WO2024087977A1 - Phenylurea compound, and preparation method therefor, use thereof and pharmaceutical composition thereof - Google Patents

Abstract

The present invention relates to a phenylurea compound capable of targeting SHP2, and a preparation method therefor, the use thereof and a pharmaceutical composition thereof. The compound is as represented by formula (I). The present invention further relates to a series of phenylurea SHP2 allosteric inhibitors having high SHP2 selective activity, which have a novel structure and are expected to solve the current situation of the lack of marketed inhibitor drugs for SHP2 at present.

Description

Phenylurea compounds and preparation methods, uses and pharmaceutical compositions thereof Technical Field

The present invention relates to a compound and a preparation method, use and pharmaceutical composition thereof, in particular to a phenylurea compound and a preparation method, use and pharmaceutical composition thereof.

Background technique

Protein tyrosine phosphatases (PTPs) can participate in a variety of cellular activities, including proliferation, differentiation, metabolism, and immune-related responses, by catalyzing the dephosphorylation of substrates. Abnormal tyrosine phosphorylation activity of body proteins has been shown to be associated with the occurrence and development of a variety of human diseases, including cancer, autoimmune dysfunction, etc. Currently, increased tyrosine phosphorylation activity has become a hallmark of many cancers.

SHP2 is a non-receptor protein tyrosine phosphatase containing the Src homology 2 domain encoded by PTPN11 and consists of 593 amino acid residues. SHP2 is located in the cytoplasm and is induced by various tyrosine kinases to participate in a variety of intracellular carcinogenic signaling pathways, such as PI3K/AKT, RAS/Raf/MARK and PD-1/PD-L1. SHP2 is one of the important targets for cancer treatment. The gain-of-function mutation encoded by PTPN11 of SHP2 is found in a variety of sporadic solid tumors. In addition, the overactivation of SHP2 also plays an important role in the development of cancer. Almost all RTK inhibitors activate the RAS pathway through SHP2. By blocking or inhibiting the activation of the SHP2 pathway, the effect of tumor treatment can be significantly improved. Therefore, SHP2 inhibitors have the potential to become broad-spectrum anti-tumor drugs.

At present, the development of SHP2 inhibitors is mainly divided into two categories. One is type I SHP2 inhibitors based on the catalytic site of the PTP domain. This type of inhibitor has a highly conservative sequence of the catalytic site and general selectivity. Because this type of inhibitor has a strong electron-withdrawing group, it has poor membrane permeability and low oral bioavailability. The second type is SHP2 allosteric inhibitors, which were first reported by Novartis in 2016 (Nature, 2016, 535, 148-152.). This type of allosteric inhibitor binds to the cavity pocket formed by the C-SH2, N-SH2 and PTP domains of the SHP2 protein, hindering the tyrosine phosphorylation substrate from entering the catalytic site and inhibiting the activity of the SHP2 protein.

Although important breakthroughs have been made in the research of SHP2 small molecule inhibitors, the development of related SHP2 inhibitors is still in the early clinical research stage, and no inhibitor targeting SHP2 has been successfully marketed. In view of the defects of low inhibitor selectivity and poor bioavailability caused by the high homology of the catalytic domain of the PTPs family, it is now urgent to further develop new SHP2 allosteric inhibitors with high SHP2 selectivity and drugability.

Summary of the invention

Purpose of the invention: The purpose of the present invention is to provide a series of phenylurea compounds with high SHP2 selective activity; another purpose of the present invention is to provide a method for preparing the above phenylurea compounds ... One object is to provide the application of the above-mentioned phenylurea compounds; another object of the present invention is to provide a pharmaceutical composition containing heterocyclic ether compounds.

Technical solution: The phenylurea compound of the present invention has the general formula I as shown:

in:

R1 and R2 are selected from hydrogen, deuterium, substituted or unsubstituted C _1-10 alkyl, hydroxyl; the substituent of the substituted C _1-10 alkyl is selected from one or more of the following groups: amino, hydroxyl, carboxyl, amide, cyano, olefin, alkyne, C _1-6 alkoxy;

R3, R4, R5 and R6 are each the same or different and are independently selected from hydrogen, deuterium, halogen, substituted or unsubstituted amino, hydroxyl, substituted C _1-6 alkoxy, substituted or unsubstituted C _1-10 alkyl; the substituents of the substituted amino, substituted C _1-10 alkyl, and substituted C _1-6 alkoxy are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C _1-10 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl;

p, q, s and t are independently selected from 0, 1, 2, 3;

W is selected from an oxygen atom or a sulfur atom;

X ₁ , X ₂ and X ₃ are independently selected from a carbon atom or a nitrogen atom;

Y ₁ , Y ₂ , Z ₁ and Z ₂ are independently selected from a carbon atom, an oxygen atom or a nitrogen atom;

_Z3 and _Z4 are independently selected from a carbon atom, an oxygen atom, a nitrogen atom or a hydrogen atom;

Ring A is selected from a benzene ring, a 3-6 membered heterocycle, a benzene ring and a 3-6 membered heterocycle, a 6 membered heterocycle and a 3-6 membered heterocycle;

Ring B is selected from hydrogen, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl; the C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl may be substituted by one or more of the following groups: halogen, hydroxyl, carboxyl, amide, cyano, nitro, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _1-6 alkoxy, substituted or unsubstituted amino; the substituted C _1-10 alkyl, substituted C _1-6 alkoxy, substituted amino are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C _1-6 alkoxy, C _1-10 alkyl;

Ring C is hydrogen or C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl including Z3 and Z4; the C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, The 3-10 membered heteroaryl and C _6-10 aryl may be substituted by one or more of the following groups: halogen, hydroxyl, carboxyl, amide, cyano, nitro, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _1-6 alkoxy, substituted or unsubstituted amino; the substituted C _1-10 alkyl, substituted C _1-6 alkoxy, substituted amino are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, olefin, alkyne, C _1-6 alkoxy, C _1-10 alkyl;

Preferably, in the phenylurea compound, R1 and R2 are independently selected from H, methyl or ethyl;

The R4 is independently selected from

The R5 is independently selected from Methyl or

The R6 is independently selected from a chlorine atom, a fluorine atom, and a methyl group;

X1 is N, X2 is N, and X3 is C;

Y1, Y2 is C;

Z1, Z2 is C;

Z3, Z4 are C or hydrogen;

n, m are independently selected from 0 or 1;

p is 1, q is 0, s is 0 or 2, t is 1 or 2;

Ring A is selected from:

Ring B is selected from: a 6-membered aromatic heterocycle, a benzene ring or a C _3-8 cycloalkyl group; the 6-membered aromatic heterocycle, the benzene ring or the C _3-8 cycloalkyl group may be substituted by one or more of the following groups: chlorine, fluorine, bromine, methoxy, carboxyl, nitro, cyano or amino.

Preferably, the phenylurea compound includes a pharmaceutically acceptable salt, racemate, optical isomer or solvate thereof.

Preferably, the phenylurea compound comprises any one of the following structures:

On the other hand, the present invention provides a method for preparing the above-mentioned phenylurea compound, when R1 and R2 are independently selected from H, methyl or ethyl;

The R4 is independently selected from

The R5 is independently selected from Methyl or

The R6 is independently selected from a chlorine atom, a fluorine atom, and a methyl group;

X1 is N, X2 is N, and X3 is C;

Y1, Y2 is C;

Z1, Z2 is C;

Z3, Z4 are C or hydrogen;

p is 1, q is 0, s is 0 or 2, t is 1 or 2;

Ring A is selected from:

Ring B is selected from: a 6-membered aromatic heterocyclic ring, a benzene ring or a C _3-8 cycloalkyl group;

The synthetic route of the phenylurea compound is shown in the following formula:

The specific synthesis steps are as follows:

1) Compound H2 reacts with carbon disulfide or triphosgene to obtain compound H3;

2) H1 and H3 are condensed to give compound H4.

Wherein, the solvent used in step 1) includes but is not limited to: chloroform, tetrahydrofuran, dichloromethane, acetone, acetonitrile, N,N-dimethylformamide or a mixed solvent optionally composed of these solvents;

The solvent used in step 2) includes, but is not limited to: chloroform, tetrahydrofuran, dichloromethane, acetone, acetonitrile, N,N-dimethylformamide or a mixed solvent optionally composed of these solvents;

On the other hand, the present invention provides an application of the above-mentioned phenylurea compounds in the preparation of inhibitors having SHP2 inhibitory activity, wherein the phenylurea compounds include substituted phenylurea compounds and pharmaceutically acceptable salts, racemates, optical isomers or solvent compounds thereof.

On the other hand, the present invention provides an application of the above-mentioned phenylurea compound in the preparation of an anti-tumor drug, wherein the phenylurea compound includes a pharmaceutically acceptable salt, racemate, optical isomer or solvent compound thereof.

On the other hand, the present invention provides a pharmaceutical composition comprising the above-mentioned phenylurea compound, wherein the composition is composed of the phenylurea compound or its pharmaceutically acceptable salt, racemate, optical isomer or solvent compound as an active ingredient and a pharmaceutically acceptable carrier.

The pharmaceutical composition containing the phenylurea compound is a capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhalant, ointment, suppository or patch.

Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The present invention provides a series of phenylurea-type SHP2 allosteric inhibitors with high SHP2 selective activity, which have novel structures and are expected to solve the current situation that there are no SHP2 inhibitor drugs on the market.

Detailed ways

The present invention will be further described below with reference to the embodiments.

Intermediate 1

The synthesis of intermediate 1 was prepared according to J.Med.Chem.2020,63,13578-13594.

Intermediate 2

Intermediate 2 was prepared by referring to the synthesis method of J.Med.Chem.2020,63,13578-13594.

Intermediate 3

The synthesis of intermediate 3 was prepared by referring to the synthesis method of patent CN112839935.

Example 1

synthetic route:

Synthesis of Compound 1-B

500 mg of compound 1-A, 1 g of tert-butyl mercaptan and 2.8 g of cesium carbonate were dissolved in 15 mL of DMF, protected by nitrogen, and stirred at 120°C for one day. 50 mL of water was added, extracted twice with 100 mL of ethyl acetate, washed once with saturated brine, and the organic phase was collected and concentrated to obtain 550 mg of compound 1-B.

Synthesis of Compound 1-C

550 mg of compound 1-C was dissolved in 10 mL of concentrated hydrochloric acid and stirred at 80° C. After 4 hours, TLC monitored that the reaction of the raw material was complete, and the solvent was dried with an oil pump to obtain compound 1-C.

Synthesis of Compound 1-D

Dissolve 200 mg of compound 1-C, 530 mg of potassium phosphate and 200 mg of 2-amino-3-bromo-6-chloropyrazine in 1,4-dioxane and stir at room temperature for 15 minutes. Add 38 mg of cuprous iodide and 72 mg of 1,10-phenanthroline, protect with nitrogen, stir and react overnight at 90°C. Monitor the reaction by TLC, spin dry and column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 1-D.

Synthesis of Compound 1-E

100 mg of compound 1-D and 150 mg of tert-butyl (4-methylpiperidin-4-yl)carbamate were dissolved in DMSO, 174 μL of DIPEA was added, and the mixture was stirred at 100°C for reaction. After five hours, the reaction was completed by TLC monitoring, 50 mL of water was added, and the mixture was extracted twice with 100 mL of ethyl acetate, washed once with saturated brine, and the organic phase was collected and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 1-E.

Synthesis of Compound 1-F

80 mg of compound 1-E and 35 mg of phenyl isocyanate were dissolved in THF and stirred at 60° C. After four hours, the reaction was completed by TLC monitoring, and the mixture was concentrated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain 90 mg of compound 1-F.

Synthesis of compound 1

90 mg of compound 1-F was dissolved in 1.5 mL of dichloromethane and 0.5 mL of trifluoroacetic acid, and the mixture was stirred at room temperature for half an hour. The reaction was completed after monitoring by TLC. The solvent was dried and purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 28 mg of white solid compound 1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.76 (s, 1H), 8.50 (s, 1H), 7.89 (d, J=8.3 Hz, 1H), 7.75 (s, 1H), 7.33 (t, J=8.0 Hz, 2H), 7.29-7.22 (m, 2H), 7.15 (t, J=8.1 Hz, 1H), 7.05 (dd, J=7.8, 1.9 Hz, 1H), 6.31 (dd, J=8.0, 1.4 Hz, 1H), 6.18 (s, 2H), 3.42-3.34 (m, 4H), 1.69 (t, J=5.7 Hz, 4H), 1.34 (s, 3H).

Intermediate 4

Synthesis of Intermediate 4 The intermediate 4 can be prepared by referring to the synthesis method of 1-E in Example 1.

Using the above method, the following examples can be synthesized with the corresponding starting materials:

Embodiment 18

synthetic route:

Synthesis of compound 2-B

Compound 2-B can be prepared by the synthesis method of reference J. Med. Chem. 2011, 54, 7066-7083.

Synthesis of compound 18

Referring to the synthesis method of compound 1, compound 18 can be prepared. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.65 (s, 1H), 8.47 (s, 1H), 8.27 (s, 1H), 7.15 (t, J = 8.1 Hz, 1H), 7.01 (dd, J = 7.4, 2.0 Hz, 1H), 6.53 (dd, J = 7.8, 1.5 Hz, 1H), 6.22 (s, 1H), 4.05-3.96 (m, 4H), 1.86 (t, J = 5.5 Hz, 4H), 1.34 (s, 3H).

Using the above two synthesis methods, the following examples can be synthesized with the corresponding starting materials:

Embodiment 40

tablet

Compound 1 (50 g) prepared in Example 11, hydroxypropylmethylcellulose E (150 g), starch (200 g), appropriate amount of povidone K30 and magnesium stearate (1 g) were mixed, granulated and tableted.

In addition, according to the conventional preparation method of the Pharmacopoeia 2015 edition, the compounds prepared in Examples 1-66 can be added with different pharmaceutical excipients to prepare capsules, powders, granules, pills, injections, syrups, oral solutions, inhalants, ointments, suppositories or patches, etc.

Test Example 1

Pharmacological experiments have shown that the SHP2 inhibitory activity of the present invention can be used to prepare anti-tumor drugs. The following are the pharmacological experimental results of some compounds of the present invention:

Determination of the inhibitory effect of pharmacological experimental compounds on PD-1/PD-L1 interaction:

(I) Reagents and consumables

(II) Instruments

Centrifuge (manufacturer: Eppendorf, model: 5430)

Microplate reader (Manufacturer: Perkin Elmer, Model: EnSight)

Echo 550 (Manufacturer: Labcyte, Model: Echo 550)

(III) Experimental methods

(1) Prepare 1×Reaction buffer.

(2) Preparation of compound concentration gradient: The test compound concentration is 10000nM, 3-fold dilution, 10 concentration points, single-well detection. Dilute to 100-fold final concentration solution in a 384-well plate, and then transfer 200nL to a 384-well reaction plate using Echo550 for later use. Add 200nL of 100% DMSO to the negative control well and the positive control well, respectively.

(3) Use 1× reaction buffer to prepare an activation peptide solution with a 5-fold final concentration, add 5 μL to each reaction plate, and centrifuge at 1000 rpm for 1 min.

(4) Use 1× reaction buffer to prepare an enzyme solution with a final concentration of 2.5 times, add 10 μL to each reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 60 min.

(5) Use 1× reaction buffer to prepare a substrate solution with a final concentration of 2.5 times, add 10 μL to the reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 20 min.

(6) Use EnSight to read the Ex355/Em460 fluorescence values.

(IV) Data Analysis

Calculation formula: %Inhibition = (Mean(PC)-RFU)/(Mean(PC)-Mean(NC))*100

Where: RFU: fluorescence value of the sample; Mean(NC): mean fluorescence value of the control well containing 10μM SHP099. Mean(PC): mean fluorescence value of the positive control well.

Fitting dose-effect curve

The log value of the concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism5 was used to fit the dose-effect curve to obtain the IC50 value of each compound on the enzyme activity.

(V) Experimental results

The following table shows the activity range or IC ₅₀ of the compounds against SHP2 inhibition. The ranges are as follows: A = 1 nM-10 nM; B = 10.01 nM-100 nM; C = 100.1-1000 nM.

Claims (9)

Phenylurea compounds having the general formula I
in: R1 and R2 are selected from hydrogen, deuterium, substituted or unsubstituted C _1-10 alkyl, hydroxyl; the substituent of the substituted C _1-10 alkyl is selected from one or more of the following groups: amino, hydroxyl, carboxyl, amide, cyano, olefin, alkyne, C _1-6 alkoxy; R3, R4, R5 and R6 are each the same or different and are independently selected from hydrogen, deuterium, halogen, substituted or unsubstituted amino, hydroxyl, substituted C _1-6 alkoxy, substituted or unsubstituted C _1-10 alkyl; the substituents of the substituted amino, substituted C _1-10 alkyl, and substituted C _1-6 alkoxy are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C _1-10 alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl; p, q, s and t are independently selected from 0, 1, 2, 3; W is selected from an oxygen atom or a sulfur atom; X ₁ , X ₂ and X ₃ are independently selected from a carbon atom or a nitrogen atom; Y ₁ , Y ₂ , Z ₁ and Z ₂ are independently selected from a carbon atom, an oxygen atom or a nitrogen atom; _Z3 and _Z4 are independently selected from a carbon atom, an oxygen atom, a nitrogen atom or a hydrogen atom; Ring A is selected from a benzene ring, a 3-6 membered heterocycle, a benzene ring and a 3-6 membered heterocycle, a 6 membered heterocycle and a 3-6 membered heterocycle; Ring B is selected from hydrogen, C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl; the C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl may be substituted by one or more of the following groups: halogen, hydroxyl, carboxyl, amide, cyano, nitro, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _1-6 alkoxy, substituted or unsubstituted amino; the substituted C _1-10 alkyl, substituted C _1-6 alkoxy, substituted amino are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C _1-6 alkoxy, C _1-10 alkyl; Ring C is hydrogen or C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl including Z3 and Z4; the C _3-8 cycloalkyl, C _3-8 cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heteroaryl, C _6-10 aryl may be substituted by one or more of the following groups: halogen, hydroxyl, carboxyl, amide, cyano, nitro, substituted or unsubstituted C _1-10 alkyl, substituted or unsubstituted C _1-6 alkoxy, substituted or unsubstituted amino; the substituted C _1-10 alkyl, substituted C _1-6 alkoxy, substituted amino are independently selected from one or more of the following groups: halogen, amino, hydroxyl, carboxyl, amide, cyano, alkene, alkyne, C _1-6 alkoxy, C _1-10 alkyl. The phenylurea compound according to claim 1, characterized in that: Said R1, R2 are independently selected from H, methyl or ethyl; The R4 is independently selected from The R5 is independently selected from Methyl or The R6 is independently selected from a chlorine atom, a fluorine atom, and a methyl group; X1 is N, X2 is N, and X3 is C; Y1, Y2 is C; Z1, Z2 is C; Z3, Z4 are C or hydrogen; p is 1, q is 0, s is 0 or 2, t is 1 or 2; Ring A is selected from: Ring B is selected from: a 6-membered aromatic heterocycle, a benzene ring or a C _3-8 cycloalkyl group; the 6-membered aromatic heterocycle, the benzene ring or the C _3-8 cycloalkyl group may be substituted by one or more of the following groups: chlorine, fluorine, bromine, methoxy, carboxyl, nitro, cyano, amino. The phenylurea compound according to claim 1, characterized in that the compound comprises any one of the following structures:

The phenylurea compound according to claim 1, characterized in that the compound includes a pharmaceutically acceptable salt, tautomer, mesomer, racemate, stereoisomer, prodrug or solvent compound thereof; A method for preparing a phenylurea compound according to any one of claims 1 to 3, characterized in that when R1 and R2 are independently selected from H, methyl or ethyl; The R4 is independently selected from The R5 is independently selected from Methyl or The R6 is independently selected from a chlorine atom, a fluorine atom, and a methyl group; X1 is N, X2 is N, and X3 is C; Y1, Y2 is C; Z1, Z2 is C; Z3, Z4 are C or hydrogen; p is 1, q is 0, s is 0 or 2, t is 1 or 2; Ring A is selected from: Ring B is selected from: a 6-membered aromatic heterocyclic ring, a benzene ring or a C _3-8 cycloalkyl group; The synthetic route of the phenylurea compound is shown below:

The specific synthesis steps are as follows: 1) Compound H2 reacts with carbon disulfide or triphosgene to obtain compound H3; 2) H1 and H3 are condensed to give compound H4. Use of the phenylurea compound according to any one of claims 1 to 4 in the preparation of an inhibitor having SHP2 inhibitory activity. A use of the phenylurea compound according to any one of claims 1 to 4 in the preparation of anti-tumor drugs. A pharmaceutical composition containing the phenylurea compound according to any one of claims 1 to 4, characterized in that the pharmaceutical composition is composed of the phenylurea compound or its pharmaceutically acceptable salt, racemate, optical isomer or solvent compound as an active ingredient and a pharmaceutically acceptable carrier. The pharmaceutical composition containing a phenylurea compound according to claim 7, characterized in that the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhalant, ointment, suppository or patch.