阅读说明：本技术 一类3′-氨烷氧基-木犀草素衍生物及其制备方法和应用 (A kind of 3 '-aminoalkoxides-luteolin derivative and its preparation method and application ) 是由 杨为民 李鲜 翁稚颖 刘伟军 张秀娟 陈晨 肖创 白春昀 郑昌博 于 2018-12-21 设计创作，主要内容包括：本发明涉及一类3′-氨烷氧基-木犀草素衍生物的制备和应用。在惰性气体保护下,以木犀草素、具有苄基的活性基团物质为原料,在碱性条件下反应,得到二苄基取代物,再与具有氨基的活性基团,在碱性条件下反应,将反应产物在金属催化剂条件下,脱去苄基转化为3’-氨烷氧基-木犀草素衍生物。本发明化合物可进一步成盐。本发明所述化合物具有拮抗糖尿病大鼠血管损伤,拮抗大鼠心肌缺血损伤和脑缺血损伤等多靶点作用,通过保护血管发挥有关药理药效作用,可用于制备防治糖尿病血管并发症及心脑血管疾病的药物或保健品等。(The present invention relates to the preparations and application of a kind of 3 '-aminoalkoxides-luteolin derivative.Under inert gas protection; using luteolin, the active group substance with benzyl as raw material; it reacts under alkaline condition; obtain dibenzyl substituent; again with the active group with amino; it reacts under alkaline condition, by reaction product under metal catalysis conditions, sloughs benzyl and be converted into 3'- aminoalkoxide-luteolin derivative.The compounds of this invention can be further at salt.Compound of the present invention has antagonism diabetes rat injury of blood vessel; the multiple target effects such as the damage of antagonism myocardial ischemia in rats and cerebral ischemia; related pharmacological effect effect is played by protection blood vessel, can be used for preparing drug or health care product of prevention and treatment diabetic vascular complications and cardiovascular and cerebrovascular disease etc..)

1. one kind 3'- aminoalkoxide-luteolin derivative and its pharmaceutically can at salt, it is characterised in that have following formula knot Structure:

Wherein, R representative-(CH₂)nNR₁R₂、Wherein R₁And R₂It is identical or different, represent H or The alkyl of the linear chain or branched chain of C1-C6, n represent 1~6 integer, R₃Or R₄Represent the alkyl of the linear chain or branched chain of H or C1-C6.

2. 3'- aminoalkoxide-luteolin derivative according to claim 1 and its pharmaceutically can at salt, feature It is the R₁And R₂It is identical or different, H, methyl or ethyl are represented, n represents 1~4 integer, R₃Or R₄Represent H, methyl or second Base.

3. 3'- aminoalkoxide-luteolin derivative according to claim 2 and its pharmaceutically can at salt, feature It is the R representative-(CH₂)₂N(CH₃)₂、–(CH₂)₂NHCH₃、–(CH₂)₂NH₂、–(CH₂)₃N(CH₃)₂、

4. 3'- aminoalkoxide-luteolin derivative according to claim 1-3 and its pharmaceutically can at Salt, it is characterised in that it is described pharmaceutically can at salt be inorganic acid salt or acylate.

5. 3'- aminoalkoxide-luteolin derivative according to claim 4 and its pharmaceutically can at salt, feature It is that the inorganic acid salt or acylate are selected from sulfate, phosphate, hydrochloride, hydrobromate, acetate, oxalates, lemon Lemon hydrochlorate, succinate, gluconate, tartrate, tosilate, benzene sulfonate, mesylate, benzoate, Lactate or maleate.

6. according to claim 1 any one of -5 3'- aminoalkoxide-luteolin derivatives and its pharmaceutically can at salt Preparation method, it is characterised in that using luteolin, the active material with benzyl as raw material, react, obtain under alkaline condition Luteolin dibenzyl substituent, then the active group with amino is introduced in the 3' position hydroxyl of luteolin dibenzyl based compound Group, is prepared luteolin derivative after sloughing protecting group benzyl.

7. preparation method according to claim 6, it is characterised in that include the following steps:

(1) under inert gas protection, luteolin alkaline solution is prepared, being added, there is the active material of benzyl to react；

(2) being added into the reaction product of step (1), there are substance, triphenylphosphine and the anhydrous non-alcohols of amino active group to have Diethyl azodiformate or diisopropyl azodiformate reaction is added in solvent, stirring；

(3) step (2) reaction product is dissolved with methanol, H₂Reaction system is replaced, metallic catalyst is added, sloughs protecting group benzyl Base obtains 3'- aminoalkoxide-luteolin derivative of the present invention.

8. preparation method according to claim 7, it is characterised in that step (1) inert gas is selected from nitrogen or argon gas One or more of, the active material of the benzyl is selected from benzyl chloride, bromobenzyl, one or more of iodine benzyl, and alkali used is selected from Diisopropylethylamine, triethylamine, pyridine or DBU；

Step (2) substance with amino active group is selected from HO (CH₂)nNR₁R₂、 Wherein R₁And R₂It is identical or different, represent the alkyl or tertiary butyloxycarbonyl of the linear chain or branched chain of H or C1-C6 Base, n represent 1~6 integer, R₃Or R₄Represent the alkyl or tertbutyloxycarbonyl of the linear chain or branched chain of H or C1-C6；

Step (3) metallic catalyst is selected from Pd/C, Au-Pd/TiO₂Or Ag/ZnO.

9. preparation method according to claim 7 or 8, it is characterised in that step (1)-(3) further include producing to reaction The purification procedures of object, or including step (4) by product further with acid at salt.

10. according to claim 1 any one of -5 luteolin derivatives and its pharmaceutically can at salt in preparation prevention and Treat the application in ischemia injury drug or health care product.

11. application according to claim 10, it is characterised in that the drug or health care product are to prevent and treat diabetes Vascular complication and ischemic angiocardiopathy and cerebrovascular disease drug or health care product.

12. application according to claim 11, it is characterised in that the diabetic complication includes diabetic nephropathy, glycosuria Sick retinopathy, diabetes, the ischemic angiocardiopathy and cerebrovascular disease include treating myocardial ischemia damage and cerebral ischemia.

Technical field

The invention belongs to field of medicinal chemistry, and in particular to a kind of 3 '-aminoalkoxides-luteolin derivative and its preparation Methods and applications.

Background technique

Diabetes (Diabetes Mellitus, DM) are a kind of metabolic diseases characterized by hyperglycemia, in the world glycosuria Patient's number increases year by year, has 4.15 hundred million at present；Yellow is susceptible high-incidence, and China has become in the world that diabetic is most More country'ies (illness rate is up to 11.6%), 90% is diabetes B in diabetic.Long term hyperglycemia makes big blood vessel, micro- Vascular injury, and seriously injure the heart, brain, kidney, peripheral nerve, eyes, foot etc., leading to diabetes is the most disease of complication (up to more than 100 kinds).It is counted according to American Diabetes Association, diabetic's illness appears above the probability of complication for 3,5,10 years Respectively 46,61,98%.Diabetic vascular complications (Diabetes mellitus vascular complications, DMVC) be DM most serious complication, be diabetes lethal the main reason for disabling, it is concurrent that 80% diabetic dies of this Disease.DMVC mainly includes macrovascular complications (coronary heart disease, headstroke and atherosclerosis etc.) and microvascular complication (glycosuria Sick nephrosis, retinopathy, diabetes and peripheral neuropathy etc.).DMVC is definite, and pathogenesis is imperfectly understood, hyperglycemia The primary power for causing vascular lesion, mainly with glycometabolism, disorders of lipid metabolism and hypertension etc. in diabetes it is a variety of it is comprehensive because Element, it is related that induction of vascular damages (VD).VD includes blood vessel endothelium, vascular smooth injury of muscle, respectively with vascular endothelial cell, smooth Myocyte is related, leads to the functional disturbances such as vasomotoricity, endothelium secretion, barrier.Its pathology molecular mechanism relates generally to oxygen Change, inflammation stress reaction etc..

It clinically there is no the protective agents specifically for DMVC at present, it is main using adjusting blood glucose, blood lipid, blood pressure, and The drugs such as anti-oxidant, anti-inflammatory and antithrombotic are prevented and treated, and specifically include that Roger's column that are hypoglycemic and increasing insulin sensitivity Ketone, melbine and sulfonylurea drugs；Adjust the statins (Lovastatin etc.) of blood lipid；The angiotensins of blood pressure lowering turns Change enzyme inhibitor (ACEI) drug (captopril etc.) and calcium antagonist depressor (nifedipine)；Inhibit thrombus, improves blood The aspirin etc. of fluid rheology.In addition, that is researching and developing has anti-inflammatory drug such as JAK-STAT inhibitor, Chinese medicine blood-activating stasis-removing kind Drug, flavone compound etc..It is external that clinical test was once carried out using simple antioxidant, but result is unsatisfactory.DMVC Belong to complex disease, using the simple method by blood lipid, blood pressure, the drug of single target spot intervention can not be to this kind of disease It is effectively controlled and is treated, it is often more important that these drugs have more side effect, are more toxic, and are unfavorable for long-term Use.Therefore, single target drug is difficult to effectively treat diabetic vascular complications caused by non-single-gene or non-single target spot And control, therefore design and research and development multiple target point drug may be alleviation and treat one of diabetic vascular complications and new have efficacious prescriptions Method.

Ischemic angiocardiopathy and cerebrovascular disease (ICCVD) is the principal pathogenetic type of cardiovascular and cerebrovascular disease, ischemia-reperfusion (IR) Damage is the main pathologic process of ischemic angiocardiopathy and cerebrovascular disease, and IR leads to injury of blood vessel, inflammatory reaction, oxidative stress, thrombus Formed etc..From pathogenesis, ICCVD belongs to the more complex complex disease of pathogenesis, pathomechanism and its complexity Pathology network is related, therefore clinically prevention and treatment ICCVD will be directed to its pathology network mechanism, using comprehensive therapeutic intervention method, Alleviate ischemic, anti-inflammatory treatment, anti-oxidation stress, antithrombus formation, the measures such as inducing ischemia pre-adaptation progress including expansion blood vessel Prevention and treatment, therefore multiple target point intervention may be the effective ways of clinical prevention ICCVD.

Luteolin (Luteolin, LTD) is the natural flavone compound being present in a variety of medicinal plants and vegetables, is changed Scientific name is known as luteolin, has the pharmacology such as anti-oxidant, antitumor, anti-inflammatory, hypoglycemic, antagonizing vessel complication Effect.However the poorly water-soluble of luteolin, cause its druggability bad, bioavilability is low, limited efficacy.Improve reseda The water solubility and drug effect of plain compound, research and development activity is strong, toxic side effect is low, structure novel luteolin derivative is reseda The research direction of plain compound structure transformation.

Summary of the invention

It is derivative to provide a kind of 3 '-aminoalkoxides-luteolin in view of the above-mentioned problems, using chemical synthesis by the present invention Object plays antagonism diabetic vascular injury, antagonism treating myocardial ischemia damage and cerebral ischemia effect by protecting blood vessel.

Technical scheme is as follows:

A kind of 3 '-aminoalkoxides-luteolin derivative and its pharmaceutically can at salt, there is formula:

Wherein, R representative-(CH₂)nNR₁R₂、Wherein R₁And R₂It is identical or different, it represents The alkyl of the linear chain or branched chain of H or C1-C6, n represent 1~6 integer, R₃Or R₄Represent the alkane of the linear chain or branched chain of H or C1-C6 Base.

Preferably, the R₁And R₂It is identical or different, H, methyl or ethyl are represented, n represents 1~4 integer, R₃Or R₄Generation Table H, methyl or ethyl.

It is furthermore preferred that the R representative-(CH₂)₂N(CH₃)₂、–(CH₂)₂NHCH₃、–(CH₂)₂NH₂、–(CH₂)₃N(CH₃)₂、Specific structure is as follows:

3 '-aminoalkoxides-luteolin derivative of the present invention further includes the inorganic acid salt of the derivative or is had Machine hydrochlorate.The inorganic acid salt or acylate be selected from sulfate, phosphate, hydrochloride, hydrobromate, acetate, oxalates, Citrate, succinate, gluconate, tartrate, tosilate, benzene sulfonate, mesylate, benzoic acid Salt, lactate or maleate.

Another object of the present invention is to provide one kind 3 '-aminoalkoxides-luteolin derivative of the present invention and its medicine On can at the preparation method of salt reacted under alkaline condition using luteolin, the active material with benzyl as raw material, Luteolin dibenzyl substituent is obtained, then introduces amino active group in 3 ' position hydroxyls of luteolin dibenzyl based compound, Luteolin derivative is prepared after sloughing protecting group benzyl.

Synthetic route is schematically as follows:

A preferred preparation method of the invention, includes the following steps:

(1) under inert gas protection, luteolin alkaline solution is prepared, being added, there is the active material of benzyl to react；

(2) substance, triphenylphosphine and the anhydrous non-alcohol with oxygroup active group are added into the reaction product of step (1) Diethyl azodiformate or diisopropyl azodiformate reaction is added in class organic solvent, stirring；

(3) step (2) reaction product is dissolved with methanol, H₂Reaction system is replaced, metallic catalyst is added, sloughs protection Base benzyl obtains 3 '-aminoalkoxides-luteolin derivative of the present invention.

Above-mentioned preparation method, step (1) inert gas is selected from one or more of nitrogen or argon gas, described to have The active material of benzyl is selected from diisopropylethylamine, triethylamine selected from benzyl chloride, bromobenzyl, one or more of iodine benzyl, alkali used, Pyridine or DBU, solvent are conventional organic solvent, and a specific embodiment is solvent using DMF.

Step (2) substance with amino active group is selected from HO (CH₂)nNR₁R₂、 Wherein R₁And R₂It is identical or different, represent the alkyl or tertiary butyloxycarbonyl of the linear chain or branched chain of H or C1-C6 Base, n represent 1~6 integer, R₃Or R₄Represent the alkyl or tertbutyloxycarbonyl of the linear chain or branched chain of H or C1-C6.Preferably, institute State R₁And R₂It is identical or different, H, methyl or ethyl are represented, n represents 1~4 integer, R₃Or R₄Represent H, methyl or ethyl.More Preferably, 2-dimethylaminoethanol, N- tertbutyloxycarbonyl-N- methyl amino ethanol, N- (tertbutyloxycarbonyl) ethanol amine, 3- are selected from Dimethylamino -1- propyl alcohol, N- tertbutyloxycarbonyl -4- hydroxy piperidine or 1- tertbutyloxycarbonyl -3- hydroxyl pyrrolidine.

Preferred steps (2) reaction temperature is 15 DEG C -20 DEG C, and the reaction time is -48h for 24 hours.

Step (3) metallic catalyst is selected from Pd/C, Au-Pd/TiO₂Or Ag/ZnO.

Above-mentioned preparation method, step (1)-(3) further include the purification procedures to reaction product, or including step Suddenly (4) are by product further with acid at salt.

A specific preparation method of the invention, includes the following steps:

(1) under inert gas protection, luteolin alkaline solution is prepared, 10min is stirred, it is living to instill benzyl under ice-water bath Property substance, 15-20 DEG C stir 24 hours, TLC detection.

(2) mixed solution in step (1) is concentrated, 100mL ethyl acetate and 200mL H is added₂O stirs 30min, Filtering, organic phase are concentrated to give crude product.Crude product passes through silica gel chromatography.

(3) gained purified fraction in step (2) is placed in reflux condensate device, being added has amino active group object Matter, triphenylphosphine and anhydrous THF stir 10min, DEAD are instilled under ice-water bath, control 15 DEG C -20 DEG C of reaction temperature, LC-MS inspection It surveys, reaction solution concentration, silica gel chromatography.

(4) enriched fraction in step (3) is dissolved with methanol, H2 replaces reaction system, metallic catalyst is added, using washing It washs, filter, extracting, rotating one such or a variety of methods acquisition products, then with 1N hydrochloric acid at salt.

One preferred purification schemes of step (2) are as follows: LTD-Bn2 crude product is through silica gel chromatography, eluant, eluent: EA/ PE=1/5-1/3.LTD2 crude product is through silica gel chromatography, eluant, eluent: EA/PE=1/3-1/1.

Another object of the present invention be to provide 3 '-aminoalkoxides-luteolin derivative and its pharmaceutically can at salt exist Preparation prevents and treats the application in ischemia injury drug.Preferably, the drug is to prevent and treat diabetic vascular simultaneously Send out disease and ischemic angiocardiopathy and cerebrovascular disease drug, the diabetic complication include diabetic nephropathy, diabetic retinopathy, Diabetes, diabetes complicated injury of blood vessel, the ischemic angiocardiopathy and cerebrovascular disease include treating myocardial ischemia damage and cerebral ischemia damage Wound, such as coronary heart disease, headstroke.

The invention has the advantages that

(1) present invention uses chemical synthesis, and it is derivative to obtain a kind of 3 '-aminoalkoxide of multiple target point compound-luteolin Object and its pharmaceutically can at salt, compared to luteolin have good dissolubility and bioavilability, have higher guarantor Protect vasoactive.

(2) the compounds of this invention has antagonism diabetic vascular injury (but not influencing blood glucose), antagonism treating myocardial ischemia damage With cerebral ischemia multiple pharmacological effect, mechanism of action has the characteristics that multiple target point, is suitable for cardiovascular and cerebrovascular disease and diabetes The prevention and treatment of caused complication.

(3) current clinically main ICCVD protective agents are mostly based on single target drug, such as calcium ion antagonist Deng, and the present invention belongs to from multiple target points, the angle researching and designing correlation new drug candidate compound such as protection blood vessel, anti-inflammatory, anti-oxidant The new drug development direction of innovation.

(4) luteolin water solubility is low, causes to prepare that ease of formulation is big, and bioavilability is low, researching and designing one of the present invention Series derivates are intended to improve drug water solubility, while improving or keeping its vasoactive.A series of spread out what the present invention studied In biology, LTD3 water solubility highest but almost without vasoactive (being shown in Table 1, Figure 13), LTD10 vasoactive is best but water-soluble Property bad (LTD close), therefore preferably LTD2 is as main study subject.

Detailed description of the invention

The LCMS that Fig. 1 is compound L TD2 of the present invention schemes.

Fig. 2 is compound L TD2's of the present invention¹H-NMR figure.

The LCMS that Fig. 3 is compound L TD7 of the present invention schemes.

Fig. 4 is compound L TD7's of the present invention¹H-NMR figure.

The LCMS that Fig. 5 is compound L TD8 of the present invention schemes.

Fig. 6 is compound L TD8's of the present invention¹H-NMR figure.

The LCMS that Fig. 7 is compound L TD9 of the present invention schemes.

Fig. 8 is compound L TD9's of the present invention¹H-NMR figure.

The LCMS that Fig. 9 is compound L TD10 of the present invention schemes.

Figure 10 is compound L TD10's of the present invention¹H-NMR figure.

The LCMS that Figure 11 is compound L TD11 of the present invention schemes.

Figure 12 is compound L TD11's of the present invention¹H-NMR figure.

Figure 13 is the dose-effect of compound L TD2 of the present invention and other derivative diastole isolated rat aorta pectoralis blood vessels Curve graph.

Figure 14 is the laser speckle instrument detection that compound L TD2/LTD of the present invention increases ZDF rat pedal skin blood flow Figure.

Figure 15 is the statistical chart that compound L TD2/LTD of the present invention increases ZDF rat foot blood flow.(note: with ZDF Group compares, * P < 0.05, * * P < 0.01.).

Figure 16 is that compound L TD2/LTD incubated in vitro of the present invention improves ZDF isolated rat coronary vasodilation Reaction.(note: compared with ZDF group, * * P < 0.01, * * * P < 0.001.).

Figure 17 is that compound L TD2/LTD gastric infusion of the present invention enhances ZDF isolated rat coronary vasodilation Reaction.(note: compared with ZDF group, * P < 0.05, * * * P < 0.001.).

Figure 18 is influence (A1- of the compound L TD2 gastric infusion of the present invention to ZDF renal tissues of rats pathomorphism A3:fa/+ rat normal kidney tissue structure, B1-B3:ZDF renal tissues of rats structure, C1-C3:LTD2 is to ZDF renal tissues of rats The influence of structure).

Figure 19 is shadow of the compound L TD2 gastric infusion ZDF rat of the present invention to aorta pectoralis Pathomorphologic It rings.

Figure 20 is the representative section figure of different disposal group ischemic myocardium.

Figure 21 is protective effect of the compound L TD2 of the present invention to rat myocardial ischemia and reperfusion.(note: with model group Compare,^**P<0.01,^***P<0.001.)。

Figure 22 is the representative section figure of different disposal group ischemic tissue of brain.

Figure 23 corrects ischemic percent by volume.(note: compared with model group (Model),^***P<0.001.)。

Figure 24 ischemic areas percentage.(note: compared with model group (Model),^***P<0.001.)。

Figure 25 Neurological deficits.(note: compared with model group (Model),^**P<0.01,^***P<0.001.)。

Specific embodiment

Illustrate specific steps of the invention by the following examples, but is not limited by the example.

Term as used in the present invention generally there are those of ordinary skill in the art usually to manage unless otherwise indicated The meaning of solution.The present invention is described in further detail combined with specific embodiments below and referring to data.It should be understood that the embodiment is In order to demonstrate the invention, it rather than limits the scope of the invention in any way.

In the examples below, the various processes and method being not described in detail are conventional methods as known in the art.

The present invention is further described combined with specific embodiments below.