CN110818626B - Pyridine carboxylic acid beta-caryophyllene-5-ester compound and preparation method and application thereof - Google Patents
Pyridine carboxylic acid beta-caryophyllene-5-ester compound and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a pyridine carboxylic acid beta-caryophyllene-5-ester compound, a preparation method and application thereof, and belongs to the technical field of preparation of carboxylic acid beta-caryophyllene-5-ester. The preparation method of the pyridine carboxylic acid beta-caryophyllene-5-ester compound comprises the steps of reacting pyridine carboxylic acid with DCC, then adding beta-caryophyllene alcohol and DMAP for reaction, and obtaining the compound after the reaction is finished. NO inhibition rate experiments, cytotoxicity experiments and anticancer activity experiments prove that the compounds have good anti-inflammatory activity and anticancer activity on various cancers, and can be applied to the preparation of anti-inflammatory and anticancer drugs.
Description
Technical Field
The invention belongs to the technical field of preparation of carboxylic acid beta-caryophyllene-5-ester, and particularly relates to a picolinic acid beta-caryophyllene-5-ester compound and a preparation method and application thereof.
Background
Along with the improvement of the quality of life of people, the requirements of people on health are higher and higher, and along with a series of problems caused by the side effect of synthetic drugs, people look to natural product compounds. Beta-caryophyllene is a sesquiterpene compound widely existing in plants. Beta-caryophyllene (also called beta-clove oil) is a bicyclic sesquiterpenoids and widely exists in natural plants such as sweetgum resin, lemon, clove leaf oil, blackcurrant and the like. The pharmacological aspects of the compound have various activities such as local anesthesia, antibiosis and anti-inflammation (Kim, Y.S., et al. journal of Food Science, 2008.73(7), C540-C545. doi: 10.1111/j.1750-3841.2008.00879.x), bone protection and the like (Oh, M. -S., et al. last Management Science, 2013.70(5), 757-762. doi: 10.1002/ps.3608).
For the current research, the research on the beta-caryophyllene mostly focuses on the pharmacology of the beta-caryophyllene. In 2016, Fidyt, K. reported that β -caryophyllene and its oxides have anticancer and analgesic effects (Fidyt, K., et al. cancer Medicine, 2016.5(10), 3007-3017. doi: 10.1002/cam 4.816). In the same year, Basha, R.H. reported that beta-caryophyllene can effectively reduce hyperglycemia-mediated oxidative stress and inflammatory stress in experimental diabetic rats (Basha, R.H. chemical-Biological Interactions, 2016.245, 50-58. doi: 10.1016/J.cbi.2015.12.019). In 2019, Korean researchers Shim, H, found that beta-caryophyllene was effective in inhibiting helicobacter pylori infection, thereby improving symptoms of dyspepsia (Shim, H.et al. the Korean J oumal of Gastroenterology, 2019.74(4), 199. doi: 10.4166/kJ g.2019.74.4.199). Although the research on beta-caryophyllene is more, the research on modification of beta-caryophyllene is less, the structural modification of beta-caryophyllene mainly focuses on epoxidation of double bonds on the ring of beta-caryophyllus, and active oxygen is introduced into beta-caryophyllene through epoxidation so as to improve the effects of the beta-caryophyllene in aspects of anti-inflammation, bacteriostasis, analgesia and the like (Park, k. -R., et al. cancer Letters, 312(2), 178-188. doi: 10.1016/j. canlet.2011.08.001). However, the introduction of pyridine rings into β -caryophyllene by chemical means has not been reported. Only in 2014, Chicca, A synthesized a beta-caryophyllenol carboxylate with a benzene ring by using benzoyl chloride and studied on the inhibition effect of cyclooxygenase, and found that Chicca, A., et al, ACS Chemical Biology, 2014.9 (7), 1499-1507. doi: 10.1021/cb500177c could not effectively inhibit the production of cyclooxygenase. This synthesis requires pyridine as a solvent, which is highly toxic, and acid chlorides are environmentally unstable.
Disclosure of Invention
The invention aims to solve the technical problem of providing the pyridine carboxylic acid beta-caryophyllene-5-ester compound which is a novel compound and has inhibitory activity on inflammation and various tumor cells. The invention aims to solve another technical problem of providing a preparation method of the pyridine carboxylic acid beta-caryophyllene-5-ester compound, which is simple and has high product yield. The invention also provides an application of the picolinic acid beta-caryophyllene-5-ester compound, the compound has good inhibitory activity on inflammation and various cancer cells, and can be applied to preparation of anti-inflammatory and anti-cancer drugs.
The technical scheme is as follows: in order to solve the problems, the technical scheme adopted by the invention is as follows:
the pyridine carboxylic acid beta-caryophyllene-5-ester compound has a structural formula shown in a formula I:
wherein the R group is:
the preparation method of the pyridine carboxylic acid beta-caryophyllene-5-ester compound comprises the following steps:
(1) dissolving picolinic acid and DCC in CH2Cl2Performing reaction at 0-5 ℃ for 20-30 min; the molar ratio of DCC to picolinic acid is 1: 1-1: 2, and the concentration of DCC is 0.15-0.20 mol/L;
(2) dissolving DMAP in CH 2Cl2Adding the DMAP solution and beta-caryophyllenol into the solution obtained in the step (1), and reacting for 4-5 h at 25-35 ℃; the molar ratio of the DMAP to the beta-caryophyllenol is 1: 15-1: 20, and the concentration of the DMAP is 0.02-0.08 mol/L; the molar ratio of the picolinic acid to the beta-caryophyllenol is 1-3: 1;
(3) and (3) washing the reaction solution after the reaction section is finished, drying, removing the solvent, and eluting by using a silica gel column to obtain the picolinic acid beta-caryophyllene-5-ester compound.
The preparation method of the beta-caryophyllene-5-picolinate compound comprises the following steps:
(a) dissolving the secondary naphthol borane in tetrahydrofuran to obtain a secondary naphthol borane solution, adding beta-caryophyllene, and carrying out reflux reaction at 70-90 ℃ for 17-19 h; the molar ratio of the naphthol borane to the beta-caryophyllene is 1-2: 1, and the concentration of the naphthol borane solution is 1-2 mol/L;
(b) after the reaction is finished, CH is adopted2Cl2Diluting and cooling, and sequentially adding KOH solution and 30% H2O2Reacting for 20-40 min; the molar ratio of the KOH to the beta-caryophyllene is 6.5-7.0: 1, and the concentration of the KOH solution is 3-4 mol/L; the KOH solution was mixed with 30% H2O2The volume ratio of (A) to (B) is 1: 1-1: 2;
(c) and after the reaction is finished, washing the product for 3 times by using saturated NaCl, drying the product, removing the solvent to obtain dark yellow oily liquid, eluting the dark yellow oily liquid by using a 100-200-mesh silica gel column chromatography, and obtaining light yellow oily liquid beta-caryophyllenol by using a mobile phase petroleum ether and ethyl acetate which are 1: 7.
The picolinic acid beta-caryophyllene-5-ester compound is prepared by using pyridine-2-formic acid, pyridine-3-formic acid or pyridine-4-formic acid.
The preparation method of the pyridine carboxylic acid beta-caryophyllene-5-ester compound comprises the following steps of (1) reacting at 0 ℃ for 30 min; and (2) reacting at 25 ℃ for 5 h.
According to the preparation method of the picolinic acid beta-caryophyllene-5-ester compound, the molar ratio of the picolinic acid to the beta-caryophyllenol is 1: 1.
According to the preparation method of the pyridine carboxylic acid beta-caryophyllene-5-ester compound, the molar ratio of DCC to pyridine carboxylic acid is 1: 1, and the concentration of DCC is 0.18 mol/L; the molar ratio of the DMAP to the beta-caryophyllenol is 1: 18, and the concentration of the DMAP is 0.05 mol/L.
The picolinic acid beta-caryophyllene-5-ester compound is applied to preparing anti-inflammatory drugs.
The picolinic acid beta-caryophyllene-5-ester compound is applied to the preparation of anti-cancer drugs.
The picolinic acid beta-caryophyllene-5-ester compound is applied to preparation of anti-cancer drugs, wherein the cancers are cervical cancer, liver cancer, breast cancer or lung cancer.
Has the advantages that: compared with the prior art, the invention has the advantages that:
(1) the preparation method of the picolinic acid beta-caryophyllene-5-ester compound has simple reaction steps and mild reaction conditions.
(2) The picolinic acid beta-caryophyllene-5-ester compound prepared by the invention can effectively inhibit the generation of NO of inflammatory cells, has small damage to the cells and has good application prospect in the preparation of anti-inflammatory drugs.
(3) The beta-caryophyllene-5-picolinate compound prepared by the invention has good inhibitory activity on cervical cancer, liver cancer, breast cancer and lung cancer cells, and has good application prospect in preparation of anti-cancer drugs.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.
Example 1
Synthesis method of beta-caryophyllenol C1(6, 10, 10-trimethyl-2-methylidenebic [7.2.0] uncacan-5-ol):
adding 4.4mmol of beta-caryophyllene into 6.6mL of 1M tetrahydrofuran solution of naphthol borane, carrying out reflux reaction at 80 ℃ for 18h, and reacting with 40mL of CH2Cl2Diluting and cooling, adding 20mL of 3M KOH and 20mL of 30% H in sequence2O2And reacting for 30min, washing with saturated NaCl for 3 times, drying, removing the solvent to obtain dark yellow oily liquid, eluting with a 100-200-mesh silica gel column, and obtaining light yellow oily liquid C1 by using a mobile phase of petroleum ether and ethyl acetate at a ratio of 1: 7. The reaction equation is as follows:
C1 1HNMR(600M，DMSO-d6)δ：4.83(d，2H，＝CH2，J＝6Hz)，4.22(d，1H，-OH，J＝6Hz)，3.32(s，1H，-CH)，2.46-2.42(m，1H，-CH)，2.22-2.12(m，2H，-CH2)，1.92-1.86(m，1H，-CH)，1.74-1.71(m，2H，-CH2)，1.70(t，1H，J＝6Hz，-CH)，1.58-1.53(m，1H，-CH)，1.51-1.48(m，2H，-CH2)，1.44-1.39(m，2H，-CH2)，0.38(s，3H，-CH3)，0.97(s，3H，-CH3)，0.96-0.95(m，1H，-CH)，0.83(d，3H，-CH3，J＝6Hz)；13CNMR(DMSO-d6，150MHz)：153.05，108.59，56.83，55.20，42.02，36.75，35.64，33.87，32.6，30.78，30.25，26.84，21.87；IR，υ(cm-1)：3372(-OH)，3076(C＝CH2)997(C-O)；Elemental Anal.Calcd for C15H26O：C 81.02；H 11.79；O 7.19；Found：C 81.14；H 11.82；O7.21.HRMS(ESI+)：m/z[C15H26O+H]+223.2056 test value 223.2064.
Synthesis method of pyridine-2-carboxylic acid beta-caryophyllene-5-ester C2(6, 10, 10-trimethyl-2-methylinebicyclo [7.2.0] undecan-5-ylpicolinate):
pyridine-2-carboxylic acid 0.9mmol and dicyclohexylcarbodiimide (DCC, 0.9mmol) were dissolved in 5mL of CH2Cl2Reacting at 0 ℃ for 30 min; c10.9mmol was added and dissolved in 1mL CH2Cl24-dimethylamino pyridine (DMAP, 0.05mmol), reacting for 5h at normal temperature, washing, drying, removing the solvent, and passing through a silica gel column to obtain yellow oily liquid C2130 mg. The reaction equation is as follows:
C2 1HNMR(600M，CDCl3)δ：8.71(d，1H，-CH，J＝4.2)，8.01(d，1H，J＝7.8Hz)，7.78-7.77(m，1H，-CH)，7.40-7.38(m，1H，-CH)，4.88-4.80(m，2H，＝CH2)，3.52-3.49(m，1H，-CH)，2.43-2.37(m，1H，-CH)，2.23-2.14(m，2H，-CH2)，1.81-1.78(m，1H，-CH)，1.73-1.68(m，2H，-CH2)，1.60-1.55(m，2H，-CH2)，1.55-1.52(m，2H，CH2)，1.473-1.45(m，1H，-CH)，0.97(d，3H，CH3，J＝3.6Hz)，0.94(s，3H，-CH3)，0.916(s，1H，-CH)，0.90-0.87(m，3H，CH3)；13CNMR(CDCl3，150MHz)δppm：164.51，152.67，150.01，136.86，126.57，124，89，108.60，56.58，56.50，42.10，36.78，35.4135.14，33.92，33.83，31.93，29.99，26.80，21.48，21.43.IR，υ(cm-1)：3076(C＝CH2)，1714(C＝O)，1460(C＝N，C-C)，303(C-O)，1134(C-O-C)；Elemental Anal.Calcd for C21H29NO2：C 77.02；H 8.93；N 4.28；O9.77；Found：C 77.13；H 9.02；N 4.37；O 9.84.HRMS(ESI+)：m/z[C21H29NO2+H+]+328.2271，[C21H29NO2+Na+]350.2091 test value 328.2280, 350.2099.
Example 2
Synthesis method of 3-picolinic acid beta-caryophyllene-5-ester C3(6, 10, 10-trimethyl-2-methylinebicyclo [7.2.0] undecan-5-yl nicotinate):
pyridine-3-carboxylic acid 0.9mmol and DCC 0.9mmol were dissolved in 5mL of CH2Cl2Reacting at 0 ℃ for 30 min; c10.9mmol prepared in example 1 and dissolved in 1mL CH2Cl20.05mmol of DMAP, reacting for 5 hours at normal temperature, washing, drying and removing the solvent. After passing through a silica gel column, 140mg of yellow oily liquid was obtained. The reaction equation is as follows:
C3 1HNMR(600M，CDCl3)δ：9.23(d，1H，CH，J＝1.2Hz)，8.77-8.76(m，1H，CH)，8.30-8.28(m，1H，CH)，7.40-7.38(m，1H，CH)，4.93-4.84(m，2H，＝CH2)，2.35-2.33(m，1H，CH)，2.25-2.18(m，1H，CH)，2.09-2.05(m，1H，CH)，1.93-1.87(m，1H，CH)，1.81-1.77(m，2H，CH2)，1.76-1.73(m，2H，CH2)1.53-1.50(m，1H，CH)，1.25(s，1H，CH)，1.03(d，3H，CH3，J＝1.2Hz)，0.99(s，3H，CH3)，0.97(t，1H，CH，J＝0.6Hz)，0.932-0.913(m，3H，CH3)；13CNMR(CDCl3，150MHz)δppm：164.73，153.04，151.98150.70，137.18，126.85，123.36，109.39，56.52，42.19，36.82，35.03，33.94，31.45，29.99，28.58，26.81，21.45.IR，υ(cm-1)：3076(C＝CH2)，1718(C＝O)，1460(C＝N，C-C)，1134(C-O-C)；Elemental Anal.Calcd for C21H29NO2：C 77.02；H 8.93；N 4.28；O 9.77；Found：C 77.15；H 9.06；N 4.32；O 9.86.HRMS(ESI+)：m/z[C21H29NO2+H+]+328.2271，[C21H29NO2+Na+]350.20916，[C21H29NO2+K+]366.1830 test value 328.2279, 350.2099, 366.2047.
Example 3
Synthesis method of 4-picolinic acid beta-caryophyllene-5-ester C4(6, 10, 10-trimethyl-2-methylinebicyclo [7.2.0] undecan-5-ylisonicotinate):
Pyridine-4-carboxylic acid 0.9mmol and DCC 0.9mmol were dissolved in 5mL of CH2Cl2And reacting at 0 ℃ for 30 min. C10.9mmol prepared in example 1 and dissolved in 1mL CH2Cl20.05mmol of DMAP, reacting for 5 hours at normal temperature, washing, drying and removing the solvent. After passing through a silica gel column, 136mg of yellow oily liquid was obtained. The reaction equation is as follows:
C4 1HNMR(600M，CDCl3)δ：8.76(d，2H，CH，J＝5.4Hz)，7.84(d，2H，CH，J＝6Hz)，4.928-4.843(m，2H，＝CH2)，2.46-2.42(m，1H，CH)，2.34-2.32(m，1H，CH)，2.22-2.16(m，1H，CH)，2.07-2.00(m，1H，CH)，1.91-1.86(m，1H，CH)，1.81-1.75(m，2H，CH2)，1.73-1.75(m，1H，CH)，1.64-1.62(m，2H，CH2)，1.52-1.50(m，1H，CH)，1.24(s，1H，CH)，1.02(d，3H，CH3，J＝1.8Hz，)，0.99(s，3H，CH3)，0.96(t，1H，J＝1.8Hz，CH)，0.915(t，3H，CH3，J＝7.2Hz).13CNMR(CDCl3，150MHz)δppm：164.62，152.71，151.92，150.51，138.17，122.89，109.47，108.61，56.60，42.12，36.79，35.03，33.85，31.97，31.42，29.99，28.53，26.82，21.46.IR，υ(cm-1)：3072(C＝CH2)，1724(C＝O)，1460(C＝N，C-C)，1124(C-O-C)；Elemental Anal.Calcd for C21H29NO2：C 77.02；H 8.93；N 4.28；O 9.77；Found：C 77.12；H 8.99；N 4.22；O 9.76.HRMS(ESI+)：m/z[C21H29NO2+H+]+328.2271，[C21H29NO2+K+]366.1830 test value 328.2285, 366.2052.
Example 4
First, experiments on NO inhibition rates of compounds C2-C4:
(1) taking a mouse macrophage RAW264.7 with logarithmic growth cycle, inoculating 3-4 ten thousand of the macrophage RAW in each hole in a 96-hole plate, and inoculating 5% CO at 37 DEG C2Incubating for 24 hours in an incubator; removing the culture medium after incubation is finished, and washing 3-4 times by using PBS;
(2) setting a control group, an LPS + Dexamethasone (DIM) positive drug group and a compound C2-C4 (the concentration is 40,20, 10, 5, 2.5 mu M) sample group; experimental groups are shown below:
control group: 1. adding 50 μ L of 2 μ g/mL LPS and 50 μ L of caryophyllene C0 with concentration of 40,20, 10, 5, 2.5 μ M respectively into each well; 2. adding 50 μ L of 2 μ g/mL LPS and 50 μ L of beta-caryophyllenol C1 with concentration of 40,20, 10, 5, 2.5 μ M respectively into each well;
LPS + DIM positive drug group: add 50. mu.L of LPS 2. mu.g/mL and 50. mu.L DIM at concentrations of 40,20, 10, 5, 2.5. mu.M, respectively, to each well;
Compound C2 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C2 per well;
compound C3 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C3 per well;
compound C4 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C4 per well;
adding the liquid of the experimental groups in sequence, and continuously incubating in the incubator for 24 h;
(3) centrifuging to take cell culture supernatant to a 96-hole mildew standard plate, and sequentially adding A and B of the ELISA kit according to the volume ratio of 1: 1; reacting for 3min in dark place, and testing absorbance at 540nm with a mold standard instrument; the formula for calculating the NO inhibition rate is shown in the following formula (1):
the results of the NO inhibition test for compounds C2-C4 are shown in Table 1. It can be seen from table 1 that the modified compounds C1, C2, C3 and C4 have a better effect on the inhibition of NO. Compared with unmodified C0 (beta-caryophyllene), C2, C3 and C4 have better anti-inflammatory activity. And the inhibition effect of C3 and C4 on NO is about 1.2-1.3 times that of positive control DIM. This shows that the anti-inflammatory activity of C0 can be effectively improved by introducing pyridine heterocycle on beta-caryophyllene.
TABLE 1 NO inhibition (%) test results for Compounds C2-C4
Second, cytotoxicity test for RAW264.7 for compounds C2 to C4:
(1) taking RAW264.7 of logarithmic growth cycle, inoculating 3-4 ten thousand of RAW in each hole in a 96-hole plate, and inoculating 5% CO at 37 DEG C2After incubation for 24h in an incubator, removing the culture medium, and washing with PBS 3-4 times;
(2) setting a control group, an LPS + DIM positive drug group and a compound C2-C4 sample group; experimental groups are shown below:
control group: 1. adding 50 μ L of 2 μ g/mL LPS and 50 μ L of caryophyllene C0 with concentration of 40, 20, 10, 5, 2.5 μ M respectively into each well; 2. adding 50 μ L of 2 μ g/mL LPS and 50 μ L of beta-caryophyllenol C1 with concentration of 40, 20, 10, 5, 2.5 μ M respectively into each well;
LPS + DIM positive drug group: add 50. mu.L of LPS 2. mu.g/mL and 50. mu.L DIM at concentrations of 40, 20, 10, 5, 2.5. mu.M, respectively, to each well;
compound C2 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C2 per well;
compound C3 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C3 per well;
compound C4 sample set: add 50. mu.L of 2. mu.g/mL LPS and 50. mu.L of 40, 20, 10, 5, 2.5. mu.M C4 per well;
Continuously incubating in the incubator for 24h, removing the supernatant, adding 1mg/mL MTT dye, and incubating for 4 h;
(3) after the incubation is finished, removing the culture solution, adding 200 mu L DMSO, shaking the plate at 37 ℃ for 10min, detecting the absorbance at 595 position by using a mildew standard instrument, and calculating the cell survival rate according to the formula (2):
the results of the cell viability assay for compounds C2-C4 are shown in table 2. As can be seen from table 2, it was found that β -caryophyllene (C2, C3, and C4) introduced with a pyridine ring can effectively reduce the cytotoxicity of β -caryophyllene by cytotoxicity tests of C0 to C4 and a positive control DIM. And has better cell survival rate compared with a positive control. This shows that the introduction of pyridine ring can effectively reduce the cytotoxicity of natural products, thereby providing a certain data support for the patent drug of beta-caryophyllene.
TABLE 2 cytotoxicity test results (% cell viability) of Compounds C2-C4
Thirdly, testing the anticancer activity of the compounds C2-C4 by an MTT method:
(1) taking Hela (cervical cancer cell), HepG2 (liver cancer cell), MCF-7 (breast cancer cell), A549 (lung cancer cell) and HUVEC (human umbilical vein endothelial cell) with logarithmic growth cycle, inoculating the cells in a 96-well plate in 1-2 ten thousand per well, and inoculating 5% CO at 37 ℃ in 5% 2The incubator is incubated for 24 hours;
(2) removing the culture medium, adding diluted samples of a control group and an experimental group, incubating for 48 hours, removing the culture medium, adding 1mg/mL MTT, and incubating for 4 hours; wherein, the control group adopts DOX (doxycycline), and the concentration of the samples of the control group and the experimental group is 100, 10, 1, 0.1 and 0.01 mu M;
(3) after the incubation is finished, removing the culture medium, adding 200 mu L DMSO, shaking the plate at 37 ℃ for 10min, detecting the absorbance at 595 position by a mildew standard instrument, and calculating the cell inhibition rate according to the formula (3):
the results of the antitumor activity tests of compounds C2-C4 are shown in Table 3. In Table 3, the anti-tumor activity tests of the compounds C0-C4 show that C1, C2 and C3 have characteristic inhibition effects on MCF-7 and have small toxic and side effects on HUVEC. IC thereof50Both values were lower than C0 and the positive control DOX. This shows that the killing effect of beta-caryophyllene on cancer cells can be effectively improved by introducing pyridine rings, and the method has a high safety factor.
TABLE 3 half inhibitory concentration IC of Compounds C2-C4 on cancer cells50μM
Claims (10)
1. The pyridine carboxylic acid beta-caryophyllene-5-ester compound is characterized in that the structural formula is shown as a formula I:
wherein the R group is:
2. the preparation method of the beta-caryophyllene-5-picolinate ester compound as claimed in claim 1, comprising the steps of:
(1) Dissolving picolinic acid and DCC in CH2Cl2Performing reaction at 0-5 ℃ for 20-30 min; the molar ratio of DCC to picolinic acid is 1: 1-1: 2, and the concentration of DCC is 0.15-0.20 mol/L;
(2) dissolving DMAP in CH2Cl2Adding the DMAP solution and beta-caryophyllenol into the solution obtained in the step (1), and reacting for 4-5 hours at the temperature of 25-35 ℃; the molar ratio of the DMAP to the beta-caryophyllenol is 1: 15-1: 20, and the concentration of the DMAP solution is 0.02-0.08 mol/L; the molar ratio of the picolinic acid to the beta-caryophyllenol is 1-3: 1;
(3) and after the reaction is finished, washing the reaction solution, drying, removing the solvent, and eluting by using a silica gel column to obtain the picolinic acid beta-caryophyllene-5-ester compound.
3. The method for preparing the beta-caryophyllene-5-pyridinecarboxylate compound according to claim 2, characterized in that the preparation of the beta-caryophyllene alcohol comprises the following steps:
(a) dissolving the secondary naphthol borane in tetrahydrofuran to obtain a secondary naphthol borane solution, adding beta-caryophyllene, and carrying out reflux reaction at 70-90 ℃ for 17-19 h; the molar ratio of the naphthol borane to the beta-caryophyllene is 1-2: 1, and the concentration of the naphthol borane solution is 1-2 mol/L;
(b) after the reaction is finished, CH is adopted 2Cl2Diluting and cooling, and sequentially adding KOH solution and 30% H2O2Reacting for 20-40 min; the molar ratio of the KOH to the beta-caryophyllene is 6.5-7.0: 1, and the concentration of the KOH solution is 3-4 mol/L; what is neededKOH solution with 30% H2O2The volume ratio of (A) to (B) is 1: 1-1: 2;
(c) and after the reaction is finished, washing the mixture for 3 times by using saturated NaCl, drying the mixture, removing the solvent to obtain dark yellow oily liquid, eluting the dark yellow oily liquid by using a 100-200-mesh silica gel column, and obtaining light yellow oily liquid beta-caryophyllenol by using a mobile phase petroleum ether and ethyl acetate which are 1: 7.
4. The method for preparing the beta-caryophyllene-5-picolinate compound according to claim 2 or 3, wherein the picolinic acid is pyridine-2-carboxylic acid, pyridine-3-carboxylic acid or pyridine-4-carboxylic acid.
5. The method for preparing the beta-caryophyllene-5-pyridinecarboxylate compound according to claim 2 or 3, characterized in that in the step (1), the reaction is carried out at 0 ℃ for 30 min; and (2) reacting for 5h at 25 ℃.
6. The method for preparing the beta-caryophyllene-5-picolinate compound according to claim 2 or 3, wherein the molar ratio of the picolinic acid to the beta-caryophyllene alcohol is 1: 1.
7. The method for preparing the beta-caryophyllene-5-picolinate compound according to claim 2 or 3, wherein the molar ratio of DCC to picolinic acid is 1: 1, and the concentration of DCC is 0.18 mol/L; the molar ratio of the DMAP to the beta-caryophyllenol is 1: 18, and the concentration of the DMAP is 0.05 mol/L.
8. The use of beta-caryophyllene-5-pyridinecarboxylate compounds according to claim 1 for the preparation of anti-inflammatory drugs.
9. The use of the beta-caryophyllene picolinate-5-ester compound of claim 1 in the preparation of anti-cancer drugs.
10. The use of the beta-caryophyllene-5-pyridinecarboxylate compound according to claim 9 in the preparation of an anti-cancer drug, wherein the cancer is cervical cancer, liver cancer, breast cancer, or lung cancer.
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Non-Patent Citations (5)
| Title |
|---|
| Afifa Zardi-Bergaoui等.Caryophyllene Sesquiterpenes from Pulicaria vulgaris GAERTN.:Isolation, Structure Determination, Bioactivity and Structure Activity Relationship.《Chemistry & Biodiversity》.2019,第16卷(第2期), * |
| Functionalization of β‑Caryophyllene Generates Novel Polypharmacology in the Endocannabinoid System;Andrea Chicca等;《ACS Chemical Biology》;20140505;第9卷(第7期);第1499-1507页 * |
| NO inhibitory constituents as potential anti-neuroinflammatory agents for AD from Blumea balsamifera;Jun Ma等;《Bioorganic Chemistry》;20171205;第76卷;第449-457页 * |
| β-石竹烯生物学功能的研究进展;张季林，等;《山东医药》;20181231;第58卷(第38期);第110-112页 * |
| β-石竹烯的研究进展;陈旭冰，等;《山东化工》;20111231;第40卷;第34-36页 * |
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