CN113563346A - Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs - Google Patents
Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs Download PDFInfo
- Publication number
- CN113563346A CN113563346A CN202110955578.2A CN202110955578A CN113563346A CN 113563346 A CN113563346 A CN 113563346A CN 202110955578 A CN202110955578 A CN 202110955578A CN 113563346 A CN113563346 A CN 113563346A
- Authority
- CN
- China
- Prior art keywords
- pyrazole
- pyrrole
- usp7
- diamide derivatives
- pyrrolo
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a pyrrole [3,4-c ] pyrazole diamide derivative and application thereof as a USP7 inhibitor in antitumor drugs. The USP7 inhibitor can improve the anti-tumor activity of the drug, has specificity and effectiveness, and has wide development prospect.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a pyrrole [3,4-c ] pyrazole diamide derivative and application thereof as a USP7 inhibitor as an antitumor drug.
Background
Ubiquitin-specific protease 7(USP7) is also called herpes virus-associated ubiquitin-specific protease (HAUSP). It belongs to the ubiquitin-specific protease (USP) family in the deubiquitinase family, and more than 50 USPs are the most abundant family in DUB. Currently deubiquitinating enzymes can be divided into 5 subclasses: ubiquitin-specific proteases (USPs), ubiquitin C-terminal hydrolase (UCH), ovarian tumor proteases (OTU), Machado. Joseph disease proteases (fMJDs), and JABl/MPN/Mov34 metalloproteases (JAMMs). Studies have shown that P53 is an important tumor suppressor, and that P53 is mutated in more than 50% of human cancers. USP7 plays an important role in antagonizing P53 function through multiple mechanisms, wherein MDM2 is one of main E3 ubiquitin ligases, can also be used as a substrate of USP7, the expression level of MDM2 in normal cells is low, MDM2 can be degraded through ubiquitin, and when diseases occur, USP7 in vivo can be over-expressed, so that a USP 7-MDM 2-P53 signal path can be formed, and P53 is down-regulated through the signal path. USP7 is one of the USPs studied more deeply at present, mainly because USP7 has close relation with p53 protein and MDM2 protein, and more than half of cancers are related to abnormal expression of p 53. Besides, USP7 has close connection with tumor-associated proteins and immune system, such as PTEN, FOX04, notch, DNMTl, etc. At present, many diseases are closely related to abnormal expression of USP7, such as ovarian cancer, chronic lymphocytic leukemia, myeloma, breast cancer and the like.
Multiple studies show that USP7 can act on HDM2 protein, thereby influencing the content of p53 in cells and participating in the regulation and control of the apoptosis process; besides, USP7 is also involved in cellular activities such as immunity. For these reasons, inhibitors of USP7 are placing great expectations. Since 2005, the USP7 inhibitor has been widely noticed and developed, but since there is no clear mode of action between active small molecules and USP7, new small molecules can only be screened by conventional methods or functional groups can be explored on the basis of existing methods, the development of USP7 inhibitor is very slow, and after 2017, a large amount of USP7 inhibitor has been developed by means of computer means and eutectic structures. The previous small molecule inhibitors are not high in selectivity and activity, so that the conclusion of some previous biological experiments may not be completely applicable at present, and even the contrary conclusion can be obtained by repeated experiments.
In conclusion, the study of novel, more specific inhibitors of USP7 is of great importance for the clinical treatment of patients with tumor drugs.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a pyrrole [3,4-c ] pyrazole diamide derivative; and a preparation method of the derivative and application of the derivative as a USP7 inhibitor as an antitumor drug.
In order to achieve the purpose, the invention adopts the technical scheme that: the invention provides a pyrrole [3,4-c ] pyrazole diamide derivative shown in a general formula (I),
and n is 1-3.
The R is1、R2Selected from hydrogen, C1-C6Alkyl, alkenyl, alkynyl or aryl; the R is3Selected from hydrogen, halogen, C1-C6Alkyl, alkenyl, alkynyl or aryl.
The 4, 5-dihydronaphthoisoxazole derivative shown in the general formula (I) is selected from the following compounds:
the compound of formula I can be synthesized according to the method of the route 1, substituted 3-amino-5-tert-butoxycarbonyl-pyrrolo [3,4-C ] pyrazole is used as a starting material, and is firstly subjected to condensation reaction with 4-methyl-1-piperazinylpropionic acid/4-methyl-1-piperazineacetic acid in the presence of a condensing agent to obtain an intermediate 2, then subjected to Boc removal under the condition of ethyl acetate hydrochloride to obtain an intermediate 3, and finally subjected to condensation reaction with substituted 3-phenylpropionic acid to obtain a target product.
Synthetic scheme 1 reagents and conditions: (a) HATU, DIEA, DMF, rt.; (b) HCl-EA, rt; (c) HATU, DIEA, DMF, rt.
The pyrrole [3,4-c ] pyrazole diamide derivative can be used as a USP7 inhibitor in application of antitumor drugs.
The tumor cell of the invention can be RS4(ii) a 11 or LNCaP cells.
Detailed Description
The following examples are intended to illustrate but not limit the scope of the invention. The nuclear magnetic resonance hydrogen spectrum of the compound is measured by Bruker ARX-400, and the mass spectrum is measured by Agilent 1100 LC/MS; all reagents used were analytically or chemically pure.
Example 1.
Step 1 Synthesis of intermediate 2
4-methyl-1-piperazinylpropionic acid (0.71g,4.46mmol) was dissolved in DMF, and HATU (2.03g,5.35mmol), DIEA (1.15g,8.92mmol) and 3-amino-5-tert-butoxycarbonyl-pyrrolo [3,4-c ] pyrazole (1.00g,4.46mmol) were added to the solution in this order, and the mixture was heated to 70 ℃ for reaction. After 8 hours, TLC detection reaction was completed, the reaction solution was poured into ice water to precipitate a solid, which was filtered to obtain a crude product, which was then purified by silica gel chromatography to obtain a white-like powdery solid in an amount of 1.06g with a yield of 65.23%.
Step 2 Synthesis of intermediate 3
Dissolving the intermediate 2(1.00g,2.74mmol) in ethyl acetate (10mL), adding a saturated hydrochloric acid-ethyl acetate solution (10mL) under an ice bath condition, gradually heating to room temperature for reaction for 1h, carrying out suction filtration, washing with ethyl acetate (5 mL. times.3), and drying to obtain 0.74g of off-white solid with the yield of 89.66%.
Step 3 Synthesis of example 1
3-phenylpropionic acid (0.25g,1.66mmol) was dissolved in DMF and HATU (0.88g,2.33mmol), DIEA (0.64g,4.99mmol) and intermediate 3(0.50g,1.66mmol) were added to the solution in this order and the reaction was warmed to 70 ℃. After 8 hours, TLC detection reaction was completed, the reaction solution was poured into water to precipitate a solid, which was filtered to obtain a crude product, which was then purified by silica gel chromatography to obtain 0.43g of a white-like powdery solid with a yield of 65.24%.
1H-NMR(400MHz,DMSO-d6)δ12.04(s,1H),10.68(s,1H),7.28–7.23(m,4H),7.19(dd,J＝8.4,2.2Hz,1H),5.12(s,2H),4.46(s,2H),3.11(s,2H),2.88(d,J＝7.0Hz,2H),2.55(d,J＝7.2Hz,2H),2.42(s,3H),2.24–2.12(m,4H).ESI-MS m/z:397.2[M+H]+.
Example 2.
1H-NMR(400MHz,DMSO-d6)δ12.01(s,1H),10.64(s,1H),7.27–7.21(m,4H),7.18(dd,J＝8.2,2.1Hz,1H),5.11(s,2H),4.45(s,2H),3.18–3.11(m,3H),2.58(dd,J＝14.1,9.9Hz,1H),2.42(s,3H),2.31–2.13(m,5H),1.24(d,J＝7.2Hz,3H).ESI-MS m/z:411.2[M+H]+.
Example 3.
1H-NMR(400MHz,DMSO-d6)δ12.03(s,1H),10.60(s,1H),7.26–7.20(m,4H),7.17(dd,J＝8.0,2.2Hz,1H),4.46(s,2H),3.20–3.13(m,3H),2.58(dd,J＝14.4,9.6Hz,1H),2.42(s,3H),2.33–2.23(m,5H),1.44(s,6H),1.24(d,J＝7.0Hz,3H).ESI-MS m/z:439.2[M+H]+.
Example 4.
1H-NMR(400MHz,DMSO-d6)δ12.02(s,1H),10.64(s,1H),7.10(d,J＝8.0Hz,2H),7.01(d,J＝8.4Hz,2H),5.10(s,2H),4.45(s,2H),3.65(t,J＝7.0Hz,2H),2.88(t,J＝7.0Hz,2H),2.56–2.50(m,4H),2.40(s,3H),2.24–2.14(m,4H),2.20(s,3H).ESI-MS m/z:425.2[M+H]+.
Example 5.
1H-NMR(400MHz,DMSO-d6)δ12.02(s,1H),10.64(s,1H),7.28(d,J＝8.4Hz,1H),7.11-7.02(m,3H),5.12(s,2H),4.46(s,2H),3.64(t,J＝7.2Hz,2H),2.87(t,J＝7.0Hz,2H),2.55–2.50(m,4H),2.41(s,3H),2.24–2.15(m,4H),2.24(s,3H).ESI-MS m/z:425.2[M+H]+.
Example 6.
1H-NMR(400MHz,DMSO-d6)δ11.96(s,1H),10.62(s,1H),7.18–7.14(m,4H),5.11(s,2H),4.44(s,2H),3.64(t,J＝7.4Hz,2H),2.86(t,J＝7.1Hz,2H),2.55–2.50(m,4H),2.41(s,3H),2.26–2.15(m,4H).ESI-MS m/z:429.2[M+H]+.
Example 7.
1H-NMR(400MHz,DMSO-d6)δ12.04(s,1H),10.68(s,1H),7.29–7.23(m,4H),7.18(dd,J＝8.0,2.1Hz,1H),5.13(s,2H),4.46(s,2H),3.66(t,J＝7.2Hz,2H),3.18–3.14(m,1H),2.58(dd,J＝14.1,9.9Hz,1H),2.50(t,J＝7.4Hz,2H),2.42(s,3H),2.31–2.24(m,5H),1.24(d,J＝7.1Hz,3H).ESI-MS m/z:425.2[M+H]+.
Example 8.
1H-NMR(400MHz,DMSO-d6)δ12.02(s,1H),10.62(s,1H),7.29–7.24(m,4H),7.16(dd,J＝8.0,2.1Hz,1H),5.11(s,2H),4.47(s,2H),3.66(t,J＝7.4Hz,2H),3.18–3.14(m,1H),2.58(dd,J＝14.0,9.8Hz,1H),2.51(t,J＝7.4Hz,2H),2.42(s,3H),2.33–2.25(m,5H),1.25(d,J＝7.4Hz,3H).ESI-MS m/z:425.2[M+H]+.
Example 9.
1H-NMR(400MHz,DMSO-d6)δ12.01(s,1H),10.65(s,1H),7.27–7.22(m,4H),7.19(dd,J＝8.1,2.2Hz,1H),4.45(s,2H),3.64(t,J＝7.0Hz,2H),3.18–3.14(m,1H),2.56(dd,J＝14.2,9.9Hz,1H),2.50(t,J＝7.4Hz,2H),2.40(s,3H),2.32–2.24(m,5H),1.45(s,6H),1.24(d,J＝7.0Hz,3H).ESI-MS m/z:453.2[M+H]+.
First, pharmacological research
The test method comprises the following steps: the inhibitory activity of the example compounds on USP7 was tested using the Ub-AMC test method. The kit comprises: the USP7 inhibitor screening assay kit divides the experiment into a compound group and a blank combination control group. The buffer was 50mM Tris-HCl (pH 7.6), 0.5mM EDTA, 5mM DTT and 0.05mg/mL BSA. In 384 well plates, 10 μ LUSP7 enzyme (100pM) and compound were incubated at room temperature for 30 minutes, then 10 μ LUb-AMC (300nM) was added, and after further incubation for 2 hours in the dark at room temperature, fluorescence intensity (excitation at 345nM, emission at 445nM) was measured using an Enspire (PerkinElmer) multifunctional microplate reader.
Table 1 the compounds of the examples are active against USP71 inhibition.
| Examples | IC50(μM) | Examples | IC50(μM) |
| Example 1 | 14.51 | Example 6 | 0.64 |
| Example 2 | 20.33 | Example 7 | 0.31 |
| Example 3 | 10.17 | Example 8 | 1.62 |
| Example 4 | 0.97 | Example 9 | 0.44 |
| Example 5 | 4.56 |
Cell proliferation inhibition assay (MTT): tumor cell selection RS4(ii) a 11 and LNCaP cells, culturing the cells to logarithmic growth phase, treating adherent cells by trypsinization, and collecting the cells into DMEM medium containing 10% fetal bovine serum. The cell suspension was centrifuged (1000 Xrpm) and the cells were diluted to 2.5-5.0X 103Adding 2.0-3.0 × 10 per well3The cells were cultured at 37 ℃ for 24 hours. Adding 2 μ L of drug solution with different concentrations, culturing at 37 deg.C, adding 10 μ L of MTT [3- (4, 5-dimethylthiazole-2) -2, 5-diphenyl tetrazolium bromide salt at different time points]And (3) solution. Incubation at 37 ℃ for 4h, medium was discarded, 200 μ L DMSO was added per well to dissolve residual formazan crystals, and after 15min, absorbance was recorded at 490 nm.
Table 1 examples 1-9 inhibitory activity against tumor cells.
Claims (4)
1. Pyrrole [3,4-c ] pyrazole diamide derivatives are characterized in that the structural formula of the derivatives is as follows:
wherein n is 1-3;
the R is1、R2Selected from hydrogen, C1-C6Alkyl, alkenyl, alkynyl or aryl; the R is3Selected from hydrogen, halogen, C1-C6Alkyl, alkenyl, alkynyl or aryl.
2. Pyrrolo [3,4-c ] pyrazole diamide derivatives according to claim 1, selected from:
3. the process for producing a 4, 5-dihydronaphthoisoxazole derivative according to any one of claims 1 to 2, which comprises: taking substituted 3-amino-5-tert-butyloxycarbonyl-pyrrolo [3,4-C ] pyrazole as an initial raw material, firstly carrying out condensation reaction with 4-methyl-1-piperazinylpropionic acid/4-methyl-1-piperazineacetic acid in the presence of a condensing agent to obtain an intermediate 2, then removing Boc under the condition of ethyl acetate hydrochloride to obtain an intermediate 3, and finally carrying out condensation reaction with substituted 3-phenylpropionic acid to obtain a target product.
4. The pyrrolo [3,4-c ] pyrazole diamide derivatives according to any one of claims 1 to 3, which can be used as a USP7 inhibitor in antitumor drugs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110955578.2A CN113563346A (en) | 2021-08-19 | 2021-08-19 | Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110955578.2A CN113563346A (en) | 2021-08-19 | 2021-08-19 | Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113563346A true CN113563346A (en) | 2021-10-29 |
Family
ID=78172152
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110955578.2A Withdrawn CN113563346A (en) | 2021-08-19 | 2021-08-19 | Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113563346A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1447810A (en) * | 2000-08-10 | 2003-10-08 | 法玛西雅意大利公司 | Bicyclo-pyrazoles active as kinase inhibitors,process for their prepn. and pharmaceutical compsns. comprising them |
| CN1820009A (en) * | 2003-07-09 | 2006-08-16 | 法玛西雅意大利公司 | Pyrrolo[3,4-c]pyrazole derivatives active as kinase inhibitors |
| CN101646673A (en) * | 2007-02-07 | 2010-02-10 | 辉瑞大药厂 | 3-amino-pyrrolo[3,4-c] pyrazole- 5 (1h, 4h, 6h) carbaldehyde derivatives as pkc inhibitors |
-
2021
- 2021-08-19 CN CN202110955578.2A patent/CN113563346A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1447810A (en) * | 2000-08-10 | 2003-10-08 | 法玛西雅意大利公司 | Bicyclo-pyrazoles active as kinase inhibitors,process for their prepn. and pharmaceutical compsns. comprising them |
| CN1820009A (en) * | 2003-07-09 | 2006-08-16 | 法玛西雅意大利公司 | Pyrrolo[3,4-c]pyrazole derivatives active as kinase inhibitors |
| CN101646673A (en) * | 2007-02-07 | 2010-02-10 | 辉瑞大药厂 | 3-amino-pyrrolo[3,4-c] pyrazole- 5 (1h, 4h, 6h) carbaldehyde derivatives as pkc inhibitors |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Leonard et al. | Reaction of diethyl pyrocarbonate with nucleic acid components. Adenosine | |
| CN101787025B (en) | Substituted-evodiamine anti-tumor and antifungal compounds and preparation method thereof | |
| CN108117507B (en) | Synthesis method and application of azaspiro cyclohexadienone | |
| WO2022214106A1 (en) | Naphthyl urea compound having anti-cancer effect, preparation method therefor, and use thereof | |
| Schulz et al. | Pyrrolo [1, 2-a] benzimidazole-based quinones and iminoquinones. The role of the 3-substituent on cytotoxicity | |
| CN111559991A (en) | Preparation method and application of naphthylamine compound and salt thereof | |
| KR100588803B1 (en) | Indirubin derivatives having anticancer property against human cancer cell line | |
| CN106146611A (en) | A kind of fluorescent probe substrate measuring dipeptidyl peptidase IV activity and application thereof | |
| CN111747882B (en) | Indole NEDD8 activating enzyme inhibitor, preparation method thereof and application thereof in antitumor drugs | |
| CN102134234A (en) | Indazolyl urea compounds and preparation method and medicinal use thereof | |
| Witiak et al. | Study of trans-cyclopropylbis (diketopiperazine) and chelating agents related to ICRF 159. Cytotoxicity, mutagenicity, and effects on scheduled and unscheduled DNA synthesis | |
| CN112876463B (en) | Intermediate for preparing PD-L1 antagonist and preparation method thereof | |
| CN113563346A (en) | Pyrrole [3,4-c ] pyrazole diamide derivatives and application thereof in antitumor drugs | |
| JP2021508319A (en) | Salt form as an Akt inhibitor and its crystal form | |
| CN113493443B (en) | Application of erlotinib derivative in preparation of medicines for treating esophageal cancer | |
| CN110194741B (en) | 4-benzoyl piperazine-3-nitro-1, 8-naphthalimide derivative and preparation method and application thereof | |
| CN110078706B (en) | Imatinib derivative and preparation method and application thereof | |
| CN110041239B (en) | N- (benzoyl) -L-cysteine methyl ester derivative and preparation method and application thereof | |
| CN108101892B (en) | Chrysin non-natural amino acid derivative and preparation method and application thereof | |
| CN109111376A (en) | A kind of 2,5- double deoxidation strepto- amine derivative and its application | |
| CN116120291B (en) | Indazole compound, preparation method and application thereof | |
| CN112047933B (en) | Quinazolinone USP7 inhibitor and preparation method and application thereof | |
| CN111732590A (en) | 2, 9-disubstituted purine derivative, preparation method thereof and application thereof in antitumor drugs | |
| CN109400597B (en) | VEGFR-2 inhibitor ABT-869-based protein degradation targeting chimera and preparation method and application thereof | |
| CN114478318B (en) | Dinitrile isophorone derivative, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20211029 |
|
| WW01 | Invention patent application withdrawn after publication |