CN107216352A - Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application - Google Patents
Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application Download PDFInfo
- Publication number
- CN107216352A CN107216352A CN201710436442.4A CN201710436442A CN107216352A CN 107216352 A CN107216352 A CN 107216352A CN 201710436442 A CN201710436442 A CN 201710436442A CN 107216352 A CN107216352 A CN 107216352A
- Authority
- CN
- China
- Prior art keywords
- pyridine derivative
- mitochondrially targeted
- dihydrogen pyridine
- preparation
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
- C07F9/576—Six-membered rings
- C07F9/59—Hydrogenated pyridine rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
- C07F9/65586—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system at least one of the hetero rings does not contain nitrogen as ring hetero atom
Abstract
The invention discloses the preparation method and application of Mitochondrially targeted dihydropyridine compounds, Mitochondrially targeted dihydropyridine compounds have structure shown in formula (I) and (II)：
Description
Technical field
The present invention relates to Mitochondrially targeted dihydrogen pyridine derivative and Preparation method and use, belong to field of medicaments.
Background technology
In recent years, due to the fast development of nuclear science and radiating medical, life of the ionising radiation from us is more and more nearer, by
To the risk also more and more higher of ionization radiation injury, the mistake of core relevant staff, medical inspection and treatment is especially engaged in
Patient in journey, the excessive ray due to touching and the ionization radiation injury for causing body, cause body normal function to be lacked of proper care, body
Internal oxidition stress, or even induce many chronic diseases and ultimately result in aging and death.Ionization radiation injury is mainly high-energy ray
The damage that body produces function macromolecular is penetrated, its harm is divided into coup injury and indirect injury, and coup injury is due to big agent
Amount ray is directly acted in the eucaryotic cell structure in body, causes the DNA damage of nucleus, induces body oxidative stress and each
Class chronic disease；Indirect injury is also ionization common during we live as a kind of main damage of effect of low dose radiation
Damage, its principle is because high-energy ray ionizes internal water and oxygen, a large amount of free radicals of generation, the excessive freedom in this part
Base cannot effectively remove the macro-molecular protein that will be damaged in human body, lipid and nucleic acid, these macromoleculars biology work(
The obstacle or missing of energy can cause body dysfunction or disease.Ionising radiation be because the intracellular water of ray ionization simultaneously
Free radical is produced, radical damage large biological molecule causes ionization radiation injury, still, the radical life pole that ionizing water is produced
Its is of short duration, will be disappeared after a few minutes, and most continuation ionization damages rely primarily on internal delay active oxygen radical
Generation so as to causing internal functional molecular to be persistently damaged, cause response to oxidative stress.
Mitochondria, as the place of intracellular oxidation breathing, is also the intracellular only possible subcellular fraction for producing free radical
Structure, research shows that ionising radiation causes structure of mitochondria dysfunction, and continual generation free radical is produced to cell
Damage.And mitochondrial function damage can also produce oneself same caused by aging or due to extraneous some chemical factors
By base, therefore mitochondria is protected not to be damaged and remove the Intramitochondrial free radical of damage, aging as mitochondrial function barrier
Hinder the top priority of disease.
At present, dihydropyridine compounds, because its hypotoxicity has clinically been used as Ca2+Antagonist is used as drop
There is nicotinamide-adenine dinucleotide phosphate (NADPH) in pressing, organism, be used as a kind of important dihydropyridine endogenous
Antioxidant, can remove the oxidation materials such as endogenic free radical, but the particularity of NADPH structures can not efficiently gather
Collect the active oxygen radical removed in mitochondria in mitochondria, therefore, effectively performance can be gathered in mitochondria clearly by needing badly
Except the effect of free radical, mitochondria is assisted to remove the compound of the free radical of excess generation.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art there is provided Mitochondrially targeted dihydrogen pyridine derivative.
Second object of the present invention is to provide the preparation method of Mitochondrially targeted dihydrogen pyridine derivative.
Third object of the present invention is to provide the application of Mitochondrially targeted dihydrogen pyridine derivative.
Technical scheme is summarized as follows：
Mitochondrially targeted dihydrogen pyridine derivative, with following structure
Wherein：R is hydrogen atom, phenyl ring or thiophene；X is chlorine, bromine or iodine.
The preparation method of Mitochondrially targeted dihydrogen pyridine derivative (I), comprises the following steps：
1) by triphenylphosphine and 3- halogen propyl alcohol reaction generation (3- hydroxypropyls) triphenyl phosphonium halides；
2) by ammonium hydrogen carbonate, ethyl acetoacetate withThe back flow reaction in ethanol water, obtains compound 7, changes
Compound 7 obtains compound 8 by sodium hydrate aqueous solution hydrolysis, compound 8 with step 1) (3- hydroxypropyls) triphen for obtaining
Base phosphorus Halides is anti-under 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate and I-hydroxybenzotriazole catalysis
Should, obtain the dihydrogen pyridine derivative shown in Formulas I；
Reaction equation is：
Wherein：R is hydrogen atom, phenyl ring or thiophene；X is chlorine, bromine or iodine.
The preparation method of Mitochondrially targeted dihydrogen pyridine derivative (II), comprises the following steps：
By triphenylphosphine and 3- halogen propyl alcohol reaction generation (3- hydroxypropyls) triphenyl phosphonium halides；By (3- hydroxypropyls) triphenyl
Phosphorus Halides obtains (3- bromopropyls) triphenyl phosphonium halides with HBr reactions；(3- bromopropyls) triphenyl phosphonium halides and niacinamide are anti-
Compound 14 should be obtained, under conditions of pH=8~9, by sodium hydrosulfite reducing compound 14, the dihydro pyrrole shown in formula II is obtained
Piperidine derivatives；
Reaction equation is：
Wherein：X is chlorine, bromine or iodine.
Above-mentioned Mitochondrially targeted dihydrogen pyridine derivative is preparing the application for the treatment of alzheimer medicine.
Above-mentioned Mitochondrially targeted dihydrogen pyridine derivative is preparing the application for the treatment of Parkinsonian drugs.
Above-mentioned Mitochondrially targeted dihydrogen pyridine derivative is preparing the application of radioprotector.
Advantages of the present invention：
It is demonstrated experimentally that the Mitochondrially targeted dihydrogen pyridine derivative of the present invention can effectively be gathered in mitochondria and remove electricity
From the active oxygen (ROS) produced by radiation, therefore the relevant disease caused by free radical, such as alzheimer disease and ionization are damaged
There is purposes in terms of hindering disease treatment.This preparation method is simple, and product is easy to get, pollution-free.
Brief description of the drawings
Fig. 1 is that Mitochondrially targeted dihydrogen pyridine derivative can substantially increase the survival rate of cell after irradiation.
Fig. 2 is that Mitochondrially targeted dihydrogen pyridine derivative Scavenging ability substantially increases.
Fig. 3 is that Mitochondrially targeted dihydrogen pyridine derivative protection DNA double chain ability increased.
Fig. 4 can effectively be gathered in mitochondria for Mitochondrially targeted dihydrogen pyridine derivative and play a role.
The Mitochondrially targeted dihydrogen pyridine derivatives of 1- (I -1) in Fig. 4；The Mitochondrially targeted dihydrogen pyridine derivatives of 2- (I-
2)；The Mitochondrially targeted dihydrogen pyridine derivatives of 3- (I -3)；The Mitochondrially targeted dihydrogen pyridine derivatives of 4- (II).
Embodiment
Determining instrument：Nuclear magnetic resonance VARIAN INOVA 500MHz type NMRs.Mass spectrum Waters 3100
Level Four bar mass spectrograph.
With reference to specific embodiment, the present invention is further illustrated, and embodiments of the invention are in order that this area
Technical staff better understood when the present invention, but the present invention is not imposed any restrictions.
Embodiment 1
The preparation method of Mitochondrially targeted dihydrogen pyridine derivative (I-1), comprises the following steps：
(1) triphenylphosphine (9.4g, 35.9mmol) is sequentially added in the 50ml single port vials equipped with spherical condensation tube,
Bromo- 1 propyl alcohol (0.5g, 3.6mmol) of 3- and 10ml DMFs (DMF), 100 DEG C are stirred at reflux 10h, reaction knot
Shu Houyong is cooled to the solid separated out in room temperature, decompression suction filtration system and solid is washed with cold DMF 3 times, obtains white solid, 50
DEG C it is dried overnight, obtains white powdery solids (3- hydroxypropyls) tri-phenyl-phosphorus bromide；
(2) it is separately added into NH in the 100ml there-necked flasks equipped with magnetic agitation4HCO3(7.9g, 0.1mol), 40% formaldehyde
The aqueous solution (contain formaldehyde 0.1mol) and ethyl acetoacetate (26g, 0.2mol) and 60ml volumetric concentrations are 50% ethanol water,
Under nitrogen protection, 20h is heated to reflux, until there is yellowish precipitation, reaction system is cooled to room temperature, suction filtration solid and with cold
Ethanol is washed twice, and is dried to obtain compound 7 (wherein R is hydrogen) faint yellow solid (20.32g), is weighed 2.53g (0.01mol)
Compound 7 adds 100ml single port vials, is then separately added into 30ml methanol successively, 4mol/L NaOH aqueous solution 10ml, plus
Heat backflow 5h or so.Reaction solution is poured into 200ml cold water, is filtered under diminished pressure, filtrate adjusts pH=2.5, analysis with 1mol/L hydrochloric acid
Go out solid, be filtered under diminished pressure and be dried to obtain faint yellow solid compound 8 (wherein R is hydrogen) (1.52g).
(3) compound 8 (450mg, 2.0mmol), 1- hydroxy benzenes are added in the 100ml round-bottomed flasks equipped with magnetic agitation
And triazole (HOBT) (162mg, 1.2mmol) and DMF (15mL) stirring half an hours, add 1- ethyls-(3- dimethylaminos
Propyl group) phosphinylidyne diimmonium salt hydrochlorate (EDCI) (230mg, 1.2mmol), step 1) obtain (3- hydroxypropyls) tri-phenyl-phosphorus bromide
(514mg, 1mmol) and triethylamine (202mg, 2mmol).Lucifuge reaction under nitrogen protection is stayed overnight, and decompression steams most
DMF, by the NaHCO of saturation3The aqueous solution (20mL) is added in reaction system, separates out faint yellow solid, and decompression collected by suction is consolidated
Body, the method purified after drying by column chromatography obtains faint yellow solid, and (36.9%) 390mg, yield is.Solvent (oil
Ether：Ethyl acetate).
1H NMR(500MHz,DMSO-d6) δ=8.41 (s, 4H ,-OH), δ=7.91-7.70 (m, 15H ,-CH2-P+ Ph 3),
δ=7.39 (d, J=15.8Hz, 1H=CH- Ph), δ=7.11 (s, 1H, benzene), δ=6.90 (d, J=8.1Hz, 1H,
), benzene δ=6.73 (d, J=8.1Hz, 1H, benzene), δ=6.68 (s, 1H, benzene), δ=6.55 (d, J=
8.0Hz, 1H, benzene), δ=6.43 (d, J=8.1Hz, 1H, benzene), δ=6.22 (d, J=15.8Hz, 1H ,=CH-
), COO- δ=5.12 (dd, J=6.9,5.7Hz, 1H, COO-CH- COO), δ=4.16 (t, J=5.5Hz, 2H ,-COO-CH 2-
CH2), δ=3.54 (dd, J=6.3Hz, 2H, Ph-CH 2- CH-), δ=3.00-2.90 (m, 2H ,-P+-CH 2-), 1.81 (d, J=
7.3Hz,2H,-P+-CH2-CH 2-).13C NMR(500MHz,DMSO-d6):δ=18.365,22.157,36.783,64.520,
73.569,115.913,116.298,116.729,117.733,118.427,119.110,120.582,122.265,
125.408,127.048,130.882,131.065,134.171,134.251,135.712,135.735,145.234,
(146.107,146.920,147.370,150.445,166.875,170.102.ESI-MS m/z)=663.5 [M]+.
After nuclear magnetic resonance and mass spectral characteristi, obtain Mitochondrially targeted dihydrogen pyridine derivative crude product (shown in Formulas I, its
In：R is hydrogen atom, and X is bromine)
It is demonstrated experimentally that the 3- substituted with chloro- 1 propyl alcohol of equimolar 3- or iodo- 1 propyl alcohol of 3- in the present embodiment step (1) is bromo-
1 propyl alcohol, other same the present embodiment, the Mitochondrially targeted dihydrogen pyridine derivative crude product shown in preparation formula (I) is (wherein：R is that hydrogen is former
Son, X is chlorine or iodine).
Embodiment 2
A kind of preparation method of Mitochondrially targeted dihydrogen pyridine derivative (I-2), comprises the following steps：
(1) (1) of be the same as Example 1；
(2) with the formaldehyde in equimolar benzaldehyde alternate embodiment (2), the other the same as in Example 2；
(3) added in the 100ml round-bottomed flasks equipped with magnetic agitation compound 8 (wherein R be phenyl) (600mg,
2.0mmol), HOBT (162mg, 1.2mmol) and DMF (15mL) stirring half an hour, add EDCI (230mg,
1.2mmol), step 1) obtain (3- hydroxypropyls) tri-phenyl-phosphorus bromide (514mg, 1mmol) and triethylamine (202mg,
2mmol).Lucifuge reaction under nitrogen protection is stayed overnight, and decompression steams most DMF, by the NaHCO of saturation3The aqueous solution
(20mL) is added in reaction system, separates out faint yellow solid, depressurizes collected by suction solid, is purified after drying by column chromatography
Method obtains faint yellow solid (640mg, 52.9%).Solvent (petroleum ether：Ethyl acetate).
1H NMR(500MHz,DMSO-d6):δ=9.08 (s, 1H ,-NH), δ=7.98-7.57 (m, 15H ,-PPh 3), δ=
7.09 (d, J=7.2Hz, 2H, benzene), δ=6.99 (t, J=7.6Hz, 2H, benzene), δ=6.86 (t, J=
7.3Hz, 1H, benzene), δ=4.80 (s, 1H ,-CH-), δ=4.18-3.98 (m, 2H ,-CH 2-CH2-CH2-PPh3), δ=
3.97–3.89(m,2H,-CH2 CH3), δ=3.47-3.35 (m, 2H ,-CH2-CH2-CH 2-PPh3), δ=2.25 (s, 3H ,-
CH 3), δ=2.22 (s, 3H ,-CH 3), δ=1.75 (m, J=14.6,7.3Hz, 2H ,-CH2-CH 2-CH2-), δ=1.08 (t, J=
7.1Hz,3H,-CH2CH 3).13C NMR(500MHz,DMSO-d6):δ=14.09,18.08,18.19,38.74,58.92,
100.69,102.18,117.59,118.27,127.11,127.73,130.15,130.25,133.37,133.45,134.97,
(145.06,146.82,148.15,166.42,166.83.ESI-MS m/z)=604.6 [M]+
After nuclear magnetic resonance and mass spectral characteristi, obtain Mitochondrially targeted dihydrogen pyridine derivative crude product (shown in Formulas I, its
In：R is phenyl ring, and X is bromine)
It is demonstrated experimentally that the 3- substituted with chloro- 1 propyl alcohol of equimolar 3- or iodo- 1 propyl alcohol of 3- in the present embodiment step (1) is bromo-
1 propyl alcohol, other same the present embodiment, the Mitochondrially targeted dihydrogen pyridine derivative crude product shown in preparation formula (I) is (wherein：R is benzene
Ring, X is chlorine or iodine).
Embodiment 3
A kind of preparation method of Mitochondrially targeted dihydrogen pyridine derivative (I-3), comprises the following steps：
(1) (1) of be the same as Example 1；
(2) with the formaldehyde in equimolar thiophene 2- formaldehyde alternate embodiment (2), the other the same as in Example 2；
(3) added in the 100ml round-bottomed flasks equipped with magnetic agitation compound 8 (wherein R be thienyl) (307mg,
1.0mmol), HOBT (162mg, 1.2mmol) and DMF (15mL) stirring half an hour, add EDCI (230mg,
1.2mmol), step 1) obtain (3- hydroxypropyls) tri-phenyl-phosphorus bromide (514mg, 1mmol) and triethylamine (202mg,
2mmol).Lucifuge reaction under nitrogen protection is stayed overnight, and decompression steams most DMF, by the NaHCO of saturation3The aqueous solution
(20mL) is added in reaction system, separates out faint yellow solid, depressurizes collected by suction solid, is purified after drying by column chromatography
Method obtains faint yellow solid (470mg, 77%).Solvent (petroleum ether：Ethyl acetate).
1H NMR(500MHz,DMSO-d6):δ=9.33 (s, 1H ,-NH), 7.80 (dq, J=19.1,7.4Hz, 15H ,-
PPh 3), 7.04 (d, J=6.0Hz, 1H, thiophene), 6.68-6.61 (m, 1H, thiophene), 6.58 (d, J=
3.3Hz,1H,thiophene),5.13(s,1H,-CH-), 4.15 (dd, J=38.2,33.0Hz, 2H ,-CH 2-CH2-CH2-
PPh3),4.04–3.95(m,2H,-CH2 CH3), 3.40 (t, J=13.2Hz, 2H ,-CH2-CH2-CH 2-PPh3),2.26(s,
3H,-CH 3),2.23(s,3H,-CH 3),1.87–1.75(m,2H,-CH2-CH 2-CH2-), 1.12 (t, J=7.1Hz, 3H ,-
CH2CH 3).13C NMR(500MHz,DMSO-d6):δ=14.97,18.84,18.88,22.35,34.5,59.90,63.01,
63.16,101.00,102.40,118.40,119.09,123.13,124.12,127.07,130.96,131.06,134.17,
ESI-MS 134.25,135.74,135.76,146.50,148.07,152.70,166.97,167.36. (m/z)=610.5 [M
]+
After nuclear magnetic resonance and mass spectral characteristi, obtain Mitochondrially targeted dihydrogen pyridine derivative crude product (shown in Formulas I, its
In：R is thiphene ring, and X is bromine)
It is demonstrated experimentally that the 3- substituted with chloro- 1 propyl alcohol of equimolar 3- or iodo- 1 propyl alcohol of 3- in the present embodiment step (1) is bromo-
1 propyl alcohol, other same the present embodiment, the Mitochondrially targeted dihydrogen pyridine derivative crude product shown in preparation formula (I) is (wherein：R is thiophene
Ring, X is chlorine or iodine).
Embodiment 4
A kind of preparation method of Mitochondrially targeted dihydrogen pyridine derivative (II), comprises the following steps：
(1) (1) of be the same as Example 1；
(2) (3- hydroxypropyls) tri-phenyl-phosphorus bromide 2.0g is added in the 100ml there-necked flasks equipped with magnetic agitation
(5mmol), in hydrobromic acid (Han Liang≤40% is added dropwise at 108 DEG C) to just dissolving, back flow reaction 24 hours.It is cooled to room temperature,
Solution is slowly dropped in 50 times of v aqueous solutions, there is white precipitate precipitation.It is filtered under diminished pressure, cold water is washed, dries, obtain white solid
Body (3- bromopropyls) tri-phenyl-phosphorus bromide (1.9g, yield 83%).
(3) by (3- bromopropyls) tri-phenyl-phosphorus bromide 0.92g (20mmol) in 50ml round-bottomed flasks, 0.24g is added
The niacinamide (13) of (20mmol).In addition 10ml DMF at 100 DEG C.After backflow 5 hours, room temperature is cooled to.Vacuum distillation goes out
Most DMF solvent, adds suitable quantity of water (10ml) dissolving.Use ethyl acetate (40ml × 6), dichloromethane (40ml respectively again
× 6) extracting and washing, vacuum distillation goes out most of water, plus a small amount of methanol dissolving residue and is added to 50-100 times of volumes of acetic acid
In ethyl ester, there is white solid precipitation.Filtering, cold ethyl acetate is washed, and is dried, is obtained white solid 3- carbamyls -1- (3- (triphens
Base phosphine) propyl group) pyridiniujm (14) 0.76g.(yield 65%).
(4) 3- carbamyls -1- (3- (triphenylphosphine) propyl group) pyridiniujm (14) 0.58g (1mmol) is taken in round-bottomed flask
In, add 0.52g (3mmol) sodium dithionite.Dissolved with sodium carbonate liquor and adjust PH to 8-9 (after stable reaction
PH), lucifuge room temperature reaction 4h, depressurizes suction filtration solid, and with massive laundering, drying obtains faint yellow solid 0.36g (yields
70%).
1H NMR(500MHz DMSO-d6):δ=8.00-7.65 (m, 15H ,-+PPh 3),:δ=6.87 (s, 1H,
), pyridine δ=6.56 (s, 2H ,-NH 2), 5.84 (d, J=8.0Hz, 1H, pyridine), δ=4.61-4.54 (m, 1H,
), pyridine δ=3.57-3.44 (m, 2H ,-CH2-+PPh3), δ=3.26 (t, J=6.9Hz, 2H ,-CH2-CH2-CH 2-N),δ
=2.92 (d, J=1.0Hz, 2H, pyridine-CH 2-), δ=1.69 (dd, J=14.8,7.4Hz, 2H ,-CH2-CH 2-
CH2-) .ESI-MS (m/z)=427.8 [M]+
After nuclear magnetic resonance and mass spectral characteristi, the Mitochondrially targeted dihydrogen pyridine derivative crude product shown in formula (II) is obtained
(wherein：X is bromine)
It is demonstrated experimentally that being substituted with chloro- 1 propyl alcohol of equimolar 3- or 3-1 iodine propyl alcohol (11-3) in the present embodiment step (1)
Bromo- 1 propyl alcohol of 3-, other same the present embodiment, the Mitochondrially targeted dihydrogen pyridine derivative crude product shown in formula (II) is (wherein：X
For chlorine or iodine)
Experimental example 1
Dimethyl diaminophenazine chloride method determines cell survival rate
1st, cell culture
CHO-K1 cells (National Laboratory cell shared resource platform is bought) are 5%CO in volumetric concentration237 DEG C culture
Cultivated in case, the DMEM nutrient solutions culture containing 10% dual anti-hyclone to logarithmic phase.
2nd, cell survival rate is determined
It is layered on cultivating to the CHO-K1 cells of logarithmic phase with every 4000, hole in 96 orifice plates, culture is adherent overnight, irradiation
Preceding 30min is respectively to Mitochondrially targeted dihydro shown in Mitochondrially targeted dihydrogen pyridine derivative (I-1, I-2, I-3) and formula (II)
Pyridine derivate decoction (1 μm of ol/L, solvent is containing serum and dual anti-DMEM culture mediums) is placed in incubator per the μ L of hole 100
Middle culture 30min, is irradiated with 4Gy gamma-rays, is then continued to cultivate 24 hours in incubator, nutrient solution is removed, then per hole
The dimethyl diaminophenazine chloride nutrient solution for adding 100 μ L saturations cultivates 2h in incubator, cell is fully absorbed dimethyl diaminophenazine chloride, suctions out dimethyl diaminophenazine chloride training
Nutrient solution, is washed 2 times with 150 μ L phosphate buffers (PBS), washes away unabsorbed residual dimethyl diaminophenazine chloride, dries PBS, is added in 150 μ L
Red lysate (the acetic acid of property：Ethanol：Volume ratio=1 of water:50:49) 3min, is shaken, absorbance is determined in 544nm with ELIASA.
Experimental result：Mitochondrially targeted dihydrogen pyridine derivative can substantially increase the survival rate of cell after irradiation (see Fig. 1).
Experimental example 2
ROS elimination effects are determined
1. with the step 1 of experimental example 1；
2. will cultivate to the CHO-K1 cells of logarithmic phase, it is layered in 6 orifice plates, per hole 1x104It is individual, two groups of controls are set respectively
Group and four groups of dosing groups, dosing group in pre-irradiation 30min, be separately added into Mitochondrially targeted dihydrogen pyridine derivative (I-1, I-2,
I-3 (1 μm of ol/L, solvent is containing serum and dual anti-for Mitochondrially targeted dihydrogen pyridine derivative decoction) and shown in formula (II)
DMEM culture mediums) per hole 1mL, control group only adds the fresh of equivalent in pre-irradiation 30min and cultivated containing serum with dual anti-DMEM
Base, 30min is cultivated while being placed in incubator.Then irradiated, then cultivated 24 hours in incubator with 4Gy gamma-rays, removed
Nutrient solution, adds 2', 7'- dichlorofluorescins diethylester (DCFH-DA) (sigma is bought) (5 μM, culture medium dissolving) culture
Liquid 1ml, 37 DEG C of incubation 20min of lucifuge, removes DCFH-DA nutrient solutions, is washed with PBS 3 times, use collected by trypsinisation cell, enzyme
Mark instrument determines fluorescence intensity under the conditions of 488nm excitation wavelengths, 525nm launch wavelengths.
Experimental result：Mitochondrially targeted dihydrogen pyridine derivative Scavenging ability substantially increases (see Fig. 2).
Experimental example 3
DNA double chain fracture is determined
1. with the step 1 of experimental example 1；
2. will cultivate to the Chinese hamster ovary celI of logarithmic phase, one group is set not irradiate group and four groups of irradiation groups, four groups of irradiation groups respectively
Pre-irradiation 30min, respectively to Mitochondrially targeted shown in Mitochondrially targeted dihydrogen pyridine derivative (I-1, I-2, I-3) and formula (II)
Dihydrogen pyridine derivative decoction (1 μm of ol/L, solvent for containing serum and dual anti-DMEM culture mediums) and it is fresh containing serum and pair
Anti- DMEM culture mediums do not irradiate per the μ L of hole 500 and group add the fresh containing serum and dual anti-DMEM culture mediums of equivalent, four
Group irradiation group is irradiated with 4Gy gamma-rays respectively.Four groups of cells after irradiation and group is not irradiated culture 1 is small in incubator simultaneously
When, nutrient solution is removed, the paraformaldehyde aqueous solution of 500 μ L mass fractions 4%, fixed 15min, PBS washings 3 times, with 500 μ L is added
Mass fraction 0.2%, the solution of Triton-X 100, (solvent of the solution of Triton-X 100 is PBS) broken cell film 15min is used
PBS is washed 3 times, and the closing of sheep blood serum working solution 2h, 200 μ L rabbit polyclonal γ-H2AX (abcom is bought, and 1:1000 blood of goats are thin
Release) hatch 4 DEG C overnight, PBS is washed 3 times, 200 μ L goat-anti rabbit fluorescence secondary antibodies (abcom is bought, and 1:2000PBS dilutes), lucifuge room
Temperature is incubated 1h, and PBS is washed 3 times, DAPI (Suo Laibao is bought) dyeing 5min, PBS washing 3 times, and (Suo Laibao is purchased anti-quencher mounting
), taken pictures with AMG evo fluorescence microscopes, Image Pro Plus6.0 softwares focus is counted.
Experimental result：Mitochondrially targeted dihydrogen pyridine derivative protection DNA double chain ability increased.(see Fig. 3).
Experimental example 4
1. with the step 1 of experimental example 1；
2. by 106Individual cell is inoculated into laser co-focusing culture dish, after cell attachment, respectively to Mitochondrially targeted dihydro
(drug concentration is 10 to Mitochondrially targeted dihydrogen pyridine derivative decoction shown in pyridine derivate (I-1, I-2, I-3) and formula (II)- 5Mol/L) 37 DEG C of incubation 1h in incubator, remove cell culture fluid, and wash 3 times with PBS, and 37 DEG C of addition contains
Mitotracker Red@The cell culture fluid of (life companies buy) (20nmol/L), is incubated 20min under the conditions of 37 DEG C, discards
Former culture medium, adds PBS and washs 3 times, the paraformaldehyde for adding 4% is fixed, using in confocal laser scanning microscope cell
Medicine and Mitotracker Red@Distribution.
Experimental result：Tentatively conclude that Mitochondrially targeted dihydrogen pyridine derivative effectively can be gathered in mitochondria.(see
Fig. 4).
In our daily normal vital movements, body has Free-radical ring opening polymerization, including superoxide dismutase,
Catalase, glutathione reductase etc. removes the large biological molecule of free radical.But, on the one hand due to people itself aging
Cause the decline of Free-radical ring opening polymerization, the superoxide anion that mitochondrial oxidation breathing is produced, which can not be accessed, effectively to be removed,
Be diffused into cell and be converted to more active oxygens, cause vivo oxidation stress, so as to produce Parkinson's and alzheimer
The disease related to active oxygen such as disease；On the other hand due to being acted on by ionising radiation, mitochondrial oxidation respiratory chain and coding
The DNA of oxidation-respiration chain sustains damage, and will turn into the main place of active oxidative burst, if the active oxygen of excess generation is obtained not
Tool functional large biological molecule will be damaged to effectively removing, and then causes the Free-radical ring opening polymerization obstacle of human body, is made
Cellular oxidation stress.As can be seen here, the Mitochondrially targeted active oxygen radical of our exogenous intakes is removed under these conditions
These diseases related to active oxygen radical of medicine energy effectively preventing and dysfunction disease.And this research is due to mitochondria target
Scavenging capacity oxygen radical in mitochondria can be effectively gathered in dihydrogen pyridine derivative, therefore such compound can conduct
The curative of Parkinson's, alzheimer disease, while can also clinical as tumour radiotherapy in Nuclear Accident Emergency treatment
When radiation injury protection medicine.
Mitochondrially targeted dihydrogen pyridine derivative (I) or the inclusion compound of Mitochondrially targeted dihydrogen pyridine derivative (II), altogether
Brilliant, polycrystalline or solid dosage forms, liquid dosage form or semisolid dosage form containing said derivative belong to the protection model of the present invention
Enclose.
Solid dosage forms can be tablet (including ordinary tablet, dispersible tablet, effervescent tablet, chewable tablets etc.)；
Liquid dosage form can be solution (including true solution or colloidal solution), emulsion (including o/w types, w/o types and multiple
Breast), supensoid agent, injection (powder pin, liquid drugs injection)；
Semisolid dosage form can be paste etc..
Common system is made in Mitochondrially targeted dihydrogen pyridine derivative (I) or Mitochondrially targeted dihydrogen pyridine derivative (II)
Agent, sustained release preparation, controlled release preparation and various particulate delivery systems.
Claims (6)
1. Mitochondrially targeted dihydrogen pyridine derivative, it is characterized in that with following structure
Wherein：R is hydrogen atom, phenyl ring or thiophene；X is chlorine, bromine or iodine.
2. the preparation method of the Mitochondrially targeted dihydrogen pyridine derivative (I) of claim 1, it is characterized in that comprising the following steps：
1) by triphenylphosphine and 3- halogen propyl alcohol reaction generation (3- hydroxypropyls) triphenyl phosphonium halides；
2) by ammonium hydrogen carbonate, ethyl acetoacetate withThe back flow reaction in ethanol water, obtains compound (7), chemical combination
Thing (7) obtains compound (8) by sodium hydrate aqueous solution hydrolysis, compound (8) and step 1) (the 3- hydroxypropyls) three that obtains
Phenyl phosphorus Halides is anti-under 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt hydrochlorate and I-hydroxybenzotriazole catalysis
Should, obtain the dihydrogen pyridine derivative shown in formula (I)；
Reaction equation is：
Wherein：R is hydrogen atom, phenyl ring or thiophene；X is chlorine, bromine or iodine.
3. the preparation method of the Mitochondrially targeted dihydrogen pyridine derivative (II) of claim 1, it is characterized in that comprising the following steps：
By triphenylphosphine and 3- halogen propyl alcohol reaction generation (3- hydroxypropyls) triphenyl phosphonium halides；By (3- hydroxypropyls) triphenyl phosphonium halides with
HBr reactions obtain (3- bromopropyls) triphenyl phosphonium halides；By (3- bromopropyls) triphenyl phosphonium halides and niacinamide reactionization
Compound (14), under conditions of pH=8~9, by sodium hydrosulfite reducing compound (14), obtains the dihydropyridine shown in formula (II)
Derivative；
Reaction equation is：
Wherein：X is chlorine, bromine or iodine.
4. the Mitochondrially targeted dihydrogen pyridine derivative of claim 1 is preparing the application for the treatment of alzheimer medicine.
5. the Mitochondrially targeted dihydrogen pyridine derivative of claim 1 is preparing the application for the treatment of Parkinsonian drugs.
6. the Mitochondrially targeted dihydrogen pyridine derivative of claim 1 is preparing the application of radioprotector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710436442.4A CN107216352B (en) | 2017-06-11 | 2017-06-11 | Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710436442.4A CN107216352B (en) | 2017-06-11 | 2017-06-11 | Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107216352A true CN107216352A (en) | 2017-09-29 |
| CN107216352B CN107216352B (en) | 2019-05-17 |
Family
ID=59947402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710436442.4A Active CN107216352B (en) | 2017-06-11 | 2017-06-11 | Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107216352B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108969504A (en) * | 2018-06-22 | 2018-12-11 | 中国人民解放军第四军医大学 | The preparation method of Mitochondrially targeted nm radiation protection medicine |
| CN108997422A (en) * | 2018-06-22 | 2018-12-14 | 中国人民解放军第四军医大学 | The preparation of Mitochondrially targeted radioprotectant and its application in radiation injury protection |
| CN111759861A (en) * | 2019-03-27 | 2020-10-13 | 台湾粒线体应用技术股份有限公司 | Use of mitochondria for the preparation of a pharmaceutical composition for the treatment and/or prevention of alzheimer's disease |
| US20210395280A1 (en) * | 2018-11-01 | 2021-12-23 | The Medical College Of Wisconsin, Inc. | Targeting Redox-Active Pyridinium Cations to Mitochondria to Inhibit Proliferation of Drug-Resistant Cancer Cells |
| WO2022114187A1 (en) * | 2020-11-30 | 2022-06-02 | 国立大学法人九州大学 | Compound having inhibitory action against excessive mitochondrial divisions |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100099741A1 (en) * | 2008-10-20 | 2010-04-22 | Effat Emamian | Molecular Targets for Treatment of Learning and Memory Dysfunction |
| CN105693769A (en) * | 2016-03-11 | 2016-06-22 | 中国医学科学院放射医学研究所 | Rosmarinic acid derivative, preparation method and application |
-
2017
- 2017-06-11 CN CN201710436442.4A patent/CN107216352B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100099741A1 (en) * | 2008-10-20 | 2010-04-22 | Effat Emamian | Molecular Targets for Treatment of Learning and Memory Dysfunction |
| CN105693769A (en) * | 2016-03-11 | 2016-06-22 | 中国医学科学院放射医学研究所 | Rosmarinic acid derivative, preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| ANDREY S.TSVETKOV等: "A small-molecule scaffold induces autophagy in primary neurons and protects against toxicity in a Huntington disease model", 《PNAS》 * |
| 张宇睿等: "1，4-二氢吡啶类化合物对CHO-K1细胞的辐射防护作用", 《辐射研究与辐射工艺学报》 * |
| 王华伟 等: "线粒体靶向药物的研究进展", 《国际放射医学核医学杂志》 * |
| 路军 等: "1，4-二氢吡啶衍生物合成方法的改进和芳构化研究", 《有机化学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108969504A (en) * | 2018-06-22 | 2018-12-11 | 中国人民解放军第四军医大学 | The preparation method of Mitochondrially targeted nm radiation protection medicine |
| CN108997422A (en) * | 2018-06-22 | 2018-12-14 | 中国人民解放军第四军医大学 | The preparation of Mitochondrially targeted radioprotectant and its application in radiation injury protection |
| US20210395280A1 (en) * | 2018-11-01 | 2021-12-23 | The Medical College Of Wisconsin, Inc. | Targeting Redox-Active Pyridinium Cations to Mitochondria to Inhibit Proliferation of Drug-Resistant Cancer Cells |
| CN111759861A (en) * | 2019-03-27 | 2020-10-13 | 台湾粒线体应用技术股份有限公司 | Use of mitochondria for the preparation of a pharmaceutical composition for the treatment and/or prevention of alzheimer's disease |
| CN111759861B (en) * | 2019-03-27 | 2023-01-13 | 台湾粒线体应用技术股份有限公司 | Use of mitochondria for the preparation of a pharmaceutical composition for the treatment and/or prevention of alzheimer's disease |
| WO2022114187A1 (en) * | 2020-11-30 | 2022-06-02 | 国立大学法人九州大学 | Compound having inhibitory action against excessive mitochondrial divisions |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107216352B (en) | 2019-05-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107216352B (en) | Mitochondrially targeted dihydrogen pyridine derivative and preparation method and application | |
| CN100596299C (en) | Dehydrosilibinin diester derivatives, preparation method and use thereof | |
| CN100534992C (en) | Silybin esters derivatives and preparation and use thereof | |
| CN101255119B (en) | Novel tetrahydro curcumin derivatives and salt | |
| CN104387412B (en) | Fluorine boron two pyrrole derivative that erlotinib is modified and Synthesis and applications thereof | |
| CN110642852B (en) | Organic AIE photosensitive probe with mitochondrion targeting function and preparation method and application thereof | |
| CN111303139B (en) | Compound with aggregation-induced emission performance and preparation method and application thereof | |
| CN104447406B (en) | Alpha-cyano-4-hydroxy cinnamic acid derivative and preparation method and application thereof | |
| CN113956242B (en) | Photosensitizer for inducing immunogenic cell death and preparation method and application thereof | |
| CN111592482B (en) | PH reversible activation type photo-thermal/photodynamic/fluorescent integrated probe molecule | |
| CN105308040B (en) | 1,3- diamino cyclopentane formamide derivative | |
| CN109796483A (en) | A kind of water-soluble cationic photosensitizer and its preparation and application | |
| CN105693769B (en) | Rosmarinic acid derivative and preparation method and application | |
| CN113683602B (en) | Heptamethine cyanine micromolecule for multi-modal treatment of hypoxic tumors, and preparation method and application thereof | |
| CN103360456B (en) | Triterpene compound and Synthesis and applications | |
| CN106565763A (en) | pH sensitive axially substituted silicon phthalocyanine complex, preparing method of pH sensitive axially substituted silicon phthalocyanine complex and application of pH sensitive axially substituted silicon phthalocyanine complex to medicines | |
| CN106892920A (en) | Aloperine derivative, Preparation Method And The Use | |
| CN102203088A (en) | Poly-heteroaryl derivatives for the treatment of cancer | |
| CN104151339B (en) | Boron-containing acridine derivative as well as preparation method and application thereof | |
| CN102633796B (en) | New preparation method of sophora flavescens acid derivative | |
| CN114656453A (en) | Heptamethine indole cyanine-TEMPO chemical couple chain small molecule, preparation method and application thereof in preparing radioprotection preparation | |
| CN102702297B (en) | Preparation method of cholic acid-naphthalimide compound | |
| CN103467452B (en) | A kind of benzamide compound and application thereof | |
| CN105541732A (en) | Beta-lapachone derivative, and preparation method and medicinal application thereof | |
| CN102241665B (en) | 4-(4,5-dimethoxycarboxyl-1,3-dithiolane-2-yl)benzoyl-L-amino acid benzyl ester as well as synthesis method and applications thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |