CN105218377B - A kind of hydroxytyrosol NO donor derivatives and its preparation method and application - Google Patents
A kind of hydroxytyrosol NO donor derivatives and its preparation method and application Download PDFInfo
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- CN105218377B CN105218377B CN201510582108.0A CN201510582108A CN105218377B CN 105218377 B CN105218377 B CN 105218377B CN 201510582108 A CN201510582108 A CN 201510582108A CN 105218377 B CN105218377 B CN 105218377B
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Abstract
The invention discloses a kind of hydroxytyrosol NO donor derivatives and its preparation method and application, it is used to prepare the medicine in terms of hypoglycemic and blood vessel endothelium protection.Confirmed by inside and outside pharmacological evaluation, above-claimed cpd has following pharmacological activity：(1) scavenging capacity oxygen radical；(2) NO, vasodilator are discharged；(3) suppress glucosidase activity, there is certain hypoglycemic effect.Internal pharmacokinetics reach higher level test result indicates that it can discharge NO rapidly in vivo.The present invention provides the preparation method of hydroxytyrosol NO donor derivatives, is that can obtain changing derivative by 4 step organic syntheses, is easy to get with raw material, and reaction condition is gentle, and course of reaction is simple to operate, the cheap advantage of agents useful for same.The derivative has obvious hypoglycemic and vasorelaxation action, available for diabetes and its vascular complication is prevented and treated, with larger clinical value.
Description
Technical field：
The invention belongs to new drug development field, and in particular to a kind of hydroxytyrosol NO donor derivatives and preparation method thereof and
Application in terms of hypoglycemic and blood vessel endothelium protection.
Background technology：
Hydroxytyrosol (hydroxytyrosol, HT) chemical name Hydroxytyrosol, be have in olive oil it is strong anti-
In a kind of natural polyphenol class compound of oxidisability, the fruit and branches and leaves that are widely present in Oleaceae plants olive, to a variety of work
Property oxygen has elimination effect, with stronger antioxidation activity.In addition, numerous studies confirm hydroxytyrosol also in the prevention heart
Play the role of in terms of cranial vascular disease, anti-inflammatory anticancer good.Hydroxytyrosol is because of its special biology and pharmacological activity, extensively
Applied in food industry, nutrition and health care and cosmetics.The capsule and tablet that there have been hydroxytyrosol in foreign countries in succession at present come out.
Nitric oxide (NO) donator type medicine is to discharge NO through enzyme or non-enzyme effect in vivo.Classical medicine is that nitric acid is sweet
The single structures such as oil, sodium nitroprussiate directly discharge NO, pass through various linking groups and known drug or active pharmacophore and NO at present
Donor formation medicine attracts attention increasingly.It is proved：Angiosis can be obviously improved by giving NO substrates or exogenous NO donor medicines
Vasodilation and delay pathological process in change.What Merck companies of the U.S. were studied comes into II phase clinic NO- donator types COX-
2 inhibitor, exactly protect vascular endothelial cell, but also can reduce gastrointestinal tract mucous stimulation by discharging NO.Effect at present
It is to improve one of endothelial function effective ways in the NO donor medicines of NO links.
The content of the invention：
It is an object of the invention to provide a kind of hydroxytyrosol NO donor derivatives and preparation method thereof, and it is in reduction
Application in terms of blood glucose, anti-oxidation stress and reparation endothelial dysfunction.
To reach above-mentioned purpose, the present invention adopts the following technical scheme that to realize：
A kind of hydroxytyrosol NO donor derivatives, its chemical structural formula is：
R isWherein n=1~6.
A kind of preparation method of above-mentioned hydroxytyrosol NO donor derivatives, including：
As n=1, synthesis step is as follows：
1) 3,4- dihydroxybenzyl alcohols are substituted reaction and 4- chloromethyl -1,2- benzenediols are made；
2) reaction of 4- chloromethyls -1,2- benzenediols and dichloro diphenyl methane be made 5- chloromethyl -2,2- diphenyl benzo [1,
3] dioxole；
3) with silver nitrate nucleophilic substitution system occurs for 5- chloromethyls -2,2- diphenyl benzo [1,3] dioxole
Obtain 5- nitrooxymethyls -2,2- diphenyl benzo [1,3] dioxole；
4) 4- nitre is made by acidic hydrolysis in 5- nitrooxymethyls -2,2- diphenyl benzo [1,3] dioxole
Pivaloyloxymethyl) -1,2- benzenediols；
As n=2, synthesis step is as follows：
1) 3,4- dihydroxyphenyl ethanols are substituted reaction and 4- (2- chloroethyls) -1,2- benzenediols are made；
2) 5- (2- chloroethyls) -2,2- hexichol is made with dichloro diphenyl methane reaction in 4- (2- chloroethyls) -1,2- benzenediols
Base benzo [1,3] dioxole；
3) 5- (2- chloroethyls) -2,2- diphenyl benzo [1,3] dioxole and silver nitrate generation nucleophilic displacement of fluorine are anti-
5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole should be made；
4) 5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole is made by acidic hydrolysis
4- (2- nitre trimethylammonium) -1,2- benzenediols；
As n=3, synthesis step is as follows：
1) 3,4- dihydroxy benzenes propyl alcohol is substituted reaction and 4- (3- chloropropyls) -1,2- benzenediols is made；
2) 5- (3- chloropropyls) -2,2- hexichol is made with dichloro diphenyl methane reaction in 4- (3- chloropropyls) -1,2- benzenediols
Base benzo [1,3] dioxole；
3) 5- (3- chloropropyls) -2,2- diphenyl benzo [1,3] dioxole and silver nitrate generation nucleophilic displacement of fluorine are anti-
5- (3- nitre acryloxypropylethoxysilane) -2,2- diphenyl benzo [1,3] dioxole should be made；
4) 5- (3- nitre acryloxypropylethoxysilane) -2,2- diphenyl benzo [1,3] dioxole is made by acidic hydrolysis
4- (3- nitre acryloxypropylethoxysilane) -1,2- benzenediols；
As n=4, synthesis step is as follows：
1) 3,4- dihydroxy benzenes butanol is substituted reaction and 4- (4- chlorobutyls) -1,2- benzenediols is made；
2) 5- (4- chlorobutyls) -2,2- hexichol is made with dichloro diphenyl methane reaction in 4- (4- chlorobutyls) -1,2- benzenediols
Base benzo [1,3] dioxole；
3) 5- (4- chlorobutyls) -2,2- diphenyl benzo [1,3] dioxole and silver nitrate generation nucleophilic displacement of fluorine are anti-
5- (4- nitryl epoxides butyl) -2,2- diphenyl benzo [1,3] dioxole should be made；
4) 5- (4- nitryl epoxides butyl) -2,2- diphenyl benzo [1,3] dioxole is made by acidic hydrolysis
4- (4- nitryl epoxides butyl) -1,2- benzenediols；
As n=5, synthesis step is as follows：
1) 3,4- dihydroxy benzenes amylalcohol is substituted reaction and 4- (5- chlorine amyl group) -1,2- benzenediols is made；
2) 5- (5- chlorine amyl group) -2,2- hexichol is made with dichloro diphenyl methane reaction in 4- (5- chlorine amyl group) -1,2- benzenediols
Base benzo [1,3] dioxole；
3) 5- (5- chlorine amyl group) -2,2- diphenyl benzo [1,3] dioxole and silver nitrate generation nucleophilic displacement of fluorine are anti-
5- (5- nitryl epoxides amyl group) -2,2- diphenyl benzo [1,3] dioxole should be made；
4) 5- (5- nitryl epoxides amyl group) -2,2- diphenyl benzo [1,3] dioxole is made by acidic hydrolysis
4- (5- nitryl epoxides amyl group) -1,2- benzenediols；
As n=6, synthesis step is as follows：
1) it is substituted reaction and 4- (6- chlorine hexyl) -1,2- benzenediols is made；
2) 5- (6- chlorine hexyl) -2,2- hexichol is made with dichloro diphenyl methane reaction in 4- (6- chlorine hexyl) -1,2- benzenediols
Base benzo [1,3] dioxole；
3) 5- (6- chlorine hexyl) -2,2- diphenyl benzo [1,3] dioxole and silver nitrate generation nucleophilic displacement of fluorine are anti-
5- (6- nitryl epoxides hexyl) -2,2- diphenyl benzo [1,3] dioxole should be made；
4) 5- (6- nitryl epoxides hexyl) -2,2- diphenyl benzo [1,3] dioxole is made by acidic hydrolysis
4- (6- nitryl epoxides hexyl) -1,2- benzenediols.
A kind of application of above-mentioned hydroxytyrosol NO donor derivatives, is preparing prevention and treatment diabetic vascular complications
Application in medicine.
Further improve of the invention is：It is used as the application of antioxidant.
Further improve of the invention is：Application in blood vessel dilatation medicine is prepared.
Further improve of the invention is：Preparing the application in protecting medicine for blood vessel endothelium.
Further improve of the invention is：Application in hypoglycemic drug is prepared.
Relative to prior art, the present invention has the advantage that：
The present invention is, using natural polyphenol class compound hydroxytyrosol as parent nucleus, different length to be introduced on its 4 side chains
Carbochain, synthesizes a series of hydroxytyrosol NO donor derivatives.It can be obtained, be easy to get with raw material by 4 step organic syntheses, react bar
Part is gentle, and course of reaction is simple to operate, the cheap advantage of agents useful for same.The present invention is confirmed by inside and outside pharmacological evaluation, above-mentioned
Compound has following pharmacological activity：(1) scavenging capacity oxygen radical；(2) NO, vasodilator are discharged；(3) glucoside is suppressed
Enzymatic activity, there is certain hypoglycemic effect.Internal pharmacokinetics are reached test result indicates that it can discharge NO rapidly in vivo
Higher level.
The hydroxytyrosol NO donor derivatives that the present invention is provided have good hypoglycemic and vasorelaxation action, available for preventing
Diabetes and its vascular complication are controlled, with larger clinical value.
Brief description of the drawings：
Fig. 1 is the synthetic route chart of 4- (2- nitrooxymethyls) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 2 is the synthetic route chart of 4- (2- nitre trimethylammonium) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 3 is the synthetic route chart of 4- (2- nitre acryloxypropylethoxysilane) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 4 is the synthetic route chart of 4- (2- nitryl epoxides butyl) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 5 is the synthetic route chart of 4- (2- nitryl epoxides amyl group) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 6 is the synthetic route chart of 4- (2- nitryl epoxides hexyl) -1,2- benzenediols；
Marked in figure be specially：
(a)CCl4, PPh3, MeCN；(b)Ph2CCl2, PhMe；(c)AgNO3, MeCN；(d)AcOH：H2O(4:1)。
Fig. 7 is the external NO release profiles of HT-NO.
Fig. 8 is influences of the HT-NO to rat chest aorta ring diastole activity.
Fig. 9 is influences of the HT-NO to Human umbilical vein endothelial cells activity.
Figure 10 is influences of the HT-NO to the Human umbilical vein endothelial cells activity of high sugar induced.
Figure 11 is influences of the HT-NO to Human umbilical vein endothelial cells NO levels.
Figure 12 is influences of the HT-NO to the Human umbilical vein endothelial cells NO levels of high sugar induced.
Figure 13-1 is that HT adds the concentration curve that 1% ascorbic acid freezes one week under the conditions of-20 DEG C.
Figure 13-2 is that HT-NO adds the concentration curve that 1% ascorbic acid freezes one week under the conditions of-20 DEG C.
Figure 14 is rat oral gavage HT blood concentration-time curve.
Figure 15 is rat oral gavage HT-NO blood concentration-time curve.
Figure 16 is the blood concentration-time curve of HT after rat oral gavage HT-NO.
Figure 17 is the horizontal change curves of NO in blood plasma in rat oral gavage HT-NO, HT and physiological saline 24h.
Embodiment：
Below used embodiment come the invention will be further described, it should be pointed out that be these embodiments be only used for this
Invention is illustrated, and should not be construed as limiting the invention.
HT-NO synthesis step：
Embodiment 1
The synthesis of 4- chloromethyl -1,2- benzenediols
3,4- dihydroxybenzyl alcohols are dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- chloromethanes
Base -1,2- benzenediols；
The synthesis of 5- chloromethyls -2,2- diphenyl benzo [1,3] dioxole
4- chloromethyl -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, return stirring is added
24h, after question response is complete, is cooled to room temperature, reaction system H2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3),
Merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- chloromethyls -2,2-
Diphenyl benzo [1,3] dioxole；
The synthesis of 5- nitrooxymethyls -2,2- diphenyl benzo [1,3] dioxole
5- chloromethyl -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, nitre is added
Acid is silver-colored (2.50g), under nitrogen protection, lucifuge, and stirring extremely reaction completely, adds NaCl solution and 1h is stirred at room temperature, filter at 57 DEG C
Go out solid, acetonitrile solvent is removed under reduced pressure, add H2O (20mL), is extracted with ethyl acetate (20mL × 3), merges organic phase, nothing
Aqueous sodium persulfate is dried, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- nitrooxymethyl -2,2- diphenyl
Benzo [1,3] dioxole；
The synthesis of 4- nitrooxymethyl -1,2- benzenediols
5- nitrooxymethyls -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O(20mL/
In mixed solution 5mL), under nitrogen protection, return stirring 4-5h after question response is complete, is cooled to room temperature, adds ethyl acetate
(50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying, filtering,
Be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- nitrooxymethyl -1,2- benzenediols.
Embodiment 2
The synthesis of 4- (2- chloroethyls) -1,2- benzenediols
Hydroxytyrosol is dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (2- chlorine
Ethyl) -1,2- benzenediols；
The synthesis of 5- (2- chloroethyls) -2,2- diphenyl benzo [1,3] dioxole
4- (2- chloroethyls) -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, backflow is added
24h is stirred, after question response is complete, room temperature, reaction system H is cooled to2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3)
Take, merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (2- chloroethenes
Base) -2,2- diphenyl benzo [1,3] dioxole；
The synthesis of 5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole
5- (2- chloroethyls) -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, plus
Enter silver nitrate (2.50g), under nitrogen protection, lucifuge, stirring adds NaCl solution and 1h is stirred at room temperature to reacting complete at 57 DEG C,
Solid is filtered away, acetonitrile solvent is removed under reduced pressure, H is added2O (20mL), is extracted with ethyl acetate (20mL × 3), is merged organic
Phase, anhydrous sodium sulfate drying, filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (2- nitre trimethylammonium) -2,
2- diphenyl benzo [1,3] dioxole；
The synthesis of 4- (2- nitre trimethylammonium) -1,2- benzenediols：
5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O
In the mixed solution of (20mL/5mL), under nitrogen protection, after return stirring 4-5h, but reaction completely, room temperature is cooled to, second is added
Acetoacetic ester (50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying,
Filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (2- nitre trimethylammonium) -1,2- benzenediols.
Embodiment 3
The synthesis of 4- (3- chloropropyls) -1,2- benzenediols
3,4- dihydroxy benzenes propyl alcohol is dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (3- chlorine
Propyl group) -1,2- benzenediols；
The synthesis of 5- (3- chloropropyls) -2,2- diphenyl benzo [1,3] dioxole
4- (3- chloropropyls) -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, backflow is added
24h is stirred, after question response is complete, room temperature, reaction system H is cooled to2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3)
Take, merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (3- chlorine third
Base) -2,2- diphenyl benzo [1,3] dioxole；
The synthesis of 5- (3- nitre acryloxypropylethoxysilane) -2,2- diphenyl benzo [1,3] dioxole
5- (3- chloropropyls) -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, plus
Enter silver nitrate (2.50g), under nitrogen protection, lucifuge, stirring adds NaCl solution and 1h is stirred at room temperature to reacting complete at 57 DEG C,
Solid is filtered away, acetonitrile solvent is removed under reduced pressure, H is added2O (20mL), is extracted with ethyl acetate (20mL × 3), is merged organic
Phase, anhydrous sodium sulfate drying, filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (3- nitre acryloxypropylethoxysilane) -2,
2- diphenyl benzo [1,3] dioxole；
The synthesis of 4- (3- nitre acryloxypropylethoxysilane) -1,2- benzenediols：
5- (3- nitre acryloxypropylethoxysilane) -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O
In the mixed solution of (20mL/5mL), under nitrogen protection, after return stirring 4-5h, but reaction completely, room temperature is cooled to, second is added
Acetoacetic ester (50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying,
Filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (3- nitre acryloxypropylethoxysilane) -1,2- benzenediols.
Embodiment 4
The synthesis of 4- (4- chlorobutyls) -1,2- benzenediols
3,4- dihydroxy benzenes butanol is dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (4- chlorine
Butyl) -1,2- benzenediols；
The synthesis of 5- (4- chlorobutyls) -2,2- diphenyl benzo [1,3] dioxole
4- (4- chlorobutyls) -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, backflow is added
24h is stirred, after question response is complete, room temperature, reaction system H is cooled to2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3)
Take, merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (4- neoprenes
Base) -2,2- diphenyl benzo [1,3] dioxole；
The synthesis of 5- (4- nitryl epoxides butyl) -2,2- diphenyl benzo [1,3] dioxole
5- (4- chlorobutyls) -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, plus
Enter silver nitrate (2.50g), under nitrogen protection, lucifuge, stirring adds NaCl solution and 1h is stirred at room temperature to reacting complete at 57 DEG C,
Solid is filtered away, acetonitrile solvent is removed under reduced pressure, H is added2O (20mL), is extracted with ethyl acetate (20mL × 3), is merged organic
Phase, anhydrous sodium sulfate drying, filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (4- nitryl epoxides butyl) -2,
2- diphenyl benzo [1,3] dioxole；
The synthesis of 4- (4- nitryl epoxides butyl) -1,2- benzenediols：
5- (4- nitryl epoxides butyl) -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O
In the mixed solution of (20mL/5mL), under nitrogen protection, after return stirring 4-5h, but reaction completely, room temperature is cooled to, second is added
Acetoacetic ester (50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying,
Filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (4- nitryl epoxides butyl) -1,2- benzenediols.
Embodiment 5
The synthesis of 4- (5- chlorine amyl group) -1,2- benzenediols
3,4- dihydroxy benzenes amylalcohols are dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (5- chlorine
Amyl group) -1,2- benzenediols；
The synthesis of 5- (5- chlorine amyl group) -2,2- diphenyl benzo [1,3] dioxole
4- (5- chlorine amyl group) -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, backflow is added
24h is stirred, after question response is complete, room temperature, reaction system H is cooled to2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3)
Take, merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (5- chlorine penta
Base) -2,2- diphenyl benzo [1,3] dioxole；
The synthesis of 5- (5- nitryl epoxides amyl group) -2,2- diphenyl benzo [1,3] dioxole
5- (5- chlorine amyl group) -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, plus
Enter silver nitrate (2.50g), under nitrogen protection, lucifuge, stirring adds NaCl solution and 1h is stirred at room temperature to reacting complete at 57 DEG C,
Solid is filtered away, acetonitrile solvent is removed under reduced pressure, H is added2O (20mL), is extracted with ethyl acetate (20mL × 3), is merged organic
Phase, anhydrous sodium sulfate drying, filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (5- nitryl epoxides amyl group) -2,
2- diphenyl benzo [1,3] dioxole；
The synthesis of 4- (5- nitryl epoxides amyl group) -1,2- benzenediols：
5- (5- nitryl epoxides amyl group) -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O
In the mixed solution of (20mL/5mL), under nitrogen protection, after return stirring 4-5h, but reaction completely, room temperature is cooled to, second is added
Acetoacetic ester (50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying,
Filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (5- nitryl epoxides amyl group) -1,2- benzenediols.
Embodiment 6
The synthesis of 4- (6- chlorine hexyl) -1,2- benzenediols
3,4- dihydroxy benzenes hexanols are dissolved in 25mL anhydrous acetonitriles, triphenyl phosphorus, carbon tetrachloride, nitrogen is sequentially added
Protection, is stirred at room temperature 10h, after question response is complete, and acetonitrile solvent is removed under reduced pressure, and adds H2O (20mL), is extracted with dichloromethane,
Saturation NaCl solution is washed, anhydrous sodium sulfate drying, filtering, and be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (6- chlorine
Hexyl) -1,2- benzenediols；
The synthesis of 5- (6- chlorine hexyl) -2,2- diphenyl benzo [1,3] dioxole
4- (6- chlorine hexyl) -1,2- benzenediols are dissolved in toluene, dichloro diphenyl methane, nitrogen protection, backflow is added
24h is stirred, after question response is complete, room temperature, reaction system H is cooled to2O (50mL) is washed, and aqueous phase is extracted with toluene (20mL × 3)
Take, merge organic phase, anhydrous sodium sulfate drying is filtered, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtain 5- (6- chlorine oneself
Base) -2,2- diphenyl benzo [1,3] dioxole；
The synthesis of 5- (6- nitryl epoxides hexyl) -2,2- diphenyl benzo [1,3] dioxole
5- (6- chlorine hexyl) -2,2- diphenyl benzo [1,3] dioxole is dissolved in 25mL anhydrous acetonitriles, plus
Enter silver nitrate (2.50g), under nitrogen protection, lucifuge, stirring adds NaCl solution and 1h is stirred at room temperature to reacting complete at 57 DEG C,
Solid is filtered away, acetonitrile solvent is removed under reduced pressure, H is added2O (20mL), is extracted with ethyl acetate (20mL × 3), is merged organic
Phase, anhydrous sodium sulfate drying, filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 5- (6- nitryl epoxides hexyl) -2,
2- diphenyl benzo [1,3] dioxole；
The synthesis of 4- (6- nitryl epoxides hexyl) -1,2- benzenediols：
5- (6- nitryl epoxides hexyl) -2,2- diphenyl benzo [1,3] dioxole is added to AcOH/H2O
In the mixed solution of (20mL/5mL), under nitrogen protection, after return stirring 4-5h, but reaction completely, room temperature is cooled to, second is added
Acetoacetic ester (50mL) and H2O (50mL), organic phase is washed with saturated solution of sodium bicarbonate (20mL × 3), anhydrous sodium sulfate drying,
Filtering, be concentrated under reduced pressure to obtain yellow oil.Column chromatography for separation obtains 4- (6- nitryl epoxides hexyl) -1,2- benzenediols.
Embodiment 7
HT-NO antioxidation activities are tested
Using free radical method ([DPPH] method) is removed, to 4- (2- nitre trimethylammonium) -1,2- benzenediol (following shorthands
For HT-NO) antioxidation activity through row test.
1 materials and methods
1.1 Experimental agents
Embodiment 2 prepares gained compound
Experiment material：
Reagent：
1,1- diphenyl -2- trinitrophenyl-hydrazines (1,1-Diphenyl-2-picrylhydrazyl [DPPH] (upper Hypons
Graceful bio tech ltd)；Absolute ethyl alcohol (Tianjin Tian Li chemical reagent Co., Ltd)；
Key instrument：752 type ultraviolet-uisible spectrophotometers (Shanghai Spectrum Apparatus Co., Ltd.)；WH-2 micro-whirlpools are mixed
Close instrument (Shanghai Hu Xi analytical instrument Co., Ltd., Factory)；
1.2 experimental method
The preparation of detected sample：
1) [DPPH] and sample sets (Ai)：It is 0.08mg/mL that 2mL concentration is added in 10mL tool edge test tube
[DPPH] ethanol solution, being separately added into the testing sample (HT-NO) and positive control of 2mL various concentrations, (0.1mg/mL is anti-bad
Hematic acid)；
2) [DPPH] and solvent group (Ao)：It is 0.08mg/mL that 2mL concentration is added in 10mL tool edge test tube
[DPPH] ethanol solution and 2mL absolute ethyl alcohols；
3) sample and solvent group (Aj)：2mL absolute ethyl alcohols are added in 10mL tool edge test tube, 2mL are separately added into different
The testing sample (HT-NO) and positive control (0.1mg/mL ascorbic acid) of concentration；
After detected sample sample-adding is finished, shake up, be stored at room temperature 30min, light absorption value is detected in 517nm wavelength.According to
Following formula calculates free radical scavenging activity.
[DPPH] free radical scavenging activity S%=[1- (Ai-Aj)/Ao] × 100%.
1.3 experimental result
Using SPSS matched curves and calculate IC50, HT-NO is obtained to [DPPH] half elimination ratio IC50For 3.28 × 10- 5mol·L-1, ascorbic acid is to [DPPH] half elimination ratio IC50For 6.95 × 10-5mol·L-1.As a result show, HT-NO tools
There is good radical scavenging activity.As a result such as table 1, table 2
Table 1 [DPPH] method determines HT-NO antioxidation activity result (%)
Table 2 [DPPH] method determines the antioxidation activity result (%) of ascorbic acid
Embodiment 8
The external NO releases activity experiments of HT-NO
Activity is discharged to HT-NO external NO through row test using Griess methods.
1 materials and methods
1.1 Experimental agents
Embodiment 2 prepares gained compound
Experiment material：
Reagent：Anhydrous p-aminobenzene sulfonic acid is received (Tianjin good fortune morning chemical reagent factory)；N-1- naphthodiamide hydrochlorides (Tianjin
Loftily fine chemistry industry research institute)；Natrium nitrosum (Tianjin good fortune morning chemical reagent factory)；Cys (Tianjin good fortune morning chemistry
Chemical reagent work)；Potassium dihydrogen phosphate (Guangdong Guanghua Science and Technology Co., Ltd.)；Sodium hydroxide (Tianjin Hedong District red rock reagent
Factory)；Phosphoric acid (Xi'an chemical reagent factory).
Key instrument：752 type ultraviolet-uisible spectrophotometers (Shanghai Spectrum Apparatus Co., Ltd.'s manufacture)；The miniature whirlpools of WH-2
Revolve mixed instrument (manufacture of Shanghai Hu Xi analytical instrument Co., Ltd., Factory)；
1.2 experimental method
(1) drafting of NO release standards curve：Phosphate buffer solution 2.5mL, methanol 2.5mL, isometric mixing, then add
Enter the NaNO of various concentrations2 -With 50 μ L 0.1mol/L Cys solution, Griess reagents are added after mixing, are mixed quiet
10min is put, in determining its absorbance under 540nm.Using SPSS fit standard curves, and standard curve is obtained by curve matching
Equation.
(2) HT-NO external NO releases are determined：Phosphate buffer solution 2.5mL, methanol 2.5mL, isometric mixing,
The HT-NO and 50 μ L of various concentrations 0.1mol/L Cys solution are added, Griess reagents are added after mixing, is mixed
10min is stood, in determining its absorbance under 540nm.Obtained accordingly using SPSS matched curves, and by calibration curve equation
NO2 -Concentration, sodium nitroprussiate and hydroxytyrosol derivatives release NO efficiency=NO release concentrations derive with 0.1mM in initial reaction liquid
Thing contains the ratio of NO molal quantity.
1.3 experimental result
As a result show that HT-NO can discharge certain density NO in vitro.As a result such as Fig. 7
Embodiment 9
Influence experiments of the HT-NO to Endothelial Function
1 materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Sodium nitroprussiate, SD rats, computer bio signal record analysis system, tonotransducer, in vitro tissue organ constant temperature
Perfusion system.
1.2 experimental method
Cervical dislocation puts to death rat, and sustainer is taken out rapidly, is placed in 4 DEG C of tyrode's solutions for being connected with mixed gas, rejects blood
The fat and connective tissue of pipe outer wall.Vascular strip is cut into segment, every section of 2~3mm or so；One section of vascular strip is taken, through fixation
Spillikin, then blood vessel is hung up with small hook, small hook is connected to sensor with thin cotton thread.Finally by blood vessel be soaked in equipped with tyrode's solution,
It is connected with mixed gas and keeps in the reaction tank of 37 DEG C of constant temperature.By software, the state of tension of vascular strip is monitored, by poised state
Pulling force adjust to 0.8g or so；1h is balanced, tyrode's solution is changed 1 time per 15min.Stimulated 3 times with 60mmol KCl, treat that blood vessel is received
When being reduced to most strong and stable, the HT-NO of various concentrations is added, using physiological saline as control, to add the blood after medicine 15min
Ratio between the maximum shrinkage amplitude for the vascular circle that pipe tension force amplitude and KCl induce reflects the change of antiotasis.
1.3 experimental result
As a result show that HT-NO can suppress KCl to contraction caused by vascular circle.As a result such as Fig. 8
Embodiment 10
Influences of the HT-NO to the Human umbilical vein endothelial cells activity of high sugar induced is tested
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
DMEM culture medium dry powders (Gibco, USA), hyclone (FBS) (Gibco, USA), pancreatin (Trypsin)
(Beyotime), MTT (Sigma), glucose (Sigma), human umbilical vein endothelial (HUVECs), ELIASA (Thermo
Scientific)
1.2 experimental method
1.2.1 cell culture：With DMEM (Dulbecco ' the s Modified Eagle ' s Medium) trainings containing 10%FBS
Nutrient solution culture HUVECs cells, were passed on 1 time per 1-2 days.
1.2.2 HUVECs cells are inoculated in 96 orifice plates in units of 5000-6000/hole, are divided into blank control group, it is high
Sugared group and administration group.Model group gives 55mM glucose solution, administration group give various concentrations HT-NO (12.5,25,50,
100th, 200 μM) and 55mM height sugar common be incubated 48h.After incubation terminates, 20 μ L MTT are added per hole, 4h is incubated, it is molten with DMSO
Absorbance (A) is determined after solution under ELIASA 492nm wavelength.Cytoactive (%) is calculated as follows：Cytoactive (%)
=(A experimental group-A blank groups)/(A control group-A blank groups) × 100%, calculates corresponding cytoactive.
1.3 experimental result
As a result show and individually give HT-NO effect endothelial cells, cell can be promoted to breed in the range of finite concentration, such as
Fig. 9；And give after the high sugar stimulation of endothelial cell, cytoactive is substantially reduced, and giving HT-NO can damage to the cell of high sugar induced
Wound plays certain protective effect, such as Figure 10.
Embodiment 11
Influence experiments of the HT-NO to the Human umbilical vein endothelial cells NO levels of high sugar induced
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
DMEM culture medium dry powders (Gibco, USA), hyclone (FBS) (Gibco, USA), pancreatin (Trypsin)
(Beyotime), MTT (Sigma), glucose (Sigma), human umbilical vein endothelial (HUVECs), ELIASA (Thermo
Scientific)
1.2 experimental method
1.2.1 cell culture：With DMEM (Dulbecco ' the s Modified Eagle ' s Medium) trainings containing 10%FBS
Nutrient solution culture HUVECs cells, were passed on 1 time per 1-2 days.
1.2.2 HUVECs cells are inoculated in 96 orifice plates in units of 5000-6000/hole, are divided into blank control group, it is high
Sugared group and administration group.Model group gives 55mM glucose solution, administration group give various concentrations HT-NO (12.5,25,50,
100th, 200 μM) and 55mM height sugar common be incubated 48h.After incubation terminates, cell supernatant is collected, is existed with NO detection kits
Absorbance (A) is determined under ELIASA 550nm wavelength.Corresponding NO concentration is calculated by detection kit specification.
1.3 experimental result
As a result show and individually give HT-NO effect endothelial cells, cell supernatant can be promoted in the range of finite concentration
Middle NO release, such as Figure 11；And give after the high sugar stimulation of endothelial cell, NO contents are reduced in cell supernatant, and giving HT-NO can
NO level, such as Figure 12 in cell supernatant to raise high sugar induced.
Embodiment 12
HT-NO external glucoside inhibiting activity experiment
1 materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Acarbose (Bayer A.G, 50mg/ pieces are commercially available), (bioengineering is built up in Nanjing to glucose determination reagent box
Research institute), alpha-glucosidase (Sigma), 4- nitrobenzene-α-D- glucopyranosides (α-PNPG) (Sigma), 752 types are purple
Outer visible spectrophotometer (Shanghai Spectrum Apparatus Co., Ltd.'s manufacture)
1.2 experimental method
1.2.1 yeast α-glucosidase is suppressed
(1) α-PNPG solution is prepared to prepare；Alpha-glucosaccharase enzyme solutions 10U/mL；NaCO3 solution 1mol/L；Various concentrations
Target compound prepare liquid (with volume ratio be 1:9 DMSO- phosphate buffers dissolving).
(2) the μ L of alpha-glucosaccharase enzyme solutions 10 are taken, 10min is incubated in 37 DEG C of water-baths, is separately added into treating for various concentrations
The μ L of solution 50 are surveyed, follow-up continuation of insurance temperature 10min is mixed, adds the μ L of α-PNPG solution 20,15min is incubated after mixing, NaCO3 is added
The μ L terminating reactions of solution 50.Blank control (without enzyme and sample) and negative control group (being free of sample) are set up, acarbose is
Positive control.Solution absorbance is detected at 405nm, inhibiting rate (%) is calculated as follows：Inhibiting rate (%)=(A is negative
Control-A samples)/(A negative control-A blank controls) × 100%, calculate corresponding half-inhibition concentration (IC50)。
1.2.2 small intestine in rats alpha-glucosidase is suppressed
(1) preparation of small intestine in rats enzyme liquid：Rat Fast, which can't help taking off cervical vertebra after water 12h, puts to death, and small intestine is taken out, with cold
Normal saline flushing enteric cavity, clean small intestine is placed on ice platform, is scraped mucous membrane of small intestine, is weighed.Add 4 DEG C of phosphorus of 5 times of amounts
Phthalate buffer, is homogenized in ice bath, and then 5000r/min centrifuges 20min, takes supernatant.
(2) the μ L of enzyme liquid 10 are taken, are incubated in 37 DEG C of water-baths after 15min, the μ L of inhibitor 50 of various concentrations are added, continue to protect
Warm 30min, adds the 50mmol/L μ L of maltose 20, is reacted in 37 DEG C of water-baths after 60min, and taking-up is placed in boiling water bath eventually
Only react；Blank control (without inhibitor) is done simultaneously, acarbose is positive control.10 μ are extracted reaction solution in EP pipes, then are added
Enter in glucose kit the μ L of 500 μ L, B test solution of A test solutions 500 to mix, after 37 DEG C of insulation 15min, in determining its extinction under 505nm
Degree.Inhibiting rate (%) is calculated as follows：Inhibiting rate (%)=(A blank control-A samples)/A blank control × 100%, meter
Calculate corresponding half-inhibition concentration (IC50)。
1.3 experimental result
As a result yeast α-glucosidase and small intestine in rats alpha-glucosidase can be suppressed by showing the compound of preparation.Knot
Fruit such as table 3
Inhibitory action ICs of the HT-NO of table 3 to alpha-glucosidase50(μM)
Embodiment 13
Influence experiments of the HT-NO to the STZ blood glucose in diabetic mice induced
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Reagent：STZ (Sigma), melbine bulk drug (Tokyo HuaCheng Industry Co., Ltd), hydroxytyrosol (Shaanxi should
Change Bioisystech Co., Ltd), the stable blood glucose meter of three promises and blood sugar test paper bar (Sinocare Biosensing Co., Ltd)
Experimental animal：Kunming mice, weight 25-30g, by Xi'an Communications University, animal experimental center is provided.
1.2 experimental method
1.2.1 diabetic mouse model is set up
After 50 mouse adaptability of purchase are fed 3 days, 8 are therefrom randomly selected as Normal group, is given
Chow diet is fed, and then remaining 42 overnight fasting gives experiment mice abdominal cavity injection location a certain amount of STZ citric acids-lemon
Sour sodium (160mgkg-1).The citric acid-sodium citrate buffer of same dose volume also gives the mouse of Normal group
Injection.All equal fasting of mouse in the 5th day after injection, the mouse tail vein of modeling takes blood to measure its fasting blood-glucose, empty with mouse
Abdomen blood glucose value >=16.7mmolL-1For the successful standard of diabetic experimental mouse model.
1.2.2 experimental animal packet and administration
The successful mouse of modeling is randomly divided into：Model group, melbine group (250mgkg-1), hydroxytyrosol group
(77mg·kg-1) and HT-NO groups (100mgkg-1), every group of mouse 8.Continuous gavage 4 weeks, normal group and model group are given
The physiological saline gavage of equal volume.The visual condition of daily observation mouse, while recording each group amount of drinking water and dietary amount.
1.3 experimental result
After gastric infusion 4 weeks, compared with model group, melbine group, the diabetes of hydroxytyrosol group and HT-NO groups are small
The fasting blood-glucose of mouse has conspicuousness to reduce (P ＜ 0.05), compared with hydroxytyrosol group, and HT-NO groups blood sugar decreasing effect more preferably, is tied
Fruit such as table 4.
Influences of the HT-NO of table 4 to blood glucose in diabetic mice
Note：Compared with model control group：* P ＜ 0.05
Embodiment 14
The pharmacokinetic studies of hydroxytyrosol and HT-NO in rat body
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Medicine and reagent：HT (HPLC grades of purity >=98%), Xi'an Ying Hua bio tech ltd；(HPLC grades of HT-NO
Purity >=95%), the synthesis of this Research team；Quercetin (HPLC grades of purity >=99.3%), Shanghai wildlife scientific ＆ technical corporation forever；First
Alcohol (chromatographic grade), Tianjin Ke Miou chemical reagents corporations；Ethyl acetate (analysis level), Tianjin Tian Li chemical reagents corporations；Formic acid,
Guangdong brilliance chemical reagents corporation；Ultra-pure water, TTL-30 ultrapure water systems are obtained.
Key instrument：The series of high efficiency liquid chromatographs of Agilent 1200, Shanghai WoWorkers Scientific
Instrumen；API3200 mass spectrographs, American AB SCIEX, Foster City, CA companies；N-EVAP112 type water bath with thermostatic control nitrogen
The dry instrument of air-blowing, Organomation companies of the U.S.；WH-2 micro-whirlpool vortex mixers, Shanghai Hu Xi analytical instrument company；TTL-30
Ultrapure water system, Beijing is with safe tech equipment Co., Ltd；GENIUS 16K-R low-temperature and high-speed centrifuges, the prosperous instrument instrument difficult to understand in Changsha
Table Co., Ltd.
Experimental animal：Male SD rat, weight 250-300g, by Xi'an Communications University, animal experimental center is provided.
1.2 experimental method：
(1) the LC-MS/MS determination methods of hydroxytyrosol and HT-NO in rat plasma are set up, and carry out Method validation；
(2) the method is applied to the pharmacokinetic studies of gavage hydroxytyrosol and HT-NO in rat body.
1.3 experimental result：
(1) Method validation
1. linear and sensitivity：Hydroxytyrosol plasma concentration range of linearity 0.00799-1.642 μ gmL-1, linear relationship
Well (r=0.9984), minimum detection limit (LLOQ) is 0.002 μ gmL-1；HT-NO plasma concentration ranges of linearity 0.00768-
1.536μg·mL-1, well (r=0.9990), minimum detection limit (LLOQ) is 0.002 μ gmL to linear relationship-1；
2. accuracy and precision：Hydroxytyrosol is respectively in a few days 2.30-3.88%, 2.34- with day to day precision RSD
5.32%, in a few days and in the daytime accuracy is respectively 93.85-98.28%, 87.35-98.41%；Hydroxytyrosol NO derivatives are in a few days
It is respectively 2.84-4.69%, 3.14-6.15% with day to day precision RSD, in a few days and in the daytime accuracy is respectively 90.12-
98.44%th, 85.77-98.86%；
3. extraction recovery：Hydroxytyrosol extraction recovery 87.17-97.94%, hydroxytyrosol NO derivative, which are extracted, to be reclaimed
Rate is 86.40-101.14%.
4. stability：Research shows, hydroxytyrosol and HT-NO less stables, but adds 10 μ L1% ascorbic acid energy
Enough significantly improve the stability of hydroxytyrosol and its NO derivatives.0.799μg·mL-1Hydroxytyrosol plasma standard and 0.752 μ
g·mL-1HT-NO plasma standards freeze the stability of one week at-20 DEG C as shown in Figure 13-1,13-2.
(2) pharmacokinetic parameters
1. gastric infusion hydroxytyrosol
Software is moved using 3P97 medicines pharmacokinetic parameters are calculated using one compartment model to blood concentration result.Gavage hydroxytyrosol
Pharmacokinetic parameters the results are shown in Table 5.Hydroxytyrosol mean blood plasma concentration-time graph is shown in Figure 14.
Pharmacokinetic parameters (n=5) after the gastric infusion HT of table 5 in blood plasma
2. gastric infusion hydroxytyrosol NO derivatives
Software is moved using 3P97 medicines pharmacokinetic parameters are calculated using two compartment model to HT-NO blood concentrations result, to administration
Hydroxytyrosol blood concentration result after HT-NO calculates pharmacokinetic parameters using one compartment model.Its pharmacokinetics is joined after gavage HT-NO
Number the results are shown in Table 6, and mean blood plasma concentration-time graph is shown in Figure 15；Hydroxytyrosol pharmacokinetic parameters the results are shown in Table after gavage HT-NO
7, mean blood plasma concentration-time graph is shown in Figure 16；
Pharmacokinetic parameters (n=5) of the HT-NO in blood plasma after the gastric infusion HT-NO of table 6
Pharmacokinetic parameters (n=5) of the HT in blood plasma after the gastric infusion HT-NO of table 7
1.4 experiment conclusion
(1) method is easy, sensitive, reproducible, can be applied to determine in blood plasma hydroxytyrosol and HT-NO content and should
For the pharmacokinetic study of gavage hydroxytyrosol and HT-NO in rat body.
(2) HT-NO tachymetabolism can discharge hydroxytyrosol and play its pharmacology and physiological action in vivo.
Embodiment 15
The NO releases of hydroxytyrosol and HT-NO in rat body are evaluated
Materials and methods
1.1 trial drug
Embodiment 2 prepares gained compound
Experiment material
Reagent：Bioengineering Research Institute, article No. are built up in NO kits, Nanjing：A012.
Key instrument：ELIASA；WH-2 micro-whirlpool vortex mixers, Shanghai Hu Xi analytical instrument company；GENIUS 16K-R
Low-temperature and high-speed centrifuge, Changsha Xin Ao instrument and meters Co., Ltd；DK98-1 thermostat water baths, the limited public affairs of Tianjin Stettlen instrument
Department.
Experimental animal：Male SD rat, weight 250-300g, by Xi'an Communications University, animal experimental center is provided.
1.2 experimental method
Rat is distinguished after gavage hydroxytyrosol and HT-NO, takes rat artery blood to centrifuge according to the time point of pharmacokinetic studies
Blood plasma is obtained, the dynamic level of active NO in 24h in rat plasma is evaluated.Determined using Nitrate reductase in rat plasma
NO2-、NO3-, (kit builds up Bioengineering Research Institute by Nanjing and provided) is operated by kit specification, enzyme mark is utilized
Instrument is measured, and measure wavelength is 550nm.
1.3 experimental result
Difference gastric infusion 50mgkg-1Hydroxytyrosol and 65mgkg-1HT-NO after, NO in 24h in rat serum
Dynamic level see Figure 17.
1.4 experiment conclusion
(1) HT-NO can discharge NO rapidly in rat body, reach higher level.
(2) after administration hydroxytyrosol in rat body NO levels to be relatively administered physiological saline high, this is due to that hydroxytyrosol can be with
Increase NO synthesis.
Claims (6)
1. a kind of hydroxytyrosol NO donor derivatives, it is characterised in that its chemical structural formula is：
R isWherein n=2.
2. a kind of preparation method of hydroxytyrosol NO donor derivatives described in claim 1, it is characterised in that：
Synthesis step is as follows：
1) 3,4- dihydroxyphenyl ethanols are substituted reaction and 4- (2- chloroethyls) -1,2- benzenediols are made；
2) 5- (2- chloroethyls) -2,2- diphenyl benzenes are made with dichloro diphenyl methane reaction in 4- (2- chloroethyls) -1,2- benzenediols
And [1,3] dioxole；
3) with silver nitrate nucleophilic substitution system occurs for 5- (2- chloroethyls) -2,2- diphenyl benzo [1,3] dioxole
Obtain 5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole；
4) 4- (2- are made by acidic hydrolysis in 5- (2- nitre trimethylammonium) -2,2- diphenyl benzo [1,3] dioxole
Nitre trimethylammonium) -1,2- benzenediols.
3. a kind of application of hydroxytyrosol NO donor derivatives described in claim 1, it is characterised in that：Preparing prevention and controlling
Treat the application in the medicine of diabetic vascular complications.
4. a kind of application of hydroxytyrosol NO donor derivatives according to claim 3, it is characterised in that：Preparing blood vessel
Application in dilator.
5. a kind of application of hydroxytyrosol NO donor derivatives according to claim 3, it is characterised in that：It is used in preparation
Application in blood vessel endothelium protection medicine.
6. a kind of application of hydroxytyrosol NO donor derivatives according to claim 3, it is characterised in that：Preparing, blood drops
Application in sugared medicine.
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Citations (2)
| 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 |
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| CN103687843A (en) * | 2011-10-24 | 2014-03-26 | 尼科斯股份有限公司 | Quinone based nitric oxide donating compounds |
Non-Patent Citations (5)
| Title |
|---|
| Isolation, synthesis, and neurite outgrowth-promoting activity of illicinin A from the flowers of Illicium anisatum;Shigeki Takaoka等;《Tetrahedron》;20090811;第65卷;第8356页Scheme 3 * |
| NO-Donor Phenols: A New Class of Products Endowed with Antioxidant and Vasodilator Properties;Donatella Boschi等;《J.Med.Chem.》;20060422;第49卷(第10期);第2886页摘要，第2887页Chart 1、Scheme 1 * |
| Structure–Antioxidant Activity Relationships in a Series of NO-Donor Phenols;Paolo Tosco等;《ChemMedChem》;20081231;第3卷;第1443-1448页 * |
| Synthesis and Antihypertensive Activity of Some Ester Progenitors of Methyldopa;Walfred S.Saari等;《Journal of Medicinal Chemistry》;19781231;第21卷(第8期);第747页左栏第1段 * |
| 羟基酪醇作为线粒体营养素的调控机制;罗成，刘健康;《生命科学》;20121031;第24卷(第10期);第1117页右栏倒数第2段 * |
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