CN106366075B - Pyrrolidines, salt, nuclear-magnetism probe and medicinal application, reagent, drug - Google Patents
Pyrrolidines, salt, nuclear-magnetism probe and medicinal application, reagent, drug Download PDFInfo
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- CN106366075B CN106366075B CN201610695445.5A CN201610695445A CN106366075B CN 106366075 B CN106366075 B CN 106366075B CN 201610695445 A CN201610695445 A CN 201610695445A CN 106366075 B CN106366075 B CN 106366075B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4192—1,2,3-Triazoles
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
- A61K49/101—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals
- A61K49/106—Organic compounds the carrier being a complex-forming compound able to form MRI-active complexes with paramagnetic metals the complex-forming compound being cyclic, e.g. DOTA
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
- G01N24/088—Assessment or manipulation of a chemical or biochemical reaction, e.g. verification whether a chemical reaction occurred or whether a ligand binds to a receptor in drug screening or assessing reaction kinetics
Abstract
The present invention provides pyrrolidines, salt, nuclear-magnetism probe and medicinal application, reagent, drug.This is supplied shown in benzamide methylpyrrole alkanes compound such as chemical structure of general formula (I), wherein, R1Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；R2Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；R3Selected from the alkyl or H that carbon atom number is 1~5；X1Selected from the alkylidene or missing that carbon atom number is 1~10；X2Selected from (CH2CH2O)n(wherein n=1~5) or missing；X3Selected from the alkyl that carbon atom number is 1~5；X4Selected from oxygen, sulphur, methylene；Ln is selected from Gd or Eu.The compound has and dopamine D2Receptor combines and relies on gadolinium, the europium chelate structure that the chelation group of intramolecular is formed, the specific Enhanced expressing dopamine D in Magnetic resonance imaging2The contrast of the tissue of receptor on the image is so as to be conducive to diagnose lesion type.
Description
Technical field
The present invention relates to benzamide methylpyrrole alkanes compound, pharmaceutically acceptable salt relaxes in nuclear magnetic resonance
Henan rate detection, Magnetic resonance imaging detection, the application of pituitary adenoma context of detection, the detection of NMR relaxation rate, nuclear magnetic resonance
Image checking, pituitary adenoma detection reagent, above-mentioned salt is in reagent, preparation treatment and the DOPA diagnosed to d2 dopamine receptor
Application in terms of the drug of the relevant disease of amine D2 expression of receptor, the reagent diagnosed to d2 dopamine receptor and treatment
The drug of relevant disease is expressed with d2 dopamine receptor.
Background technology
Dopamine (English name dopamine, hereinafter also referred to as DA) is most important catechu in central nervous system
Phenol amine neurotransmitters all act as highly important effect in maincenter and peripheral neverous system.The discovery of DA and its receptor
The important breakthrough of nerve, clinical psychiatry is caused.In the frontal lobe of normal pituitary and middle leaf texture, all there are certain levels
Dopamine D2Expression of receptor, they mediated dopamine to prolactin(PRL (English name prolactin, hereinafter also referred to as
) and the down regulation of the hormone secretions such as melanotropin PRL.Dopamine D2Receptor exists in PRL type pituitary adenomas
Height expression, in the treatment to PRL type pituitary adenomas, dopamine D2Receptor stimulating agent obtains good clinical effectiveness.At present
The therapy of most of prolactin(PRL type adenoma first choices is become.It is right with the development of molecular biology and Radiation Medicine technology
Dopamine D2The research of acceptor molecule achieves significant progress.Other than PRL type adenomas, in significant component of non-PRL types
Adenoma --- including in growth hormone type and the diseases such as promoting sexual gland hormone type and non-functional pituitary adenoma, all detecting dopamine
D2The expression of receptor, dopamine D2The therapeutic effect of receptor stimulating agent has obtained deep discussion and analysis.Dopamine D2Receptor is not
Expression with hypotype is also further detected, and the molecular mechanism to play a role also attracts attention.Dopamine D2By
The expression and dopamine D of body different subtype2Contact between receptor agonist treatment sensibility is worth further paying close attention to.Though
So in some specific results of study, also have differences and argue in report, but have reason to believe：With to dopamine D2
The constantly improve understood in depth and related drugs are developed of receptor and its hypotype molecular structure, dopamine D2Receptor stimulating agent
By in the treatment of different subtype pituitary adenoma, more positive effect is played.
In the prior art, exist and utilize dopamine D2Receptor it is this stress characteristic prevent and treat pituitary adenoma
The relevant technologies.However, in the prevention technique of the prior art, it is impossible to accurately detect dopamine D2The distribution density of receptor.
Invention content
Therefore, the technical problems to be solved by the invention are, provide a kind of benzamide methylpyrrole alkanes compound,
Its pharmaceutically acceptable salt, NMR relaxation rate detection, Magnetic resonance imaging detection, pituitary adenoma context of detection should
With and NMR relaxation rate detect, Magnetic resonance imaging detect, pituitary adenoma detection reagent.
In order to solve the above technical problem, the present invention provides following technical solutions.
The first aspect of the present invention provides a kind of benzamide methylpyrrole alkanes having shown in chemical structure of general formula (I)
Compound, the compound structure include three parts, are with the phenacyl pyrrole with d2 dopamine receptor binding function respectively
Cough up alkane part, with the octadentate ligand moiety of paramagnetic rare-earth metal ion-chelant and by above-mentioned two parts for example, by using alkynes-folded
The triazole linking arm part that nitrogen click chemistry method links together
Wherein,
R1Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R2Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R3Selected from the alkyl or H that carbon atom number is 1~5；
X1Selected from the alkylidene or missing that carbon atom number is 1~10；
X2Selected from-(CH2CH2O)n(wherein n=1~5) or missing；
X3Selected from the alkyl that carbon atom number is 1~5；
X4Selected from oxygen, sulphur, methylene；
Ln is selected from Gd or Eu.
Preferably, which has the structure shown in general formula (IA)
Preferably, which has the structure shown in general formula (IA-1)
Preferably, which has the structure shown in general formula (IA-1a)
Preferably, which has the structure shown in general formula (IA-1aa)
Preferably, which has the structure shown in general formula (IA-2)
Preferably, which has the structure shown in general formula (IA-2a)
Preferably, which has the structure shown in general formula (IA-2aa)
Preferably, which is any one in following compounds：
The compound has and dopamine D2Receptor combines and relies on gadolinium, the europium chelating knot that the chelation group of intramolecular is formed
Structure, the specific Enhanced expressing dopamine D in Magnetic resonance imaging2The contrast of the tissue of receptor on the image is so as to advantageous
In diagnosis lesion type.
In addition, the second aspect of the present invention provides the pharmaceutically acceptable salt of the compound.The medicine of the compound
Acceptable salt has and dopamine D on2Receptor combines and relies on gadolinium, the europium chelating knot that the chelation group of intramolecular is formed
Structure, the specific Enhanced expressing dopamine D in Magnetic resonance imaging2The contrast of the tissue of receptor on the image is so as to advantageous
In diagnosis lesion type.
In addition, the third aspect of the present invention provide the compound or the salt detect in NMR relaxation rate,
The application of Magnetic resonance imaging context of detection.
In addition, the fourth aspect of the present invention provides a kind of NMR relaxation rate detection, Magnetic resonance imaging detection reagent,
It includes the compound or the salt.
In addition, the third aspect of the present invention provide the compound or the salt detect in NMR relaxation rate,
Magnetic resonance imaging detection, the application of pituitary adenoma context of detection.
In addition, the fourth aspect of the present invention provides a kind of NMR relaxation rate detection, Magnetic resonance imaging detection, hypophysis
Adenoma detection reagent, it includes the compound or the salt.
In addition, the fifth aspect of the present invention provides a kind of compound or the salt to dopamine D2Receptor into
The reagent of row diagnosis prepares treatment and dopamine D2Application in terms of the drug of the relevant disease of expression of receptor.Preferably, it is described
With dopamine D2The relevant disease of expression of receptor includes but not limited to pituitary adenoma of different subtype etc..
In addition, the sixth aspect of the present invention offer is a kind of to dopamine D2The reagent that receptor is diagnosed, including described
Compound or the salt.
In addition, the seventh aspect of the present invention provides treatment and dopamine D2The drug of the relevant disease of expression of receptor, including
The compound or the salt.
The above compound of the present invention has low cytotoxicity.
Description of the drawings
Fig. 1 is the dose-effect relationship figure of Compound ira vitro cytotoxicity.
Fig. 2 is various concentration gradient dilution liquid embodiment compound 1 and Gd-DTPA contrast medium T1 weighted imaging figures.
Fig. 3 is poor in dopamine receptor expression for various concentration gradient dilution liquid embodiment compound 1 and Gd-DTPA contrast medium
Different cells in vitro T1 weighted imaging figures.
Fig. 4 is embodiment compound 1-5 and Gd-DTPA contrast medium in dopamine D2The cell body outer core of expression of receptor difference
Magnetic imaging T1 signal ratios.
Fig. 5 is embodiment compound 1 and Gd-DTPA contrast medium in dopamine D2NMR imaging in receptor knockout mice body.
Specific embodiment
Hereinafter, technical scheme of the present invention is described in detail.
The first aspect of the present invention provides a kind of benzamide methylpyrrole alkanes having shown in chemical structure of general formula (I)
Compound,
Wherein,
R1Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R2Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R3Selected from the alkyl or H that carbon atom number is 1~5；
X1Selected from the alkylidene or missing that carbon atom number is 1~10；
X2Selected from-(CH2CH2O)n(wherein n=1~5) or missing；
X3Selected from the alkyl that carbon atom number is 1~5；
X4Selected from oxygen, sulphur, methylene；
Ln is selected from Gd or Eu.
As can be seen that the compound of the present invention structure is made of three parts, it is to have to be combined with d2 dopamine receptor respectively
The phenacyl pyrrolidine moiety of function is matched with paramagnetism lanthanide rare metal ion Gd (gadolinium) or Eu (europium) octadentates chelated
Body portion and the triazole linking arm part that above-mentioned two parts link together for example, by using alkynes-nitrine click chemistry method.
For above-mentioned compound, there can also be stereoisomer, such as the compound has shown in general formula (IA)
Structure
More specifically, the compound has the structure shown in general formula (IA-1)
More specifically, the compound has the structure shown in general formula (IA-1a)
More specifically, the compound has the structure shown in general formula (IA-1aa)
More specifically, the compound has the structure shown in general formula (IA-2)
More specifically, the compound has the structure shown in general formula (IA-2a)
More specifically, the compound has the structure shown in general formula (IA-2aa)
More specifically, the compound is any one in following compounds：
The compound has and dopamine D2Receptor combines and relies on gadolinium, the europium chelating knot that the chelation group of intramolecular is formed
Structure, the specific Enhanced expressing dopamine D in Magnetic resonance imaging2The contrast of the tissue of receptor on the image is so as to advantageous
In diagnosis lesion type.
In addition, the second aspect of the present invention provides the pharmaceutically acceptable salt of the compound.The medicine of the compound
Acceptable salt has and dopamine D on2Receptor combines and relies on gadolinium, the europium chelating knot that the chelation group of intramolecular is formed
Structure, the specific Enhanced expressing dopamine D in Magnetic resonance imaging2The contrast of the tissue of receptor on the image is so as to advantageous
In diagnosis lesion type.
In addition, the third aspect of the present invention provide the compound or the salt detect in NMR relaxation rate,
Magnetic resonance imaging detection, the application of pituitary adenoma context of detection.
In addition, the fourth aspect of the present invention provides a kind of NMR relaxation rate detection, Magnetic resonance imaging detection, hypophysis
Adenoma detection reagent, it includes the compound or the salt.
In addition, the fifth aspect of the present invention provide a kind of compound or the salt to d2 dopamine receptor into
The reagent of row diagnosis prepares treatment and dopamine D2Application in terms of the drug of the relevant disease of expression of receptor.Preferably, it is described
With dopamine D2The relevant disease of expression of receptor includes but not limited to pituitary adenoma of different subtype etc..
In addition, the sixth aspect of the present invention offer is a kind of to dopamine D2The reagent that receptor is diagnosed, including described
Compound or the salt.
In addition, the seventh aspect of the present invention provides treatment and dopamine D2The drug of the relevant disease of expression of receptor, including
The compound or the salt.
It is illustrated below by way of specific embodiment.It should be noted that following embodiment is not limited to this hair
Bright protection domain.
Preparation example
The synthesis of each intermediate
The synthesis of preparation example 1.O- benzyl -3- butyne-1-ols
40ml anhydrous DMFs (dimethylformamide) are suspended from by the NaH (reagent concentration 60%) of 60% contents of 1.76g is molten
In, the lower addition 2.8g 3- butyne-1-ols of argon gas protection are bathed outside ice water, are instilled after the reaction was continued 30 minutes after the generation of no gas
5.24ml bromobenzyls drip off recession and remove ice-water bath, react at room temperature 12 hours, and 1mol/L aqueous hydrochloric acid solutions are added in after stopping reaction
200ml adds in EtOAc (ethyl acetate) 100ml and extracts 3 times, and organic layer is with 1mol/L aqueous hydrochloric acid solutions and saturation NaCl solution
Washing 3 times, with Na2SO4It is dry, filtrate is evaporated after filtering, the lower 80 DEG C of fractions of 4mmHg pressure are collected in vacuum distillation, obtain product
3.407g colorless oils, yield 53.2%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.32 (m, 5H), 4.44 (s, 2H), 3.52 (t, J=7.5Hz,
2H), 2.25 (td, J=7.5,3.0Hz, 2H), 2.02 (t, J=2.9Hz, 1H)
The synthesis of preparation example 2.O- benzyl -4- pentyne-1-alcohols
It is suspended from 1.76g 60%NaH are molten in 40ml anhydrous DMFs, bathes that argon gas protection is lower to add in 3.36g 3- penta outside ice water
Alkynes -1- alcohol instills 5.24ml bromobenzyls after the reaction was continued 30 minutes after the generation of no gas, drips off recession and remove ice-water bath, room temperature is anti-
Answer 12 hours, 1mol/L aqueous hydrochloric acid solution 200ml added in after stopping reaction, add in EtOAc 100ml and extract 3 times, organic layer with
1mol/L aqueous hydrochloric acid solutions and saturation NaCl solution are washed 3 times, with Na2SO4It is dry, filtrate is evaporated after filtering, vacuum distillation is received
Collect the lower 95 DEG C of fractions of 4mmHg pressure, obtain product 3.967g colorless oils, yield 57%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.31 (m, 5H), 4.47 (s, 2H), 3.43 (t, J=4.7Hz,
2H), 2.15 (td, J=8.0,3.1Hz, 2H), 1.92 (t, J=3.0Hz, 1H), 1.61 (tt, J=8.0,4.7Hz, 2H)
The synthesis of preparation example 3.O- benzyl -5- hexin -1- alcohol
Be suspended from 0.876g 60%NaH are molten in 20ml anhydrous DMFs, bathed outside ice water argon gas protection it is lower add in 1.95g 3- oneself
Alkynes -1- alcohol instills 2.6ml bromobenzyls after the reaction was continued 30 minutes after the generation of no gas, drips off recession and remove ice-water bath, reacts at room temperature
12 hours, 1mol/L aqueous hydrochloric acid solution 100ml are added in after stopping reaction, EtOAc 50ml is added in and extracts 3 times, organic layer with
1mol/L aqueous hydrochloric acid solutions and saturation NaCl solution are washed 3 times, with Na2SO4It is dry, filtrate is evaporated after filtering, vacuum distillation is received
Collect the lower 110 DEG C of fractions of 4mmHg pressure, obtain product 3.016g colorless oils, yield 80.6%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.33 (m, 5H), 4.42 (s, 2H), 3.54 (t, J=4.6Hz,
2H), 2.12 (td, J=7.8,3.0Hz, 2H), 1.88 (t, J=3.0Hz, 1H), 1.63 (m, 2H), 1.47 (t, J=7.8Hz,
2H).
The synthesis of preparation example 4.O- propargyls-diethylene glycol
It is suspended from 1.122g tBuOK (tertiary butyl oxygroup potassium) are molten in the anhydrous THF of 6.5ml (tetrahydrofuran), argon is bathed outside ice water
2.12g diethylene glycols are added under gas shielded, 1.18g propargyl bromides are instilled after being stirred to react 30 minutes, reacts 1 hour and removes after dripping off
Ice-water bath reacts at room temperature 12 hours, crosses diatomite filter after stopping reaction, filtrate obtains yellow oil after being evaporated, with 300-400
Preparative separation, mobile phase petroleum ether are pressed in mesh silica gel:Ethyl acetate=4:1, collect product frac after be evaporated product is faint yellow
Grease 0.976g, yield 72.8%.
Product analysis：1H NMR(400MHz,CDCl3) δ 4.24 (d, J=2.3Hz, 2H), 3.73 (m, 6H), 3.65 (m,
2H), 2.44 (t, J=2.3Hz, 1H), 2.13 (s, 1H)
The synthesis of preparation example 5.O- propargyls-O '-benzyl diethylene glycol
It is suspended from 0.8g 60%NaH are molten in 20ml anhydrous DMFs, bathes that argon gas protection is lower to add in 4 gainedization of preparation example outside ice water
Object O- propargyls-diethylene glycol 2.63g 3- butyne-1-ols are closed, are instilled after the reaction was continued 30 minutes after the generation of no gas
3.435g bromobenzyls drip off recession and remove ice-water bath, react at room temperature 12 hours, and 1mol/L aqueous hydrochloric acid solutions are added in after stopping reaction
100ml adds in EtOAc 50ml and extracts 3 times, and organic layer is washed 3 times with 1mol/L aqueous hydrochloric acid solutions and saturation NaCl solution, with
Na2SO4It is dry, filtrate is evaporated after filtering, to press preparative separation, mobile phase petroleum ether in 300-400 mesh silica gel:Ethyl acetate
=16:3, it is evaporated to obtain product colorless oil 1.295g, yield 30.3% after collecting product frac.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.32 (m, 5H), 4.43 (s, 2H), 4.11 (d, J=2.9Hz,
2H), 3.60 (m, 4H), 3.1 (t, J=2.9Hz, 1H)
The synthesis of preparation example 6.5- (4- benzyloxies butynyl) -2,3- dimethoxybenzoic acids
Bromo- 2, the 3- dimethoxybenzoic acids 0.531g (2.034mmol) of 5- are dissolved in 15ml diethylamine, add in 7.02mg
Pd(PPh3)2Cl2(Ph is phenyl) and 3.9mg CuI instill 0.488g preparation examples 1 after 80 DEG C of argon gas protection reflux 30 minutes and change
The 2ml diethylamine solutions of object O- benzyl -3- butyne-1-ols are closed, room temperature is stirred to react overnight after back flow reaction 5 hours, stops adding
Reaction solution is evaporated after heat, 80ml EtOAc are added in into gained brown residue, with saturated common salt water washing 80ml washings three times,
Organic layer is with Na2SO4It is dry, brownish red grease is evaporated to obtain after filtering, to press preparative separation, mobile phase in 300-400 mesh silica gel
Petroleum ether:Ethyl acetate=3:1, it collects target components and obtains yellow oil 0.382g, yield 55.3%
Product analysis：1H NMR(400MHz,CDCl3) δ 12.23 (s, 1H), 7.82 (d, J=1.9Hz, 1H), 7.65 (d, J
=1.9Hz, 1H), 7.30 (s, 5H), 4.47 (s, 2H), 3.91 (s, 3H), 3.79 (s, 3H), 3.54 (t, J=7.5Hz, 2H),
2.20 (t, J=7.5Hz, 2H).ESI-MS:Calculated value 341.14 [M+H]+, measured value：341.14.
The synthesis of preparation example 7.5- (5- benzyloxies pentynyl) -2,3- dimethoxybenzoic acids
Bromo- 2, the 3- dimethoxybenzoic acids 0.6g (2.3mmol) of 5- are dissolved in 15ml diethylamine, add in 8.4mg Pd
(PPh3)2Cl2With 4.4mg CuI, 2 compound O- benzyls -4- penta of 0.6g preparation examples is instilled after 80 DEG C of argon gas protection reflux 30 minutes
The 2ml diethylamine solutions of alkynes -1- alcohol, room temperature is stirred to react overnight after back flow reaction 5 hours, and reaction solution is evaporated after stopping heating,
80ml EtOAc are added in into gained brown residue, with saturated common salt water washing 80ml washings three times, organic layer is with Na2SO4It is dry
It is dry, brownish red grease is evaporated to obtain after filtering, to press preparative separation, mobile phase petroleum ether in 300-400 mesh silica gel:Ethyl acetate
=3:1, it collects target components and obtains yellow oil 0.382g, yield 55.3%
Product analysis：1H NMR(400MHz,CDCl3) δ 12.25 (s, 1H), 7.81 (d, J=1.9Hz, 1H), 7.62 (d, J
=1.9Hz, 1H), 7.32 (m, 5H), 4.47 (s, 2H), 3.95 (s, 3H), 3.79 (s, 3H), 3.53 (t, J=7.5Hz, 2H),
2.20 (t, J=7.5Hz, 2H).ESI-MS:Calculated value 355.15 [M+H]+, measured value：355.15.
The synthesis of preparation example 8.5- (6- benzyloxies hexin base) -2,3- dimethoxybenzoic acids
Bromo- 2, the 3- dimethoxybenzoic acids 0.6g (2.3mmol) of 5- are dissolved in 15ml diethylamine, add in 8.4mg Pd
(PPh3)2Cl2With 4.4mg CuI, 3 compound O- benzyls -5- of 0.649g preparation examples is instilled after 80 DEG C of argon gas protection reflux 30 minutes
The 2ml diethylamine solutions of hexin -1- alcohol, room temperature is stirred to react overnight after back flow reaction 5 hours, and reaction is evaporated after stopping heating
Liquid, into gained brown residue add in 80ml EtOAc, with saturated common salt water washing 80ml washing three times, organic layer with
Na2SO4It is dry, brownish red grease is evaporated to obtain after filtering, to press preparative separation, mobile phase petroleum ether in 300-400 mesh silica gel:
Ethyl acetate=4:1, it collects target components and obtains yellow oil 0.527g, yield 62.3%
Product analysis：1H NMR(400MHz,CDCl3) δ 11.72 (s, 1H), 7.85 (d, J=2.1Hz, 1H), 7.62 (d, J
=2.1Hz, 1H), 7.32 (m, 5H), 4.47 (s, 2H), 3.90 (s, 3H), 3.81 (s, 3H), 3.51 (t, J=7.5Hz, 2H),
2.18 (t, J=7.8Hz, 2H), 1.65 (q, J=7.7Hz, 2H), 1.44 (t, J=7.8Hz, 2H)；ESI-MS:Calculated value
369.16[M+H]+, measured value：369.16.
The synthesis of preparation example 9.5- (6- benzyloxies hexin base) -2,3- dimethoxybenzoic acids
Bromo- 2, the 3- dimethoxybenzoic acids 0.6g (2.3mmol) of 5- are dissolved in 15ml diethylamine, add in 8.4mg Pd
(PPh3)2Cl2With 4.4mg CuI, argon gas instilled after protecting 80 DEG C of reflux 30 minutes 5 compound O- propargyls of 0.807g preparation examples-
The 2ml diethylamine solutions of O '-benzyl diethylene glycol, room temperature is stirred to react overnight after back flow reaction 5 hours, is evaporated after stopping heating
Reaction solution, into gained brown residue add in 80ml EtOAc, with saturated common salt water washing 80ml washing three times, organic layer with
Na2SO4It is dry, brownish red grease is evaporated to obtain after filtering, to press preparative separation, mobile phase petroleum ether in 300-400 mesh silica gel:
Ethyl acetate=1:1, it collects target components and obtains yellow oil 0.187g, yield 19.6%
Product analysis：1H NMR(400MHz,CDCl3) δ 11.72 (s, 1H), 7.81 (d, J=1.9Hz, 1H), 7.62 (d, J
=1.9Hz, 1H), 7.35 (m, 5H), 4.47 (s, 2H), 4.12 (s, 2H), 3.90 (s, 3H), 3.79 (s, 3H), 3.60 (m,
8H).；ESI-MS:Calculated value 414.17 [M+H]+, measured value：414.17.
Preparation example 10.N- (5- (4- benzyloxies butynyl) -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (1b) synthesis
Compound 1a prepared by 0.51g (1.5mmol) preparation example 6 is dissolved in addition 0.39ml dichloros in 10ml dry toluenes
Sulfoxide, argon gas protect 60 DEG C of heating to be evaporated after reacting 1 hour, add in the anhydrous DCM of 10ml (dichloromethane), lower instillation is bathed outside ice water
(S)-2- amine methyl-1-N-ethyl pyrrole N is diluted after reaction being stirred at room temperature 8 hours with 50mlDCM, with saturation NaHCO3Aqueous solution,
Saturation NaCl aqueous solutions wash 3 times respectively, and organic layer is with Na2SO4Dry, filtering is evaporated to obtain yellowish grease, with silica gel medium pressure
Post separation, mobile phase petroleum ether:Ethyl acetate 2:1, it collects product frac merging and is evaporated to obtain yellowish grease 0.6g, yield
88.6%.
Product analysis：1H NMR(400MHz,CDCl3) δ 8.03 (d, J=2.1Hz, 1H), 7.51 (d, J=2.1Hz, 1H),
7.31 (m, 5H), 4.64 (d, J=12.3Hz, 1H), 4.36 (d, J=12.4Hz, 1H), 3.90 (s, 3H), 3.82 (s, 3H),
3.76 (m, 5H), 3.43 (dd, J=12.4,6.9Hz, 1H), 3.14 (m, 2H), 2.84 (m, 1H), 2.57 (m, 3H), 2.17 (m,
2H), 1.73 (m, 1H), 1.60 (m, 3H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:Calculated value 451.25 [M+H]+, actual measurement
Value：451.25.
Preparation example 11.N- (5- (5- benzyloxies pentynyl) -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (2b) synthesis
Compound 2a prepared by 0.53g (1.5mmol) preparation example 7 is dissolved in addition 0.39ml dichloros in 10ml dry toluenes
Sulfoxide, argon gas protect 60 DEG C of heating to be evaporated after reacting 1 hour, add in the anhydrous DCM of 10ml, and lower instillation (S) -2- amine first is bathed outside ice water
Base -1- N-ethyl pyrrole N 0.23g are diluted after reaction being stirred at room temperature 8 hours with 50mlDCM, with saturation NaHCO3Aqueous solution, saturation
NaCl aqueous solutions wash 3 times respectively, and organic layer is with Na2SO4Dry, filtering is evaporated to obtain yellowish grease, with silica gel medium pressure column point
From mobile phase petroleum ether:Ethyl acetate 3:1, it collects product frac merging and is evaporated to obtain yellowish grease 0.593g, yield
85.2%.
Product analysis：1H NMR(400MHz,CDCl3) δ 8.12 (d, J=1.9Hz, 1H), 7.54 (d, J=2.1Hz, 1H),
7.27 (m, 5H), 6.77 (s, 1H), 4.52 (d, J=12.3Hz, 1H), 4.31 (d, J=12.3Hz, 1H), 3.88 (d, J=
14.4Hz,6H),3.45(m,1H),3.18(m,4H),2.84(m,1H),2.59(m,1H),2.22(m,4H),1.95(m,2H),
1.77 (m, 1H), 1.60 (m, 3H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:Calculated value 465.27 [M+H]+, measured value：
465.27。
Preparation example 12.N- (5- (6- benzyloxies hexin base) -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (3b) synthesis
Compound 2a prepared by 0.53g (1.5mmol) preparation example 8 is dissolved in addition 0.39ml dichloros in 10ml dry toluenes
Sulfoxide, argon gas protect 60 DEG C of heating to be evaporated after reacting 1 hour, add in the anhydrous DCM of 10ml, and lower instillation (S) -2- amine first is bathed outside ice water
Base -1- N-ethyl pyrrole N 0.23g are diluted after reaction being stirred at room temperature 8 hours with 50mlDCM, with saturation NaHCO3Aqueous solution, saturation
NaCl aqueous solutions wash 3 times respectively, and organic layer is with Na2SO4Dry, filtering is evaporated to obtain yellowish grease, with silica gel medium pressure column point
From mobile phase petroleum ether:Ethyl acetate 3:1, it collects product frac merging and is evaporated to obtain yellowish grease 0.593g, yield
85.2%.
Product analysis：1H NMR(400MHz,CDCl3) δ 8.04 (d, J=2.1Hz, 1H), 7.45 (d, J=2.1Hz, 1H),
7.32 (m, 5H), 4.64 (d, J=12.3Hz, 1H), 4.31 (d, J=12.4Hz, 1H), 3.90 (s, 3H), 3.82 (s, 3H),
3.76 (m, 5H), 3.43 (dd, J=12.4,6.9Hz, 1H), 3.14 (m, 2H), 2.84 (dq, J=12.5,8.1Hz, 1H),
2.57 (m, 3H), 2.17 (m, 2H), 1.73 (m, 1H), 1.60 (m, 3H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:It calculates
Value 479.28 [M+H]+, measured value：479.28.
The synthesis of 13. compound 4b of preparation example
Compound 4a prepared by 0.414g (1mmol) preparation example 9 is dissolved in addition 0.39ml dichloros in 10ml dry toluenes
Sulfoxide, argon gas protect 60 DEG C of heating to be evaporated after reacting 1 hour, add in the anhydrous DCM of 10ml, and lower instillation (S) -2- amine first is bathed outside ice water
Base -1- N-ethyl pyrrole N 0.154g are diluted after reaction being stirred at room temperature 8 hours with 50ml EtOAc, with saturation NaHCO3Aqueous solution is satisfied
It is washed respectively 3 times with NaCl aqueous solutions, organic layer is with Na2SO4Dry, filtering is evaporated to obtain yellowish grease, with silica gel medium pressure column
Separation, mobile phase petroleum ether:Ethyl acetate 1:1 (2%TEA (triethylamine)) collects product frac merging and is evaporated to obtain yellowish oil
Shape object 0.396.g, yield 75.6%.
Product analysis：1H NMR(400MHz,CDCl3) δ 8.11 (d, J=1.9Hz, 1H), 7.55 (d, J=1.9Hz, 1H),
7.30 (m, 5H), 6.72 (s, 1H), 4.61 (d, J=12.4Hz, 1H), 4.21 (m, 2H), 4.05 (d, J=12.3Hz, 1H),
3.90 (s, 3H), 3.84 (s, 3H), 3.31 (m, 11H), 2.84 (dq, J=12.5,8.1Hz, 1H), 2.59 (p, J=6.9Hz,
1H), 2.17 (m, 2H), 1.75 (m, 1H), 1.60 (m, 3H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:Calculated value 479.28
[M+H]+, measured value：525.29.
Preparation example 14.N- (5- butanol base -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidine -2- bases) first
The synthesis of amine (1c)
Compound 1b prepared by 0.45g (1mmol) preparation example 10 is dissolved in 20ml methanol, adds in 45mg 10%Pd/
C, at room temperature catalytic hydrogenation, Hydrogen Vapor Pressure 50Psi, reaction filter after 4 hours, and filtrate is evaporated to obtain colorless oil 0.36g, yield
99%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.83 (d, J=1.9Hz, 1H), 7.24 (d, J=1.9Hz, 1H),
6.94 (s, 1H), 3.84 (d, J=8.4Hz, 6H), 3.43 (m, 3H), 3.11 (m, 3H), 2.84 (dq, J=12.5,8.1Hz,
1H), 2.62 (m, 2H), 2.17 (m, 2H), 1.64 (m, 8H), 1.24 (t, J=5.5Hz, 1H), 1.07 (t, J=8.0Hz,
3H).；ESI-MS:Calculated value 365.24 [M+H]+, measured value：365.24.
Preparation example 15.N- (5- amylalcohol base -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidine -2- bases) first
The synthesis of amine (2c)
Compound 2b prepared by 0.47g (1mmol) preparation example 11 is dissolved in 20ml methanol, adds in 45mg 10%Pd/
C, at room temperature catalytic hydrogenation, Hydrogen Vapor Pressure 50Psi, reaction filter after 4 hours, and filtrate is evaporated to obtain colorless oil 0.37g, yield
98%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.81 (d, J=1.9Hz, 1H), 7.27 (d, J=1.9Hz, 1H),
6.94 (s, 1H), 3.89 (d, J=8.4Hz, 6H), 3.42 (m, 3H), 3.11 (m, 3H), 2.84 (dq, J=12.5,8.1Hz,
1H), 2.62 (m, 2H), 2.12 (m, 2H), 1.64 (m, 8H), 1.24 (t, J=5.5Hz, 1H), 1.07 (t, J=8.0Hz,
3H).；ESI-MS:Calculated value 379.25 [M+H]+, measured value：379.25.
Preparation example 16.N- (5- hexanol base -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidine -2- bases) first
The synthesis of amine (3c)
Compound 3b prepared by 0.48g (1mmol) preparation example 12 is dissolved in 20ml methanol, adds in 45mg 10%Pd/
C, at room temperature catalytic hydrogenation, Hydrogen Vapor Pressure 50Psi, reaction filter after 4 hours, and filtrate is evaporated to obtain colorless oil 0.385g, production
Rate 98%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.54 (dt, J=2.2,1.2Hz, 1H), 6.47 (s, 1H), 3.93
(s,3H),3.78(s,3H),3.52(m,3H),3.16(m,2H),2.82(m,2H),2.63(m,2H),2.17(m,2H),1.63
(m, 9H), 1.31 (m, 5H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:Calculated value 393.27 [M+H]+, measured value：
393.27。
The synthesis of 17. compound 4c of preparation example
Compound 4b prepared by 0.52g (1mmol) preparation example 13 is dissolved in 20ml methanol, adds in 45mg 10%Pd/
C, at room temperature catalytic hydrogenation, Hydrogen Vapor Pressure 50Psi, reaction filter after 4 hours, and filtrate is evaporated to obtain colorless oil 0.416g, production
Rate 95%.
Product analysis：1H NMR(400MHz,CDCl3) δ 7.62 (dd, J=2.2,1.1Hz, 1H), 7.14 (dd, J=2.1,
1.1Hz,1H),6.77(s,1H),3.90(s,3H),3.81(s,3H),3.60(m,2H),3.36(m,3H),3.20(m,4H),
3.14 (m, 5H), 3.09 (m, 3H), 3.03 (m, 3H), 2.84 (dq, J=12.5,8.1Hz, 1H), 2.60 (m, 2H), 2.14 (m,
3H), 1.89 (m, 4H), 1.60 (m, 3H), 1.07 (t, J=8.0Hz, 3H)；ESI-MS:Calculated value 439.27 [M+H]+, actual measurement
Value：439.27.
Preparation example 18.N- (5- (O- methylsulfonyls) butanol base -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (1d) synthesis
Compound 1c prepared by 0.73g (2mmol) preparation example 14 is dissolved in 15ml DCM, adds in 0.84ml TEA,
The lower DCM liquid 2ml for instilling 0.31ml mesyl chlorides of argon gas protection are bathed outside ice water, reacts at room temperature 18 hours, is diluted with 80ml DCM
Reaction solution, with saturation NaHCO3With respectively washing 3 times of saturation NaCl aqueous solutions, organic layer is with Na2SO4Dry, filtrate concentrates after filtering
Yellow oil, compression leg preparative separation in silica gel, mobile phase petroleum ether:Ethyl acetate=1:2, it collects object fraction and obtains faint yellow oil
Shape object 0.734g, yield 83.2%.(MsCl is mesyl chloride)
Product analysis：ESI-MS:Calculated value 443.21 [M+H]+, measured value：443.21.
Preparation example 19.N- (5- (O- methylsulfonyls) amylalcohol base -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (2d) synthesis
Compound 2c prepared by 0.756g (2mmol) preparation example 15 is dissolved in 15ml DCM, adds in 0.84ml TEA,
The lower DCM liquid 2ml for instilling 0.31ml mesyl chlorides of argon gas protection are bathed outside ice water, reacts at room temperature 18 hours, is diluted with 80ml DCM
Reaction solution, with saturation NaHCO3With respectively washing 3 times of saturation NaCl aqueous solutions, organic layer is with Na2SO4Dry, filtrate concentrates after filtering
Yellow oil, compression leg preparative separation in silica gel, mobile phase petroleum ether:Ethyl acetate=1:2, it collects object fraction and obtains faint yellow oil
Shape object 0.781g, yield 85.6%.
Product analysis：ESI-MS:Calculated value 457.23 [M+H]+, measured value：457.23.
Preparation example 20.N- (5- (O- methylsulfonyls) hexanol base -2,3- Dimethoxybenzoyls)-(S)-(1- N-ethyl pyrrole Ns
Alkane -2- bases) methylamine (3d) synthesis
Compound 3c prepared by 0.784g (2mmol) preparation example 16 is dissolved in 15ml DCM, adds in 0.84ml TEA,
The lower DCM liquid 2ml for instilling 0.31ml mesyl chlorides of argon gas protection are bathed outside ice water, reacts at room temperature 18 hours, is diluted with 80ml DCM
Reaction solution, with saturation NaHCO3With respectively washing 3 times of saturation NaCl aqueous solutions, organic layer is with Na2SO4Dry, filtrate concentrates after filtering
Yellow oil, compression leg preparative separation in silica gel, mobile phase petroleum ether:Ethyl acetate=2:3, it collects object fraction and obtains faint yellow oil
Shape object 0.794g, yield 84.5%.
Product analysis：ESI-MS:Calculated value 471.25 [M+H]+, measured value：471.25.
The synthesis of 21. compound 4e of preparation example
Compound 4c prepared by 0.876g (2mmol) preparation example 17 is dissolved in 15ml DCM, adds in 0.84ml TEA,
The lower DCM liquid 2ml for instilling 0.31ml mesyl chlorides of argon gas protection are bathed outside ice water, reacts at room temperature 18 hours, is diluted with 80ml DCM
Reaction solution, with saturation NaHCO3With respectively washing 3 times of saturation NaCl aqueous solutions, organic layer is with Na2SO4Dry, filtrate concentrates after filtering
Yellow oil, compression leg preparative separation in silica gel, mobile phase petroleum ether:Ethyl acetate=1:3, collection object fraction is concentrated to give yellowish
Color grease 0.756g, yield 73.2%.
Product analysis：ESI-MS:Calculated value 517.25 [M+H]+, measured value：517.25.
Preparation example 22.N- (5- (4- nitrine butyl) -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidines -2-
Base) methylamine (1e) synthesis
Compound 1d prepared by 0.442g (1mmol) preparation example 18 is dissolved in 10ml DMF, adds in 0.195g nitrine
Change sodium, 45 DEG C of argon gas protection oil bath heating is reacted 8 hours, and room temperature reaction is stayed overnight, with 50ml EtOAc dilute reaction solutions, with saturation
NaCl aqueous solutions wash 3 times, and organic layer is with Na2SO4Dry, filtrate is concentrated to give water white transparency oily object 0.389g, yield after filtering
100%.
Product analysis：ESI-MS:Calculated value 389.24 [M]+, measured value：389.24.
Preparation example 23.N- (5- (5- nitrine amyl) -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidines -2-
Base) methylamine (2e) synthesis
Compound 2d prepared by 0.442g (1mmol) preparation example 19 is dissolved in 10ml DMF, adds in 0.195g nitrine
Change sodium, 45 DEG C of argon gas protection oil bath heating is reacted 8 hours, and room temperature reaction is stayed overnight, with 50ml EtOAc dilute reaction solutions, with saturation
NaCl aqueous solutions wash 3 times, and organic layer is with Na2SO4Dry, filtrate is concentrated to give water white transparency oily object 0.4g, yield after filtering
99%.
Product analysis：ESI-MS:Calculated value 403.52 [M]+, measured value：403.52.
Preparation example 24.N- (5- (6- nitrine hexyl) -2,3- Dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidines -2-
Base) methylamine (2e) synthesis
Compound 3d prepared by 0.442g (1mmol) preparation example 20 is dissolved in 10ml DMF, adds in 0.195g nitrine
Change sodium, 45 DEG C of argon gas protection oil bath heating is reacted 8 hours, and room temperature reaction is stayed overnight, with 50ml EtOAc dilute reaction solutions, with saturation
NaCl aqueous solutions wash 3 times, and organic layer is with Na2SO4Dry, filtrate is concentrated to give water white transparency oily object 0.42g, yield after filtering
100%.
Product analysis：ESI-MS:Calculated value 417.27 [M]+, measured value：417.27.
The synthesis of 25. compound 4e of preparation example
Compound 4d prepared by 0.516g (1mmol) preparation example 21 is dissolved in 10ml DMF, adds in 0.195g nitrine
Change sodium, 45 DEG C of argon gas protection oil bath heating is reacted 6 hours, and room temperature reaction is stayed overnight, with 50ml EtOAc dilute reaction solutions, with saturation
NaCl aqueous solutions wash 3 times, and organic layer is with Na2SO4Dry, filtrate is concentrated to give water white transparency oily object 0.42g, yield after filtering
100%.
Product analysis：ESI-MS:Calculated value 464.28 [M]+, measured value：464.28.
The preparation of preparation example 26.N- tert-butoxy carbonyls (O- benzyls) seryl diethylamine (6a)
By 11.814g (40mmol) N- tert-butoxy carbonyls (O- benzyls) serine (Boc-Ser (Bzl)-OH), 5.06g N-
HOSu NHS (HOSu) and 6.715gDCC (dicyclohexylcarbodiimide) are dissolved in the anhydrous DCM of 100ml, are bathed outside ice water
Lower addition DIEA (n,N-diisopropylethylamine) is restored room temperature after being stirred to react 30 minutes and is stirred to react 8 hours, filters reaction solution
By filtrate successively with 1mol/L hydrochloric acid, saturation NaHCO3Respectively washing 3 times of aqueous solution saturation NaCl aqueous solutions, organic layer is with Na2SO4
It is filtered after drying, 45 DEG C of vacuum drying of vacuum drying chamber, products therefrom is dissolved in the anhydrous DCM of 200ml after filtrate concentration, is instilled
It bathes and is stirred to react in 18.4ml ethylenediamine solutions, outside ice water, drip off recession and remove ice bath, react at room temperature 4 hours.Add in water 100ml
After be stirred to react 20 minutes, go to separatory funnel, organic phase retains, and water phase merges organic phase with Na after being extracted 3 times with DCM2SO4
It is filtered after drying, with silica gel medium pressure column chromatography separation mobile phase DCM after filtrate concentration:MeOH (methanol)=50:1, collect target
Brown oil 10.048g yields 74.5% are concentrated to give after fraction.
Product analysis：1H NMR(400MHz,CDCl3)δ7.32(m,5H),6.79(s,1H),6.24(s,1H),5.13
(t, J=7.0Hz, 1H), 4.91 (d, J=12.4Hz, 1H), 4.45 (d, J=12.3Hz, 1H), 4.19 (dd, J=12.5,
7.0Hz,1H),
3.78 (td, J=12.2,3.1Hz, 1H), 3.60 (dd, J=12.4,6.9Hz, 1H), 3.05 (td, J=12.1,
3.1Hz, 1H), 2.91 (m, 2H), 1.42 (d, J=18.4Hz, 11H)；ESI-MS:Calculated value 338.20 [M+H]+, measured value：
338.20。
The preparation of (O- benzyls) the seryl diethylamine hydrochloride of preparation example 27. (6b)
10ml methanol is added in compound 6a prepared by 9.418g (27.9mmol) preparation example 26, adds in 3mol/L saturations
The ethyl acetate solution of HCl, ice water bathe be stirred to react concentration of reaction solution after 3 hours outside, the drying 12 hours of 45 DEG C of vacuum drying chamber,
Faint yellow solid 8.118g yields 93.7%.
Product analysis：ESI-MS:Calculated value 238.15 [M+H]+, measured value：238.15.
The preparation of preparation example 28.2- methylol -4- aminoethyls ethylenediamines (6c)
Compound 6b prepared by 7.514g (21.67mmol) preparation example 27 is hanged and is dissolved in the anhydrous THF of 50ml, is added in
The borane dimethylsulfide ethereal solution 11ml of 9.89mmol/ml concentration, is added dropwise rear 65 DEG C of back flow reactions 36 hours, instills and adds in
6mol/L HCl/water solution 11ml, are heated to reflux rotary evaporated to dryness after 30 minutes, by above-mentioned product with ion exchange resin
Dowax50WX2 (a kind of title of strongly acidic styrene type cation exchange resin) purifies to obtain brown oil, by the grease
It adds in 100ml 6N HCl/water solution and concentrates reaction solution after the reaction 4 hours of 90 DEG C of tube sealing, it is brownly dry in vacuum drying chamber
Color grease obtains brown oil 3.34g, yield 63.4%.
Product analysis：1H NMR(400MHz,D2O) δ 3.86 (ddd, J=12.3,7.0,5.1Hz, 1H), 3.54 (ddd, J
=12.3,7.0,5.1Hz, 1H), 2.98 (m, 2H), 2.85 (m, 2H), 2.72 (m, 2H), 2.61 (dd, J=12.3,7.0Hz,
1H).。
Preparation example 29.N, N, N ', N ', N ",-five tert-butoxycarbonylmethyls of N "-(2- methylol -4- aminoethyls) ethylenediamine (6d)
Preparation
Compound 6c prepared by 3.34g (13.77mmol) preparation example 28 is dissolved in 40ml anhydrous DMFs, adds in DIEA
Reaction is stirred at room temperature under nitrogen protection in 36ml, dropwise addition bromo-acetic acid tert-butyl 15ml, after react at room temperature 16 hours, will react
Saturation NaCl aqueous solutions are added in after liquid concentration, are extracted 3 times with ethyl acetate, organic layer is with distilled water, saturation NaHCO3Aqueous solution,
Respectively washing 3 times of saturation NaCl aqueous solutions, Na2SO4It is dry, after filtering concentration residue is placed in brownly dry in vacuum drying chamber
Color grease obtains brown oil 6.555g, yield 67.7%.
Product analysis：ESI-MS:Calculated value 704.46 [M+H]+, measured value：704.46.
Preparation example 30.N, N, N ', N ', N ",-five tert-butoxycarbonylmethyls of N "-(2- methylol -4- aminoethyls) ethylenediamine (6e)
Preparation
Compound 6d prepared by 2.9g (4.1mmol) preparation example 29 is dissolved in 10ml anhydrous DMFs, argon gas protection is lower to be added in
60%NaH 0.656g, after being stirred to react 15 minutes, be added dropwise propargyl bromide 0.962ml, after react at room temperature 16 hours, addition
100mlDCM is washed 3 times with saturation NaCl aqueous solutions, and organic layer is with Na2SO4It is filtered after drying, filtrate is concentrated to give brownish red oil, with
Silica gel medium pressure prepares chromatographic isolation, mobile phase DCM:MeOH=20:1 separation, concentrates after collecting object fraction, obtains brown oil
Object 0.89g, yield 29.3%.,
Product analysis：ESI-MS:Calculated value 742.49 [M+H]+, measured value：742.49.
The synthesis of 31 compound 1f of preparation example
The compound 6E 0.8g prepared by preparation example 30 are dissolved in the change that 0.3g preparation examples 22 are added in 10ml ethyl alcohol and are prepared
The ethanol solution of object 1e is closed, adds 10ml distilled water, 13.7mg copper sulphate and 63.4mg sodium ascorbates is added in, is stirred to react 24
Hour, saturation NaCl aqueous solution 30ml are added in, with DCM extractions three times, merge organic layer with Na2SO4Dry, filtrate is dense after filtering
Standby silica gel post separation, mobile phase DCM are suppressed in contracting:MeOH=10:1, obtain 0.418g yellow wax after collecting the concentration of object fraction
Shape solid, yield 50%.
Product analysis：1H NMR(400MHz,CDCl3)δ8.81(s,1H),7.66(s,1H),7.40(s,1H),6.85(s,
1H), 4.60 (s, 2H), 4.33 (t, J=7.4Hz, 2H), 4.11 (s, 4H), 3.84 (s, 5H), 3.67 (d, J=4.4Hz, 2H),
3.43 (m, 10H), 3.00 (s, 2H), 2.78 (s, 6H), 2.57 (t, J=7.7Hz, 3H), 2.02 (m, 9H), 1.43 (d, J=
7.9Hz,45H).ESI-MS:Calculated value 1131.72 [M+H]+, measured value：1131.72.
The synthesis of 32 compound 1g of preparation example
Compound 1f 282mg prepared by preparation example 31 are dissolved in 15mlTFA (trifluoroacetic acid), it is small that reaction 2 is stirred at room temperature
When after concentration of reaction solution obtain brown oil, anhydrous ether is added to grind to obtain pale yellow powder 308mg, yield 100%.
Product analysis：ESI-MS:Calculated value 851.41 [M+H]+, measured value：851.41.
The synthesis of 33 compound 2f of preparation example
The compound 6e 0.8g prepared by preparation example 30 are dissolved in 10ml ethyl alcohol and add in prepared by 0.29g preparation examples 23
The ethanol solution of compound 2e adds 10ml distilled water, adds in 13.7mg copper sulphate and 63.4mg sodium ascorbates, is stirred to react
24 hours, saturation NaCl aqueous solution 30ml are added in, with DCM extractions three times, merge organic layer with Na2SO4It is dry, filtrate after filtering
Standby silica gel post separation, mobile phase DCM are suppressed in concentration:MeOH=10:1, obtain 0.428g yellow after collecting the concentration of object fraction
Waxy solid, yield 52%.
Product analysis：ESI-MS:Calculated value 1145.74 [M+H]+, measured value：1145.74.
The synthesis of 34 compound 2g of preparation example
Compound 2f 400mg prepared by preparation example 33 are dissolved in 15mlTFA, are concentrated after reaction being stirred at room temperature 2 hours anti-
Liquid is answered to obtain brown oil, anhydrous ether is added to grind to obtain pale yellow powder 436mg, yield 100%.
Product analysis：ESI-MS:Calculated value 865.42 [M+H]+, measured value：865.42.
The synthesis of 35 compound 3f of preparation example
The compound 6e 0.8g prepared by preparation example 30 are dissolved in the change that 0.3g preparation examples 24 are added in 10ml ethyl alcohol and are prepared
The ethanol solution of object 3e is closed, adds 10ml distilled water, 13.7mg copper sulphate and 63.4mg sodium ascorbates is added in, is stirred to react 24
Hour, saturation NaCl aqueous solution 30ml are added in, with DCM extractions three times, merge organic layer with Na2SO4Dry, filtrate is dense after filtering
Standby silica gel post separation, mobile phase DCM are suppressed in contracting:MeOH=15:1, obtain 0.459g yellow wax after collecting the concentration of object fraction
Shape solid, yield 55%.
Product analysis：ESI-MS:Calculated value 1159.75 [M+H]+, measured value：1159.75.
The synthesis of 36 compound 3g of preparation example
Compound 3f 400mg prepared by preparation example 35 are dissolved in 15ml TFA, are concentrated after reaction being stirred at room temperature 2 hours
Reaction solution obtains brown oil, and anhydrous ether is added to grind to obtain pale yellow powder 435mg, yield 100%.
Product analysis：ESI-MS:Calculated value 879.44 [M+H]+, measured value：879.44.
The synthesis of 37 compound 4f of preparation example
The compound 6e 0.8g prepared by preparation example 30 are dissolved in 10ml ethyl alcohol and add in prepared by 0.33g preparation examples 25
The ethanol solution of compound 3e adds 10ml distilled water, adds in 13.7mg copper sulphate and 63.4mg sodium ascorbates, is stirred to react
24 hours, saturation NaCl aqueous solution 30ml are added in, with DCM extractions three times, merge organic layer with Na2SO4It is dry, filtrate after filtering
Standby silica gel post separation, mobile phase DCM are suppressed in concentration:MeOH=15:1, obtain 0.548g whites after collecting the concentration of object fraction
Waxy solid, yield 63.2%.
Product analysis：ESI-MS:Calculated value 1205.76 [M+H]+, measured value：1205.76.
The synthesis of 38 compound 4g of preparation example
Compound 4f 450mg prepared by preparation example 37 are dissolved in 15ml TFA, are concentrated after reaction being stirred at room temperature 2 hours
Reaction solution obtains brown oil, and anhydrous ether is added to grind to obtain pale yellow powder 346mg, yield 100%.
Product analysis：ESI-MS:Calculated value 925.44 [M+H]+, measured value：925.44.
Preparation example 39N- (the bromo- 2,3- dimethoxybenzoyls of 5-)-(S)-(1- ethyl pyrrolidine -2- bases) methylamine (5a)
Synthesis
The bromo- 2,3- dimethoxybenzoic acids of 0.486g (1.86mmol) 5- are dissolved in 10ml dry toluenes and add in 0.39ml
Thionyl chloride, argon gas are evaporated after protecting 60 DEG C of heating reactions 1 hour, add in after 10ml anhydrous DCM bathed outside ice water it is lower instill (S)-
2- amine methyl-1-N-ethyl pyrrole N 0.286g is diluted after reaction being stirred at room temperature 8 hours with 50mlDCM, with saturation NaHCO3It is water-soluble
Liquid, saturation NaCl aqueous solutions wash 3 times respectively, and organic layer is with Na2SO4Dry, filtering is evaporated to obtain yellowish grease, with middle pressure
Silica gel post separation, mobile phase petroleum ether:Ethyl acetate 2:1, it collects product frac merging and is evaporated to obtain yellowish grease 1.628g,
Yield 87.5%.
Product analysis：1H NMR(400MHz,CDCl3) δ 8.05 (d, J=1.9Hz, 1H), 7.31 (d, J=2.0Hz, 1H),
6.52 (s, 1H), 3.90 (s, 3H), 3.82 (s, 3H), 3.43 (dd, J=12.4,6.9Hz, 1H), 3.13 (m, 2H), 2.84
(dq, J=12.5,8.1Hz, 1H), 2.59 (p, J=6.9Hz, 1H), 2.23 (m, 1H), 2.13 (dt, J=9.5,7.0Hz,
1H), 1.77 (m, 1H), 1.64 (pd, J=6.8,1.9Hz, 2H), 1.52 (dq, J=12.4,6.9Hz, 1H), 1.07 (t, J=
8.0Hz,3H；ESI-MS:Calculated value 371.09 [M+H]+, measured value：371.09.
Preparation example 40N- (5- nitrine -2,3- dimethoxybenzoyls)-(S)-(1- ethyl pyrrolidine -2- bases) methylamine (5b)
Synthesis
The 1.5g compounds 5a prepared by preparation example 39 is dissolved in 10ml dryings DMF, adds in 0.195g sodium azide,
0.06g CuI, 0.18g L-PROLINEs, 45 DEG C of argon gas protection oil bath heating are reacted 5 hours, and room temperature reaction is stayed overnight, with 50ml
EtOAc dilute reaction solutions, with saturation NaHCO3Aqueous solution, saturation NaCl aqueous solutions wash 3 times, and organic layer is with Na2SO4It is dry, mistake
Filtrate is concentrated to give yellow transparent grease 1.35g, yield 100% after filter.
Product analysis：ESI-MS:Calculated value 333.18 [M]+, measured value：333.18.
The synthesis of 41 compound 5f of preparation example
The compound 6e 0.8g prepared by preparation example 30 are dissolved in the change that 0.3g preparation examples 35 are added in 10ml ethyl alcohol and are prepared
The ethanol solution of object 5b is closed, adds 10ml distilled water, 13.7mg copper sulphate and 63.4mg sodium ascorbates is added in, is stirred to react 24
Hour, saturation NaCl aqueous solution 30ml are added in, with DCM extractions three times, merge organic layer with Na2SO4Dry, filtrate is dense after filtering
Standby silica gel post separation, mobile phase DCM are suppressed in contracting:MeOH=10:1, obtain 0.418g yellow oils after collecting the concentration of object fraction
Shape object, yield 50%.
Product analysis：1H NMR(400MHz,CDCl3)δ8.75(s,1H),7.54(s,1H),7.32(s,1H),6.77(s,
1H), 4.52 (s, 2H), 4.12 (t, J=7.4Hz, 2H), 4.02 (s, 4H), 3.81 (s, 5H), 3.57 (d, J=4.4Hz, 2H),
3.38 (m, 10H), 3.10 (s, 2H), 2.21 (m, 9H), 1.42 (d, J=7.9Hz, 45H) .ESI-MS:1075.66 [M of calculated value
+ H]+, measured value：1075.66.
The synthesis of 42 compound 5d of preparation example
Compound 5c 400mg prepared by preparation example 41 are dissolved in 15ml TFA, are concentrated after reaction being stirred at room temperature 2 hours
Reaction solution obtains brown oil, and anhydrous ether is added to grind to obtain pale yellow powder 438mg, yield 100%.
Product analysis：ESI-MS:Calculated value 795.34 [M+H]+, measured value：795.34.
Embodiment 1The synthesis of target compound 1H
The intermediate 1g 229mg (0.26mmol) prepared by preparation example 30 are dissolved in 5ml water, add in NaHCO3, adjust pH
To 6.5, ice water bathes the lower instillation 0.1g GdCl of stirring outside3Aqueous solution 5ml, remove ice bath after dripping off, pH=5.5, reaction 3 is small
When with 1mol/L NaOH tune pH to 8.5, add in acetone 200ml, pH kept to generate white precipitate and filter in vain in 7.5-8.0
Color solid 165mg, yield 62.3%.
Product analysis：ESI-MS:1004.33 [M-H of calculated value2O+H]+, measured value：1004.33, molecular ion peak is shown
For gadolinium isotope characteristic signal.
Embodiment 2The synthesis of target compound 2H
The intermediate 2g 325mg (0.26mmol) prepared by preparation example 32 are dissolved in 5ml water, add in NaHCO3, adjust pH
To 6.5, ice water bathes the lower instillation 0.1g GdCl of stirring outside3Aqueous solution 5ml, remove ice bath after dripping off, pH=5.5, reaction 3 is small
When with 1mol/L NaOH tune pH to 8.5, add in acetone 200ml, pH kept to generate white precipitate and filter in vain in 7.5-8.0
Color solid 178mg, yield 66.2%.
Product analysis：ESI-MS:1018.32 [M-H of calculated value2O+H]+, measured value：1018.32, molecular ion peak is shown
For gadolinium isotope characteristic signal.
Embodiment 3The synthesis of target compound 3H
The intermediate 3g 229mg (0.18mmol) prepared by preparation example 34 are dissolved in 5ml water, add in NaHCO3, adjust pH
To 6.5, ice water bathes the lower instillation 0.1g GdCl of stirring outside3Aqueous solution 5ml, remove ice bath after dripping off, pH=5.5, reaction 3 is small
When with 1mol/L NaOH tune pH to 8.5, add in acetone 200ml, pH kept to generate white precipitate and filter in vain in 7.5-8.0
Color solid 168mg, yield 89%.
Product analysis：ESI-MS:1032.24 [M-H of calculated value2O+H]+, measured value：1032.24, molecular ion peak is shown
For gadolinium isotope characteristic signal.
Embodiment 4The synthesis of target compound 4H
The intermediate 4g 229mg (0.18mmol) prepared by preparation example 36 are dissolved in 5ml water, add in NaHCO3, adjust pH
To 6.5, ice water bathes the lower instillation 0.1g GdCl of stirring outside3Aqueous solution 5ml, remove ice bath after dripping off, pH=5.5, reaction 3 is small
When with 1mol/L NaOH tune pH to 8.5, add in acetone 200ml, pH kept to generate white precipitate and filter in vain in 7.5-8.0
Color solid 168mg, yield 89%.
Product analysis：ESI-MS:1032.24 [M-H of calculated value2O+H]+, measured value：1032.24, molecular ion peak is shown
For gadolinium isotope characteristic signal.
Embodiment 5The synthesis of target compound 5H
The intermediate 5d 143mg (0.18mmol) prepared by preparation example 42 are dissolved in 5ml water, add in NaHCO3, adjust pH
To 6.5, ice water bathes the lower instillation 0.1g GdCl of stirring outside3Aqueous solution 5ml, remove ice bath after dripping off, pH=5.5, reaction 3 is small
When with 1mol/L NaOH tune pH to 8.5, add in acetone 200ml, pH kept to generate white precipitate and filter in vain in 7.5-8.0
Color solid 156mg, yield 90.2%.
Product analysis：ESI-MS:948.24 [M-H of calculated value2O+H]+, measured value：948.24 molecular ion peak is shown as
Gadolinium isotope characteristic signal.
It should be noted that the compound of the present invention can also be obtained by other organic reaction routes, the skill of this field
Art personnel on the basis of the disclosure of invention even on the basis of technique leading, can be by designing organic reaction road
Line and obtain the compound of the present invention.
The biological activity experimental sections of target compound
1. embodiment Compound ira vitro cytotoxicity screening technique of experimental example
Cell and reagent：Rat pituitary oncocyte system GH3 is U.S.'s ATCC Products.DMEM culture mediums and high-quality tire ox
Serum is purchased from Gibco companies, and dimethyl sulfoxide (DMSO) (Dimethylsulphoxide, DMSO), trypsase is purchased from U.S. Sigma public affairs
Department, MTT are purchased from Genview companies of the U.S., and purchased from North China drugmaker, other reagents are commercially available analysis for penicillin, streptomysin
It is pure.
Laboratory apparatus and equipment：HERAcell150 types CO2Cell incubator (congratulates Li Shi companies, Germany), and IMT-2 types fall
Put microscope (Olympus Corp, Japan), 550 type enzyme-linked immunosorbent assay instruments (BIO-RAD companies, the U.S.)；Consumptive material is culture
Ware, 96 cell well culture plates (Costar companies, the U.S.) etc..
Body outer screening test method
1. routine culture and the passage of pituitary adenoma cells GH3
The rat pituitary frozen oncocyte system GH3 cells is taken to recover, with fetal calf serum containing 100mL/L and 100U/mL
Penicillin, 100U/mL streptomysins DMEM culture mediums in 37 DEG C, 5%CO2And it is trained in the constant incubator of saturated humidity condition
It supports, is periodically passed on according to growing state, tested using the cell after the 3rd passage.
2. influence of the embodiment compound to pituitary adenoma GH3 cell Proliferations
It it is 6 groups by GH3 points of the pituitary adenoma of the 3rd passage, experimental group is embodiment compound 1-5, with containing 10% tire ox
The DMEM culture mediums of serum are configured to the ultimate density of 5,10,50,500,5000 μM/L, and blank control group (blank group) only adds
Enter to contain the DMEM culture mediums of 10% fetal calf serum, it is thin that passage cell by every 4000/100ul in hole is inoculated in 3 96 holes respectively
In born of the same parents' culture plate, it is divided into 5 groups by above-mentioned grouping scheme, every group sets 8 repetitions.Liquid is changed after cell is adherent for 24 hours, is separately added into
The compound for stating respective concentration is placed in 37 DEG C, 5%CO2And it is cultivated in saturated humidity condition constant incubator.After 72 hours, divide
(MTT is 3- (4,5- dimethyl thiophenes to the MTT solution of 20 a concentration of 5mg/ml of μ l of every hole addition not in the culture hole of required detection
Azoles -2) -2,5- diphenyltetrazolium bromide bromides, trade name tetrazolium bromide, English for 3- (4,5-dimethyl-2-thiazolyl) -
2,5-diphenyl-2-H-tetrazolium bromide, solvent are dimethyl sulfoxide (DMSO), i.e. DMSO), 37 DEG C are continued to be incubated
4h terminates culture, careful to draw culture supernatant in hole, and 100 μ l dimethyl sulfoxide (DMSO)s (DMSO) are added in per hole, vibrate 10min.
490nm wavelength is selected, each hole absorbance value OD is measured on enzyme-linked immunosorbent assay instrument490。
It is the dose-effect relationship figure of Compound ira vitro cytotoxicity as shown in Figure 1.As can be seen that test-compound is in 5-5000
μM/L concentration under pituitary adenoma cells proliferation is had no significant effect, this series compound have low cytotoxicity.Thus may be used
Know, the benzamide methylpyrrole alkanes compound that the present invention synthesizes is used for the nuclear magnetic resonance of pituitary adenoma cells as contrast agent
Imaging has hypotoxicity.
2. embodiment compound NMR relaxation rate of experimental example detects
(Gd-DTPA, gadolinium diethylene triaminepenta acetate face for the identical embodiment compound 1-5 of compound concentration and Magnevist Solution
Bed nuclear magnetism contrast agent) contrast medium, embodiment compound 1-5 and Gd-DTPA are used into PBS (phosphate buffer, wherein phosphorus respectively
A concentration of 0.01mol/L, pH 7.2 of acid buffering solution) to be configured to 0,5 μM, 10 μM, 50 μM, 500 μM and 5mM finally dense
Degree is resuspended in 0.5mL microcentrifugal tubes (it is managed for Eppendof, hereinafter also referred to as Eppendof pipes).By Eppendof
Pipe is put into progress magnetic resonance imaging T1 weighted imagings, Fig. 2 on Bruker toys nuclear magnetic resonance machine (German Bruker companies)
Various concentration gradient dilution liquid embodiment compound 1 and Gd-DTPA contrast medium T1 weighted imaging figures.Measure embodiment compound and
Gd-DTPA contrast medium signals select region of interest (ROI, return on of the same size in every group of sample size respectively
Interesting) measuring signal intensity takes the average value of measurement result.Experiment is repeated 3 times.And by embodiment compound 1-5 and
The signal strength of Gd-DTPA contrast medium carries out correlation regression line analysis, and slope is that the relaxation rate (R) of contrast medium (is shown in Table
1).Ratio, that is, signal difference of the relaxation rate of any compound and contrast medium in embodiment compound 1-5, as shown in table 1, R '
=RCompound/RGd-DTPA, RCompound 1/RGd-DTPA=1.15, RCompound 2/RGd-DTPA=1.13, RCompound 3/RGd-DTPA=1.14, RCompound 4/
RGd-DTPA=1.12, RCompound 5/RGd-DTPA=1.16, as a result show any compound/Gd-DTPA in embodiment compound 1-5
Signal ratio 1 or so, should the result shows that, embodiment compound 1-5 and the magnetic resonance under same concentrations of Gd-DTPA contrast medium
T1 imagings effect is close, and signal difference and relaxation rate difference are similar.I.e. at normal temperatures, embodiment compound 1-5 is the same as Gd-DTPA pairs
It is more the same than agent that all there are preferable nuclear-magnetism imaging results.It further demonstrates that, the benzamide crassitude that the present invention synthesizes
Class compound has good contrast ability, can be as the contrast agent of Magnetic Resonance Imaging.
It should be noted that Magnevist Solution (Gd-DTPA) be a kind of current commonly used small molecular core magnetic resonance or
Magnetic resonance contrast agent is a kind of non-specific contrast medium of non-selectivity.
1 embodiment compound of table and Gd-DTPA contrast medium relaxation rate ratios
3. embodiment compound of experimental example is detected in the cells in vitro Magnetic resonance imaging of dopamine receptor differential expression
By dopamine D2The few pituitary adenoma GH3 cells of expression of receptor amount (deriving from ATCC companies) and dopamine D2Receptor
PC12 cells (derive from ATCC companies) more than expression quantity respectively with fetal calf serum containing 100mL/L and 100U/mL penicillin,
The DMEM culture mediums of 100U/mL streptomysins are in 37 DEG C, 5%CO2And cultivated in the constant incubator of saturated humidity condition, according to life
Long situation periodically passes on, and is tested using the cell after the 3rd passage.By GH3 and PC12 cells respectively with 1*106A/ml
Cell density kind in six orifice plates, after cell is adherent after by embodiment compound 1-5 and the Gd-DTPA contrast medium of various concentration
(0,5 μM, 10 μM, 50 μM, 500 μM and 5mM, solvent are the PBS of concentration 0.01mol/L, pH 7.2) is separately added into GH3 and PC12
4h is incubated in cell culture medium, is then rinsed 3 times with above-mentioned PBS, with the complete vitellophag of 0.25% trypsase,
1200rpm centrifuges 4min, is resuspended in 0.5mL Eppendof pipes.Eppendof pipes are put into Bruker toy nuclear magnetic resonance
Magnetic resonance imaging T1 weighted imagings are carried out on machine (German Bruker companies), Fig. 3 is various concentration gradient dilution liquid embodiment
Object 1 and Gd-DTPA contrast medium are closed in dopamine D2T1 weighted imagings in GH3 the and PC12 cells of expression of receptor situation difference
Figure.As shown in figure 3, in figure 3 a, Gd-DTPA contrast medium is in dopamine D2The GH3 and PC12 that expression of receptor situation has differences
Cell center magnetic signal strength no significant difference, and in figure 3b, embodiment compound 1 is containing dopamine D2The cell of receptor
The middle nuclear magnetic signal Strength Changes that concentration gradient is presented and relies on, while in dopamine D2In the higher PC12 cells of expression of receptor amount
Nuclear magnetic signal intensity is significantly higher than in dopamine D2The relatively low GH3 cell center magnetic signal strengths of expression of receptor amount.It measures and implements
Example compound 1-5 and Gd-DTPA contrast medium signal selects region of interest ROI of the same size in every group of sample size to measure respectively
Signal strength takes the average value of measurement result, and the concentration of wherein embodiment compound 1-5 and Gd-DTPA contrast medium is 5mM.
Experiment is repeated 3 times.The results are shown in Figure 4, and Gd-DTPA is in dopamine D2The GH3 and PC12 that expression of receptor situation has differences are thin
Nuclear magnetic signal ratio no significant difference in born of the same parents, and 5mM embodiment compound 1-5 are containing dopamine D2The cell center of receptor
Magnetic signal strength is significantly higher than Gd-DTPA contrast medium, while in dopamine D2Nuclear-magnetism in the higher PC12 cells of expression of receptor amount
Signal strength is significantly higher than in dopamine D2The relatively low GH3 cell center magnetic signal strengths of expression of receptor amount.The result is prompted, phase
Compared with Gd-DTPA contrast medium, in vitro on cellular level, embodiment compound 1-5 has dopamine D2Receptor target, can be with
It targets nuclear-magnetism and shows dopamine D2Acceptor levels.
4. embodiment compound of experimental example nuclear-magnetism Magnetic resonance imaging in dopamine knock-out mice body detects
Dopamine D2Receptor knockout mice is given by Chinese Academy of Sciences's Shanghai Neuroscience Research.By wild-type mice (DOPA
Amine D2Receptor normal expression) and dopamine D2Receptor knockout mice is divided into two groups, is respectively placed in Bruker toy nuclear magnetic resonance machines
Progress magnetic resonance imaging imaging in (German Bruker companies), t1 weighted image before acquisition hypophysis developer is strengthened, and respectively at
Tail vein injection embodiment compound 1 and Gd-DTPA contrast medium (dosage is 0.5mmol/kg with the batheroom scale of mouse),
After 10min, the mouse for injecting embodiment compound 1 and Gd-DTPA contrast medium is placed in Bruker toy nuclear magnetic resonance machine (morals
Bruker companies of state) on carry out magnetic resonance imaging imaging respectively, obtain t1 weighted image after hypophysis developer is strengthened, image results
See Fig. 5.Before injection, the hypophysis of wild-type mice (Fig. 5 A, 5E) and dopamine knock-out mice (Fig. 5 C, 5G) is shown in T1 weightings
For low signal, after injection after embodiment compound 1, wild-type mice hypophysis position signal occurs being remarkably reinforced (Fig. 5 B), and
Dopamine D2There is (Fig. 5 D) without enhancing signal in receptor knockout mice hypophysis position.After injection after Gd-DTPA contrast medium, wild type
Mouse Pituitary position (Fig. 5 F) and dopamine D2Receptor knockout mice hypophysis position (Fig. 5 H) signal is remarkably reinforced.The knot
Fruit shows, compared to Gd-DTPA contrast medium, to confirm that embodiment compound has dopamine D in animal level in vivo2Receptor target
Tropism can target nuclear-magnetism and show dopamine D2Acceptor levels.
In conjunction with the embodiments 3 and embodiment 4 experimental configuration it is found that the benzamide methylpyrrole alkanes that synthesizes of the present invention
Close object and its pharmaceutically acceptable salt, can specificity and dopamine D2Receptor combines, that is to say, that benzamide methyl pyrrole
Alkyl compound and its pharmaceutically acceptable salt are coughed up, it can targets identification dopamine D2Receptor.It further illustrates, benzamide first
Base pyrrolidines and its pharmaceutically acceptable salt can be applied to pituitary adenoma magnetic resonance detection or nuclear magnetic resonance inspection
In survey.
Although describing technical scheme of the present invention with reference to certain embodiments, those skilled in the art will manage
Solution, without departing from the present invention, can carry out a variety of variations.These variations should be considered as without departing from the present invention
Protection domain, and protection scope of the present invention is subject to the content of claims.
Claims (14)
1. with the benzamide methylpyrrole alkanes compound shown in chemical structure of general formula (I),
Wherein,
R1Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R2Selected from methyl, ethyl, methoxyl group, ethyoxyl, halogen；
R3Selected from the alkyl or H that carbon atom number is 1~5；
X1Selected from the alkylidene or missing that carbon atom number is 1~10；
X2Selected from-(CH2CH2O)n, wherein n=1~5 or missing；
X3Selected from the alkyl that carbon atom number is 1~5；
X4Selected from oxygen, sulphur, methylene；
Ln is selected from Gd or Eu.
2. compound according to claim 1, with the structure shown in general formula (IA)
3. compound according to claim 2, with the structure shown in general formula (IA-1)
4. compound according to claim 3, with the structure shown in general formula (IA-1a)
5. compound according to claim 4, with the structure shown in general formula (IA-1aa)
6. compound according to claim 2, with the structure shown in general formula (IA-2)
7. compound according to claim 6, with the structure shown in general formula (IA-2a)
8. compound according to claim 7, with the structure shown in general formula (IA-2aa)
9. compound according to claim 1, for any one in following compounds：
10. the pharmaceutically acceptable salt of compound according to any one of claims 1 to 9.
11. compound according to any one of claims 1 to 9 or salt according to any one of claims 10 are being prepared for nuclear magnetic resonance
Application in the nuclear-magnetism probe that relaxation rate detection, Magnetic resonance imaging detection, pituitary adenoma detect.
12. a kind of NMR relaxation rate detection, Magnetic resonance imaging detection, pituitary adenoma detection reagent, will it includes right
Ask the compound described in any one of 1~9 or salt according to any one of claims 10.
13. compound according to any one of claims 1 to 9 or salt according to any one of claims 10 are being prepared to dopamine D2By
Application in the reagent that body is diagnosed.
It is 14. a kind of to dopamine D2The reagent that receptor is diagnosed, including compound according to any one of claims 1 to 9 or
Salt according to any one of claims 10.
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