CN114773200B

CN114773200B - Vulgarisin diterpenoid compound, extraction method and application

Info

Publication number: CN114773200B
Application number: CN202210558839.1A
Authority: CN
Inventors: 马风伟; 潘卫东; 娄华勇; 李金玉; 刘翰飞; 邓青芳; 张妮
Original assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Current assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Priority date: 2022-05-21
Filing date: 2022-05-21
Publication date: 2023-07-18
Anticipated expiration: 2042-05-21
Also published as: CN114773200A

Abstract

The invention relates to the technical field of natural medicines, in particular to a vulgarisin diterpenoid compound, an extraction method and application. The structural general formula of the vulgarisin diterpenoid compound is shown as I. The vulgarisin diterpenoid compounds have certain neuron protective activity in preliminary neuroprotective activity screening discovery, can be used for treating cerebral diseases such as cerebral apoplexy, senile dementia and the like and preventing neuroprotection, and mainly has the effects of preventing and treating neuronal cell death and improving survival rate and activity.

Description

Vulgarisin diterpenoid compound, extraction method and application

Technical Field

The invention relates to the technical field of natural medicines, in particular to a vulgarisin diterpenoid compound, an extraction method and application.

Background

The vulgarisin diterpenoid compounds are drugs with pharmacodynamic activity. The application finds that the vulgarisin diterpenoid compound can be extracted from the selfheal, and the obtained vulgarisin diterpenoid compound belongs to a brand new compound and has corresponding medicinal value.

Disclosure of Invention

In view of the above-described drawbacks of the prior art, an object of the present invention is to provide a vulgarisin-type diterpenoid compound; meanwhile, the invention also provides a method for extracting the vulgarisin diterpenoid compounds from the selfheal; in addition, the invention also provides application of the vulgarisin diterpenoid compound in preparation of medicines with neuron protective activity.

To achieve the above-mentioned objects and other related objects,

in a first aspect of the present invention, there is provided a vulgarisin type diterpenoid compound, wherein the structural general formula of the vulgarisin type diterpenoid compound is shown as I:

r1, R5 and R7 are independently selected from H or alkyl;

r2, R3 and R4 are independently selected from hydroxyl,One of the following;

and R6 is H or hydroxyl.

In one embodiment of the present invention, R1, R5 and R7 are independently selected from alkyl groups having 1 to 6 carbon atoms.

In one embodiment of the present invention, R1 and R7 are both straight-chain alkyl groups having 1 to 3 carbon atoms, and R5 is branched-chain alkyl groups having 3 to 5 carbon atoms; .

In an embodiment of the invention, R1 and R7 are both methyl, R5 is isopropyl, and R6 is hydroxy.

In one embodiment of the invention, R2 or R3 is hydroxy.

In one embodiment of the present invention, R2 is hydroxy and R3 isOne of them.

In one embodiment of the present invention, R3 is hydroxy and R2 isOne of them.

In one embodiment of the present invention, the vulgarisin-type diterpenoid compound is

In a second aspect of the present invention, there is provided a method for extracting vulgarisin type diterpenoid compounds from Prunella vulgaris, comprising the steps of:

extracting Prunellae Spica with ethanol to obtain extract;

extracting the extract with petroleum ether to obtain a pre-extract;

gradually eluting the pre-extract by adopting a petroleum ether-ethyl acetate system according to the petroleum ether ratio to obtain a crude extract of the vulgarisin diterpenoid compound;

eluting the crude extract of the vulgarisin diterpenoid compound by adopting a methanol-water system according to the gradual increase of the ratio of methanol to obtain the vulgarisin diterpenoid compound.

In one embodiment of the invention, the volume ratio of the pre-extract to the petroleum ether-ethyl acetate is 100:0- & gt (85-95): (5-15) → (65-75): (25-35) → (45-55): (45-55) → (15-25): gradient elution is carried out from (75-85) to 0:100, and Fr.A, fr.B, fr.C, fr.D, fr.E and Fr.F are obtained in sequence;

Fr.D is sequentially (35-45) according to the volume ratio of methanol to water: (55-65) → (45-55): (45-55) → (55-65): (35-45) → (65-75): (25-35) → (75-85): (15-25) → (85-95): (5-15) to 100:0 to obtain Fr.D1, fr.D2, fr.D3, fr.D4, fr.D5, fr.D6 and Fr.D7 in sequence;

Fr.D5 is sequentially (35-45) according to the volume ratio of methanol to water: (55-65) → (55-65): (35-45) → (75-85): (15-25) → (85-95): (5-15) to 100:0 to obtain Fr.D5-1, fr.D5-2, fr.D5-3, fr.D5-4 and Fr.D5-5 in sequence;

Fr.D5-1 is mixed with methanol to water according to the volume ratio of (80-95): (5-20) eluting to obtain a compound 1, a compound 3, a compound 7 and a compound 11;

Fr.D5-2 is mixed with methanol to water according to the volume ratio of (70-90): (10-30) eluting to obtain a compound 2, a compound 8 and a compound 10;

Fr.D5-3 is mixed with methanol to water according to the volume ratio of (85-95): (5-15) eluting to obtain a compound 4, a compound 5, a compound 9 and a compound 12;

Fr.D5-4 is mixed according to the volume ratio of methanol to water of (90-100): (2-10) eluting to obtain a compound 6;

wherein the compounds 1 to 12 are in turn

In a third aspect, the present invention provides an application of the vulgarisin diterpenoid compound in preparation of a medicament with neuron protective activity.

In an embodiment of the invention, the active drug with neuron protection is a drug for treating cerebral apoplexy, a drug for treating senile dementia or a drug for treating neurodegenerative diseases.

As described above, the vulgarisin diterpenoid compound, the extraction method and the application of the invention have the following beneficial effects:

1. the invention obtains the vulgarisin diterpenoid compounds (compounds 1 to 6) from the whole plant of the selfheal plant for the first time. Their planar structure was determined by HR-ESI-MS, 1D and 2D NMR methods, and the absolute configuration of the 6 compounds was further determined by X-ray or circular dichroism.

2. The compounds 1-12 separated from the selfheal have certain neuron protective activity through preliminary neuroprotective activity screening, and can be used for treating cerebral diseases such as cerebral apoplexy, senile dementia and the like and preventing neuroprotection, mainly preventing and treating neuronal cell death and improving survival rate and activity.

Detailed Description

Further advantages and effects of the present invention will become apparent to those skilled in the art from the disclosure of the present invention, which is described by the following specific examples.

The structures of compounds 1-12 are shown below:

1. extraction and separation experiment part

Example 1

Extraction and separation of Compounds 1 to 12

Step one, crushing the whole herb of the selfheal after naturally air-drying (dry weight is about 20 kg), soaking in 60L of 95% ethanol, heating and reflux-extracting for two times, and concentrating the extract to obtain 3.3kg of extract.

Dispersing the extract in 4L of water, and sequentially extracting with petroleum ether, ethyl acetate and water saturated n-butanol with equal volume. Preliminary experiments prove that the petroleum ether part contains abundant vulgarisin diterpenoid compounds. Therefore, the basic research of the subsequent pharmacodynamic substances mainly focuses on the petroleum ether part of the selfheal.

Step three, petroleum ether section extract (417.0 g) is subjected to gradient elution by a petroleum ether-ethyl acetate system (100:0-90:10-70:30-50:50-20:80-0:100) through silica gel medium pressure column chromatography (200-300 meshes) to obtain 6 components (Fr.A-Fr.F), wherein the specific steps are shown in table 1:

elution system	Eluted fractions
		The volume ratio of petroleum ether to ethyl acetate is 100:0	Fr.A
The volume ratio of petroleum ether to ethyl acetate is 90:10	Fr.B
		The volume ratio of petroleum ether to ethyl acetate is 70:30	Fr.C
The volume ratio of petroleum ether to ethyl acetate is 50:50	Fr.D
		The volume ratio of petroleum ether to ethyl acetate is 20:80	Fr.E
The volume ratio of petroleum ether to ethyl acetate is 0:100	Fr.F

TABLE 1

Step four, combining TLC analysis and UPLC-ESI-Q-exact-MS analysis; component FrD (104.0 g) was subjected to MCI column chromatography with a methanol-water system as the mobile phase for gradient elution (40:60→50:50→60:40→70:30→80:20→90:10→pure methanol, v/v) to give 7 components (FrD-D7), as shown in Table 2:

elution system	Eluted fractions
		The volume ratio of methanol to water is 40:60	FrD1
The volume ratio of methanol to water is 50:50	FrD2
		The volume ratio of methanol to water is 60:40	FrD3
The volume ratio of methanol to water is 70:30	FrD4
		The volume ratio of methanol to water is 80:20	FrD5
The volume ratio of methanol to water is 90:10	FrD6
		The volume ratio of methanol to water is 100:0	FrD7

TABLE 2

Step five, the component FrD5 is subjected to gradient elution by polyamide column chromatography by taking a methanol-water system as a mobile phase (40:60-60:40-80:20-90:10-100:0, v/v) to obtain 5 components (FrD 5-1-FrD 5-5), wherein the specific steps are shown in the table 3:

elution system	Eluted fractions
		The volume ratio of methanol to water is 40:60	FrD5-1
The volume ratio of methanol to water is 60:40	FrD5-2
		The volume ratio of methanol to water is 80:20	FrD5-3
The volume ratio of methanol to water is 90:10	FrD5-4
		The volume ratio of methanol to water is 100:0	FrD5-5

TABLE 3 Table 3

Step six, fraction FrD5-1 (909.8 mg) was isolated by semi-preparative HPLC eluting with a methanol-water (87:13, v/v;2 mL/min) system to give compound 1 (Rt 18.5 min), compound 3 (Rt 19.6 min), compound 7 (Rt 20.3 min) and compound 11 (Rt 23.5 min).

Step seven, fraction FrD5-2 (493.0 mg) was isolated by semi-preparative HPLC (phenyl column) eluting isocratically with a methanol-water (80:20, v/v;2 mL/min) system to give compound 2 (Rt 25.5 min), compound 8 (Rt 27.3 min) and compound 10 (Rt 20.2 min).

Step eight, component FrD5-3 (726.3 mg) was purified by Sephadex LH-20 and then separated by semi-preparative HPLC eluting isocratically with a methanol-water (92:8, v/v;2 mL/min) system to give compound 4 (Rt 18.5 min), compound 5 (Rt 16.6 min), compound 9 (Rt 15.0 min) and compound 12 (Rt 19.9 min).

Step nine, component FrD5-4 (87.0 mg) was isolated by semi-preparative HPLC as compound 6 (Rt 14.2 min) by isocratic elution with a methanol-water (94:6, v/v;2 mL/min) system.

Summarizing the compound yields obtained by separation in steps six to nine, as shown in table 4:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	16.9	10.0	18.9	23.0	22.5	18.9
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	15.0	25.0	112.1	167.0	133.0	105.0

TABLE 4 Table 4

FIG. 1 is a diagram of Compound 1 ¹ HNMR spectra, FIG. 2 is a graph of Compound 1 ¹³ C NMR spectrum, FIG. 3 is HSQC spectrum of Compound 1, FIG. 4 is HMBC spectrum of Compound 1, FIG. 5 is Compound 1 ¹ H- ¹ H COSY spectra, fig. 6 is NOESY spectra of compound 1, and fig. 7 is a high resolution mass spectrum of compound 1.

FIG. 8 is a diagram of Compound 2 ¹ An H NMR spectrum, FIG. 9 is a chart of Compound 2 ¹³ C NMR spectrum, FIG. 10 is HSQC spectrum of Compound 2, FIG. 11 is HMBC spectrum of Compound 2, FIG. 12 is Compound 2 ¹ H- ¹ H COSY spectrum, fig. 13 is NOESY spectrum of compound 2, fig. 14 is high resolution mass spectrum of compound 2.

FIG. 15 is a diagram of Compound 3 ¹ HNMR spectra, FIG. 16 is a representation of Compound 3 ¹³ C NMR spectrum, FIG. 17 is HSQC spectrum of Compound 3, FIG. 18 is HMBC spectrum of Compound 3, FIG. 19 is Compound 3 ¹ H- ¹ H COSY spectra, fig. 20 is NOESY spectra of compound 3, and fig. 21 is a high resolution mass spectrum of compound 3.

FIG. 22 is a diagram of Compound 4 ¹ HNMR spectra, FIG. 23 is a plot of Compound 4 ¹³ C NMR spectrum, FIG. 24 is HSQC spectrum of Compound 4, FIG. 25 is HMBC spectrum of Compound 4, FIG. 26 is Compound 4 ¹ H- ¹ H COSY spectrum, FIG. 27 is NOESY spectrum of Compound 4Fig. 28 is a high resolution mass spectrum of compound 4.

FIG. 29 is a diagram of Compound 5 ¹ HNMR spectra, FIG. 30 is a diagram of Compound 5 ¹³ C NMR spectrum, FIG. 31 is HSQC spectrum of Compound 5, FIG. 32 is HMBC spectrum of Compound 5, FIG. 33 is Compound 5 ¹ H- ¹ H COSY spectrum, fig. 34 is NOESY spectrum of compound 5, fig. 35 is high resolution mass spectrum of compound 5.

FIG. 36 is a diagram of Compound 6 ¹ HNMR spectra, FIG. 37 is a representation of Compound 6 ¹³ C NMR spectrum, FIG. 38 is HSQC spectrum of Compound 6, FIG. 39 is HMBC spectrum of Compound 6, and FIG. 40 is Compound 6 ¹ H- ¹ H COSY spectra, fig. 41 is NOESY spectra of compound 6, and fig. 42 is high resolution mass spectra of compound 6.

FIG. 43 is a diagram showing the structure of X-ray single crystals of Compound 1, FIG. 44 is a diagram showing the circular dichroism spectrum of Compound 1, FIG. 45 is a diagram showing the circular dichroism spectrum of Compound 2, FIG. 46 is a diagram showing the circular dichroism spectrum of Compound 3, FIG. 47 is a diagram showing the circular dichroism spectrum of Compound 4, FIG. 48 is a diagram showing the circular dichroism spectrum of Compound 5, and FIG. 49 is a diagram showing the circular dichroism spectrum of Compound 6.

Example 2

Example 2 differs from example 1 in the elution conditions of step three, as shown in table 5:

elution system	Eluted fractions
		The volume ratio of petroleum ether to ethyl acetate is 100:0	Fr.A
The volume ratio of petroleum ether to ethyl acetate is 90:10	Fr.B
		The volume ratio of petroleum ether to ethyl acetate is 70:30	Fr.C
The volume ratio of petroleum ether to ethyl acetate is 49:51	Fr.D
		The volume ratio of petroleum ether to ethyl acetate is 20:80	Fr.E
The volume ratio of petroleum ether to ethyl acetate is 0:100	Fr.F

Table 5 the yields of the compounds of example 2 are shown in table 6:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	17.6	9.2	16.8	20.1	22.5	13.5
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	16.6	22.8	130.3	177.7	135.5	110.0

TABLE 6

Example 3

Example 3 differs from example 1 in the elution conditions of step three, as shown in table 7:

elution system	Eluted fractions
		The volume ratio of petroleum ether to ethyl acetate is 100:0	Fr.A
The volume ratio of petroleum ether to ethyl acetate is 90:10	Fr.B
		The volume ratio of petroleum ether to ethyl acetate is 70:30	Fr.C
The volume ratio of petroleum ether to ethyl acetate is 45:55	Fr.D
		The volume ratio of petroleum ether to ethyl acetate is 20:80	Fr.E
The volume ratio of petroleum ether to ethyl acetate is 0:100	Fr.F

Table 7 the yields of the compounds of example 3 are shown in table 8:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	16.6	11.4	18.1	22.1	22.2	17.7
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	15.6	28.9	130.0	118.8	166.6	150.5

TABLE 8

Example 4

Example 4 differs from example 1 in the elution conditions of step three, as shown in table 9:

elution system	Eluted fractions
		The volume ratio of petroleum ether to ethyl acetate is 100:0	Fr.A
The volume ratio of petroleum ether to ethyl acetate is 90:10	Fr.B
		The volume ratio of petroleum ether to ethyl acetate is 70:30	Fr.C
The volume ratio of petroleum ether to ethyl acetate is 55:45	Fr.D
		The volume ratio of petroleum ether to ethyl acetate is 20:80	Fr.E
The volume ratio of petroleum ether to ethyl acetate is 0:100	Fr.F

Table 9 the yields of the compounds of example 4 are shown in table 10:

table 10

Example 5

Example 5 differs from example 1 in the elution conditions of step four, as shown in table 11:

elution system	Eluted fractions
		The volume ratio of methanol to water is 40:60	FrD1
The volume ratio of methanol to water is 50:50	FrD2
		The volume ratio of methanol to water is 60:40	FrD3
The volume ratio of methanol to water is 70:30	FrD4
		The volume ratio of methanol to water is 78:22	FrD5
The volume ratio of methanol to water is 90:10	FrD6
		The volume ratio of methanol to water is 100:0	FrD7

Table 11 yields of the compounds of example 5 are shown in table 12:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	18.8	11.3	20.5	24.1	25.2	23.2
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	13.6	26.6	167.7	155.5	120.5	142.2

Table 12

Example 6

Example 6 differs from example 1 in the elution conditions of step four, as shown in table 13:

elution system	Eluted fractions
		The volume ratio of methanol to water is 40:60	FrD1
The volume ratio of methanol to water is 50:50	FrD2
		The volume ratio of methanol to water is 60:40	FrD3
The volume ratio of methanol to water is 70:30	FrD4
		The volume ratio of methanol to water is 82:18	FrD5
The volume ratio of methanol to water is 90:10	FrD6
		The volume ratio of methanol to water is 100:0	FrD7

Table 13 yields of the compounds of example 6 are shown in table 14:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	19.6	11.2	20.5	26.6	20.2	15.5
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	16.8	26.8	144.4	200.3	166.1	115.0

TABLE 14

Example 7

Example 7 differs from example 1 in the elution conditions of step four, as shown in table 15:

elution system	Eluted fractions
		The volume ratio of methanol to water is 35:65	FrD5-1
The volume ratio of methanol to water is 55:45	FrD5-2
		The volume ratio of methanol to water is 75:25	FrD5-3
The volume ratio of methanol to water is 90:10	FrD5-4
		The volume ratio of methanol to water is 100:0	FrD5-5

Table 15 yields of the compounds of example 7 are shown in table 16:

compounds of formula (I)	1	2	3	4	5	6
							Yield/mg	13.8	6.5	18.2	21.1	15.5	17.4
Compounds of formula (I)	7	8	9	10	11	12
							Yield/mg	21.5	25.6	132.3	158.8	146.2	100.1

Table 16

Example 8

Example 8 differs from example 1 in the elution conditions of step four, as shown in table 17:

elution system	Eluted fractions
		The volume ratio of methanol to water is 42:58	FrD5-1
The volume ratio of methanol to water is 62:38	FrD5-2
		The volume ratio of methanol to water is 82:18	FrD5-3
The volume ratio of methanol to water is 90:10	FrD5-4
		The volume ratio of methanol to water is 100:0	FrD5-5

Table 17 yields of the compounds of example 8 are shown in table 18:

TABLE 18

2. Biological Activity test

Screening of neuroprotective Activity of Compounds 1-12

1. Determination of cell viability Using CCK8

Cell culture: fetal brains were removed after C57BL/6 pregnant mice were sacrificed, vascular membranes and other tissues were removed to obtain cerebral cortex, digested with 0.5%o trypsin and 0.1KU DNaseI/mL, gently blown at 37℃for about 20min, blown once every 10min, and then digestion was stopped with DMEM containing 10% FBS. The cells obtained were centrifuged (1000 rpm,3 to 5 min), resuspended and counted, and the cells were plated at an appropriate density in 96-well plates using DMEM medium containing 10% fbs+1% streptavidin diabodies. The cell culture dishes and cell culture plates used were all pre-coated with poly-L-lysine. Culturing in incubator at 37deg.C for 4 hr, changing liquid to neuron culture medium, and continuously culturing in 5% CO ₂ Is a 37℃incubator. And then, the liquid is changed once every 3 days, and the 8 th neuron is the mature neuron, so that the subsequent treatment can be performed.

Preparing CCK8 working solution: according to 1:10 mixing CCK8 solution with culture medium for use. It is ready for use.

Incubation of CCK 8: the 96-well cell culture plate was removed from the 37℃cell incubator, the original medium was aspirated, and the prepared CCK8 working solution was added to the 96-well plate in an amount of 100. Mu.L/well. The 96-well plate is put back into the cell incubator again for incubation for 1-4 h (incubation time is from OD _450nm About 1.0).

OD450 nm assay: OD of each well was measured with a multifunctional microplate reader _450nm . Measurement of suctionThe wavelength was set at 450nm, the pre-shaking was measured for 10s for mixing, and the absorbance was measured. The effect of the compound on primary neuronal cell viability was examined.

2. Observation of compounds for neuroprotective Activity Using OGD/R

The above treated neuronal cells were subjected to oxygen glucose deprivation-reoxygenation complex carbohydrate injury, and then the neuroprotective activity of the compounds was observed. The specific operation is as follows:

OGD/R, the sugar-free DMEM is first saturated with a mixture of 95% N2-5% CO2 for 15-30 min and preheated at 37 ℃. Discarding original cell culture medium, washing with pre-heated oxygen-free sugarless culture medium for 3 times, adding appropriate amount of oxygen-free sugarless DMEM, and culturing in a three-gas incubator at 37deg.C (94% N) ₂ ，5％CO ₂ ，1％O ₂ ) Culturing. After 2h, the cells were replaced with the corresponding normal medium for further culture for 22h, and the subsequent CCK8 experiments were performed.

The absorbance at a wavelength of 450nm was measured and the cell viability was calculated. Cell viability = [ (treatment well absorbance-blank well absorbance)/(control well absorbance-blank well absorbance) ]x100%.

3. Neuroprotection

Compounds 1-12 were dissolved in DMSO and finally diluted to a fixed concentration of 10. Mu.M. The experiment adopts a neuronal cell to carry out an oxygen glucose deprivation-reoxygenation complex glucose injury (OGD/R) model, and the activity influence condition of the compounds 1-12 on the neuronal cell is evaluated.

The Ctrl group showed that the OGD/R operation was not performed, so that the cell activity was 100%.

DMSO is solvent.

The positive control drugs were: edaravone.

The neuroprotective cell activities (i.e., cell viability) of compounds 1-12, ctrl group, DMSO, positive control are shown in table 19:

TABLE 19

Through activity tests, the compounds 1-12 have obvious neuroprotective activity, and the individual compounds are equivalent to or better than the positive control drugs. The neuroprotective activity of compounds 1 to 12 of example 1 is shown in figure 50.

In summary, compounds 1-12 of the present invention all have significant neuroprotective activity, with individual compounds being comparable or superior to the positive control. Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.

The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.

Claims

1. A vulgarisin-type diterpenoid compound, characterized in that the vulgarisin-type diterpenoid compound has the following structure:

2. a method for extracting vulgarisin diterpenoid compounds from selfheal, which is characterized by comprising the following steps:

extracting Prunellae Spica with ethanol to obtain extract;

extracting the extract with petroleum ether to obtain a pre-extract;

the volume ratio of the pre-extract to the petroleum ether-ethyl acetate is sequentially 100:0- & gt (85-95): (5-15) → (65-75): (25-35) → (45-55): (45-55) → (15-25): gradient elution is carried out from (75-85) to 0:100, and Fr.A, fr.B, fr.C, fr.D, fr.E and Fr.F are obtained in sequence;

wherein the compound 1 isThe compound 2 is->The compound 3 is->The compound 4 is->The compound 5 is->The compound 6 is->The compound 7 is/>The compound 8 is->The compound 9 is->The compound 10 is->The compound 11 is->The compound 12 is->

3. Use of a vulgarisin-type diterpenoid compound according to claim 1 for the preparation of a medicament with neuronal protection activity.

4. A use according to claim 3, characterized in that: the medicine with neuron protective activity is used for treating cerebral apoplexy, senile dementia or neurodegenerative diseases.