CN101792478A - Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof - Google Patents
Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof Download PDFInfo
- Publication number
- CN101792478A CN101792478A CN201010126952A CN201010126952A CN101792478A CN 101792478 A CN101792478 A CN 101792478A CN 201010126952 A CN201010126952 A CN 201010126952A CN 201010126952 A CN201010126952 A CN 201010126952A CN 101792478 A CN101792478 A CN 101792478A
- Authority
- CN
- China
- Prior art keywords
- compound
- under reduced
- reduced pressure
- solvent under
- ethyl acetate
- 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.)
- Pending
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a light affinity labelling small molecular probe based on maslinic acid and preparation method thereof, belonging to the technical field of organic compound and preparation thereof. Natural product maslinic acid is taken as initial raw material, compound I is obtained under the protection of acetyl, the compound I reacts with oxalyl chloride to obtain compound II, the compound II reacts with ethane diamine under single protection of Boc to obtain compound III, the compound III firstly strips Boc protecting group in HCl ethyl acetate solution and then reacts with segment IV with light affinity labelling group to obtain compound V, the compound V strips acetyl protecting group by hydrolysis with presence of alkali, so as to obtain light affinity labelling small molecular probe VI. The probe molecule can be used for researching and discovering active compound maslinic acid in vivo action target, action mode of target spot and structural information of active site and provides important information for research and development of related medicine.
Description
Technical field
The present invention relates to a kind of photoaffinity labeling small molecules probe based on Crategolic acid and preparation method thereof, belong to the technical field of organise thing and preparation thereof.
Background technology
Drug discovery process comprises many stages, as: the composition optimizes of the discriminating of target spot, the discovery of lead compound, micromolecular compound, the clinical preceding clinical trial etc. that reaches.The discriminating of target spot is the committed step (Curr.Opin.Chem.Biol.2002 in drug discovery and the evolution, 6:427～433), can not to solve which kind of protein be target spot at the small-molecule drug of certain disease to the biotechnology on the gene level and at present.Can predict, the proteomics research of carrying out on the genomics basis will cause the substantive breakthroughs aspect the drug development, make life science can realize its ultimate aim, develop the medicine that treatment comprises the multiple disease of cancer and acquired immune deficiency syndrome (AIDS) etc.Therefore the technology center of gravity at drug discovery has turned to protein group by genome.The photoaffinity labeling technology is one of main policies of research proteomics, and it is not only differentiated protein, also its function and interaction is studied simultaneously.
The photoaffinity labeling technology is in conjunction with multi-disciplinary advantages such as modern molecular biology, cytobiology, organic chemistry, the affine micromolecular compound of utilization synthetic light is as the instrument probe, decompose the highly active intermediate of generation through the rayed of specific wavelength, form specific irreversible covalent attachment with its receptor active position.Probe molecule comprises three functional parts: active group (activity group), light affinity tag group (photoaffinitylabeling group), reporter group (reporter group) or potential reporter group (
Can " click " reaction take place with the vitamin H that is connected with alkynyl or fluorescein such as BODIPY etc. introduce reporter group, ChemBioChem, 2007,8:1351～1358), and connecting portion (spacer).The effect of reactive site is with the lead active centre of receptor protein of probe molecule; The effect of photoaffinity labeling group is to decompose through the UV-irradiation of specific wavelength to produce highly active Cabbeen or nitrene intermediate after probe molecule is directed to the active centre of receptor protein, with the active centre of probe molecule covalent modification at receptor protein; Reporter group or potential reporter group mainly are operations such as convenient later separation, purifying, detection.
Utilization photoaffinity labeling technology is found with target spot disease-related, multiple small molecules is determined with the pattern that biomacromolecule acts in a large number, and these information have very important significance to reasonable medicinal design.Such as the discovery of new drug target, perhaps the function of a known protein is done new elaboration, this meaning for generation, development and the treatment of illustrating disease from molecular level is self-evident.These target spots can be developed into efficiently, the high speed high flux screening model, and a large amount of micromolecular compound of random screening at short notice, find that some more highly active micromolecular compounds are used for further new drug development research as guide's thing.Like this, be starting point from a small molecules probe, its parent compound is perhaps in aspects such as quasi-medicated property, validity even metabolic stability existing problems, but newfound compound then is not subjected to the restriction of original structure, and wideer transformation space is arranged.
Crategolic acid (maslinic acid is a kind of pentacyclic triterpene acid MA), and molecular structure is:
Crategolic acid
maslinic?acid(M)
Mainly be present in the natural phant such as Fructus oleae europaeae, hop, Thinlear Adina Fruit, peppermint, cloves, red date, hawthorn, pomegranate and Salvia japonica Thunb..The MA that studies show that in recent years has many pharmacologically actives, as (life science, 21:264～269) such as anticancer, anti-oxidant, anti-AIDS, antibiotic, anti-inflammatory, anti-diabetes Bs.Crategolic acid has so numerous pharmacologically active, and higher security has caused people's extensive studies interest for human body in addition.
The Wen Xiaoan of China Medicine University etc. has reported Crategolic acid recently, and (glycogenphosphorylase GP) has the inhibition activity (IC of medium tenacity to the glycogen Phosphoric acid esterase
50=28 μ M), they have also tested the hypoglycemic activity of Crategolic acid for diabetes B animal pattern KK-Ay mouse, and the dosage of KK-Ay mouse oral Crategolic acid 10mg/kg every day or 30mg/kg is taken 2 weeks back discovery glucose level continuously and reduced greatly.
Also (Protein tyrosine phosphatase 1B PTP1B) has better inhibited activity (IC to another diabetes target proteins tyrosine-phosphatase 1B and the inventor discovers MA
50=4.26 μ M).Crategolic acid has so numerous physiologically active, in vivo multiple action target spot should be arranged, design, prepare photoaffinity labeling small molecules probe based on Crategolic acid, be expected to be used for research at antidiabetic target spot, be expected in addition to be used to find action target spot in the body of other relevant pharmacologically active such as anticancer, anti-oxidant, anti-AIDS, antibiotic, anti-inflammatory etc. and with the binding mode of relevant target spot, these information all help lend some impetus to the research and development of related drugs.
Summary of the invention
First purpose of the present invention is to propose a kind of photoaffinity probe molecule based on Crategolic acid.This probe molecule can be used for studying, find action target spot in the body of active compound Crategolic acid, and and the binding mode of its target spot, the structural information of avtive spot, for the research and development of related drugs provide important information.This probe molecule is characterised in that and comprises four functional parts: active group, light affinity tag group, potential reporter group and connecting portion, and its molecular structure is:
Tested the inhibition activity (IC of this probe molecule for PTP1B
50=2.30 μ M), find that it suppresses the active inhibition activity (IC that is better than Crategolic acid
50=4.26 μ M).Such probe molecule can be used for studying action target spot in the Crategolic acid body with multiple physiologically active, and the binding mode of target spots various with it, the structural information of avtive spot, for the research and development of related drugs provide important information.
Another object of the present invention is to provide the method for a kind of preparation based on the photoaffinity probe molecule of Crategolic acid.To achieve these goals, the present invention by the following technical solutions.With the natural product Crategolic acid is starting raw material; the ethanoyl protection obtains first Compound I; first Compound I and oxalyl chloride reaction obtain the second Compound I I; the reacting ethylenediamine of the single protection of the second Compound I I and Boc; obtain the 3rd compound III; the 3rd compound III is at first sloughed the Boc protecting group in the ethyl acetate solution of HC1; and then obtain the 5th compound V with the fragment IV reaction that has the photoaffinity labeling group; the 5th compound V hydrolysis in the presence of alkali removes the ethanoyl protecting group, obtains the photoaffinity probe molecule VI of product based on Crategolic acid.
Now describe technical scheme of the present invention in detail.A kind of preparation is characterized in that the concrete operations step based on the method for the photoaffinity labeling small molecules probe of Crategolic acid:
The preparation of the first step first Compound I
Crategolic acid is dissolved in N; dinethylformamide, methylene dichloride or tetrahydrofuran (THF) add acetylation reagent Acetyl Chloride 98Min. or aceticanhydride, add acid binding agent pyridine, triethylamine or 4-Dimethylamino pyridine; room temperature reaction 6 hours; remove solvent under reduced pressure, ethyl acetate, methylene dichloride or chloroform extraction are washed with 10% dilute hydrochloric acid and saturated sodium bicarbonate solution respectively; behind the anhydrous sodium sulfate drying; remove solvent under reduced pressure, through the purification by silica gel column chromatography Compound I of winning, this compound
1H NMR (CDCl
3-d): δ 5.26 (1H, m, H-12), 5.08 (3H, m, H-2), 4.74 (1H, d, J=10.0Hz, H-3), 2.81 (1H, m, H-18), 2.05 (3H, s, CH
3CO), 1.97 (3H, s, CH
3CO), 1.11 (3H, s), 1.04 (3H, s), 0.92 (3H, s), 0.90 (3H, s), 0.89 (6H, s), 0.72 (3H, s);
The preparation of second step, the second Compound I I
First Compound I is dissolved in methylene dichloride, tetrahydrofuran (THF) or chloroform, with acylating reagent oxalyl chloride or thionyl chloride room temperature reaction 12 hours, remove solvent under reduced pressure and get the second Compound I I, do not need purifying, be directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
The second Compound I I that last step reaction obtains is dissolved in methylene dichloride, N; dinethylformamide or tetrahydrofuran (THF); the quadrol that adds the single protection of Boc adds acid binding agent pyridine, triethylamine or 4-Dimethylamino pyridine room temperature reaction 1 hour, removes solvent under reduced pressure after reaction is finished; ethyl acetate, methylene dichloride or chloroform extraction; the saturated common salt washing after drying, removes solvent under reduced pressure; purification by silica gel column chromatography gets the 3rd compound III, this compound
1H NMR (CDCl
3-d): 6.33 (1H, s), 5.37 (1H, s, H-12), 5.10-5.06 (1H, m, H-2), 4.95 (1H, s), 4.74 (1H, d, J=8.1, H-3), 3.41 (2H, m), 3.15 (2H, m), 2.57 (1H, m, H-18), 2.05 (3H, s, CH
3CO), 1.97 (3H, s, CH
3CO), 1.43 (9H, s), 1.20 (3H, s), 1.17 (3H, s), 1.05 (3H, s), 0.90 (3H, s), 0.89 (6H, s), 0.75 (3H, s);
The preparation of the 4th step the 5th compound V
The 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of hydrogenchloride, stirring at room was sloughed the Boc protecting group in 2 hours, remove solvent under reduced pressure, resistates is dissolved in N, dinethylformamide, tetrahydrofuran (THF) or methylene dichloride,, add pyridine or triethylamine, add photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, ethyl acetate, methylene dichloride or chloroform extraction, the saturated common salt washing after drying, removes solvent under reduced pressure and gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V is dissolved in methyl alcohol, add lithium hydroxide or sodium hydroxide, stirring at room 1～8 hour removes solvent under reduced pressure, and dilute hydrochloric acid transfers to PH=6.0 with system, ethyl acetate, methylene dichloride or chloroform extraction, the saturated common salt washing after drying, removes solvent under reduced pressure, purification by silica gel column chromatography must be based on the photoaffinity labeling small molecules probe VI of Crategolic acid, this compound
1H NMR (CDCl
3-d): 8.23 (1H, d, J=8.0), 6.89 (1H, d, J=8.0), 6.80 (1H, s), 6.67 (1H, d, J=8.0), 6.52 (1H, s), 5.36 (1H, s, H-12), 4.11 (2H, t, J=6.2), 3.96 (2H, s), 3.66 (2H, m), 3.54 (2H, m), 3.31 (2H, m), 3.32 (1H, m, H-2), 2.98 (1H, d, J=9.3Hz, H-3), 2.58 (1H, m, H-18), 1.91 (2H, m), 1.69 (2H, m), 1.31 (4H, m), 1.11 (3H, s), 1.02 (3H, s), 0.90 (3H, s), 0.88 (3H, s) 0.85 (3H, s), 0.83 (3H, s), 0.65 (3H, s).
Below be the reaction skeleton symbol relevant with this preparation method:
The invention has the advantages that this probe is expected to be used for research at antidiabetic target spot, be expected in addition to be used to find action target spot in the body of other relevant pharmacologically active such as anticancer, anti-oxidant, anti-AIDS, antibiotic, anti-inflammatory etc. and with the binding mode of relevant target spot, these information all help lend some impetus to the research and development of related drugs.
Embodiment
Now describe technical scheme of the present invention in conjunction with the embodiments in detail.All embodiment operate in strict accordance with above-mentioned preparation method's operation steps.
Embodiment 1
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in 50mL N, dinethylformamide adds the 4.1mL aceticanhydride, adds the 10mL pyridine, room temperature reaction 6 hours, remove solvent under reduced pressure, add 20mL water, dichloromethane extraction 3 times, each 50mL that uses, merge organic phase, wash 2 times, use 20mL at every turn with 10% dilute hydrochloric acid, wash 2 times with saturated sodium bicarbonate solution again and use 20mL at every turn, anhydrous sodium sulfate drying removes solvent under reduced pressure, silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 3.2g first Compound I, yield 91%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 50mL methylene dichloride, adds the 10ml oxalyl chloride, drips 2 N, the dinethylformamide catalyzed reaction, and room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.7g second Compound I I, does not need purifying, is directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in the 20mL methylene dichloride, the quadrol that adds the single protection of 0.6g Boc adds the 0.7mL triethylamine, room temperature reaction 1 hour, remove solvent under reduced pressure, add 20mL water, methylene dichloride divides 3 extractions, uses 60mL at every turn, merge organic phase, 20mL is used in dilute hydrochloric acid washing with 5% 2 times at every turn, saturated sodium bicarbonate solution washing 2 times, each 20mL that uses, anhydrous sodium sulfate drying concentrates silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 1.78g the 3rd compound III, yield 90%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours, remove solvent under reduced pressure, resistates is dissolved in 30mLN, and dinethylformamide adds the 3.2mL triethylamine, add 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, adds 50mL water, ethyl acetate extraction 3 times, each 50mL that uses, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying, remove solvent under reduced pressure, get the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add the 2g sodium hydrate solid, stirring at room 1 hour, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, ethyl acetate extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.7g compound VI, yield 91%.
The first step, the 3rd step, the 5th go on foot
1The contained corresponding data of concrete operations step are identical in HNMR data and the summary of the invention, no longer repeat herein.
Embodiment 2
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in the 50mL methylene dichloride, add the 3.5mL Acetyl Chloride 98Min., add the 10mL triethylamine, room temperature reaction 6 hours removes solvent under reduced pressure, add 20mL water, ethyl acetate extraction 3 times is used 50mL at every turn, merges organic phase, wash 2 times with 10% dilute hydrochloric acid, each 20mL that uses washes 2 times with saturated sodium bicarbonate solution again and uses 20mL, anhydrous sodium sulfate drying at every turn, remove solvent under reduced pressure, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 3.3g first Compound I, yield 94%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 50mL tetrahydrofuran (THF), adds the 10ml thionyl chloride, room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.5g second Compound I I, does not need purifying, is directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in 20mLN, dinethylformamide adds the single quadrol of protecting of 0.6g Boc, add the 0.7mL pyridine, room temperature reaction 1 hour removes solvent under reduced pressure, adds 20mL water, ethyl acetate is divided 3 extractions, each 60mL that uses merges organic phase, the dilute hydrochloric acid washing with 5% 2 times, each 20mL that uses, saturated sodium bicarbonate solution washing 2 times is used 20mL, anhydrous sodium sulfate drying at every turn, concentrate, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 1.8g the 3rd compound III, yield 92%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours, remove solvent under reduced pressure, resistates is dissolved in the 30mL tetrahydrofuran (THF), add the 3.2mL pyridine, add 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours, remove solvent under reduced pressure, add 50mL water, dichloromethane extraction 3 times is used 50mL at every turn, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying removes solvent under reduced pressure, gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add 1.5g lithium hydroxide solid, stirring at room 8 hours, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, dichloromethane extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.8g compound VI, yield 94%.
The first step, the 3rd step, the 5th go on foot
1The contained corresponding data of concrete operations step are identical in H NMR data and the summary of the invention, no longer repeat herein.
Embodiment 3
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in the 50mL tetrahydrofuran (THF), add the 3.5mL Acetyl Chloride 98Min., add the 10mL triethylamine, room temperature reaction 6 hours removes solvent under reduced pressure, add 20mL water, chloroform extraction 3 times is used 30mL at every turn, merges organic phase, wash 2 times with 10% dilute hydrochloric acid, each 20mL that uses washes 2 times with saturated sodium bicarbonate solution again and uses 20mL, anhydrous sodium sulfate drying at every turn, remove solvent under reduced pressure, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 3.0g first Compound I, yield 85%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 40mL chloroform, adds the 10ml thionyl chloride, room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.3g second Compound I I, does not need purifying, is directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in the 20mL tetrahydrofuran (THF), the quadrol that adds the single protection of 0.6g Boc adds the 0.7mL pyridine, room temperature reaction 1 hour, remove solvent under reduced pressure, add 20mL water, chloroform divides 3 extractions, uses 60mL at every turn, merge organic phase, 20mL is used in dilute hydrochloric acid washing with 5% 2 times at every turn, saturated sodium bicarbonate solution washing 2 times, each 20mL that uses, anhydrous sodium sulfate drying concentrates silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 1.7g the 3rd compound III, yield 86%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours removes solvent under reduced pressure, resistates is dissolved in the 30mL methylene dichloride, adds the 3.2mL pyridine, adds 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, adds 50mL water, chloroform extraction 3 times is used 50mL at every turn, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying removes solvent under reduced pressure, gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add 1.5g lithium hydroxide solid, stirring at room 8 hours, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, chloroform extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.6g compound VI, yield 88%.
The first step, the 3rd step, the 5th go on foot
1The contained corresponding data of concrete operations step are identical in HNMR data and the summary of the invention, no longer repeat herein.
Claims (6)
1. the photoaffinity probe molecule based on Crategolic acid is characterized in that, this probe molecule comprises four functional parts: active group, light affinity tag group, potential reporter group and connecting portion, and its molecular structure is:
2. method for preparing based on the photoaffinity probe molecule of Crategolic acid; it is characterized in that; with the natural product Crategolic acid is starting raw material; the ethanoyl protection obtains first Compound I; first Compound I and oxalyl chloride reaction obtain the second Compound I I; the reacting ethylenediamine of the single protection of the second Compound I I and Boc; obtain the 3rd compound III; the 3rd compound III is at first sloughed the Boc protecting group in the ethyl acetate solution of HCl; and then obtain the 5th compound V with the fragment IV reaction that has the photoaffinity labeling group; the 5th compound V hydrolysis in the presence of alkali removes the ethanoyl protecting group, obtains the photoaffinity probe molecule VI of product based on Crategolic acid.
3. preparation according to claim 2 is characterized in that the concrete operations step based on the method for the photoaffinity labeling small molecules probe of Crategolic acid:
The preparation of the first step first Compound I
Crategolic acid is dissolved in N; dinethylformamide, methylene dichloride or tetrahydrofuran (THF) add acetylation reagent Acetyl Chloride 98Min. or aceticanhydride, add acid binding agent pyridine, triethylamine or 4-Dimethylamino pyridine; room temperature reaction 6 hours; remove solvent under reduced pressure, ethyl acetate, methylene dichloride or chloroform extraction are washed with 10% dilute hydrochloric acid and saturated sodium bicarbonate solution respectively; behind the anhydrous sodium sulfate drying; remove solvent under reduced pressure, through the purification by silica gel column chromatography Compound I of winning, this compound
1H NMR (CDCl
3-d): δ 5.26 (1H, m, H-12), 5.08 (3H, m, H-2), 4.74 (1H, d, J=10.0Hz, H-3), 2.81 (1H, m, H-18), 2.05 (3H, s, CH
3CO), 1.97 (3H, s, CH
3CO), 1.11 (3H, s), 1.04 (3H, s), 0.92 (3H, s), 0.90 (3H, s), 0.89 (6H, s), 0.72 (3H, s);
The preparation of second step, the second Compound I I
First Compound I is dissolved in methylene dichloride, tetrahydrofuran (THF) or chloroform, with acylating reagent oxalyl chloride or thionyl chloride room temperature reaction 12 hours, remove solvent under reduced pressure and get the second Compound I I, do not need purifying, be directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
The second Compound I I that last step reaction obtains is dissolved in methylene dichloride, N; dinethylformamide or tetrahydrofuran (THF); the quadrol that adds the single protection of Boc adds acid binding agent pyridine, triethylamine or 4-Dimethylamino pyridine room temperature reaction 1 hour, removes solvent under reduced pressure after reaction is finished; ethyl acetate, methylene dichloride or chloroform extraction; the saturated common salt washing after drying, removes solvent under reduced pressure; purification by silica gel column chromatography gets the 3rd compound III, this compound
1H NMR (CDCl
3-d): 6.33 (1H, s), 5.37 (1H, s, H-12), 5.10-5.06 (1H, m, H-2), 4.95 (1H, s), 4.74 (1H, d, J=8.1, H-3), 3.41 (2H, m), 3.15 (2H, m), 2.57 (1H, m, H-18), 2.05 (3H, s, CH
3CO), 1.97 (3H, s, CH
3CO), 1.43 (9H, s), 1.20 (3H, s), 1.17 (3H, s), 1.05 (3H, s), 0.90 (3H, s), 0.89 (6H, s), 0.75 (3H, s);
The preparation of the 4th step the 5th compound V
The 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of hydrogenchloride, stirring at room was sloughed the Boc protecting group in 2 hours, remove solvent under reduced pressure, resistates is dissolved in N, dinethylformamide, tetrahydrofuran (THF) or methylene dichloride,, add pyridine or triethylamine, add photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, ethyl acetate, methylene dichloride or chloroform extraction, the saturated common salt washing after drying, removes solvent under reduced pressure and gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V is dissolved in methyl alcohol, add lithium hydroxide or sodium hydroxide, stirring at room 1～8 hour removes solvent under reduced pressure, and dilute hydrochloric acid transfers to PH=6.0 with system, ethyl acetate, methylene dichloride or chloroform extraction, the saturated common salt washing after drying, removes solvent under reduced pressure, purification by silica gel column chromatography must be based on the photoaffinity labeling small molecules probe VI of Crategolic acid, this compound
1H NMR (CDCl
3-d): 8.23 (1H, d, J=8.0), 6.89 (1H, d, J=8.0), 6.80 (1H, s), 6.67 (1H, d, J=8.0), 6.52 (1H, s), 5.36 (1H, s, H-12), 4.11 (2H, t, J=6.2), 3.96 (2H, s), 3.66 (2H, m), 3.54 (2H, m), 3.31 (2H, m), 3.32 (1H, m, H-2), 2.98 (1H, d, J=9.3Hz, H-3), 2.58 (1H, m, H-18), 1.91 (2H, m), 1.69 (2H, m), 1.31 (4H, m), 1.11 (3H, s), 1.02 (3H, s), 0.90 (3H, s), 0.88 (3H, s) 0.85 (3H, s), 0.83 (3H, s), 0.65 (3H, s).
4. preparation according to claim 3 is characterized in that the concrete operations step based on the method for the photoaffinity labeling small molecules probe of Crategolic acid:
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in 50mL N, dinethylformamide adds the 4.1mL aceticanhydride, adds the 10mL pyridine, room temperature reaction 6 hours, remove solvent under reduced pressure, add 20mL water, dichloromethane extraction 3 times, each 50mL that uses, merge organic phase, wash 2 times, use 20mL at every turn with 10% dilute hydrochloric acid, wash 2 times with saturated sodium bicarbonate solution again and use 20mL at every turn, anhydrous sodium sulfate drying removes solvent under reduced pressure, silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 3.2g first Compound I, yield 91%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 50mL methylene dichloride, adds the 10ml oxalyl chloride, drips 2 N, the dinethylformamide catalyzed reaction, and room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.7g second Compound I I, does not need purifying, is directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in the 20mL methylene dichloride, the quadrol that adds the single protection of 0.6g Boc adds the 0.7mL triethylamine, room temperature reaction 1 hour, remove solvent under reduced pressure, add 20mL water, methylene dichloride divides 3 extractions, uses 60mL at every turn, merge organic phase, 20mL is used in dilute hydrochloric acid washing with 5% 2 times at every turn, saturated sodium bicarbonate solution washing 2 times, each 20mL that uses, anhydrous sodium sulfate drying concentrates silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 1.78g the 3rd compound III, yield 90%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours, remove solvent under reduced pressure, resistates is dissolved in 30mLN, and dinethylformamide adds the 3.2mL triethylamine, add 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, adds 50mL water, ethyl acetate extraction 3 times, each 50mL that uses, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying, remove solvent under reduced pressure, get the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add the 2g sodium hydrate solid, stirring at room 1 hour, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, ethyl acetate extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.7g compound VI, yield 91%.
5. preparation according to claim 3 is characterized in that the concrete operations step based on the method for the photoaffinity labeling small molecules probe of Crategolic acid:
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in the 50mL methylene dichloride, add the 3.5mL Acetyl Chloride 98Min., add the 10mL triethylamine, room temperature reaction 6 hours removes solvent under reduced pressure, add 20mL water, ethyl acetate extraction 3 times is used 50mL at every turn, merges organic phase, wash 2 times with 10% dilute hydrochloric acid, each 20mL that uses washes 2 times with saturated sodium bicarbonate solution again and uses 20mL, anhydrous sodium sulfate drying at every turn, remove solvent under reduced pressure, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 3.3g first Compound I, yield 94%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 50mL tetrahydrofuran (THF), adds the 10ml thionyl chloride, room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.5g second Compound I I, does not need purifying, is directly used in three-step reaction;
The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in 20mLN, dinethylformamide adds the single quadrol of protecting of 0.6g Boc, add the 0.7mL pyridine, room temperature reaction 1 hour removes solvent under reduced pressure, adds 20mL water, ethyl acetate is divided 3 extractions, each 60mL that uses merges organic phase, the dilute hydrochloric acid washing with 5% 2 times, each 20mL that uses, saturated sodium bicarbonate solution washing 2 times is used 20mL, anhydrous sodium sulfate drying at every turn, concentrate, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 1.8g the 3rd compound III, yield 92%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours, remove solvent under reduced pressure, resistates is dissolved in the 30mL tetrahydrofuran (THF), add the 3.2mL pyridine, add 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours, remove solvent under reduced pressure, add 50mL water, dichloromethane extraction 3 times is used 50mL at every turn, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying removes solvent under reduced pressure, gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add 1.5g lithium hydroxide solid, stirring at room 8 hours, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, dichloromethane extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.8g compound VI, yield 94%.
6. preparation according to claim 3 is characterized in that the concrete operations step based on the method for the photoaffinity labeling small molecules probe of Crategolic acid:
The preparation of the first step first Compound I
3.0g Crategolic acid is dissolved in the 50mL tetrahydrofuran (THF), add the 3.5mL Acetyl Chloride 98Min., add the 10mL triethylamine, room temperature reaction 6 hours removes solvent under reduced pressure, add 20mL water, chloroform extraction 3 times is used 30mL at every turn, merges organic phase, wash 2 times with 10% dilute hydrochloric acid, each 20mL that uses washes 2 times with saturated sodium bicarbonate solution again and uses 20mL, anhydrous sodium sulfate drying at every turn, remove solvent under reduced pressure, silica gel column chromatography, eluent are the mixed solvent of sherwood oil and ethyl acetate, and the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1, get 3.0g first Compound I, yield 85%;
The preparation of second step, the second Compound I I
2.7g first Compound I is dissolved in the 40mL chloroform, adds the 10ml thionyl chloride, room temperature reaction 12 hours removes solvent under reduced pressure and gets the 2.3g second Compound I I, does not need purifying, is directly used in three-step reaction; The preparation of the 3rd step the 3rd compound III
2.0g the second Compound I I is dissolved in the 20mL tetrahydrofuran (THF), the quadrol that adds the single protection of 0.6g Boc adds the 0.7mL pyridine, room temperature reaction 1 hour, remove solvent under reduced pressure, add 20mL water, chloroform divides 3 extractions, uses 60mL at every turn, merge organic phase, 20mL is used in dilute hydrochloric acid washing with 5% 2 times at every turn, saturated sodium bicarbonate solution washing 2 times, each 20mL that uses, anhydrous sodium sulfate drying concentrates silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=3: 1 gets 1.7g the 3rd compound III, yield 86%;
The preparation of the 4th step the 5th compound V
1.2g the 3rd compound III is dissolved in the saturated acetate acetoacetic ester solution of 30mL hydrogenchloride, stirring at room 2 hours removes solvent under reduced pressure, resistates is dissolved in the 30mL methylene dichloride, adds the 3.2mL pyridine, adds 1.5g photoaffinity labeling fragment active ester IV, stirring at room 12 hours removes solvent under reduced pressure, adds 50mL water, chloroform extraction 3 times is used 50mL at every turn, the washing of 50mL saturated common salt, anhydrous sodium sulfate drying removes solvent under reduced pressure, gets the 5th compound V, do not need purifying, be directly used in the reaction of the 5th step;
The 5th step is based on the preparation of the photoaffinity labeling small molecules probe VI of Crategolic acid
The 5th compound V that the 4th step obtained is dissolved in 100mL methyl alcohol, add 1.5g lithium hydroxide solid, stirring at room 8 hours, remove solvent under reduced pressure, with 10% dilute hydrochloric acid system is transferred to PH=6.0, chloroform extraction 3 times is used 50mL at every turn, merges organic phase, the water washing of 50mL saturated common salt, anhydrous sodium sulfate drying concentrates purification by silica gel column chromatography, eluent is the mixed solvent of sherwood oil and ethyl acetate, the mixed solvent ratio is a sherwood oil: ethyl acetate=15: 1 must get the 1.6g compound VI, yield 88%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010126952A CN101792478A (en) | 2010-03-18 | 2010-03-18 | Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010126952A CN101792478A (en) | 2010-03-18 | 2010-03-18 | Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101792478A true CN101792478A (en) | 2010-08-04 |
Family
ID=42585418
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010126952A Pending CN101792478A (en) | 2010-03-18 | 2010-03-18 | Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101792478A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103539764A (en) * | 2012-07-13 | 2014-01-29 | 华南理工大学 | 12-p-methyl benzene acyloxy-14-deoxidized andrographolide photoaffinity labeling molecular probe, and preparation method and pharmaceutical composition thereof |
| CN106085415A (en) * | 2016-07-16 | 2016-11-09 | 北京化工大学 | A kind of berberine affinity labeling probe and preparation method thereof |
| CN109369777A (en) * | 2018-10-18 | 2019-02-22 | 大连理工大学 | A kind of biology affinity probe molecule and the preparation method and application thereof |
| CN109912677A (en) * | 2017-12-12 | 2019-06-21 | 中国科学院大连化学物理研究所 | A kind of ginseng sapoglycoside Rg 3 bioactive molecule probe and synthesis and application based on ABPP |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682740A (en) * | 2005-03-11 | 2005-10-19 | 中国药科大学 | Use of pentacylic triterpene compounds in preparing glycogenic phosphorylase inhibitor |
| CN1908661A (en) * | 2005-08-04 | 2007-02-07 | 中国科学院上海药物研究所 | Structure module of a series small molecule probe design, its preparing process and application |
-
2010
- 2010-03-18 CN CN201010126952A patent/CN101792478A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682740A (en) * | 2005-03-11 | 2005-10-19 | 中国药科大学 | Use of pentacylic triterpene compounds in preparing glycogenic phosphorylase inhibitor |
| CN1908661A (en) * | 2005-08-04 | 2007-02-07 | 中国科学院上海药物研究所 | Structure module of a series small molecule probe design, its preparing process and application |
Non-Patent Citations (1)
| Title |
|---|
| 《生命科学》 20090430 王博等 山楂酸的研究进展 264-269 第21卷, 第2期 2 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103539764A (en) * | 2012-07-13 | 2014-01-29 | 华南理工大学 | 12-p-methyl benzene acyloxy-14-deoxidized andrographolide photoaffinity labeling molecular probe, and preparation method and pharmaceutical composition thereof |
| CN103539764B (en) * | 2012-07-13 | 2015-07-15 | 华南理工大学 | 12-p-methyl benzene acyloxy-14-deoxidized andrographolide photoaffinity labeling molecular probe, and preparation method and pharmaceutical composition thereof |
| CN106085415A (en) * | 2016-07-16 | 2016-11-09 | 北京化工大学 | A kind of berberine affinity labeling probe and preparation method thereof |
| CN106085415B (en) * | 2016-07-16 | 2018-11-16 | 北京化工大学 | A kind of jamaicin affinity labeling probe and preparation method thereof |
| CN109912677A (en) * | 2017-12-12 | 2019-06-21 | 中国科学院大连化学物理研究所 | A kind of ginseng sapoglycoside Rg 3 bioactive molecule probe and synthesis and application based on ABPP |
| CN109369777A (en) * | 2018-10-18 | 2019-02-22 | 大连理工大学 | A kind of biology affinity probe molecule and the preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101307056B (en) | Linear furocoumarin derivates, preparation method and application thereof, pharmaceutical compositions containing the derivates | |
| CN104095859B (en) | The application in preparing anti-hepatic fibrosis medicines of O-(piperidyl) ethyl derivative of Cleistanone Cleistanone | |
| CN103275051B (en) | A kind of 7,3 ', 4 '-trihydroxyflavone derivative and preparing the application in Hepatoma therapy medicine | |
| CN101792438B (en) | Method for synthesizing 1-substituted-1,2,3-tolyltriazole | |
| CN104860993A (en) | Prodrug of flavonoids and application of prodrug | |
| CN104910149A (en) | Palbociclib preparation method | |
| CN101792478A (en) | Light affinity labelling small molecular probe based on maslinic acid and preparation method thereof | |
| CN110028547A (en) | A kind of diosgenin 3-OH derivative and preparation method thereof and medical usage | |
| CN112441952B (en) | Cannabidiol-3-sulfonic acid, preparation method and application thereof, and cannabidiol derivative | |
| CN104447934A (en) | Method for purifying abiraterone acetate | |
| CN104478877A (en) | Ledipasvir intermediate preparation method | |
| CN108341805A (en) | It can be used as the substituted pyridine derivate of C-fms kinase inhibitors | |
| CN103539787B (en) | Isorographolide photoaffinity labeling molecular probe, preparation method and pharmaceutical composition thereof | |
| CN102516219B (en) | Halogenated polyhydroxy xanthene derivatives, preparation method and use thereof | |
| CN102180914A (en) | Preparation method of 2-deoxidizing-D-glucose | |
| CN106866480B (en) | Polycyclic virtue selenide analog derivative and preparation method thereof | |
| CN109336940A (en) | Acridone derivatives and its preparation method and application of the one kind containing galactolipin | |
| Romanucci et al. | Investigation on the solid-phase synthesis of silybin prodrugs and their timed-release | |
| CN102653543A (en) | Preparation method, biological activity and application of 3-substituted type Jolkinolide compound | |
| CN105949139A (en) | Sec-butyl diphenyl tetrazine formamide compound, preparation and application | |
| CN102336746B (en) | Preparation method and anti-cancer action of novel diacid solanesol alkyl tegafur diesters | |
| CN104650160A (en) | Novel synthesis method of capecitabine key intermediate 1,2,3-O-triacetyl-5-deoxy-D-ribose | |
| CN106727571A (en) | The piperazinyl and 1H tetrazole radical derivatives composition of Psiguadial A are used for anti-hepatic fibrosis | |
| CN109400595A (en) | Anticancer compound of the one kind containing thiphene ring | |
| CN103772471B (en) | A kind of estradiol-4-aminomethyl-pyridine conjugate and synthetic method thereof and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100804 |