4-acetyl-5-phenylpyrrolidine-2,3-dione compounds and their use as cdk2 protein inhibitors
By providing 4-acetyl-5-phenylpyrrolidine-2,3-dione compounds as CDK2 protein inhibitors, the problem of lack of CDK2 target drugs in the existing technology is solved, and effective inhibition and treatment of CDK2 protein-related diseases are achieved.
Patent Information
- Application Number
- CN202011398331.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2020-12-04
- Publication Date
- 2025-10-10
- Estimated Expiration
- 2040-12-04
AI Technical Summary
Currently, there are no marketed drugs targeting CDK2 protein, and there is a lack of effective CDK2 protein inhibitors for cancer treatment.
Provided are 4-acetyl-5-phenylpyrrolidine-2,3-dione compounds or pharmaceutically acceptable salts thereof as CDK2 protein inhibitors for use in preparing drugs for preventing and treating diseases targeting CDK2 protein. The dosage forms include oral and injectable dosage forms.
Effectively inhibiting the activity of CDK2 protein provides important research value for the preparation of drugs to treat diseases targeting CDK2 protein, and achieves the prevention and treatment of related diseases.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of medicine, and relates to a 4-acetyl-5-phenyl pyrrolidine-2,3-dione compound and application thereof in CDK2 protein inhibitors. BACKGROUND
[0002] Cyclin-dependent kinases (CDKs) are a family of serine / threonine protein kinases with 13 members, including CDK1-CDK13. CDKs, in concert with cyclins, are important factors in the regulation of cell cycle. The cell cycle can only do the right thing at the right time depending on their ordered activation. Therefore, the synergistic effect of CDK and cyclin is an important factor in the regulation of cell cycle. According to the different functions of CDK, CDKs can be divided into two categories: one is involved in cell cycle regulation, and the other is involved in transcriptional regulation. In recent years, the clinical development of CDK4 / 6 inhibitors has changed the treatment of breast cancer, stimulated interest in this field, and increased the research efforts on other members of the CDK family as therapeutic targets.
[0003] Cyclin-dependent kinase 2 is an important member of the CDK family. In dividing cells, CDK2 is a core cell cycle regulator that is active from late G1 to the entire S phase. The wide range of functions of CDK2 in cell proliferation and survival pathways makes it an ideal target for mechanism-based and low-toxicity therapeutic strategies in cancer treatment. There is a large amount of evidence that the activity of CDK2 also affects cell differentiation and adaptive immune responses. CDK2 is one of the most potential targets in the CDK family. However, there is still no marketed drug targeting CDK2 protein so far. SUMMARY
[0004] In order to solve the problems existing in the prior art, the purpose of the present application is to provide a 4-acetyl-5-phenyl pyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof and application thereof as a CDK2 protein inhibitor, which can effectively inhibit the activity of CDK2 protein.
[0005] The present application provides a 4-acetyl-5-phenyl pyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof, the skeleton of the compound is shown in the following formula (I):
[0006]
[0007] wherein,
[0008] R 11 is H, C 1-10 alkyl, C 1-10 alkoxy, i-Pr, halogen, one or more of H, halogen, C
[0009] R 12 is H, halogen, C 1-10 alkyl, alkoxy and the like;
[0010] R 13 is H, halogen, C 1-10 alkyl, alkoxy and the like;
[0011] R 14 is H, C 1-10 alkyl, alkoxy and the like;
[0012] R2is Ph, C 1-10 alkyl, alkoxy and the like;
[0013] R3is C 1-10 alkyl, alkoxy and the like;
[0014] Preferably,
[0015] R 11 is H-, Et-, MeO-, EtO-, i-Pr, F-, Cl-, t-Bu, n-BuO and the like;
[0016] R 12 is H-, F-, MeO- and the like;
[0017] R 13 is H-, F-, Cl, -MeO- and the like;
[0018] R 14 is H, MeO- and the like;
[0019] R2is Ph-,
[0020] R3is
[0021] The present application also provides the use of the above-mentioned 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof as a CDK2 protein inhibitor.
[0022] The present application also provides the use of the above-mentioned 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and / or treating a disease targeting CDK2 protein.
[0023] The present application also provides a pharmaceutical composition comprising the 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or its pharmaceutically acceptable salt as shown in formula (I) and a pharmaceutically acceptable carrier.
[0024] The present application also provides the use of the above-mentioned pharmaceutical composition as a CDK2 protein inhibitor.
[0025] The present application also provides the use of the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating diseases targeting CDK2 protein.
[0026] The dosage form of the pharmaceutical composition is an oral dosage form or an injection dosage form.
[0027] The oral dosage form includes tablets, capsules, films, granules, etc., and also includes sustained-release or non-sustained-release dosage forms, etc.
[0028] The present application also provides a method for preventing and / or treating related diseases, which comprises administering the 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or its pharmaceutically acceptable salt or the pharmaceutical composition as described above to a subject in need thereof.
[0029] The dosage form of the pharmaceutical composition is an oral dosage form or an injection dosage form.
[0030] The oral dosage form includes tablets, capsules, films, granules, etc., and also includes sustained-release or non-sustained-release dosage forms, etc.
[0031] The beneficial effects of the present application include that the 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or its pharmaceutically acceptable salt provided by the present application can be used as a CDK2 protein inhibitor, can effectively inhibit the activity of CDK2 protein, and provides important research value for the preparation of a medicament for treating diseases targeting CDK2 protein. DETAILED DESCRIPTION
[0032] The present application is further described in detail in combination with the following specific examples. The process, conditions, experimental methods, etc. for implementing the present application are the general knowledge and common sense in the art, and the present application does not have special limitations.
[0033] Application Examples
[0034] I. Experimental materials:
[0035] CDK2 (eurofins, Cat.No.14-448M, Lot.No.D7NN039U-G)
[0036] Substrate Peptide FAM-P18 (GL Biochem, Cat. No. 114202, Lot. No. P080319-XY114202)
[0037] ATP (Sigma, Cat. No. A7699-1G, CAS No. 987-65-5)
[0038] DMSO (Sigma, Cat. No. D2650, Lot. No. 474382)
[0039] EDTA (Sigma, Cat. No. E5134, CAS No. 60-00-4)
[0040] 96-Well Plates (Corning, Cat. No. 3365, Lot. No. 22008026)
[0041] 384-Well Plates (Corning, Cat. No. 3573, Lot. No. 12608008)
[0042] Positive Control Staurosporine (Selleckchem, Cat. No. S1421, Lot. No. S142105)
[0043] II. Experimental Methods:
[0044] 1. Prepare 1X Kinase Base Buffer and Stop Buffer
[0045] 1) 1X Kinase Base Buffer
[0046] 50 mM HEPES, pH 7.5
[0047] 0.0015% Brij-35
[0048] 10 mM MgCl2
[0049] 2 mM DTT
[0050] 2) Stop Buffer Solution
[0051] 100 mM HEPES, pH 7.5
[0052] 0.015% Brij-35
[0053] 0.2% Coating Reagent #3
[0054] 50 mM EDTA
[0055] 2. Prepare Compounds
[0056] 1) Dilute 4-acetyl-5-phenylpyrrolidine-2,3-diones (see Table 1) shown in Formula (I) in 100% DMSO to 50 times the final desired highest inhibitor concentration in the reaction. Then transfer 100 μl of the compound dilution to a well of a 96 well plate. For example, if the desired highest inhibitor concentration is 100 μM, then prepare a 5000 μM compound solution in DMSO at this step.
[0057] 2) In the same 96 well plate, add 100 μl of 100% DMSO to two empty wells as a no compound added control and a no enzyme added control. Label this plate as the source plate.
[0058] 3) Prepare intermediate plate: transfer 10 μl of compound from the source plate to a new 96 well plate as the intermediate plate; add 90 μl of 1x kinase buffer to each well of the intermediate plate; mix the compound on the intermediate plate on a shaker for 10 minutes.
[0059] 3. Prepare assay plate
[0060] Transfer 5 μl per well from the 96 well intermediate plate to a 384 well plate in duplicate. For example, transfer A1 of the 96 well plate to A1 of the 384 well plate. Transfer A2 of the 96 well plate to A3 of the 384 well plate, and so on.
[0061] 4. Enzyme reaction
[0062] 1) Prepare 2.5x enzyme solution: add kinase to 1x kinase base buffer.
[0063] 2) Prepare 2.5x peptide solution: add FAM labeled peptide and ATP to 1x kinase base buffer.
[0064] 3) The assay plate already contains 5 μl of compound in a 10% DMSO solution.
[0065] 4) Transfer 2.5x enzyme solution to the assay plate, add 10 μl of 2.5x enzyme solution to each well of the 384 well assay plate.
[0066] 5) Incubate at room temperature for 10 minutes.
[0067] 6) Transfer 2.5x peptide solution to the assay plate, add 10 μl of 2.5x peptide solution to each well of the 384 well assay plate.
[0068] 7) Kinase reaction and stop: incubate at 28°C for the specified time, 30 min, add 25 μl of stop buffer to stop the reaction.
[0069] 5. Enzyme reader readout
[0070] Data were collected on a microplate reader.
[0071] 6. Curve Fitting
[0072] 1) Copy the conversion data from the Caliper program.
[0073] 2) Convert the reading value to an inhibition value.
[0074] Inhibition rate % = DMSO control - experimental group) / (DMSO control - blank control) * 100.
[0075] 3) Data were fitted in XLFit excel add-in version 5.4.0.8 to obtain IC50 values.
[0076] 3. The results are shown in Table 1:
[0077] Table 1
[0078]
[0079]
[0080]
[0081] Note: Biological activity: >100,000 nM is poor; 50,000-100,000 nM is moderate; 20,000-50,000 nM is good; 1,000-20,000 nM is excellent; <1,000 is optimal
[0082] The data in Table 1 indicate that the 4-acetyl-5-phenylpyrrolidine-2,3-dione compounds of formula (I) or pharmaceutically acceptable salts thereof provided herein can be used as CDK2 protein inhibitors, effectively inhibiting the activity of CDK2 protein, and thus providing important research value for the preparation of drugs for treating diseases targeting CDK2 protein.
[0083] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the present invention, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the appended claims.
Claims
1. Use of a 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof as a CDK2 protein inhibitor for purposes other than disease treatment, characterized in that: The skeleton of the compound is shown in the following formula (I): in, R 11 H, C 1-10 Alkyl, C 1-10 Alkoxy, i-Pr, halogen, One or more of the following; R 12 H, halogen, C 1-10 One or more alkoxy groups; R 13 H, halogen, C 1-10 One or more alkoxy groups; R 14 H, C 1-10 One or more alkoxy groups; R2 is Ph, C 1-10 One or more of alkoxy-substituted phenyl and halogen-substituted phenyl; R3 is One or more of the .
2. The use according to claim 1, characterized in that R 11 for H, Et, MeO, EtO, i-Pr, F, Cl, One or more of t-Bu, n-BuO; R 12 One or more of H, F, and MeO; R 13 One or more of H, F, Cl, and MeO; R 14 One or more of H, MeO; R2 is Ph, One or more of the following; R3 is One or more of the .
3. Use of a 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and / or treating diseases targeting CDK2 protein, characterized in that: The skeleton of the compound is shown in the following formula (I): in, R 11 H, C 1-10 Alkyl, C 1-10 Alkoxy, i-Pr, halogen, One or more of the following; R 12 H, halogen, C 1-10 One or more alkoxy groups; R 13 H, halogen, C 1-10 One or more alkoxy groups; R 14 H, C 1-10 One or more alkoxy groups; R2 is Ph, C 1-10 One or more of alkoxy-substituted phenyl and halogen-substituted phenyl; R3 is One or more of the .
4. The use according to claim 3, characterized in that R 11 for H, Et, MeO, EtO, i-Pr, F, Cl, One or more of t-Bu, n-BuO; R 12 One or more of H, F, and MeO; R 13 One or more of H, F, Cl, and MeO; R 14 One or more of H, MeO; R2 is Ph, One or more of the following; R3 is One or more of the .
5. The use according to claim 3, characterized in that The 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof is used to inhibit the activity of CDK2 protein.
6. Use of a pharmaceutical composition as a CDK2 protein inhibitor for purposes other than disease treatment, characterized in that: The pharmaceutical composition comprises a 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof represented by the following formula (I), and a pharmaceutically acceptable carrier thereof: in, R 11 H, C 1-10 Alkyl, C 1-10 Alkoxy, i-Pr, halogen, One or more of the following; R 12 H, halogen, C 1-10 One or more alkoxy groups; R 13 H, halogen, C 1-10 One or more alkoxy groups; R 14 H, C 1-10 One or more alkoxy groups; R2 is Ph, C 1-10 One or more of alkoxy-substituted phenyl and halogen-substituted phenyl; R3 is One or more of the .
7. The use according to claim 6, characterized in that R 11 for H, Et, MeO, EtO, i-Pr, F, Cl, One or more of t-Bu, n-BuO; R 12 One or more of H, F, and MeO; R 13 One or more of H, F, Cl, and MeO; R 14 One or more of H, MeO; R2 is Ph, One or more of the following; R3 is One or more of the .
8. Use of a pharmaceutical composition in the preparation of a drug for preventing and / or treating a disease targeting CDK2 protein, characterized in that: The pharmaceutical composition comprises a 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof represented by the following formula (I), and a pharmaceutically acceptable carrier thereof: in, R 11 H, C 1-10 Alkyl, C 1-10 Alkoxy, i-Pr, halogen, One or more of the following; R 12 H, halogen, C 1-10 One or more alkoxy groups; R 13 H, halogen, C 1-10 One or more alkoxy groups; R 14 H, C 1-10 One or more alkoxy groups; R2 is Ph, C 1-10 One or more of alkoxy-substituted phenyl and halogen-substituted phenyl; R3 is One or more of the .
9. The use according to claim 8, characterized in that R 11 for H, Et, MeO, EtO, i-Pr, F, Cl, One or more of t-Bu, n-BuO; R 12 One or more of H, F, and MeO; R 13 One or more of H, F, Cl, and MeO; R 14 One or more of H, MeO; R2 is Ph, One or more of the following; R3 is One or more of the .
10. The use according to claim 8, characterized in that The 4-acetyl-5-phenylpyrrolidine-2,3-dione compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition is used to inhibit the activity of CDK2 protein.