Application of tryptophan derivatives of parthenolide in the preparation of glioma drugs

Abstract

The present invention discloses an application of a tryptophan derivative of parthenolide in the preparation of a glioma drug, and belongs to the field of pharmaceutical technology. In view of the problems of poor water solubility and metabolic instability of existing parthenolide and its derivatives, the present invention proposes a tryptophan derivative PDTry-TZ with significant anti-tumor activity. The derivative has an IC50 value as low as 3.352µM / ml, can significantly inhibit the growth of tumor cells, and is prepared by a specific synthesis method. The tryptophan derivative PDTry-TZ of parthenolide provides a new drug candidate for the treatment of gliomas, which is expected to improve the therapeutic effect, reduce side effects, and improve the quality of life of patients. In summary, the present invention not only solves the problems of the prior art, but also brings new breakthroughs in the field of glioma treatment, and has important application value.

Description

Technical Field

[0001] The invention belongs to the field of medicines, and in particular relates to the application of a tryptophan derivative of parthenolide in the preparation of glioma medicines. Background Art

[0002] Tumors pose a great threat to human health. According to relevant statistics, 7.6 million people die from cancer every year worldwide.

[0003] Glioma is a general term for tumors that originate from glial cells and neuronal cells in the nervous system. It is the most common malignant tumor in the brain, accounting for 40%-50% of intracranial tumors. Glioma can be divided into astrocytoma, glioblastoma, oligodendroglioma and other types according to the cell type. The treatment and prognosis of different types of glioma are also different.

[0004] Parthenolide (PTL) is a sesquiterpenoid compound extracted from Asteraceae plants. It has unique anti-inflammatory and anti-tumor biological activities, and can significantly inhibit the growth of cancer cells such as glioma, thymic carcinoma, colorectal cancer, and lymphoma, and promote their apoptosis. PTL has a unique chemical structure and can inhibit the expression of cell transcription factors NF-κB, STATs, and JNK, and induce ROS response, which may be the molecular mechanism of PTL's anti-inflammatory and anti-tumor functions. PTL is non-toxic to normal cells, but shows a broad spectrum of cytotoxicity and pro-apoptotic effects on a variety of tumor cells, suggesting its development prospects as an important tumor suppressor drug.

[0005] However, the currently disclosed parthenolide and its derivatives have problems such as poor water solubility, unstable metabolism, excessive dosage, and low bioavailability. Summary of the invention

[0006] The present invention provides an application of a tryptophan derivative of parthenolide in the preparation of a glioma drug, so as to solve the problems of the existing parthenolide and its derivatives, such as poor water solubility, unstable metabolism, excessive dosage, low bioavailability, etc.

[0007] The present invention provides application of a tryptophan derivative of parthenolide in preparing a glioma drug.

[0008] The purpose of the present invention is to provide an application of a tryptophan derivative of parthenolide in the preparation of glioma drugs.

[0009] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0010] IC of the present invention 50 It is 3.352, indicating that the tryptophan derivatives of parthenolide can significantly inhibit the growth of tumor cells.

[0011] The present invention provides an application of a tryptophan derivative of parthenolide in the preparation of a glioma drug. By synthesizing a new tryptophan derivative PDTry-TZ, tumor cell growth is significantly inhibited and anti-tumor activity is improved, thereby providing a new drug candidate for the treatment of glioma. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a synthetic route of the tryptophan derivative of parthenolide provided in an embodiment of the present invention;

[0013] Figure 2 The IC of the tryptophan derivative PDTry-TZ of parthenolide provided in the embodiment of the present invention is 50 Line chart;

[0014] Figure 3 1H NMR of product PDTry-TZ (400 MHz, CDCl3) provided in an embodiment of the present invention;

[0015] Figure 4 The embodiment of the present invention provides 13 Schematic representation of C NMR of product PDTry-TZ (100 MHz, CDCl3). DETAILED DESCRIPTION

[0016] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings. It should be understood that these descriptions are only exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, the description of well-known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0017] Synthesis steps of tryptophan derivative PDTry-TZ of parthenolide:

[0018] 1. Specific steps for the synthesis of intermediate 6:

[0019]

[0020] .

[0021] At room temperature, dry dichloromethane was used as solvent, selenium dioxide and tert-butyl peroxide were combined to carry out regioselective oxidation. 14The allylic methyl group of the product was oxidized to produce a hydroxyl group, and a mixed derivative of hydroxyl group and aldehyde was obtained. The reaction mixture was diluted with 50 ml of dichloromethane, and 100 ml of saturated sodium thiosulfate aqueous solution was added to the reaction solution. The organic phase was separated by storage and extraction, and the extraction was repeated 3 times. After the organic phases were combined, they were filtered through diatomaceous earth, and the filtrate was concentrated by rotary evaporation under reduced pressure; it was purified by column chromatography on silica gel, and eluted with ethyl acetate: dichloromethane (1:10) to obtain C 14 The oxidation product (Melampomagnolide B) MMB, i.e. compound 2.

[0022] Compound 2 (1.14 mmol, 300.00 mg, 1.00 eq) was dissolved in 7 ml DMF, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.26 mol, 1.11 eq) and 1-hydroxybenzotriazole (1.26 mmol, 1.1 eq) were added, and after stirring for 5 min, Boc-tryptophan (1.3 eq) was added, and N,N-diisopropylethylamine (3.05 eq) was slowly added dropwise while stirring. The reaction system was placed at room temperature and reacted overnight. The reaction mixture was diluted with 50 ml of dichloromethane, neutralized and extracted twice with saturated NaHCO3 aqueous solution, the organic phases were combined, and then extracted three times with saturated NaCl aqueous solution, the organic phases were combined again, and dried with anhydrous Na2SO4. Filtered, and the organic solvent was evaporated under reduced pressure using a rotary evaporator. The concentrated mixture was purified by column chromatography on silica gel, eluted with ethyl acetate: petroleum ether (1:4), and the corresponding intermediate 5 was obtained after purification, and weighed for the next reaction. Among them, intermediate 5 was also used as a new compound after derivatization, and the corresponding spectrum collection work was also carried out.

[0023] Under 0℃ ice bath condition, the intermediate 5 was dissolved in dichloromethane. After the reaction solution was cooled to 0℃, trifluoroacetic acid (TFA: DCM = 1: 1) was added dropwise to the reaction solution. After stirring for 2h, the reaction solution was neutralized with a saturated NaHCO3 aqueous solution until no bubbles were generated, and the reaction was terminated. The reaction solution was diluted with 50ml of dichloromethane and extracted with purified water for 3 times. The organic phases were combined and dried with anhydrous Na2SO4, filtered, and the organic solvent was removed by rotary evaporation under reduced pressure. The concentrated mixture was purified by column chromatography on silica gel, ethyl acetate: dichloromethane (1: 8) to obtain the target compound, i.e., the tryptophan derivative of parthenolide.

[0024] 2. The synthetic route of the tryptophan derivative PDTry-TZ of parthenolide is as follows Figure 1 shown.

[0025] Weigh the intermediate 6 (0.0463mmol, 19.00mg, 1.1eq), 3-(3-(but-3-yn-1-yl)-3H-naphthyridin-3-yl)propionic acid (0.0421mmol, 7.00mg, 1eq), HoBt (0.1053mmol, 14.00mg, 2.4eq), EDCI (0.0632mmol, 12mg, 1.5eq) in turn, place in a dry sealed tube, add 2ml DMF to dissolve it, stir for 5 minutes, gradually add DIPEA (0.1306mmol, 17mg, 3.1eq), react for 24h and then detect. The reaction solution was diluted with 10ml dichloromethane, and then extracted with saturated NaHCO3 aqueous solution and saturated NaCl aqueous solution respectively. After extraction, the organic phases were combined and dried with anhydrous Na2SO4. Filter and remove the organic solvent under reduced pressure using a rotary evaporator. The concentrated mixture was separated and purified using a preparative plate, EA:PE (1:1), and finally the tryptophan derivative PDTry-TZ of parthenolide was obtained. The target product was a white powder, such as Figure 1 shown.

[0026] like Figure 3 and Figure 4 As shown, the structure and NMR spectrum data of the tryptophan derivative PDTry-TZ of parthenolide:

[0027]

[0028] The product was obtained in 75.5% yield. 1 H NMR (400 MHz, DMSO) 10.98 (s, 1H), 8.52 (d, J = 6.8 Hz, 1H), 7.51 (d, J = 7.6 Hz, 1H), 7.38 (d, J= 7.9 Hz, 1H), 7.22 (s, 1H), 7.12 (t, J = 7.2 Hz, 1H), 7.04 (t, J = 7.2 Hz, 1H), 6.08 (d, J = 2.3 Hz, 1H), 5.58 (s, 1H), 5.45 (s, 1H), 4.53 (dd, J =13.9, 6.8 Hz, 1H), 4.45 (d, J = 12.3 Hz, 1H), 4.36 (d, J = 12.3 Hz, 1H), 4.09(t, J = 9.1 Hz, 1H), 3.21 – 3.05 (m, 2H), 2.87 (d, J = 24.4 Hz, 2H), 2.68 (d,J = 9.4 Hz, 1H), 2.34 – 2.16 (m, 3H), 2.14 – 1.94 (m,8H), 1.73 – 1.42 (m,8H). 13 C NMR (101 MHz, DMSO) 8.22, 31.36, 29.10,28.25, 28.08, 27.27, 24.69, 23.92, 23.25, 17.47, 172.03, 171.00, 169.52, 139.54, 136.16, 134.54,129.84, 127.06, 123.71, 121.05, 119.44, 118.52, 117.96, 111.50, 109.26,83.21, 80.60, 71.83, 67.46, 62.62, 59.94, 53.51, 41.71, 36.22, 31.36, 29.10,28.25, 28.08, 27.27, 12.71.Chemical Formula:C 34 H 38 N4O6, [M+H] + , 598.279.

[0029] The molecular weights of the compounds were determined by reversed phase high performance liquid chromatography (RP-HPLC) and mass spectrometry (MS).

[0030] Target study of tryptophan derivative PDTry-TZ of parthenolide based on chemical proteomics:

[0031] Experimental research objects, reagents and instruments:

[0032] Human astrocytic glioblastoma cells U87mg were cryopreserved by the School of Pharmacy and Pharmaceutical Sciences, Shandong First Medical University.

[0033] Preparation of stock solution: Accurately weigh 2 mg of the tryptophan derivative PDTry-TZ of parthenolide, add DMSO solution to dissolve it, and prepare a 40 mM stock solution. The stock solution is divided and frozen in a -80℃ refrigerator. Before the experiment, it is diluted with culture medium to the required concentration according to the experimental design.

[0034] Evaluation of the cytotoxic effect of the tryptophan derivative PDTry-TZ molecule of parthenolide:

[0035] Take U87 cells in the logarithmic growth phase, count the cells after digestion, and dilute the cell suspension to a concentration of 1×10 5 / ml, inoculated into 96-well plates, with a volume of 100µL added to each well, and the medium was replaced after 12h. 80µL of fresh complete medium was added to the wells, and the pre-prepared drug storage solution was diluted according to the required concentration, diluted to 200, 100, 50, and 25µmol / L, respectively, and 20µL was added to each well, so that the final concentration was 40, 20, 10, and 5µmol / L per well. Five replicate wells were set for each concentration, and a control group containing only cells and DMSO without drugs was set up; a complete culture medium without cells was set up as a blank group. After culturing in a 37°C, 5% CO2 incubator for 24h, 10µL of CCK-8 solution was added to each well, incubated in the incubator for 2h, and the absorbance (OD) value at 450nm was measured with an enzyme reader, and the survival rate of various cell lines was calculated according to the following formula, and the corresponding IC 50 The experiment was repeated 3 times.

[0036] Cell viability = [(OD of experimental well - OD of blank well) / (OD of control well - OD of blank well)] × 100%.

[0037] like Figure 2 As shown, the experimental results of evaluating the cytotoxic effect of the tryptophan derivative PDTry-TZ molecule of parthenolide:

[0038] Table 1 IC of the tryptophan derivative PDTry-TZ of parthenolide 50 value

[0039]

[0040] Table 2 IC of tryptophan derivatives of parthenolide 50 Value Summary Table

[0041]

[0042] The synthesized compounds were respectively applied to U87, U251, and A172 cell lines for activity spectrum screening. The present invention selected PTL and ACT001, which is currently in phase III clinical trials, as controls. Each cell line IC 50 The values ​​were obtained by repeating the experiment for more than 3 times. From the data in Table 2, it can be seen that the tryptophan derivative of parthenolide, PDTry-TZ, has very good antitumor activity.

[0043] The embodiments described above are merely descriptions of preferred implementations of the present invention and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of a tryptophan derivative of parthenolide in the preparation of a glioma drug, wherein the structural formula of the tryptophan derivative of parthenolide, PDTry-TZ, is: 。

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