CN108101892B - Chrysin non-natural amino acid derivative and preparation method and application thereof - Google Patents

Abstract

The invention discloses chrysin unnatural amino acid derivatives or pharmaceutically acceptable hydrates and salts thereof, including stereoisomers or tautomers thereof. The preparation method comprises the following steps: the chrysin and bromopropyne are subjected to alkali action to prepare 5-hydroxy-7-propargyloxy flavone; (L/D) -halo-Boc amino acid methyl ester with NaN₃Preparing (L/D) -N under the action of CuI/L-sodium prolinate₃-Boc amino acid methyl ester; 5-hydroxy-7-propargyloxyflavone and (L/D) -N₃-Boc amino acid methyl ester and then CuSO₄.5H₂Reacting under the action of O/sodium ascorbate, removing Boc group, and hydrolyzing to obtain chrysin amino acid derivative. The chrysin non-natural amino acid derivative has an anti-cancer effect and can be used for treating anti-cancer drugs.

Description

Chrysin non-natural amino acid derivative and preparation method and application thereof

Technical Field

The invention relates to a chrysin unnatural amino acid derivative and application thereof in pharmacy, belonging to the technical field of medicines.

Background

Chrysin (5, 7-dihydroxyflavone) is a flavone compound widely existing in nature, and has a wide range of pharmacological activities such as antibacterial, antioxidant, antitumor, anti-inflammatory, antidiabetic, anxiolytic and the like. Recent studies have shown that it can also prevent organ toxicity caused by cisplatin and improve cognitive deficits and brain damage caused by intermittent hypoxia. However, because of its poor water solubility, it is poorly absorbed by the intestinal tract and is subject to metabolic inactivation in the body. In order to improve the pharmacological activity, the structure modification and reconstruction are carried out on the compound, and the compound has important significance for obtaining novel high-efficiency low-toxicity candidate drugs.

Amino acids are the basic units constituting proteins, and are important active molecules in the human body and participate in various life activity processes. The amino acid has good solubility and affinity, and the tumor cells have higher requirement on the amino acid than normal cells. The introduction of the compound into drug molecules is beneficial to the effect of the drug reaching cells, thereby improving the selectivity of the drug molecules to tumor cells.

The 1, 2, 3-triazole is an important link unit for drug synthesis, and many compounds containing triazole have various biological activities of resisting fungi, bacteria, tuberculosis, cancer, virus, inflammation and pain, convulsion and the like. The invention combines unnatural amino acid and chrysin molecule through 1, 2, 3-triazole ring, hopes to improve the solubility of chrysin, reduce the killing effect on normal cells and improve the anticancer activity.

Disclosure of Invention

The invention aims to provide a chrysin unnatural amino acid derivative which has an anti-cancer effect.

The invention also aims to provide a preparation method of the chrysin unnatural amino acid derivative.

The invention also aims to provide application of the chrysin unnatural amino acid derivative.

The present invention is described in detail below.

The chrysin non-natural unnatural amino acid derivative or pharmaceutically acceptable hydrate thereof provided by the invention comprises stereoisomers or tautomers thereof, and the structure is shown as follows:

in the formula (I), the compound is shown in the specification,

；

the configuration of the unnatural amino acid is L-type and D-type.

The specific structural examples of the chrysin unnatural amino acid derivatives are as follows:

the invention also provides a preparation method of the compound, which comprises the following steps:

in the formula (I), the compound is shown in the specification,

;

the configuration of the unnatural amino acid is L-type and D-type; z₁Independently of one another Cl, Br, I, Z₂Is H; z₁H, OH, Z₂Each independently is Cl, Br, I; z₃Is N₃When Z is₄Is H; z₃H, OH, Z₄Is N₃(ii) a n is 0, 1.

The chrysin unnatural amino acid derivative or the pharmaceutically acceptable hydrate thereof, including the stereoisomer or the tautomer, has an anti-tumor effect.

The present invention is further illustrated by the following examples, but it should be noted that the scope of the present invention is not limited in any way by these examples.

Detailed Description

Example 1

Preparation of Compound (1)

2.5g (10mmol) of chrysin and 3.36g (20 mmol) of K₂CO₃Adding the mixture into a reactor containing 100mL of anhydrous acetone, slowly adding 2 mL of bromopropyne under vigorous stirring, and carrying out reflux reaction for 3 h. After the reaction, 50mL of water was added, acetone was removed under reduced pressure, the mixture was filtered, and the filter cake was washed with water, 1M HCl solution and water in this order and dried to give a yellow solid as compound (I) in 95% yield. mp is 153.7-154.5 ℃;¹HNMR(CDCl₃,400MHz)δ:12.72(s,1H), 7.89(d,J=4Hz,2H),7.53(m,3H), 6.68(s,1H),6.59(d,J=1.6Hz,1H),6.45(d,J = 2.0 Hz,1H),4.77(d,J = 2.0 Hz,2H),2.61(s, 1H)；ESI-MS( m/z): 293[M+H]⁺。

3.58g (10mmol) of (L) -4-Br-Boc-PheOMe and 0.78g (12 mmol) of NaN₃Adding into 15ml of LDMSO, adding 0.19g (1 mmol) of CuI and 0.41g (3 mmol) of L-proline sodium, heating to 90 deg.C, reacting for 5 hr, adding 50mLH₂O, filtering, and adding 50mLCH₂Cl₂Separating, washing organic layer with saturated saline solution for 3 times, concentrating, and purifying by column chromatography to obtain (L) -4-N₃Boc-PheOMe, yield 86%.

3.84g (12 mmol) of (L) -4-N₃Boc-PheOMe and 2.92g (10mmol) of compound (I) were added to 100mL of a 1: 1 of tert-Butanol and water, and adding, while stirring, 39.6mg (0.2 mmol) of sodium ascorbate and 25mg (0.1 mmol) of CuSO₄·5H₂O, reacting for 5 hours under the condition of vigorous stirring at 50 ℃. The reaction was cooled to room temperature and 100mL of CH was added₂Cl₂Separating with 20mL of 2mol/L HCl solution, washing the organic phase with water for 2 times, drying, filtering, and concentrating under reduced pressure to obtain a crude product. Purifying by column chromatography to obtain intermediate (III). The yield thereof was found to be 85%.

20mL of saturated HCl ethyl acetate solution was added to 5.98g (10mmol) of intermediate (III), the mixture was stirred at room temperature for 5 hours, the mixture was evaporated to dryness under reduced pressure, 20mL of methanol and 20mL of a 2mol/L NaOH solution were added to the residue, the mixture was stirred at room temperature for 2 hours, the pH was adjusted to 3 with dilute hydrochloric acid, the mixture was concentrated under reduced pressure, and the product was purified by column chromatography to obtain compound (1) with a yield of 86%. ESI-MS (M/z):499[ M +1 ]]⁺。Anal. Calcd. for

C₂₇H₂₂N₄O₆: C, 65.05; H, 4.45; N, 11.24; found C, 65.08; H, 4.42; N, 11.26.

Example 2

Preparation of Compound (2)

The same procedures as in example 1 were repeated except for using (D) -4-Br-Boc-PheOMe instead of (L) -4-Br-Boc-PheOMe to obtain compound (2) in 86% yield.

Example 3

Preparation of Compound (3)

The compound (3) was obtained in 88% yield by the same operation as in example 1 except that (L) -4-Br-Boc-PheOMe was replaced with (L) -3-I-Boc-PheOMe.

Example 4

Preparation of Compound (4)

The same procedures as in example 1 were repeated except for using (D) -3-Cl-Boc-phenylglycine methyl ester in place of (L) -4-Br-Boc-PheOMe to obtain compound (4) in 77% yield. ESI-MS (M/z):485[ M +1 ]]⁺。Anal. Calcd. For C₂₆H₂₀N₄O₆: C, 64.46; H, 4.16; N, 11.56; found C, 64.43; H, 4.15; N, 11.56.

Example 5

Preparation of Compound (5)

The same procedures as in example 1 were repeated except for using (L) -3-Cl-Boc-tyrosine methyl ester instead of (L) -4-Br-Boc-PheOMe to obtain compound (5) in 74% yield. ESI-MS (M/z) 515[ M +1 ]]⁺。Anal. Calcd. For C₂₇H₂₂N₄O₇: C, 63.03; H, 4.31; N, 10.89; found C, 63.01; H, 4.34; N, 10.86.

Example 6

Preparation of Compound (6)

The same procedures as in example 1 were repeated except for using (D) -4-Cl-Boc-phenylglycine methyl ester in place of (L) -4-Br-Boc-PheOMe to obtain compound (6) in 79% yield.

Example 7

Inhibition of HepG2 and MGC-803 by chrysin unnatural amino acid derivatives

Culturing human liver cancer cell HepG2 and human stomach cancer cell MGC-803 in 10% volume fraction of fetal calf serum and 1x10⁵U/L penicillin and 1x10⁵PRM1-1640 medium of U/L streptomycin at 37 ℃ with 5% CO₂Cultured in an incubator. Tumor cells in logarithmic growth phase were seeded in 96-well culture plates, and 100. mu.L of single cell suspension (5X 10) was added to each well³Cells), 24h, discarding the supernatant, adding 200. mu.L of drug-containing medium to each well, so that the final concentration of HepG2 cell line compound is 120, 60, 30, 15 and 7.5. mu. mol/L, the final concentration of MGC-803 cell line compound is 20,10,5,2.5 and 1.25. mu. mol/L, cis-platinum is taken as a positive control, the concentration of HepG2 cell line cis-platinum is 100,50,25,12.5 and 6.25. mu. mol/L, the concentration of MGC-803 cell line cis-platinum is 20,10,5,2.5 and 1.25. mu. mol/L, the concentration of the solvent control group is a culture solution containing 0.2% DMSO and 0.1% Tween-80, and the zero adjustment group is a culture solution with only the same amount. Setting 5 multiple wells per concentration, culturing for 48h, adding 20 μ LMTT (5g/L) solution per well, mixing, and culturing for 4 h. Removing culture medium in the wells, adding 150 μ L DSMSO into each well, shaking for 10min to dissolve the blue-violet crystal, measuring OD value at 570nm with microplate reader, and performing with SPSS18.0 softwareStatistical analysis of the data, determination of IC50 value.

The results show that the chrysin unnatural amino acid derivatives have good inhibitory effect on HepG2 cells and MGC-803 cells (Table 1).

Claims (3)

1. The chrysin unnatural amino acid derivative is shown as the following formula:

in the formula (I), the compound is shown in the specification,

the configuration of the unnatural amino acid is L-type and D-type.

2. The chrysin unnatural amino acid derivative according to claim 1, wherein specific examples of the compound include:

。

3. the method for preparing the non-natural amino acid derivative of chrysin according to claim 1, comprising the steps of:

in the formula (I), the compound is shown in the specification,

the configuration of the unnatural amino acid is L-type and D-type; z₁Independently of one another Cl, Br, I, Z₂Is H; z₁H, OH, Z₂Each independently is Cl, Br, I; z₃Is N₃When Z is₄Is H; z₃H, OH, Z₄Is N₃(ii) a n is 0, 1.