about 50% by performing the recrystallization and decarbonization processes on the raw material, and problems such as changes in the crystal form may occur. Therefore, F573 is not suitable for development into the above dosage form.

[0036] Example 3 Lyophilized Composition Formulation Study 3.1 Experiments on the effect of adjuvants In freeze-dried preparations, mannitol can be used as a carrier to form a uniform skeleton, while amino acids can be used as a skeleton agent for freeze-dried agents, and are also common protein protectants. The formulation is selected by adding no other auxiliary agents to F573, and adding arginine, glycine, and mannitol, respectively, and then sieving F573 through a 100 mesh sieve to prepare for use. The freeze-dried process is adopted to manufacture the product. An appropriate amount of sodium bicarbonate is weighed out and prepared into a 1 mol / L solution to prepare for use. The blended amount of arginine, glycine, or mannitol is weighed out, added to the above sodium bicarbonate solution and stirred to completely dissolve, the solution temperature is controlled to 8-15°C, and then the blended amount of F573 is added, stirred while adding, and stirred for 40 minutes to dissolve, and nitrogen gas protection is performed during the entire liquid blending process. The terminal filter was a 0.22 μm polyethersulfone Millipore filter membrane (filter cartridge) for sterilization filtration, filling, and at the same time adding a butyl rubber stopper in a bloomed state to an appropriate height, followed by freeze-drying to obtain a lump.

[0037] Research results show that glycine is a more suitable backbone agent, as shown in Table B-1 below.

[0038] [Table 3]

[0039] 3.2 Based on the results of the compatibility test between F573 and auxiliary agents, research was conducted on the combination of F573 with glycine and sodium pyrosulfite, as shown in the following Tables B-2 and B-3, which show that F573 is suitable for combination with glycine and sodium pyrosulfite.

[0040] [Table 4]

[0041] [Table 5]

[0042] 3.3 Mixture optimization experiment Sodium pyrosulfite and glycine were selected as the auxiliary agents for the F573 lyophilized formulation, and the formulation was further optimized by considering these two factors. Each factor was set to three levels, and nine formulations were arranged using the orthogonal array L9 (34) to conduct experiments. The physical appearance of the lyophilized needle, reconstitution time and pH value were used as evaluation indexes to analyze the results. 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: Lyophilized preparation F-1 4.1 F-1 Production Formulation F-1 (1000 units) F573 30g Glycine 50g Sodium pyrosulfite 1 g Sodium bicarbonate (appropriate amount) Water for injection was added to 1000 mL.

[0047] F573 is sieved through a 100 mesh sieve and prepared for use. An appropriate amount of sodium bicarbonate is weighed out to prepare a 1 mol / L solution and prepared for use. Separately, the blended amount of sodium pyrosulfite and glycine are weighed out, added to the above sodium bicarbonate solution and stirred to completely dissolve, the pH value is adjusted to 7.5-8.5, the solution temperature is controlled at 8-15°C, and then the blended amount of F573 is added, stirred while being added, and stirred for 40 minutes to dissolve, and nitrogen gas protection is performed during the entire liquid blending process. After inspection, pH and content pass, it is filtered. The terminal filter is adopted to sterilize and filter, and after the visible foreign matter inspection of the medicinal liquid is passed, it is placed in a pharmaceutical large bottle, and after adjusting the amount of charge, it is filled, and at the same time, a butyl rubber stopper in the open state is added to the appropriate height. Freeze drying, box pressure stoppering, box removal, capping, lamp inspection, labeling, packaging, sample submission, and storing the finished products after they pass inspection. The finished products were stored at 4-8℃.

[0048] 4.2 Three batches of samples were taken according to the above F-1 manufacturing process, and the long-term stability of the preparations under high humidity, strong light and low temperature storage conditions was examined, and the test results are shown below. The results show that all the test items of F573 for injection are in accordance with the pharmacopoeia's requirements for the stability of preparations.

[0049] [Table 9]

[0050] [Table 10]

[0051] [Table 11]

[0052] [Table 12]

[0053] [Table 13]

[0054] [Table 14]

[0055] The exemplary embodiments of the present invention have been described above. However, the scope of the claims of the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the scope of the spirit and principles of the present invention should be included within the scope of the claims of the present invention.

Claims

1. 1. A pharmaceutical composition comprising: (a) a compound of formula I, having the structure: 【Chemistry 1】 、 (b) glycine, (c) an antioxidant; Pharmaceutical compositions.

2. The antioxidant is one, two or more selected from the group consisting of ascorbic acid, sodium edetate, sodium hydrogen sulfite and sodium pyrosulfite. The pharmaceutical composition of claim 1.

3. The pharmaceutical composition further comprises a component (d) a pH adjuster. The pharmaceutical composition of claim 1.

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

5. The composition (a) accounts for 0.5% to 10.0% (w / w) of the total amount of the pharmaceutical composition; said composition (b) comprising about 0.5% to about 15.0% (w / w) of the total amount of said pharmaceutical composition; The composition (c) accounts for 0.01% to 0.50% (w / w) of the total amount of the pharmaceutical composition; The pharmaceutical composition of claim 1.

6. The composition (a) accounts for 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) accounts for 3.0% to about 10.0% (w / w) of the total amount of the pharmaceutical composition. The pharmaceutical composition according to claim 5.

8. The 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 pharmaceutical composition has a pH value greater than 7.

0. The pharmaceutical composition of claim 1.

10. The pharmaceutical composition has a pH value of 7.0 to 9.

0. The pharmaceutical composition of claim 9.

11. The pharmaceutical composition has a pH value of 7.5 to 8.

5. The pharmaceutical composition of claim 9.

12. Further, water for injection is contained in an amount to bring the total volume to 1000 mL, The composition (a) contains 30 g, the composition (b) contains 50 g, and the composition (c) contains 1 g, and The pharmaceutical composition is characterized in that its pH value is 7.5 to 8.

5. The pharmaceutical composition of claim 1.

13. The pharmaceutical composition is a freeze-dried powder for injection. The pharmaceutical composition of claim 1.

14. A method for producing the pharmaceutical composition according to any one of claims 1 to 13, comprising: (a1) adding the antioxidant and glycine in the appropriate amounts and mixing uniformly; (a2) adding and dissolving the compound of formula I in the solution obtained in step (a1); Manufacturing method.

15. The manufacturing method described in Claim 14, characterized in that in step (a1), it further comprises adding a pH adjuster.

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

17. In the step (a1), the antioxidant and glycine are added in the amount required to adjust the pH, In the step (a2), the temperature of the solution obtained in the step (a1) is controlled to 8 to 15°C, and a compound of formula I is further added. The method of claim 15.

18. The method comprises the steps of: weighing out the required amount of sodium pyrosulfite and glycine, adding them to a sodium bicarbonate solution and stirring to completely dissolve them; adjusting the pH to 7.0-9.0; controlling the solution temperature at 8-15°C; adding the required amount of the compound of formula I; stirring for 30-50 minutes to dissolve them; protecting the entire liquid blending process with nitrogen gas; sterilizing and filtering; filling; simultaneously adding bloomed butyl rubber stoppers to an appropriate height; and freeze-drying. The method of claim 15.

19. Use of the pharmaceutical composition of any one of claims 1 to 13 in the manufacture of a medicament for the prevention or treatment of liver failure.

20. The liver failure includes acute liver failure, subacute liver failure, acute exacerbation of chronic liver failure, or chronic liver failure.

20. The use according to 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.

21. The use according to claim 20.

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

22. The use according to claim 21.

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