CN109956952B

CN109956952B - Alpha-toosedarin derivative and preparation method and application thereof

Info

Publication number: CN109956952B
Application number: CN201711336313.4A
Authority: CN
Inventors: 占扎君; 单伟光; 陈艳; 舒仁欢; 王建伟; 马列峰
Original assignee: Zhejiang University of Technology ZJUT
Current assignee: Shanghai ruling biomedical Co.,Ltd.
Priority date: 2017-12-14
Filing date: 2017-12-14
Publication date: 2020-11-13
Anticipated expiration: 2037-12-14
Also published as: CN109956952A; WO2019114501A1

Abstract

The invention discloses an alpha-toosendanin derivative with a general formula I structure, a preparation method thereof and application thereof in preparing medicaments for treating brain injury diseases caused by ischemia, such as vascular dementia and cerebral infarction; the vascular dementia is characterized by vascular lesion, learning and memory disorder caused after cerebral ischemia or cell loss in a dentate gyrus region of hippocampus.

Description

Alpha-toosedarin derivative and preparation method and application thereof

Technical Field

The invention relates to an alpha-toosendanin derivative, a preparation method of the alpha-toosendanin derivative and an application of the alpha-toosendanin derivative in preparing a medicine for treating ischemic brain injury, belonging to the technical field of medicines.

Background

Brain diseases caused by vascular diseases become a serious health hazard, and the problem is more prominent particularly with the increasing population of the elderly in China. The cerebral infarction refers to ischemic necrosis or brain softening of local brain tissue caused by cerebral blood supply disorder, ischemia and anoxia, the common clinical types of cerebral infarction include cerebral thrombosis, lacunar infarction, cerebral embolism and the like, and the cerebral infarction accounts for 80% of all cerebral apoplexy. Vascular Dementia (VD) is a dementia syndrome caused by brain tissue damage caused by cerebrovascular factors, the incidence of the vascular dementia accounts for about 20 percent of the causes of senile cognitive impairment, the vascular dementia is a cognitive disorder disease with the clinical incidence ranked second after Alzheimer's disease at present, and the life quality of patients is seriously influenced. The etiology of the vascular dementia is unknown so far, and scientific research shows that the risk factors causing the vascular dementia are numerous, and the pathogenic factors are complex, mainly including whole cerebral ischemia and ischemic stroke caused by cardiac arrest, and also including chronic disease factors such as hypertension, hyperlipidemia, diabetes and the like. The pathological mechanism of cognitive disorder caused by vascular dementia may be similar to that of common cognitive disorder and cognitive disorder caused by other dementia, but has specificity. In general, the pathological mechanisms of vascular dementia include vascular lesions, white matter damage, genetic influence and the like, wherein the vascular lesions are the basis of the pathogenesis of the vascular dementia.

The medicines for treating cerebral infarction mainly comprise edaravone and butylphthalide, and the rest are some medicines for preventing, such as thrombolytic medicines. Edaravone is not good in therapeutic effect and has a large side effect, and its main effect is through antioxidation. Butylphthalide has good effect on treating cerebral infarction, has the main effect of resisting inflammation, but has no obvious treatment effect on vascular dementia. At present, the clinical treatment of vascular dementia mainly takes preventive measures such as controlling risk factors and the like, therapeutic drugs (such as cholinesterase inhibitors) only relieve symptoms, and no targeted drugs for the vascular dementia exist in the drugs on the market, so that a new preventive and therapeutic drug is urgently needed to be found to deal with the vascular dementia.

The compound of the invention not only can treat cerebral infarction, but also has good treatment effect on vascular dementia. Mechanism research the compound of the invention has multiple functions of anti-inflammation, antioxidation, nerve protection, nerve differentiation and the like, and has the advantages of edaravone and butylphthalide. Based on the fact that the etiology of the cerebral infarction and vascular dementia is greatly related to vascular lesions, rather than a single factor causing nerve cell damage, a better treatment effect is difficult to achieve by aiming at the single factor, for example, some compounds have a neuroprotective effect or antioxidation, but do not have the effect of treating the cerebral infarction or vascular dementia. The compound has multiple effects, and a large number of animal experimental model evaluations find that the compound has good treatment effects on the cerebral infarction and the vascular dementia, so that the compound can be clinically used for treating the cerebral infarction and the vascular dementia. In addition, the compound has good safety, thereby having great medicinal prospect.

Disclosure of Invention

In order to achieve the purpose, the invention provides an alpha-toosendanin derivative, a preparation method of the alpha-toosendanin derivative and application of the alpha-toosendanin derivative in preparing a medicament for treating ischemic brain injury.

The alpha-azadirachtin derivative is a compound shown as a general formula I.

Wherein R is₁Is H, C₁-C₆A linear or branched alkyl group; r₂Is C₅-C₆A cyclic hydrocarbon group of₁-C₈The substituted phenyl group has a substituent of methyl, ethyl, chlorine, bromine, fluorine, methoxy, amino or nitro; or R₁、R₂Form a ring with NThe ring is a six-membered ring heterocycle containing N or containing N and O at the same time.

Further, said R₁Preferably methyl or ethyl.

Further, said R₂Preferably phenyl or methylphenyl.

Further, said R₁、R₂And N form a ring, and the ring is morpholinyl, piperazinyl, N-substituted piperazinyl or piperidinyl.

Furthermore, the alpha-azadirachtin derivative shown in the formula I is one of the following compounds:

further, the preparation method of the alpha-toosendanin derivative shown in the formula I is characterized by comprising the following steps: the alpha-toosendanin derivative is prepared by the following method:

(1) dissolving triphosgene in dichloromethane to prepare a dichloromethane solution of triphosgene with the concentration of 0.1-0.5 mol/L, dropwise adding a dichloromethane solution of a nitrogen-containing compound shown as a formula III with the concentration of 0.1-0.5 mol/L and triethylamine in the triphosgene solution in ice bath, and stirring at room temperature for 4-6 hours to obtain a reaction mixed solution A; the mass ratio of the triphosgene to the nitrogen-containing compound and the triethylamine shown in the formula III is 1: 0.67-1.5: 1.33-3;

(2) dropwise adding a dichloromethane solution of a compound shown in a formula II with the concentration of 0.05-0.25 mol/L into the reaction mixed solution A in an ice bath, continuously stirring for 4-6 h, and after the reaction is finished, carrying out post-treatment on the obtained reaction mixed solution B to obtain an alpha-toosendanin derivative shown in the formula I; the mass ratio of the compound shown in the formula II to the triphosgene is 1: 2.0 to 3.0.

Wherein R is₁Is H, C₁-C₆Straight or branched alkyl, R₂Is C₅-C₆A cyclic hydrocarbon group of₁-C₈The substituted phenyl group has a substituent of methyl, ethyl, chlorine, bromine, fluorine, methoxy, amino or nitro; or R₁、R₂And N to form a ring, wherein the ring is a six-membered cyclic heterocyclic ring containing N or containing N and O simultaneously.

Further, the post-treatment method of the reaction mixed liquid B comprises the following steps: adding distilled water with the volume 3-6 times that of the reaction mixed solution B into the obtained reaction mixed solution B, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, drying by using anhydrous Na2SO4, performing suction filtration, concentrating to obtain a yellow oily crude product, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate which shows strong ultraviolet at 254nm and has polarity less than that of the corresponding compound shown in formula II, combining the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain the alpha-toosendanin derivative shown in formula I.

Further, the compound of formula II according to the present invention is prepared as follows:

dissolving alpha-azadirachtin in toluene to prepare a toluene solution with the concentration of 0.1-0.5 mol/L, adding 2-3 times of equivalent of 2, 3-dichloro-5, 6 dicyanobenzoquinone (DDQ) into the solution at room temperature, reacting at 40 ℃ for 5-8 hours after the solution is completely dissolved, detecting by a thin layer, distilling under reduced pressure after the reaction is finished to remove the toluene, separating and purifying by silica gel column chromatography, and using petroleum ether: the volume ratio of ethyl acetate is 10: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate of a compound which shows strong ultraviolet at 254nm and has a polarity less than that of alpha-toosendanin, combining the eluates, evaporating the eluate under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain a compound shown as a formula II; the alpha-azadirachtin: the mass ratio of DDQ was 1: 2.0 to 3.0.

Still further, the application of the alpha-toosendanin derivative in preparing the medicine for treating the brain injury diseases caused by ischemia is provided, wherein the brain injury diseases are preferably vascular dementia and cerebral infarction.

Furthermore, the vascular dementia is angiopathy, learning and memory disorder caused by cerebral ischemia or cell loss of dentate gyrus of hippocampus.

More specifically, the method adopts a Longa line-drawing method to prepare a rat focal cerebral ischemia-reperfusion injury model, so as to investigate the therapeutic effect of the alpha-toosendanin derivative on the cerebral ischemia-reperfusion injury model rat. The experimental result shows that the alpha-toosendanin derivative can obviously reduce the cerebral infarction area of a rat and reduce the water content of brain tissues, and the alpha-toosendanin derivative has a good treatment effect on cerebral infarction. The invention adopts MCAO wire-tying method to cause cerebral ischemia to simulate vascular dementia, evaluates the effect of the medicine on the space learning and memory ability of a rat with vascular dementia through Morris water maze experiment and open field experiment, observes the change of cell morphology in a hippocampal CA1 area through Niger dyeing, and explores the treatment effect of the alpha-toosendanin derivative on the vascular dementia. The experimental results show that the alpha-azadirachtin derivatives provided by the invention can shorten the incubation time of a platform for searching of rats induced by ischemia, the recovery of the searching behavior in an open field is obvious, and the damage of cells in a hippocampal CA1 area can be reversed. The alpha-toosendanin derivative has good treatment effect on vascular dementia.

Compared with the prior art, the invention has the beneficial effects that:

the alpha-azadirachtin derivative retains the active ingredients of the alpha-azadirachtin and reduces the toxicity of the compound. The alpha-toosendanin derivative has good treatment effect on vascular dementia, and the activity of the alpha-toosendanin derivative is stronger than that of a positive drug of polynaphthapine. The alpha-toosendanin derivative has a good treatment effect on cerebral infarction, the activity of the alpha-toosendanin derivative is obviously stronger than that of edaravone, and the alpha-toosendanin derivative is equivalent to butylphthalide; the improvement of the water content of the brain tissue is obviously better than that of edaravone and butylphthalide. This shows that the comprehensive treatment effect of the alpha-toosendanin derivative on the cerebral infarction is superior to that of the edaravone and butylphthalide which are first-line similar medicaments in the market. In addition, the alpha-toosendanin derivative has extremely high safety, experimental animals do not show any adverse symptoms under the condition of 2g/Kg intraperitoneal injection dose, and the median lethal dose of the parent compound alpha-toosendanin is 200 mg/Kg. Therefore, the alpha-toosedarin derivative has wide application prospect in preparing medicines for treating brain injury diseases caused by ischemia.

Drawings

FIG. 1 Effect of HZAM-1 administration according to the invention on the Morris water maze test in rats with vascular dementia.

FIG. 2 Effect of HZAM-1 administration according to the invention on the number of spanning lattices in the open field space exploration behavior of vascular dementia rats.

FIG. 3 Effect of HZAM-1 administration according to the invention on the number of forearm lifts in the open field space exploration behavior of rats with vascular dementia.

FIG. 4 Effect of HZAM-1 administration according to the invention on the median residence time in the open field space exploration behavior of rats with vascular dementia.

FIG. 5A graph of CA1 region of hippocampus in a Niger-stained sham operated group.

FIG. 6 is a graph of Hippocampus CA1 region of the Nicol staining model group.

FIG. 7 is a graph of CA1 region of Hippocampus of Nise staining HZAM-1-administered group.

Detailed Description

The invention is further illustrated with reference to the figures and examples, but the invention is not limited thereby within the scope of the described implementation.

Example 1: synthesis of alpha-azadirachtin derivative HZAM-1 (FIG. 1 for the scheme of the Synthesis)

Dissolving alpha-azadirachtin (40mg,0.1mmol) in toluene solution under stirring, adding 2, 3-dichloro-5, 6 dicyanobenzoquinone (DDQ,45mg,0.2mmol), reacting at 40 deg.C, detecting by TLC during the reaction, ending the reaction after 5 hr, distilling under reduced pressure to remove toluene, and separating and purifying by silica gel column chromatography to obtain yellow solid compound HZA with yield of 85%.

BTC (60mg,0.202mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, equivalent N-methylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an ice salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-1 with yield of 90%.

HZAM-1:ESI-MS m/z:540([M-H]^-,C₃₂H₃₁NO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.46-7.40(5H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.2Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.20(1H,s,H-14),4.12(1H,s,H-13),4.11(1H,s,H-13),3.46(3H,s,8-OCH₃),3.39(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.3,158.0,156.4,153.8,152.4,149.5,146.7,142.5,138.5,131.9,130.9,129.6,129.3,128.9,127.2,127.2,123.1,116.3,115.6,110.4,104.5,104.0,94.2,78.1,61.6,38.6,28.4,28.4,26.3,25.8,19.2.

Example 2: synthesis of alpha-azadirachtin derivative HZAM-2 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, equivalent N-ethylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an ice salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-2 with yield of 94%.

HZAM-2:ESI-MS m/z(％):556([M+H]⁺,C₃₃H₃₃NO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.45-7.36(5H,m,Ar-H),7.28(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.19(1H,s,H-14),4.11(1H,s,H-13),4.10(1H,s,H-13),3.86(3H,s,8-OCH₃),3.70(2H,m,N-CH₂),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.28(3H,m,N-CH₃)；¹³C-NMR:182.3,160.3,158.1,156.5,153.9,152.1,149.6,146.8,141.1,138.4,131.8,129.3,129.0,128.5,127.7,127.5,127.4,127.2,123.2,116.2,115.7,110.4,104.6,104.1,94.2,78.0,61.5,52.7,46.2,28.4,28.4,26.4,25.6,18.2.

Example 3: synthesis of alpha-azadirachtin derivative HZAM-3 (FIG. 1 shows the scheme of the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl o-toluidine, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an ice salt removing bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-3 with yield of 89%.

HZAM-3:ESI-MS m/z(％):554([M-H]^-,C₃₃H₃₃NO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.32-7.26(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.0Hz,H-3),5.17(1H,s,H-14),4.09(1H,s,H-13),4.08(1H,s,H-13),3.37(3H,s,8-OCH₃),3.35(3H,s,N-CH₃),2.40(3H,s,Ar-CH₃),1.81(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.7,152.5,149.6,146.6,141.0,138.4,135.6,131.8,131.1,128.3,127.4,127.2,126.4,123.1,123.0,116.1,115.6,110.1,104.5,104.0,94.1,78.1,61.3,38.8,28.4,28.4,26.3,25.8,18.2,17.4.

Example 4: synthesis of alpha-azadirachtin derivative HZAM-4 (FIG. 1 shows the scheme of the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, N-methyl m-methylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane which are equivalent are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in ice bath, after dropping, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-4 with a yield of 92%.

HZAM-4:ESI-MS m/z(％):556([M+H]⁺,C₃₃H₃₃NO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.33-7.19(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.20(1H,s,H-14),4.13(1H,s,H-13),4.12(1H,s,H-13),3.45(3H,s,8-OCH₃),3.37(3H,s,N-CH₃),2.40(3H,s,Ar-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.3,158.0,156.4,153.8,152.4,149.5,146.7,142.4,139.2,138.4,131.9,129.3,129.0,127.2,123.2,123.2,116.2,115.6,115.6,110.4,104.5,104.0,94.2,78.1,61.6,38.6,28.4,28.4,26.3,25.8,21.3,18.2.

Example 5: synthesis of alpha-azadirachtin derivative HZAM-5 (FIG. 1 shows the scheme of the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl-p-methylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-5 with a yield of 91%.

HZAM-5:ESI-MS m/z(％):556([M+H]⁺,C₃₃H₃₃NO₇)；¹H-NMR(500MHz):13.55(1H,s,5-OH),7.29-7.23(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.0Hz,H-3),5.20(1H,s,H-14),4.12(1H,s,H-13),4.11(1H,s,H-13),3.49(3H,s,8-OCH₃),3.41(3H,s,N-CH₃),2.38(3H,s,Ar-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.8,152.6,149.6,146.8,142.6,138.4,131.9,129.9,127.2,127.2,126.3,123.0,123.0,116.3,115.7,115.7,110.5,104.5,104.0,94.2,78.1,61.6,38.6,28.4,28.4,26.3,25.8,21.0,18.2.

Example 6: synthesis of alpha-Toosendanin derivative HZAM-6 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an N-methyl o-anisidine, triethylamine (0.25mmol) and 5mL of dichloromethane which are equivalent are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out under ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with mixed solvent, detecting with thin layer, and collecting eluate with strong ultraviolet at 254nm and polarity less than HZAAnd (3) eluent is combined, the eluent is evaporated under reduced pressure to remove an elution solvent, and a product obtained is dried to obtain the HZAM-6 with the yield of 88%.

HZAM-6:ESI-MS m/z(％):572([M+H]⁺,C₃₃H₃₃NO₈)；¹H-NMR(CDCl₃):13.57(1H,s,5-OH),7.34-7.30(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.0Hz,H-3),5.18(1H,s,H-14),4.10(1H,s,H-13),4.08(1H,s,H-13),3.90(3H,s,Ar-OCH₃),3.43(3H,s,8-OCH₃),3.32(3H,s,N-CH₃),1.81(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.3,160.2,158.0,156.4,155.1,153.2,153.0,149.8,146.6,138.2,131.8,131.0,129.2,128.8,127.1,123.0,120.8,115.9,115.6,111.8,110.1,104.5,104.0,94.1,78.0,61.2,55.6,37.7,28.4,28.4,26.3,25.8,18.1.

Example 7: synthesis of alpha-azadirachtin derivative HZAM-7 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl m-methoxyaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering, concentrating to obtain yellow oily crude product, and mixingThe material is prepared from petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-7 with a yield of 91%.

HZAM-7:ESI-MS m/z(％):572([M+H]⁺,C₃₃H₃₃NO₈)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.35-7.32(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.25(1H,s,H-12),5.59(1H,d,J＝10.0Hz,H-3),5.20(1H,s,H-14),4.13(1H,s,H-13),4.12(1H,s,H-13),3.84(3H,s,Ar-OCH₃),3.5(3H,s,8-OCH₃),3.34(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.3,160.2,158.0,156.4,153.8,152.4,149.5,146.7,143.6,138.5,131.9,129.9,127.2,127.2,123.1,123.1,116.3,115.6,112.6,110.5,104.5,104.0,94.2,78.1,61.2,55.5,38.6,28.4,28.4,26.4,25.8,18.2.

Example 8: synthesis of alpha-azadirachtin derivative HZAM-8 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl-p-anisidine, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out in ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers,washing with saturated NaCl solution, and washing with anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-8 with yield of 90%.

HZAM-8:ESI-MS m/z(％):570([M-H]^-,C₃₃H₃₃NO₈)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.32-6.94(4H,m,Ar-H),7.26(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.0Hz,H-3),5.18(1H,s,H-14),4.11(1H,s,H-13),4.10(1H,s,H-13),3.84(3H,s,Ar-OCH₃),3.5(3H,s,8-OCH₃),3.34(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.8,152.7,149.5,146.7,142.5,138.4,131.9,129.9,127.7,127.2,127.2,123.1,123.1,116.2,115.6,114.5,114.5 110.5,104.5,104.0,94.1,78.1,61.6,55.5,38.9,28.4,28.4,26.3,25.8,18.2.

Example 9: synthesis of alpha-azadirachtin derivative HZAM-9 (FIG. 1 shows the scheme of the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl o-ethylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. Slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, removing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-9 with yield of 89%.

HZAM-9:ESI-MS m/z(％):568([M+H]⁺,C₃₄H₃₅NO₇)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.36-7.28(4H,m,Ar-H),7.25(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.17(1H,s,H-14),4.08(2H,s,H-13),3.35(3H,s,8-OCH₃),3.34(3H,s,N-CH₃),2.75(2H,m,Ar-CH₂),1.80(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.33(3H,m,Ar-CH₃)；¹³C-NMR:182.2,160.3,158.0,156.4,153.8,152.7,149.6,146.6,141.3,140.6,138.4,131.9,129.4,128.5,127.6,127.2,127.1,123.1,116.1,115.7,110.4,104.6,104.0,94.2,78.1,61.3,38.4,28.4,28.4,26.4,25.8,23.7,18.2,14.5.

Example 10: synthesis of alpha-azadirachtin derivative HZAM-10 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, equivalent N-methyl m-ethylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, slowly dropwise adding is carried out under ice bath, after the dropwise adding is finished, an deicing salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single neck flask, a trace amount of DMAP, 5mL of dichloroMethane, magnetically stirred at room temperature until completely dissolved. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-10 with yield of 92%.

HZAM-10:ESI-MS m/z(％):568([M+H]⁺,C₃₄H₃₅NO₇)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.35-7.32(4H,m,Ar-H),7.26(1H,s,H-10),6.73(1H,d,J＝10.1Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.20(1H,s,H-14),4.11(2H,s,H-13),3.43(3H,s,8-OCH₃),3.43(3H,s,N-CH₃),2.70(2H,m,Ar-CH₂),1.82(3H,s,17-CH₃),1.67(3H,s,16-CH₃),1.47(6H,s,18,19-CH₃),1.26(3H,m,Ar-CH₃)；¹³C-NMR:182.3,160.3,158.0,156.4,153.8,152.6,149.6,145.6,142.5,138.5,131.9,129.1,127.2,127.2,123.0,123.0,116.2,115.6,115.6,110.4,104.5,104.0,94.2,78.1,61.6,38.4,30.9,28.4,28.4,26.4,25.8,18.2,15.4.

Example 11: synthesis of alpha-Azadirachta derivative HZAM-11 (FIG. 1 shows the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl-p-ethylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-11 with yield of 94%.

HZAM-11:ESI-MS m/z(％):568([M+H]⁺,C₃₄H₃₅NO₇)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.31-7.26(4H,m,Ar-H),7.25(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.19(1H,s,H-14),4.11(2H,s,H-13),3.47(3H,s,8-OCH₃),3.41(3H,s,N-CH₃),2.67(2H,m,Ar-CH₂),1.83(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.26(3H,m,Ar-CH₃)；¹³C-NMR:182.3,160.3,158.0,156.4,153.8,152.6,146.7,140.1,138.5,131.9,128.7,128.7,127.2,127.2,126.4,123.1,123.1,116.1,115.7,115.7,110.5,104.5,104.0,94.2,78.1,61.6,38.4,30.9,28.4,28.4,26.4,25.8,18.2,15.5.

Example 12: synthesis of alpha-azadirachtin derivative HZAM-12 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl o-chloroaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-12 with a yield of 88%.

HZAM-12:ESI-MS m/z(％):574([M-H]^-,C₃₂H₃₀ClNO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.52-7.34(4H,m,Ar-H),7.24(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.17(1H,s,H-14),4.10(1H,s,H-13),3.90(1H,s,H-13),3.45(3H,s,8-OCH₃),3.37(3H,s,N-CH₃),1.81(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.8,152.4,149.4,146.7,142.5,138.5,135.7,130.4,129.4,128.7,128.0,127.2,123.1,120.3,116.3,115.6,110.3,104.5,104.0,94.2,78.1,61.5,38.6,28.4,28.4,26.4,25.8,18.2.

Example 13: synthesis of alpha-azadirachtin derivative HZAM-13 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl-m-chloroaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-13 with a yield of 92%.

HZAM-13:ESI-MS m/z(％):574([M-H]^-,C₃₂H₃₀ClNO₇)；¹H-NMR(CDCl₃):13.55(1H,s,5-OH),7.32(2H,m,Ar-H),7.25(1H,s,H-10),6.98(2H,m,Ar-H),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.20(1H,s,H-14),4.12(1H,s,H-13),4.11(1H,s,H-13),3.57(3H,s,8-OCH₃),3.46(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.3,158.0,156.4,153.8,152.4,149.2,146.7,143.6,138.7,134.7,134.7,132.1,130.2,130.2,127.3,123.0,123.0,116.5,115.6,115.6,110.5,104.5,104.0,94.2,78.1,61.7,38.4,28.4,28.4,26.3,25.8,18.2.

Example 14: synthesis of alpha-Azadirachta derivative HZAM-14 (FIG. 1 shows the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, N-methyl parachloroaniline, triethylamine (0.25mmol) and 5mL of dichloromethane which are equivalent are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-14 with a yield of 92%.

HZAM-14:ESI-MS m/z(％):576([M+H]⁺,C₃₂H₃₀ClNO₇)；¹H-NMR(CDCl₃):13.52(1H,s,5-OH),7.42-7.34(4H,m,Ar-H),7.29(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.18(1H,s,H-14),4.10(1H,s,H-13),4.12(1H,s,H-13),3.44(3H,s,8-OCH₃),3.36(3H,s,N-CH₃),1.81(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.47(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.7,152.1,149.3,146.6,141.2,138.4,133.6,131.9,129.7,129.4,128.7,127.2,123.1,123.0,116.2,115.6,110.5,104.8,104.1,94.2,78.1,61.4,37.4,28.4,28.4,26.3,25.8,18.2.

Example 15: synthesis of alpha-Toosendanin derivative HZAM-15 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl o-bromoaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an ice salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-15 with yield of 90%.

HZAM-15:ESI-MS m/z(％):620([M+H]⁺,C₃₂H₃₀BrNO₇)；¹H-NMR(CDCl₃):13.54(1H,s,5-OH),7.48-7.40(4H,m,Ar-H),7.24(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.17(1H,s,H-14),4.10(1H,s,H-13),4.09(1H,s,H-13),3.45(3H,s,8-OCH₃),3.37(3H,s,N-CH₃),1.81(3H,s,17-CH₃),1.66(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.2,160.2,158.0,156.4,153.7,152.1,149.3,146.6,141.2,138.4,133.6,131.9,129.7,129.4,128.7,128.0,127.2,123.1,116.3,115.6,110.3,104.5,104.0,94.2,78.1,61.5,38.6,28.4,28.4,26.4,25.8,18.2.

Example 16: synthesis of alpha-azadirachtin derivative HZAM-16 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, N-methyl-m-bromoaniline, triethylamine (0.25mmol) and 5mL of dichloromethane which are equivalent are added into a 25mL constant pressure dropping funnel, the dropwise addition is slowly carried out under ice bath, after the dropwise addition is finished, an deicing salt bath is removed, and the stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-16 with a yield of 91%.

HZAM-16:ESI-MS m/z(％):620([M+H]⁺,C₃₂H₃₀BrNO₇)；¹H-NMR(CDCl₃):13.52(1H,s,5-OH),7.46-7.29(4H,m,Ar-H),7.24(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.20(1H,s,H-14),4.13(1H,s,H-13),4.12(1H,s,H-13),3.49(3H,s,8-OCH₃),3.45(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.47(6H,s,18,19-CH₃)；¹³C-NMR:182.3,160.3,158.0,156.4,153.8,152.2,149.2,146.7,143.7,138.6,132.0,130.5,127.2,127.2,123.0,123.0,122.5,116.3,115.6,115.6,110.5,104.6,104.0,94.2,78.1,61.7,38.4,28.4,28.4,26.4,25.8,18.2

Example 17: synthesis of alpha-azadirachtin derivative HZAM-17 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, equivalent N-methyl-p-bromoaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, slowly dropwise adding is carried out under ice bath, after the dropwise adding is finished, an deicing salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-17 with yield of 93%.

HZAM-17:ESI-MS m/z(％):618([M-H]^-,C₃₂H₃₀BrNO₇)；¹H-NMR(CDCl₃):13.52(1H,s,5-OH),7.57-7.29(4H,m,Ar-H),7.23(1H,s,H-10),6.74(1H,d,J＝10.0Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.19(1H,s,H-14),4.12(1H,s,H-13),4.11(1H,s,H-13),3.49(3H,s,8-OCH₃),3.43(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃)；¹³C-NMR:182.1,160.3,158.0,156.4,153.8,152.2,149.2,146.7,141.5,138.6,132.4,132.4,132.0,127.2,127.2,123.0,123.0,116.3,115.6,115.6,110.5,104.6,104.0,94.2,78.1,61.7,38.4,28.4,28.4,26.4,25.8,18.2.

Example 18: synthesis of alpha-azadirachtin derivative HZAM-18 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl-N-butylaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-18 with yield of 90%.

HZAM-18:ESI-MS m/z(％):596([M-H]^-,C₃₆H₃₉NO₇)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.31-7.28(4H,m,Ar-H),7.24(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.19(1H,s,H-14),4.11(2H,s,H-13),3.46(3H,s,8-OCH₃),3.42(3H,s,N-CH₃),2.64(2H,m,Ar-CH₂),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.60(2H,m,Ar-CH₂),1.48(6H,s,18,19-CH₃),1.37(2H,m,Ar-CH₂),1.27(2H,m,Ar-CH₂),0.94(3H,m,Ar-CH₃)；¹³C-NMR:182.3,160.3,158.0,156.4,153.8,152.6,149.6,146.7,140.1,138.4,131.9,129.2,129.2,127.2,127.2,126.4,123.1,123.1,116.1,115.7,110.4,104.5,104.0,94.2,78.1,61.6,38.6,35.2,33.5,28.4,28.4,26.3,25.8,22.3,18.2,13.9.

Example 19: synthesis of alpha-azadirachtin derivative HZAM-19 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, an equivalent amount of N-methyl o-ethoxyaniline, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, the mixture is slowly dropped in an ice bath, after the dropping is finished, an deicing salt bath is removed, and the mixture is stirred for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-19 with yield of 93%.

HZAM-19:ESI-MS m/z(％):584([M-H]^-,C₃₅H₃₄NO₈)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.31-7.28(4H,m,Ar-H),7.24(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.26(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.19(1H,s,H-14),4.12(1H,s,H-13),4.11(1H,s,H-13),4.06(2H,m,Ar-OCH₂),3.49(3H,s,8-OCH₃),3.44(3H,s,N-CH₃),1.83(3H,s,17-CH₃),1.68(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.26(3H,m,Ar-CH₃)；¹³C-NMR:182.3,160.2,158.0,156.4,153.8,152.4,149.5,146.7,143.8,138.4,131.9,129.8,127.2,127.2,123.0,123.0,116.3,115.6,113.2,110.4,104.5,104.0,94.2,78.1,61.6,58.4,38.6,28.4,28.4,26.3,25.8,18.2,14.7.

Example 20: synthesis of alpha-azadirachtin derivative HZAM-20 (FIG. 1 shows the scheme of the synthesis scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, 0.8 equivalent of pyrrolidine, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an ice salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-20 with a yield of 91%.

HZAM-20:ESI-MS m/z(％):506([M+H]⁺,C₂₉H₃₁NO₇)；¹H-NMR(CDCl₃)13.55(1H,s,5-OH),7.22(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.23(1H,s,H-14),4.16(1H,s,H-13),4.15(1H,s,H-13),3.79(3H,s,8-OCH₃),3.52(2H,m,Py-CH₂),3.44(2H,m,Py-CH₂),1.83(3H,s,17-CH₃),1.70(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.25(4H,m,Py-CH₂)；¹³C-NMR:182.3,160.2,158.0,156.4,153.9,151.6,149.6,138.4,131.9,127.2,123.1,116.1,115.7,110.6,104.5,104.0,94.2,78.1,61.6,46.7,46.6,28.4,28.4,26.4,25.8,25.0,25.0,18.2.

Example 21: synthesis of alpha-azadirachtin derivative HZAM-21 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, 0.8 equivalent of cyclylamine, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, slowly dropwise adding is carried out under ice bath, after the dropwise adding is finished, an deicing salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, suction filtering and concentrating to obtain a crude product of yellow oil, and then treating the crude product with petroleum ether: the volume ratio of ethyl acetate is 20: 1, eluting with mixed solvent, detecting with thin layer, collecting eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing eluates, and evaporating under reduced pressure to remove eluateThe product obtained by the preparation is dried to obtain HZAM-21 with the yield of 94 percent.

HZAM-21:ESI-MS m/z(％):530([M+Na]⁺,C₂₉H₃₃NO₇)；¹H-NMR(CDCl₃500MHz)13.56(1H,s,5-OH),7.19(1H,s,H-10),6.73(1H,d,J＝10.1Hz,H-4),6.25(1H,s,H-12),5.58(1H,d,J＝10.1Hz,H-3),5.27(1H,m,N-H),5.24(1H,s,H-14),4.15(1H,s,H-13),4.14(1H,s,H-13),3.78(3H,s,8-OCH₃),3.32(2H,m,Bu-CH₂),1.85(3H,s,17-CH₃),1.70(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.60(2H,m,Ar-CH₂),1.41(2H,m,Bu-CH₂),1.26(2H,m,Bu-CH₂),0.99(3H,m,Bu-CH₃)；¹³C-NMR:182.3,160.3,158.0,156.4,153.9,146.8,138.5,132.0,127.2,123.0,116.2,115.6,110.4,104.5,104.0,94.2,78.1,61.6,41.2,31.8,28.4,28.4,26.4,25.8,19.9,18.2,13.7.

Example 22: synthesis of alpha-azadirachtin derivative HZAM-22 (FIG. 1 shows the scheme)

The synthesis of compound HZA was as described above.

BTC (58mg,0.2mmol) is weighed into a 25mL two-neck flask, 5mL of anhydrous dichloromethane is added, magnetic stirring is carried out, equivalent n-butylamine, triethylamine (0.25mmol) and 5mL of dichloromethane are added into a 25mL constant pressure dropping funnel, dropwise addition is slowly carried out in an ice bath, after the dropwise addition is finished, an ice salt bath is removed, and stirring is carried out for 4 hours at room temperature.

HZA (40mg,0.1mmol) was added to a 10mL single-neck flask, a trace amount of DMAP, 5mL of dichloromethane were added, and the mixture was magnetically stirred at room temperature until complete dissolution. And slowly dripping the solution into the reaction solution in the previous step under an ice salt bath, withdrawing the ice salt bath after dripping is finished, and stirring at room temperature. After the reaction is finished, adding distilled water with the volume of 3-6 times of that of the reaction solution into the reaction solution, extracting for 3-5 times by using ethyl acetate, combining organic layers, washing by using a saturated NaCl solution, and using anhydrous Na₂SO₄Drying, vacuum filtering, concentrating to obtain yellow oily crude product, and treating with petroleumEther: the volume ratio of ethyl acetate is 20: 1, eluting with a mixed solvent, detecting with a thin layer, collecting the eluate with strong ultraviolet at 254nm and polarity less than HZA, mixing the eluates, evaporating under reduced pressure to remove the eluting solvent to obtain a product, and drying to obtain HZAM-22 with yield of 94%.

HZAM-22:ESI-MS m/z(％):556([M+Na]⁺,C₃₁H₃₅NO₇)；¹H-NMR(CDCl₃)13.57(1H,s,5-OH),7.20(1H,s,H-10),6.74(1H,d,J＝10.1Hz,H-4),6.26(1H,s,H-12),5.59(1H,d,J＝10.1Hz,H-3),5.22(1H,s,H-14),5.14(1H,m,N-H),4.15(1H,s,H-13),4.14(1H,s,H-13),3.78(3H,s,8-OCH₃),3.54(1H,m,He-CH),2.01(2H,m,He-CH₂),1.93(2H,m,He-CH₂),1.85(3H,s,17-CH₃),1.70(3H,s,16-CH₃),1.48(6H,s,18,19-CH₃),1.42(2H,m,He-CH₂),1.39(2H,m,He-CH₂)；¹³C-NMR:182.3,160.3,158.0,156.4,153.9,152.1,146.8,138.5,132.0,127.2,123.0,116.2,115.6,110.4,104.5,104.0,94.2,78.1,61.6,50.5,33.2,,28.4,28.4,26.4,25.9,24.9,24.7,18.2.

Example 23: therapeutic effect of HZAM-1 on vascular dementia

Preparation of vascular dementia rat model: male SD rats (10-12 weeks, 220- & 250g, 10 per group) were randomly grouped and acclimatized 7 days prior to the start of the experiment to create the MCAO model using the modified wire-tying method of Zea Longa et al and Nagasawa et al. Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (1g/kg) and fixed supine. Separating the right Common Carotid Artery (CCA), the Internal Carotid Artery (ICA) and the External Carotid Artery (ECA), hanging the threads for standby, ligating the ECA and the CCA, clamping the ICA distal end by an artery clamp, quickly making an incision at the bifurcation of the ECA and the ICA, inserting one end of the incision and heating the other end to form a smooth spherical nylon thread plug. After inserting the thread into ICA, slightly ligating the nylon thread and ICA section at the entrance, then loosening the artery clamp for clamping ICA, continuously inserting the nylon thread until the nylon thread has slight resistance and then slightly withdrawing until the thread insertion depth is about (18.5 +/-0.5) mm, and realizing cerebral ischemia caused by middle cerebral artery occlusion. The entrance was ligated again, leaving about 1cm of nylon thread outside, and the skin was sutured. And after 45min, gently lifting the remained thread ends until resistance exists, realizing the refilling of the middle cerebral artery, and finishing the molding. Only ECA and ICA were ligated in the sham group. Animal model success standard (animals scored 2-3 according to Zea Longa 5 grade scoring method) animal model success is judged if contralateral forelimb fatigue or walking and circling or walking and side-to-side falling signs appear after 45min of ischemia, and animals without signs or still awake after operation are regarded as unsuccessful model and are rejected. CON group was sham operated, i.e. placebo; VD is the model set. HZAM-1 with the purity of 99 percent is dissolved in isopropyl myristate for injection, and animals are administrated by intraperitoneal injection, and the dosage is as follows: 5mg/kg, 10mg/kg, 20mg/kg, and the CON group and the model group were injected intraperitoneally with a normal saline containing an equal amount of isopropyl myristate for injection. The administration was 19:00 days, and continued for 20 days.

Effect of HZAM-1 administration on the Morris water maze test in rats with vascular dementia: the water maze consists of a pool with the diameter of 150cm, the height of 50cm and the water temperature of 20 ℃, a camera and an analysis system. The water depth of the pool is 30cm, and the circular water surface is divided into 4 quadrants by two imaginary vertical lines passing through the circle center. An escape platform with the diameter of 10cm is placed at the position 1.5cm away from the water surface in the middle of the water area of one quadrant. The platform is made of transparent materials so as to prevent the experimental rat from directly observing the platform. The rats are randomly placed into water from four quadrants of 1, 2,3 and 4 of the pool, the rats can try to escape from the water environment, when the rats inhabit on the platform after the platform is found, the latency time of the rat finding platform is recorded by an automatic tracking system, if the latency time exceeds 90s, the latency time is recorded as 90s, and the average latency period of each day is calculated. The experimental result shows that the escape latency of the rats in the model group is obviously longer than that in the sham operation group (P is less than 0.05) in the test days 1 to 4, and the escape latency is obviously reduced in the administration group of 5mg/kg on the

days

1, 2,3 and 4 (P is less than 0.05); the 10mg/kg administration group significantly reduces the escape latency (P < 0.05) on

days

1, 2 and 4; the group of 20mg/kg significantly reduces the escape latency (P < 0.05) on

days

1, 2,3 and 4, and the swimming speed of the mice does not change obviously within 4 days. As shown in figure 1. The result shows that HZAM-1 can obviously improve the problem of low learning and memory caused by vascular dementia and can obviously improve the learning and memory ability of mammals.

Effect of HZAM-1 administration on the open field space exploration behavioral test in vascular dementia rats: the open field experimental device consists of an open field experimental box and an ANY-maze video tracking system. Thoroughly clear up spacious field experimental box before the experiment, confirm that it is clean tasteless. The experimenter records the serial number of the rat in the operation software, holds the tail 1/3 of the rat and puts the rat into one corner of the experimental box facing the box wall, the experimenter quickly starts the recording system and evacuates to a place where the experimental rat cannot see, and the recording time of the experiment is 3 minutes. After the experiment, the rats were returned to the cages, the experimental box was cleaned and wiped with 75% alcohol, and the next experiment was performed after air-drying. The rats are shuttled in the middle of the open field, the front arms are lifted, the exploration behaviors of the rats in the strange environment can be regarded as that each rat is placed in the open field for 120s, and the times of crossing the grids, the times of lifting the front arms and the residence time of the central grid are recorded. The experimental result shows that the quarts times of rats in the model group are obviously less than those in the false operation group (P < 0.05) in the test period of 1 to 3 days, and the quart times (P < 0.05) are obviously increased in the administration group of 5mg/kg on the

days

1, 2 and 3; the 10mg/kg administration group has a remarkable increase of quart times (P < 0.05) on

days

1, 2 and 3; the 20mg/kg administration group showed a significant increase in quart number (P < 0.05) on

days

1, 2 and 3. The forearm lifting frequency of the rats in the model group is obviously less than that in the sham operation group (P is less than 0.05) in the test 1 to 3 days, and the forearm lifting frequency of the rats in the administration group of 5mg/kg is obviously increased in the 1 st, the 2 th and the 3 rd days (P is less than 0.05); the administration group 10mg/kg significantly increases the forearm lifting times on

days

1, 2 and 3 (P is less than 0.05); the 20mg/kg administration group significantly increased forearm lifting times on

days

1, 2 and 3 (P < 0.05). The residence time of the central grid of the rats in the model group is obviously less than that in the sham operation group (P < 0.05) in the test 1 to 3 days, and the residence time of the central grid is obviously increased in the administration group of 5mg/kg on the 1 st, the 2 rd and the 3 rd days (P < 0.05); the 10mg/kg administration group significantly increases the median residence time (P < 0.05) on

days

1, 2 and 3; the 20mg/kg group significantly increased the median residence time (P < 0.05) on

days

1, 2, and 3. As shown in fig. 2,3 and 4. The result shows that HZAM-1 can obviously improve the adaptability of the vascular dementia animals to the new and different environments and help to recover the exploration behaviors of learning.

Results of observation of pathological changes in CA1 region of hippocampal dentate gyrus of rats with vascular dementia by HZAM-1 administration: after anesthesia of rat chloral hydrate, brain tissue was isolated after perfusion with 4% paraformaldehyde (0.1M phosphate buffer, PH 7.4), immersed in 4% paraformaldehyde, stored at 4 ℃ in the dark for 24 hours, further immersed in 30% sucrose solution for 3 days, and sectioned with a microtome. Freezing, slicing, air drying, adding anhydrous alcohol for 5-10min, adding staining solution dropwise into 75% alcohol for 3min, adding 95% alcohol twice, adding xylene twice for 5min, sealing with neutral gum, air drying, staining cells in blue-purple color, and taking picture with microscope. The experimental results showed that the cells in the sham-operated group were closely arranged and well shaped, as shown in FIG. 5. The VD model group showed obvious cell loss and scattered cell arrangement, as shown in FIG. 6. The administration group was improved more than the model group, and the cells were arranged more regularly, as shown in FIG. 7. The hippocampal dentate gyrus CA1 region plays a role in memory and spatial localization, where vascular dementia causes apoptosis and loss of cells. The results show that HZAM-1 can improve the state of hypomnesis and directional perception loss of vascular dementia animals by reversing damaged cellular substructure of the CA1 region of the hippocampal dentate gyrus.

Example 24: therapeutic effect of HZAM-1 on cerebral infarction

90 rats were randomly divided into 6 groups, namely a sham operation group, a model group, an edaravone group (3mg/kg, 0.02mmol), a butylphthalide group (20mg/kg, 0.1mmol), and a HZAM-1 group (60mg/kg,0.1mmol), and 15 rats were each. A rat focal cerebral ischemia reperfusion injury model is prepared by adopting a Longa line embolism method, so that cerebral ischemia caused by cerebral middle artery occlusion is realized. And after 2h, realizing middle cerebral artery refilling, and finishing molding. Sham groups isolated only CCA, ICA and ECA without any treatment. After the rat revives, screening out a model of 2 points or 3 points according to the Zea longa 5 division standard and entering the subsequent experiment. After 2h of ischemia, the medicine is respectively administered once in the 0h, 6h and 12h after blood reperfusion, and the medicine with the corresponding dose in the corresponding group is injected into the abdominal cavity. The pseudosurgery group and the model group are injected with isopropyl myristate solvent in the abdominal cavity, and the edaravone group is injected with edaravone, butylphthalide and HZAM-1 with corresponding doses in the abdominal cavity. The administration volume of each group was 1ml/100 g. The rats were anesthetized again after 22h of reperfusion, and brain tissue was taken after the left ventricle was perfused with PBS to wash out blood for rat brain tissue moisture content and cerebral infarction area measurements, with the results shown in table 1 and table 2. Detecting the content of tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) in serum by ELISA method, and using biochemical reagentBox index detection of calcium assay content (Ca) in brains of rats of each group²⁺) And Nitric Oxide (NO) content determination. The results are shown in Table 3. In the aspect of cerebral infarction area, compared with a sham operation group, the cerebral infarction area of a model group rat is obviously increased (P is less than 0.01); compared with the model group, the cerebral infarction areas of rats in the butylphthalide group and the HZAM-1 group are obviously reduced (P is less than 0.01), and the edaravone has no obvious effect. Compared with edaravone, HZAM-1 has obviously better effect than edaravone, and has no obvious difference with butylphthalide. In the aspect of the water content of the brain tissue of the rat, compared with a sham operation group, the water content of the brain tissue of the rat in a model group is obviously increased (P is less than 0.01); compared with the model group, the water content of the brain tissue of rats in the edaravone group and the HZAM-1 group is remarkably reduced (P is less than 0.01). But butylphthalide has no obvious effect. Compared with butylphthalide, HZAM-1 has obviously better effect than butylphthalide, and has no significant difference with edaravone. Experimental results show that HZAM-1 has obvious effects on improving the cerebral infarction area and the water content of the brain tissue of a rat, and butylphthalide and edaravone only have effects on improving the cerebral infarction area or the water content of the brain tissue of the rat. This indicates that HZAM-1 ischemic brain injury has multiple effects, indicating that it has better clinical therapeutic effects than butylphthalide and edaravone. The kit is adopted to detect related indexes and discover HZAM-1 to TNF-alpha, IL-1 beta and Ca²⁺Both MDA and butylphthalide had good efficacy, while both butylphthalide and edaravone had only partial efficacy on the index. The result is also identical with the whole animal experiment result, and the reason that the HZAM-1 has obvious effect on improving the cerebral infarction area and the water content of the brain tissue of the rat is well explained. Meanwhile, the ischemic brain injury is a disease caused by multiple factors, and a good treatment effect is difficult to achieve only by aiming at a certain treatment target. In summary, HZAM-1 has good effect on multiple target spots causing ischemic brain injury, and has good clinical application prospect.

TABLE 1 Effect on cerebral infarct size in rats

Group of	Percentage of infarct area (%)
		Artificial operation group	0.00±0.00
Model set	15.91±9.06^a
		Edaravone group	6.44±5.15
Butylphthalide group	1.64±1.03^b
		HZAM-1 group	1.78±0.91^bc

^aCompared with a sham operation group, P is less than 0.01;^bp is less than 0.01 compared with the model group;^cp is more than 0.1 compared with butyl phthalein group;

TABLE 2 Effect on Water content of rat brain tissue

Group of	Water content (%)
		Artificial operation group	77.89±0.48
Model set	83.17±2.40^a
		Edaravone group	80.33±1.29^b
Butylphthalide group	81.20±3.26
		HZAM-1 group	77.41±0.64^bc

^aCompared with a sham operation group, P is less than 0.01;^bp is less than 0.01 compared with the model group;^ccompared with edaravone, P is more than 0.1

TABLE 3 TNF-alpha, IL-1 beta, Ca in rat tissues²⁺MDA content

^aCompared with a sham operation group, P is less than 0.01;^bp is less than 0.01 compared with the model group;^cp is greater than 0.1 compared to the model group;^dp is less than 0.0 compared with the model group.

Claims

1. The application of the alpha-toosendanin derivative shown as the formula (I) in preparing the medicine for treating the brain injury disease caused by ischemia is characterized in that: the brain injury diseases are cerebral infarction and vascular dementia;

in the formula (I), R₁Is C₁-C₆A linear or branched alkyl group; r₂Is a phenyl group or a substituted phenyl group,the substituent of the substituted phenyl is methyl, ethyl, chlorine, bromine, fluorine, methoxy, amino or nitro.

2. The use of claim 1, wherein: the vascular dementia is angiopathy, learning and memory disorder caused by cerebral ischemia or cell loss in dentate gyrus region of hippocampus.