Crocins compounds and uses thereof

Abstract

Provided are a series of crocins compounds and related pharmacological applications thereof in prevention and treatment of Alzheimer's disease. The series of crocins compounds are obtained by taking the Chinese herb, namely Gardenia jasminoides Ellis, as a raw material and separating same by means of various methods. The compounds playing a role in preventing oxidative injury caused by hydrogen peroxide (H.sub.2O.sub.2) and excitatory amino acid injury caused by L-glutamic acid are screened out by in-vitro cell experiments. The results show that the crocins compounds have good effect in preventing cell injury caused by H.sub.2O.sub.2 and L-glutamic acid. The compounds have good effect in preventing and treating Alzheimer's disease due to the fact that significant rise of oxidative stress and excitatory amino acid is a key factor for nerve injury in Alzheimer's disease, and have broad development and application prospect.

Claims

1. A method for inhibiting or treating Alzheimer's disease in a subject, which comprises administering an effective amount of a crocin-like compound or a pharmaceutically acceptable salt thereof to the subject, wherein the crocin-like compound is selected from ##STR00055## and pharmaceutically acceptable salts thereof.

2. The method according to claim 1, wherein the compound is ##STR00056##

3. The method according to claim 1, wherein the compound is ##STR00057##

Description

DETAILED DESCRIPTION OF EMBODIMENTS

(1) Inventors of the present disclosure extracted a series of novel crocin-like compounds represented by a general formula (I) from a Chinese herb Gardenia jasminoides Ellis by multiple chemical means, and upon identification through multiple spectroscopic methods and cell experiments, it was proved that these crocin-like compounds have excellent protection effects on a central nervous system.

(2) ##STR00014##

(3) Various substituents are defined as in the preceding.

(4) In a first group of preferred compounds, a position 13-14 is of trans E configuration, R.sub.1 represents glucosyl group, and R.sub.2 represents a quinic acid group.

(5) In the above, when R.sub.1 represents Glc-(1.fwdarw.6)-Glc (gentiobiosyl) group, and R.sub.2 represents a 3-O-caffeoylquinic acid-4-oxy group, a new compound neocrocin B is obtained, referred to as GJ-1 for short.

(6) ##STR00015##

(7) In the above, when R.sub.1 represents gentiobiosyl group, and R.sub.2 represents a 3-O-caffeoylquinic acid-5-oxy group, a new compound neocrocin C is obtained, referred to as GJ-2 for short.

(8) ##STR00016##

(9) In a second group of preferred compounds, a position 13-14 is of cis Z configuration, R.sub.1 represents H, glucosyl group or a quinic acid group, and R.sub.2 represents glucosyl group or a quinic acid group.

(10) In the above, when R.sub.1 represents gentiobiosyl group, and R.sub.2 represents a 3-O-caffeoylquinic acid-4-oxy group, a new compound neocrocin D is obtained, referred to as GJ-3 for short.

(11) ##STR00017##

(12) In the above, when R.sub.1 represents a 3-O-caffeoylquinic acid-4-oxy group, and R.sub.2 represents gentiobiosyl group, a new compound neocrocin E is obtained, referred to as GJ-4 for short.

(13) ##STR00018##

(14) In the above, when R.sub.1 represents H, and R.sub.2 represents gentiobiosyl group, a new compound named as 13Z-crocetin-8′-O-β-D-gentiobioside is obtained, referred to as GJ-5 for short.

(15) ##STR00019##

(16) In a third group of preferred compounds, a position 13-14 is of trans E configuration, R.sub.1 represents glucosyl group, and R.sub.2 represents glucosyl group or H.

(17) In the above, when R.sub.1 represents a 6-O-trans-sinapoyl gentiobiosyl group, and R.sub.2 represents gentiobiosyl group, a new compound neocrocin G is obtained, referred to as GJ-6 for short.

(18) ##STR00020##

(19) In the above, when R.sub.1 represents a 6-O-trans-sinapoyl gentiobiosyl group, and R.sub.2 represents H, a new compound neocrocin F is obtained, referred to as GJ-7 for short.

(20) ##STR00021##

(21) In a fourth group of preferred compounds, a position 13-14 is of trans E configuration, R.sub.1 represents glucosyl group, and R.sub.2 represents CH.sub.2CH.sub.3.

(22) In the above, when R.sub.1 represents a gentiobiosyl group, and R.sub.2 represents CH.sub.2CH.sub.3, a new compound neocrocin H is obtained, referred to as GJ-8 for short.

(23) ##STR00022##

(24) A fifth group of preferred compound is neocrocin I, referred to as GJ-9 for short.

(25) ##STR00023##

(26) In a sixth group of preferred compounds, position 13-14 is of trans E configuration, R.sub.1 represents glucosyl group or xylosyl group, and R.sub.2 represents H.

(27) In the above, when R.sub.1 represents H, and R.sub.2 represents xylosyl-(1.fwdarw.6)-glucosyl group, a new compound neocrocin J is obtained, referred to as GJ-10 for short.

(28) ##STR00024##

(29) Besides, compounds listed in following Table 1 within the range of the general formula (I) are known.

(30) TABLE-US-00001 TABLE 1 Abbreviation Chemical formula structure GJ-11 GJ-12 GJ-13 GJ-14 GJ-15 GJ-16 0 GJ-17 GJ-18 GJ-19 GJ-20

EXAMPLES

Example 1: Extraction and Separation of Ingredients of Crocins in Gardenia jasminoides Ellis

(31) 40.0 Kg of dry and mature fruits of Gardenia jasminoides Ellis were taken, smashed in a suitable manner, and extracted under heating and reflux 3 times with 4 times amount of 60% ethanol, 2 hours at each time. Extract solutions were combined, and a solvent was removed under a reduced pressure, to obtain 6.2 kg of a total extract of Gardenia jasminoides Ellis (with a yield of 15.5%). The extract was dissolved in a suitable amount of water, and centrifuged, a supernatant was subjected to macroporous resin open column chromatography (20.0×90 cm), and gradient elution subsequently with water, 30% ethanol, 50% ethanol, 70% ethanol, and 95% ethanol at 4 times a column bed volume. And eluates in various fractions were collected, and solvents were recovered respectively under a reduced pressure, to obtain about 4.5 kg of a combined fraction from water elution and 30% ethanol elution, 710.0 g of fraction from 50% ethanol elution, 150.0 g of fraction from 70% ethanol elution, and 112.0 g of fraction from 95% ethanol elution, wherein the fraction from 70% ethanol elution is just an active fraction of crocins of Gardenia jasminoides Ellis.

(32) 150 g of fraction from 70% ethanol elution was subjected to silica gel column chromatography (φ7×60 cm), and gradient elution with 99:1-6:4:0.8 chloroform-methanol-water, to separate out compounds GJ-11 (49.1 mg), GJ-12 (136.5 mg), and GJ-13 (7.0 g), respectively.

(33) A silica gel sub-fraction Fr. 9 was subjected to ODS column chromatography, and gradient elution with 30%-70% methanol-water to separate out a compound GJ-19 (545.1 mg), subjected to HPLC and elution with 60% methanol-water to obtain a compound GJ-20 (265.7 mg), subjected to HPLC and elution with 68% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-1 (120.0 mg) and 104.8 mg of a mixture of GJ-3 and GJ-4 (mixed by 1:2). And further, compounds GJ-3 and GJ-4 were separated using water (0.3% TEAA):acetonitrile=55:45.

(34) A silica gel sub-fraction Fr. 7 was subjected to ODS column chromatography and gradient elution with 40%-80% methanol-water to separate out a compound GJ-11 (16.0 mg), subjected to HPLC and elution with 55% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-2 (6.3 mg), subjected to HPLC and elution with 42% acetonitrile-acid water solution (0.1% acetic acid) to obtain a compound GJ-13 (8.0 mg) and a compound GJ-5 (16.0 mg).

(35) A silica gel sub-fraction Fr. 8 was subjected to ODS column chromatography and gradient elution with 55%-65% methanol-water to separate out a compound 16 (143.7 mg), subjected to HPLC and elution with 55% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-6 (59.1 mg) and GJ-2 (21.9 mg), subjected to HPLC and elution with 68% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-1 (400.9 mg), subjected to HPLC and elution with 32% acetonitrile-acid water solution (0.1% acetic acid) to obtain a compound GJ-17 (1.8 mg) and GJ-18 (3.5 mg).

(36) A silica gel sub-fraction Fr. 6 was subjected to ODS column chromatography and gradient elution with 50%-90% methanol-water to separate out a compound GJ-15 (315.7 mg), subjected to HPLC and elution with 60% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-10 (10.0 mg), subjected to HPLC and elution with 65% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-9 (2.0 mg) and GJ-7 (66.2 mg), subjected to HPLC and elution with 70% methanol-acid water solution (0.1% acetic acid) to obtain a compound GJ-8 (10.0 mg).

(37) Structure information of the compounds obtained is listed in Table 2.

(38) TABLE-US-00002 TABLE 2 Serial NO No. Structure  1* GJ-1  2* GJ-2  3* GJ-3  4* GJ-4  5* GJ-5  6* GJ-6 0  7* GJ-7  8* GJ-8  9* GJ-9 10* GJ-10 11 GJ-11 12 GJ-12 13 GJ-13 14 GJ-14 15 GJ-15 16 GJ-16 0 17 GJ-17 18 GJ-18 19 GJ-19 20 GJ-20

(39) Physical and chemical data of the compounds obtained are as follows:

(40) Compound GJ-1: red amorphous powder; ESI-MS (positive): m/z 1011[M+Na].sup.+; HR-ESI-MS: m/z 989.3642[M+H].sup.+ (calcd for C.sub.48H.sub.61O.sub.22, 989.3654), confirming a molecular formula of the compound GJ-1 as C.sub.48H.sub.60O.sub.22; UV(MeOH)λ.sub.max(log ε): 433 (5.32), 458 (5.28), 331 (4.68), 253 (4.52)nm; IR(KBr)v.sub.max 968, 1061, 1224, 1268, 1576, 1610, 1694, 2920, 3401 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(41) Compound GJ-2: red amorphous powder; ESI-MS (positive): m/z 1011[M+Na].sup.+; HR-ESI-MS: m/z 989.3646[M+H].sup.+ (calcd for C.sub.48H.sub.61O.sub.22, 989.3654), confirming a molecular formula of the compound GJ-2 as C.sub.48H.sub.60O.sub.22; UV(MeOH)λ.sub.max(log ε): 431 (4.63), 457 (4.56), 331 (4.12), 249 (3.85); IR(KBr)v.sub.max 1064, 1230, 1279, 1602, 1698, 2921, 3417 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6 150 MHz).

(42) Compound GJ-3: red amorphous powder; ESI-MS (positive): m/z 1011[M+Na].sup.+; HR-ESI-MS: m/z 1011.3471[M+Na].sup.+ (calcd for C.sub.48H.sub.60O.sub.22Na, 1011.3474), confirming a molecular formula of the compound GJ-3 as C.sub.48H.sub.60O.sub.22; UV(MeOH)λ.sub.max(log ε): 429 (5.04), 453 (4.99), 324 (4.68), 251 (4.04)nm; IR(KBr)v.sub.max 969, 1062, 1229, 1277, 1607, 1693, 2920, 3368 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(43) Compound GJ-4: red amorphous powder; ESI-MS (positive): m/z 1011[M+Na].sup.+; HR-ESI-MS: m/z 1011.3471 [M+Na].sup.+ (calcd for C.sub.48H.sub.60O.sub.22Na, 1011.3474), confirming a molecular formula of the compound GJ-4 as C.sub.48H.sub.60O.sub.22; UV(MeOH)λ.sub.max(log ε): 429 (5.04), 453 (4.99), 324 (4.68), 251 (4.04)nm; IR(KBr)v.sub.max 969, 1062, 1229, 1277, 1607, 1693, 2920, 3368 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(44) Compound GJ-5: red amorphous powder, ESI-MS (positive): m/z 675[M+Na].sup.+, m/z 1327 [2M+Na].sup.+, inferring a molecular weight of the compound GJ-14 as 652; HR-ESI-MS: 675.2617 [M+Na.sup.+] (a calculated value is 675.2629), determining a molecular formula of the compound GJ-14 as C.sub.32H.sub.44O.sub.14. See Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(45) Compound GJ-6: red amorphous powder; ESI-MS (positive): m/z 1205 [M+Na].sup.+; HR-ESI-MS: m/z 1183.4479 [M+H].sup.+ (calcd for C.sub.55H.sub.75O.sub.28, 1183.4445), confirming a molecular formula of the compound GJ-6 as C.sub.55H.sub.74O.sub.28; UV(MeOH)λ.sub.max(log ε): 434 (5.22), 459 (5.17), 330 (4.78), 242 (4.65); IR(KBr)v.sub.max 1059, 1119, 1225, 1273, 1610, 1701, 2920, 3385 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(46) Compound GJ-7: red amorphous powder; ESI-MS (positive): m/z 881 [M+Na].sup.+; HR-ESI-MS: m/z 881.3188 [M+Na].sup.+ (calcd for C.sub.43H.sub.54O.sub.18Na, 881.3208), confirming a molecular formula of the compound GJ-7 as C.sub.43H.sub.54O.sub.18; UV(MeOH)λ.sub.max(log ε): 430 (5.33), 454 (5.28), 326 (4.80), 242 (4.78); IR(KBr)v.sub.max 972, 1069, 1179, 1227, 1284, 1610, 1697, 2922, 3391 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(47) Compound GJ-8: red amorphous powder; ESI-MS (positive): m/z 703 [M+Na].sup.+; HR-ESI-MS: m/z 703.2904 [M+Na].sup.+ (calcd for C.sub.34H.sub.48O.sub.14Na, 703.2942), confirming a molecular formula of the compound GJ-8 as C.sub.34H.sub.48O.sub.14; UV(MeOH)λ.sub.max(log ε): 430 (4.64), 456 (4.59), 322 (3.84), 257 (3.95); IR(KBr)v.sub.max 1074, 1229, 1697, 2925, 3400 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(48) Compound GJ-9: red amorphous powder; ESI-MS (positive): m/z 659 [M+Na].sup.+; HR-ESI-MS: m/z 659.2657 [M+Na].sup.+ (calcd for C.sub.32H.sub.44O.sub.13Na, 659.2680), confirming a molecular formula of the compound GJ-9 as C.sub.32H.sub.44O.sub.13; UV(MeOH)λ.sub.max(log ε): 438 (4.63), 462 (4.60), 328 (3.90), 261 (3.94); IR(KBr)v.sub.max 1071, 1515, 1694, 2921, 3277 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(49) Compound GJ-10: red amorphous powder; ESI-MS (positive): m/z 645 [M+Na].sup.+; HR-ESI-MS: m/z 645.2519 [M+Na].sup.+ (calcd for C.sub.31H.sub.42O.sub.13Na, 645.2523), confirming a molecular formula of the compound GJ-10 as C.sub.31H.sub.42O.sub.13; UV(MeOH)λ.sub.max(log ε): 428 (4.56), 453 (4.50), 320 (4.11), 257 (4.10); IR(KBr)v.sub.max 1072, 1230, 1700, 2924, 3416 cm.sup.−1; see Table 3 for .sup.13C NMR (DMSO-d.sub.6, 150 MHz).

(50) TABLE-US-00003 TABLE 3 NO. Pos. 1 2 3 4 5 6 7 8 9 10  8 166.2 166.2 166.2 166.9 169.3 166.2 166.2 166.2 166.2 166.2  8′ 167.0 167.1 166.9 166.2 166.2 166.2 169.1 167.4 194.5 169.3  9 125.4 125.2 125.4 126.8 128.1 125.3 125.2 125.2 125.4 125.1  9′ 126.1 127.1 125.9 125.1 125.0 125.3 127.0 126.2 136.5 127.3 10 140.0 139.9 140.0 139.0 137.9 139.9 139.9 139.9 139.9 139.9 10′ 139.0 138.3 139.0 140.0 140.0 139.9 138.0 138.4 149.2 137.9 11 124.0.sup.a 123.8 125.2 125.2 125.5 123.9 123.8 123.8 124.1 123.7 11′ 123.9.sup.a 124.2 123.7 123.6 123.5 123.9 124.2 124.0 123.9 124.3 12 144.7 144.7 136.8 135.6 135.3 144.6 144.6 144.6 144.5 144.7 12′ 144.0 143.4 144.1 144.8 144.7 144.6 143.3 143.9 145.7 143.2 13 136.9.sup.a 137.0 135.1 135.0 135.2 136.9 136.9 136.8.sup.a 136.8 137.0 13′ 136.8.sup.a 136.7 136.4 136.4 136.2 136.9 136.6 136.9.sup.a 137.3 136.6 14 136.1 136.0 134.2 134.2 133.7 136.0.sup.a 136.0 136.0 135.9 136.0 14′ 135.7 135.3 135.9 136.0 135.9 135.9.sup.a 135.3 135.6 137.0 135.2 15 132.1 132.1 130.9 130.9 131.1 132.0 132.0 132.0 131.9 132.1 15′ 131.9 131.7 130.7 130.7 130.3 132.0 131.6 131.7 132.6 131.5 19 12.7 12.7 12.7 12.7 12.8 12.7 12.7 12.7 12.7 12.7 19′ 12.8 12.8 12.8 12.8 12.7 12.7 12.8 12.8 9.4 12.9 20 12.6 12.5 20.0 20.1 20.1 12.5 12.6 12.6 12.6 12.5 20′ 12.6 12.6 12.5 12.5 12.5 12.6 12.5 12.5 12.5 12.6  1″ 73.6 72.8 73.6 73.6 166.7 166.7 60.1  2″ 37.4 35.1 37.0 37.0 114.8 114.8 14.2  3″ 67.5 70.9 67.7 67.7 145.5 145.5  4″ 74.2 68.5 73.4 73.4 124.4 124.4  5″ 66.3 70.9 66.1 66.1 106.3 106.3  6″ 37.6 35.1 37.6 37.6 148.0 148.0  7″ 174.8 172.1 174.7 174.7 138.3 138.3  8″ 148.0 148.0  9″ 106.3 106.3  6″, 56.1 56.1  8″-OCH.sub.3  1′″ 165.6 165.7 165.5 165.5  2′″ 113.6 114.3 113.6 113.6  3′″ 145.6 145.0 145.6 145.6  4′″ 125.2 125.6 125.4 125.4  5′″ 114.9 114.8 114.9 114.9  6′″ 145.6 145.6 145.6 145.6  7′″ 148.5 148.4 148.5 148.5  8′″ 115.7 115.8 115.7 115.7  9′″ 121.6 121.3 121.5 121.5 8-Gen 8-Gen 8-Gen 8′-Gen 8′-Gen 8-Gen 8-Gen 8-Gen 8-Gen  1 94.5 94.5 94.5 94.5 94.5 94.5 94.5 94.5 94.5 94.5  2 72.5 72.4 72.5 72.5 72.4 72.5 72.5 72.5 72.4 72.4  3 76.3 76.2 76.3 76.3 76.2 76.2 76.2 76.3 76.2 76.2  4 69.2 69.2 69.2 69.2 69.2 69.2 69.2 69.2 69.2 69.3  5 76.3 76.3 76.3 76.3 76.3 76.2 76.2 76.3 76.3 76.4  6 67.9 67.9 67.9 67.9 67.9 68.0 68.0 67.9 67.9 68.0  1′ 103.1 103.1 103.1 103.1 103.1 103.0 103.0 103.1 103.1 103.7  2′ 73.5 73.4 73.5 73.5 73.5 73.4 73.4 73.5 73.4 73.3  3′ 76.8 76.8 76.8 76.8 76.8 76.5 76.5 76.8 76.7 76.6  4′ 70.0 70.0 70.0 70.0 70.0 69.8 69.8 70.0 69.9 69.5  5′ 76.9 76.9 76.9 76.9 76.9 73.8 73.8 76.9 76.9 65.7  6′ 61.0 61.0 61.0 61.0 61.0 63.5 63.5 61.0 61.0 8′-Gen  1 94.5  2 72.5  3 76.3  4 69.2  5 76.3  6 67.9  1′ 103.1  2′ 73.5  3′ 76.8  4′ 70.0  5′ 76.9  6′ 61.0 .sup.ameans signals could be interchangeable with the corresponding position in one compound

Example 2: Neuroprotection Effect of Monomers of Crocins of Gardenia jasminoides Ellis in SH-SY5Y Cell Damage Models Caused by H.SUB.2.O.SUB.2 .and L-Glutamic Acid

(51) 2.1 Method for Culturing SH-SY5Y Nerve Cells

(52) The SH-SY5Y nerve cells were cultured in a DMEM culture medium (containing 5% of fetal calf serum in volume fraction) in an incubator containing 5% CO.sub.2 at 37° C., while subculturing every 3 to 4 days. Cells in a logarithmic phase were selected for experiments.

(53) 2.2 Method for Screening with Hydrogen Peroxide-Damaged Models

(54) SH-SY5Y cells were inoculated at a concentration of 5×10.sup.3 in a 96-well plate, and then cultured for 24 h, and 100 μL of a liquid culture medium containing H.sub.2O.sub.2 and a crocin-like compound was added to the 96-well plate, such that a final concentration of H.sub.2O.sub.2 was 400 μM and a final concentration of the crocin-like compound was 10 μM, 1 μM, 0.1 μM, with each concentration in triplicate, followed by culturing for 24 h. After 24 h, a supernatant was sucked out and discarded, and to each well was added 100 μL of MTT (0.5 mg/mL), followed by incubation for 4 h. A supernatant was sucked out and discarded, to each well was added 150 μL of DMSO, followed by shaking for 10 min, and a wavelength of 570 nm was selected to measure a value of absorbancy on a microplate reader.sup.[12]. (Effective rate %=(OD.sub.crocin-like compound−OD.sub.model)/(OD.sub.control−OD.sub.model)*100). See Table 4 for screening results.

(55) 2.3 Method for Screening with L-Glutamic Acid-Damaged Models

(56) SH-SY5Y cells were inoculated at a concentration of 5×10.sup.3 in a 96-well plate, and then cultured for 24 h, and 100 μL of a liquid culture medium containing L-glutamic acid and a crocin-like compound was added to a 96-well plate, such that a final concentration of L-glutamic acid was 160 mM and a final medication concentration of the crocin-like compound was 10 μM, 1 μM, 0.1 μM, with each concentration in triplicate, followed by culturing for 24 h. After 24 h, a supernatant was sucked out and discarded, to each well was added 100 μL of MTT (0.5 mg/mL), followed by incubation for 4 h. A supernatant was sucked out and discarded, to each well was added 150 μL of DMSO, followed by shaking for 10 min, and a wavelength of 570 nm was selected to measure a value of absorbancy on a microplate reader.sup.[13]. (Effective rate %=(OD.sub.crocin-like compound−OD.sub.model)/(OD.sub.control−OD.sub.model)*100). See Table 5 for results.

(57) TABLE-US-00004 TABLE 4 Concentration of Sequence Serial No. of Crocin-like Survival Rate of Cells with No. Compounds Cell Model Compound (mol/L) Crocin-like Compound  1 GJ-1 H.sub.2O.sub.2SY5Y 10.sup.−5 10.39 ± 4.85  Damage 10.sup.−6 0.17 ± 0.29 Model 10.sup.−7 0.00 ± 0.00  2 GJ -2 H.sub.2O.sub.2SY5Y 10.sup.−5 8.82 ± 6.57 Damage 10.sup.−6 0.00 ± 0.35 Model 10.sup.−7 0.00 ± 0.00  3 GJ-3/GJ-4 H.sub.2O.sub.2SY5Y 10.sup.−5 5.49 ± 5.57 Damage 10.sup.−6 0.00 ± 0.00 Model 10.sup.−7 0.25 ± 0.08  5 GJ-6 H.sub.2O.sub.2SY5Y 10.sup.−5   36.38 ± 5.27*** Damage 10.sup.−6  26.02 ± 3.62** Model 10.sup.−7  14.56 ± 11.22*  6 GJ-7 H.sub.2O.sub.2SY5Y 10.sup.−5 18.60 ± 3.43* Damage 10.sup.−6 6.83 ± 3.63 Model 10.sup.−7 2.42 ± 4.19  7 GJ-8 H.sub.2O.sub.2SY5Y 10.sup.−5 36.60 ± 3.81* Damage 10.sup.−6 5.09 ± 4.50 Model 10.sup.−7 2.28 ± 3.95  8 GJ-9 H.sub.2O.sub.2SY5Y 10.sup.−5 9.43 ± 1.47 Damage 10.sup.−6 0.67 ± 1.13 Model 10.sup.−7 0.00 ± 0.00  9 GJ-10 H.sub.2O.sub.2SY5Y 10.sup.−5  33.92 ± 2.30*  Damage 10.sup.−6 16.98 ± 3.74  Model 10.sup.−7 6.28 ± 5.68 10 GJ-11 H.sub.2O.sub.2SY5Y 10.sup.−5 2.20 ± 3.81 Damage 10.sup.−6 4.29 ± 4.42 Model 10.sup.−7 2.27 ± 2.82 11 GJ-12 H.sub.2O.sub.2SY5Y 10.sup.−5 0.00 ± 0.00 Damage 10.sup.−6 0.00 ± 0.00 Model 10.sup.−7 0.00 ± 0.00 12 GJ-15 H.sub.2O.sub.2SY5Y 10.sup.−5 0.00 ± 0.00 Damage 10.sup.−6 0.00 ± 0.00 Model 10.sup.−7 0.00 ± 0.00 13 GJ-16 H.sub.2O.sub.2SY5Y 10.sup.−5 0.00 ± 0.00 Damage 10.sup.−6 0.00 ± 0.00 Model 10.sup.−7 0.00 ± 0.00 *P < 0.1, **P < 0.05, ***P < 0.01.

(58) TABLE-US-00005 TABLE 5 Serial Concentration of Survival Rate Sequence No. of Crocin-like of Cells with No. Compounds Cell Model Compound (mol/L) Crocin-like Compound  1 GJ-1 L-Glu SY5Y 10.sup.−5  40.03 ± 3.91** Damage Model 10.sup.−6 27.63 ± 5.36* 10.sup.−7 9.89 ± 2.93  2 GJ-2 L-Glu SY5Y 10.sup.−5 16.29 ± 14.12 Damage Model 10.sup.−6 7.69 ± 9.01 10.sup.−7  6.68 ± 11.57  3 GJ-3/GJ-4 L-Glu SY5Y 10.sup.−5  12.95 ± 4.01** Damage Model 10.sup.−6 2.78 ± 3.24 10.sup.−7 1.51 ± 2.62  5 GJ-6 L-Glu SY5Y 10.sup.−5  55.02 ± 0.60** Damage Model 10.sup.−6  48.75 ± 9.49** 10.sup.−7  24.51 ± 13.70*  6 GJ-7 L-Glu SY5Y 10.sup.−5  74.93 ± 15.36* Damage Model 10.sup.−6   57.05 ± 10.87** 10.sup.−7  42.60 ± 5.40**  7 GJ-8 L-Glu SYSY 10.sup.−5 41.28 ± 7.52* Damage Model 10.sup.−6 7.39 ± 7.08 10.sup.−7 0.00 ± 0.00  8 GJ-9 L-Glu SY5Y 10.sup.−5 46.12 ± 8.45* Damage Model 10.sup.−6 38.02 ± 5.15* 10.sup.−7 18.26 ± 5.02   9 GJ-10 L-Glu SY5Y 10.sup.−5  35.20 ± 13.61* Damage Model 10.sup.−6  33.15 ± 12.55* 10.sup.−7 18.26 ± 5.02  10 GJ-11 L-Glu SY5Y 10.sup.−5 4.80 ± 1.28 Damage Model 10.sup.−6 7.02 ± 4.12 10.sup.−7 9.09 ± 4.62 11 GJ-12 L-Glu SY5Y 10.sup.−5 0.00 ± 0.00 Damage Model 10.sup.−6 0.00 ± 0.00 10.sup.−7 0.00 ± 0.00 12 GJ-15 L-Glu SY5Y 10.sup.−5 0.00 ± 0.00 Damage 10.sup.−6 0.00 ± 0.00 Model 10.sup.−7 0.00 ± 0.00 13 GJ-16 L-Glu SY5Y 10.sup.−5 0.00 ± 0.00 Damage Model 10.sup.−6 0.00 ± 0.00 10.sup.−7 0.00 ± 0.00 14 GJ-17/GJ-18 L-Glu SY5Y 10.sup.−5 0.00 ± 0.00 Damage Model 10.sup.−6 0.00 ± 0.00 10.sup.−7 0.00 ± 0.00 *P < 0.1, **P < 0.05, ***P < 0.01.

(59) Experiment results indicate: on the SY5Y cell damage models induced by H.sub.2O.sub.2, all of the compounds GJ-1 to GJ-10 manifested good protection effects, with compounds GJ-6, GJ-10, and GJ-8 manifesting more excellent protection effects; on the SY5Y cell damage models induced by L-glutamic acid, all of the compounds GJ-1 to GJ-10 also manifested good protection effects, with compounds GJ-1, GJ-6, GJ-7, GJ-10, GJ-9, and GJ-8 manifesting more excellent protection effects. It also can be obtained from the above experiment results that various compounds had different effectiveness in different damage models, which may be resulted from that a mechanism of H.sub.2O.sub.2-induced damage is oxidative stress, while a mechanism of L-glutamic acid-induced damage is excitotoxicity damage caused by an excitatory amino acid, and thus the compounds exert the protective effects against damage through different mechanisms.