CN113683596A

CN113683596A - Pyrimidine tankyrase 2 inhibitor and preparation method and application thereof

Info

Publication number: CN113683596A
Application number: CN202110943449.1A
Authority: CN
Inventors: 庞婉; 李堂; 刘运力; 王杰; 赵泽圣; 张孝礼
Original assignee: Shanghai Institute of Technology
Current assignee: Shanghai Institute of Technology
Priority date: 2021-08-17
Filing date: 2021-08-17
Publication date: 2021-11-23
Anticipated expiration: 2041-08-17
Also published as: CN113683596B

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a pyrimidine tankyrase 2 inhibitor, a preparation method and an application thereof, wherein the inhibitor has the following structure:

compared with the prior art, the preparation method is simple, and the synthesized compound has a novel structure and better activity of inhibiting tankyrase 2.

Description

Pyrimidine tankyrase 2 inhibitor and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a pyrimidine tankyrase 2 inhibitor as well as a preparation method and application thereof.

Background

Tankyrase is a multifunctional protein translation modifying enzyme, a member of the polyadenylic diphosphoribose polymerase family, and its main domain is ankyrin repeat. The enzyme can be combined with a telomere repetitive sequence factor, and can also be combined with a highly conserved structural domain of a carboxyl terminal of an axial inhibitory protein, so that the inhibition effect on the telomere enzyme is realized, and an extracellular factor (Wnt)/beta-catenin (beta-catenin) signal path is adjusted, so that the proliferation and the growth of tumor cells are inhibited. A tankyrase inhibitor XAV939 reported in Nature in 2009 is directly synthesized based on the crystal structure of tankyrase 1/2, and can selectively block the activation of a Wnt/beta-catenin signaling pathway through the expression of a stable axis inhibiting protein, so as to limit the proliferation and growth of tumor cells. The tankyrase 2 inhibitors known at present can be classified into inhibitors acting on the active site of nicotinamide, inhibitors acting on the adenosine binding site, and inhibitors acting on the binding sites of nicotinamide and adenosine, depending on the active site. However, the semi-Inhibitory Concentration (IC) of the tankyrase 2 inhibitor GK-007 has been reported₅₀) Larger, i.e., less sensitive, and therefore the development of novel tankyrase 2 inhibitors is needed.

Disclosure of Invention

The invention aims to provide a pyrimidine tankyrase 2 inhibitor with better activity, a preparation method and application thereof.

With the continuous development of computer technology, virtual screening technology has played an increasingly important role in the field of new drug discovery. In the invention, 350000 small molecule compounds in a ZINC database are screened layer by comprehensively utilizing molecular docking and molecular dynamics simulation technologies to finally obtain a compound with the terminal anchor polymerase 2 inhibition activity theoretically, then the compound is subjected to group modification to design two more ideal compound structures and a synthetic route, and finally the terminal anchor polymerase 2 inhibition activity is tested and compared with the reported inhibition activity (IC) of a terminal anchor polymerase 2 inhibitor GK-007₅₀0.035uM) for comparison.

The purpose of the invention is realized by the following technical scheme:

the invention provides a pyrimidine tankyrase 2 inhibitor, which has a structure shown in a formula I:

wherein R is selected from

Any one of them.

Preferably, R is selected from:

the second aspect of the present invention provides a preparation method of the pyrimidine tankyrase 2 inhibitor, comprising the following steps:

(1): reacting 2,4, 6-trichloropyrimidine with morpholine in an organic solvent under the condition of adding alkali to obtain 4- (2, 6-dichloropyrimidin-4-yl) morpholine;

(2): reacting 4- (2, 6-dichloropyrimidin-4-yl) morpholine with 2-hydroxyethyl pyridine in an organic solvent under the condition of adding alkali to obtain 4- (6-chloro-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine;

(3): reacting 4- (6-chloro-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine with hydrazine hydrate in an organic solvent to obtain an intermediate 4- (6-hydrazino-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine;

(4): 4- (6-hydrazino-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine and an acid corresponding to the R group are subjected to condensation reaction in an organic solvent under the conditions that a uronium salt is used as a condensing agent and a base is added to obtain the pyrimidine telotromonase 2 inhibitor.

In the preparation method of the pyrimidine tankyrase 2 inhibitor, the reaction equation is as follows:

preferably, in the step (1), the molar ratio of the 2,4, 6-trichloropyrimidine to morpholine is 1: 1-2: 1.

preferably, in the step (1), the base is any one selected from sodium hydroxide, potassium carbonate, triethylamine, sodium carbonate and sodium bicarbonate.

Preferably, in the step (1), the organic solvent is selected from any one of methanol, ethanol, DCM and THF.

Preferably, in the step (1), the reaction temperature of the reaction is-20 ℃ to 30 ℃.

Preferably, in the step (2), the molar ratio of the 4- (2, 6-dichloropyrimidin-4-yl) morpholine to the 2-hydroxyethyl pyridine is 1: 1-2: 1.

preferably, in the step (2), the base is selected from any one of sodium hydroxide, potassium hydroxide, sodium hydride and sodium tert-butoxide.

Preferably, in the step (2), the organic solvent is selected from any one of DMF and THF.

Preferably, in the step (2), the reaction temperature of the reaction is-20 ℃ to 30 ℃.

Preferably, in the step (3), the molar ratio of the 4- (6-chloro-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine to the hydrazine hydrate is 1: 1-1: 3.

preferably, in the step (3), the organic solvent is 1, 4-dioxane.

Preferably, in the step (3), the reaction temperature of the reaction is 70-130 ℃.

Preferably, in step (4), the molar ratio of 4- (6-hydrazino-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine to the corresponding acid in group R is 1: 1-2: 1.

preferably, in the step (4), the base is any one of DIPEA, DIEA, TEA and NMM;

preferably, in the step (4), the organic solvent is any one selected from DMF, DCM, 1,4-dioxane and THF.

Preferably, in the step (4), the condensing agent is any one selected from HATU, HBTU, HCTU, TBTU, TSTU and TNTU.

Preferably, in the step (4), the reaction temperature of the reaction is 0-50 ℃.

Preferably, in step (4), the acid corresponding to the R group has the formula:

wherein R is₁Is selected from

Any one of the above;

that is, the formula of the acid corresponding to the R group is:

any one of them.

Further preferably, R₁Is selected from

The third aspect of the invention provides an application of the pyrimidine tankyrase 2 inhibitor in preparation of a drug for inhibiting tankyrase 2.

Compared with the prior art, the invention has the following beneficial effects:

(1) the pyrimidine tankyrase 2 inhibitor synthesized by the invention has better effect, two of the pyrimidine tankyrase 2 inhibitors are optimizedIC of compound₅₀Value (to determine IC)₅₀0.027uM and 0.018uM, respectively) compared to the positive control, GK-007 (IC)₅₀0.035uM), higher sensitivity and better bioactivity.

(2) The compound synthesized by the invention has a novel structure, and is reported for the first time.

(3) The preparation method is simple.

Drawings

FIG. 1 shows nuclear magnetic resonance of a compound of formula IV¹H NMR chart (CDCl)₃)；

FIG. 2 shows NMR of a compound of formula IV¹³C NMR chart (CDCl)₃)；

FIG. 3 shows NMR of a compound of formula V¹H NMR chart (CDCl)₃)；

FIG. 4 shows NMR of a compound of formula V¹³C NMR chart (CDCl)₃)；

FIG. 5 shows NMR of the objective product obtained in example 1¹H NMR graph (DMSO);

FIG. 6 shows NMR of the objective product obtained in example 1¹³C NMR charts (DMSO);

FIG. 7 is an HRMS chart ([ M + Na ] s) of the target product obtained in example 1]⁺(CH₃OH))；

FIG. 8 shows NMR of target product obtained in example 2¹H NMR graph (DMSO);

FIG. 9 shows NMR of target product obtained in example 2¹³C NMR charts (DMSO);

FIG. 10 is the HRMS pattern ([ M + Na ] for the target product obtained in example 2]⁺(CH₃OH))。

Detailed Description

A pyrimidine tankyrase 2 inhibitor having the structure shown in formula I:

wherein R is selected from

Any one of them.

In some embodiments, R is further preferably selected from:

the preparation method of the pyrimidine tankyrase 2 inhibitor comprises the following steps:

The reaction equation is as follows:

in some embodiments, the molar ratio of 2,4, 6-trichloropyrimidine to morpholine in step (1) is 1: 1-2: 1.

in some embodiments, the base in step (1) is selected from any one of sodium hydroxide, potassium carbonate, triethylamine, sodium carbonate and sodium bicarbonate.

In some embodiments, the organic solvent in step (1) is selected from any one of methanol, ethanol, DCM, and THF.

In some embodiments, the reaction temperature of the reaction in step (1) is from-20 ℃ to 30 ℃.

In some embodiments, the molar ratio of 4- (2, 6-dichloropyrimidin-4-yl) morpholine to 2-hydroxyethylpyridine in step (2) is 1: 1-2: 1.

in some embodiments, the base in step (2) is selected from any one of sodium hydroxide, potassium hydroxide, sodium hydride, and sodium tert-butoxide.

In some embodiments, the organic solvent in step (2) is selected from any one of DMF and THF.

In some embodiments, the reaction temperature of the reaction in step (2) is from-20 ℃ to 30 ℃.

In some embodiments, the molar ratio of 4- (6-chloro-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine to hydrazine hydrate in step (3) is 1: 1-1: 3.

in some embodiments, the organic solvent in step (3) is 1, 4-dioxane.

In some embodiments, the reaction temperature of the reaction in step (3) is from 70 ℃ to 130 ℃.

In some embodiments, the molar ratio of 4- (6-hydrazino-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine to the corresponding acid of the R group in step (4) is 1: 1-2: 1.

in some embodiments, the base in step (4) is any one of DIPEA, DIEA, TEA and NMM;

in some embodiments, the organic solvent in step (4) is selected from any one of DMF, DCM, 1,4-dioxane and THF.

In some embodiments, the condensing agent in step (4) is selected from any one of HATU, HBTU, HCTU, TBTU, TSTU and TNTU.

In some embodiments, the reaction temperature of the reaction in step (4) is from 0 ℃ to 50 ℃.

In some embodiments, the acid corresponding to the R group in step (4) has the formula:

wherein R is₁Is selected from

Any one of the above;

that is, the formula of the acid corresponding to the R group is:

any one of them.

In some embodiments, R₁Further preferably selected from:

the pyrimidine tankyrase 2 inhibitor is applied to preparation of a drug for inhibiting tankyrase 2.

The invention is described in detail below with reference to the figures and specific embodiments.

Preparation of a target product:

step (1)

Dissolving 2,4, 6-trichloropyrimidine (4.5g,25mmol) in 25ml ethanol, adding morpholine (1.9g, 22mmol) and triethylamine (5g, 50mmol) to the ethanol solution containing 2,4, 6-trichloropyrimidine under ice bath condition while stirring, slowly heating to room temperature, cooling to room temperatureStirring was continued for 5h until morpholine reaction was complete. After completion of the reaction, the mixture was extracted with ethyl acetate, water and 10% Na₂CO₃And washing with brine. Purification by flash silica gel chromatography (petroleum ether/ethyl acetate 4:1) afforded the product. To obtain the product of formula III.

Step (2)

4- (2, 6-dichloropyrimidin-4-yl) morpholine (3.5g,15mmol) was dissolved in 10ml THF, and 2-hydroxyethylpyridine (1.5g, 12mmol) and sodium hydride (0.6g, 30mmol) were added to a solution of THF containing 4- (2, 6-dichloropyrimidin-4-yl) morpholine under ice-bath conditions with stirring, and stirring continued for 2h under ice-bath conditions until the 2-hydroxyethylpyridine reaction was complete. After the reaction was completed, the reaction solution was poured into ice water, a white solid precipitated, and the solid was washed with water and 10% Na₂CO₃And washing with brine. To obtain the product of formula IV.

Step (3)

4- (6-chloro-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine (2.5g,7.8mmol) was dissolved in 10ml1,4-Dioxane (1, 4-Dioxane) and N was added with stirring₂H₄·H₂O (750mg, 15mmol) was added to a solution of 4- (2, 6-dichloropyrimidin-4-yl) morpholine in 1,4-Dioxane, slowly warmed to 100 ℃ and stirring continued at 100 ℃ for 5h until the reaction of 4- (6-chloro-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine was complete. After the reaction was complete, 1,4-Dioxane (1, 4-Dioxane) was spin dried under vacuum at 40 ℃ and 20mL ethyl acetate was added to the residue, a white solid precipitated which was water and 10% Na₂CO₃And washing with brine. To obtain the product of formula V.

Step (4)

The acid corresponding to the R group (0.5mmol) was dissolved in 10ml DMF and HATU (230mg, 0.6mmol) and DIPEA (130mg, 1mmol) were added to a solution of the acid corresponding to the R group in DMF with stirring under ice bath, slowly warmed to room temperature and after 15 minutes 4- (6-hydrazino-2- (2- (pyridin-2-yl) ethoxy) pyrimidin-4-yl) morpholine (200mg, 0.6mmol) was added. Stirring was continued at room temperature for 6h until the corresponding acid reaction was complete. After completion of the reaction, the reaction solution was poured into ice water. Separating out solid, collecting solid, adding water and 10% Na₂CO₃Saline and ethanol ultrasonic washing. Purification of the solid by flash silica gel chromatography (petroleum ether/ethyl acetate 2:1) afforded the product. To obtain the product of formula IV.

Testing of inhibitory activity of pyrimidine tankyrase:

the inhibitory activity of the compounds on TNKS2 was determined by an enzymatic reaction. Dissolving a compound to be detected in a DMSO solution to prepare a solution with an initial concentration of 200 mu M/L, diluting the solution 5 times each time according to a proportion, and configuring 7 concentration gradients to obtain a solution concentration range of each compound to be detected of 0.0128 mu M/L to 200 mu M/L. The enzymatic reaction was carried out in 96-well plates (Greiner bio-one U-shaped blackboard) at room temperature. TNKS2 at 5nM was incubated with compound and 500nM NAD' in assay buffer (50mM HEPES pH 7.0, 1mM CHAPS) for 1 hour, with four parallel experimental groups being set up simultaneously per concentration gradient. Then, assay buffer (50mM HEPES,0.8M KF and 20mM EDTA in 0.1% BSA) was added to react at room temperature for 1 hour. And finally, adding 20mL of 20% acetophenone ethanol solution and 20mL of 2M KOH to terminate the reaction, chemically converting the unreacted NAD' into a fluorescent substance, and reading the fluorescence intensity under the conditions of an excitation wavelength of 355nm and an absorption wavelength of 450 nm. According to the formula: inhibition rate (sample fluorescence intensity-blank)/(enzyme value fluorescence intensity-blank) the corresponding inhibition rate for each inhibitor was calculated and then half the effective inhibitory concentration was fitted using Graph Pad Prism software.

The structural formulas of formula IV and formula V are as follows:

¹H NMR(501MHz,CDCl₃)δ8.47(d,J＝4.9Hz,1H),7.53(td,J＝7.8,1.9Hz,1H),7.20(d,J＝7.7Hz,1H),7.06(dd,J＝7.4,4.9Hz,1H),6.08(s,1H),4.61(t,J＝6.8Hz,2H),3.67(t,J＝4.9Hz,4H),3.58–3.48(m,4H),3.19(t,J＝6.8Hz,2H).¹³C NMR(126MHz,CDCl₃)δ164.25,164.10,161.09,158.22,149.32,136.38,123.72,121.53,95.28,66.67,66.31,44.45,37.59.

¹H NMR(501MHz,CDCl₃)δ8.52(d,J＝4.4Hz,1H),7.57(td,J＝7.6,1.9Hz,1H),7.23(d,J＝7.8Hz,1H),7.10(dd,J＝7.5,4.9Hz,1H),6.39(s,1H),5.51(s,1H),4.61(t,J＝6.9Hz,2H),3.72(t,J＝4.9Hz,4H),3.57–3.48(m,4H),3.23(t,J＝6.9Hz,2H).¹³C NMR(126MHz,CDCl₃)δ168.29,164.77,164.43,158.80,149.35,136.38,123.64,121.45,75.14,66.59,65.74,44.66,37.90.

the following examples were prepared and tested for inhibitory activity according to the above preparation methods and inhibitory activity test methods.

Example 1

¹H NMR(400MHz,DMSO)δ10.11(s,1H),8.62(s,1H),8.49(d,J＝4.8Hz,1H),7.70(t,J＝7.7Hz,1H),7.31(d,J＝7.7Hz,1H),7.25–7.16(m,3H),6.93(d,J＝8.7Hz,1H),5.26(s,1H),4.69(s,2H),4.50(t,J＝6.6Hz,2H),3.62(t,J＝4.8Hz,4H),3.38–3.28(m,4H),3.11(t,J＝6.8Hz,2H),2.21(s,3H).¹³C NMR(101MHz,DMSO)δ167.97,167.11,164.83,164.46,158.89,155.33,149.59,136.96,130.58,129.31,126.84,124.91,123.93,122.10,113.25,76.46,66.96,66.29,65.44,44.62,37.53,16.42.

IC₅₀The value was 0.027 uM.

Example 2

¹H NMR(400MHz,DMSO)δ10.44(s,1H),8.70(s,1H),8.48(d,J＝4.9Hz,1H),7.68(t,J＝7.6Hz,1H),7.28(d,J＝10.1Hz,3H),7.23–7.19(m,1H),5.44(s,1H),4.50(t,J＝6.9Hz,2H),3.85(s,3H),3.82(s,6H),3.60(t,J＝4.8Hz,4H),3.40–3.36(m,4H),3.10(t,J＝6.8Hz,2H).¹³C NMR(101MHz,DMSO)δ166.13,165.89,164.92,164.59,158.92,153.29,149.62,140.98,136.99,128.16,123.99,122.16,105.52,76.34,66.39,65.54,60.68,56.57,44.73,37.60.

IC₅₀The value was 0.018 uM.

The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims

1. A pyrimidine tankyrase 2 inhibitor having the structure shown in formula i:

wherein R is selected from

Any one of them.

2. The pyrimidine tankyrase 2 inhibitor of claim 1, wherein R is selected from the group consisting of:

3. a method for preparing the pyrimidine tankyrase 2 inhibitor of claim 1 or 2, comprising the steps of:

4. The method for preparing the pyrimidine tankyrase 2 inhibitor according to claim 3, wherein the step (1) comprises any one or more of the following conditions:

(i) the mol ratio of the 2,4, 6-trichloropyrimidine to the morpholine is 1: 1-2: 1;

(ii) the alkali is selected from any one of sodium hydroxide, potassium carbonate, triethylamine, sodium carbonate and sodium bicarbonate;

(iii) the organic solvent is selected from any one of methanol, ethanol, DCM and THF;

(iv) the reaction temperature is-20 ℃ to 30 ℃.

5. The method for preparing the pyrimidine tankyrase 2 inhibitor according to claim 3, wherein the step (2) comprises any one or more of the following conditions:

(i) the mol ratio of the 4- (2, 6-dichloropyrimidin-4-yl) morpholine to the 2-hydroxyethyl pyridine is 1: 1-2: 1;

(ii) the alkali is selected from any one of sodium hydroxide, potassium hydroxide, sodium hydride and sodium tert-butoxide;

(iii) the organic solvent is selected from any one of DMF and THF;

(iv) the reaction temperature is-20 ℃ to 30 ℃.

6. The method for preparing the pyrimidine tankyrase 2 inhibitor according to claim 3, wherein the step (3) comprises any one or more of the following conditions:

(i) the molar ratio of the 4- (6-chloro-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine to the hydrazine hydrate is 1: 1-1: 3;

(ii) the organic solvent is 1, 4-dioxane;

(iii) the reaction temperature of the reaction is 70-130 ℃.

7. The method for preparing the pyrimidine tankyrase 2 inhibitor according to claim 3, wherein the step (4) comprises any one or more of the following conditions:

(i) the molar ratio of the 4- (6-hydrazino-2- (2- (pyridine-2-yl) ethoxy) pyrimidine-4-yl) morpholine to the corresponding acid of the R group is 1: 1-2: 1;

(ii) the alkali is any one of DIPEA, DIEA, TEA and NMM;

(iii) the organic solvent is selected from any one of DMF, DCM, 1,4-dioxane and THF;

(iv) the condensing agent is selected from any one of HATU, HBTU, HCTU, TBTU, TSTU and TNTU;

(v) the reaction temperature of the reaction is 0-50 ℃.

8. The method for preparing pyrimidine tankyrase 2 inhibitor according to claim 3 or 7, wherein in step (4), the acid corresponding to the R group has the structural formula:

wherein R is₁Is selected from

Any one of them.

9. The method for preparing pyrimidine tankyrase 2 inhibitor according to claim 8, wherein R is₁Is selected from

10. Use of the pyrimidine tankyrase 2 inhibitor of claim 1 or 2 in the preparation of a medicament for inhibiting tankyrase 2.