Stable AST-3424 concentrated solution for injection

Abstract

< TableDetails num = '0001' > < Title / > < tbl: tgroup cool = '2' > < tbl: colspec cool = 'c001' cool width = '50%' / > < tbl: colspec cool width = 'c002' cool width = '50%' / > < tbl: cool width = '1' > impurity < / tbl: cool > < tbl: cool width = '1' > HPLC percentage content < / tbl The mobile phase B is a 8mmol / L ammonium acetate solution, and the solvent is a mixed solvent of acetonitrile and water in a volume ratio of 95: 5; and carrying out gradient elution at a flow rate of 1.0 ml / min.

Description

TECHNICAL FIELD

[0001] The present application relates to AST-3424 injection concentrated solution of patent application PCT / CN2020 / 101870, belongs to the field of medical research and development, especially relates to the quality control of injection concentrated solution. BACKGROUND

[0002] The DNA alkylating cancer treatment drug AST-3424 developed by our company takes overexpression of aldehyde ketone reductase 1C3 (AKR1C3) as a target (see patent application: DNA alkylating agent, corresponding PCT application number PCT / US2016 / 021581, publication number WO2016 / 145092, corresponding Chinese application number 2016800150788, publication number CN107530556A, which discloses compound TH2870; (R)- and (S)-1-(3-(3-N,N-dimethylaminocarbonyl)phenoxy-4-nitrophenyl)-1-ethyl-N,N'-bis(ethylene)aminophosphonate, compositions and methods of use and preparation thereof, corresponding PCT application number PCT / US2016 / 062114, publication number WO2017087428A1, corresponding Chinese application number 2016800446081, publication number CN108290911A, which discloses the S configuration compound), Chinese name (S)-1-(3-(3-N,N-dimethylaminocarbonyl)phenoxy-4-nitrophenyl)-1-ethyl-N,N'-bis(ethylene)aminophosphonate, also known as OBI-3424, TH-2870 S configuration compound), CAS number 2097713-69-2, the structure is as follows:

[0003]

[0004] Industry authority documents (Kathryn Evans, Jian Xin Duan, Tara Pritchard, et al. OBI-3424, a novel AKR1C3-activated prodrug, exhibits potent efficacy against preclinical models of T-ALL [J], Clinical Cancer Research, 2019, DOI: 10.1158 / 1078-0432.CCR-19-0551; Richard B. Lock, Kathryn Evans, Raymond Yung, Tara Pritchard, Beverly A. Teicher, Jian Xin Duan, Yue long Guo, Stephen W. Erickson and Malcolm A. Smith, Abstract LB-B16: The AKR1C3-Activated Prodrug OBI-3424 Exerts Profound In Vivo Efficacy Against Preclinical Models of T-Cell Acute Lymphoblastic Leukemia (T-ALL); a Pediatric Preclinical Testing Consortium Study [C], AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; October 26-30, 2017; Philadelphia, PA, DOI: 10.1158 / 1535-7163.) confirmed that the compound is a broad-spectrum small molecule anticancer prodrug, which has efficacy on various solid tumors and hematological tumors.

[0005] In order to carry out subsequent clinical trials, a large amount of AST-3424 bulk drug needs to be prepared to prepare formulations for long-term clinical trials (2-5 years).

[0006] Patent application PCT / CN2020 / 101870 discloses the prescription of AST-3424 injection preparation:

[0007] The mixed solvent is composed of 75% ethanol and 25% propylene glycol by volume, the AST-3424 content is 10 mg / ml, and the corresponding preparation process.

[0008] The formulations used in the subsequent clinical trials (CTR20191371, CTR20191399, CTR20201908, CTR20201915) are all the above formulations. SUMMARY

[0009] Since the clinical trials take a relatively long time, from phase I to the final phase III, which can be as long as 5 years or even longer, quality testing must be carried out during the post-production storage stage to ensure that the injection used in the clinical trials is safe and effective and meets the quality standards.

[0010] The inventors of the applicant team compared the results of storage experiments under multiple conditions, such as multiple batches, different temperatures, and whether to invert, and found that the different impurity content ranges under specific storage conditions can ensure that the quality of the above AST-3424 injection formulation meets the preset requirements to a certain extent, that is, by controlling the impurity profile (a series of impurities and their contents) of the above AST-3424 injection formulation, the quality of the AST-3424 injection formulation can be indirectly predicted or controlled, that is, the above impurity profile can become a marker for whether the quality of the AST-3424 injection formulation is qualified.

[0011] Specifically, the present application provides the following technical solutions.

[0012] A stable AST-3424 injection concentrate solution, the solvent of the solution is a mixture of ethanol and propylene glycol, and the volume ratio of ethanol to propylene glycol is 75:25, and the labeled content is 10 mg of AST-3424 raw material per 1 ml,

[0013] The injection concentrate solution is stored at -20±5℃ or lower temperature in a sealed light-proof condition, and after the sample is diluted with acetonitrile and detected by HPLC, the HPLC chemical purity of the AST-3424 raw material is greater than or equal to 98.0% after calculation by area normalization method, and the main signal peak of AST-3424 with an elution time of 11.0-15.0 min is taken as a control, and the impurities are represented by RRT, then the following impurities are contained:

[0014] Impurities HPLC percentage content RRT = 1.03-1.14 ≤0.14％ RRT = 1.14-1.24 ≤0.19％ RRT = 1.31-1.36 ≤0.21％ RRT = 1.37-1.39 ≤0.28％ RRT = 1.43-1.51 ≤0.21％ RRT = 1.46-1.55 ≤0.14％ RRT = 0.87-0.90 ≤0.14％

[0015] Among them,

[0016] The HPLC detection conditions are as follows:

[0017] UV detector, detection wavelength 230 nm,

[0018] The chromatographic column is Agilent Eclipse Plus C18, 150 mm x 4.6 mm, 3.5 μm,

[0019] Column temperature 25℃,

[0020] The mobile phase A is 10 mmol / L ammonium acetate solution, and the solvent is a mixed solvent of water and acetonitrile in a volume ratio of 95:5;

[0021] The mobile phase B is 8 mmol / L ammonium acetate solution, and the solvent is a mixed solvent of acetonitrile and water in a volume ratio of 95:5;

[0022] The flow rate is 1.0 ml / min for gradient elution, and the elution gradient is as follows:

[0023] Time (min) 0 8 15 30 35 35.1 45 Volume percentage of mobile phase B 20 30 30 90 90 20 20 Volume percentage of mobile phase A 80 70 70 10 10 80 80 .

[0024] Since there is ethanol and propylene glycol in the concentrated solution for injection of AST-3424, there is a relatively active alcohol hydroxyl group in the molecular structure of ethanol; and there is an amide group and an aziridine ring in AST-3424 which may decompose or react, therefore, in the subsequent long-term storage, high temperature (such as conventional 25℃, 5℃, etc.), high humidity, and light, contact with high molecular materials and their monomers in the packaging container, and environmental metal ions existing in the solution, all these conditions can cause new impurities in the storage process of AST-3424 injection.

[0025] Obviously, at any time after production is completed, the detected impurities can be divided into two categories:

[0026] The first type of impurity is the impurity brought in during production and preparation. These impurities exist after the production of AST-3424 bulk drug is completed, and they may be related to the process during the production of bulk drug, such as residues or by-products of raw materials, solvents, and reaction reagents. The content of these impurities does not change or changes slowly, and they can usually be reduced by changing or optimizing the bulk drug production process. Generally, their content does not change or changes slowly with the extension of storage time during subsequent storage.

[0027] The second type of impurity usually increases with the extension of storage time, especially it may grow slowly or not obviously at first, but it grows obviously after a certain period of time. This type of impurity is mostly a reactive impurity, that is, it may be the product of the reaction between the active groups such as amide group and aziridine ring in AST-3424 and ethanol, propylene glycol (including trace amounts of other alcohols, ethers, and aldehydes, and acids contained therein) during storage.

[0028] AST-3424 compound is purified by chromatography, and after purification, the AST-3424 chemical purity is detected by general non-chiral HPLC at 230 nm ultraviolet detector, and the area normalization method is used to calculate the detection result, which is greater than or equal to 98.0%, and the remaining 2.00% is various impurities. Here, impurities are in a narrow sense, only refer to the organic matter other than AST-3424 that can be detected under the above HPLC conditions, in fact, the above HPLC method cannot detect isomers AST-3423, water, residual solvents (such as can be detected in 230 nm ultraviolet monitor) and non-polar impurities and other substances in the injection solution.

[0029] When there is no control for the component to be tested (retention time tR1), another component in the sample or another known component added to the sample can be used as a reference (retention time tR2), and the relative retention time is used as a qualitative (or positioning) method. The relative retention time generally refers to the ratio of the retention time of the component to be tested to the retention time of the main component (RRT = tR1 / tR2).

[0030] The concentrated solution for injection is stored in a sealed light-proof environment at -20±5°C or lower temperature, and after dilution with acetonitrile, the sample is detected by HPLC, and after calculation by area normalization method, the HPLC chemical purity of AST-3424 drug substance is greater than or equal to 98.0%, and the main signal peak of AST-3424 with peak time of 11.0-15.0 min is used as a control, and the impurities are represented by relative peak time (also known as relative retention time) RRT, and the results are as follows:

[0031]

[0032] In fact, since the non-chiral chromatographic column used in general HPLC cannot separate AST-3424 from its isomer AST-3423, a part of the isomer cannot be completely separated in the detection by general non-chiral HPLC, therefore, the corresponding substances in the above 7 HPLC chromatographic peaks may be a single impurity, or a mixture of multiple impurities. But after detection of multiple batches and multiple conditions of AST-3424 concentrated solution for injection, after storage in a sealed light-proof environment at -20±5°C or lower temperature, the above HPLC method is used for detection, in addition to the main peak corresponding to AST-3424 drug substance, the 7 impurity peaks are completely present, therefore, it can be determined that the 7 HPLC chromatographic peaks correspond to 7 impurities or 7 groups of impurities.

[0033] In the subsequent continuous 24-month detection, it is found that the content of the 7 impurities or 7 groups of impurities changes slowly and almost unchanged throughout the time, and some are almost unchanged for a period of time and then the impurity content rises sharply at a certain time, so it can be concluded that the content of the 7 or 7 groups of impurities can be detected to control the quality of AST-3424 injection concentrate solution: that is, the AST-3424 injection concentrate solution product with impurity content within the above range can be determined as a qualified product, otherwise it is unqualified. Because for the impurities that change slowly and almost unchanged throughout the time, exceeding the above threshold range means that the first type of impurities in the batch product is too high, which means that the quality of the raw material drug is likely to be a problem; for the impurities that are almost unchanged for a period of time and then the impurity content rises sharply at a certain time, exceeding the above threshold value is likely to mean that the temperature is over-temperature during storage, the environment changes (light failure or closed environment is damaged), so even if the subsequent product detection may still meet the qualified standard, this batch of product must still be determined as a failed product and cannot be used for subsequent delivery as a clinical trial or commercialization.

[0034] The chromatographic column is Agilent Eclipse Plus C18, 150mmx4.6mm, 3.5μm, which is an Eclipse Plus series chromatographic column of Agilent Company in the United States, the filler is C18, the filler particle size is 3.5μm, the column body length is 150mm, and the inner diameter is 4.6mm.

[0035] Preferably, the injection concentrate solution is stored at-20±5℃ or lower temperature in a sealed light-proof condition, and the impurities are expressed by RRT as follows:

[0036]

[0037]

[0038] Preferably, the above AST-3424 injection concentrate solution is stored at-20±5℃ in a sealed light-proof condition, and the shelf life is greater than or equal to 24 months. The 24-month storage experimental data can undoubtedly prove that after 24 months of storage, each quality index in the raw material drug solution, especially the content of the above 7 or 7 groups of impurities meets the requirements, and no over-standard (exceeding the technical expectation) impurity or group of impurities is detected, so there is no risk of unqualified raw material drug quality or substandard storage conditions.

[0039] Applicant speculates that the shelf life of the above AST-3424 concentrated solution for injection is greater than or equal to 36 months based on the existing 24-month storage experiment results: comparison of different temperature storage experiments, the lower the temperature, the slower the impurity growth rate, so by further reducing the temperature, the storage time of the above AST-3424 concentrated solution for injection can be extended while the impurities and other quality indicators still meet the requirements (consistent with technical expectations).

[0040] Applicant speculates that the shelf life of the above AST-3424 concentrated solution for injection is greater than or equal to 48 months based on the existing 24-month storage experiment results: comparison of different temperature storage experiments, the lower the temperature, the slower the impurity growth rate, so by further reducing the temperature, the storage time of the above AST-3424 concentrated solution for injection can be extended while the impurities and other quality indicators still meet the requirements (consistent with technical expectations).

[0041] Applicant speculates that the shelf life of the above AST-3424 concentrated solution for injection is greater than or equal to 54 months based on the existing 24-month storage experiment results: comparison of different temperature storage experiments, the lower the temperature, the slower the impurity growth rate, so by further reducing the temperature, the storage time of the above AST-3424 concentrated solution for injection can be extended while the impurities and other quality indicators still meet the requirements (consistent with technical expectations).

[0042] Applicant speculates that the shelf life of the above AST-3424 concentrated solution for injection is greater than or equal to 54 months based on the existing 24-month storage experiment results: comparison of different temperature storage experiments, the lower the temperature, the slower the impurity growth rate, so by further reducing the temperature, the storage time of the above AST-3424 concentrated solution for injection can be extended while the impurities and other quality indicators still meet the requirements (consistent with technical expectations).

[0043] The stable AST-3424 concentrated solution for injection should meet the regulatory requirements for the mass limit of residual solvents in the solution.

[0044] The stable AST-3424 concentrated solution for injection does not add water, and the moisture content should be controlled during the corresponding production process, with a water content of 1.0% by mass.

[0045] Obviously, no water added means that no additional water is added during preparation, and other reagents, solvents, etc. used are anhydrous grade reagents, such as anhydrous ethanol, and water environment is avoided as much as possible or contact with water is avoided during production, that is, the water content needs to be controlled during the preparation process of the drug substance product of the present application. In theory, the lower the water content, the better the stability, but the research and development team considers the technical difficulty and the realization of industrial mass production, and the water content (determined by Karl Fischer method) is preferably within 1.0% by mass.

[0046] From the above, it can be seen that,

[0047] In particular, for the AST-3424 injection concentrate solution that has been determined, it can be determined by the product specification or other analytical detection methods that the injection contains AST-3424, and the solvent of the solution is a mixture of ethanol and propylene glycol, and the volume ratio of ethanol to propylene glycol is 75%:25%, and the labeled content is 10mg of AST-3424 drug substance per 1ml, which can be determined by the following fixed HPLC method:

[0048] UV detector, detection wavelength 230nm,

[0049] The chromatographic column is Agilent Eclipse Plus C18, 150mm x 4.6mm, 3.5μm,

[0050] Column temperature 25℃,

[0051] The mobile phase A is 10mmol / L ammonium acetate solution, and the solvent is a mixture of water and acetonitrile in a volume ratio of 95:5;

[0052] The mobile phase B is 8mmol / L ammonium acetate solution, and the solvent is a mixture of acetonitrile and water in a volume ratio of 95:5;

[0053] The flow rate is 1.0ml / min for gradient elution, and the elution gradient is as follows:

[0054] Time (min) 0 8 15 30 35 35.1 45 Volume percentage of mobile phase B 20 30 30 90 90 20 20 Volume percentage of mobile phase A 80 70 70 10 10 80 80

[0055] After detection, the area normalization method is used to detect and confirm that the following impurities are present, and the HPLC chemical purity of the AST-3424 drug substance is greater than or equal to 98.0% (as the main peak, it is used as a reference, and the peak time is 11.0-15.0min):

[0056] Impurities HPLC percentage content RRT = 1.03-1.14 ≤0.14％ RRT = 1.14-1.24 ≤0.19％ RRT = 1.31-1.36 ≤0.21％ RRT = 1.37-1.39 ≤0.28％ RRT = 1.43-1.51 ≤0.21％ RRT = 1.46-1.55 ≤0.14％ RRT = 0.87-0.90 ≤0.14％

[0057] It can be confirmed that the AST-3424 injection is the stable AST-3424 injection concentrate solution provided by the present application.

[0058] It is obvious that the stable AST-3424 injection concentrated solution provided above needs to be designed with a corresponding package during long-term storage, transportation or sale. Correspondingly, in order to keep it from deteriorating during storage, it needs to be packaged with a specially designed packaging unit.

[0059] The preparation packaging unit, as a basic unit for single administration, for clinical administration, comprises a container assembly, a label coated on the container assembly and a stable AST-3424 injection concentrated solution contained in the container,

[0060] The container assembly comprises a vial, a rubber stopper inserted into the vial mouth and a coating layer directly attached to the exposed surface of the rubber stopper facing the vial mouth and a closure cap for setting the vial mouth and the rubber stopper in engagement,

[0061] The material of the coating layer is selected from polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or tetrafluoroethylene / hexafluoropropylene copolymer FEP, and the closure cap is an aluminum-plastic composite cap.

[0062] The content recorded on the label includes:

[0063] The expiration date or shelf life is 24 months, 30 months, 36 months, 42 months, 48 months or 54 months, 60 months, 66 months, 72 months,

[0064] The storage condition is sealed, light-proof, -20±5℃ or lower temperature condition.

[0065] Experiments have proved that the use of polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or tetrafluoroethylene / hexafluoropropylene copolymer FEP as the material of the coating layer can ensure that no reaction occurs when it is directly in contact with the injection concentrated solution, thereby ensuring the stability of the injection concentrated solution.

[0066] Preferably, the coating layer is selected from tetrafluoroethylene / hexafluoropropylene copolymer FEP, and the rubber stopper is a chlorinated butyl rubber stopper.

[0067] In particular, unless otherwise specified, all numerical values in the specification have a range of ±10%, i.e. if a numerical range of 2-3 is specified in the specification, in fact, due to the existence of measurement error, accidental error, systematic error, the actual measurement result of 1.8-3.3 should also be considered to fall within the above range of 2-3, which is also in line with the requirements. BRIEF DESCRIPTION OF DRAWINGS

[0068] Figure 1 Figure 7 shows the trend of the change of 7 impurities with storage time in the stability test of the AST-3424 injection solution with batch number 219220301 stored at -20℃±5℃ and 5℃±3℃, in the light-proof and upright condition;

[0069] Figure 2 Figure 7 shows the trend of 7 impurities in AST-3424 injection solution with storage time in the storage stability test at 20℃±5℃ and 5℃±3℃, in the dark, upright storage.

[0070] Figure 3 Figure 7 shows the trend of 7 impurities in AST-3424 injection solution with storage time in the storage stability test at 20℃±5℃ and 5℃±3℃, in the dark, upright storage.

[0071] Figure 4 Figure 7 shows the trend of 7 impurities in AST-3424 injection solution with storage time in the storage stability test at 20℃±5℃ and 5℃±3℃, in the dark, upright storage.

[0072] Figure 5 Figure 7 shows the trend of 7 impurities in AST-3424 injection solution with storage time in the storage stability test at 20℃±5℃ and 5℃±3℃, in the dark, upright storage. DETAILED DESCRIPTION

[0073] The related impurity spectrum in the stable AST-3424 injection concentrated solution of the application is based on long-term stability data, and the following are specific experimental data.

[0074] General detection method

[0075] 1. HPLC detection method of AST-3424 injection concentrated solution

[0076] The HPLC detection conditions are as follows:

[0077] The HPLC detection conditions are as follows:

[0078] UV detector, detection wavelength 230nm,

[0079] The chromatographic column is Agilent Eclipse Plus C18, 150mm×4.6mm, 3.5μm,

[0080] Column temperature 25℃,

[0081] The mobile phase A is 10mmol / L ammonium acetate solution, and the solvent is a mixed solvent with a volume ratio of water:acetonitrile of 95:5;

[0082] The mobile phase B is 8mmol / L ammonium acetate solution, and the solvent is a mixed solvent with a volume ratio of acetonitrile:water of 95:5;

[0083] The flow rate is 1.0ml / min for gradient elution, and the elution gradient is as follows:

[0084] The flow rate is 1.0ml / min for gradient elution, and the elution gradient is as follows: Time (min) 0 8 15 30 35 35.1 45 Volume percentage of mobile phase B 20 30 30 90 90 20 20 Volume percentage of mobile phase A 80 70 70 10 10 80 80

[0085] Total run time is 45 minutes.

[0086] The HPLC instrument is an Agilent 1200 high performance liquid chromatograph equipped with a UV detector and an autosampler, or other equivalent instrument.

[0087] The sample is detected by HPLC after dilution with acetonitrile. The diluted solution is stable at 2-8°C for 3 days, and the detection should be completed within 3 days.

[0088] The injection volume is 5uL.

[0089] The labeled content of AST-3424 drug substance in the concentrated solution for injection (i.e. whether it is 10mg / ml) is detected and calculated using external standard method-single point correction method.

[0090] When detecting the impurity HPLC percentage content by injection, the related solution preparation requirements are:

[0091] Blank solution: acetonitrile.

[0092] Sensitivity solution: accurately measure the appropriate amount of reference solution, add acetonitrile to dilute to about 0.5ug AST-3424 per 1ml solution.

[0093] Reference solution (related substances control): accurately weigh the appropriate amount of AST-3424 control, add acetonitrile to dilute to about 0.02mg AST-3424 per 1ml solution, and prepare two copies in parallel. Use within 62 hours at 2-8°C.

[0094] Test solution: accurately measure the appropriate amount of the product, add acetonitrile to dilute to about 2mg AST-3424 per 1ml solution, as the test solution. Prepare two copies in parallel. Use within 53 hours at 2-8°C.

[0095] 2. System suitability evaluation of AST-3424 HPLC detection method and related parameter calculation

[0096] Before detecting the HPLC percentage content of each impurity in the HPLC solution, the HPLC system suitability evaluation should be performed and meet the requirements of the Chinese Pharmacopoeia 2020 edition.

[0097] Measurement and calculation

[0098] After the system suitability meets the requirements, the test solution is injected into the liquid chromatograph, and the chromatogram is recorded. With AST-3424 main component as control, external standard method-single point correction method is used to calculate the HPLC percentage content of impurities in the test solution.

[0099] The concentrated solution for injection in the present application was prepared according to the method described in patent application PCT / CN2020 / 101870, unless otherwise specified.

[0100] I. Preparation of AST-3424 concentrated solution for injection

[0101] Five batches were prepared according to the method described in patent application PCT / CN2020 / 101870:

[0102] Batch No. 219220301, the HPLC chemical purity of AST-3424 in the AST-3424 drug substance used was 99.70%,

[0103] Batch No. 219210701, the HPLC chemical purity of AST-3424 in the AST-3424 drug substance used was 99.50%,

[0104] Batch No. 20170701, the HPLC chemical purity of AST-3424 in the AST-3424 drug substance used was 99.60%,

[0105] Batch No. 20170702, the HPLC chemical purity of AST-3424 in the AST-3424 drug substance used was 99.50%,

[0106] Batch No. 20170703, the HPLC chemical purity of AST-3424 in the AST-3424 drug substance used was 99.40%.

[0107] II. Preparation of AST-3424 drug substance ethanol solution and long-term stability study

[0108] 2.1 Long-term stability study of different batches of AST-3424 drug substance ethanol combination under different storage temperatures, light and light

[0109] The three batches of 20170701, 20170702, and 20170703 concentrated solutions for injection were used to fill 1ml of injection solution in 2ml vials, and rubber stoppers were used for sealing. The accelerated condition was 5℃±3℃(2-8℃) and the long-term condition was -20℃±5℃. For the accelerated condition, samples were taken at 0M, 1M, 2M, 3M, and 6M, and for the long-term condition, samples were taken at 0M, 3M, 6M, 9M, 12M, 18M, 24M, 30M, 36M, 42M, 48M, 54M, 60M, 66M, and 72M for detection.

[0110] 219220301, 219210701 two batches of concentrated solution for injection using 7ml westlin bottle 1ml injection, and using rubber plug capping and sealing, to accelerate the condition: 5℃±3℃(2-8℃) and long-term condition:-20℃±5℃ storage, for accelerated condition at 0M, 1M, 3M and 6M sampling, long-term conditions 0M, 3M, 6M, 9M, 12M, 18M, 24M sampling detection.

[0111] The stored samples were temporarily stored at-20±5℃ until all tests were completed.

[0112] The HPLC percentage content of impurities was determined by "General Test Method Description-1, HPLC Test Method of AST-3424".

[0113] 2.2.1, 219220301 batch

[0114] The specific test results are as follows. In the table, if / is filled in, it means not applicable or below the detection limit, the same below.

[0115] Table 1: 219220301 batch number AST-3424 injection-20℃±5℃, light, upright storage stability test results statistics table

[0116]

[0117] Table 2: 219220301 batch number AST-3424 injection 5℃±3℃, light, upright storage stability test results statistics table

[0118]

[0119]

[0120] According to the data table of table 1, table 2, the corresponding appendix is made Figure 1 , record RRT=1.03~1.14 corresponding to impurity A, RRT=1.14~1.24 corresponding to impurity B, RRT=1.31~1.36 corresponding to impurity C, RRT=1.37~1.39 corresponding to impurity D, RRT=1.43~1.51 corresponding to impurity E, RRT=1.46~1.55 corresponding to impurity F, RRT=0.87~0.90 corresponding to impurity G.

[0121] Analysis of the stability data of 219220301 batch stored at different temperatures and known:

[0122] Comparing 5℃±3℃, -20℃±5℃, two storage temperatures, the lower the temperature, the more stable the storage.

[0123] Specifically, at -20℃±5℃, in the light, upright condition, only the six impurities ABCDEF were detected, and the G impurity was below the detection limit, with a small amount; moreover, with the extension of storage time, under the storage of 24 months, the impurities ABEF had almost no change or the content rising trend was not obvious, only the impurity CD slowly increased; accordingly, it can be inferred that the continued storage is still stable.

[0124] Specifically, at 5℃±3℃, in the light, upright condition, seven impurities ABCDEFG were detected, and moreover, with the extension of storage time, the rising trend of the impurity CDG content was very obvious, and ABEF had almost no change; by comparison, the growth rate of the impurity CD was faster. By comparing the impurity growth and total impurity, it can be known that the total impurity will exceed the standard after 6 months, and the impurities will be more and more in the subsequent storage, therefore, the product in this batch is stored at this temperature, and the maximum storage time is 1 month, and after 1 month, the impurity does not exceed the standard, but the impurity D has exceeded the standard (exceeding the technical expectation).

[0125] By comparing the impurity change under the above two temperature storage conditions, it can be known that the storage temperature of the above-mentioned concentrated solution for injection significantly affects the quality of the injection: high temperature will lead to an increase in impurities in the injection, and accelerate the rate of impurity generation.

[0126] 2.2.2, 219210701 batch

[0127] Table 3: 219210701 batch number AST-3424 injection -20℃±5℃, light, upright storage stability test result statistical table

[0128]

[0129] Table 4: 219210701 batch number AST-3424 injection 5℃±3℃, light, upright storage stability test result statistical table

[0130]

[0131]

[0132] According to the data table corresponding to table 3, table 4, the corresponding figures are made Figure 2 , RRT=1.03~1.14 corresponds to impurity A, RRT=1.14~1.24 corresponds to impurity B, RRT=1.31~1.36 corresponds to impurity C, RRT=1.37~1.39 corresponds to impurity D, RRT=1.43~1.51 corresponds to impurity E, RRT=1.46~1.55 corresponds to impurity F, and RRT=0.87~0.90 corresponds to impurity G.

[0133] The stability data of batch 219220701 stored at different temperatures are analyzed and it is found that:

[0134] Comparing the two storage temperatures of 5°C±3°C and -20°C±5°C, the lower the temperature, the more stable the storage.

[0135] Specifically, at -20°C±5°C, under light-protected and upright conditions, with the extension of storage time, the impurities ABEFG almost have no change or the rising trend is not obvious, and only the impurity CD slowly increases; accordingly, it can be inferred that the continued storage is still stable.

[0136] Specifically, at 5°C±3°C, under light-protected and upright conditions, the seven impurities ABCDEFG are detected, and with the extension of storage time, the rising trend of the impurities CDG content is very obvious, and ABEF almost has no change; in comparison, the growth rate of the impurity CD is faster. Comparing the impurity growth and total impurity, it can be found that the total impurity will exceed the standard after 6 months, and the impurities will be more and more in the subsequent storage, so the product of this batch is stored at this temperature, and the maximum storage time is 1 month, and after 1 month, the total impurity does not exceed the standard, but the impurities DE have exceeded the standard (exceeding the technical expectation).

[0137] Comparing the impurity change under the above two temperature storage conditions, it can be found that the storage temperature of the above-mentioned concentrated solution for injection directly determines the quality of the injection solution: high temperature will lead to an increase in impurities in the injection solution and accelerate the rate of impurity generation.

[0138] 2.2.3, batch 20170701

[0139] Table 5: 20170701 batch number AST-3424 injection solution -20°C±5°C, light-protected and upright storage stability test result statistical table

[0140]

[0141] Table 6: 20170701 batch number AST-3424 injection solution 5°C±3°C, light-protected and upright storage stability test result statistical table

[0142]

[0143]

[0144] According to the data tables of table 5 and table 6, the corresponding figures are made Figure 3, RRT = 1.03-1.14 corresponds to impurities for A, RRT = 1.14-1.24 corresponds to impurities for B, RRT = 1.31-1.36 corresponds to impurities for C, RRT = 1.37-1.39 corresponds to impurities D, RRT = 1.43-1.51 corresponds to impurities for E, RRT = 1.46-1.55 corresponds to impurities for F, RRT = 0.87-0.90 corresponds to impurities for G.

[0145] The stability data of batch 219220701 stored at different temperatures and can be known:

[0146] Comparing 5℃±3℃, -20℃±5℃, the lower the temperature, the more stable the storage.

[0147] Specifically, at -20℃±5℃, in the light, upright condition, impurity G is almost not detected; with the extension of storage time, under the storage of 48 months, the content of impurities ABCDEF has almost no change or the rising trend of content is not obvious, and after 48 months, impurities BCD have a slow growth trend.

[0148] Specifically, at 5℃±3℃, in the light, upright condition, 7 impurities ABCDEFG are detected, and with the extension of storage time, the rising trend of impurity CD content is very obvious, and ABEFG has almost no change. Comparing the impurity growth and total impurity, it can be known that the total impurity will exceed the standard after 6 months, and the impurities will be more and more in subsequent storage, so the product in this batch is stored at this temperature, and the maximum storage time is 6 months. After 6 months, the total impurity does not exceed the standard, but impurity D has exceeded the standard (exceeding the technical expectation).

[0149] Comparing the impurity change under the above two temperature storage conditions, it can be known that the storage temperature of the above-mentioned concentrated solution for injection directly determines the quality of the injection solution: high temperature will lead to an increase in impurities in the injection solution and accelerate the rate of impurity generation.

[0150] 2.2.4, 20170702 batch

[0151] Table 7: 20170702 batch number AST-3424 injection-20℃±5℃, light, upright storage stability test result statistical table

[0152]

[0153] Table 8: 20170702 batch number AST-3424 injection 5℃±3℃, light, upright storage stability test result statistical table

[0154]

[0155] According to the data table corresponding to table 7, table 8, the corresponding appendix is madeFigure 4 , record RRT = 1.03 ~ 1.12 corresponding to impurities A, RRT = 1.14 ~ 1.23 corresponding to impurities B, RRT = 1.31 ~ 1.36 corresponding to impurities C, RRT = 1.37 ~ 1.39 corresponding to impurities D, RRT = 1.43 ~ 1.51 corresponding to impurities E, RRT = 1.46 ~ 1.55 corresponding to impurities F, RRT = 0.87 ~ 0.90 corresponding to impurities G.

[0156] The stability data of batch 219220701 stored at different temperatures and can be known:

[0157] Comparing 5°C ± 3°C, -20°C ± 5°C, the lower the temperature, the more stable the storage.

[0158] Specifically, at -20°C ± 5°C, in the dark, upright condition, impurity G was almost not detected; with the extension of storage time, the content of impurities ABCDEF almost did not change or the content rising trend was not obvious.

[0159] Specifically, at 5°C ± 3°C, in the dark, upright condition, 7 impurities ABCDEFG were detected, and with the extension of storage time, the rising trend of impurity CD content was very obvious, and ABEFG almost did not change. By comparing the impurity growth and total impurity, it can be seen that the total impurity will exceed the standard after 6 months, and the impurity will increase more and more in subsequent storage, so the product in this batch is stored at this temperature. The maximum storage time is 6 months, and after 6 months, the total impurity does not exceed the standard, but the impurity D has exceeded the standard (exceeding the technical expectation).

[0160] By comparing the impurity change under the above two temperature storage conditions, it can be seen that the storage temperature of the above injection solution directly determines the quality of the injection solution: high temperature will lead to an increase in impurities in the injection solution and accelerate the rate of impurity generation.

[0161] 2.2.5, 20170703 batch

[0162] Table 9: 20170703 batch number AST-3424 injection solution -20°C ± 5°C, light, upright storage stability test result statistical table

[0163]

[0164]

[0165] Table 10: 20170703 batch number AST-3424 injection solution 5°C ± 3°C, light, upright storage stability test result statistical table

[0166]

[0167] According to the corresponding data table of Table 9, Table 10, the corresponding appendix is made Figure 5 , record RRT = 1.03 ~ 1.12 corresponding to impurities A, RRT = 1.14 ~ 1.23 corresponding to impurities B, RRT = 1.31 ~ 1.36 corresponding to impurities C, RRT = 1.37 ~ 1.39 corresponding to impurities D, RRT = 1.43 ~ 1.51 corresponding to impurities E, RRT = 1.46 ~ 1.55 corresponding to impurities F, RRT = 0.87 ~ 0.90 corresponding to impurities G.

[0168] Analysis of the stability data of 219220701 batch stored at different temperatures and can be known:

[0169] Comparing 5℃±3℃, -20℃±5℃, two storage temperatures, the lower the temperature, the more stable the storage.

[0170] Specifically, at -20℃±5℃, light, upright conditions, impurity G was almost not detected; with the extension of storage time, the content of impurities ABCDEF was almost unchanged or the rising trend of content was not obvious.

[0171] Specifically, at 5℃±3℃, light, upright conditions, 7 impurities ABCDEFG were detected, and with the extension of storage time, the rising trend of impurity CD content was very obvious, and AB EFG was almost unchanged. By comparing the impurity growth and total impurity, it can be seen that the total impurity of impurity D has exceeded the standard at 6 months, and the subsequent storage impurities will be more and more, therefore, the product of this batch is stored at this temperature, the maximum storage time is 6 months, after 6 months, the total impurity has not exceeded the standard, but impurity D has exceeded the standard (exceeding the technical expectation).

[0172] By comparing the impurity change under the above two temperature storage conditions, it can be seen that the storage temperature of the above injection concentrated solution directly determines the quality of the injection: high temperature will lead to an increase in impurities in the injection and accelerate the rate of impurity generation.

Claims

1. A stable AST-3424 injection concentrated solution, the solvent of the solution is a mixture of ethanol and propylene glycol, and the volume ratio of ethanol and propylene glycol is 75%:25%, and the labeled content is 10 mg of AST-3424 active pharmaceutical ingredient per 1 ml, The concentrated solution is stored at -20±5℃ or lower temperature in a sealed and light-protected condition, after the sample is diluted with acetonitrile, the HPLC detection is performed, and after the area normalization calculation, the HPLC chemical purity of the AST-3424 active pharmaceutical ingredient is greater than or equal to 98.0%, the main signal peak of AST-3424 with an elution time of 11.0-15.0 min is taken as a control, and the impurities are represented by RRT, and the impurities include: wherein, The HPLC detection conditions are as follows: an ultraviolet detector, a detection wavelength of 230 nm, a chromatographic column of Agilent Eclipse Plus C18, 150 mm×4.6 mm, 3.5 μm, a column temperature of 25℃, a mobile phase A of 10 mmol / L ammonium acetate solution, a solvent of a mixed solvent with a volume ratio of water:acetonitrile of 95:5; a mobile phase B of 8 mmol / L ammonium acetate solution, a solvent of a mixed solvent with a volume ratio of acetonitrile:water of 95:5; a gradient elution at a flow rate of 1.0 ml / min, and the elution gradient is as follows: 。 2. The stable AST-3424 injection concentrated solution according to claim 1, the concentrated solution is stored at -20±5℃ or lower temperature in a sealed and light-protected condition, and the impurities are represented by RRT, and the impurities include: 。 3. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 24 months.

4. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 30 months.

5. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 36 months.

6. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 42 months.

7. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 48 months.

8. The stable AST-3424 injection concentrated solution according to claim 1, which is stored at -20±5℃ in a sealed and light-protected condition, and has a shelf life of greater than or equal to 54, 60, 66 or 72 months.

9. A formulation packaging unit as a basic unit for single administration for clinical administration, comprising a container assembly, a label covering the container assembly, and the stable AST-3424 injection concentrated solution according to any one of claims 1-8 contained in the container, The container assembly comprises a vial, a rubber stopper inserted into the vial opening, and a coating layer directly attached to the exposed surface of the rubber stopper facing the vial opening and a cap for setting on the rubber stopper to snap-fix the vial opening and the rubber stopper, The material of the coating layer is selected from polytetrafluoroethylene PTFE, polyvinylidene fluoride PVDF or tetrafluoroethylene / hexafluoropropylene copolymer FEP, The content recorded on the label includes: The expiration date or shelf life is 24 months, 30 months, 36 months, 42 months, 48 months or 54 months, 60 months, 66 months, 72 months, The storage condition is sealed, light-proof, -20±5℃ or lower temperature condition.

10. The formulation packaging unit according to claim 9, wherein the coating layer is selected from tetrafluoroethylene / hexafluoropropylene copolymer FEP, and the rubber stopper is a chlorinated butyl rubber stopper.

Images