CN109096350B - New thioether compounds in Moringa seeds and their applications - Google Patents

Abstract

The invention relates to 4 new disulfide and trithioether compounds extracted and separated from moringa seeds and application of the disulfide compounds in preparing medicaments for treating diabetes, depression and senile dementia, and experiments show that the compounds have better effects of reducing blood sugar, depression and senile dementia. The thioether compound has the characteristics of clear activity mechanism, low toxicity, safety and the like, and has wide application prospect.

Description

New thioether compounds in Moringa seeds and their applications

technical field

The invention relates to new thioether compounds separated from Moringa seeds and their application in preparing medicines for treating diabetes, anti-depression and anti-senile dementia.

Background information

Moringa (Moringaoleifera Lam.) belongs to the genus Moringa, is a perennial tropical deciduous tree, native to tropical and subtropical arid and semi-arid regions, and is now widely distributed in tropical marine climates such as Africa, Arabia, Southeast Asia, and Pacific islands. District (Anwar F, et al. Phytother Res, 2007, 21(1):17-25). After the 1960s, it has been cultivated on a large scale in Yunnan, Hainan, Guangdong, Guangxi and other places in China (Dong Xiaoying et al. Guangdong Feed, 2008, 17(9): 39-41). Moringa is a multi-purpose fast-growing tree that can bloom and bear fruit after 6 months of planting. Its roots, stems, leaves, flowers and seeds can be used medicinally (Chinese Flora, 1984:34(1):6). The main chemical components in Moringa are phenols and their glycosides, flavonoids and their glycosides, sterols and their glycosides, as well as polysaccharides, amino acids and vitamins. Modern pharmacological studies have shown that Moringa oleifera has activities such as lowering blood sugar, lowering blood lipids, protecting liver damage, protecting nervous system, anti-tumor, anti-inflammatory, bacteriostatic, analgesic and antispasmodic (Kong Lingyu et al. Tianjin Pharmacy, 2015, 27(2):57 -59; Xu Min et al. Food Science, 2016, 37(23):291-301). However, the above pharmacological activities are all studies on the water or alcohol extracts of Moringa oleifera, and there is no report on the relevant activities of the monomer components.

In the present invention, a series of thioether compounds are found from Moringa seeds, including 5 new compounds, and it is found for the first time through research that these thioether compounds have the functions of treating diabetes, anti-depression and anti-senile dementia.

SUMMARY OF THE INVENTION

The invention relates to a new thioether compound separated from Moringa seeds and its application in preparing medicines for treating diabetes, anti-depression and anti-senile dementia.

The general chemical formula of the thioether compounds of the present invention and their derivatives is as follows:

wherein R ₁ and R ₂ are hydrogen or sugar group; n=1, 2 or 3, ether group is preferably monosulfide, disulfide, trisulfide, etc.; sugar group is preferably six carbons (glucose, mannose, rhamnose) ) pyranose, six-carbon furanose, and the acylated sugar is preferably six-carbon (glucose, mannose, rhamnose) pyranose, six-carbon furanose. or acyl sugars (acetyl, propionyl, benzoyl). Preferred compound substituents and names according to the present invention are shown in Table 1.

Table 1 Substituents and names of thioether compounds

When the compound of the present invention is used as a medicine, it can be used directly or in the form of a pharmaceutical composition. At least one compound of formula (I) is included as an active ingredient, in combination with one or more pharmaceutically acceptable carriers and excipients that are nontoxic and inert to humans.

The carriers and excipients used are one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical preparation auxiliaries. The pharmaceutical composition of the present invention is prepared into various dosage forms by methods recognized in the pharmaceutical field, such as liquid preparations (suspension, syrup, oral liquid or injection, etc.), solid preparations (tablets, capsules or granules, etc.) ), spray, etc. The above-mentioned drugs can be administered orally, sublingually or by injection (intravenous injection, intramuscular injection or subcutaneous injection, etc.) and other routes.

Description of drawings:

Fig. 1 is the ESI-MS picture of compound 1;

Fig. 2 is the hydrogen nuclear magnetic resonance spectrum of compound 1;

Fig. 3 is the carbon nuclear magnetic resonance spectrum of compound 1;

Fig. 4 is the ESI-MS picture of compound 2;

Fig. 5 is the hydrogen nuclear magnetic resonance spectrum of compound 2;

Fig. 6 is the carbon nuclear magnetic resonance spectrum of compound 2;

Fig. 7 is the ESI-MS picture of compound 3;

Fig. 8 is the hydrogen nuclear magnetic resonance spectrum of compound 3;

Fig. 9 is the carbon nuclear magnetic resonance spectrum of compound 3;

Figure 10 is the ESI-MS diagram of compound 4;

Figure 11 is the hydrogen nuclear magnetic resonance spectrum of compound 4;

FIG. 12 is the carbon nuclear magnetic resonance spectrum of compound 4. FIG.

Detailed ways

The present invention will be better understood by reference to the following examples, which are given to illustrate the invention, but not to limit the scope of the invention.

Example 1 Extraction, separation and structural identification of new compounds

Take 5 kg of Moringa seeds and pulverize, add 10 times the weight of 55% ethanol for diafiltration and extraction twice, 24h each time, combine the extracts, concentrate, filter, adsorb on the filtrate on a D101 macroporous resin column, and sequentially remove it by elution with water and 10% ethanol The impurities are eluted with 50 L of 80% ethanol, the 80% ethanol eluent is recovered, concentrated, and dried under reduced pressure to obtain the total extract. 150 g of the total extract was subjected to silica gel column chromatography, eluted with a petroleum ether-ethyl acetate system (volume ratio of petroleum ether and ethyl acetate from 100:0 to 1:1) gradient elution, and each gradient was recovered after solvent was combined to obtain fraction Fr .1-Fr.10. Fr.2 (28g) was subjected to ODS (reverse octadecyl bonded silica gel) column chromatography, methanol-water system gradient elution (methanol to water volume ratio from 2:8 to 1:0) to obtain 6 fractions, Fr.3a-Fr.3f, wherein Fr.3c (1.3 g) was subjected to preparative HPLC with 30% methanol-water as mobile phase to obtain compound 3 (535 mg). Fr.24 (18g) was subjected to ODS (reverse octadecyl bonded silica gel) column chromatography, acetonitrile-water system gradient elution (methanol to water volume ratio from 1:9 to 1:0) to obtain 5 fractions, Fr.3a-Fr.3e, wherein Fr.3b (1.5 g) was subjected to preparative HPLC with 35% methanol-water as mobile phase to obtain compound 4 (615 mg). Fr.3d (1.1 g) was subjected to preparative HPLC with 48% methanol-water as mobile phase to obtain compound 1 (465 mg). Fr.3e (2.1 g) was subjected to preparative HPLC with 40% acetonitrile-water as mobile phase to obtain compound 2 (442 mg).

Compound 1 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 569.1531 [MH] ⁻ . ESI-MS is shown in Figure 1. ¹ H-NMR (CD ₃ OD) δ: 7.18 (2H, d, J=8.5 Hz, H-2 and H-6), 7.02 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.42(1H,d,J=1.5Hz,H-1'), 3.99(1H,m,H-2'), 3.83(1H,dd,J=9.3,3.5Hz,H-3'), 3.60 (1H,m,H-5'), 3.56(2H,d,J=5.9Hz,H-7), 3.45(1H,t,J=9.3Hz,H-4'), 1.18(3H,d, J=6.2 Hz, H-6'). The H NMR spectrum is shown in Figure 2. ¹³ C-NMR (CD ₃ OD) δ: 157.2 (C-4), 132.6 (C-1), 131.7 (C-2 and C-6), 117.4 (C-3 and C-5), 99.7 (C -1'), 73.8(C-4'), 72.2(C-2'), 72.0(C-3'), 70.6(C-5'), 43.3(C-7), 18.0(C-6') ). The carbon NMR spectrum is shown in Figure 3. The above data indicate that compound 1 is a highly symmetrical phenolic glycoside compound containing disulfide.

Compound 1 was acid hydrolyzed to obtain aglycone and sugar moieties, and the sugar was subjected to derivatization reaction. Through HPLC liquid phase analysis and comparison with the derivatives of standard sugar, the sugar was identified as L-rhamnose, and the C-3— ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 1 is identified as S,S'-bis[(4-O-α-L-rhamnosyl)benzyl]-disulfide, which is a new compound.

Compound 2 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 601.1252 [MH] ⁻ . ESI-MS is shown in Figure 4. ¹ H-NMR (CD ₃ OD) δ: 7.24 (2H, d, J=8.5 Hz, H-2 and H-6), 7.02 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.41(1H,d,J=1.5Hz,H-1'), 3.98(1H,m,H-2'), 3.97(2H,d,J=5.9Hz,H-7), 3.83(1H, dd, J=9.3, 3.5Hz, H-3'), 3.61 (1H, m, H-5'), 3.44 (1H, t, J=9.3Hz, H-4'), 1.20 (3H, d, J=6.2 Hz, H-6'). The H NMR spectrum is shown in Figure 5. ¹³ C-NMR (CD ₃ OD) δ: 157.3 (C-4), 131.7 (C-2 and C-6), 117.5 (C-3 and C-5), 99.8 (C-1'), 73.8 ( C-4'), 72.2 (C-2'), 72.0 (C-3'), 70.6 (C-5'), 43.3 (C-7), 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 6. The above data indicate that compound 2 is a highly symmetrical phenolic glycoside compound containing trisulfide.

Compound 2 was acid hydrolyzed to obtain aglycone and sugar moiety, and the sugar was subjected to derivatization reaction. The HPLC liquid phase analysis was compared with the derivative of standard sugar, and the sugar was identified as L-rhamnose, combined with the C-3- of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 2 was identified as S,S″-bis[(4-O-α-L-rhamnosyl)benzyl]-trisulfide, which was a new compound.

Compound 3 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 423.0904 [MH] ⁻ . ESI-MS is shown in Figure 7. ¹ H-NMR (CD ₃ OD) δ: 7.19 (2H, d, J=8.5 Hz, H-2 and H-6), 7.06 (2H, d, J=8.5 Hz, H-2' and H-6 '), 7.02 (1H,d, J=8.5Hz, H-3 and H-5), 6.72 (1H,d, J=8.5Hz, H-3' and H-5'), 5.42 (1H,d ,J=1.5Hz,H-1″), 3.99(1H,m,H-2″), 3.84(1H,dd,J=9.5,3.5Hz,H-3″), 3.62(1H,m,H -5″), 3.59(2H,s,H-7), 3.53(2H,s,H-8), 3.44(1H,t,J=9.5Hz,H-4″), 1.20(3H,d, J=6.5Hz, H-6″). The H NMR spectrum is shown in Figure 8. ¹³ C-NMR (CD ₃ OD) δ: 158.0 (C-4'), 157.2 (C-4), 132.7 (C-1), 131.7 (C-2' and C-6'), 131.6 (C- 2 and C-6), 117.5 (C-3' and C-5'), 129.5 (C-1'), 116.2 (C-3 and C-5), 99.8 (C-1"), 73.8 (C -4"), 72.2(C-2"), 72.0(C-3"), 70.6(C-5"), 43.7(C-8), 43.4(C-7), 18.0(C-6") . The carbon NMR spectrum is shown in Figure 9. The above data indicate that compound 3 is a disulfide-containing phenolic glycoside compound.

Compound 3 was acid hydrolyzed to obtain aglycone and sugar moieties, the sugar was derivatized, and compared with the standard sugar derivative by HPLC liquid phase analysis, the sugar was identified as L-rhamnose, and the C-3- ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 3 was identified as S-[(4-O-α-L-rhamnosyl)benzyl]-S'-(4'-hydroxybenzyl)-disulfide, which was a new compound.

Compound 4 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 455.0663 [MH] ⁻ . ESI-MS is shown in Figure 10. ¹ H-NMR (CD ₃ OD) δ: 7.24 (2H, d, J=8.5 Hz, H-2 and H-6), 7.11 (2H, d, J=8.5 Hz, H-2' and H-6 '), 7.03 (1H,d, J=8.5Hz, H-3 and H-5), 6.72 (1H,d, J=8.5Hz, H-3' and H-5'), 5.40 (1H,d , J=1.5Hz, H-1″), 3.99 (2H, s, H-7), 3.98 (1H, m, H-2″), 3.94 (2H, s, H-8), 3.83 (1H, dd, J=9.5, 3.5Hz, H-3″), 3.61 (1H, m, H-5″), 3.44 (1H, t, J=9.5Hz, H-4″), 1.20 (3H, d, J=6.5Hz, H-6″). The H NMR spectrum is shown in Figure 11. ¹³ C-NMR (CD ₃ OD) δ: 158.1 (C-4), 157.3 (C-4'), 131.8 (C-1), 131.8 (C-2 and C-6), 131.7 (C-2') and C-6'), 117.6 (C-3 and C-5), 117.5 (C-3" and C-5"), 128.7 (C-1'), 99.8 (C-1"), 73.8 (C- -4"), 72.2(C-2"), 72.0(C-3"), 70.6(C-5"), 43.7(C-8), 43.4(C-7), 18.0(C-6") . The carbon NMR spectrum is shown in Figure 12. The above data indicate that compound 4 is a trisulfide-containing phenolic glycoside compound.

Compound 4 was acid hydrolyzed to obtain aglycone and sugar moiety, the sugar was derivatized, and compared with the derivative of standard sugar by HPLC liquid phase analysis, the sugar was identified as L-rhamnose, combined with C-3- of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 4 was identified as S-[(4-O-α-L-rhamnosyl)benzyl]-S"-(4'-hydroxybenzyl)-trisulfide, which was a new compound.

Example 2 Injection

Take 10 parts by weight of the compound obtained in Example 1 or its derivatives, 8.5 parts by weight of sodium chloride, and 1000 parts by weight of water for injection, mix and dissolve, stir, add 2 parts by weight of activated carbon, stir for 30min, and the solution passes through a microporous membrane (0.22 μm), filter, pack in ampoules, 5ml each, sterilize, pass the inspection, and prepare a water injection containing 50mg each. Other inspection items should meet the requirements under the "Pharmacopoeia of the People's Republic of China" (2015 edition) for injections.

Example 3 Freeze-dried powder for injection

Take 10 parts by weight of the compound obtained in Example 1 or its derivatives, dissolve it in 1000 parts by weight of water for injection containing 1% mannitol, add 5 parts by weight of activated carbon, stir for 20 min, and filter the solution through a microporous membrane (0.22 μm). , obtain a clear solution, pack in 10ml vials, 2ml each, freeze-dry, and prepare a freeze-dried powder injection containing 20mg each. Other inspection items should meet the requirements under the "Pharmacopoeia of the People's Republic of China" (2015 edition) for injections.

Example 4 Tablets

Get 20 parts by weight of the compound obtained in Example 1 or its derivative, 30 parts by weight of starch as filler, 5 parts by weight of hydroxypropyl methylcellulose as binder, and 10 parts by weight of microcrystalline cellulose as disintegrant; The lubricant is selected from 0.5 part by weight of magnesium stearate, mixed, 50% ethanol is added in an appropriate amount to granulate, dried, and pressed into tablets. Other inspection items should meet the requirements under the "Pharmacopoeia of the People's Republic of China" (2015 edition) for tablets.

Example 5 Hypoglycemic activity of thioether compounds in Moringa oleifera seeds

(1) Experimental principle

A mouse model of diabetes induced by high-fat diet (HFD) plus streptozotocin (STZ) was used, and different compounds were administered orally to observe the effects of new thioether compounds on blood sugar and body weight in diabetic mice. Effects of compounds in the treatment of diabetes.

(2) Experimental materials

Kunming mice; Moringa seed sulfide new compounds 1-4; streptozotocin; lard; blood glucose test strip and blood glucose meter; sodium citrate buffer (PH 4.4); gloves; rat box; distilled water.

(3) Experimental grouping

Experimental group of new thioether compounds; normal group (distilled water); model group (HFD+STZ).

(4) Model preparation and processing

3 male Kunming mice/cage, room temperature (22±3) ℃, relative humidity 60%±10%, daily light for 12 hours, free food and water. After 1 week of adaptive feeding, they were randomly divided into groups of 10 mice in each group: the normal group was fed with normal feed, and the high-fat diet diabetes (HFD+STZ) group was fed with high-fat feed. (HFD+STZ) mice and were given STZ solution by intraperitoneal injection at 100 mg/kg, and normal group mice were intraperitoneally injected with citric acid-sodium citrate buffer at the same dose. After STZ injection, the mice were continuously fed with the same feed, and the diet and water intake of the mice were recorded. One or two weeks after administration, blood was collected from the tail vein and detected with a blood glucose meter. The 10 mice fed with ordinary diet were the normal group, and the 60 diabetic mice that were successfully modeled were randomly divided into 6 groups. The drugs were administered according to the following groups: 20 mg/kg of the new thioether compound group in Moringa oleifera seeds, 0.3-0.4 ml/day of distilled water in the model group and normal group, administered by gavage, once a day.

(5) Specific experimental process

After 1 and 2 weeks of administration, food intake, drinking water, body weight, mental state, coat color and other conditions were observed; respectively: blood sugar level and body weight.

(6) Experimental results

①General observation

After modeling, the mice appeared polydipsia, polyphagia, polyuria, weight loss, slow response, messy and dull coat and other symptoms. Symptoms are improved to varying degrees, the response is more flexible, the hair is flat and shiny.

②Effect of new thioether compounds in Moringa seeds on blood glucose levels in HFD+STZ diabetic mice

From the results (Table 2), it can be seen that the average blood sugar of the diabetic mice induced by HFD+STZ was significantly increased, which was significantly different from that of the normal group (P<0.001). Compared with the mice in the model group, the blood glucose of the new thioether compound group in Moringa oleifera seeds was significantly decreased (P<0.05, P<0.01, P<0.001). The results showed that the new compounds of thioethers in Moringa seeds can reduce blood sugar in diabetic mice.

n＝10)The effect of thioether compounds in table 2 Moringa seeds on the blood sugar content of HFD+STZ diabetic mice (

n=10)

^### P<0.001, model group vs blank group; *P<0.05, **P<0.01, ***P<0.001, administration group vs model group

③ Effects of new thioether compounds in Moringa seeds on the body weight of HFD+STZ diabetic mice The results (Table 3) showed that the body weight of the diabetic model group was significantly higher than that of the normal group (P<0.01, P<0.001). ), the body weight of the new thioether compounds decreased significantly compared with the model group (P<0.05, P<0.01).

n＝10)The effect of thioether compounds in table 3 Moringa seeds on the body weight of diabetic mice (

n=10)

^## P<0.01, ^### P<0.001, administration group vs normal group; *P<0.05, **P<0.01, administration group vs model group.

Example 6 Antidepressant activity of thioether compounds in Moringa oleifera seeds

(1) Experimental principle

The Forced Swimming Test (FST) system is mainly used in the study of antidepressant, sedative and analgesic drugs. By placing experimental animals in a confined environment (such as water) in which the animals struggle desperately to escape but cannot escape, providing an unavoidable oppressive environment, after a period of experimentation, the animals exhibit The typical "immobility state" reflects a so-called "behavioral despair state", which records a series of parameters in the process of the animal in this environment producing a hopeless immobility state. The immobility time of mice is an indicator for judging the effect of depressive drugs. The shorter the immobility time, the stronger the antidepressant effect. Tail suspension test (TST) in mice is a classic and rapid method for evaluating the efficacy of antidepressants, stimulants and sedatives. The principle is to use the mouse to try to escape but unable to escape after hanging its tail, so as to give up the struggle and enter the unique depression and immobility state. During the experiment, the immobility time of the animal is recorded to reflect the depressive state. Shorten changes its state.

(2) Experimental materials

C57BL/6 mice; new thioether compounds in Moringa oleifera seeds; fluoxetine hydrochloride; gloves; mouse box; deionized water; plexiglass water tank; video camera;

(3) Experimental grouping

Moringa seed sulfide new compound experimental group: 10 rats in each group, 20 mg/kg; positive drug group (fluoxetine hydrochloride): 10 rats, 20 mg/kg; blank group: 10 rats, given deionized water.

(4) Specific experimental process

Forced swimming experiment: The mice were taken, fasted and kept in water for 12 hours, and moved into the laboratory 1 hour before the experiment to adapt to the environment. After 1 h of gavage, the mice were placed in glass tank water to swim, the first 2 min was used as the adaptation time, and the cumulative time of stopping swimming and floating within the next 4 min was recorded. The administration group was administered at 9-10 am every day for 6 consecutive days. On the 7th day, the mice were fasted for 1 hour before administration, and after 1 hour of gavage, the mice were placed in glass tank water to swim, and the first 2 minutes were used as the adaptation time. , the cumulative time of stopping swimming and floating within 4 minutes after recording.

Tail suspension test: 1 hour after the last administration, the part of the tail of the mouse 2 cm from the root was fixed on a self-made tail suspension bracket, so that the head of the mouse was about 5 cm away from the table top in an upside-down state, and the sides of each animal were separated by plates. open to block the sight of animals so that they do not interfere with each other. The accumulated immobility time of animals in each group within 6 minutes and 5 minutes after tail suspension was observed.

(5) Experimental results:

①Compared with the model group, the new thioether compounds in Moringa seeds can significantly shorten the cumulative immobility time of mice in forced swimming (P<0.05, P<0.01). The results are shown in Table 4.

n＝10)The effect of thioether compounds in table 4 Moringa seeds on forced swimming of mice (

n=10)

*P<0.05, **P<0.01, administration group vs blank group

②Compared with the model group, the new thioether compounds in Moringa oleifera seeds can significantly shorten the cumulative immobility time of tail-suspended mice (P<0.05). The results are shown in Table 5.

n＝10)The effect of thioether compounds in table 5 Moringa seeds on mouse tail suspension experiment (

n=10)

*P<0.05, administration group vs blank group

(6) Experimental conclusion:

The experimental results show that the thioether compounds in the Moringa seeds prepared by the present invention can shorten the immobility time of mice in the forced swimming test and the tail suspension test. The results showed that the thioether compounds prepared from Moringa oleifera seeds by a certain extraction and separation method had good antidepressant effect and could be used for the preparation and development of antidepressant preparations.

Example 7 Anti-senile dementia activity of thioether compounds in Moringa oleifera seeds

(1) Experimental principle

Aβ deposition is one of the most typical pathological features of Alzheimer's disease and plays a key role in the occurrence and development of Alzheimer's disease. According to the deposition of Aβ, the accuracy of diagnosing Alzheimer's disease can reach 80%. Studies have shown that the levels of _Aβ40 and _Aβ42 in the brain are positively correlated with the degree of cognitive impairment in patients with Alzheimer's disease. Morris water maze test is a classic method for evaluating spatial learning and memory ability, and it is an objective indicator for evaluating the replication results of animal models of dementia. In recent years, animal models of dementia have become an important means of studying senile dementia. SD rats were used to prepare Aβ _1-42 intracerebroventricular injection-induced dementia model. New thioether compounds in Moringa oleifera seeds and normal saline were administered respectively. The Morris water maze test was used to evaluate the effect of thioether compounds in Moringa oleifera seeds on Alzheimer's rats. Improvement of learning and memory ability.

(2) Experimental materials

SPF grade SD male rats; new thioether compounds in Moringa oleifera seeds; amyloid protein fragments (Aβ _1-42 ); DW-5 rat brain stereotaxic instrument; WMT-100 Morris water maze video analysis system; gloves; mice box; deionized water; plexiglass water tank; video camera; gavage needle.

(3) Experimental grouping

Control group (physiological saline), model group (condensed Aβ _1-42 ), new thioether compound group in Moringa oleifera seeds, 10 in each group.

(4) Dosage and frequency of administration

The experimental group of new thioether compounds in Moringa oleifera seeds was administered 20 mg/kg by gavage, once a day for 7 consecutive days.

(5) Specific experimental process

①Model preparation: SD rats were anesthetized with 10% urethane, and the head was fixed in a flat head position. The stereotaxic instrument was positioned at 3.4 mm posterior to the bregma, 2.0 mm lateral to the midline of the brain, and 2.7 mm below the skull surface. 5 μL of condensed Aβ _1-42 were injected slowly and uniformly at mm respectively. The Aβ model control group was injected with an equal volume of isotonic saline. Morris water maze behavior test was performed 1 week after modeling.

②Morris water maze behavioral test: choose a quiet, dark, and constant temperature environment for the test. Different graphics can be appropriately pasted on the walls of the laboratory to help animals determine their orientation. Before the experiment, the bucket was filled with water to a predetermined height (about 40cm), and then an appropriate amount of white pigment was added to make the water an opaque milky white liquid. The heater heated the water temperature to 25°C. The platform was placed in the center of the fourth quadrant, about 1.5 cm below the water surface, and the platform position remained unchanged throughout the experiment. Positioning Voyage Test: Positioning Voyage Test lasts for four days. The platform was placed in the center of the second quadrant, and the rats entered the water from the four quadrants facing the pool wall in turn, and the time it took for the rats to find the platform within 120 s was recorded, that is, the escape latency (EL). If the rat does not find the platform within 120 s, the escape latency is 120 s, and the rat should be guided to board the platform at this time. All rats on the platform should stay on the platform for 15 s, so that they recognize the underwater platform as an escape point and memorize the spatial location of the platform. The average escape latency of each group of rats in the four quadrants per day was calculated, that is, the mean escape latency. The average escape latency (AEL) reflects the learning and memory ability of the rats. The shorter the AEL, the faster the learning and cognition of the spatial location of the underwater platform. Space search test: The space search test is carried out 24 hours after the end of the positioning navigation test. The specific method is to remove the platform, let the rat search for the platform by memory without the platform, record the number of times the rat crosses the platform, the swimming distance in each quadrant and the total distance within 120s. Throughout the test, the position of the reference objects around the pool, including the experimenter, should be fixed.

(6) Experimental results

As shown in Table 6, in the positioning navigation test, compared with the model group, the average escape latency of thioether compounds in Moringa oleifera seeds was significantly shortened (P<0.05). In the spatial search test, compared with the model group, the cross-platform times and distances of the thioether compound group in Moringa seeds were significantly increased (P<0.05, P<0.01) (Table 7). The above results show that the thioether compounds in Moringa oleifera seeds can improve the memory impairment of senile dementia rats.

n＝10)The effect of thioether compounds in table 6 Moringa seeds on the escape latency of Aβ _1-42 -induced senile dementia rats (

n=10)

^# P<0.05, ^## P<0.01, model group vs control group; *P<0.05, administration group vs model group

n＝10)The effect of thioether compounds in table 7 Moringa seeds on Aβ _1-42 -induced senile dementia rats spatial search experiment (

n=10)

^## P<0.01, model group vs control group; *P<0.05, **P<0.01, administration group vs model group

Claims (2)

1. thioether compounds in Moringa oleifera seeds, is characterized in that being selected from disulfide or trisulfide compounds, and concrete structure is as follows:

2. the application of the thioether compound in the Moringa seed described in claim 1 in the preparation and treatment of diabetes, antidepressant and anti-senile dementia medicine.

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