CN108727338B - 4-piperidone compound and preparation method and application thereof - Google Patents
4-piperidone compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN108727338B CN108727338B CN201810730333.8A CN201810730333A CN108727338B CN 108727338 B CN108727338 B CN 108727338B CN 201810730333 A CN201810730333 A CN 201810730333A CN 108727338 B CN108727338 B CN 108727338B
- Authority
- CN
- China
- Prior art keywords
- disease
- piperidone
- parp1
- cancer
- compound
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing aromatic rings
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Biomedical Technology (AREA)
- Heart & Thoracic Surgery (AREA)
- Urology & Nephrology (AREA)
- Vascular Medicine (AREA)
- Cardiology (AREA)
- Psychology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Physical Education & Sports Medicine (AREA)
- Hospice & Palliative Care (AREA)
- Psychiatry (AREA)
- Ophthalmology & Optometry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Plural Heterocyclic Compounds (AREA)
Abstract
The invention discloses a 4-piperidone compound and a preparation method and application thereof. The compound has the following structural general formula
Description
Technical Field
The invention belongs to the field of preparation of antitumor drugs, and particularly relates to a 4-piperidone compound as well as a preparation method and application thereof.
Background
Dobzhansky in 1946 first proposed the concept of "synthetic lethality", i.e., two genes mutated individually without effect, but at the same time, the mutation would cause cell death. Based on synthetic lethality theory, PARP1 inhibitors are primarily used in patients with BRCA1/2 mutant ovarian or breast cancer. However, with the increase of clinical use, the drug resistance of tumor cells to PARP-1 inhibitors appears, so that the drug effect is reduced and even lost. PARP-1 inhibitor resistance occurs primarily due to: restoring HR function in BRCA1/2 gene mutant tumor cells, resulting in cell survival; secondly, the tumor cells generate multidrug resistance, and the concentration of the PARP-1 inhibitor in the cells is reduced, so that the treatment effect is poor. Olaparib (AZD2281/KU-59436) developed by Aslicon pharmaceutical company is an oral PARP small-molecule inhibitor, shows good development prospect in the research of treating ovarian cancer, breast cancer and solid tumor by combining with medicaments such as cisplatin, carboplatin, paclitaxel and the like, and is on the market at present. However, Olaparib has poor selectivity and inhibitory activity on PARP1, and the effective dose of inhibitory activity at the cellular level is 200. mu.M, and the in vivo dose is more than 100mg, which shows significant antitumor activity. The clinical daily dose is up to 400mg (50mg capsule, 8 capsules). Short duration of action and half-life in vivo (<1 hour) and low bioavailability (< 15%). Therefore, there is a need to synthesize more efficient novel PARP1 inhibitors to kill tumor cells.
Heat shock 90(HSP90) is a known target for anti-tumor effect, and has more than 200 client proteins closely related to tumor cell proliferation and metastasis, wherein BRCA1 is also a client protein of HSP 90. The inhibition of HSP90 can promote the degradation of BRCA1 by proteasome, enhance the anti-tumor activity of PARP-1 inhibitor and overcome the drug resistance of PARP-1 inhibitor. Based on the fact, the HSP90/PARP-1 double-target-point drug can overcome the drug resistance of the PARP-1 inhibitor, expand the applicable population of the PARP-1 inhibitor and has important significance for clinical treatment of solid tumors such as ovarian cancer, breast cancer and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a 4-piperidone compound and a preparation method and application thereof, and the invention designs and synthesizes the 4-piperidone by using a molecular docking virtual screening technology and according to a split principle, an electronic isostere principle and ring system transformation on the basis of summarizing the structure-activity relationship between an PARP1 inhibitor Olaparib (AZD2281) and a 3, 5-diarylmethyl-4-piperidone Hsp90 inhibitor, wherein the compound has stronger Hsp90 and PARP1 double-target point inhibitory activity and has good curative effect on treating cancers, neurodegenerative diseases or ischemic diseases.
A 4-piperidone compound with the following structural general formula
Wherein Ar is a benzene ring or an aromatic heterocycle.
The improvement is that the substituent on the benzene ring is hydroxyl, halogen or alkoxy; the aromatic heterocyclic ring is pyridine, quinoline or pyrimidine.
The preparation method of the 4-piperidone compound comprises the steps of condensing 2-2-fluoro-5- [ (4-oxo-3, 4-dihydronaphthyridin-1-yl) methyl ] benzoic acid serving as an intermediate of olaparic acid and 4-piperidone under the action of a condensing agent to obtain an intermediate, reacting the intermediate with aromatic aldehyde under an alkaline condition, extracting a reaction product by using dichloromethane, washing by using water, concentrating, and passing through a silica gel column to obtain the 4-piperidone compound; the volume ratio of ethyl acetate to petroleum ether used for the silica gel column is 3: 1.
the reaction formula is as follows:
as an improvement, the condensing agent is prepared by mixing O-benzotriazole-tetramethyluronium Hexafluorophosphate (HBTU), 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylethylamine (DIPEA) according to a molar ratio of 1.3: 0.3: 3, and mixing.
A pharmaceutical composition contains 4-piperidone compounds or their acceptable salts as active ingredient.
The application of the pharmaceutical composition in preparing products for preventing or treating diseases related to PARP 1.
A PARP1 inhibitor contains 4-piperidone compounds or their acceptable salts as active ingredient.
The PARP1 inhibitor is applied to the preparation of products for preventing or treating diseases related to PARP 1.
As an improvement, the PARP 1-related diseases include cancer, neurodegenerative diseases or ischemic diseases.
In a further refinement, the cancer is breast cancer, ovarian cancer, liver cancer, melanoma, prostate cancer, colon cancer, or stomach cancer; the neurodegenerative disease comprises Parkinson's disease, Alzheimer's disease or muscular dystrophy; the ischemic disease includes cerebral, umbilical cord, heart, digestive tract or retinal ischemic disease.
Has the advantages that:
the compound of the invention has strong PARP1 and HSP90 inhibitory activity, and can be used for preventing and treating diseases caused by PARP1 abnormality and HSP90 high expression or high activity, including cancer, neurodegenerative diseases or ischemic diseases; the simultaneous inhibition of the two targets can enhance the drug effect, reduce the administration dosage, reduce the toxic and side effects and effectively overcome the problem that the single-target inhibitor is easy to generate drug resistance.
Detailed Description
The present invention will be described in further detail below with reference to specific examples.
Example 1
Preparation of N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
The olaparinic acid intermediate (50mg, M297.6, 0.168mmol), HBTU (82mg, M379.24, 0.2184mmol), HOBT (6.8mg, M135.12, 0.05mmol) were dissolved in 5ml dry THF (plus 2ml dmf co-solvent), stirred at room temperature for half an hour, DIPEA (0.86ml,65.14mg,0.504mmol, M129.24,
) Then, 22.32mg of 4-piperidone (M: 148.05 hydrochloric acid form 0.1508mmol) was added thereto, and the mixture was stirred at 50 ℃ overnight. Extracting with dichloromethane, washing with water for 3 times, and washing with brine to obtain intermediate 1, i.e. N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl]-2-fluorobenzoyl]Piperidin-4-one.
Ethyl acetate: the petroleum ether-3: 1 column chromatography gave 25.79 mg. The yield was 40%. And Mp: 230.8-233.8 ℃ C. 1HNMR (400MHz, Chloroform-d)10.94(s,1H),8.51(s,1H),7.81-7.76(M,3H),7.39(d,2H),7.09(M,1H),4.33(s,2H),4.20-3.92(M,2H),3.63(s,2H),2.61(M,2H),2.46(s,2H). LC-MS (ESI) M/z 380.1[ M ESI ]+H]+。
Example 2
Preparation of 3,5- (E) -bis (2-pyridylmethenyl) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
After 155.28 mg (0.146mmol, M ═ 379.13), 1.67mg (0.02mmol, M ═ 84.01) of sodium hydroxide and 10ml of absolute ethanol were mixed, the mixture was stirred at room temperature, and after clarification, 44.6mg (0.4166mmol, M ═ 107.11) of 2-pyridinecarboxaldehyde was added and the reaction was carried out at room temperature. Dichloromethane extraction, water washing 3 times, brine washing once. To obtain the 3,5- (E) -bis (2-pyridylmethenyl) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl group]-2-fluorobenzoyl]Piperidin-4-one. Ethyl acetate: the petroleum ether-3: 1 column chromatography gave 20.79 mg. Yield: 37 percent. And Mp: 156.3-158.0 ℃.1H NMR (400MHz, Chloroform-d)10.99(s,1H),8.79(s,1H),8.46(s,1H),8.18(s,1H),7.63(M,9H),7.38(d,1H),7.10(d,3H),6.81(s,1H),5.52(s,2H),5.24(s,2H),4.08(s,2H), LC-MS (ESI) M/z 558.2[ M+H]+。
Example 3
Preparation of 3,5- (E) -bis (3-pyridylmethenyl) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and 3-pyridinecarboxaldehyde 44.6mg (0.4166mmol, M ═ 107.11) by the same method as in experimental example 2, giving 29.6mg of a yellow solid. The yield was 52%. And Mp: 147.39-149.7 ℃.1H NMR (400MHz, Chloroform-d)11.52(s,1H),8.78(s,1H),8.66(d,1H), 8.62-8.52 (m,1H),8.49(s,1H),8.41(s,1H),7.88(s,2H),7.80(s,3H),7.72(s,2H),7.54(s,1H),7.44(s,1H),7.17(d,2H),6.80(s,1H),5.09(s,2H),4.63(s,2H),4.17(s,2H).LC-MS(ESI)m/z:556.21[M+H]+。
Example 4
Preparation of 3,5- (E) -bis (4-pyridylmethenyl) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and 4-pyridinecarboxaldehyde 44.6mg (0.4166mmol, M ═ 107.11) by the same method as in experimental example 2, giving 20.6mg of a yellow solid. Mp 202.4 ℃ -204.3 ℃ ESI-MS, M/z [ M-H ]: 556.11.1H NMR (400MHz, Chloroform-d)10.58(s,1H), 8.84-8.69 (m,1H),8.62(s,1H),8.47(dd, J ═ 21.8,12.8Hz,2H),8.04(s,3H), 7.91-7.60 (m,4H),7.37(d, J ═ 16.8Hz,1H),7.13(s,2H),6.99(d, J ═ 41.6Hz,2H),6.81(s,1H),5.06(s,1H),4.54(s,1H),4.16(s,2H), 3.61-3.25 (m, 2H).
Example 5
Preparation of 3,5- (E) -bis (2-fluorobenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and 51.67mg (0.4166mmol, M ═ 124.01) of 2-fluorobenzaldehyde, and was prepared as in experimental example 2 to give 29.6mg of a yellow solid. Mp 130.2-130.8 ℃.1H NMR (400MHz, Chloroform-d)10.72(s,1H), 8.56-8.38 (M,1H),8.03(d,1H),7.83(s,1H), 7.81-7.72 (M,2H), 7.69-7.57 (M,1H), 7.55-7.34 (M,2H),7.26(d,2H), 7.23-7.12 (M,2H), 7.13-6.81 (M,4H),6.72(t,1H),4.98(s,2H),4.43(s,2H),4.12(s,2H), LC-MS (ESI) M/z 590.19[ M,1H ], [ M+H]+。
Example 6
Preparation of 3,5- (E) -bis (3-fluorobenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and 51.67mg (0.4166mmol, M ═ 124.01) of 3-fluorobenzaldehyde, and prepared as in experimental example 2 to give 29.6mg of a yellow solid. Mp is 157.8-162.7 deg.C. 1H NMR (400MHz, Chloroform-d)10.83(s,1H), 8.75-8.27 (m,1H),7.85(s,1H), 7.80-7.69 (m,3H), 7.69-7.57 (m,1H), 7.52-7.37 (m,1H),7.31(d,1H), 7.27-7.06 (m,5H), 7.08-6.93(m,1H),6.88(d,1H),6.86–6.65(m,2H),5.09(s,2H),4.56(s,2H),4.14(s,2H).LC-MS(ESI)m/z:590.16[M+H]+.Mp:157.8-162.7℃。
Example 7
Preparation of 3,5- (E) -bis (4-fluorobenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and p-fluorobenzaldehyde 51.67mg (0.4166mmol, M ═ 124.03) and was prepared as in experimental example 2 to give 20.6mg of a yellow solid. Mp is 226.4-231.3 ℃.1H NMR (400MHz, Chloroform-d)10.28(s,1H), 8.66-8.27 (M,1H),7.85(s,2H), 7.77-7.71 (M,3H),7.65(d, J ═ 5.9Hz,2H),7.23(d, J ═ 15.3Hz,2H),6.87(d, J ═ 17.5Hz,1H),6.81(s,2H),6.45(s,2H),4.72(s,2H),4.11(s,2H),3.88(s,2H), LC-ms (esi) M/z 590.11[ M,2H ], [ M+H]+。
Example 8
Preparation of 3,5- (E) -bis (4-chlorobenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and p-chlorobenzaldehyde 51.67mg (0.4166mmol, M ═ 140) by the same method as in experimental example 2, giving 20.6mg of a yellow solid. Mp:223.4-226.3 ℃.1H NMR (400MHz, Chloroform-d)10.50(s,1H), 8.60-8.39 (M,1H),7.84(s,1H),7.76(M,3H),7.65(d, J ═ 4.9Hz,1H),7.46(d, J ═ 9.3Hz,4H),7.20(d, J ═ 5.9Hz,4H),7.01(d, J ═ 7.7Hz,2H),6.80(t, J ═ 8.6Hz,1H),5.05(s,2H),4.55(s,2H),4.13(s,2H), LC-ms (esi) M/z 624.17[ M, J ═ 8.6Hz,1H ], 5.05(s,2H),4.55(s,2H),4.13(s,2H) ]+H]+。
Example 9
Preparation of 3,5- (E) -bis (3, 4-dioxomethyl) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and piperonal 51.67mg (0.4166mmol, M ═ 150.03) by the same method as in experimental example 2, giving 20.6mg of a yellow solid. Mp is 109.4-110.0 deg.C. 1H NMR (400MHz, Chloroform-d)10.37(s,1H),8.49(d, J ═ 7.1Hz,2H),8.04(s,1H), 7.83-7.75 (m,4H), 7.43-7.37 (m,2H), 7.17-7.01 (m,2H), 7.01-6.88 (m,2H),6.70(d,J＝5.8Hz,1H),6.54(s,1H),6.06(s,2H),6.05–5.88(m,2H),4.57(s,2H),4.41–4.19(m,4H).LC-MS(ESI)m/z:644.17[M+H]+。
example 10
Preparation of 3,5- (E) -bis (3,4, 5-trimethoxybenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13), and 3,4, 5-trimethoxybenzaldehyde 44.6mg (0.4166mmol, M ═ 196.07), and prepared as in experimental example 2, to give 22.6mg of a yellow solid. And Mp:>250℃。1H NMR(400MHz,Chloroform-d)10.28(s,1H),8.66–8.27(m,1H),7.85(s,1H),7.77–7.71(m,3H),7.65(d,1H),7.23(d,2H),6.87(d,1H),6.81(s,2H),6.45(s,2H),4.72(s,2H),4.11(s,2H),3.94(d,9H),3.91(s,4H),3.88(s,2H),3.79(s,5H).LC-MS(ESI)m/z:734.24[M-H]+。
example 11
Preparation of 3,5- (E) -bis (2-hydroxybenzylidene) -N- [5- [ (3, 4-dihydro-4-oxo-1-phthalazinyl) methyl ] -2-fluorobenzoyl ] piperidin-4-one
This product was prepared from intermediate 155.28 mg (0.146mmol, M ═ 379.13) and salicylaldehyde 51.67mg (0.4166mmol, M ═ 150.03) by reaction with 10ml of saturated HCl in glacial acetic acid at room temperature overnight. Dichloromethane extraction, water washing 3 times, brine washing once. Ethyl acetate: the white solid 20.6mg is obtained by 3:1 column chromatography. Mp is 202.4-204.3 ℃.1HNMR (400MHz, Chloroform-d)10.78(s,1H),8.51(d,1H),7.78(d,3H),7.40(M,2H),7.33(d,1H),7.23(s,3H),7.09(M,3H),7.01(s,1H),6.85(M,2H),6.70(s,1H),4.84(d,1H),4.33(s,2H),4.26(d,2H),4.04(d,1H),3.51(d,2H), LC-MS (ESI) M/z 586.19[ M, M+H]+。
Example 12
Cell lines: SKBR3 (from Shanghai cell bank), MDA-MB-231 (from Shanghai cell bank), MX-1(BRCA1-/-)(from the institute of biotechnology, Chuanglian, Beijing).
The experimental method comprises the following steps: MTT method. Succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT into water-insoluble blue-violet crystalline Formazan (Formazan) and deposit the Formazan in the cells, but dead cells do not have the function. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and the light absorption value of the formazan is measured at the wavelength of 570nm by using an enzyme-labeling instrument. Within a certain range of cell number, MTT crystals are formed in an amount proportional to the cell number. The number of living cells is judged according to the measured absorbance value (OD value), and the smaller the OD value, the weaker the cell activity, the higher the drug toxicity.
The specific method comprises the following steps: 4000 cells/hole of MCF-7 cells, 8000 cells/hole of SKBR3 cells, 10000 cells/hole of MDA-MB-231 cells and MX-1 cells of tumor cells in logarithmic growth phase are taken(BRCA1-/-)2000 cells/well, HCC1937(BRCA1-/-)Cells were seeded at a density of 8500 cells/well in 96-well culture plates at 180. mu.L per well. After overnight incubation, drug at different concentrations (final concentration starting at 100uM, 6-fold dilution gradient) was added for 48h, three duplicate wells were set for each concentration, and a dimethyl sulfoxide (DMSO) vehicle control and a cell-free zeroing well were set at the corresponding concentrations. After the action is finished, target compounds with different concentrations are respectively added into experimental groups, no medicine is added into a control group, each group is provided with 3 parallel holes, and the culture is carried out for 48 hours at 37 ℃. 5mg/mL MTT solution 20. mu.L/well was added, incubation was continued at 37 ℃ for 4 hours, the supernatant was discarded, 150. mu.L DMSO was added, formazan was dissolved by shaking, and OD570 was measured by a microplate reader.
The degree of inhibition of cell proliferation by the drug was calculated according to the following formula:
inhibition ratio (%) ═ (OD)Control-ODMedicine feeding hole)/ODControl well×100％
And calculating the concentration of the drug at 50% inhibition, i.e., IC, according to the Logit method50The value is obtained. The experiment was repeated three times and the average was calculated.
TABLE 14 IC of inhibitory Effect of piperidones on tumor cell proliferation50Value of
The in vitro cell proliferation inhibition experiment shows that the compound of the invention is used for human breast cancer cell strain MX-1(BRCA1-/-)、HCC1937(BRCA1-/-)The compound has good inhibitory activity, but has weak inhibitory activity on BRCA-free human breast cancer cell lines MCF-7, SKBR3 and MDA-MB-231, which shows that the designed compound has high selectivity on BRCA-deficient cells, wherein the compounds of example 2, example 3, example 4, example 6 and example 8 have strong inhibitory action on various breast cancer cell lines, and the compounds also have other effects different from PARP1 inhibitors.
Example 13
Detection Using cell-free PARP1 enzymatic reaction System
A cell-free PARP1 enzymatic reaction system is constructed by utilizing a PARP1 activity detection kit, and the reaction system can activate the activity of PARP1 enzyme in an in vitro environment.
The PARP1 activity assay kit used in the examples was purchased from Trevigen, USA, model 4677-096-K.
The PARP1 activity detection kit comprises buffer solution, PARP1 recombinant protein and NAD+Single-stranded fragmented DNA, recombinant protein 1(Histone H1).
A cell-free PARP1 enzymatic reaction system is configured according to a PARP1 activity detection kit, and the specific operation steps are as follows: 50ng of PARP1 recombinant protein (PARP protein) was added to the buffer to a final concentration of 10mmol/L NAD+And the final concentration was 20mg/ml single-stranded fragmented DNA. After preparation, co-incubation was performed at room temperature for 60 min.
In the above cell-free PARP1 enzymatic reaction system, recombinant Histone 1(Histone H1) was selected as a target protein (Histone H1 is a target protein of PARP1 recombinant protein, and can be modified by poly ADP ribosylation), and 3-aminobenzamide (3-AB) and Olaparib (Olaparib) were selected as PARP1 inhibitors and simultaneously as positive control drugs.
Adding NAD into the reaction system+First, 3-aminobenzamide (3-AB) was added to the cells to a final concentration of 10mmol/L for preincubation for 5 minutes, and then poly ADP ribosylation was performed according to the above cell-free PARP1 enzymatic reaction system. Examination ofThe test drug groups were added with the compounds prepared in examples 1 to 11 at different concentration gradients, respectively, to conduct poly ADP ribosylation. After the incubation of the system was completed, PARP1 enzyme activity assay was started.
The PARP1 activity is determined by using a PARP1 activity detection kit, and the specific detection steps are shown in the specification of the PARP1 activity detection kit. Among them, 3-AB and Olaparib (10mmol/L) were used as positive controls.
TABLE 24 inhibition of PARP1 enzymatic Activity at the molecular level by piperidones
| EXAMPLES Compounds | Molecular level PARP1(nM) |
| 1 | <50 |
| 2 | <10 |
| 3 | <10 |
| 4 | <10 |
| 5 | <20 |
| 6 | <20 |
| 7 | <50 |
| 8 | <20 |
| 9 | <20 |
| 10 | <30 |
| 11 | <50 |
| Olaparib | <50 |
| 3-AB | <50000 |
From the table, we can see that the 4-piperidones compound has high affinity to PARP1 enzyme at molecular level, has obvious inhibitory activity to PARP1, has nanomolar concentration (< 50nM) of the compound inhibition rate, and has stronger inhibitory activity to PARP1 than the positive compounds Ola and 3-AB. Therefore, the compound of the present invention can be used as a potent and novel PARP-1 inhibitor for preventing and treating PARP (ribose poly ADP-ribose polymerase) related diseases, such as ischemic diseases, neurodegenerative diseases and cancers.
BIACORE method for detecting affinity of 4-piperidone compound and HSP90 protein
1. Coupling the results with the exploration of pH conditions
HSP90 protein was coupled to CM5 chips using the Immobilization pH Scouting protocol in Biacore T200 control software Find Immobilization pH. HBS-EP as working buffer HSP90 protein was diluted to a final concentration of about 30. mu.g/mL with 10nM sodium acetate buffer at pH 5.5, 5.0, 4.5, 4.0, respectively. The results of multiple coupling experiments show that the sodium acetate buffer solution with the pH value of 4.0 can completely meet the coupling requirement and is suitable for coupling HSP90 protein.
2. Coupling of
HSP90 was separately coupled to CM5 chips using amino coupling in the Biacore T200 control software Kinetics and Affinity. HBS-EP was used as a working buffer and HSP90 protein was diluted to a final concentration of about 30. mu.g/mL with 10nM/L sodium acetate buffer at pH 4.0. The chip surface was mixed with 0.4M 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide aqueous solution and 0.1. 0.1M N-hydroxysuccinimide aqueous solution at a ratio of 1: 1, injected at a flow rate of 10. mu.L/min for 7min, then injected with HSP90 protein solution, injected with 1M ethanolamine-HCl blocking solution for 7min, and the activated chip surface was blocked.
3. Kinetic determination
10mM 4-piperidones, STA9090 and Olaparib were diluted to different concentrations (3.125, 6.25, 12.5, 25, 50, 100, 200. mu.M) with Running buffer containing 2% DMSO, respectively. Glycine-HCl 2.0 solution was used as regeneration liquid. Kinetic experiments were performed with a kinetic analysis Wizard in Biacore T200 control software, respectively. Fitting is carried out according to Biacore T200 analysis software to obtain the KD value of the sample, and the smaller the KD value, the stronger the affinity of the sample and the HSP90 protein is. For the real-time sensorgrams obtained in the experiments, the action patterns of Langmuir with the interaction molecule ratio of 1: 1 were selected for fitting by using the Kinetics/Affinity analysis in the software BIAevaluation, and the KD values of the 4-piperidone drugs, STA9090 and Olaparib are shown in Table 3 below.
TABLE 34 affinity of piperidones for Hsp90 protein
Hsp90 protein is coupled on a CM5 chip, the binding condition of the piperidone and Hsp90 protein is detected, and the result shows that 4-piperidone and H are combined by taking an Hsp90 protein inhibitor STA9090 as a positive control drugsp90 protein can be combined, wherein the combination of example 3, example 4, example 10, example 11 and Hsp90 protein is stronger, and the concentration is as low as 10-5M, while Olaparib hardly combined with Hsp90 protein, indicating that 4-piperidone has PARP1 and Hsp90 dual-target effect, which is superior to Olaparib.
The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, and any simple modifications or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.
Claims (9)
1. The 4-piperidone compound is characterized by having the following structural general formula
Wherein Ar is
2. The preparation method of 4-piperidone compounds as claimed in claim 1, characterized in that the olaparinic acid intermediate 2-fluoro-5- [ (4-oxo-3, 4-dihydrophenazine-1-yl) methyl ] benzoic acid and 4-piperidone are condensed under the action of a condensing agent to obtain an intermediate, then the intermediate is reacted with aromatic aldehyde under alkaline conditions, and the reaction product is extracted with dichloromethane, washed with water, concentrated and passed through a silica gel column to obtain 4-piperidone compounds; the volume ratio of ethyl acetate to petroleum ether used by the silica gel column is 3: 1.
3. the method for preparing 4-piperidones as claimed in claim 2, wherein the condensing agent is prepared from O-benzotriazole-tetramethylurea hexafluorophosphate, 1-hydroxybenzotriazole and N, N-diisopropylethylamine in a molar ratio of 1.3: 0.3: 3, and mixing.
4. A pharmaceutical composition, the active ingredient is the 4-piperidone compound as described in claim 1.
5. The use of the pharmaceutical composition of claim 4 in the preparation of a product for preventing or treating a disease associated with PARP 1.
6. A PARP1 inhibitor, wherein the active ingredient is the 4-piperidone compound as defined in claim 1.
7. Use of the PARP1 inhibitor of claim 6 for the preparation of a product for the prevention or treatment of PARP1 related diseases.
8. The use of claim 5 or 7, wherein the PARP1 related disease is cancer, neurodegenerative disease or ischemic disease.
9. The use of claim 8, wherein the cancer is breast cancer, ovarian cancer, liver cancer, melanoma, prostate cancer, colon cancer, or stomach cancer; the neurodegenerative disease is Parkinson's disease, Alzheimer's disease or muscular dystrophy; the ischemic disease is cerebral ischemic disease, umbilical cord ischemic disease, heart ischemic disease, digestive tract ischemic disease or retina ischemic disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810730333.8A CN108727338B (en) | 2018-07-05 | 2018-07-05 | 4-piperidone compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810730333.8A CN108727338B (en) | 2018-07-05 | 2018-07-05 | 4-piperidone compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108727338A CN108727338A (en) | 2018-11-02 |
| CN108727338B true CN108727338B (en) | 2020-12-15 |
Family
ID=63926360
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810730333.8A Active CN108727338B (en) | 2018-07-05 | 2018-07-05 | 4-piperidone compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108727338B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788000A (en) * | 2003-03-12 | 2006-06-14 | 库多斯药物有限公司 | Phthalazinone derivatives |
| CN101925595A (en) * | 2008-01-23 | 2010-12-22 | 阿斯利康(瑞典)有限公司 | Phthalazinone derivatives |
| WO2014182744A1 (en) * | 2013-05-08 | 2014-11-13 | The Johns Hopkins University | Novel bis-benzylidine piperidone proteasome inhibitor with anticancer activity |
-
2018
- 2018-07-05 CN CN201810730333.8A patent/CN108727338B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788000A (en) * | 2003-03-12 | 2006-06-14 | 库多斯药物有限公司 | Phthalazinone derivatives |
| CN101925595A (en) * | 2008-01-23 | 2010-12-22 | 阿斯利康(瑞典)有限公司 | Phthalazinone derivatives |
| WO2014182744A1 (en) * | 2013-05-08 | 2014-11-13 | The Johns Hopkins University | Novel bis-benzylidine piperidone proteasome inhibitor with anticancer activity |
Non-Patent Citations (2)
| Title |
|---|
| Novel synthetic curcumin analogues EF31 and UBS109 are potent DNA hypomethylating agents in pancreatic cancer;Ganji Purnachandra Nagaraju et al.;《Cancer Letters》;20131231;第341卷;195-203 * |
| Synthesis of novel dual target inhibitors of PARP and HSP90 and their antitumor activities;Shanshan Lin et al.;《Bioorganic & Medicinal Chemistry》;20200313;第28卷;115434 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108727338A (en) | 2018-11-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104080455B (en) | Certain chemical entities, composition and method | |
| US10793543B2 (en) | Selective C-KIT kinase inhibitor | |
| CN107245056A (en) | Chemical entities, composition and method | |
| CN107750251A (en) | The imidazopyridine of isoxazolyl substitution | |
| CN105189456A (en) | Covalent inhibitors of KRAS G12C | |
| CN104812389A (en) | Certain chemical entities, compositions, and methods | |
| TW201245161A (en) | 1-(arylmethyl)quinazoline-2,4(1H,3H)-diones as PARP inhibitors and the use thereof | |
| CN105524058A (en) | Pyrazolo[1, 5-a]pyridine compound and use thereof | |
| CN103420917B (en) | Containing the benzamide compound of condensed cyclic structure and apply as antitumor drug | |
| CN108530310A (en) | 2- (the miscellaneous base of substituted benzene) fragrance formic acid class FTO inhibitor, preparation method and its application | |
| CN105461699A (en) | Substituted heterocyclic compound, and use method and use thereof | |
| CN106456580A (en) | Heterocyclic hydroxamic acids as protein deacetylase inhibitors and dual protein deacetylase-protein kinase inhibitors and methods of use thereof | |
| CN107698603A (en) | Thienopyrimidines, its preparation method, Pharmaceutical composition and its application | |
| CN107660202A (en) | water-soluble prodrug | |
| CN107245075A (en) | Simultaneously [3,4 d] pyrimidines and its salt and the application of 2,4,6 3 substituted pyridines | |
| WO2022199547A1 (en) | 7,9-dihydropurine derivative and pharmaceutical purpose thereof | |
| WO2013032797A2 (en) | Oxetane 3,3-dicarboxamide compounds and methods of making and using same | |
| CN103420923B (en) | 4-amido quinazoline hydroxamic acid compound and applying as antitumor drug | |
| CN103992236A (en) | New targeting antitumor drug and preparation method and application thereof | |
| CN108727338B (en) | 4-piperidone compound and preparation method and application thereof | |
| CN114773356B (en) | Sesquiterpene derivative, pharmaceutical composition thereof, and preparation method and application thereof | |
| CN112912075A (en) | Combination therapy for the treatment of prostate cancer | |
| CN108314676B (en) | Amino pyridine derivative containing hydroxamic acid fragment and anti-tumor application thereof | |
| CN112979659B (en) | Preparation and application of HIF-2 alpha small molecule inhibitor | |
| CN102617478B (en) | Synthesis of benzimidazole, oxazole and thiazole derivatives 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |