CN110433168B

CN110433168B - Application of cornuside in preparation of medicine for treating Alzheimer's disease

Info

Publication number: CN110433168B
Application number: CN201910834734.2A
Authority: CN
Inventors: 张维库; 续洁琨; 贺晓丽; 赫军; 连雯雯; 王泽星
Original assignee: China Japan Friendship Hospital
Current assignee: China Japan Friendship Hospital
Priority date: 2019-09-05
Filing date: 2019-09-05
Publication date: 2021-07-09
Anticipated expiration: 2039-09-05
Also published as: CN110433168A

Abstract

The invention relates to a new application of secoiridoid derivatives in the field of pharmacy, in particular to an application of secoiridoid derivatives in preparing medicines for preventing and treating neurodegenerative diseases. The product can obviously increase the activity of nerve cells and play an obvious role in neuroprotection in the treatment of neurodegenerative diseases, has stronger protective effect on SH-SY5Y cell models damaged by hydrogen peroxide than loganin and morroniside under the same molar concentration, can obviously improve the learning and memory disorder of aging individuals, and obviously improve the total antioxidant capacity in brain tissues.

Description

Application of cornuside in preparation of medicine for treating Alzheimer's disease

Technical Field

The invention relates to application of a secoiridoid derivative, in particular to application of the secoiridoid derivative in preparing a medicament for preventing and treating neurodegenerative diseases.

Background

Alzheimer's Disease (AD) is a serious chronic neurodegenerative disease, occurring mainly in the elderly, and the main pathological changes in the brain of patients are deposition of amyloid plaques (SP) in the forebrain basement, hippocampus and cerebral cortex, neurofibrillary tangles (NFT), loss of neuronal and neuronal synapses, and decreased cholinergic function.

The medicine for treating neurodegenerative diseases mainly comprises antioxidant, antiinflammatory agent, and anti-apoptosis agent. Among them, most of the therapeutic agents for alzheimer's disease are cholinesterase inhibitors, such as tacrine, donepezil, and rivastigmine.

The traditional Chinese medicines have long-term clinical application history in the aspect of preventing, relieving or treating neurodegenerative diseases such as Alzheimer's disease and the like, have definite curative effects and small adverse reactions, and increasingly become hot spots of research of people. Researches find that the traditional Chinese medicines such as ginseng, ginkgo leaf, morinda officinalis, shinyleaf yellowhorn and the like and extracts thereof have the obvious effect of relieving or treating neurodegenerative diseases such as Alzheimer's disease and the like. Therefore, it is of great significance to search and develop new drugs for preventing, relieving or treating neurodegenerative diseases such as Alzheimer's disease from traditional Chinese medicines.

Secoiridoid compounds such as Cornuside (Cornuside) are a class of monoterpene compounds. Research shows that the cornuside has the effects of reducing blood sugar (J Agr Food chem.2011,59 (14)), 7743-.

At present, regarding ring-splitting iridoid compounds, Chinese patent applications CN105017353A and CN105017353A disclose preparation methods, Chinese patent application CN101704857A discloses separation and purification processes, and Chinese patent applications CN106680399A, CN104991002A, CN105486775A and CN104147079A disclose content determination methods, and any report on the aspect of resisting neurodegenerative diseases such as Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis, multiple sclerosis and the like is not found in any report.

Disclosure of Invention

The invention aims to provide a new application of secoiridoid derivatives, namely a new application in pharmacy.

In fact, the present invention relates to the use of secoiridoid derivatives in the prevention or treatment of neurodegenerative diseases.

The neurodegenerative disease includes Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and multiple sclerosis.

The inventor takes cornuside, morroniside and loganin as examples, and researches a comparative experiment of the influence of secoiridoid derivatives on SH-SY5Y nerve cells damaged by hydrogen peroxide. On the basis, a comparative experiment of dogwood new glycoside, morroniside, loganin and dogwood total glycoside on learning and memory ability of D-galactose-induced aging mice is further considered. The comparison experiment results show that the protective effect of the cornus officinalis new glycosides on the neurodegenerative diseases is greatly superior to that of morroniside, loganin or cornus officinalis total glycosides, and the cornus officinalis new glycosides can be directly used as raw material medicines and further processed into medicines for preventing and treating the neurodegenerative diseases such as Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis, multiple sclerosis and the like.

In the present invention, said secoiridoid derivative preferably has the following chemical formula or a pharmaceutically acceptable salt of said compound I:

wherein R is₁And R₂Each independently selected from H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aryl.

In some embodiments, R1 is selected from H,

Wherein, X is halogen (such as F, Cl, Br and I), and n is an integer between 1 and 10.

In some embodiments, R₂Selected from H, methyl or CH₃CH₂CH＝C(CH₃)CH₂-。

In the present invention,

refers to the end of the substituent attached to the parent nucleus structure.

In some embodiments, the secoiridoid derivatives referred to in the present invention include the following compounds (but are not limited to the following compounds):

the number C1 is cornuside, and the number C2-C6 is cornuside derivative.

Through a series of cell experiments, animal behavior experiments, enzyme activity determination and the like, the compound C1-C6 is found to show remarkable anti-oxidative stress induced nerve cell injury activity, wherein the activity of cornuside is the best.

The invention also provides a pharmaceutical composition containing the secoiridoid derivative.

In some embodiments, the secoiridoid derivative is of the general chemical formula or a pharmaceutically acceptable salt of said compound I:

wherein R1 and R2 are each independently selected from H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aryl;

preferably, R₁Selected from H,

Wherein X is halogen, and n is an integer between 1 and 10;

preferably, R₂Selected from H, methyl or CH₃CH₃CH＝C(CH₃)CH₂-。

Preferably, n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

The pharmaceutically acceptable salts of the present invention are mainly inorganic base salts or organic base salts of compound I, such as inorganic ammonium salts or organic amine salts.

In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

The type and amount of the pharmaceutically acceptable carrier are determined mainly by the dosage form of the pharmaceutical composition, and can be determined by considering factors such as cost, administration route, bioavailability and the like.

Preferably, the dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquid, granules, dripping pills or pellets.

In some embodiments, the carrier comprises at least one of hydroxypropyl methylcellulose, a water-soluble filler, and a lubricant;

preferably, the water soluble filler comprises one or more of lactose, sucrose and mannitol;

the lubricant is one or more of magnesium stearate, micropowder, silica gel and talcum powder.

The content of hydroxypropyl methylcellulose in the pharmaceutical composition is 65.5-82.6 wt%.

Preferably, the weight ratio of the secoiridoid derivative to hydroxypropyl methylcellulose is 1:2 to 1:5, including but not limited to 1:2, 1:3, 1:4, 1:5, and the like.

Preferably, the content of the secoiridoid derivative in the pharmaceutical composition is 16.5-32.8 wt%; preferably, the content of the lubricant in the pharmaceutical composition is 0.6-1.3 wt%.

Particularly, when the pharmaceutical composition is prepared into tablets, better drug effect can be achieved by adopting the carrier and the mixture ratio.

The invention relates to application of homosecoiridoid derivatives, in particular to a pharmaceutical composition which is mainly used for preventing or treating neurodegenerative diseases.

In order to better understand the essence of the present invention, the pharmacological test and results of cornuside will be used to illustrate its new application in preparing medicine for preventing and treating neurodegenerative diseases.

Test example 1:

protective effect of Cornuside on oxidative stress-induced SH-SY5Y nerve cell injury

Human neuroblastoma (SH-SY5Y)Cells were cultured in complete DMEM/F12 medium (containing 10% fetal bovine serum, 100U/mL penicillin, 100. mu.g/mL streptomycin, 1% glutamine) at 37 ℃ in 5% CO₂Culturing under saturated humidity condition. Changing the solution every 2-3 days. Taking cells in logarithmic growth phase, adding 0.25% trypsin for digestion, diluting the cell concentration to 105 cells/mL by using a complete culture medium, inoculating the cells into a 96-well cell culture plate, culturing for 24 hours, pretreating the cells for 2 hours by using cornuside (0.3, 1 and 3 mu M) and a positive drug Vitamin E (VE)10 mu M, treating SH-SY5Y cells for 12 hours by using hydrogen peroxide (280 mu M), detecting the cell viability by using a CCK8 method after the cell treatment is finished, and evaluating the drug effect by using indexes such as LDH and NO.

As shown in FIG. 1, no toxic effect on SH-SY5Y cells was observed at (0.03-30) μ M of cornuside.

As shown in fig. 2, in the experiment for measuring cell viability by the CCK8 method, the cell viability of the model group is significantly reduced compared with that of the normal group (P < 0.01); compared with a model group, the survival rate of cells of the cornus officinalis neoside administration group is obviously improved, the obvious neuroprotective effect is shown, the maximum injury resistance effect (P is less than 0.01) is achieved at 1 mu M, and the protection rate is close to 80%.

The experimental results are shown in fig. 3, in the LDH determination experiment, the LDH leakage of the model group was significantly increased (P <0.01) compared with the normal group; compared with the model group, the LDH leakage of the cornus officinalis neoside administration group is remarkably reduced and reaches the lowest level (P <0.01) at 1 mu m.

The experimental result is shown in fig. 4, in the NO content determination experiment, the NO content of the model group is obviously increased (P <0.01) compared with the normal group; compared with the model group, the NO content of the dogwood neoside administration group is remarkably reduced (P < 0.01).

The above experimental results show that: the cornuside can increase the activity of nerve cells and play an obvious role in neuroprotection.

Test example 2:

comparative experiment of protective effect of cornuside, morroniside and loganin on SH-SY5Y cells damaged by hydrogen peroxide

The experimental results are shown in fig. 5, the cornuside and morroniside both can generate protective effect on SH-SY5Y cells damaged by hydrogen peroxide, wherein the protective effect of the cornuside on SH-SY5Y cell models damaged by hydrogen peroxide at 0.3 mu M and 1 mu M is stronger than that of the morroniside under the same molar concentration.

The experimental results are shown in FIG. 6, the protective effect of cornuside and loganin on SH-SY5Y cells damaged by hydrogen peroxide can be achieved, wherein the protective effect of cornuside on SH-SY5Y cell models damaged by hydrogen peroxide at 0.3 mu M and 1 mu M is stronger than that of loganin at the same molar concentration.

Test example 3:

a comparison experiment of the protective effect of cornuside, morroniside, loganin and total cornuside on the learning and memory ability of a D-galactose-induced aging mouse is carried out.

D-galactose-induced aging mouse model preparation:

neck and back subcutaneous injection of D-galactose 100mg kg^-1And establishing an aging model in 8 weeks.

Grouping and administration:

healthy 2-month-old KM mice were randomly divided into 6 groups of 12 mice (male and female halves) as normal groups, i.e., D-galactose-induced aging group, cornus officinalis neoside group (purity of 95% or more, self-made), morroniside group, loganin group, and cornus officinalis total side group.

Normal group: the KM mice are subjected to intragastric administration and double-distilled water with the same amount every day, and physiological saline with the same amount is injected subcutaneously at the neck and the back;

model group: the KM mice were gavaged with equivalent double distilled water every day, and injected with 100 mg/kg of D-galactose subcutaneously at the back and neck^-1。

Cornus officinalis neo-glycosides group: daily gavage administration of cornuside to KM mice at a dose of 30mg/kg ^-1100 mg/kg of D-galactose for subcutaneous injection at the back and neck^-1。

Morroniside group: the KM mice are administrated with morroniside by intragastric administration every day, and the dose is 22mg kg^-1(different molecular weights, based on the molecular weight of the same molar concentration), the neck and back were injected subcutaneously with 100mg kg of D-galactose^-1。

Loganin group: daily gavage administration of loganin to KM mice at a dose of 21 mg/kg^-1(different molecular weights, depending on the molecule)Amount converted to equivalent molar concentration), 100 mg/kg of D-galactose was injected subcutaneously into the back and neck^-1。

The dogwood total glycosides group (the source is that dried and mature fruits of dogwood are heated and refluxed to extract, and then are absorbed by D101 macroporous absorption resin after being decompressed and concentrated, and then are eluted by 15 percent ethanol and 35 percent ethanol in sequence, and the 35 percent ethanol eluate is merged, decompressed, concentrated and dried to obtain the dogwood total glycosides, and the chemical analysis shows that the dogwood total glycosides group mainly comprises loganin and morroniside): feeding Corni fructus total glycosides into KM mouse by intragastric administration at a dose of 50mg kg ^-1100 mg/kg of D-galactose for subcutaneous injection at the back and neck^-1。

Results and analysis:

the degree of memory impairment of the D-galactose model and the effect of the drugs on impairment improvement were assessed by behavioral indicators (water maze, new object recognition) 6 weeks after administration. The longer the latency of the model group, the fewer the platform-crossing times and the poorer the object recognition capability, which indicates that the learning and memory function of the animal is more seriously damaged. The shorter the incubation period of the administration group, the more the number of times of platform crossing, the stronger the object recognition ability, and the stronger the improvement effect of the drug.

Table 1 shows the results of comparing the effects of cornin, morroniside, loganin and dogwood total glycosides on the navigation ability of D-galactose-induced aging mice.

TABLE 1

Table 2 shows the results of comparing the effect of cornin with morroniside, loganin and total cornin on the spatial exploration ability (latency) of D-galactose-induced aging mice.

TABLE 2

Table 3 shows the results of comparing the effect of cornin with morroniside, loganin and total cornin on the spatial exploration ability (number of crossing platforms) of D-galactose-induced aging mice.

TABLE 3

Table 4 shows the results of comparison of the effects of cornin, morroniside, loganin and total cornin on the discrimination index in the D-galactose-induced aging mouse new body recognition experiment.

TABLE 4

The experimental results show that in the positioning navigation experiment, the incubation period of the mice of each experimental group shows a shortening trend along with the increase of the training times and the experimental days, and the results are shown in table 1. In the space exploration experiment, compared with the normal group of mice, the space exploration capacity of the model group of mice is reduced, which is shown in that the time for the first time to pass through the platform is prolonged (P <0.05), and the results are shown in Table 2. The number of platform crossings was significantly reduced (P <0.05) and the results are shown in table 3. Compared with a model group, the dogwood neoside can obviously improve the learning and memory ability of mice, and is represented by that the first-time platform crossing time is reduced (P <0.05), the platform crossing times are increased, and the dogwood neoside has significant difference (P < 0.01). In the new object identification experiment, compared with the normal group of mice, the discrimination index of the model group of mice is reduced and has significant difference (P <0.01), compared with the model group, the discrimination index of the mice can be obviously improved by the cornuside, and has significant difference (P <0.01), and the result is shown in table 4.

The above experimental results show that, in the experiment of mouse aging caused by D-galactose, 30mg/kg of cornuside is administered by gastric gavage^-1The mouse can obviously improve the learning memory disorder of mice, including space memory disorder and scene memory disorder. Compared with administration of morroniside, loganin and Corni fructus total glycosides by intragastric administration, cornuside 30mg/kg^-1Can more obviously increase the learning and memory ability of the mouse and show more effective pharmacological action.

Test example 4:

effect of cornuside on D-galactose-induced oxidative stress injury of brain tissue of aged mice

Immediately after the behavioral experiments (week 8), the mice were sacrificed, and hippocampal and cerebral cortical proteins were separated from precooled physiological saline and frozen in a freezer at-80 ℃. Weigh skin layer before measurement, 1: 9 adding normal saline to prepare 10% homogenate at 4 deg.C, 3500 r.min^-1Centrifuging for 10min, collecting supernatant, and freezing at-80 deg.C in refrigerator for subsequent determination of oxidative stress indexes such as acetylcholinesterase (AChE), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC).

The experimental results are shown in fig. 7, and the total antioxidant capacity of the model group is significantly reduced compared with that of the normal group (P < 0.01). Compared with the model group, the total antioxidant capacity of the brain tissue of the mice of the cornuside administration group is obviously improved.

The experimental result is shown in fig. 8, compared with the normal group, the glutathione peroxidase level of the model group is obviously reduced (P < 0.01); compared with the model group, the glutathione peroxidase level in the corresponding brain tissue of the dogwood neoside administration group is obviously increased (P < 0.01).

The experimental result is shown in fig. 9, compared with the normal group, the acetylcholinesterase level of the model group is obviously increased (P < 0.01); compared with the model group, the brain tissue corresponding to the dogwood neoside administration group has obviously reduced acetylcholinesterase level (P < 0.05).

Through a series of activity screening experiments of resisting nerve cell injury, the compound C1-C6 shows remarkable activity of resisting the nerve cell injury induced by oxidative stress, wherein the activity of cornuside is the best.

The research of the invention shows that the secoiridoid derivative can obviously improve the learning and memory ability of mice, and is represented by the fact that the time for passing through a platform for the first time is reduced and the frequency for passing through the platform is increased in a space exploration experiment; in a new object identification experiment, the discrimination index of a mouse is obviously improved; the total antioxidant capacity in the brain tissue of the mouse is obviously improved; a significant reduction in acetylcholinesterase levels; in addition, the compound can effectively inhibit the damage of the neuron cells caused by the hydrogen peroxide. In conclusion, the secoiridoid derivative can be prepared into medicines or health care products for resisting neurodegenerative diseases such as Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis, multiple sclerosis and the like.

In summary, compared with the prior art, the invention achieves the following technical effects:

(1) the invention has explored new medical application to secoiridoid derivative, has opened up a new application field;

(2) the secoiridoid derivative is safe and nontoxic, has strong pharmacological action and indicates good medicinal prospect;

(3) the product of the invention has the advantages of raw material sources of natural plants, low price, no toxic or side effect, simple preparation process, capability of being prepared into oral dosage forms, injection dosage forms, tablets and the like, and convenient use;

(4) the medicine prepared by the product can increase the activity of nerve cells and play an obvious role in protecting nerves;

(5) the protection effect of the medicine prepared by the product on the SH-SY5Y cell model damaged by hydrogen peroxide is stronger than that of loganin and morroniside under the same molar concentration;

(6) the medicine prepared by the product can obviously improve the learning and memory disorder of an aged individual, is superior to morroniside, loganin and dogwood total glycoside, and obviously improves the total antioxidant capacity in brain tissues.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the invention and, together with the description, serve to explain the invention and not to limit the invention. Wherein:

FIG. 1 shows the neurotoxic effect of cornuside on SH-SY5Y cells;

FIG. 2 effect of cornuside on the cell viability of SH-SY5Y cells damaged by hydrogen peroxide;

FIG. 3 is the effect of cornuside on LDH release from SH-SY5Y cells damaged by hydrogen peroxide;

FIG. 4 is the effect of cornuside on the NO release of SH-SY5Y cells damaged by hydrogen peroxide;

FIG. 5 is a comparative experiment of the effect of cornuside and morroniside on the survival rate of SH-SY5Y cells damaged by hydrogen peroxide;

FIG. 6 is a comparative experiment of the effect of cornuside and loganin on the survival rate of SH-SY5Y cells damaged by hydrogen peroxide;

FIG. 7 is a graph showing the effect of cornuside on the level of T-AOC in the brain of mice aged with D-galactose;

FIG. 8 is a graph showing the effect of cornuside on the levels of GSH-Px in brains of mice aged by D-galactose;

FIG. 9 the effect of cornuside on the levels of AChE in brains of mice aged by D-galactose.

Detailed Description

Several embodiments of the invention will be described below, but the content of the invention is not limited thereto at all.

Example 1:

a tablet containing cornuside as raw material medicine is provided.

Every 1000 tablets comprise the following components:

15g of cornuside, 30g of hydroxypropyl methylcellulose, 0.4g of talcum powder, 0.2g of lactose, 0.2g of magnesium stearate and a proper amount of absolute ethyl alcohol, and the cornuside, the hydroxypropyl methylcellulose, the magnesium stearate and the absolute ethyl alcohol are prepared into 1000 tablets.

When the tablet is prepared, cornuside, hydroxypropyl methylcellulose, talcum powder, lactose and magnesium stearate are uniformly mixed, absolute ethyl alcohol is added to prepare a soft material, the soft material is sieved by a 24-mesh sieve, prepared into granules, dried, added with magnesium stearate, uniformly mixed and tabletted.

Example 2 the same as example 1 except that the amount of hydroxypropylmethylcellulose was 45 g.

Example 3 the same as example 1 except that the amount of hydroxypropylmethylcellulose was 75 g.

Example 4

Capsules using cornuside as raw material medicine.

15g of cornuside, 10g of starch, 4g of dextrin, 3g of hydroxypropyl cellulose, 4g of magnesium stearate and a proper amount of absolute ethyl alcohol, and 1500 granules are prepared.

Mixing cornuside with starch, dextrin and hydroxypropyl cellulose, adding anhydrous ethanol to obtain soft material, sieving with 24 mesh sieve, granulating, drying, adding magnesium stearate, mixing, and making into capsule.

Example 5

Granules using cornuside as a raw material medicine.

15g of cornuside, 15g of cane sugar, 15g of starch, 3g of talcum powder, 7g of croscarmellose sodium, 7g of magnesium stearate and a proper amount of absolute ethyl alcohol, and the mixture is prepared into 1500 bags.

Mixing cornuside with sucrose, starch, pulvis Talci, and croscarmellose sodium, adding anhydrous ethanol to obtain soft material, sieving with 24 mesh sieve, granulating, drying, adding magnesium stearate, mixing, and packaging.

Example 6

Oral liquid taking cornuside as raw material medicine.

15g of cornuside, 3g of warfarin, 12g of cane sugar, 3g of sodium bisulfite, 3g of methyl p-hydroxybenzoate, a proper amount of sodium bicarbonate and 15000ml of water for injection, and the prepared product is 1500 pieces.

Mixing the above materials, and packaging by conventional method.

Example 7

The dripping pill takes cornuside as a raw material medicine.

15g of cornuside, 100g of polyethylene glycol 6000 and a proper amount of absolute ethyl alcohol, and are prepared into 1500 granules.

Weighing cornuside of a prescription amount, adding a proper amount of absolute ethyl alcohol, slightly heating to dissolve, adding polyethylene glycol 6000 of the prescription amount (keeping the temperature in a water bath at 60 ℃), stirring and mixing uniformly until the ethyl alcohol is volatilized completely, continuously standing in the water bath at 60 ℃ for keeping the temperature for 30min, transferring into a liquid storage barrel after bubbles are removed, controlling the dripping speed at 70-80 ℃, dripping into condensate dropwise, pouring off the condensate after complete condensation, and collecting dripping pills.

Example 8

A tablet using the compound C2 as a raw material medicine.

Every 1000 tablets comprise the following components:

compound C215 g, hydroxypropyl methylcellulose 30g, talcum powder 0.4g, lactose 0.2g, magnesium stearate 0.2g, and appropriate amount of anhydrous ethanol, and making into 1000 tablets.

When the tablets are prepared, the compound C2 is uniformly mixed with hydroxypropyl methyl cellulose, talcum powder, lactose and magnesium stearate, absolute ethyl alcohol is added to prepare a soft material, the soft material is sieved by a 24-mesh sieve, the soft material is prepared into granules, the granules are dried, the magnesium stearate is added, the mixture is uniformly mixed, and the tablets are pressed.

Comparative example 1: the same procedure as in example 1 was repeated, except that microcrystalline cellulose was used in place of hydroxypropylmethyl cellulose.

Comparative example 2: the procedure is as in example 1 except that crospovidone is used instead of hydroxypropylmethyl cellulose.

Comparative example 3: the procedure is as in example 1 except that croscarmellose sodium is used instead of hydroxypropylmethyl cellulose.

Comparative example 4: the procedure is as in example 1 except that sodium hydroxymethyl cellulose is used in place of hydroxypropyl methylcellulose.

Comparative example 5: the same procedure as in example 1 was repeated, except that povidone K30 was used instead of hydroxypropylmethylcellulose.

Comparative example 6: the same procedure as in example 1 was repeated, except that the amount of hydroxypropylmethylcellulose was changed to 25 g.

Comparative example 7: the same procedure as in example 1 was repeated, except that 80g of hydroxypropylmethylcellulose was used.

Comparative example 8: the same procedure as in example 8 was repeated, except that microcrystalline cellulose was used in place of hydroxypropylmethyl cellulose.

Dissolution rate determination experimental study of secoiridoid derivative tablets

According to the dissolution rate determination method (XC second method which is an appendix of the second part of the 2005 edition of Chinese pharmacopoeia) and using 1000 ml of water as a dissolution medium, rotating at 75 revolutions per minute, operating according to the method, taking out the solution after 15 and 45 minutes, filtering, precisely taking 2ml of subsequent filtrate, placing the subsequent filtrate in a 50ml volumetric flask, and dissolving the subsequent filtrate by using 0.1mol/L hydrochloric acid solution to obtain a test solution. 32mg was precisely measured and placed in a 50ml volumetric flask, 2ml was precisely measured and placed in a 50ml volumetric flask, and a solution of 0.1mol/L hydrochloric acid was used as a control solution. Respectively taking the test solution and the reference solution, respectively measuring absorbance at 345nm wavelength according to ultraviolet-visible spectrophotometry, and calculating the dissolution amount of each dosage, wherein the limit is 70% of the labeled amount and the dosage is in line with the regulation.

TABLE 5 dissolution rate assay results of secoiridoid derivative tablets

The experimental results are shown in table 5, the split ring iridoid derivative tablet prepared by adopting the specific disintegrant is basically completely dissolved out in 45min, the dissolution rate in 15min is more than 95%, wherein when the hydroxypropyl methyl cellulose is 30g, namely the proportion of the hydroxypropyl methyl cellulose to the cornuside or the compound C2 is 2:1, the effect of the embodiment 1 and the embodiment 8 is best; when the amount of hydroxypropylmethylcellulose is outside the range of the present invention, as shown in comparative examples 6-7, the effect is much lower than that of the examples of the present invention; when the disintegrant is other disintegrants in the field, such as the disintegrants shown in comparative examples 1-5, the compatibility of the disintegrant with the secoiridoid derivative is poor, and the dissolution rate is poor.

Research on influence of different preparations of secoiridoid derivatives on mouse new object recognition experiment

The procedure of test example 3 was followed. The KM mice were randomly divided into 18 groups, blank group, model group, preparation examples 1-8, comparative examples 1-8, and 10 mice each. The tested drugs are dissolved in distilled water according to the weight of 10ml/kg before the experiment and are administrated by intragastric administration (30 mg/kg). Mice were dosed for 30min for new object identification experiments. The experiments were performed in a quiet room at room temperature 25 ℃ between 9:00 and 17: 00.

TABLE 6 influence of different formulations of secoiridoid derivatives on mouse neologies identification experiments

The experimental results are shown in table 6, in the new body identification test, when the secoiridoid derivative tablet, capsule, granule, oral liquid and dripping pill are administrated by intragastric administration, the new body identification index (P is less than 0.05) of the mouse can be increased. In particular, when the ratio of hydroxypropyl methylcellulose to cornuside is 2:1, namely, in example 1(P <0.01), the dosage range of the secoiridoid derivative tablet prepared by using a specific disintegrant is better than that of hydroxypropyl methylcellulose (such as comparative examples 6-7), and the dosage range of the hydroxypropyl methylcellulose is better than that of capsules, granules, oral liquid and dropping pills (examples 4-7) prepared by using the secoiridoid derivative as a raw material medicament.

The above is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made to the present invention by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The application of the cornuside or the pharmaceutically acceptable salt thereof as the only active substance in the preparation of the medicine for treating the Alzheimer disease is characterized in that the cornuside has the following chemical formula:

the medicament also comprises a pharmaceutically acceptable carrier; the carrier is hydroxypropyl methylcellulose, water-soluble filler and lubricant;

the content of hydroxypropyl methylcellulose in the medicine is 65.5 wt%; the content of cornus officinalis neoside in the medicine is 32.8 wt%; the content of the lubricant in the medicine is 1.3 wt%; the content of the water-soluble filler in the medicine is 0.4 wt%.

2. The use according to claim 1, wherein the pharmaceutically acceptable salt is an inorganic or organic base salt of cornuside.

3. The use of claim 1, wherein the medicament is in the form of tablets, capsules, granules, oral liquid, dripping pills or pellets.

4. The use according to claim 1, wherein the water-soluble filler is one or more of lactose, sucrose and mannitol; the lubricant is one or more of magnesium stearate, silica gel and talcum powder.