CN109160928B - New phenolic glycosides from Moringa seeds and their applications - Google Patents

Abstract

The present invention relates to 15 new phenolic glycoside compounds extracted and separated from Moringa oleifera seeds, the general structural formula of such compounds is shown in (I), and the present invention also relates to the preparation of such compounds in the preparation and treatment of diabetes, antidepressant and anti-diabetic drugs. Application of Alzheimer's drugs and health care products. Experiments show that the compounds of the present invention have significant hypoglycemic, antidepressant and anti-senile dementia effects. The compound of the invention has the characteristics of clear active mechanism, low toxicity, safety and the like, and has broad application prospects.

Description

New phenolic glycosides from Moringa seeds and their applications

technical field

The invention relates to a new phenolic glycoside compound separated from Moringa seeds and its application in the preparation of medicines for treating diabetes, antidepression and anti-senile dementia and health care products.

Background technique

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 planted 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.

The present invention discovers a series of phenolic compounds from Moringa oleifera seeds, including 15 new compounds, which are found for the first time through research to have the functions of treating diabetes, anti-depression and anti-senile dementia.

SUMMARY OF THE INVENTION

The invention relates to a new phenolic glycoside compound separated from Moringa oleifera seeds and its application in the preparation of medicines for treating diabetes, anti-depression and anti-senile dementia and health care products.

The general chemical formula of the novel phenolic glycosides and derivatives thereof of the present invention is as follows:

Wherein R ₁ is hydrogen or sugar group; R ₂ is ester group or thioester, urea or thiourea, oxygen ether, etc. The ester group is preferably methyl formate, ethyl formate, butyl formate, acetate, Butyrate group; thioester preferably sulfur-substituted methyl formate, sulfur-substituted methyl carbamate, sulfur-substituted ethyl formate, sulfur-substituted ethyl carbamate, sulfur-substituted propyl formate urea, sulfur-substituted methylpropyl carbamate, sulfur-substituted butyl carbamate, sulfur-substituted butyl carbamate, sulfur-substituted aryl formate, sulfur-substituted aryl carbamate; urea Preferred are urea, thiourea, semicarbazide, thiosemicarbazide, aryl urea, aryl thiourea, etc.; oxygen ethers are preferably methyl ether, ethyl ether, propyl ether, butyl ether and other alkanes and aryl ethers, etc.; sugar The group is preferably a six-carbon (glucose, mannose, rhamnose) pyranose, a six-carbon furanose, and the acylated sugar is preferably a six-carbon (glucose, mannose, rhamnose) pyranose, a six-carbon furanose, or an acyl group 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 new phenolic glycosides

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:

Figure 1 shows the ESI-MS image of compound 1

Figure 2 is the H NMR spectrum of compound 1

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

Figure 4 is the ESI-MS image 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

Figure 7 is the ESI-MS image 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 image of compound 4

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

Figure 12 is the carbon nuclear magnetic resonance spectrum of compound 4

Figure 13 is the ESI-MS image of compound 5

Figure 14 is the hydrogen nuclear magnetic resonance spectrum of compound 5

Figure 15 is the carbon nuclear magnetic resonance spectrum of compound 5

Figure 16 is the ESI-MS image of compound 6

Figure 17 is the hydrogen nuclear magnetic resonance spectrum of compound 6

Figure 18 is the carbon nuclear magnetic resonance spectrum of compound 6

Figure 19 is the ESI-MS image of compound 7

Figure 20 is the hydrogen nuclear magnetic resonance spectrum of compound 7

Figure 21 is the carbon nuclear magnetic resonance spectrum of compound 7

Figure 22 is the ESI-MS image of compound 8

Figure 23 is the hydrogen nuclear magnetic resonance spectrum of compound 8

Figure 24 is the carbon nuclear magnetic resonance spectrum of compound 8

Figure 25 is the ESI-MS image of compound 9

Figure 26 is the hydrogen nuclear magnetic resonance spectrum of compound 9

Figure 27 is the carbon nuclear magnetic resonance spectrum of compound 9

Figure 28 is the ESI-MS image of compound 10

Figure 29 is the hydrogen nuclear magnetic resonance spectrum of compound 10

Figure 30 is the carbon nuclear magnetic resonance spectrum of compound 10

Figure 31 is the ESI-MS image of compound 11

Figure 32 is the hydrogen nuclear magnetic resonance spectrum of compound 11

Figure 33 is the carbon nuclear magnetic resonance spectrum of compound 11

Figure 34 is the ESI-MS image of compound 12

Figure 35 is the hydrogen nuclear magnetic resonance spectrum of compound 12

Figure 36 is the carbon nuclear magnetic resonance spectrum of compound 12

Figure 37 is the ESI-MS image of compound 13

Figure 38 is the hydrogen nuclear magnetic resonance spectrum of compound 13

Figure 39 is the carbon nuclear magnetic resonance spectrum of compound 13

Figure 40 is the ESI-MS image of compound 14

Figure 41 is the hydrogen nuclear magnetic resonance spectrum of compound 14

Figure 42 is the carbon nuclear magnetic resonance spectrum of compound 14

Figure 43 is the ESI-MS image of compound 15

Figure 44 is the hydrogen nuclear magnetic resonance spectrum of compound 15

Figure 45 is the carbon nuclear magnetic resonance spectrum of compound 15

Fourth, the specific implementation

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 10kg of Moringa seeds and pulverize, add 10 times the weight of 30% ethanol for diafiltration and extraction twice, each time for 24h, 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 were eluted with 80 L of 85% ethanol, the 85% ethanol eluate was recovered, concentrated, and dried under reduced pressure to obtain the total extract. 350 g of the total extract was subjected to silica gel column chromatography, eluted with a gradient of chloroform-methanol system (the volume ratio of chloroform and methanol from 100:0 to 0:1), and the solvent was recovered for each gradient to obtain fraction Fr.1 -Fr.8. Fr.3 (18g) was subjected to ODS (reverse octadecyl bonded silica gel) column chromatography, methanol-water system gradient elution (methanol-water volume ratio from 2:8 to 1:0) to obtain 10 fractions, Fr.3a-Fr.3j, wherein Fr.3c (0.7 g) was subjected to preparative HPLC with 30% methanol-water as mobile phase to obtain compound 1 (350 mg). Fr.3e (1.1 g) was subjected to preparative HPLC with 43% methanol-water as mobile phase to obtain compound 3 (215 mg). Fr.3f (1.3 g) was subjected to preparative HPLC with 45% methanol-water as mobile phase to obtain compound 4 (260 mg). Fr.3 g (0.65 g) was subjected to preparative HPLC with 28% methanol-water as mobile phase to obtain compound 13 (420 mg). Fr.3i (0.86 g) was subjected to preparative HPC with 50% methanol-water as mobile phase to obtain compound 8 (308 mg). Fr.5 (20g) was subjected to silica gel column chromatography, eluted with a gradient of cyclohexane-ethyl acetate system (volume ratio of cyclohexane to ethyl acetate from 100:0 to 1:1), and the eluents were combined to obtain 6 Fractions, Fr.5a-Fr.5f, of which Fr.5a (1.2 g) were subjected to preparative HPLC with 60% methanol-water as mobile phase to obtain compound 6 (365 mg). Fr.5c (0.8 g) was subjected to preparative HPLC with 48% methanol-water as mobile phase to obtain compound 11 (342 mg). Fr.5d (1.1 g) was subjected to preparative HPLC with 55% methanol-water as mobile phase to obtain compound 9 (517 mg). Fr.5e (2.1 g) was subjected to preparative HPLC with 38% methanol-water as mobile phase to obtain compounds 12 (338 mg) and 14 (305 mg). Fr.6 (25g) was subjected to ODS column chromatography, methanol-water system gradient elution (methanol-water volume ratio from 15:85 to 1:0) to obtain 7 fractions, Fr.6a-Fr.6g, in which Fr. Compound 15 (345 mg) was prepared by preparative HPLC of 6a (0.9 g) with 30% methanol-water as mobile phase. Fr.6c (1.2 g) was subjected to preparative HPLC with 43% methanol-water as mobile phase to obtain compounds 2 (503 mg) and 7 (384 mg). Fr.6e (0.6 g) was subjected to preparative HPLC with 63% methanol-water as mobile phase to obtain compound 5 (407 mg). Fr.6c (1.1 g) was subjected to preparative HPLC with 38% methanol-water as mobile phase to obtain compound 10 (461 mg).

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

Spectral data are as follows: ESI-MS m/z: 297.0972 [MH] ⁻ . ESI-MS is shown in Figure 1. ¹ H-NMR (CD ₃ OD) δ: 7.20 (2H, d, J=8.5 Hz, H-2 and H-6), 7.01 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.41(1H,d,J=1.5Hz,H-1'), 4.00(1H,m,H-2'), 3.86(1H,dd,J=9.5,3.5Hz,H-3'), 3.64 (1H,m,H-5'), 3.54(2H,s,H-7), 3.46(1H,t,J=9.5Hz,H-4'), 1.23(3H,d,J=6.5Hz, H-6'). The H NMR spectrum is shown in Figure 2. ¹³ C-NMR (CD ₃ OD) δ: 175.8 (C-9), 156.8 (C-4), 131.4 (C-1), 129.7 (C-2 and C-6), 117.5 (C-3 and C -5), 99.8(C-1'), 73.8(C-4'), 72.2(C-3'), 72.0(C-2'), 70.6(C-5'), 41.0(C-7) , 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 3.

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 was identified as O-benzyl formate-4-O-α-L-rhamnoside, which was a new compound.

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

Spectral data are as follows: ESI-MS m/z: 371.1357 [M+HCOO] ⁻ . ESI-MS is shown in Figure 4. ¹ H-NMR (CD ₃ OD) δ: 7.14 (2H, d, J=8.5 Hz, H-2 and H-6), 6.95 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.34 (1H, d, J=1.5Hz, H-1'), 4.08 (1H, q, J=7.1Hz, H-9), 3.93 (1H, m, H-2'), 3.77 (1H, dd, J = 9.5, 3.5 Hz, H-3'), 3.58 (1H, m, H-5'), 3.51 (2H, s, H-7), 3.39 (1H, t, J = 9.5 Hz, H -4'), 1.18 (3H, d, J=6.5 Hz, H-6'), 1.16 (1H, t, J=7.1 Hz, H-10). The H NMR spectrum is shown in Figure 5. ¹³ C-NMR (CD ₃ OD) δ: 173.8 (C-8), 156.9 (C-4), 131.4 (C-1), 129.4 (C-2 and C-6), 117.6 (C-3 and C -5), 99.9(C-1'), 73.9(C-4'), 72.2(C-2'), 72.0(C-3'), 70.6(C-5'), 61.6(C-9) , 41.2 (C-7), 18.0 (C-6'), 14.5 (C-10). The carbon NMR spectrum is shown in Figure 6.

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 is identified as O-ethyl-ethyl phenylacetate-4-O-α-L-rhamnoside, which is a new compound.

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

Spectral data are as follows: ESI-MS m/z: 399.1635 [M+HCOO] ⁻ . ESI-MS is shown in Figure 7. ¹ H-NMR (CD ₃ OD) δ: 7.20 (2H, d, J=8.7 Hz, H-2 and H-6), 6.95 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.40 (1H, d, J=1.8Hz, H-1'), 4.08 (1H, t, J=6.6Hz, H-9), 3.99 (1H, m, H-2'), 3.83 (1H, dd, J = 9.5, 3.5 Hz, H-3'), 3.63 (1H, m, H-5'), 3.58 (2H, s, H-7), 3.45 (1H, t, J = 9.5 Hz, H -4′), 1.59 (2H, m, H-10), 1.33 (2H, m, H-11), 1.18 (3H, d, J=6.2Hz, H-6′), 0.92 (1H, t, J=7.4Hz, H-12). The H NMR spectrum is shown in Figure 8. ¹³ C-NMR (CD ₃ OD) δ: 173.8 (C-8), 156.9 (C-4), 131.4 (C-1), 129.4 (C-2 and C-6), 117.5 (C-3 and C -5), 99.8(C-1'), 73.8(C-4'), 72.2(C-2'), 72.1(C-3'), 70.6(C-5'), 65.9(C-9) , 41.2 (C-7), 31.8 (C-10), 20.1 (C-11), 18.0 (C-6'), 14.5 (C-12). The carbon NMR spectrum is shown in Figure 9.

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 O-butyl-phenylacetate-4-O-α-L-rhamnoside, 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: 403.1043 [M+HCOO] ⁻ . ESI-MS is shown in Figure 10. ¹ H-NMR (CD ₃ OD) δ: 7.27 (2H, d, J=8.7 Hz, H-2 and H-6), 7.20 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.40 (1H,d,J=1.6Hz,H-1', 4.26(1H,q,J=7.1Hz,H-9), 4.06(2H,s,H-7), 3.98(1H,m, H-2'), 3.84(1H,dd,J=9.5,3.4Hz,H-3'), 3.62(1H,m,H-5'), 3.45(1H,t,J=9.5Hz,H- 4′), 1.27 (1H, t, J=7.1Hz, H-10), 1.22 (3H, d, J=6.2Hz, H-6′). H NMR spectrum is shown in Figure 11. ¹³ C- NMR (CD ₃ OD) δ: 172.0 (C-8), 157.1 (C-4), 132.7 (C-1), 131.1 (C-2 and C-6), 117.5 (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'), 64.6(C-9), 35.4 ( C-7), 18.0 (C-6'), 14.6 (C-10). The carbon nuclear magnetic resonance spectrum is shown in Figure 12.

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 O-ethyl-benzylthiocarboxylate ethyl ester-4-O-α-L-rhamnoside, which was a new compound.

Compound 5 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 431.1355 [M+HCOO] ⁻ . ESI-MS is shown in Figure 13. ¹ H-NMR (CD ₃ OD) δ: 7.27 (2H, d, J=8.6 Hz, H-2 and H-6), 6.99 (1H, d, J=8.6 Hz, H-3 and H-5) , 5.40 (1H,d,J=1.5Hz,H-1'), 4.23(2H,d,J=6.6Hz,H-9), 4.06(1H,m,H-7), 3.98(1H,m ,H-2'), 3.84(1H,dd,J=9.5,3.4Hz,H-3'),3.63(1H,m,H-5'),3.45(1H,t,J=9.5Hz,H -4'), 1.63(2H,m,H-10), 1.38(2H,m,H-11), 1.22(3H,d,J=6.2Hz,H-6'), 0.95(3H,t, J=7.4Hz, H-12). The H NMR spectrum is shown in Figure 14. ¹³ C-NMR (CD ₃ OD) δ: 172.2 (C-9), 157.1 (C-4), 132.7 (C-1), 131.1 (C-2 and C-6), 117.6 (C-3 and C -5), 98.9(C-1'), 73.8(C-4'), 72.2(C-2'), 72.0(C-3'), 70.6(C-5'), 68.4(C-9) , 35.4 (C-7), 31.9 (C-10), 20.0 (C-11), 18.0 (C-6'), 14.0 (C-12). The carbon NMR spectrum is shown in Figure 15.

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

Based on the above information, compound 5 was identified as butyl O-butyl-benzylthiocarboxylate-4-O-α-L-rhamnoside, which is a new compound.

Compound 6 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 402.1224 [M+HCOO] ⁻ . ESI-MS is shown in Figure 16. ¹ H-NMR (CD ₃ OD) δ: 7.14 (2H, d, J=8.6 Hz, H-2 and H-6), 6.94 (1H, d, J=8.6 Hz, H-3 and H-5) , 5.33 (1H, d, J=1.5Hz, H-1'), 4.25 (2H, s, H-7), 3.91 (1H, m, H-2'), 3.75 (1H, dd, J=9.5 , 3.4Hz, H-3'), 3.54(1H,m,H-5'), 3.37(1H,t,J=9.5Hz,H-4'), 2.80(2H,q,J=7.3Hz, H-9), 1.18 (3H, t, J=7.3 Hz, H-10), 1.14 (3H, d, J=6.2 Hz, H-6'). The H NMR spectrum is shown in Figure 16. ¹³ C-NMR (CD ₃ OD) δ 157.0 (C-4), 133.7 (C-1), 129.8 (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'), 45.0(C-7), 24.7(C-9) , 18.0 (C-6'), 16.3 (C-10). The carbon NMR spectrum is shown in Figure 18.

Compound 6 was acid hydrolyzed to obtain aglycon and sugar moieties, 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, and the C-3— ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 6 was identified as S-ethyl-benzylcarbamate-4-O-α-L-rhamnoside, which is a new compound.

Compound 7 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 563.2242 [M+HCOO] ⁻ . ESI-MS is shown in Figure 1. ¹ H-NMR (DMSO-d ₆ ) δ: 7.18 (2H, d, J=8.5Hz, H-2 and H-6), 6.98 (1H, d, J=8.5Hz, H-3 and H-5 ), 5.33(1H,d,J=1.5Hz,H-1'), 4.16(2H,d,J=5.9Hz,H-7), 3.82(1H,m,H-2'), 3.64(1H ,dd, J=9.3,3.5Hz,H-3'), 3.45(1H,m,H-5'), 3.28(1H,t,J=9.3Hz,H-4'), 1.10(3H,d , J=6.2 Hz, H-6′). The H NMR spectrum is shown in Figure 2. ¹³ C-NMR (DMSO-d ₆ ) δ: 158.1 (C-8), 154.9 (C-4), 134.3 (C-1), 128.3 (C-2 and C-6), 116.4 (C-3 and C-5), 98.5(C-1'), 71.9(C-4'), 70.5(C-2'), 70.3(C-3'), 69.5(C-5'), 42.6(C-7 ), 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 3. According to the above data, compound 7 is a highly symmetrical phenolic glycoside compound containing urea group.

Compound 7 was acid hydrolyzed to obtain aglycone and sugar moieties, and the sugar was subjected to derivatization reaction. The HPLC liquid phase analysis was compared with the derivatives of standard sugar, and the sugar was identified as L-rhamnose, combined with the C-3-rhamnose of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 7 was identified as N,N'-bis[(4-O-α-L-rhamnosyl)benzyl]-urea, which was a new compound.

Compound 8 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 433.1435 [MH] ⁻ . ESI-MS is shown in Figure 4. ¹ H-NMR (CD ₃ OD) δ: 7.21 (2H, d, J=8.5 Hz, H-2' and H-6'), 7.11 (2H, d, J=8.5 Hz, H-2 and H- 6), 7.01 (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″), 3.98 (1H,m,H-2″), 3.84 (1H,dd,J=9.5,3.5Hz,H-3″), 3.62 (1H,m,H -5"), 3.45 (1H, t, J=9.5Hz, H-4"), 1.20 (3H, d, J=6.5Hz, H-6"). The H NMR spectrum is shown in Figure 5. ¹³ C-NMR (CD ₃ OD) δ: 157.8 (C-4'), 157.0 (C-4), 130.0 (C-2' and C-6'), 129.8 (C-2 and C-6), 117.5 (C-3' and C-5'), 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"), 18.0 (C-6"). The carbon NMR spectrum is shown in Figure 6. According to the above data, compound 8 is a phenolic glycoside compound containing a urea group.

Compound 8 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, combined with the C-3-rhamnose of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 8 was identified as N-[(4-O-α-L-rhamnosyl)benzyl]-N'-(4'-hydroxybenzyl)-thiourea, which was a new compound.

Compound 9 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 342.1123 [MH] ⁻ . ESI-MS is shown in Figure 7. ¹ H-NMR (CD ₃ OD) δ: 7.22 (2H, d, J=8.7 Hz, H-2 and H-6), 6.01 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.38(1H,d,J=1.8Hz,H-1'), 4.62(2H,s,H-7), 3.94(1H,m,H-2'), 3.78(1H,dd,J=9.5 , 3.5Hz, H-3′), 3.55 (1H, m, H-5′), 3.40 (1H, t, J=9.5Hz, H-4′), 1.16 (3H, d, J=6.2Hz, H-6'). The H NMR spectrum is shown in Figure 8. ¹³ C-NMR (CD ₃ OD) δ: 157.7 (C-4), 129.6 (C-2 and C-6), 117.8 (C-3 and C-5), 99.7 (C-1'), 73.7 ( C-4'), 72.1 (C-2'), 71.9 (C-3'), 70.7 (C-5'), 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 9. According to the above data, compound 9 is a phenolic glycoside compound containing a thiourea group.

Compound 9 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 C-3-rhamnose of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 9 was identified as N-[(4-O-α-L-rhamnosyl)benzyl]-N'-amino-thiourea, which was a new compound.

Compound 10 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 357.1318 [M+HCOO] ⁻ . ESI-MS is shown in Figure 10. ¹ H-NMR (CD ₃ OD) δ: 7.22 (2H, d, J=8.5 Hz, H-2 and H-6), 7.01 (1H, d, J=8.5 Hz, H-3 and H-5) , 5.40 (1H, d, J=1.5Hz, H-1'), 4.24 (2H, s, H-7), 3.99 (1H, m, H-2'), 3.84 (1H, dd, J=9.5 , 3.5Hz, H-3′), 3.63(1H,m,H-5′), 3.45(1H,t,J=9.5Hz,H-4′), 1.21(3H,d,J=6.5Hz, H-6'). The H NMR spectrum is shown in Figure 11. ¹³ C-NMR (CD ₃ OD) δ: 162.1 (C-8), 157.0 (C-4), 134.8 (C-1), 129.6 (C-2 and C-6), 117.6 (C-3 and C -5), 99.9(C-1'), 73.9(C-4'), 72.2(C-3'), 72.1(C-2'), 70.6(C-5'), 44.2(C-7) , 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 12. According to the above data, compound 10 is a phenolic glycoside compound containing a urea group.

Compound 10 was acid hydrolyzed to obtain aglycone and sugar moiety, the sugar was derivatized, and compared with the standard sugar derivative by HPLC liquid phase analysis, the sugar was identified as L-rhamnose, combined with the C-3-rhamnose of rhamnose. ^The13C chemical shift value of C-5 identified this glycosyl as α-L-rhamnosyl.

Based on the above information, compound 10 was identified as N-[(4-O-α-L-rhamnosyl)benzyl]-urea, which was a new compound.

Compound 11 is a yellow amorphous powder with the chemical structural formula of

Spectral data are as follows: ESI-MS m/z: 414.1771 [M+HCOO] ⁻ . ESI-MS is shown in Figure 7. ¹ H-NMR (CD ₃ OD) δ: 7.15 (2H, d, J=8.6 Hz, H-2 and H-6), 6.96 (1H, d, J=8.6 Hz, H-3 and H-5) , 5.32 (1H, d, J=1.5Hz, H-1'), 4.09 (2H, d, J=6.1Hz, H-7), 3.80 (1H, m, H-2'), 3.94 (2H, t, J=6.6 Hz, H-9), 3.61 (1H, dd, J=9.3, 3.5 Hz, H-3'), 3.43 (1H, m, H-5'), 3.26 (1H, t, J =9.3Hz, H-4'), 1.51(2H,m,H-10), 1.31(2H,m,H-11), 1.06(3H,d,J=6.1Hz,H-6'), 0.87 (3H, t, J = 7.4 Hz, H-12). The H NMR spectrum is shown in Figure 8. ¹³ C-NMR (CD ₃ OD) δ: 156.6 (C-8), 155.0 (C-4), 133.3 (C-1), 128.4 (C-2 and C-6), 116.3 (C-3 and C -5), 98.4(C-1'), 71.8(C-4'), 70.5(C-2'), 70.3(C-3'), 69.6(C-5'), 63.6(C-9) , 43.2 (C-7), 30.8 (C-10), 18.7 (C-11), 18.0 (C-6'), 13.7 (C-12). The carbon NMR spectrum is shown in Figure 9.

Compound 11 was acid hydrolyzed to obtain the aglycone and sugar moiety, 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 11 was identified as O-butyl-benzylcarbamate-4-O-α-L-rhamnoside, which was a new compound.

Compound 12 is a yellow amorphous powder with the chemical structural formula of

(CD ₃ OD) δ: 7.15 (2H, d, J=8.6 Hz, H-2 and H-6), 6.96 (1H, d, J=8.6 Hz, H-3 and H-5), 5.32 (1H ,d,J=1.5Hz,H-1'), 4.09(2H,d,J=6.1Hz,H-7), 3.80(1H,m,H-2'), 3.94(2H,t,J= 6.6Hz, H-9), 3.61 (1H, dd, J=9.3, 3.5Hz, H-3'), 3.43 (1H, m, H-5'), 3.26 (1H, t, J=9.3Hz, H-4'), 1.51(2H,m,H-10), 1.31(2H,m,H-11), 1.06(3H,d,J=6.1Hz,H-6'), 0.87(3H,t , J=7.4Hz, H-12). The H NMR spectrum is shown in Figure 8. ¹³ C-NMR (CD ₃ OD) δ: 156.6 (C-8), 155.0 (C-4), 133.3 (C-1), 128.4 (C-2 and C-6), 116.3 (C-3 and C -5), 98.4(C-1'), 71.8(C-4'), 70.5(C-2'), 70.3(C-3'), 69.6(C-5'), 63.6(C-9) , 43.2(C-7), 30.8(C-10), 18.7(C-11), 18.0(C-6'), 13.7(C-12). The carbon NMR spectrum is shown in Figure 9.

Compound 12 was acid hydrolyzed to obtain the aglycone and sugar moiety, 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 12 was identified as O-butyl-benzylcarbamate-4-O-α-L-rhamnoside, which was a new compound.

Compound 13 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 283.0309 [MH] ⁻ . ESI-MS is shown in Figure 1. ¹ H-NMR (CD ₃ OD) δ: 7.89 (2H, d, J=8.8 Hz, H-2 and H-6), 7.05 (1H, d, J=8.8 Hz, H-3 and H-5) , 5.46(1H,d,J=1.5Hz,H-1'), 3.93(1H,m,H-2'), 3.76(1H,dd,J=9.5,3.5Hz,H-3'), 3.50 (1H, m, H-5'), 3.39 (1H, t, J=9.5 Hz, H-4'), 1.15 (3H, d, J=6.2 Hz, H-6'). The H NMR spectrum is shown in Figure 2. ¹³ C-NMR (CD ₃ OD) δ: 169.6 (C-7), 161.5 (C-4), 125.4 (C-1), 132.8 (C-2 and C-6), 116.9 (C-3 and C -5), 99.5(C-1'), 73.6(C-4'), 72.1(C-3'), 71.8(C-2'), 70.9(C-5'), 18.0(C-6') ). The carbon NMR spectrum is shown in Figure 3.

Compound 13 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 13 was identified as 4-O-α-L-rhamnosylbenzoic acid, which was a new compound.

Compound 14 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 445.1357 [MH] ⁻ . ESI-MS is shown in Figure 4. ¹ H-NMR (CD ₃ OD) δ: 7.95 (2H, d, J=8.8 Hz, H-2 and H-6), 6.12 (1H, d, J=8.8 Hz, H-3 and H-5) , 5.33(1H,d,J=1.5Hz,H-1′), 4.58(1H,d,J=7.7Hz,H-1″), 4.29(1H,m,H-2′), 3.92(1H ,m,H-3′), 3.82(1H,m,H-6″), 3.71(1H,dd,J=11.9,4.6Hz,H-6″), 3.61(1H,m,H-4′ ), 3.60(1H,m,H-5′), 3.37(1H,m,H-5″), 3.35(1H,m,H-4″), 3.31(1H,m,H-2″), 3.30 (1H, m, H-3"), 1.20 (3H, d, J=5.8Hz, H-6'). The hydrogen nuclear magnetic resonance spectrum is shown in Figure 5. ¹³ C-NMR (CD ₃ OD)δ: 169.5 (C-7), 161.4 (C-4), 125.5 (C-1), 132.7 (C-2 and C-6), 116.9 (C-3 and C-5), 105.9 (C-1") , 99.2(C-1'), 82.6(C-3'), 77.7(C-5"), 77.6(C-2"), 75.4(C-3"), 72.4(C-4'), 71.2 (C-2'), 70.9 (C-4"), 70.5 (C-5'), 62.1 (C-6"), 18.1 (C-6'). The carbon nuclear magnetic resonance spectrum is shown in FIG. 6 .

Compound 14 was acid hydrolyzed to obtain the aglycone and sugar moiety, the sugar was derivatized, and compared with the standard sugar derivative by HPLC liquid phase analysis, the sugar was identified as D-grape and L-rhamnose, according to the glucose terminal proton The coupling constant (J=7.7Hz) can be determined as β-D-glucose, and the glycosyl group is identified as α-L-rhamnosyl based on the ¹³ C chemical shift values of C-3-C-5 of rhamnose.

Based on the above information, compound 14 was identified as 4-O-β-D-glucose-(1→3)-α-L-rhamnosylbenzoic acid, which was a new compound.

Compound 15 is a yellow amorphous powder with the chemical structural formula:

Spectral data are as follows: ESI-MS m/z: 298.1293 [M] ⁺ . ESI-MS is shown in Figure 10. ¹ H-NMR (D ₂ O) δ: 7.45 (2H, d, J=8.7 Hz, H-2 and H-6), 7.20 (1H, d, J=8.7 Hz, H-3 and H-5) , 5.61(1H,d,J=1.5Hz,H-1'), 4.19(1H,m,H-2'), 4.16(2H,s,H-7), 4.02(1H,dd,J=9.5 ,3.5Hz,H-3'), 3.81(1H,m,H-5'), 3.54(1H,t,J=9.5Hz,H-4'), 1.92(3H,s,H-8), 1.24 (3H, d, J=6.2 Hz, H-6'). The H NMR spectrum is shown in Figure 11. ¹³ C-NMR (D ₂ O) δ: 155.8 (C-4), 130.6 (C-2 and C-6), 127.2 (C-1), 117.5 (C-3 and C-5), 98.0 (C -1'), 71.9(C-4'), 70.1(C-3'), 69.9(C-2'), 69.5(C-5'), 42.5(C-7), 23.4(C-8) , 18.0 (C-6'). The carbon NMR spectrum is shown in Figure 12.

Compound 15 was acid hydrolyzed to obtain aglycone and sugar moiety, and the sugar was subjected to derivatization reaction. The derivation of standard sugar was compared by HPLC liquid phase analysis, and 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 15 was identified as N-oxidative-methyl-phenethylamine-4-O-α-L-rhamnoside, 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), A clear solution was obtained, which was divided into 10ml vials, 2ml each, and freeze-dried to 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 phenolic glycosides in Moringa 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 phenolic glycosides on blood sugar and body weight of diabetic mice. Effects of new phenolic glycosides compounds from Moringa oleifera in the treatment of diabetes.

(2) Experimental materials

Kunming mice; Moringa seed phenolic glycosides 1-15; streptozotocin; lard; blood glucose test strip and blood glucose meter; sodium citrate buffer (PH 4.4); gloves; rat box; distilled water.

(3) Experimental grouping

Phenolic glycoside new compound experimental group; 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. 10 mice fed with normal diet were used as normal group, and 50 diabetic mice with successful modeling were randomly divided into 5 groups. The drugs were administered according to the following groups: 30 mg/kg of the new Moringa phenol glycosides group, 0.3-0.4 ml/day of distilled water in the model group and the normal group, administered by gavage, once a day.

(5) Specific experimental process

After 1 and 2 weeks of administration, the 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, unresponsiveness, messy and dull coat and other symptoms. After administration of the new phenolic glycoside compounds in Moringa oleifera Symptoms are improved to varying degrees, the response is more flexible, the hair is flat and shiny.

②Effects of new phenolic glycosides from 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 model group, the blood sugar of the moringa seed phenolic glycoside new compound group was significantly lower (P<0.05, P<0.01). The results showed that the new compounds of Moringa phenolic glycosides can reduce blood sugar in diabetic mice.

Table 2 Effects of Moringa oleifera phenolic glycosides on blood glucose levels in HFD+STZ diabetic mice

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

③The effect of new Moringa phenolic glycosides on the body weight of HFD+STZ diabetic mice It can be seen from the results (Table 3) that the body weight of the mice in the diabetes model group was significantly higher than that of the mice in the drug treatment group (P<0.01, P<0.01, P<0.01). 0.001), the body weight of the new Moringa phenolic glycosides decreased significantly compared with the model group (P<0.05).

Table 3 Effects of Moringa oleifera phenolic glycosides on body weight of diabetic mice

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

Example 6 Antidepressant activity of phenolic glycosides in Moringa 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 phenolic glycosides in Moringa seeds; fluoxetine hydrochloride; gloves; mouse box; deionized water; plexiglass water tank; video camera;

(3) Experimental grouping

Experimental group of phenolic glycosides in Moringa oleifera seeds: 10 animals in each group, 20 mg/kg; positive drug group (fluoxetine hydrochloride): 10 animals, 20 mg/kg; blank group: 10 animals, 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 compounds of phenolic glycosides in Moringa seeds can significantly shorten the cumulative immobility time of mice in forced swimming (P<0.01). The results are shown in Table 4.

Table 4 Effects of phenolic glycosides in Moringa seeds on forced swimming in mice

**P<0.01, administration group vs blank group

②Compared with the model group, the new compounds of phenolic glycosides 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.

Table 5 Effects of phenolic glycosides in Moringa seeds on mouse tail suspension experiment

*P<0.05, administration group vs blank group.

(6) Experimental conclusion:

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

Example 7 Anti-senile dementia activity of phenolic glycosides of Moringa oleifera

(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 a model of dementia induced by Aβ _1-42 intracerebroventricular injection. New phenolic glycosides in Moringa seeds and normal saline were administered respectively. Morris water maze test was used to evaluate the effect of phenolic glycosides in Moringa seeds on Alzheimer's rats Improvement of learning and memory ability.

(2) Experimental materials

SPF grade SD male rats; new phenolic glycosides in Moringa seeds; amyloid 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 ), experimental group of new phenolic glycosides in Moringa oleifera seeds. 10 per group.

(4) Dosage and frequency of administration

The experimental group of new phenolic glycosides in Moringa 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 clean water to a predetermined height (about 40 cm), 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 was to remove the platform, let the rat search for the platform by memory without the platform, and record the number of times the rat crossed the platform within 120s, the swimming distance in each quadrant and the total distance. 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 the new phenolic glycosides of Moringa oleifera was significantly shortened (P<0.05, P<0.01). In the spatial search test, compared with the model group, the cross-platform times and distances in the high-dose group of phenolic glycosides from Moringa oleifera were significantly increased (P<0.05, P<0.01) (Table 7). The above results show that: Moringa seed phenolic glycosides have the effect of improving memory impairment in senile dementia rats.

Table 6 Effects of Moringa oleifera phenolic glycosides on escape latency of Aβ _1-42 -induced senile dementia rats

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

Table 7 Effects of Moringa oleifera phenolic glycosides on Aβ _1-42 -induced senile dementia rats spatial search experiment

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

It should be pointed out that the above-mentioned embodiments are only to illustrate the technical concept and characteristics of the present invention, and the purpose is to enable those who are familiar with the technology to understand the content of the present invention and implement it accordingly, and cannot limit the protection scope of the present invention. . All equivalent changes or modifications made according to the spirit of the present invention should be included within the protection scope of the present invention.

Claims (2)

1. The phenolic glycoside compound in the moringa seeds is characterized by comprising the following specific compounds, and the structure of the phenolic glycoside compound is as follows:

2. the phenolic glycoside compound of claim 1 and its use in preparing medicine and health product for treating diabetes, depression and senile dementia.

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