阅读说明：本技术 正丁醇基-2-O-（L-焦谷氨酸-N-6-）-α-D-呋喃果糖苷的新应用 (New application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside ) 是由 雷霞 李冰 雷双媛 张正一 张子涵 王来 于 2020-08-06 设计创作，主要内容包括：本发明公开了一种正丁醇基-2-O-(L-焦谷氨酸-N-6-)-α-D-呋喃果糖苷的新应用,包括在治疗阿尔茨海默病中的应用、在神经元细胞增殖中的应用、在学习记忆和认知能力改善中的应用、在抑制AchE活性、提高ChAT及Ach含量中的应用。本发明中的L-焦谷氨酸α的空间构型使其可与毒蕈碱型受体M3结合,L-焦谷氨酸α与毒蕈碱型受体M3结合后,使阿尔茨海默病患者大脑中的Ach保留下来,从而提高了Ach含量,而L-焦谷氨酸β则无法与毒蕈碱型受体M3结合,这也说明了L-焦谷氨酸α具有更加优异的抗AD作用,本发明通过分子生物学为L-焦谷氨酸α抗AD提供了有效性依据,从而为开发抗AD药的研究提供新思路。(The invention discloses a new application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside, which comprises an application in treating Alzheimer disease, an application in proliferating neuron cells, an application in improving learning memory and cognitive ability, and an application in inhibiting AchE activity and increasing ChAT and Ach contents. The spatial configuration of the L-pyroglutamic acid alpha in the invention enables the L-pyroglutamic acid alpha to be combined with a muscarinic receptor M3, after the L-pyroglutamic acid alpha is combined with a muscarinic receptor M3, the Ach in the brain of an Alzheimer disease patient is retained, so that the Ach content is improved, and the L-pyroglutamic acid beta cannot be combined with a muscarinic receptor M3, which also indicates that the L-pyroglutamic acid alpha has more excellent anti-AD effect.)

1. Application of N-butanol-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside in treating Alzheimer disease is provided.

2. Use according to claim 1, characterized in that: including use in neuronal cell proliferation.

3. Use according to claim 2, characterized in that: neuronal cells include PC12 cells.

4. Use according to claim 1, characterized in that: including use in learning memory and cognitive improvement.

5. Use according to claim 4, characterized in that: including the application in inhibiting the activity of AchE and increasing ChAT and Ach content.

6. Use according to claim 5, characterized in that: the N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside competitively binds with the muscarinic receptor M3 with Ach, and after the N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside binds with the muscarinic receptor M3, the Ach in the brain of the Alzheimer disease patient is retained, thereby relatively increasing the Ach content.

7. Use according to claim 1, characterized in that: including increased levels of ER α in the hypothalamic region; the application of the composition in reducing the content of ER beta, P-P38/P38 and APP and BACE1 proteins in the hippocampus.

8. Use according to claim 1, characterized in that: including the application in reducing the expression of apoptosis factors Bad and Caspase-3 and increasing the expression of anti-apoptosis factor Bcl-2.

9. Use according to claim 7, characterized in that: including the application of the ER-P38/MAPK signal channel in regulating the A beta metabolic system.

Technical Field

The invention relates to a new application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside, belonging to the technical field of pharmacy.

Background

Alzheimer's Disease (AD) is a progressive degenerative disease of the nervous system with occult onset. Clinically, the overall dementia such as dysmnesia, aphasia, disuse, agnosia, impairment of visual spatial skills, dysfunction in execution, and personality and behavior changes are characterized, and the etiology is unknown. Patients who are older than 65 years are called presenile dementia; the patient after 65 years old is called senile dementia.

At present, 5 anti-AD drugs are qualified in the market all over the world, namely donepezil, rivastigmine, galantamine, memantine and a donepezil and memantine compound preparation, and the drugs are all chemical drugs, only act on a single pathogenic mechanism to improve AD symptoms, and cannot delay the development of diseases.

In addition, the existing drugs are not deeply researched on the improvement mechanism of AD, and the therapeutic effect of the drugs cannot be verified from the perspective of molecular biology. The invention can provide a basis for the effectiveness of the N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside in resisting AD, and provides a new idea for the research of developing anti-AD drugs.

Disclosure of Invention

In order to solve the technical problems mentioned in the background technology, the invention provides a new application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

application of N-butanol-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside in treating Alzheimer disease is provided. Namely the application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside as a medicament for treating Alzheimer disease.

The use in neuronal cell proliferation. Namely the application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside as a neuron cell proliferation medicament.

Neuronal cells include PC12 cells.

The application is applied to the improvement of learning memory and cognitive ability. Namely the application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside as a learning and memory improving medicament and a cognition ability improving medicament.

The application comprises the application in inhibiting the activity of AchE and improving ChAT and Ach content.

The N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside competitively binds with the muscarinic receptor M3 with Ach, and after the N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside binds with the muscarinic receptor M3, the Ach in the brain of the Alzheimer disease patient is retained, thereby relatively increasing the Ach content.

The N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is used as an Ach competitive inhibitor.

The N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is applied as an AchE activity inhibitor.

The N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is applied as a ChAT content increasing agent.

The application increases ER alpha content in hypothalamus regions; the application of the composition in reducing the content of ER beta, P-P38/P38 and APP and BACE1 proteins in the hippocampus.

The N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is used as an ER alpha content increasing agent, an ER beta content reducing agent, a P-P38/P38 content reducing agent, an APP protein content reducing agent and a BACE1 protein content reducing agent.

The application is the application in reducing the expression of apoptosis factors Bad and Caspase-3 and increasing the expression of anti-apoptosis factors Bcl-2.

The N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is used as an apoptosis factor inhibitor and an anti-apoptosis factor promoter, wherein the apoptosis factors comprise Bad and Caspase-3, and the anti-apoptosis factors comprise Bcl-2.

The application is applied to the ER-P38/MAPK signal pathway to regulate the A beta metabolic system.

The N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is used as an ER-P38/MAPK signal pathway regulator.

the N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) - α -D-fructofuranoside in the invention is white powder, ESI-MS⁺[ M + H ] is visible at M/z348]⁺Ion peak, indicating the molecular weight of the compound is 347, determining the molecular formula as C₁₅H₂₅NO₈Acid hydrolysis of the acid gave D-fructose, suggesting that the compound contained fructose.

¹Hydrogen proton signals ascribed to fructose were observed in 4.35(1H, d,11.6, H-1-a), 4.24(1H, d,11.6, H-1-b), 3.96(1H, d,8.0, H-3), 3.91(1H, d,8.0, H-4) in H-NMR; 3.74(1H, m, -OCH)₂-a)，3.52(1H,m，-OCH₂-b)，1.53(2H,m,-CH₂)，1.37(2H,m,-CH₂)，0.92(3H,t,7.2,-CH₃) Hydrogen proton signals attributed to n-butanol groups; 4.32(1H, dd,8.0,4.0, -CH), 2.49(1H, m, -CH) were also observed₂-a)，2.33(2H,m,-CH₂)，2.23(1H,m,-CH₂-b) hydrogen proton signal attributed to pyroglutamic acid.

In the spectrum¹³In C-NMR, a total of 15 carbon signals were observed, with the remaining 5 carbon signals 181.2, 173.4, 57.1, 30.2, 25.8 being attributed to L-pyroglutamic acid, in addition to the 6 carbon signals attributed to fructose and 4 carbon signals attributed to n-butyl.

The methine proton signal 4.32(1H, dd,8.0,4.0, -CH) on glutamate observed in the HMBC spectrum was also correlated with the carbonyl carbon signal (173.4) in addition to the free carboxyl carbon signal (181.2), indicating that glutamate forms L-pyroglutamic acid; a hydrogen proton signal attributable to n-butanol hydroxymethyl was observed as 3.74(1H, m, -OCH)₂-a) correlates with the fructose C-2 carbon signal (102.9), indicating that n-butyl is linked at the 2-position of fructose; it was also observed that both hydrogen proton signals 4.35(1H, d,11.6, H-1-a), 4.24(1H, d,11.6, H-1-b) of the hydroxymethyl group at position 1 on fructose were related to the lactonamide carbonyl carbon signal (173.4) of pyroglutamic acid, indicating that L-pyroglutamic acid is glycosidically bonded to the hydroxymethyl group at position 1 of fructose via a nitrogen atom.

In conclusion, combining DEPT and HSQC spectra¹H-NMR and¹³C-NMR data are assigned, and the structure of the compound is identified as N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) - α -D-fructofuranoside, which is abbreviated as L-pyroglutamic acid α.

The isomer of N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -beta-D-fructofuranoside.

N-butanol-2-O- (L-pyroglutamic acid-N-6-) - β -D-fructofuranoside is white powder, ESI-MS⁺[ M + H ] is visible at M/z348]⁺Ion peak, indicating that the molecular weight of the compound should be 347, determining the molecular formula as C₁₅H₂₅NO₈The unsaturation was calculated to be 4. Acid hydrolysis gave D-fructose, suggesting that this compound contained fructose.

¹The hydrogen proton signals ascribed to fructose were observed in 4.28(2H, o, H-6), 4.12(1H, d,8.0, H-3), 3.99(1H, d,8.0, H-4), 3.91(1H, m, H-5) by H-NMR; 3.61(1H, m, -OCH)₂-a),3.45(1H,m，-OCH₂-b)，1.53(2H,m,-CH₂)，1.37(2H,m,-CH₂)，0.91(3H,t,7.2,-CH₃) Hydrogen proton signals attributed to n-butanol groups; 4.30(1H, m, -CH-5), 2.49(1H, m, -CH)₂-a-3)，2.34(2H,m,-CH₂-4)，2.21(1H,m,-CH₂-b-3) hydrogen proton signal attributed to pyroglutamic acid.

In the spectrum¹³in C-NMR, of the 15 carbon signals, 4 carbon signals ascribed to n-butyl and 5 carbon signals 181.2, 173.8, 57.0, 30.2 and 25.9 carbon signals ascribed to L-pyroglutamic acid, the remainder being fructose were compared to find that the resulting product was β -D-fructofuranose, and the 6 hydroxymethyl carbon was shifted to the low field by 2.5ppm in addition to the 2 carbon.

The methine proton signal 4.30(1H, m, -CH-5) on glutamate observed in the HMBC spectrum was correlated with the carbonyl carbon signal (173.4) in addition to the free carboxyl carbon signal (181.2), further confirming the formation of L-pyroglutamic acid; a hydrogen proton signal attributable to n-butanol hydroxymethyl was observed as 3.61(1H, m, -OCH)₂-a) correlation with the fructose C-2 carbon signal (105.6)Indicating that n-butyl is attached at the 2-position of fructose; two hydrogen proton signals 4.28(2H, o, H-6) for the 6-hydroxymethyl group on fructose were observed to correlate with the lactonamide carbonyl carbon signal of pyroglutamic acid (173.8), indicating that L-pyroglutamic acid is glycosidic to the 1-hydroxymethyl group of fructose via the nitrogen atom.

Combined DEPT and HSQC spectra¹H-NMR and¹³C-NMR data are assigned, and the structure of the compound is identified as N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) - β -D-fructofuranoside, which is abbreviated as L-pyroglutamic acid β.

L-pyroglutamic acid alpha and L-pyroglutamic acid beta are collectively referred to as L-pyroglutamic acid in the present invention.

The invention has the beneficial effects that: compared with the blank group, the scopolamine-induced mice have obviously reduced learning and memory abilities, the P-P38/P38, APP and BACE1 are obviously increased (P is less than 0.01), the number of neurons in hippocampal and hypothalamus areas is reduced, mitochondria are flocculent and denatured, rough endoplasmic reticulum is expanded and degranulated, nism of Nissn bodies disappears, the structure of synaptic membranes is unclear, and the number of vesicles is reduced. Compared with a model group, the L-pyroglutamic acid alpha can obviously increase the new object recognition index, the escape latency, the platform crossing times and the target quadrant residence time (P <0.01) of a model mouse, reduce the expression (P <0.01) of P-P38/P38, APP and BACE1, and ensure that the L-pyroglutamic acid alpha administration group has the advantages of compact arrangement, more number, normal shape, obviously increased vesicle number, clear cell double-layer nuclear membrane, obvious nucleolus and developed mitochondria. Therefore, L-pyroglutamic acid alpha can regulate A beta metabolic system to achieve neuroprotective effect on scopolamine dysmnesia model mice through ER-P38/MAPK signal channel.

In addition, the spatial configuration of the L-pyroglutamic acid alpha in the invention leads the L-pyroglutamic acid alpha to be combined with the muscarinic receptor M3, and after the L-pyroglutamic acid alpha is combined with the muscarinic receptor M3, the Ach in the brain of the Alzheimer disease patient is reserved, so that the content of the Ach is relatively increased. The invention verifies whether the L-pyroglutamic acid alpha passes through ER-P38/MAPK signals through molecular biology, regulates A beta metabolism, generates a protective effect on hippocampal neurons, provides an effective basis for resisting AD by the L-pyroglutamic acid alpha, and thus provides a new idea for the research of anti-AD drugs.

Drawings

FIG. 1 is a structural formula diagram of N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside in the present invention;

FIG. 2 is a structural formula diagram of N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -beta-D-fructofuranoside in the present invention;

FIG. 3 is a graph of cell proliferation rate of the present invention;

FIG. 4 is a graph showing the effect of L-pyroglutamic acid on the expression of ER β, P-P38/P38, APP, BACE1 in mouse hippocampus;

FIG. 5 is a three-dimensional view of the molecular docking of L-pyroglutamic acid alpha with the M3 receptor of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.

The new application of N-butyl alcohol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside specifically comprises the following contents.

As shown in FIG. 1, FIG. 1 is the structural formula of N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside, wherein the large groups on C1 and C4 of fructose in N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside are positioned at two sides, namely the glycosidic bond is alpha type, namely N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -alpha-D-fructofuranoside is abbreviated as L-pyroglutamic acid alpha.

As shown in FIG. 2, FIG. 2 is the structural formula of N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -beta-D-fructofuranoside, wherein the large groups on C1 and C4 of fructose in N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -beta-D-fructofuranoside are located on the same side, i.e. the glycosidic bond is beta, i.e. N-butanol group-2-O- (L-pyroglutamic acid-N-6-) -beta-D-fructofuranoside is abbreviated as L-pyroglutamic acid beta.

L-pyroglutamic acid α and L-pyroglutamic acid β in the present example are collectively referred to as L-pyroglutamic acid.

first, as shown in FIG. 3, L-pyroglutamic acid has a good protective effect on A β induced PC12 cell damage, and L-pyroglutamic acid significantly increases the proliferation of damaged PC12 cells and increases the survival rate of neurons, wherein the model group is A β_25-35Injury model, positive drug group was donepezil.

And secondly, the L-pyroglutamic acid has the function of improving the learning memory and cognitive ability of the scopolamine memory impairment model mouse.

Grouping and administration, 75 mice were randomly divided into 5 groups, a blank group (Control), a model group (Scopolamine), a positive drug group (Donepezil), an L-pyroglutamic acid alpha group, and an L-pyroglutamic acid beta group. Feeding distilled water into the stomach of the Control group and the Scopolamine group; donepezil (585 mg/kg. d) and L-pyroglutamic acid (100 mg/kg. d) were administered to the stomach of the Donepezil group and the Scopolamine group, respectively, and after 15min, L-pyroglutamic acid (100 mg/kg. d) was administered. On the 22 th day of molding, except for the blank group, the same amount of scopolamine (100 mg/kg. d) was intraperitoneally injected, and the behavioral tests were started on 7 consecutive days and 29 th day, which were performed 30min after daily administration.

The behavioral determination adopts a new object recognition experiment (ORT) and a Morris water maze experiment (MWM) to observe the cognitive ability of the mice. Recognition index ═ Tn-Tf)/(Tn + Tf). The contact Time to the new object is (Time new) and the contact Time to the old object is (Time family); the Morris water maze experiment comprises a positioning navigation experiment and a space exploration experiment. The number of times the animals passed through the platform, with latency to escape, was recorded.

Pathological methods detect hippocampal and hypothalamic morphology. HE staining: the hippocampus and hypothalamus were removed and fixed with 40% paraformaldehyde for 48h, embedded in paraffin and sectioned. Pathological changes of the mouse hippocampus and hypothalamus were observed under a light microscope. Fixing by a transmission electron microscope, dehydrating, soaking, embedding and slicing, observing the change of the ultrastructure of the mouse hippocampus under the electron microscope and shooting. And (4) dripping primary antibody and secondary antibody diluted in a proper proportion according to a method of a reagent instruction, and observing ER expression. The calculation is carried out by adopting a Motic Med 6.0 digital medical image analysis system.

After the behavior test of related protein expression in mouse hippocampus, 1% sodium pentobarbital (0.1mL/10g) is given to the mouse for intraperitoneal injection and anesthesia, the head of each group of mice is rapidly cut off after anesthesia, hippocampus and hypothalamus in mouse brain tissues are taken out, the whole operation process is carried out on ice, protein is extracted after liquid nitrogen quick freezing, the protein concentration is measured according to the operation of a BCA kit specification, the protein concentration of each group of samples is calculated, and the protein expression level is detected by Western blot.

Statistical analysis data were analyzed using SPSS21.0 statistical software, and comparisons between groups were performed using one-way ANOVA, experimental data were expressed as mean. + -. standard deviation (x. + -. SD), and P <0.05 was statistically significant.

Results

The results of the new object identification experiment are shown in table 1, and compared with the blank group, the identification index of the new object of the Scopolamine group mouse is obviously reduced (P < 0.01); the index of novel object recognition was significantly increased in mice of the Donepezil and L-pyroglutamate α group compared to the Scopolamine group (P < 0.01); compared with the L-pyroglutamic acid alpha group, the mouse of the L-pyroglutamic acid beta group has a remarkably reduced index of new object recognition (P < 0.01).

TABLE 1 Effect of L-Pyroglutamic acid on mouse neobody recognition index ((S))

n＝15)

Note: p <0.01 compared to Control group; in contrast to the Scopolamine group, # # indicates P < 0.01; and & means P <0.01, compared with the L-pyroglutamic acid α group.

Morris water maze test results: positioning navigation experiment: the results are shown in Table 2, with a significant increase in latency (P <0.01) in the Scopolamine group compared to the blank group and a significant decrease in latency (P <0.01) in the Donepezil and L-pyroglutamate alpha group compared to the Scopolamine model group; latency was significantly reduced in L-pyroglutamic acid β group mice compared to L-pyroglutamic acid α group (P < 0.01). Meanwhile, the length of the platform latency period searched in the first 4d training of the 5 groups of mice is recorded, and it can be seen that the latency periods of the 5 groups are gradually reduced, but the latency periods of the Control group and the L-pyroglutamic acid alpha group of mice are reduced more quickly, the latency periods of the Scopolamine group and the L-pyroglutamic acid beta group are shortened less, and the progress is slow, so that the learning ability of the mice is poor.

TABLE 2 Effect of L-pyroglutamic acid on mouse localization navigation latency: (

n＝15)

Note: p <0.01 compared to Control group; in contrast to the Scopolamine group, # # indicates P < 0.01; and & means P <0.01, compared with the L-pyroglutamic acid α group.

As can be seen from table 3, the number of platform crossings and target quadrant residence time was significantly reduced in the Scopolamine group mice compared to Control (P < 0.01); the number of platform crossings and target quadrant residence times were significantly increased in mice of the Donepezil and L-pyroglutamate α groups compared to the Scopolamine group (P < 0.01); compared with the L-pyroglutamic acid alpha group, the platform crossing times and the target quadrant residence time of the mice in the L-pyroglutamic acid beta group are remarkably reduced (P <0.01), and the target quadrant residence time is reduced (P < 0.05).

TABLE 3 influence of L-pyroglutamic acid on the number of mouse space exploration trial passes and target quadrant residence time: (

n＝15)

Note: p <0.01 compared to Control group; in contrast to the Scopolamine group, # # indicates P < 0.01; and & means P <0.01, compared with the L-pyroglutamic acid α group.

Influence of L-pyroglutamic acid on pathological changes of hippocampus and hypothalamus tissues of scopolamine model mouse

The hippocampal nerve cells of the mice in the Control group are regularly arranged, have a large number and are uniformly stained. Scopolamine damages mouse hippocampus, presents in disorderly arranged and reduced neuronal cells, and no obvious nucleolus can be seen due to obvious phenomenon of nuclear pyknosis; both the Donepzil group and the L-pyroglutamic acid alpha administration group recover the number of the neurons in the hippocampus to a certain degree, and the hippocampus has normal shape, regular arrangement and clear nucleolus. The HE staining of the hypothalamus also shows similar characteristics, the mouse hypothalamus is damaged by the scopolamine, the arrangement of nerve cells is loose, the intercellular space is enlarged, the staining of cell nucleus is light, the volume of the cell nucleus is increased, nucleolus is not obvious, part of the cell structure disappears, the nerve cells of the Donepzil and L-pyroglutamic acid alpha administration group are arranged more tightly, the intercellular space is enlarged, the staining of the cell nucleus is deeper, the nucleolus is clearer, and part of the cell structure is degenerated.

As can be seen from table 4, ER β expression was significantly reduced in the Scopolamine group mice compared to the Control group (P < 0.01); ER beta expression of mice in the L-pyroglutamic acid alpha group is obviously increased compared with that in the Scopolamine group (P < 0.01); ER beta expression was significantly reduced in mice in the L-pyroglutamic acid beta group compared to the L-pyroglutamic acid alpha group (P < 0.01). Compared with the Control group, the mice in the Scopolamine group have obviously reduced ER alpha expression in the hypothalamus region (P < 0.01); compared with the Scopolamine group, the ER alpha expression of mice in the administration group is obviously increased (P < 0.01); ER α expression was significantly reduced in mice in the L-pyroglutamic acid β group compared to the L-pyroglutamic acid α group (P < 0.01).

TABLE 4 Effect of L-pyroglutamic acid on mouse hippocampal P-P38/P38 expression ((S))

n＝3)

Note: p <0.01 compared to Control; , # # P <0.01 compared to the Scopolamine group; and & P <0.01 compared to the L-pyroglutamic acid alpha group.

As shown in fig. 4 and table 5, the content of APP and BACE1 in hippocampus was changed, and compared with Control group, APP of Scopolamine group and BACE1 expression were significantly increased (P <0.01), and compared with Scopolamine group, APP of administration group and BACE1 expression were significantly decreased (P < 0.01); compared with the L-pyroglutamic acid alpha group, the expression of APP and BACE1 in the hippocampus of the L-pyroglutamic acid beta group mice is obviously increased (P < 0.01).

In FIG. 4, A-Control group; B-Scopolamine group; C-Donepzil group; group D-L-pyroglutamic acid α; D-L-pyroglutamic acid beta group.

TABLE 5 Effect of NG on APP, BACE1 expression in the hippocampus of SN mice ((

n＝3)

Note: denotes P <0.01 compared to blank group; in contrast to the SN group, # # indicates P < 0.01; compared to the NG group, & & means P < 0.01.

AD is characterized pathologically by aberrant deposition of a β forming senile plaques and neurofibrillary tangles formed by hyperphosphorylation of Tau protein. BACE1(β -secretase) is the rate-limiting enzyme required by APP to produce A β. Neurofibrillary tangles formed by hyperphosphorylation of Tau protein are closely related to the degree of dementia in AD patients, and Cdk5 is one of cyclin-dependent kinases and is involved in early generation of neurofibrillary tangles in AD patients. The experimental result shows that L-pyroglutamic acid can reduce the expression of APP and BACE1, reduce the formation of neurofibrillary tangles and play a role in neuroprotection of the PC12 cells with A beta injury and scopolamine model mice.

The cholinergic system is related to learning and memory, and the AD patients have different degrees of Ach synthesis reduction in brain, wherein the Ach is one of important neurotransmitters of the cholinergic system, and the reduction of the Ach synthesis causes damage and loss of neurons and also causes synaptic signal transduction disorder. The experimental result shows that the L-pyroglutamic acid can inhibit the activity of AchE and improve the contents of ChAT and Ach, thereby playing a role in learning and memory protection of scopolamine model mice.

As shown in fig. 5, the spatial configuration of L-pyroglutamic acid alpha results in the binding to muscarinic receptor M3, and the binding of L-pyroglutamic acid alpha to muscarinic receptor M3 results in the retention of Ach in the brain of Alzheimer's patients, thereby relatively increasing the content of Ach. While L-pyroglutamic acid beta cannot be bound with muscarinic receptor M3, which also indicates that L-pyroglutamic acid alpha has more excellent anti-AD effect.

Neuronal apoptosis is an important physiological function in the central nervous system. Abnormal deposition of Abeta can induce neuron apoptosis in various ways, initiate cascade reaction of various proteins in neuron cells participating in apoptosis signals, induce oxidative stress and inflammatory reaction, and finally cause neuron apoptosis and loss to cause AD. The research finds that Bad, Bcl-2 and Caspase-3 belong to apoptosis-related proteins, normally, the apoptosis inhibiting protein and the apoptosis promoting protein are in a relative balance state, when Bcl-2 excessively inhibits the neuronal apoptosis, and Bad and Caspase-3 excessively promotes the neuronal apoptosis. The experimental result shows that the L-pyroglutamic acid can reduce the expression of apoptosis factors Bad and Caspase-3, increase the expression of anti-apoptosis factor Bcl-2, reduce the apoptosis of neurons and play a role in protecting the nerves of a scopolamine model mouse.

The L-pyroglutamic acid alpha of the embodiment can obviously improve the learning and memory functions of the memory impairment model mouse. The A beta metabolic system is improved through MAPK signals, and the neuroprotective effect on memory impairment model mice is exerted.

The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.