Substituted alcohol schizonepeta lactone compound as well as preparation method and application thereof

Abstract

The invention relates to a substituted alcohol nepetalactone compound as well as a preparation method and application thereof, and relates to the technical field of medicinal anti-inflammatory analgesia. The substituted alcohol type nepeta lactone compound is 4, 7-dimethyl-5, 6, 7, 7-tetrahydro-cyclopentadienopyran-1-alcohol or a medicinal salt or a medicinal esterification derivative of the 4, 7-dimethyl-5, 6, 7, 7-tetrahydro-cyclopentadienopyran-1-alcohol, and the substituted alcohol type nepeta lactone compound is a substituted alcohol type nepeta lactone compound. A zebrafish inflammation model is utilized, it is found for the first time that the substituted alcohol nepetalactone has excellent anti-inflammatory effect and analgesic effect, the mRNA levels of TNF-alpha, NF-kB and LDHA of the zebrafish inflammation model are obviously reduced, the increase of the movement distance of zebrafish caused by glacial acetic acid pain is obviously reduced, and the substituted alcohol nepetalactone has a remarkable effect at low concentration. The compound has the characteristics of difficulty in generating resistance, safety, high efficiency and low cost, can become a novel anti-inflammatory and analgesic alternative medicine, and is expected to assist in inflammation treatment.

Description

Technical Field

[0001] The invention relates to the technical field of medicinal anti-inflammatory and analgesic drugs, and in particular to a substituted alcohol nepetalactone compound and a preparation method and application thereof. Background Art

[0002] Inflammation is a defensive response of the body to stimulation, manifested as redness, swelling, heat, pain and dysfunction. Inflammation may be infectious inflammation caused by infection, or it may be non-infectious inflammation not caused by infection. Normally, inflammation is beneficial and is an automatic defensive response of the human body; but sometimes, inflammation is also harmful, such as attacks on the body's own tissues and inflammation occurring in transparent tissues, which can cause damage to the body and cells. Pain is an unpleasant feeling and emotional sensation accompanied by actual or potential tissue damage. Inflammatory reactions and pain reactions usually occur together, and targeted drugs with multiple effects of anti-inflammatory and analgesia have not yet been developed and widely used, which brings great difficulties to clinical drug treatment.

[0003] Nepeta cataria L. is a perennial plant of the genus Nepeta in the family Lamiaceae. It is a traditional Chinese medicinal material, mainly used to treat trauma, cerebral hemorrhage, blood in the stool, hematemesis, stroke and other symptoms. Studies have reported that the main active ingredient of Nepeta is "nepetalactone", which is a general term for a class of compounds. Further studies have shown that "nepetalactone" has a variety of structures. However, so far, there has been no report that substituted alcohol nepetalactone has the effect of relieving inflammation and pain reactions. In view of this, the present invention provides a substituted alcohol nepetalactone compound and a preparation method and application thereof. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a substituted alcohol nepetalactone compound and a preparation method and application thereof.

[0005] The technical solution of the present invention to solve the above technical problems is as follows:

[0006] In the first aspect, a substituted alcohol nepetalactone compound is provided, wherein the substituted alcohol nepetalactone compound comprises a compound as shown in formula (I) or a pharmaceutically acceptable salt or pharmaceutically acceptable esterified derivative of formula (I):

[0007]

[0008] The substituted alcohol nepetalactone compound is a monoterpene compound, which has a substituted hydroxyl group at the 1st position of the six-membered ring structure, a relative molecular weight of 182.09, good water solubility, and a single chemical structure. The substituted hydroxy nepetalactone IUPAC standard name is 4,7-dimethyl-5,6,7,7-tetrahydro-cyclopenta-pyran-1-ol.

[0009] The beneficial effects of the substituted alcohol nepetalactone compound of the present invention are as follows: the present invention uses a zebrafish inflammation model and for the first time finds that the substituted alcohol nepetalactone has excellent anti-inflammatory and analgesic effects, significantly reduces the mRNA levels of TNF-α, NF-κB, and LDHA in the zebrafish inflammation model, and significantly reduces the increase in the movement distance of zebrafish caused by glacial acetic acid pain, and has a significant effect at low concentrations. It has the characteristics of not being easy to produce resistance, being safe, efficient, and low cost, and can become a new alternative drug for anti-inflammatory and analgesic, and is expected to assist in the treatment of inflammation.

[0010] Based on the above technical solution, the present invention can also be improved as follows.

[0011] In a second aspect, a use of a substituted alcohol nepetalactone compound is provided, wherein the substituted alcohol nepetalactone compound described in claim 1 is used to prepare a drug for treating anti-inflammatory and analgesic effects.

[0012] Furthermore, the substituted alcohol nepetalactone compound is used to prepare a drug for treating peripheral nerve pain and inflammatory response.

[0013] In the third aspect, an anti-inflammatory and analgesic drug comprises the substituted alcohol nepetalactone compound. The active ingredient of the effective dose of the anti-inflammatory and analgesic drug is selected from: the substituted alcohol nepetalactone compound of the present invention or its pharmaceutically acceptable salt or pharmaceutically acceptable esterified derivative.

[0014] Furthermore, it also includes pharmaceutically acceptable pharmaceutical excipients and / or carriers.

[0015] Furthermore, the dosage form of the drug is any one of granules, capsules, nanoformulations, controlled-release preparations, and sustained-release preparations.

[0016] In a fourth aspect, a method for preparing a substituted alcohol nepetalactone compound comprises the following steps:

[0017] (1) drying and crushing Nepeta tenuifolia, adding distilled water, introducing process gas to perform steam distillation once, and obtaining a primary extract; introducing process gas into the extract again, performing steam distillation twice, and obtaining a secondary extract; the Nepeta tenuifolia is aboveground Nepeta tenuifolia harvested between September and October; the particle size of the dried and crushed Nepeta tenuifolia is about 10 mesh;

[0018] (2) adding a solvent and silica gel to the secondary extract and then loading the extract onto a column by dry method, degreasing the extract with a degreasing agent, eluting the extract with an eluent, and monitoring the extract with a color developer by TLC to obtain a product solution;

[0019] (3) distilling the product solution (for example, using a rotary evaporator to recover the solvent) to obtain a substituted alcohol nepetalactone compound.

[0020] The beneficial effects of the preparation method of the present invention are: a pure product of substituted alcohol nepetalactone is obtained for the first time through chemical separation operation, and biological experiments show that the extract has good anti-inflammatory and analgesic effects; the extraction method of substituted alcohol nepetalactone provided by the present invention is to obtain the product from the Chinese medicinal herb nepetalactone by steam distillation, primary purification, secondary oxidation, and chromatography purification using a new extraction device, and the obtained product has high yield and high purity.

[0021] It has the characteristics of high efficiency, low cost, high purity and obvious efficacy, and is suitable for large-scale preparation.

[0022] Further, in step (1), the ratio of the amount of Nepeta tenuifolia to the amount of distilled water is 100 g: (100-300) mL; the process gas is oxygen, and the amount of oxygen introduced is 0.05 m 3 / h~0.08m 3 / h.

[0023] Furthermore, in step (2), the ratio of the secondary extract to the solvent is 100 g: (50-150) mL; the solvent includes ethyl acetate;

[0024] The mass ratio of the secondary extract to the silica gel in step (2) is 1:(3-4), and the particle size of the silica gel is 100-200 mesh;

[0025] The degreasing agent in step (2) is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 1:1;

[0026] The eluent in step (2) is a mixture of petroleum ether and ethyl acetate in a volume ratio of 20:1;

[0027] The color developer in step (2) is a sulfuric acid ethanol solution with a mass content of 10%.

[0028] The beneficial effect of adopting the above further scheme is: since the nepeta plant also contains substances such as E, Z-nepetalactone, pseudonepetalactone, citronellol, etc., this brings great difficulties to the separation of the product; the present invention finds through a lot of exploration and research that the substituted alcohol nepetalactone is degreased in the first elution with a solvent ratio of petroleum ether: ethyl acetate at a volume ratio of 1:1, and the second elution is carried out with a solvent ratio of petroleum ether: ethyl acetate at a volume ratio of 20:1; then the nepetalactone crystal product is obtained by recovering the solvent; and a better separation effect is obtained.

[0029] Furthermore, the amount of the degreasing agent in step (2) is 5 to 20 times the column volume; the amount of the eluent is 5 to 20 times the column volume. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1The liquid chromatogram of the product obtained in Example 1 is shown in FIG.

[0031] Figure 2 This is a crystal microscopic image of the product obtained in Example 1.

[0032] Figure 3 This is the concentration-lethal curve of zebrafish in Experimental Example 1.

[0033] Figure 4 This is a typical graph showing the changes in zebrafish neutrophil fluorescence after sample treatment in Experimental Example 1.

[0034] Figure 5 This is a typical graph of the changes in the mRNA levels of inflammatory factors in zebrafish after sample treatment in Experimental Example 1.

[0035] Figure 6 This is a typical diagram of the movement trajectory of zebrafish after sample processing in Experimental Example 1.

[0036] Figure 7 This is a structural diagram of the extraction device of the present invention.

[0037] Among them, 1. electric heating element, 2. first flask, 3. first gas interface, 4. dropping funnel, 5. solid feeding funnel, 6. distillation head, 7. thermometer or thermocouple, 8. condenser, 9. tail pipe, 10. second gas interface, 11. second flask. DETAILED DESCRIPTION

[0038] The principles and features of the present invention are described below, and the examples are only used to explain the present invention and are not used to limit the scope of the present invention. If no specific technology or conditions are specified in the embodiments, the technology or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased through regular channels.

[0039] Example 1

[0040] This embodiment relates to a method for preparing a substituted alcohol nepetalactone compound, comprising the following steps:

[0041] The steam distillation apparatus used is as follows Figure 7As shown, it includes an electric heating element (1), a first flask (2), a first gas interface (3), a dropping funnel (4), a solid feeding funnel (5), a distillation head (6), a thermometer or a thermocouple (7), a condenser (8), a tail pipe (9), a second gas interface (10), and a second flask; the electric heating element (1) receives the first flask (2), a side port of the first flask (2) is connected to the first gas interface (3) and the dropping funnel (4), the solid feeding funnel (5) is connected to the side port of the first flask (2), the middle port of the first flask (2) is connected to the distillation head (6) and the thermometer or the thermocouple (7), the side port of the distillation head (6) is connected to the condenser (8) and the tail pipe (9), the second gas interface (10) is sintered and welded on the tail pipe (9), and receives the second flask to form an airtight distillation and extraction device.

[0042] After the aboveground parts of Nepeta tenuifolia (Nepeta tenuifolia is the aboveground parts of Nepeta tenuifolia harvested between September and October) are dried, crushed and passed through a 10-mesh sieve. 100 g of the above-mentioned sample powder is taken and oxygen is introduced into the first gas interface (3) of the extraction device at a flow rate of 0.05 m / s. 3 / h, steam distillation is performed in a steam distillation device to obtain a primary extract. Take 100g of the primary extract, introduce oxygen into the first gas interface (3) of the extraction device, and set the flow rate to 0.05m 3 / h, add 200ml of distilled water using a dropping funnel (4), connect the condenser (8) to a circulation device, start the electric heating element (1) for secondary steam distillation, and obtain 95g of secondary extract in the second flask (11). Take 100g of the secondary extract, add 100ml of ethyl acetate and 400g of silica gel to mix the sample, dry-load the column, use 10 times the column volume of degreasing agent for degreasing, further use 10 times the column volume of eluent for elution, and use 10% sulfuric acid ethanol for TLC monitoring to obtain a product solution. The particle size of the silica gel is 100-200 mesh; the degreasing agent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 1:1; the eluent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 20:1; take the above product solution, use a rotary evaporator to recover the solvent, and obtain about 5.52g of substituted alcohol nepetalactone monomer. The purity is 97.5% and the yield is 5.38%.

[0043] Example 2

[0044] This embodiment relates to a method for preparing a substituted alcohol nepetalactone compound, comprising the following steps:

[0045] The steam distillation apparatus used is as follows Figure 7The aboveground materials of Nepeta tenuifolia were dried and crushed through a 10-mesh sieve, and oxygen was introduced into the first gas interface (3) of the extraction device at a flow rate of 0.05 m / s. 3 / h, take 100g of the sample powder, and steam distill it in a steam distillation device to obtain a primary extract. Take 100g of the primary extract, and introduce oxygen into the first gas interface (3) of the extraction device at a flow rate of 0.08m 3 / h, add 300ml of distilled water using a dropping funnel (4), connect the condenser (8) to a circulation device, start the electric heating element (1) for secondary steam distillation, and obtain 95g of secondary extract in the second flask (11). Take 100g of the secondary extract, add 100ml of ethyl acetate and 400g of silica gel to mix the sample, dry-load the column, use 10 times the column volume of degreasing agent for degreasing, further use 10 times the column volume of eluent for elution, and use 10% sulfuric acid ethanol for TLC monitoring to obtain a product solution. The particle size of the silica gel is 100-200 mesh; the degreasing agent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 1:1; the eluent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 20:1; take the above product solution, use a rotary evaporator to recover the solvent, and obtain about 6.78g of substituted alcohol nepetalactone monomer. The purity is 96.8% and the yield is 6.56%.

[0046] Example 3

[0047] This embodiment relates to a method for preparing a substituted alcohol nepetalactone compound, comprising the following steps:

[0048] The steam distillation apparatus used is as follows Figure 7 (Same as Example 1). The aboveground materials of Schizonepeta were dried and crushed to pass through a 10-mesh sieve. 100 g of the sample powder was taken and oxygen was introduced into the first gas interface (3) of the extraction device at a flow rate of 0.05 m / s. 3 / h, steam distillation was carried out in a steam distillation device to obtain a primary extract. 100 g of the above extract was taken, and oxygen was introduced into the first gas interface (3) of the extraction device at a flow rate of 0.08 m 3 / h, add 200ml of distilled water using a dropping funnel (4), connect the condenser (8) to a circulation device, start the electric heating element (1) for secondary steam distillation, and obtain 95g of secondary extract in the second flask (11). Take 100g of the secondary extract, add 100ml of ethyl acetate and 300g of silica gel to mix the sample, dry-load the column, use 10 times the column volume of degreasing agent for degreasing, further use 10 times the column volume of eluent for elution, and use 10% sulfuric acid ethanol for TLC monitoring to obtain a product solution. The particle size of the silica gel is 100-200 mesh; the degreasing agent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 1:1; the eluent is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 20:1; take the above product solution, use a rotary evaporator to recover the solvent, and obtain about 1.14g of substituted alcohol nepetalactone monomer. The purity is 95.2% and the yield is 1.08%.

[0049] Test example: Evaluation of the anti-inflammatory and analgesic effects of substituted alcohol nepetalactones

[0050] 1. Experimental Animals

[0051] The experimental transgenic neutrophil fluorescent zebrafish (MPO-eGFP, purchased from Hangzhou Huante Biotechnology Co., Ltd.) were bred by natural pair mating. The age was 10 dpf (i.e., 10 days after fertilization) and used for the anti-inflammatory drug efficacy evaluation experiment.

[0052] AB zebrafish (purchased from Hangzhou Huante Biotechnology Co., Ltd.) were bred by natural pair mating. The age was 3 dpf (3 days after fertilization) and used for molecular biology experiments and peripheral analgesic pharmacological evaluation experiments.

[0053] Culture conditions: Zebrafish were cultured in fish farming water at 28 degrees Celsius (water quality: 200 mg instant sea salt was added to 1 L reverse osmosis water, conductivity was 480-510 uS / cm; pH was 6.9-7.2; hardness was 53.7-71.6 mg / L (CaCO3).

[0054] 2. Experimental Materials

[0055] Test sample: substituted alcohol nepetalactone prepared by the present invention, prepared into 100 mg / L mother solution with DMSO, stored at -15°C. Drugs: copper sulfate (Beijing Chemical Plant Co., Ltd.), dimethyl sulfoxide (DMSO, batch number BCCD9082, Sigma, Switzerland).

[0056] Analytical balance (JJ224BC, Shuangjie Company, USA), pipette (SteoMate Stepper, China), zebrafish aquatic ecosystem (PENTAIR, US), stereo fluorescence microscope SZX7 (Olympus, Japan), zebrafish behavior analysis system (Zebra Lab 3.22.3.31, Viewpoint, France), fluorescence quantitative PCR instrument (Roche, US), CCD camera (Olympus, Japan).

[0057] 3. Experimental methods

[0058] (1) Determination of maximum tolerated concentration (MTC): 3dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). Substituted alcohol nepetalactone was given in water, and a normal control group (WT) and a model control group were set up at the same time. The volume of each well was 3mL. After treatment at 28℃ for 24h, copper sulfate saline solution (5mg / L) was injected into all experimental groups except the normal control group to establish the model. After treatment at 28℃ for another 24h, the MTC of the sample on the model zebrafish was determined. The statistical results were expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0059] (2) Evaluation of the anti-inflammatory effect of substituted alcohol nepetalactone: 10dpf neutrophil fluorescence zebrafish were randomly selected in a 6-well plate, and one zebrafish was treated in each well (experimental group). Substituted alcohol nepetalactone was given in water, and a normal control group (WT) and a model control group were set up at the same time, with a capacity of 3mL per well. After treatment at 28℃ for 24h, copper sulfate saline solution (5mg / L) was injected into all experimental groups except the normal control group to establish the model. After treatment at 28℃ for another 24h, photos were taken under a fluorescence microscope, and the data were processed and recorded. The statistical results were expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0060] (3) Effect of substituted alcohol nepetalactone on the expression of inflammatory factors in zebrafish model: 3dpf neutrophil fluorescence zebrafish were randomly selected in a 6-well plate, and one zebrafish was treated in each well (experimental group). Substituted alcohol nepetalactone was given in water, and a normal control group (WT) and a model control group were set up at the same time, with a capacity of 3mL per well. After treatment at 28℃ for 24h, copper sulfate saline solution (5mg / L) was injected into all experimental groups except the normal control group to establish the model. After treatment at 28℃ for another 24h, qRT-PCR was performed respectively.

[0061] The total RNA was extracted according to the kit product manual, and the RNA content of the extracted product was determined using a super-nano spectrophotometer, where A260 / A280 should be between 2.0 and 2.5. The RNA was further reverse transcribed into cDNA according to the method described in the reverse transcription kit manual. The qRT-PCR operation was performed according to the qRT-PCR instrument instructions, and the primer sequences used are detailed in Table 1. After the amplification reaction was completed, the Ct values ​​of gene C1 and the internal reference gene β-actin in the experimental group, control group, and model group were output, and the relative quantification method 2 -ΔΔCt The relative expression of genes in each group was measured. The data were processed and recorded. The statistical results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis. p < 0.05 indicated that the difference was statistically significant.

[0062] Table 1 Primer sequences used

[0063]

[0064] (4) Evaluation of the analgesic effect of substituted alcohol nepetalactone: 5dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well. Substituted alcohol nepetalactone was given in water, and a normal control group (WT) and a model control group were set up at the same time, with a capacity of 3mL per well. After treatment at 28℃ for 24h, all experimental groups except the normal control group were injected with glacial acetic acid saline solution (5mg / L) to establish a model. After further treatment at 28℃ for 24h, the total movement distance of the zebrafish was measured using a behavioral analyzer, and the statistical analysis results of this indicator were used to evaluate the efficacy of the sample on the inflammatory response induced by copper sulfate. The statistical analysis results were expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0065] 4. Experimental results

[0066] (1) In this experiment, the results are shown in Table 2 and Figure 3 The MTC of substituted alcohol nepetalactone for model zebrafish is 100 mg / L.

[0067] Table 2 Concentration exploration test results (n = 30)

[0068]

[0069]

[0070] (2) As shown in Table 3 and Figure 4, the number of neutrophils in the inflammatory area of ​​zebrafish in the model group was compared with that in the normal control group: P<0.01, indicating that the model was successfully established; the anti-inflammatory efficacy of substituted alcohol nepetalactone 10ug / mL, 50ug / mL and 100ug / mL groups was 42%, 71% and 96%, respectively; compared with the model group, 10ug / mL group P<0.05, 50ug / mL and 100ug / mL groups P<0.01, indicating that substituted alcohol nepetalactone has a more obvious anti-inflammatory activity at low concentrations.

[0071] Table 3 Sample anti-inflammatory effect evaluation test results (n = 30)

[0072]

[0073] (3) As shown in Table 4 and Figure 5 Compared with the control group, the mRNA levels of TNF-α, NF-κB, and LDHA in the model group were significantly upregulated, indicating that the model was successfully established; compared with the model group, the mRNA levels of TNF-α, NF-κB, and LDHA in the 10mg / L, 25mg / L, and 50mg / L substituted alcohol nepetalactone groups were downregulated and statistically significant; the mRNA levels of NF-κB and LDH in the 10mg / L and 25mg / L groups were significantly downregulated. This shows that substituted alcohol nepetalactone can alleviate the abnormal expression of inflammation-related genes induced by copper sulfate.

[0074] Table 4 Experimental results of sample-related inflammatory factor mRNA levels (n=30)

[0075]

[0076] (4) As shown in Table 5 and Figure 6 Compared with the model group, the zebrafish in the positive control group 10mg / L aspirin group had a pain relief rate of about 60%, P<0.01. The pain reduction rates of the 5mg / L, 10mg / L, and 50mg / L groups were about 40%, 105%, and 150%, respectively; compared with the model group, the 5ug / mL, 10ug / mL, and 50ug / mL groups had a P<0.05. The results showed that the substituted alcohol nepetalactone had a more obvious analgesic effect and had a good analgesic effect at a low concentration.

[0077] Table 5 Changes in zebrafish movement length after sample treatment

[0078]

[0079] In summary, the substituted alcohol nepetalactone of the present invention has relatively obvious anti-inflammatory and analgesic activities at low concentrations, and can achieve the anti-inflammatory purpose by inhibiting the mRNA levels of TNF-α, NF-κB, and LDHA at low concentrations.

[0080] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A substituted alcohol nepetalactone compound, characterized in that: The substituted alcohol nepetalactone compound includes a compound as shown in formula (I) or a pharmaceutically acceptable salt or pharmaceutically acceptable esterified derivative of formula (I):

2. An application of a substituted alcohol nepetalactone compound, characterized in that: The substituted alcohol nepetalactone compound according to claim 1 is used for preparing a drug for treating anti-inflammatory and analgesia.

3. The use of a substituted alcohol nepetalactone compound according to claim 2, characterized in that: The substituted alcohol nepetalactone compound is used for preparing medicine for treating peripheral nerve pain and inflammatory reaction.

4. An anti-inflammatory and analgesic drug, characterized in that: The invention comprises the substituted alcohol nepetalactone compound as claimed in claim 1.

5. The anti-inflammatory and analgesic drug according to claim 4, characterized in that: Also included are pharmaceutically acceptable excipients and / or carriers.

6. The anti-inflammatory and analgesic drug according to claim 4, characterized in that: The dosage form of the medicine is any one of granules, capsules, nano preparations, controlled release preparations and sustained release preparations.

7. A method for preparing a substituted alcohol nepetalactone compound according to claim 1, characterized in that: The steps include: (1) drying and crushing Nepeta tenuifolia, adding distilled water, introducing process gas to perform steam distillation once, and obtaining a primary extract; introducing process gas into the extract again, performing steam distillation twice, and obtaining a secondary extract; (2) adding a solvent and silica gel to the secondary extract and then loading the extract onto a column by dry method, degreasing the extract with a degreasing agent, eluting the extract with an eluent, and monitoring the extract with a color developer by TLC to obtain a product solution; (3) distilling the product solution to obtain a substituted alcohol nepetalactone compound.

8. The method for preparing a substituted alcohol nepetalactone compound according to claim 7, characterized in that: In step (1), the ratio of the amount of Nepeta tenuifolia to the amount of distilled water is 100 g: (100-300) mL; the process gas is oxygen, and the amount of oxygen introduced is 0.05 m 3 / h~0.08m 3 / h.

9. The method for preparing a substituted alcohol nepetalactone compound according to claim 7, characterized in that: The ratio of the secondary extract to the solvent in step (2) is 100 g: (50-150) mL; the solvent includes ethyl acetate; The mass ratio of the secondary extract to the silica gel in step (2) is 1:(3-4), and the particle size of the silica gel is 100-200 mesh; The degreasing agent in step (2) is obtained by mixing petroleum ether and ethyl acetate in a volume ratio of 1:1; The eluent in step (2) is a mixture of petroleum ether and ethyl acetate in a volume ratio of 20:1; The color developer in step (2) is a sulfuric acid ethanol solution with a mass content of 10%.

10. The method for preparing a substituted alcohol nepetalactone compound according to claim 7, characterized in that: The amount of the degreasing agent in step (2) is 5 to 20 times the column volume; the amount of the eluent is 5 to 20 times the column volume.

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