CN110872305B

CN110872305B - Fluorocamptothecin medicament derivative and preparation and application thereof

Info

Publication number: CN110872305B
Application number: CN201911158256.4A
Authority: CN
Inventors: 朱勍; 刘江; 窦言东
Original assignee: Zhejiang University of Technology ZJUT
Current assignee: Zhejiang University of Technology ZJUT
Priority date: 2019-11-22
Filing date: 2019-11-22
Publication date: 2021-05-04
Anticipated expiration: 2039-11-22
Also published as: CN110872305A

Abstract

The invention relates to a fluorinated camptothecin drug derivative and preparation and application thereof. The compound is prepared by a direct fluorination method of organic photocatalytic phenol, has simple operation process, mild reaction conditions, high site selectivity and high reaction efficiency, can be prepared by only one step, has better antitumor activity, and provides a new scheme for antitumor drugs.

Description

Fluorocamptothecin medicament derivative and preparation and application thereof

(I) technical field

The invention relates to a fluorinated camptothecin derivative containing drugs, a preparation method thereof and application thereof in preparing antitumor drugs.

(II) background of the invention

Phenol is an important basic organic raw material, and many downstream products thereof are involved in numerous fields, mainly for the manufacture of phenol-formaldehyde resins, bisphenol a and caprolactam. And phenol derivatives such as halophenol, nitrophenol and alkylphenol can be used for the production of medicines, pesticides, paints, dyes, explosives, perfumes and the like. Phenol compounds are widely found in nature, and fragrances and many natural products contain phenol structures. The traditional method for modifying the phenolic compound comprises the following steps: firstly protecting hydroxyl, then modifying other sites of the hydroxyl, and finally removing protection to realize modification. The method has the disadvantages of harsh conditions, more steps and low reaction yield, and the existing problems greatly reduce the progress of the modification research of the phenol compounds.

Disclosure of the invention

The invention aims to provide a fluorinated camptothecin medicament obtained by directly fluorinating ortho positions of phenol compounds, a preparation method thereof and application thereof in preparing antitumor medicaments.

The technical scheme adopted by the invention is as follows:

a fluorinated camptothecin drug derivative has the following structure:

the fluorinated camptothecin drug derivative can be prepared by the following method: reacting the camptothecin drug derivative shown as the formula (II) with a fluorination reagent in a solvent under the action of a photocatalyst and a light source at room temperature, and separating and purifying a reaction mixture after the reaction is completed to obtain the fluorinated camptothecin drug derivative shown as the formula (I);

the fluorinating agent is one of the following: n-fluorobisbenzenesulfonamide (NFSI), 1-chloromethyl-4-fluoro-1, 4-diazotized bicyclic 2.2.2 octane bis (tetrafluoroborate) salt (Selectfluor), 1-fluoropyridinium tetrafluoroborate (N-fluoropyrdiinium);

the solvent is one of the following: acetonitrile, water, acetic acid aqueous solution, dichloromethane, dimethyl sulfoxide and 1, 2-dichloroethane;

the photocatalyst is one of the following: triterpyridyl ruthenium chloride hexahydrate ([ Ru (bpy))₃]Cl₂) Tris (2-phenylpyridine) iridium ([ Ru (bpy))₃]Cl₂) Fluorescein (Fluorescein), Rhodamine b (Rhodamine b), Rhodamine (Rhodamine 6G), eosin y (eosin y);

the light source is one of the following: blue, green, yellow;

the ratio of the phenol compound, the fluorinating agent and the photocatalyst is 1: 1-3: 0.05-0.1.

Preferably, the fluorinating agent is 1-chloromethyl-4-fluoro-1, 4-diazotized bicyclo 2.2.2 octane bis (tetrafluoroborate) salt, the solvent is 10% acetic acid aqueous solution, the photocatalyst is eosin Y, and the substance amount ratio of the camptothecin drug derivative, the fluorinating agent and the photocatalyst is 1:1.5: 0.05.

The separation and purification method comprises the following steps: adding saturated NaCl water solution into the reaction mixture, extracting with ethyl acetate, drying the organic layer with anhydrous sodium sulfate, filtering, and rotary evaporating at normal temperature to remove solvent to obtain crude product; and (3) performing silica gel column chromatography on the crude product, taking a solution with the volume ratio of ethyl acetate to petroleum ether being 1:10 as a mobile phase, tracking and collecting an eluent with the Rf value of 0.3-0.5 by TLC, decompressing and removing the solvent from the collected eluent, and drying to obtain the fluorinated phenol compound shown in the formula (2A).

The antitumor activity of the obtained fluorinated camptothecin drug derivative (I) is subjected to an MTT (methyl thiazolyl tetrazolium) experiment of the antitumor activity of HeLa cells. The experimental result shows that (I) has stronger antitumor activity than the original antitumor drug SN-38.

The invention also relates to application of the fluorinated camptothecin derivative containing drugs in preparation of antitumor drugs.

In particular, the tumor is cervical cancer.

The invention has the following beneficial effects: the compound is prepared by a direct fluorination method of organic photocatalytic phenol, has simple operation process, mild reaction conditions, high site selectivity and high reaction efficiency, can be prepared by only one step, has better antitumor activity, and provides a new scheme for antitumor drugs.

(IV) description of the drawings

FIG. 1 is a nuclear magnetic hydrogen spectrum of compound (I);

FIG. 2 is a carbon spectrum of compound (I);

FIG. 3 is a fluorine spectrum of compound (I);

FIG. 4 is a comparison of the antitumor activities of Compound (I) and the antitumor drug SN-38.

(V) detailed description of the preferred embodiments

The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:

example 1: preparation of Compound (I)

Adding 1mmol of camptothecin derivative SN-38 into 4ml of 10% (W/W) acetic acid aqueous solution, adding 1.5mmol of Selectfluor and 0.05mmol of Eosin Y, irradiating and reacting for 6 hours at room temperature by 12W blue light, adding saturated NaCl aqueous solution into reaction liquid after the reaction is finished, extracting by using dichloromethane, taking an organic layer, drying by anhydrous sodium sulfate, filtering, decompressing and evaporating to dryness, and obtaining a crude compound. And (3) performing silica gel column chromatography on the crude compound, wherein the volume ratio of ethyl acetate to petroleum ether is 1: the solution of 9 is a mobile phase, an eluent with Rf value of 0.3-0.5 is collected by TLC tracking, the collected eluent is decompressed to remove the solvent, and is dried to obtain 252mg (yield 70%) of a pure compound shown as a formula 2A-2, and the nuclear magnetic hydrogen spectrum, the carbon spectrum and the fluorine spectrum of the pure compound are shown in figures 1-3.

¹H NMR(500MHz,DMSO)δ10.29(s,1H),8.01(d,J＝9.0Hz,1H),7.39(d,J＝2.6Hz,1H),6.49(s,1H),5.47–5.35(m,2H),5.23(s,2H),3.06(q,J＝7.5Hz,2H),1.87(ddt,J＝21.5,14.1,7.2Hz,2H),1.30(t,J＝7.6Hz,3H),0.89(t,J＝7.3Hz,3H).¹³C NMR(126MHz,DMSO)δ172.53(s),156.78(d,J＝16.0Hz),150.06(s),148.82(s),146.43(s),143.63(s),142.70(s),131.53(s),128.16(s),127.94(s),122.35(s),117.99(s),104.76(s),95.78(s),72.40(s),65.26(s),49.41(s),30.28(s),22.27(s),13.32(s),7.75(s).¹⁹F NMR(376MHz,DMSO)δ-141.81(s).HRMS(ESI⁺):Calculated for C₂₂H₁₉FN₂O₅:[M+H]⁺411.1278，found 411.1275.

Example 2: detection of antitumor Activity of Compound (I)

Tumor cell Hela (cervical cancer cell) is selected, and the MTT method is adopted to detect the proliferation activity of the anti-tumor cell. The cells were seeded at 4000-5000 cells/well in a 96-well plate containing a 1640 culture medium of 10% fetal bovine serum, and the plate cover was annotated with 5% CO₂Culturing at 37 ℃ for 12 hours, allowing cells to adhere to the wall of a 96-well plate, adding a drug to be tested (the compound (I) prepared in example 1) to a sterile operating platform by using a pipette gun so that the drug concentration in each well is respectively set to five concentration gradients of 0.01. mu.M, 0.1. mu.M, 1. mu.M, 10. mu.M and 100. mu.M, each concentration is provided with five parallel groups, and taking the anti-tumor drug SN-38 as a control), and placing the 96-well plate in a 5% CO solution again₂And incubated at 37 ℃ for 24 hours. The 96-well plate was removed, and 10. mu.L of MTT kit reagent (purchased from Promega) was added to each well, protected from light at 5% CO₂Incubating for 4 hours at 37 ℃, absorbing supernatant, adding 150uL sterile DMSO to dissolve formazan, further dissolving in an incubator at 37 ℃ for 5-10min, and finally measuring the absorbance by using an enzyme-linked immunosorbent assay (ELISA) instrument. Thus, the cytostatic and cytotoxic rates were calculated, processed with GraphPad Prism software, and the IC calculated₅₀And IC₅₀The 95% confidence interval, see figure 4 and table 1.

TABLE 1

Compound (I)	HeLa cell IC₅₀Value (μm)
		Compound (I)	1.562±0.4
SN-38	9.086±0.5

The experimental result shows that the compound (I) has stronger antitumor activity than the original antitumor drug SN-38.

Claims

1. A process for the preparation of a fluorinated camptothecin drug derivative, the process comprising: reacting the camptothecin drug derivative shown as the formula (II) with a fluorination reagent in a solvent under the action of a photocatalyst and a light source at room temperature, and separating and purifying a reaction mixture after the reaction is completed to obtain the fluorinated camptothecin drug derivative shown as the formula (I);

the fluorinating reagent is 1-chloromethyl-4-fluoro-1, 4-diazotized bicyclic 2.2.2 octane bis (tetrafluoroborate) salt;

the solvent is 10% acetic acid water solution;

the photocatalyst is eosin Y;

the light source is one of the following: blue, green, yellow;

the ratio of the camptothecin drug derivative to the fluorination reagent to the photocatalyst is 1: 1-3: 0.05-0.1.

2. The method of claim 1, wherein: the quantity ratio of the camptothecin drug derivative to the fluorination reagent to the photocatalyst is 1:1.5: 0.05.

3. The method according to claim 1, wherein the separation and purification method comprises: adding saturated NaCl water solution into the reaction mixture, extracting with ethyl acetate, drying the organic layer with anhydrous sodium sulfate, filtering, and rotary evaporating at normal temperature to remove solvent to obtain crude product; and (3) performing silica gel column chromatography on the crude product, taking a solution with the volume ratio of ethyl acetate to petroleum ether being 1:10 as a mobile phase, tracking and collecting an eluent with the Rf value of 0.3-0.5 by TLC, decompressing and removing the solvent from the collected eluent, and drying to obtain the camptothecin fluoride drug derivative shown in the formula (I).