Application of pyroglutamic acid butyl ester in preparation of anti-Alzheimer disease medicine

Abstract

The invention discloses an application of pyroglutamic acid butyl ester in preparation of an anti-Alzheimer disease drug, and experiments prove that the pyroglutamic acid butyl ester has a remarkable improvement effect on memory and space exploration capability of an Alzheimer disease model mouse caused by A beta 25-35, and can enhance the activity of choline acetyltransferase in cortex and hippocampus; meanwhile, the traditional Chinese medicine composition also has a remarkable improvement effect on the memory and space exploration capability of model rats with Alzheimer's disease caused by D-galactose and AlCl3, and can enhance the activity of SOD enzyme and relieve the damage of free radicals to brain tissues. Therefore, the pyroglutamic acid butyl ester has a remarkable anti-Alzheimer disease effect, can be applied to preparation of the anti-Alzheimer disease medicine, and has a good market prospect in development of clinical anti-Alzheimer disease medicines.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to application of butyl pyroglutamate in the preparation of anti-Alzheimer's disease drugs. Background Art

[0002] Alzheimer's disease (AD) is a degenerative disease of the central nervous system with an unclear etiology. It usually starts in old age and has a slow course. Patients mainly show memory and cognitive dysfunction, which continues to develop and worsen, accompanied by neurological symptoms such as personality changes and language disorders. The reduction of acetylcholine synthesis, release and reuptake in the brain is an important cause of AD, and it is also often accompanied by major pathological changes such as β-amyloid protein deposition and Tau protein hyperphosphorylation. With the aging of the global population, the number of AD patients continues to increase. At present, the main anti-AD drugs are donepezil, rivastigmine, huperzine A, manno-oligosaccharide sodium (GV-971), etc., but these drugs can only relieve symptoms to a certain extent and cannot fundamentally prevent the progression of the disease. Therefore, the research and development of anti-Alzheimer's drugs is particularly urgent.

[0003] Fresh bamboo juice is the liquid that naturally drips from the fresh stems of Phyllostrachys glauca, P. nuda and several other plants of the same genus after being heated or roasted. It is sweet and bitter in nature, cold in nature, and enters the heart and stomach meridians. It has the effects of clearing away heat and resolving phlegm, calming the nerves and opening the orifices. It is often used clinically to treat lung heat, cough with excessive phlegm, asthma and chest tightness, tongue stiffness due to stroke, excessive phlegm, and phlegm-heat convulsions in children. [1] Studies have shown that fresh bamboo sap mainly contains flavonoids, phenylpropanoids, amino acids, phenolic acids, inorganic elements and their glycoside components. [1,2] .

[0004] In the study of the chemical composition and activity of fresh bamboo sap, the present invention isolated L-butyl pyroglutamate (Butyl5-oxoprolinate or butyl pyroglutamate), the molecular formula of which is C9H 15 NO3. Literature shows that this compound can enhance melanin production and tyrosinase activity and has the potential to treat melanoma and vitiligo skin diseases [3] ; It can also reduce the toxicity of the anti-tumor drug cisplatin to the kidneys [4] However, there is no report on other pharmacological activities of this component. Therefore, the present invention studies the pharmacodynamics of this compound on a variety of model animals and finds that this compound has a significant anti-Alzheimer's disease effect.

[0005] The compound can be isolated from a variety of Chinese herbal medicines such as fresh bamboo sap, silkworm, dandelion, northern bupleurum, mulberry, etc. [4-6]; At the same time, according to the literature method [7] , prepared by chemical synthesis; it can also be purchased directly from the market.

[0006] References:

[0007] [1] Feng Huimin, Hu Yunfei, Zeng Qianyao, et al. Research progress on chemical composition and quality control methods of fresh bamboo sap[J]. Chinese Journal of Traditional Chinese Medicine, 2021, 46(11): 2753-2759.

[0008] [2] Lin Kaipeng. Study on chemical composition and quality standard of fresh bamboo sap[D]. Nanchang University, 2020.08.

[0010] [3]Lee CL, Gao ZA, Jhan YL, et al. Tuliposide H–J and bioactive components from the bulb of Amana edulis[J]. Molecules, 2021,26:5907.

[0012] [4]Lee D,Yu JS,Lee SR,et al.Beneficial effects of bioactive compounds in mulberry fruits against cisplatin-induced nephrotoxicity[J].Int.J.Mol.Sci.,2018,19:1117.

[0014] [5] Huang Jin, Liu Yan, Pan Juan, et al. Study on the chemical composition of the n-butanol layer of the aerial part of Bupleurum chinense[J]. Journal of Traditional Chinese Medicine, 2020, 48(1): 38-41.

[0015] [6] Yan Mengqi, Wei Yishen, Huang Wanyi, et al. Study on the chemical composition of silkworm[J]. Journal of Guangxi Normal University (Natural Science Edition), 2023, 41(2): 124-130.

[0016] [7] Peng Xiaohai, Luo Jin, Zhong Min, et al. Synthesis and application of L-pyroglutamate [J]. Chemistry and Bioengineering, 2011, 28, 10: 79-80. Summary of the invention

[0017] The purpose of the present invention is to provide application of butyl pyroglutamate in preparing anti-Alzheimer's disease drugs.

[0018] The molecular formula of the butyl pyroglutamate of the present invention is C9H 15 NO3, its chemical structure is as follows:

[0019]

[0020] Its chemical name is Butyl 5-oxoprolinate or Butyl 5-oxopyrrolidine-2-carboxylate; it can be one of the chiral isomers L-butyl pyroglutamate or D-butyl pyroglutamate, or it can be the racemic DL-butyl pyroglutamate.

[0021] The butyl pyroglutamate of the present invention can be prepared by separation and purification from fresh bamboo sap, silkworm, dandelion, bupleurum chinense, mulberry and other traditional Chinese medicines, and can also be prepared by a synthetic method or obtained by commercial purchase.

[0022] The present invention treats Aβ 25～35 The Morris water maze training test of AD model mice and the determination of choline acetyltransferase activity in the cortex and hippocampus tissues proved that butyl pyroglutamate has a significant improvement effect on the memory and spatial exploration ability of AD model mice. Therefore, it can be used to prepare anti-AD drugs, and at a large dose, its effect is better than the positive control drug Noofukang.

[0023] The present invention uses Morris water maze training tests on AD model rats caused by D-galactose and AlCl3 and determination of the activity of SOD enzyme in their serum to prove that butyl pyroglutamate has a significant improvement effect on the memory and spatial exploration ability of AD model rats, can enhance the activity of SOD enzyme, and reduce the damage of free radicals to brain tissue. When the dosage is large, its effect is also better than that of the positive control drug Naofukang.

[0024] The present invention uses a classic AD animal modeling method to replicate AD model mice and rats, and gives the model animals different doses of butyl pyroglutamate intervention, which shows that butyl pyroglutamate has a significant anti-AD effect and can be used to prepare anti-AD drugs. In addition, it is also shown that when the dose of butyl pyroglutamate is large (40 mg / kg for rats and 50 mg / kg for mice), it is significantly better than the positive control drug Naofukang group in improving the memory of model mice and rats and enhancing the activity of SOD and ChAT enzymes, indicating its good market development prospects as a clinical anti-Alzheimer's disease drug. DETAILED DESCRIPTION

[0025] Example 1: Synthesis of D-pyroglutamate butyl ester

[0026] Add 25.8g (0.2mol) of D-pyroglutamic acid and 274mL (3.0mol) of butanol to a 500mL four-necked flask, stir to dissolve, cool to 0°C in an ice-salt bath, slowly drop 17.4mL (0.24mol) of freshly distilled thionyl chloride, and control the reaction temperature at 0-5°C during the dropwise addition. Continue stirring and reacting for 30min after the dropwise addition is completed, add triethylamine to adjust the pH value to 7-8, then remove the ice-salt bath and slowly heat to 30°C to react until the raw material spots on TLC disappear. Remove the solvent butanol, unreacted dichlorosulfuron and triethylamine in the system by vacuum distillation to obtain white crystals. Dissolve with 100mL of ethyl acetate, filter to remove triethylamine hydrochloride, add saturated brine to the filtrate, wash 3 times, separate, dry the ethyl acetate layer with anhydrous sodium sulfate, filter, and vacuum distill the filtrate to obtain a light yellow oily substance, i.e., 31.4g of D-pyroglutamic acid butyl ester, with a yield of 84.8% and a purity of 99.2%. HR ESI-MS m / z:186.1137[M+H] + ,208.0959[M+Na] + , molecular formula is C9H 15 NO3; 13 C NMR (150MHz, DMSO-d6))δ:180.6(C-5),172.6(-COO),65.6(C-1'),56.9(C-2),31.2(C-2'),30.5(C-4),25.6(C-3),19.7(C-3'),13.9(C-4').By comparing with the corresponding data of L-pyroglutamic acid butyl ester, it was identified as D-pyroglutamic acid n-butyl ester.

[0027] Example 2: Evaluation of the efficacy of compounds on Alzheimer's disease model mice

[0028] 2.1 Experimental animals and test drugs

[0029] Experimental animals: 96 SPF healthy male Kunming mice, weighing 26-30 g, were purchased from Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd.

[0030] Reagents: L-pyroglutamic acid, D-pyroglutamic acid, and L-pyroglutamic acid butyl ester were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; D-pyroglutamic acid butyl ester was prepared according to the method in Example 1; Aβ 25～35 purchased from Sigma; ChAT enzyme assay kit purchased from Nanjing Jiancheng Bioengineering Institute; Noofukang purchased from Hubei Huazhong Pharmaceutical Co., Ltd.

[0031] 2.2 Condensed Aβ 25～35 Preparation

[0032] 2 mg Aβ 25～35Dissolve in 2 mL of sterile saline to a concentration of 1 mmol / L. Seal and incubate in a 37°C cell culture incubator for 96 hours to convert it into condensed Aβ 25～35 , and place in a 4℃ refrigerator for later use.

[0033] 2.3 Preparation and administration of mouse AD model

[0034] After the mice were anesthetized with ether, their ovaries were removed. Ten days after the operation, the mice were randomly divided into control group, Aβ 25～35 The control group was injected with normal saline into the lateral ventricle of mice. The other groups were injected with 1mmol / L Aβ into the lateral ventricle of mice. 25～35 5μL was used to prepare the AD mouse model. 7 days after the model was prepared, the control group and Aβ 25～35 The mice in group (model group) were intraperitoneally injected with 5 mL / kg normal saline; the mice in group L-pyroglutamate, group D-pyroglutamate, and group I L-pyroglutamate butyl ester were intraperitoneally injected with 25 mg / kg of the corresponding drugs; the mice in group II L-pyroglutamate butyl ester and group II D-pyroglutamate butyl ester were intraperitoneally injected with 50 mg / kg of L-pyroglutamate butyl ester and D-pyroglutamate butyl ester, respectively; the mice in group NAOFUKANG were orally given 200 mg / kg of NAOFUKANG; the drugs were administered continuously for 14 days.

[0035] 2.4 Morris water maze training and testing

[0036] Water maze training began on the 8th day of drug administration for a total of 7 days. The water maze platform was placed in the 4th quadrant, and each mouse was tested twice a day. The quadrant where the platform was located was excluded, and mice were placed in the middle of the other 3 quadrants against the wall. A test was considered complete when the mouse was on the platform for 10 seconds and did not go into the water again; the mice that could not get on the platform were recorded for 90 seconds, and the mice were guided to the platform and stayed on the platform for 30 seconds to strengthen their memory. The swimming process of the mice was filmed and recorded for a total of 1 week. This was the learning process of the mice. One week after the learning process, the platform was removed and the spatial exploration ability of the mice was tested.

[0037] 2.5 Determination of choline acetyltransferase (ChAT) activity in the cortex and hippocampus of experimental mice

[0038] The mice were decapitated and the brains were removed. The cortex and hippocampus were peeled off on an ice bath, weighed, and quickly frozen in liquid nitrogen. Then, 50 mmol / L of pre-cooled potassium phosphate buffer was added and a 10% tissue homogenate was prepared on an ice bath. The ChAT enzyme assay kit was used to assay the activity of ChAT enzyme in the cortex and hippocampus of the experimental mice according to the instructions.

[0039] 2.6 Statistical analysis

[0040] The experimental results are SPSS13.0 software package was used for variance analysis, and t-test was used for comparison among groups.

[0041] 2.7 Results and Conclusions

[0042] Using Naofukang as a positive control, the effect of butyl pyroglutamate on the spatial exploration ability of experimental mice was determined. The results (Table 1) showed that intraperitoneal injection of 25 mg / kg and 50 mg / kg butyl pyroglutamate could significantly shorten the Aβ 25～35 The incubation period of Alzheimer's disease model mice increased, the number of transitions increased, and the time spent in the first quadrant was prolonged; compared with the model group, intraperitoneal injection of L-pyroglutamate butyl ester group I, group II and D-pyroglutamate butyl ester showed significant differences (P < 0.01); compared with the positive control Naofukang group, intraperitoneal injection of 50 mg / kg L-pyroglutamate butyl ester and D-pyroglutamate butyl ester showed significant differences (P < 0.01). At the same time, compared with the model group, there was no significant difference between the L-pyroglutamate group and the D-pyroglutamate group (P > 0.05); compared with the L-pyroglutamate group and the D-pyroglutamate group, all dose groups of L-pyroglutamate butyl ester and D-pyroglutamate butyl ester showed significant differences (P < 0.01). This shows that whether it is L-pyroglutamate butyl ester or D-pyroglutamate butyl ester, it has a significant effect on Aβ 25～35 The memory and behavior of Alzheimer's disease model mice were significantly improved, among which the improvement effect of intraperitoneal injection of 50 mg / kg L-butyl pyroglutamate and D-butyl pyroglutamate was significantly better than that of the positive control drug Noofukang; but there was no significant difference between the D-butyl pyroglutamate group and the L-butyl pyroglutamate group.

[0043] Table 1 Effects of butyl pyroglutamate on spatial exploration ability of experimental mice (n=12)

[0044]

[0045] Note: 25～35 Group (model group) comparison, * P<0.05, ** P<0.01; compared with the Naofukang group: # P<0.05, ## P<0.01; compared with L-pyroglutamate group and D-pyroglutamate group, P<0.05, P<0.01.

[0046] At the same time, the activity of choline acetyltransferase in the cortex and hippocampus of the above experimental mice was measured, and the results (Table 2) showed that: 25～35Compared with the model group, there were significant differences among the L-pyroglutamate butyl ester group I, L-pyroglutamate butyl ester group II and D-pyroglutamate butyl ester group (P < 0.01), indicating that pyroglutamate butyl ester can significantly enhance Aβ 25～35 The activity of choline acetyltransferase in the cortex and hippocampus of Alzheimer's disease model mice was reduced, thereby improving the learning and memory function of Alzheimer's disease; compared with the positive control Naofukang group, the L-pyroglutamate butyl ester group and the D-pyroglutamate butyl ester group at a dose of 50 mg / kg had significant differences (P < 0.01). At the same time, compared with the model group, there was no significant difference between L-pyroglutamate and D-pyroglutamate (P > 0.05); and compared with the L-pyroglutamate group and the D-pyroglutamate group, the L-pyroglutamate butyl ester group I, group II and D-pyroglutamate butyl ester group had significant differences (P < 0.01). This further shows that both L-pyroglutamate butyl ester and D-pyroglutamate butyl ester can significantly enhance Aβ 25～35 The activity of choline acetyltransferase in the cortex and hippocampus of Alzheimer's disease model mice was 25～35 The memory and behavior of Alzheimer's disease model mice were significantly improved, among which the improvement effect of intraperitoneal injection of 50 mg / kg L-butyl pyroglutamate and D-butyl pyroglutamate was significantly better than that of the positive control drug Naofukang.

[0047] Table 2 Effects of butyl pyroglutamate on choline acetyltransferase activity in mouse cortex and hippocampus (n=12)

[0048]

[0049] Note: 25～35 Group (model group) comparison, * P<0.05, ** P<0.01; compared with the Naofukang group: # P<0.05, ## P<0.01; compared with L-pyroglutamate group and D-pyroglutamate group, P<0.05, P<0.01.

[0050] In summary, the above analysis shows that butyl pyroglutamate has an 25～35 The resulting Alzheimer's disease model mice have significant anti-Alzheimer's disease effects and improved memory function, so they can be used to prepare anti-Alzheimer's disease drugs, and show that they have good application and development prospects in the preparation of anti-Alzheimer's disease drugs.

[0051] Example 3: Evaluation of the efficacy of the compound on Alzheimer's disease model rats

[0052] 3.1 Experimental animals and test drugs

[0053] Experimental animals: Aged healthy male Wistar rats, weighing 350-390 g, a total of 108 rats, were purchased from the Experimental Animal Science Center of Nanchang University.

[0054] Reagents: L-pyroglutamic acid, D-pyroglutamic acid, and L-pyroglutamate butyl ester were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; D-pyroglutamate butyl ester was prepared according to Experiment 1; DL-pyroglutamate butyl ester was obtained by mixing D- and L-pyroglutamate butyl esters in a 1:1 ratio; D-galactose and AlCl3 were purchased from Sinopharm Chemical Reagent Co., Ltd.; SOD enzyme kit was purchased from Nanjing Jiancheng Bioengineering Institute; and Noofukang was purchased from Hubei Huazhong Pharmaceutical Co., Ltd.

[0055] 3.2 Preparation and administration of compound Alzheimer's disease animal model

[0056] The experimental Wistar rats were randomly divided into control group, model group, Naofukang group, L-pyroglutamic acid group, D-pyroglutamic acid group, L-pyroglutamic acid butyl ester group Ⅰ, L-pyroglutamic acid butyl ester group Ⅱ, D-pyroglutamic acid butyl ester group, DL-pyroglutamic acid butyl ester group, 12 rats in each group. The model group was intraperitoneally administered with 60 mg / kg D-galactose (prepared with normal saline) and orally administered with 5 mg / kg AlCl3 (prepared with double distilled water); the Naofukang group, L-pyroglutamic acid group, D-pyroglutamic acid group, L-pyroglutamic acid butyl ester group Ⅰ, L-pyroglutamic acid butyl ester group Ⅱ, D-pyroglutamic acid butyl ester group, DL-pyroglutamic acid butyl ester group and other experimental drug groups were administered with the same dose of D-galactose and AlCl3 as the model group by the same administration route, and the drug doses shown in Table 3 were also intraperitoneally injected for 90 consecutive days.

[0057] 3.3 Morris water maze training and testing

[0058] The water maze training of the experimental rats began on the 83rd day of drug administration for a total of 7 days. One week after the learning process, the platform was removed and the spatial exploration ability of the rats was tested.

[0059] 3.4 Determination of SOD enzyme activity in serum of experimental rats

[0060] After the water maze test, 10 rats were selected from each group, anesthetized with chloral hydrate, and the chest was opened quickly to collect blood from the heart. The serum was collected after centrifugation at 3500 rpm for 10 min and stored at -70°C. According to the instructions of the kit, the activity of SOD enzyme in the serum of each group of experimental rats was determined by ultraviolet spectrophotometry.

[0061] 3.5 Statistical analysis

[0062] The experimental results are SPSS13.0 software package was used for variance analysis, and t-test was used for comparison among groups.

[0063] 3.6 Results and Conclusions

[0064] Using Naofukang as a positive control, the effect of butyl pyroglutamate on the spatial exploration ability of experimental rats was determined. The results (Table 3) showed that all dose groups of 20mg / kg and 40mg / kg butyl pyroglutamate injected intraperitoneally could shorten the latency period of Alzheimer's disease model rats induced by D-galactose and AlCl3, increase the number of platform crossings, and prolong the time spent in the first quadrant. Compared with the model group, L-butyl pyroglutamate groups I and II, D-butyl pyroglutamate group, and DL-butyl pyroglutamate group all had significant differences (P < 0.01); compared with the positive control Naofukang group, L-butyl pyroglutamate group II, D-butyl pyroglutamate group, and DL-butyl pyroglutamate group all had significant differences (P < 0.01). At the same time, compared with the model group, the L-pyroglutamate group (40 mg / kg) and the D-pyroglutamate group (40 mg / kg) also had significant differences (P < 0.01); but compared with the L-pyroglutamate group and the D-pyroglutamate group, the butyl pyroglutamate group, whether low-dose group (20 mg / kg) or high-dose group (40 mg / kg), had significant differences (P < 0.01). This shows that butyl pyroglutamate has a significant improvement effect on the memory and behavior of Alzheimer's disease model rats induced by D-galactose and AlCl3, and its improvement effect is significantly better than that of pyroglutamate, and the improvement effect of intraperitoneal injection of 40 mg / kg of L-pyroglutamate butyl ester, D-pyroglutamate butyl ester, and DL-pyroglutamate butyl ester is significantly better than the positive control drug Naofukang.

[0065] Table 3 Effects of butyl pyroglutamate on spatial exploration ability of experimental rats (n=12)

[0066]

[0067] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the Naofukang group: # P<0.05, ## P<0.01; compared with L-pyroglutamate group and D-pyroglutamate group, P<0.05, P<0.01.

[0068] At the same time, the activity of SOD enzyme in the serum of the above experimental rats was measured, and the results (Table 4) showed that compared with the model group, all the dose groups of intraperitoneal injection of 20mg / kg and 40mg / kg butyl pyroglutamate had significant differences (P<0.01), which showed that whether it was L-butyl pyroglutamate, D-butyl pyroglutamate or DL-butyl pyroglutamate, it could significantly enhance the activity of SOD enzyme in the serum of Alzheimer's disease model rats caused by D-galactose and AlCl3, reduce the damage of free radicals to brain tissue, and protect brain cells and tissues. Compared with the positive control group of Naofukang, the L-butyl pyroglutamate group, D-butyl pyroglutamate or DL-butyl pyroglutamate group at a dose of 40mg / kg had significant differences (P<0.01). At the same time, compared with the model group, the L-pyroglutamate group (40 mg / kg) and the D-pyroglutamate group (40 mg / kg) also had significant differences (P < 0.01), but compared with the L-pyroglutamate group and the D-pyroglutamate group, the butyl pyroglutamate group, whether low-dose group (20 mg / kg) or high-dose group (40 mg / kg), had significant differences (P < 0.01). This further shows that butyl pyroglutamate can significantly enhance the activity of SOD enzyme in the serum of Alzheimer's disease model rats caused by D-galactose and AlCl3, reduce the damage of free radicals to brain tissue, and protect brain cells and tissues. Its effect is significantly better than that of L- and D-pyroglutamate, and the effect of intraperitoneal injection of 40 mg / kg of L-pyroglutamate butyl ester, D-pyroglutamate butyl ester, and DL-pyroglutamate butyl ester is significantly better than the positive control drug Naofukang.

[0069] Table 4 Effect of butyl pyroglutamate on SOD activity in serum of experimental rats (n=12)

[0070]

[0071] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the Naofukang group: # P<0.05, ## P<0.01; compared with L-pyroglutamate group and D-pyroglutamate group, P<0.05, P<0.01.

[0072] The above description and analysis show that butyl pyroglutamate has a significant anti-Alzheimer's disease effect and improves memory function in Alzheimer's disease model rats induced by D-galactose and AlCl3, which further proves that butyl pyroglutamate can be used for the preparation of anti-Alzheimer's disease drugs, and also shows that it has good application and development prospects in the preparation of anti-Alzheimer's disease drugs.

[0073] Example 4: Preparation of D-pyroglutamate butyl ester capsules

[0074] (1) Prescription: 10 mL of D-pyroglutamate butyl ester, 12 g of starch, 8 g of lactose, appropriate amounts of sodium lauryl sulfate, 4% PVP, and magnesium stearate, a total of 200 capsules.

[0075] (2) Preparation: Dissolve D-pyroglutamate butyl ester in an appropriate amount of 95% ethanol, mix with starch, lactose and sodium dodecyl sulfate in a mixer and stir for 15 minutes. Slowly add 4% PVP as a binder while stirring, stir at high speed for 5 minutes, pass the wet granules through a 16-mesh sieve, dry in an oven at 55-60°C, and then granulate. Add lubricant magnesium stearate and mix well. Fill into capsules to prepare D-pyroglutamate butyl ester capsules.

[0076] Example 5: Preparation of L-pyroglutamate butyl ester tablets

[0077] (1) Prescription: 100 mL of L-pyroglutamate butyl ester, 20 g of starch, 12 g of 10% starch slurry, 11 g of dry starch, 2 g of magnesium stearate, made into 500 tablets.

[0078] (2) Preparation: Dissolve L-pyroglutamate butyl ester in an appropriate amount of 95% ethanol, mix with starch, add starch slurry to make a soft material, squeeze with a 14-mesh sieve to granulate, dry at 70-80°C, sieve with a 12-mesh sieve to size the granules, add dry starch and magnesium stearate, mix well, and then press into tablets to prepare L-pyroglutamate butyl ester tablets.

[0079] Therefore, the present invention proves through experiments that butyl pyroglutamate has an effect on Aβ 25～35 The memory and spatial exploration ability of Alzheimer's disease model mice caused by D-galactose and AlCl3 were significantly improved, and the activity of choline acetyltransferase in the cortex and hippocampus was enhanced. The effect of intraperitoneal injection of 50 mg / kg butyl pyroglutamate was significantly better than the daily dose of the positive control drug Naofukang (200 mg / kg). At the same time, it also significantly improved the memory and spatial exploration ability of Alzheimer's disease model rats caused by D-galactose and AlCl3, and enhanced the activity of SOD enzyme, reducing the damage of free radicals to brain tissue, and its effect was significantly better than pyroglutamic acid. The effect of intraperitoneal injection of 40 mg / kg butyl pyroglutamate was significantly better than the daily dose of the positive control drug Naofukang (150 mg / kg). It shows that butyl pyroglutamate has a significant anti-Alzheimer's disease effect, can be used to prepare anti-Alzheimer's disease drugs, and shows its good market prospect as a clinical anti-Alzheimer's disease drug development.

Claims

1. Application of butyl pyroglutamate in the preparation of anti-Alzheimer's disease drugs.

2. The use according to claim 1, characterized in that: The molecular formula of the butyl pyroglutamate is C9H 15 NO3, chemical structure is as follows: 。 3. The use according to claim 1, characterized in that: The butyl pyroglutamate is an isomer of the chiral isomers L-butyl pyroglutamate and D-butyl pyroglutamate, or a rotational isomer DL-butyl pyroglutamate.

4. The use according to claim 1, characterized in that: The drug dosage form is capsule, tablet or granule.

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