A pharmaceutical composition for treating senile dementia

Abstract

The application belongs to the technical field of medicine, and particularly relates to a pharmaceutical composition for treating senile dementia and application thereof. The application provides a pharmaceutical composition containing (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyl formamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole and icariin. Pharmacodynamic experiments prove that the pharmaceutical composition of the application achieves a synergistic therapeutic effect in treating senile dementia.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a pharmaceutical composition for treating Alzheimer's disease, particularly a pharmaceutical composition containing (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole and icariin. Background Technology

[0002] Alzheimer's disease (AD), also known as senile dementia, is characterized by progressive memory loss and cognitive impairment. It severely reduces patients' daily activities and self-management abilities, placing a heavy burden on their families. The incidence of Alzheimer's disease remains high, and the WHO predicts that by 2050, there will be over 110 million people with the condition. The pathogenesis of Alzheimer's disease is complex and lacks a unified and clear understanding. Its contributing factors mainly include genetic factors, neurotransmitters, and immune factors. Currently, treatment for Alzheimer's disease both domestically and internationally is primarily drug-based, mainly including cholinesterase inhibitors (ChEIs) and non-competitive N-methyl-D-aspartate (NMDA) receptor inhibitors. These drugs can cause side effects on the gastrointestinal tract and central nervous system, and are prone to drug dependence, resulting in limited therapeutic efficacy. Therefore, developing a more effective drug for treating Alzheimer's disease is crucial.

[0003] Icariin is the main active ingredient of the traditional herbal medicine Epimedium, which is the dried stem and leaves of Epimedium, Epimedium sagittatum, Epimedium pubescens, or Epimedium koreanum (all belonging to the Berberidaceae family). Clinically, Epimedium is mainly used for kidney yang deficiency, impotence, frequent urination, infertility, rheumatic pain, numbness and contracture of limbs, weakness of muscles and bones, difficulty walking, cough, and shortness of breath. Before being absorbed in the body, icariin is metabolized in the intestine into icariin II and icariin aglycone, with icariin aglycone exhibiting stronger pharmacological activities than icariin II and icariin III. Both icariin and icariin have anti-neuroinflammatory, antioxidant, and anti-apoptotic effects. Icarin aglycone is mainly prepared from icariin through degradation and chemical synthesis methods.

[0004] The invention patent application with application number CN200710201297.8 discloses that icariin has a good protective effect on vascular dementia models and neurons damaged by cerebral ischemia-reperfusion injury, and can significantly improve the learning and memory performance of rat models of vascular dementia, Alzheimer's disease and other types of dementia. The pharmacological effects and mechanisms of action of icariin have been studied in depth.

[0005] In her paper "Molecular Mechanism Study of Epimedium's Anti-inflammatory Response in Alzheimer's Disease", Zhu Menglin pointed out that epimedium can inhibit LPS-induced excessive activation of microglia and improve the learning and memory abilities of mice in an AD inflammation model.

[0006] The invention patent with application number 201310362101.9 discloses that icariin can cross the blood-brain barrier, promote β-catenin translocation by inhibiting GSK-3 activity, reduce Tau protein phosphorylation, protect the vitality of neuronal cells and inhibit neurofibrillary tangles, thereby playing a role in preventing and treating Alzheimer's disease. Summary of the Invention

[0007] In order to significantly improve cognitive function in the treatment of Alzheimer's disease, the inventors, based on the latest research progress on Alzheimer's disease and the mechanism of action of icariin, creatively combined (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole (compound 1) with icariin for the treatment of Alzheimer's disease. Among them, (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole.

[0008] Pharmacodynamic experiments have demonstrated that the drug combination of the present invention synergistically improves cognitive dysfunction in AD mice, increases serum serotonin and tryptophan levels, and inhibits L-kynurenine production. Therefore, the present invention provides a novel pharmaceutical composition whose active ingredients contain icariin and compound 1.

[0009] Preferably, the weight ratio range of compound 1 to icariin in the pharmaceutical composition of the present invention can be optimized by pharmacological experiments. The weight ratio range of compound 1 to icariin is (0.5-10):1, especially when the weight ratio of compound 1 to icariin is (1.6-3.3):1, it is most effective for treating Alzheimer's disease.

[0010] Based on the properties of compound 1 and icariin, the inventors of this invention have prepared the pharmaceutical composition of this invention into a solid dosage form, wherein the tablets include ordinary tablets, coated tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, multilayer tablets, dispersible tablets, sustained-release tablets, etc. The pharmaceutical composition of this invention has the advantages of being convenient to carry, easy to administer, and readily accepted by patients.

[0011] The pharmaceutical compositions of the present invention can be prepared using conventional pharmaceutical excipients according to conventional formulation techniques. Conventional pharmaceutical excipients are selected from one or more of excipients, lubricants, binders, emulsifiers, disintegrants, stabilizers, and flavoring agents. The excipients are selected from one or more of lactose, sucrose, glucose, mannitol, sorbitol, starch, dextrin, crystalline cellulose, gum arabic, and dextran; the binders are selected from one or more of magnesium stearate, calcium stearate, talc, micronized silica gel, boric acid, and sodium lauryl sulfate; the binders are selected from one or more of hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, and polyethylene glycol; and the disintegrants are selected from low-substituted hydroxypropyl cellulose. The ingredients include one or more of carboxymethyl cellulose, sodium carboxymethyl starch, and croscarmellose; the emulsifier is selected from one or more of bentonite, magnesium hydroxide, aluminum hydroxide, and sodium dodecyl sulfate; the stabilizer is selected from one or more of methylparaben, benzyl alcohol, phenylethanol, phenol, sorbic acid, and dehydroacetic acid; and the flavoring agent is selected from one or more of sucrose, flavoring, aspartame, and cyclodextrin.

[0012] This invention investigated the effects of compound 1 and icariin on Alzheimer's disease. The results showed that the drug composition containing compound 1 and icariin in a weight ratio of (0.5-10):1 had a good effect on improving cognitive function, increasing serum 5-hydroxytryptamine levels, and inhibiting tryptophan metabolism in AD model mice, especially when the weight ratio was (1.6-3.3):1, the therapeutic effect was the best.

[0013] The pharmaceutical composition provided by this invention has the following advantages over the prior art:

[0014] 1. Synergistic technical effects have been achieved in improving cognitive function.

[0015] 2. It can significantly increase serum 5-hydroxytryptamine and tryptophan levels, inhibit tryptophan metabolism, and reduce L-kynurenine levels. Detailed Implementation

[0016] The present invention will now be further illustrated by the following embodiments, but the application scope of the present invention is not limited to the following embodiments. The following embodiments are not intended to limit the present invention in any way. For those skilled in the art, any modifications or substitutions made within the scope of the technical solution of the present invention based on the technical essence of the present invention shall still fall within the protection scope of the technical solution of the present invention.

[0017] Example 1: Ordinary tablets of the pharmaceutical composition of the present invention

[0018]

[0019] Preparation process: Weigh the prescribed amounts of icariin, compound 1, starch, and dextrin and mix them evenly. Add an appropriate amount of 50% ethanol to the mixed powder, mix evenly, form a soft mass, pass it through an 18-mesh nylon sieve to form wet granules, dry at about 60℃, the moisture content of the dry granules should be controlled below 1.5%, granulate through a 20-mesh sieve, then mix with magnesium stearate, compress into tablets, and the product is obtained.

[0020] Example 2: Capsules of the pharmaceutical composition of the present invention

[0021]

[0022] Preparation process: Icariin, compound 1, microcrystalline cellulose, and micronized silica gel are pulverized and passed through a 100-mesh sieve and mixed evenly, then directly filled into capsules.

[0023] Example 3: Dispersible tablets of the pharmaceutical composition of the present invention

[0024]

[0025] Preparation process: The prescribed amount of icariin and compound 1 are passed through a 100-mesh sieve, and calcium hydroxymethyl cellulose, cross-linked polyvinylpyrrolidone, and microcrystalline cellulose are passed through an 80-mesh sieve. After mixing, an appropriate amount of 10% starch paste is added for granulation, and magnesium stearate is added for tableting.

[0026] Example 4: Granules of the pharmaceutical composition of the present invention

[0027]

[0028]

[0029] Preparation process: Weigh the prescribed amounts of icariin, compound 1, starch, dextrin, and sucrose powder and mix them evenly. Add an appropriate amount of 80% ethanol to the mixed powder, mix evenly, form a soft mass, pass it through an 18-mesh nylon sieve to form wet granules, dry at about 60℃, granulate through a 20-mesh sieve, and package to obtain the final product.

[0030] Example 5: Sustained-release tablets of the pharmaceutical composition of the present invention

[0031]

[0032] Preparation process: Mix the prescribed amount of icariin, compound 1, hydroxypropyl methylcellulose and lactose evenly, add 5% polyvinylpyrrolidone aqueous solution to granulate, dry at 40-80℃, granulate, add the prescribed amount of micronized silica gel to the dry granules, mix well, and then press into tablets of irregular shape.

[0033] Pharmacological Example 1: Effects of the pharmaceutical composition of the present invention on Alzheimer's disease model mice

[0034] 1. Laboratory animals

[0035] One hundred SPF-grade Kunming mice, 50 males and 50 females, weighing 20-22 g, were provided by Jinan Pengyue Laboratory Animal Breeding Co., Ltd., with the animal certificate number: SCXK(Lu)20190003. The mice were housed in a SPF-grade animal room with strictly controlled environmental conditions, at a temperature of 20-26 °C, a humidity of 40-70%, a light-dark cycle of 12:12 h, and fed with growth and reproduction feed, with free access to food and water.

[0036] 2. Main instruments and reagents:

[0037] Compound 1 was provided by Shandong Xinsdailai Pharmaceutical Co., Ltd.;

[0038] Icariin was provided by Shandong Xinsdailai Pharmaceutical Co., Ltd.;

[0039] Eight-arm maze;

[0040] High performance liquid chromatography;

[0041] 3. Experimental methods and experimental groups:

[0042] First, all mice were trained in the eight-arm maze, a commonly used model for evaluating animal learning and memory abilities. After one week of acclimatization feeding, the animals were weighed and fasted for 24 hours. Thereafter, after each day's training, they were given a restricted diet to maintain their body weight at 80%–85% of that of a normally feeding rat. On the second day, food pellets (4–5 pellets per animal, approximately 3–4 mm in diameter) were scattered in each arm and the central area of ​​the maze. Then, four animals were simultaneously placed in the center of the maze (with the doors to each arm open). They were allowed free access to eat and explore for 10 minutes. On the third day, the training was repeated. This process allowed the animals to become familiar with the maze environment without significant stress. From the fourth day onwards, animals were trained individually: one food pellet was placed near the outer end of the food container in each arm, allowing the animal to eat freely. The pellet was removed after the animal had finished eating or after 10 minutes. On the fifth day, food was placed in the food container, and the training was repeated twice a day. On the sixth day, four arms were randomly selected, and one food pellet was placed in each arm. The arm doors were closed, and the animal was placed in the center of the maze. After 30 seconds, the arm doors were opened, allowing the animal to move freely in the maze and ingest the food pellets until all four arms were eaten. If the food pellets were not finished after 10 minutes, the experiment was terminated. Training was conducted twice daily, with an interval of at least one hour between sessions. Modeling began when there were zero working memory errors and no more than one reference memory error in five consecutive training sessions. Mice that met the learning criteria were selected, and 10 were randomly chosen as the normal group, 10 as the model group, and the rest as the experimental group. Except for the normal group, which received an equal volume of physiological saline intraperitoneally and an equal volume of double-distilled water by gavage, all other groups received an intraperitoneal injection of D-galactose (500 mg / kg / day) (prepared with physiological saline) and gavage of AlCl3 (20 mg / kg / day) (prepared with double-distilled water). Each group was administered the drug via gavage once daily according to the experimental grouping and dosage. The normal group and model group were administered the same volume of purified water via gavage. Administration continued for 8 weeks. The experimental mice were divided into low-dose and high-dose groups of icariin, and low-dose and high-dose groups as shown in Formula I, as well as groups A, B, C, and D, for a total of 10 groups, with 10 mice in each group. The grouping and dosage for each group are as follows:

[0043] Low-dose icariin group: Icariin 3.2 mg / (kg·d) was administered by gavage.

[0044] High-dose icariin group: Icariin 21.2 mg / (kg·d) was administered by gavage.

[0045] Low-dose group of compound 1: 10.6 mg / (kg·d) of compound I was administered by gavage.

[0046] High-dose group of compound 1: 32 mg / (kg·d) of the compound represented by formula I was administered by gavage.

[0047] Composition A: Compound 1 was administered by gavage at a dose of 5.3 mg / (kg·d) + icariin 1.6 mg / (kg·d);

[0048] Composition B: Compound 1 was administered by gavage at a dose of 5.3 mg / (kg·d) + icariin 10.6 mg / (kg·d);

[0049] Composition C: Compound 1 was administered by gavage at a dose of 16 mg / (kg·d) + 1.6 mg / (kg·d) of icariin;

[0050] Composition D: Compound 1 was administered by gavage at a dose of 16 mg / (kg·d) + 10 mg / (kg·d) of icariin;

[0051] Statistical analysis was performed using SPSS 19.0 software. Quantitative data were expressed as expressed in numerical form. t-tests and ANOVA were used, and P < 0.05 was considered statistically significant.

[0052] 3.1 Behavioral Experiments:

[0053] Each group of mice was placed in the center of an eight-arm maze for the experiment. Working memory error, reference memory error, total number of arm insertions, and test time were recorded.

[0054] The working memory error frequency, reference memory error frequency, and average exploration time were calculated based on the recorded working memory errors, reference memory errors, total number of arm insertions, and testing time.

[0055] Working memory error frequency = Number of working memory errors / Total number of arm insertions;

[0056] Reference memory error frequency = Number of reference memory errors / Total number of arm insertions;

[0057] Average exploration time = test time / total arm insertion time.

[0058] 3.2 Preparation of serum

[0059] After the last administration, 2 ml of blood was collected from each eye of each mouse in the eight-arm maze test group and placed in a test tube at 4°C overnight. The supernatant was collected the next day for use.

[0060] The levels of tryptophan, L-kynurenine, and 5-hydroxytryptamine in serum were detected by high performance liquid chromatography.

[0061] 4. Experimental Results:

[0062] 4.1 Results of the behavioral experiment

[0063] Table 1 Comparison of mean exploration time, working memory error frequency, and reference memory error frequency among mice in each group.

[0064]

[0065] Note: Compared to the model group + P<0.05, ++ P<0.01, +++ P<0.001;

[0066] Compared with the low-dose group of compound 1, & P<0.05, && P<0.01, &&& P<0.001;

[0067] Compared with the high-dose group of compound 1, ★ P<0.05, ★★ P<0.01, ★★★ P<0.001;

[0068] Compared with the low-dose icariin group, ＊ P<0.05, ＊＊ P<0.01, ＊＊＊ P<0.001;

[0069] Compared with the high-dose icariin group, ﹟ P<0.05, ﹟﹟ P<0.01, ﹟﹟﹟ P<0.001;

[0070] Compared with composition group A, △ P<0.05, △△ P<0.01, △△△ P<0.001;

[0071] Compared with composition group B, ◇ P<0.05, ◇◇ P<0.01, ◇◇◇ P<0.001;

[0072] Compared with composition group C, ○ P<0.05, ○○ P<0.01, ○○○ P<0.001.

[0073] Experimental results showed that the composition could significantly reduce the average exploration time, working memory error frequency and reference memory error frequency in AD model mice, and significantly improve cognitive dysfunction in mice. The effect was better than that of high and low dose groups of compound 1 and high and low dose groups of icariin.

[0074] Compared with compositions B and C, composition D showed better improvement in cognitive dysfunction in mice, while composition A showed the second best effect.

[0075] 4.2 Serum levels of tryptophan, L-kynurenine, and serotonin

[0076] Table 2 Comparison of serum 5-hydroxytryptamine, tryptophan, and L-kynurenine levels in mice of different groups

[0077]

[0078] Note: Compared to the model group + P<0.05, ++ P<0.01, +++ P<0.001;

[0079] Compared with the low-dose group of compound 1, & P<0.05, && P<0.01, &&& P<0.001;

[0080] Compared with the high-dose group of compound 1, ★ P<0.05, ★★ P<0.01, ★★★ P<0.001;

[0081] Compared with the low-dose icariin group, ＊ P<0.05, ＊＊ P<0.01, ＊＊＊ P<0.001;

[0082] Compared with the high-dose icariin group, ﹟ P<0.05, ﹟﹟ P<0.01, ﹟﹟﹟ P<0.001;

[0083] Compared with composition group A, △ P<0.05, △△ P<0.01, △△△ P<0.001;

[0084] Compared with composition group B, ◇ P<0.05, ◇◇ P<0.01, ◇◇◇ P<0.001;

[0085] Compared with composition group C, ○ P<0.05, ○○ P<0.01, ○○○ P<0.001.

[0086] Experimental results showed that the composition could significantly increase serum 5-hydroxytryptamine and tryptophan levels in AD model mice and significantly decrease serum L-kynurenine levels in AD model mice, with better effects than the high and low dose groups of compound 1 and the high and low dose groups of icariin.

[0087] Compared with compositions B and C, composition D showed better effects in increasing serum 5-hydroxytryptamine and tryptophan levels and decreasing L-kynurenine levels, while composition A showed the second best effect. The compositions of this invention can effectively regulate the synthesis of monoamine neurotransmitters, inhibit tryptophan metabolism in vivo, and improve learning and memory abilities.

Claims

1. A pharmaceutical composition for treating Alzheimer's disease, characterized in that, The pharmaceutical composition contains (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole and icariin, wherein the weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole to icariin is (0.5~10):

1.

2. The pharmaceutical composition according to claim 1, characterized in that, The weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole to icariin is (1.6~3.3):

1.

3. The pharmaceutical composition according to any one of claims 1-2, characterized in that, This pharmaceutical composition is a solid oral dosage form.

4. The pharmaceutical composition according to claim 3, characterized in that, This solid oral dosage form can be in the form of tablets, capsules, granules, or pills.

5. Use of the pharmaceutical composition of claim 1 in the preparation of a medicament for treating Alzheimer's disease.

6. The use as described in claim 5, characterized in that, The weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole to icariin is (1.6~3.3):

1.

7. The use as described in claim 5, characterized in that, The pharmaceutical composition described herein can increase serum 5-hydroxytryptamine and tryptophan levels, decrease serum L-kynurenine levels, and improve learning and memory abilities.

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