CN105218377A - A kind of Hydroxytyrosol NO donor derivative and its preparation method and application - Google Patents
A kind of Hydroxytyrosol NO donor derivative and its preparation method and application Download PDFInfo
- Publication number
- CN105218377A CN105218377A CN201510582108.0A CN201510582108A CN105218377A CN 105218377 A CN105218377 A CN 105218377A CN 201510582108 A CN201510582108 A CN 201510582108A CN 105218377 A CN105218377 A CN 105218377A
- Authority
- CN
- China
- Prior art keywords
- dioxole
- dihydroxy
- benzene
- phenylbenzene
- benzos
- 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
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of Hydroxytyrosol NO donor derivative and its preparation method and application, it is for the preparation of the medicine in hypoglycemic and blood vessel endothelium protection.Confirm through inside and outside pharmacological evaluation, above-claimed cpd has following pharmacologically active: (1) scavenging capacity oxyradical; (2) NO is discharged, vasodilator; (3) suppress glucosidase activity, have certain hypoglycemic activity.Internal pharmacokinetics experimental result shows that it can discharge NO rapidly in vivo, makes it reach higher level.The invention provides the preparation method of Hydroxytyrosol NO donor derivative, can obtain changing derivative, have raw material and be easy to get by 4 step organic syntheses, reaction conditions is gentle, and reaction process is simple to operate, the advantage that agents useful for same is cheap.This derivative has obvious hypoglycemic and vasorelaxation action, can be used for preventing and treating diabetes and vascular complication thereof, has larger clinical value.
Description
Technical field:
The invention belongs to new drug development field, be specifically related to a kind of Hydroxytyrosol NO donor derivative and preparation method thereof and the application in hypoglycemic and blood vessel endothelium protection.
Background technology:
Hydroxytyrosol (hydroxytyrosol, HT) chemical name 3,4-dihydroxyphenyl ethanol, it is a kind of natural polyphenol compounds in sweet oil with strong anti-oxidation, in the fruit being extensively present in Oleaceae plants olive and branches and leaves, to various active oxygen, there is eliminating effect, there is stronger anti-oxidant activity.In addition, large quantity research confirms that Hydroxytyrosol also has good effect in prevention cardiovascular and cerebrovascular diseases, anti-inflammatory are anticancer etc.Hydroxytyrosol, because of its special biology and pharmacologically active, has been widely used in foodstuffs industry, nutritive health-care and makeup.The capsule of Hydroxytyrosol and tablet has in succession been had to come out at present abroad.
Nitrogen protoxide (NO) donator type medicine is in vivo through enzyme or non-enzymatic effect release NO.Classical medicine is that the single structure such as pannonit, Sodium Nitroprusside directly discharges NO, forms medicine at present increasingly receive publicity by various linking group and known drug or active pharmacophore and NO donor.Prove: give NO substrate or exogenous NO donor medicine can obviously improve Vasodilation in vascular lesion and delay pathological process.What Merck company of the U.S. was studied enters II phase clinical NO-donator type cox 2 inhibitor, is exactly to protect vascular endothelial cell by release NO, but also can reduce gastrointestinal tract mucous stimulation.The NO donor medicine acting on NO link at present improves one of endothelial function effective ways.
Summary of the invention:
The object of the present invention is to provide a kind of Hydroxytyrosol NO donor derivative and preparation method thereof, and it is reducing the application in blood sugar, anti-oxidation stress and reparation endothelial function disturbance.
For achieving the above object, the present invention adopts following technical scheme to realize:
A kind of Hydroxytyrosol NO donor derivative, its chemical structural formula is:
R is
wherein n=1 ~ 6.
The preparation method of above-mentioned a kind of Hydroxytyrosol NO donor derivative, comprising:
As n=1, synthesis step is as follows:
1) 3,4-dihydroxybenzyl alcohols are substituted obtained 4-chloromethyl-1, the 2-dihydroxy-benzene of reaction;
2) 4-chloromethyl-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole;
3) 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole;
4) 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole obtains 4-nitrooxymethyl through acidic hydrolysis)-1,2-dihydroxy-benzene;
As n=2, synthesis step is as follows:
1) Hydroxytyrosol is substituted obtained 4-(2-chloroethyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(2-chloroethyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=3, synthesis step is as follows:
1) 3,4-dihydroxy-benzene propyl alcohol are substituted obtained 4-(3-chloropropyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(3-chloropropyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(3-nitre acryloxypropylethoxysilane)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=4, synthesis step is as follows:
1) 3,4-dihydroxy-benzene butanols are substituted obtained 4-(4-chlorobutyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(4-chlorobutyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(4-nitre acyloxy butyl)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=5, synthesis step is as follows:
1) 3,4-dihydroxy-benzene amylalcohols are substituted obtained 4-(5-chlorine amyl group)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(5-chlorine amyl group)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
3) 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(5-nitre acyloxy amyl group)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=6, synthesis step is as follows:
1) obtained 4-(6-chlorine hexyl)-1, the 2-dihydroxy-benzene of reaction is substituted;
2) 4-(6-chlorine hexyl)-1,3,4-dihydroxy-benzene hexanol 2-dihydroxy-benzene and dichloro diphenyl methane react obtained 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
3) 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(6-nitre acyloxy hexyl)-1,2-dihydroxy-benzene through acidic hydrolysis.
The application of above-mentioned a kind of Hydroxytyrosol NO donor derivative, the application in the medicine preparing prevention and therapy diabetic vascular complications.
The present invention further improves and is: as the application of antioxidant.
The present invention further improves and is: the application in preparation vasodilation medicine.
The present invention further improves and is: protecting the application in medicine for the preparation of blood vessel endothelium.
The present invention further improves and is: preparing the application in hypoglycemic drug.
Relative to prior art, the present invention has following advantage:
The present invention is with natural polyphenol compounds Hydroxytyrosol for parent nucleus, and its 4 side chains are introduced the carbochain of different lengths, synthesizes a series of Hydroxytyrosol NO donor derivative.By 4 step organic syntheses, have raw material and be easy to get, reaction conditions is gentle, and reaction process is simple to operate, the advantage that agents useful for same is cheap.The present invention confirms through inside and outside pharmacological evaluation, and above-claimed cpd has following pharmacologically active: (1) scavenging capacity oxyradical; (2) NO is discharged, vasodilator; (3) suppress glucosidase activity, have certain hypoglycemic activity.Internal pharmacokinetics experimental result shows that it can discharge NO rapidly in vivo, makes it reach higher level.
Hydroxytyrosol NO donor derivative provided by the invention has good hypoglycemic and vasorelaxation action, can be used for preventing and treating diabetes and vascular complication thereof, has larger clinical value.
Accompanying drawing illustrates:
Fig. 1 is the synthetic route chart of 4-(2-nitrooxymethyl)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 2 is the synthetic route chart of 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 3 is the synthetic route chart of 4-(2-nitre acryloxypropylethoxysilane)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 4 is the synthetic route chart of 4-(2-nitre acyloxy butyl)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 5 is the synthetic route chart of 4-(2-nitre acyloxy amyl group)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 6 is the synthetic route chart of 4-(2-nitre acyloxy hexyl)-1,2-dihydroxy-benzene;
What mark in figure is specially:
(a)CCl
4，PPh
3，MeCN；(b)Ph
2CCl
2，PhMe；(c)AgNO
3，MeCN；(d)AcOH：H
2O(4:1)。
Fig. 7 is the external NO release profiles of HT-NO.
Fig. 8 is the impact of HT-NO on rat chest aorta ring diastole activity.
Fig. 9 is the impact of HT-NO on Human umbilical vein endothelial cells activity.
Figure 10 is the impact of HT-NO on the Human umbilical vein endothelial cells activity of high sugar induction.
Figure 11 is the impact of HT-NO on Human umbilical vein endothelial cells NO level.
Figure 12 is the impact of HT-NO on the Human umbilical vein endothelial cells NO level of high sugar induction.
Figure 13-1 is that HT adds the 1% xitix concentration curve of frozen a week under-20 DEG C of conditions.
Figure 13-2 is that HT-NO adds the 1% xitix concentration curve of frozen a week under-20 DEG C of conditions.
Figure 14 is the blood concentration-time curve of rat oral gavage HT.
Figure 15 is the blood concentration-time curve of rat oral gavage HT-NO.
Figure 16 is the blood concentration-time curve of HT after rat oral gavage HT-NO.
Figure 17 is the horizontal change curve of NO in blood plasma in rat oral gavage HT-NO, HT and physiological saline 24h.
Embodiment:
The invention will be further described to use embodiment below, it should be noted that these embodiments only for being illustrated the present invention, should not be construed as limitation of the present invention.
The synthesis step of HT-NO:
Embodiment 1
The synthesis of 4-chloromethyl-1,2-dihydroxy-benzene
Be dissolved in by 3,4-dihydroxybenzyl alcohol in 25mL anhydrous acetonitrile, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-chloromethyl-1,2-dihydroxy-benzene;
The synthesis of 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole
4-chloromethyl-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole;
The synthesis of 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole
By 4-chloromethyl-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole;
The synthesis of 4-nitrooxymethyl-1,2-dihydroxy-benzene
4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, after question response is complete, is cooled to room temperature, adds ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-nitrooxymethyl-1,2-dihydroxy-benzene.
Embodiment 2
The synthesis of 4-(2-chloroethyl)-1,2-dihydroxy-benzene
Be dissolved in by Hydroxytyrosol in 25mL anhydrous acetonitrile, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(2-chloroethyl)-1,2-dihydroxy-benzene;
The synthesis of 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole
4-(2-chloroethyl)-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole;
The synthesis of 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole
By 5-(2-chloroethyl)-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene:
5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, but after reacting completely, be cooled to room temperature, add ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene.
Embodiment 3
The synthesis of 4-(3-chloropropyl)-1,2-dihydroxy-benzene
Be dissolved in 25mL anhydrous acetonitrile by 3,4-dihydroxy-benzene propyl alcohol, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(3-chloropropyl)-1,2-dihydroxy-benzene;
The synthesis of 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole
4-(3-chloropropyl)-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole;
The synthesis of 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole
By 6-(3-chloropropyl)-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 4-(3-nitre acryloxypropylethoxysilane)-1,2-dihydroxy-benzene:
6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, but after reacting completely, be cooled to room temperature, add ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(3-nitre acryloxypropylethoxysilane)-1,2-dihydroxy-benzene.
Embodiment 4
The synthesis of 4-(4-chlorobutyl)-1,2-dihydroxy-benzene
Be dissolved in 25mL anhydrous acetonitrile by 3,4-dihydroxy-benzene butanols, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(4-chlorobutyl)-1,2-dihydroxy-benzene;
The synthesis of 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole
4-(4-chlorobutyl)-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole;
The synthesis of 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole
By 7-(4-chlorobutyl)-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 4-(4-nitre acyloxy butyl)-1,2-dihydroxy-benzene:
7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, but after reacting completely, be cooled to room temperature, add ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(4-nitre acyloxy butyl)-1,2-dihydroxy-benzene.
Embodiment 5
The synthesis of 4-(5-chlorine amyl group)-1,2-dihydroxy-benzene
Be dissolved in 25mL anhydrous acetonitrile by 3,4-dihydroxy-benzene amylalcohol, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(5-chlorine amyl group)-1,2-dihydroxy-benzene;
The synthesis of 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole
4-(5-chlorine amyl group)-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole
By 8-(5-chlorine amyl group)-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 4-(5-nitre acyloxy amyl group)-1,2-dihydroxy-benzene:
8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, but after reacting completely, be cooled to room temperature, add ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(5-nitre acyloxy amyl group)-1,2-dihydroxy-benzene.
Embodiment 6
The synthesis of 4-(6-chlorine hexyl)-1,2-dihydroxy-benzene
Be dissolved in 25mL anhydrous acetonitrile by 3,4-dihydroxy-benzene hexanol, add triphenyl phosphorus, tetracol phenixin successively, nitrogen protection, stirring at room temperature 10h, after question response is complete, decompression removing acetonitrile solvent, adds H
2o (20mL), with dichloromethane extraction, saturated NaCl solution washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(6-chlorine hexyl)-1,2-dihydroxy-benzene;
The synthesis of 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole
4-(6-chlorine hexyl)-1,2-dihydroxy-benzene is dissolved in toluene, adds dichloro diphenyl methane, nitrogen protection, return stirring 24h, after question response is complete, be cooled to room temperature, reaction system H
2o (50mL) washs, and aqueous phase toluene (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole
By 9-(6-chlorine hexyl)-2; 2-phenylbenzene benzo [1; 3] dioxole is dissolved in 25mL anhydrous acetonitrile, adds Silver Nitrate (2.50g), under nitrogen protection; lucifuge; be stirred at 57 DEG C and react completely, add NaCl solution stirring at room temperature 1h, filter away solid; decompression removing acetonitrile solvent, adds H
2o (20mL), with ethyl acetate (20mL × 3) extraction, merge organic phase, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
The synthesis of 4-(6-nitre acyloxy hexyl)-1,2-dihydroxy-benzene:
9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole is joined AcOH/H
2in the mixing solutions of O (20mL/5mL), under nitrogen protection, return stirring 4-5h, but after reacting completely, be cooled to room temperature, add ethyl acetate (50mL) and H
2o (50mL), organic phase saturated solution of sodium bicarbonate (20mL × 3) washing, anhydrous sodium sulfate drying, filter, concentrating under reduced pressure obtains yellow oil.Column chromatography for separation obtains 4-(6-nitre acyloxy hexyl)-1,2-dihydroxy-benzene.
Embodiment 7
HT-NO anti-oxidant activity is tested
Adopt scavenging free radicals method ([DPPH] method), the anti-oxidant activity of 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene (hereinafter abbreviated as HT-NO) is tested through row.
1 materials and methods
1.1 Experimental agents
Embodiment 2 prepares gained compound
Experiment material:
Reagent:
1,1-phenylbenzene-2-trinitrophenyl-hydrazine (1,1-Diphenyl-2-picrylhydrazyl [DPPH] (the upper graceful bio tech ltd of Hypon); Dehydrated alcohol (Tianjin Tian Li chemical reagent company limited);
Key instrument: 752 type ultraviolet-visible pectrophotometers (Shanghai Spectrum Apparatus Co., Ltd.); WH-2 micro-whirlpool mixed instrument (Shanghai Hu Xi analytical instrument Co., Ltd., Factory);
1.2 experimental technique
The preparation of detected sample:
1) [DPPH] and sample sets (Ai): adding 2mL concentration in the tool edge test tube of 10mL is 0.08mg/mL [DPPH] ethanolic soln, adds testing sample (HT-NO) and the positive control (0.1mg/mL xitix) of 2mL different concns respectively;
2) [DPPH] and group of solvents (Ao): adding 2mL concentration in the tool edge test tube of 10mL is 0.08mg/mL [DPPH] ethanolic soln and 2mL dehydrated alcohol;
3) sample and group of solvents (Aj): add 2mL dehydrated alcohol in the tool edge test tube of 10mL, adds testing sample (HT-NO) and the positive control (0.1mg/mL xitix) of 2mL different concns respectively;
After detected sample application of sample, shake up, room temperature leaves standstill 30min, detects light absorption value when 517nm wavelength.Calculate free radical scavenging activity according to the following formula.
[DPPH] free radical scavenging activity S%=[1-(Ai-Aj)/Ao] × 100%.
1.3 experimental result
Use SPSS matched curve and calculate IC
50, obtain HT-NO to [DPPH] half elimination ratio IC
50be 3.28 × 10
-5molL
-1, xitix is to [DPPH] half elimination ratio IC
50be 6.95 × 10
-5molL
-1.Result shows, and HT-NO has good radical scavenging activity.Result as table 1, table 2
Table 1 [DPPH] method measures the anti-oxidant activity result (%) of HT-NO
Table 2 [DPPH] method measures the anti-oxidant activity result (%) of xitix
\
Embodiment 8
The external NO of HT-NO discharges activity experiment
Adopt the external NO release of Griess method to HT-NO active in row test.
1 materials and methods
1.1 Experimental agents
Embodiment 2 prepares gained compound
Experiment material:
Reagent: anhydrous Sulphanilic Acid is received (Tianjin good fortune chemical reagent factory in morning); N-1-naphthodiamide hydrochloride (Tianjin Aoran Fine Chemical Research Institute); Sodium Nitrite (Tianjin good fortune chemical reagent factory in morning); Cys (Tianjin good fortune chemical reagent factory in morning); Potassium primary phosphate (Guangdong Guanghua Science and Technology Co., Ltd.); Sodium hydroxide (Hedong District, Tianjin red rock chemical reagent work); Phosphoric acid (Xi'an chemical reagent factory).
Key instrument: 752 type ultraviolet-visible pectrophotometers (Shanghai Spectrum Apparatus Co., Ltd.'s manufacture); WH-2 micro-whirlpool mixed instrument (manufacture of Shanghai Hu Xi analytical instrument Co., Ltd., Factory);
1.2 experimental technique
(1) drafting of NO release standard curve: phosphate buffer soln 2.5mL, methyl alcohol 2.5mL, equal-volume mixes, then adds the NaNO of different concns
2 -cys solution with the 0.1mol/L of 50 μ L, adds Griess reagent after mixing, mixing leaves standstill 10min, measures its absorbancy under 540nm.Use SPSS fit standard curve, and obtain typical curve equation by fitting of a curve.
(2) the external NO release of HT-NO measures: phosphate buffer soln 2.5mL, methyl alcohol 2.5mL, equal-volume mixes, at the Cys solution of the 0.1mol/L of the HT-NO and 50 μ L that add different concns, Griess reagent is added after mixing, mixing leaves standstill 10min, measures its absorbancy under 540nm.Use SPSS matched curve, and obtain corresponding NO by typical curve equation
2 -concentration, in Sodium Nitroprusside and hydroxytyrosol derivatives release NO efficiency=NO release concentration and initial reaction liquid, 0.1mM derivative contains the ratio of the mole number of NO.
1.3 experimental result
Result display HT-NO can discharge certain density NO in vitro.Result is as Fig. 7
Embodiment 9
HT-NO tests the impact of Endothelial Function
1 materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Sodium Nitroprusside, SD rat, computer bio signal record analysis system, tonotransducer, in vitro tissue organ constant temperature perfusion system.
1.2 experimental technique
Cervical dislocation puts to death rat, takes out aorta rapidly, is placed in the 4 DEG C of tyrode's solutions being connected with mixed gas, rejects fat and the reticular tissue of vessel outer wall.Vascular strip is cut into segment, every section of about 2 ~ 3mm; Get one section of vascular strip, through fixing spillikin, then hung up by blood vessel with little hook, the thin cotton thread of little hook is connected to sensor.Finally blood vessel is soaked in and tyrode's solution is housed, be connected with mixed gas and keep in the reaction tank of 37 DEG C of constant temperature.By software, the state of tension of monitoring vascular strip, is adjusted to about 0.8g by the pulling force of equilibrium state; Balance 1h, every 15min change tyrode's solution 1 time.3 times are stimulated with the KCl of 60mmol, when vasoconstriction arrives the most by force and stablizes, add the HT-NO of different concns, with physiological saline in contrast, the ratio between the maximum shrinkage amplitude of the vascular circle brought out with the antiotasis amplitude after adding medicine 15min and KCl reflects the change of antiotasis.
1.3 experimental result
The contraction that result display HT-NO can suppress KCl to cause vascular circle.Result is as Fig. 8
Embodiment 10
HT-NO is on the impact experiment of the Human umbilical vein endothelial cells activity of high sugar induction
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
DMEM culture medium dry powder (Gibco, USA), foetal calf serum (FBS) (Gibco, USA), pancreatin (Trypsin) (Beyotime), MTT (Sigma), glucose (Sigma), human umbilical vein endothelial (HUVECs), microplate reader (ThermoScientific)
1.2 experimental technique
1.2.1 cell cultures: cultivate HUVECs cell with DMEM (Dulbecco ' sModifiedEagle ' sMedium) nutrient solution containing 10%FBS, every 1-2 days goes down to posterity 1 time.
1.2.2 HUVECs cell is inoculated in 96 orifice plates in units of 5000-6000/hole, is divided into blank group, high sugar group and administration group.Model group gives the glucose solution of 55mM, and the height sugar of HT-NO (12.5,25,50,100,200 μMs) and 55mM that administration group gives different concns hatches 48h jointly.After hatching end, every hole adds 20 μ LMTT, hatches 4h, under microplate reader 492nm wavelength measures absorbancy (A) after dissolving with DMSO.Cytoactive (%) is calculated: cytoactive (%)=(the blank group of A experimental group-A)/(the blank group of A control group-A) × 100%, calculates corresponding cytoactive by following method.
1.3 experimental result
Result display gives separately HT-NO effect endotheliocyte, can promote cell proliferation, as Fig. 9 within the scope of finite concentration; And after giving the high sugar stimulation of endotheliocyte, cytoactive obviously reduces, give HT-NO and can play certain provide protection, as Figure 10 to the cell injury of high sugar induction.
Embodiment 11
HT-NO is on the impact experiment of the Human umbilical vein endothelial cells NO level of high sugar induction
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
DMEM culture medium dry powder (Gibco, USA), foetal calf serum (FBS) (Gibco, USA), pancreatin (Trypsin) (Beyotime), MTT (Sigma), glucose (Sigma), human umbilical vein endothelial (HUVECs), microplate reader (ThermoScientific)
1.2 experimental technique
1.2.1 cell cultures: cultivate HUVECs cell with DMEM (Dulbecco ' sModifiedEagle ' sMedium) nutrient solution containing 10%FBS, every 1-2 days goes down to posterity 1 time.
1.2.2 HUVECs cell is inoculated in 96 orifice plates in units of 5000-6000/hole, is divided into blank group, high sugar group and administration group.Model group gives the glucose solution of 55mM, and the height sugar of HT-NO (12.5,25,50,100,200 μMs) and 55mM that administration group gives different concns hatches 48h jointly.After hatching end, collecting cell supernatant liquor, measures absorbancy (A) by NO detection kit under microplate reader 550nm wavelength.Corresponding NO concentration is calculated by detection kit specification sheets.
1.3 experimental result
Result display gives separately HT-NO effect endotheliocyte, can promote the release of NO in cell conditioned medium liquid, as Figure 11 within the scope of finite concentration; And after giving the high sugar stimulation of endotheliocyte, NO content reduces in cell conditioned medium liquid, give the level that HT-NO can raise NO in the cell conditioned medium liquid of high sugar induction, as Figure 12.
Embodiment 12
The external glucoside inhibiting activity experiment of HT-NO
1 materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Acarbose (Bayer A.G, 50mg/ sheet, commercially available), glucose determination reagent box (Bioengineering Research Institute is built up in Nanjing), alpha-glucosidase (Sigma), 4-oil of mirbane-α-D-glucopyranoside (α-PNPG) (Sigma), 752 type ultraviolet-visible pectrophotometers (Shanghai Spectrum Apparatus Co., Ltd.'s manufacture)
1.2 experimental technique
1.2.1 suppress yeast α-glucosidase
(1) α-PNPG solution preparation is prepared; Alpha-glucosaccharase enzyme solution 10U/mL; NaCO3 solution 1mol/L; The target compound liquid to be measured (dissolving with the DMSO-phosphate buffered saline buffer that volume ratio is 1:9) of different concns.
(2) get alpha-glucosaccharase enzyme solution 10 μ L, in 37 DEG C of water-baths, hatch 10min, add the solution to be measured 50 μ L of different concns respectively, mix follow-up continuation of insurance temperature 10min, add α-PNPG solution 20 μ L again, after mixing, be incubated 15min, add NaCO3 solution 50 μ L termination reaction.Set up blank (not containing enzyme and sample) and negative control group (not containing sample), acarbose is positive control.Solution absorbance is detected at 405nm place, inhibiting rate (%) is calculated: inhibiting rate (%)=(A negative control-A sample)/(A negative control-A blank) × 100%, calculates corresponding half-inhibition concentration (IC by following method
50).
1.2.2 small intestine in rats alpha-glycosidase is suppressed
(1) preparation of small intestine in rats enzyme liquid: Rat Fast is de-cervical vertebra execution after can't help water 12h, and take out small intestine, rinse enteric cavity with cold saline, be placed in by clean small intestine on ice platform, scraping mucous membrane of small intestine, weighs.Add the phosphate buffered saline buffer of 4 DEG C of 5 times amount, homogenate in ice bath, then the centrifugal 20min of 5000r/min, gets supernatant.
(2) get enzyme liquid 10 μ L, hatch 15min in 37 DEG C of water-baths after, add the inhibitor 50 μ L of different concns, continue insulation 30min, add the maltose 20 μ L of 50mmol/L again, react 60min in 37 DEG C of water-baths after, take out and be placed in boiling water bath termination reaction; Do blank (without inhibitor), acarbose is positive control simultaneously.Extract reaction solution 10 μ in EP pipe, then add A test solution 500 μ L, B test solution 500 μ L in Reagent kit of glucose and mix, after 37 DEG C of insulation 15min, under 505nm, measure its absorbancy.Inhibiting rate (%) is calculated: inhibiting rate (%)=(A blank-A sample)/A blank × 100%, calculates corresponding half-inhibition concentration (IC by following method
50).
1.3 experimental result
The compound of result display preparation can suppress yeast α-glucosidase and small intestine in rats alpha-glycosidase.Result is as table 3
Table 3HT-NO is to the restraining effect IC of alpha-glycosidase
50(μM)
Embodiment 13
HT-NO tests the impact of the blood glucose in diabetic mice that STZ induces
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Reagent: STZ (Sigma), N1,N1-Dimethylbiguanide bulk drug (Tokyo HuaCheng Industry Co., Ltd), Hydroxytyrosol (Shaanxi Ying Hua Bioisystech Co., Ltd), the stable blood glucose meter of three promises and blood sugar test paper bar (Sinocare Biosensing Co., Ltd)
Laboratory animal: kunming mice, weight 25-30g, is provided by Xi'an Communications University's animal experimental center.
1.2 experimental technique
1.2.1 diabetic mouse model is set up
After 50 the mouse adaptability bought are fed 3 days, therefrom randomly draw 8 as Normal group, give chow diet to feed, all the other 42 overnight fastings, then give a certain amount of STZ citric acid-sodium citrate of experiment mice abdominal cavity injection location (160mgkg
-1).The citric acid-sodium citrate buffer of same dose volume also gives the injected in mice of Normal group.The equal fasting of all mouse in the 5th day after injection, the mouse tail vein of modeling is got blood and is measured its fasting plasma glucose, with mouse fasting blood sugar>=16.7mmolL
-1for the successful standard of diabetic experimental mouse model.
1.2.2 laboratory animal grouping and administration
Successful for modeling mouse is divided at random: model group, N1,N1-Dimethylbiguanide group (250mgkg
-1), hydroxytyrosol group (77mgkg
-1) and HT-NO group (100mgkg
-1), often organize mouse 8.Continuous gavage 4 weeks, normal group and model group give the physiological saline gavage of equal volume.Observe the visual condition of mouse every day, record each group of amount of drinking water and dietary amount simultaneously.
1.3 experimental result
Gastric infusion is after 4 weeks, compared with model group, and N1,N1-Dimethylbiguanide group, the fasting plasma glucose of the diabetic mice of hydroxytyrosol group and HT-NO group all has significance to reduce (P < 0.05), compared with hydroxytyrosol group, HT-NO group blood sugar decreasing effect is better, and result is as table 4.
Table 4HT-NO on the impact of blood glucose in diabetic mice (
n=8)
Note: compare with model control group: * P < 0.05
Embodiment 14
Hydroxytyrosol and the pharmacokinetic studies of HT-NO in rat body
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Medicine and reagent: HT (HPLC level purity >=98%), Ying Hua bio tech ltd, Xi'an; HT-NO (HPLC level purity >=95%), this Research team synthesizes; Quercetin (HPLC level purity >=99.3%), Shanghai is wildlife scientific & technical corporation forever; Methyl alcohol (chromatographic grade), Tianjin Ke Miou chemical reagents corporation; Ethyl acetate (AG), Tianjin Tian Li chemical reagents corporation; Formic acid, Guangdong brilliance chemical reagents corporation; Ultrapure water, TTL-30 ultrapure water system obtains.
Key instrument: Agilent1200 series of high efficiency liquid chromatograph, Shanghai WoWorkersScientificInstrumen; API3200 mass spectrograph, American AB SCIEX, FosterCity, CA company; N-EVAP112 type water bath with thermostatic control nitrogen dries up instrument, Organomation company of the U.S.; WH-2 micro-whirlpool vortex mixer, Shanghai Hu Xi analytical instrument company; TTL-30 ultrapure water system, Beijing is with safe tech equipment company limited; GENIUS16K-R low-temperature and high-speed whizzer, Changsha Xin Ao instrument company limited.
Laboratory animal: male SD rat, weight 250-300g, is provided by Xi'an Communications University's animal experimental center.
1.2 experimental techniques:
(1) set up the LC-MS/MS assay method of Hydroxytyrosol and HT-NO in rat plasma, and carry out Method validation;
(2) this method is applied to gavage Hydroxytyrosol and the pharmacokinetic studies of HT-NO in rat body.
1.3 experimental results:
(1) Method validation
1. linear and sensitivity: Hydroxytyrosol plasma concentration linearity range 0.00799-1.642 μ gmL
-1, linear relationship good (r=0.9984), lowest detectable limit (LLOQ) is 0.002 μ gmL
-1; HT-NO plasma concentration linearity range 0.00768-1.536 μ gmL
-1, linear relationship good (r=0.9990), lowest detectable limit (LLOQ) is 0.002 μ gmL
-1;
2. accuracy and precision: Hydroxytyrosol is in a few days respectively 2.30-3.88%, 2.34-5.32% with day to day precision RSD, in a few days and in the daytime accuracy is respectively 93.85-98.28%, 87.35-98.41%; Hydroxytyrosol NO derivative is in a few days respectively 2.84-4.69%, 3.14-6.15% with day to day precision RSD, and in a few days and in the daytime accuracy is respectively 90.12-98.44%, 85.77-98.86%;
3. extraction recovery: Hydroxytyrosol extraction recovery 87.17-97.94%, Hydroxytyrosol NO derivative extraction recovery is 86.40-101.14%.
4. stability: research shows, Hydroxytyrosol and HT-NO less stable, but the stability adding that 10 μ L1% xitix can significantly improve Hydroxytyrosol and NO derivative thereof.0.799 μ gmL
-1hydroxytyrosol plasma standard and 0.752 μ gmL
-1hT-NO plasma standard-20 DEG C of frozen stability of a week as shown in Figure 13-1,13-2.
(2) pharmacokinetic parameters
1. gastric infusion Hydroxytyrosol
Adopting 3P97 medicine to move software adopts one-compartment model to calculate pharmacokinetic parameters to Plasma Concentration result.The pharmacokinetic parameters of gavage Hydroxytyrosol the results are shown in Table 5.Hydroxytyrosol mean blood plasma concentration-time curve is shown in Figure 14.
Pharmacokinetic parameters (n=5) after table 5 gastric infusion HT in blood plasma
2. gastric infusion Hydroxytyrosol NO derivative
Adopting 3P97 medicine to move software adopts two-compartment model to calculate pharmacokinetic parameters to HT-NO Plasma Concentration result, adopts one-compartment model to calculate pharmacokinetic parameters to the Hydroxytyrosol Plasma Concentration result after administration HT-NO.After gavage HT-NO, its pharmacokinetic parameters the results are shown in Table 6, and mean blood plasma concentration-time curve is shown in Figure 15; After gavage HT-NO, Hydroxytyrosol pharmacokinetic parameters the results are shown in Table 7, and mean blood plasma concentration-time curve is shown in Figure 16;
The pharmacokinetic parameters of HT-NO in blood plasma (n=5) after table 6 gastric infusion HT-NO
The pharmacokinetic parameters of HT in blood plasma (n=5) after table 7 gastric infusion HT-NO
1.4 experiment conclusion
(1) this method is easy, sensitive, reproducible, can be applicable to the content of Hydroxytyrosol and HT-NO in mensuration blood plasma and is applied to gavage Hydroxytyrosol and the pharmacokinetic study of HT-NO in rat body.
(2) HT-NO can discharge Hydroxytyrosol and plays its pharmacology and physiological action by tachymetabolism in vivo.
Embodiment 15
Hydroxytyrosol and the HT-NO NO release in rat body is evaluated
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Reagent: NO test kit, Bioengineering Research Institute is built up in Nanjing, article No.: A012.
Key instrument: microplate reader; WH-2 micro-whirlpool vortex mixer, Shanghai Hu Xi analytical instrument company; GENIUS16K-R low-temperature and high-speed whizzer, Changsha Xin Ao instrument company limited; DK98-1 thermostat water bath, Tianjin Stettlen Instrument Ltd..
Laboratory animal: male SD rat, weight 250-300g, is provided by Xi'an Communications University's animal experimental center.
1.2 experimental technique
Rat is respectively after gavage Hydroxytyrosol and HT-NO, according to the time point of pharmacokinetic studies get rat artery blood centrifugal blood plasma, evaluate the dynamic level of active NO in 24h in rat plasma.Adopt the NO in Nitrate reductase mensuration rat plasma
2-, NO
3-, undertaken operating (test kit builds up Bioengineering Research Institute by Nanjing and provides) by test kit specification sheets, utilize microplate reader to measure, mensuration wavelength is 550nm.
1.3 experimental result
Gastric infusion 50mgkg respectively
-1hydroxytyrosol and 65mgkg
-1hT-NO after, in rat serum, in 24h, the dynamic level of NO is shown in Figure 17.
1.4 experiment conclusion
(1) HT-NO can discharge NO rapidly in rat body, makes it reach higher level.
(2) after administration Hydroxytyrosol in rat body NO level comparatively administration physiological saline is high, this is because Hydroxytyrosol can increase the synthesis of NO.
Claims (7)
1. a Hydroxytyrosol NO donor derivative, is characterized in that, its chemical structural formula is:
R is
wherein n=1 ~ 6.
2. the preparation method of a kind of Hydroxytyrosol NO donor derivative according to claim 1, is characterized in that:
As n=1, synthesis step is as follows:
1) 3,4-dihydroxybenzyl alcohols are substituted obtained 4-chloromethyl-1, the 2-dihydroxy-benzene of reaction;
2) 4-chloromethyl-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole;
3) 4-chloromethyl-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole;
4) 4-nitrooxymethyl-2,2-phenylbenzene benzo [1,3] dioxole obtains 4-nitrooxymethyl through acidic hydrolysis)-1,2-dihydroxy-benzene;
As n=2, synthesis step is as follows:
1) Hydroxytyrosol is substituted obtained 4-(2-chloroethyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(2-chloroethyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 5-(2-chloroethyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 5-(2-nitre trimethylammonium)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(2-nitre trimethylammonium)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=3, synthesis step is as follows:
1) 3,4-dihydroxy-benzene propyl alcohol are substituted obtained 4-(3-chloropropyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(3-chloropropyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 6-(3-chloropropyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 6-(3-nitre acryloxypropylethoxysilane)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(3-nitre acryloxypropylethoxysilane)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=4, synthesis step is as follows:
1) 3,4-dihydroxy-benzene butanols are substituted obtained 4-(4-chlorobutyl)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(4-chlorobutyl)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole;
3) 7-(4-chlorobutyl)-2,2-phenylbenzene benzo [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 7-(4-nitre acyloxy butyl)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(4-nitre acyloxy butyl)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=5, synthesis step is as follows:
1) 3,4-dihydroxy-benzene amylalcohols are substituted obtained 4-(5-chlorine amyl group)-1, the 2-dihydroxy-benzene of reaction;
2) 4-(5-chlorine amyl group)-1,2-dihydroxy-benzene and dichloro diphenyl methane react obtained 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
3) 8-(5-chlorine amyl group)-2,2-phenylbenzene benzos [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 8-(5-nitre acyloxy amyl group)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(5-nitre acyloxy amyl group)-1,2-dihydroxy-benzene through acidic hydrolysis;
As n=6, synthesis step is as follows:
1) obtained 4-(6-chlorine hexyl)-1, the 2-dihydroxy-benzene of reaction is substituted;
2) 4-(6-chlorine hexyl)-1,3,4-dihydroxy-benzene hexanol 2-dihydroxy-benzene and dichloro diphenyl methane react obtained 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
3) 9-(6-chlorine hexyl)-2,2-phenylbenzene benzos [1,3] dioxole and Silver Nitrate generation nucleophilic substitution reaction obtain 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole;
4) 9-(6-nitre acyloxy hexyl)-2,2-phenylbenzene benzos [1,3] dioxole obtains 4-(6-nitre acyloxy hexyl)-1,2-dihydroxy-benzene through acidic hydrolysis.
3. the application of a kind of Hydroxytyrosol NO donor derivative according to claim 1, is characterized in that: the application in the medicine preparing prevention and therapy diabetic vascular complications.
4. the application of a kind of Hydroxytyrosol NO donor derivative according to claim 3, is characterized in that: as the application of antioxidant.
5. the application of a kind of Hydroxytyrosol NO donor derivative according to claim 3, is characterized in that: the application in preparation vasodilation medicine.
6. the application of a kind of Hydroxytyrosol NO donor derivative according to claim 3, is characterized in that: protecting the application in medicine for the preparation of blood vessel endothelium.
7. the application of a kind of Hydroxytyrosol NO donor derivative according to claim 3, is characterized in that: preparing the application in hypoglycemic drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582108.0A CN105218377B (en) | 2015-09-14 | 2015-09-14 | A kind of hydroxytyrosol NO donor derivatives and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510582108.0A CN105218377B (en) | 2015-09-14 | 2015-09-14 | A kind of hydroxytyrosol NO donor derivatives and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105218377A true CN105218377A (en) | 2016-01-06 |
| CN105218377B CN105218377B (en) | 2017-10-27 |
Family
ID=54987722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510582108.0A Active CN105218377B (en) | 2015-09-14 | 2015-09-14 | A kind of hydroxytyrosol NO donor derivatives and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105218377B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1886416A (en) * | 2003-12-19 | 2006-12-27 | 弗·哈夫曼-拉罗切有限公司 | COMT inhibitors |
| CN103687843A (en) * | 2011-10-24 | 2014-03-26 | 尼科斯股份有限公司 | Quinone based nitric oxide donating compounds |
| WO2014063923A1 (en) * | 2012-10-23 | 2014-05-01 | Nicox S.A. | Quinone based nitric oxide donating compounds for ophthalmic use |
| WO2015044177A1 (en) * | 2013-09-24 | 2015-04-02 | Universität Zu Köln | Compounds useful in the treatment of neoplastic diseases |
-
2015
- 2015-09-14 CN CN201510582108.0A patent/CN105218377B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1886416A (en) * | 2003-12-19 | 2006-12-27 | 弗·哈夫曼-拉罗切有限公司 | COMT inhibitors |
| CN103687843A (en) * | 2011-10-24 | 2014-03-26 | 尼科斯股份有限公司 | Quinone based nitric oxide donating compounds |
| WO2014063923A1 (en) * | 2012-10-23 | 2014-05-01 | Nicox S.A. | Quinone based nitric oxide donating compounds for ophthalmic use |
| WO2015044177A1 (en) * | 2013-09-24 | 2015-04-02 | Universität Zu Köln | Compounds useful in the treatment of neoplastic diseases |
Non-Patent Citations (5)
| Title |
|---|
| DONATELLA BOSCHI等: "NO-Donor Phenols: A New Class of Products Endowed with Antioxidant and Vasodilator Properties", 《J.MED.CHEM.》 * |
| PAOLO TOSCO等: "Structure–Antioxidant Activity Relationships in a Series of NO-Donor Phenols", 《CHEMMEDCHEM》 * |
| SHIGEKI TAKAOKA等: "Isolation, synthesis, and neurite outgrowth-promoting activity of illicinin A from the flowers of Illicium anisatum", 《TETRAHEDRON》 * |
| WALFRED S.SAARI等: "Synthesis and Antihypertensive Activity of Some Ester Progenitors of Methyldopa", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
| 罗成，刘健康: "羟基酪醇作为线粒体营养素的调控机制", 《生命科学》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105218377B (en) | 2017-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100596299C (en) | Dehydrosilibinin diester derivatives, preparation method and use thereof | |
| CN101437529A (en) | Flavonoid compound having an antiviral activity | |
| Tang et al. | Design, synthesis and biological evaluation of novel xanthine oxidase inhibitors bearing a 2-arylbenzo [b] furan scaffold | |
| CN106632572B (en) | A kind of Astragaloside IV derivative and its preparation method and application | |
| Yang et al. | Antioxidant and α‐Glucosidase Inhibitory Activities Guided Isolation and Identification of Components from Mango Seed Kernel | |
| Meena et al. | α-Glucosidase inhibition activity and in silico study of 2-(benzo [d][1, 3] dioxol-5-yl)-4H-chromen-4-one, a synthetic derivative of flavone | |
| CN102285950B (en) | Application of amentoflavone | |
| CN106631804B (en) | A kind of compound and the preparation method and application thereof isolated from Labiatae Rabdosia plant | |
| CN105218377A (en) | A kind of Hydroxytyrosol NO donor derivative and its preparation method and application | |
| CN102786562B (en) | Pyrrolizidine alkaloids and purpose thereof | |
| CN102336790B (en) | Nitric oxide donator type giant knotweed glycoside derivates, preparation method and medical usage | |
| CN108752404B (en) | A kind of berberine salt derivative and its preparation method and application that triazole is sugar-modified | |
| CN107501367A (en) | The O β D glucuronic acids butyl esters of chrysoeriol 7 and its extracting method and purposes | |
| CN103044377B (en) | Compounds and composition with effects of inhibiting xanthine oxidase and reducing uric acid | |
| CN107446009A (en) | The O β D glucuronic acid methyl esters of chrysoeriol 7 and its extracting method and purposes | |
| CN101307090A (en) | Method for preparing timosaponin BIII and uses thereof | |
| CN106265681B (en) | A kind of triterpenoid is preparing the application in glycosidase inhibitor | |
| CN104987356A (en) | Ursolic acid-glycolysis inhibitor DCA conjugate and application thereof | |
| CN110452110A (en) | A kind of phloroglucinol derivatives natural drug and its preparation method and application | |
| CN103896957A (en) | Biphenyl furocoumarin compound and preparation method and application thereof | |
| CN103739653A (en) | Novel 23-oleanolic acid compound as well as preparation method and application of compound in preparation of glycosidase inhibitor medicine | |
| CN105012294A (en) | New uses of ellagic acid compound in preparation of hyperuricemia treating drug | |
| CN101829192B (en) | Artocarpus nitidus extract and use thereof | |
| CN106928292B (en) | A kind of nitrate NO donator type scutellarin derivative and its preparation method and application | |
| CN106565691B (en) | Benzo coumarin derivatives pharmaceutically acceptable salt and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |