CN113248474A

CN113248474A - Five-membered azole heterocyclic derivative and preparation method and application thereof

Info

Publication number: CN113248474A
Application number: CN202110563258.2A
Authority: CN
Inventors: 王能能; 李妍
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-05-24
Filing date: 2021-05-24
Publication date: 2021-08-13

Abstract

The invention discloses a five-membered azole heterocyclic derivative, and discloses a preparation method and application thereof. The invention belongs to the technical field of medicines, and the penta-azole heterocyclic derivative provided by the invention reduces the cytotoxicity of the compound, and improves the cell activity, oral absorption rate and bioavailability of the medicine; the compound of the invention has the advantages of low cytotoxicity, good cell activity, high oral absorbability and good bioavailability. In addition, it is a further object of the present invention to provide such compounds and pharmaceutically acceptable salts and compositions thereof which are useful for treating a variety of diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport. The compounds provided by the present invention are also useful for the study of nuclear transport modulation in biological and pathological phenomena, such as the study of kinase-mediated intracellular signal transduction pathways and comparative evaluation of novel nuclear transport modulators.

Description

Five-membered azole heterocyclic derivative and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a five-membered azole heterocyclic derivative, and a preparation method and application thereof.

Background

The nucleus of eukaryotic cell has double-layer membrane structure, small molecular protein, nucleic acid and salt in cytoplasm diffuse through nuclear pore, and nuclear mass transfer receptor mediates macromolecular substance to carry out trans-nuclear membrane transfer. Specific proteins and RNAs are transported into and out of the cell nucleus by specific transport molecules, wherein proteins that transport molecules out of the cell nucleus are classified as nuclear export proteins. Nuclear export protein 1(exportin 1, XPO1), also known as chromosomal region stabilizing protein 1(CRM1), is a component of the Golgi apparatus, is one of the most important nuclear export receptors of the karyopherin-beta family, and is the major nuclear receptor for export proteins. Nuclear export protein (XPO-1) is a key nuclear cytoplasmic transporter in cells responsible for the transport of proteins (including tumor suppressor proteins) out of the nucleus, and nuclear export of tumor suppressor proteins is an important mechanism for tumor cells to escape apoptosis. In tumor cells, due to overexpression of XPO-1, which makes this tightly controlled protein transport deregulated, part of the cargo proteins of XPO-1, such as tumor suppressors, are abnormally localized in the cytoplasm, leading to the development and progression of tumors. High expression in several kinds of solid tumor cells and blood tumor cells (such as human ovarian cancer, cervical cancer, pancreatic cancer, hepatocellular carcinoma, leukemia and myeloma) is one of the important targets for developing antitumor drugs.

Nuclear export protein 1(exportin 1, XPO1), responsible for the nuclear transport of more than 240 proteins, including various Tumor Suppressor Proteins (TSPs), such as P53, P73, FOXO1, etc.; growth Regulatory Proteins (GRPs), such as IkB, Rb, P21, P27, BRCA1, APC, etc.; and anti-apoptotic proteins such as NPM, survivin and AP-1, and the like. In the nucleus, XPO1 and RanGTP are combined with cargo protein containing NES (ideal-rich nuclear export signal) to form a stable nuclear transport complex, and the stable nuclear transport complex passes through the central channel of the nuclear pore complex through the interaction of XPO1 and nuclear pore protein; after entering cytoplasm, RanGTP is hydrolyzed into RanGDP under the action of RanGAP, the nuclear transport complex is depolymerized, and the cargo protein is released. The first generation XPO1 inhibitor selinexor showed high blood brain barrier penetration and toxicity problems in preclinical and clinical studies. A common side effect is myelosuppression, with studies reporting patients experiencing emesis, nausea, fatigue, diarrhea, constipation, upper respiratory infections, and low blood sodium levels (hyponatremia). In response to these demanding clinical needs, the development of a new generation of XPO1 inhibitors provides patients with more innovative drugs to achieve better clinical benefits.

Disclosure of Invention

Aiming at the situation, in order to make up for the existing defects, the invention provides a five-membered azole heterocyclic derivative, a preparation method and application thereof.

The invention provides the following technical scheme: the five-membered azole heterocyclic derivative is a compound shown in a formula I, or an optical isomer, an enantiomer, a diastereomer, a racemate or a racemic mixture thereof, or a solvate, a prodrug thereof, or a pharmaceutically acceptable salt thereof;

further, R1 is independently optionally selected from substituted C5-C15 heteroaryl;

r1 is an optionally substituted 5-6 membered heteroaryl having 1,2, or 3 heteroatoms independently selected from the group consisting of: nitrogen, oxygen and sulfur.

Further, R1 is independently optionally selected from substituted pyrrolyl, furanyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, or oxadiazolyl.

Further, R1 is independently optionally selected from unsubstituted or substituted pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl or triazinyl.

Further, R1 is an optionally substituted 8-15 membered fused heteroaryl having 1,2, 3, or 4 heteroatoms independently selected from the group consisting of: nitrogen, oxygen and sulfur.

Further, R1 is independently selected from unsubstituted or substituted indole, isoindole, benzopyrazole, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, isoquinoline, quinoline, chromone, coumarin, and flavone.

Further, R2, R3, R4 are independently selected from hydrogen, unsubstituted or substituted alkyl, alkoxy, hydroxy, amino, halogen, cyano, unsubstituted or substituted ester, unsubstituted or substituted carbonyl compound.

Use of penta-azole heterocyclic derivatives for the treatment of a variety of diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.

The invention with the structure has the following beneficial effects: compared with the prior art, the penta-azole heterocyclic derivative provided by the invention improves the cell activity, oral absorption rate and bioavailability of the medicament; more specifically, the compound of the present invention has the advantages of good cell activity, high oral absorbability, good bioavailability, etc., so the compound of the present invention has better drug-forming properties.

In addition, it is a further object of the present invention to provide such compounds and pharmaceutically acceptable salts and compositions thereof which are useful for treating a variety of diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport. The compounds provided by the present invention are also useful for the study of nuclear transport modulation in biological and pathological phenomena, such as the study of kinase-mediated intracellular signal transduction pathways and comparative evaluation of novel nuclear transport modulators.

Detailed Description

The technical solution of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments; all other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1:

step 1.1: preparation of Compound 1

3-bromo-1, 2, 4-triazole (4.44g, 30.0mmol) was dissolved in 30ml of DMF and triethylenediamine (6.73g, 60.0mmol) was added, followed by stirring at room temperature for 30 minutes. Then (2Z) -isopropyl-3-iodoprop-2-enoate (7.92g, 33.0mmol) was added and stirred at room temperature for 1 h. The mixture was poured into 100ml of ice water and the aqueous phase was extracted with ethyl acetate (3 × 50 ml). The combined organic phases were washed twice with 50ml brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. Column chromatography gave the title product (6.25g, 24.0mmol, 80% yield) as an off-white solid.

t_R＝1.222min,MS(ESI)m/z＝259.8,261.7[M+H]⁺.

Step 1.2: preparation of Compound 2

Isopropyl (Z) -3- (3-bromo-1H-1, 2, 4-triazol-1-yl) acrylate (3.6g, 13.8mmol) was dissolved in dichloromethane (50ml), and bromine (4.41g, 27.6mmol) was slowly added dropwise at 0 ℃. The mixture was stirred at room temperature overnight. After the reaction was complete, the mixture was poured into ice water (50mL) and extracted with dichloromethane (2X 50 mL). The organic phase was washed with saturated brine (3 × 50mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude product as a colorless oil (5.12g, 12.2mmol, 88% yield). It was used directly in the next step without any purification.

t_R＝1.334min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝419.4,421.4[M+H]⁺.

Step 1.3: preparation of Compound 3

Dissolve isopropyl (Z) -2-bromo-3- (3-bromo-1H-1, 2, 4-triazol-1-yl) acrylate (5.12g, 12.2mmol) in tetrahydrofuran (50mL) and add triethylamine (2.47g, 24.4mmol) at 0 deg.C. The mixture was stirred at room temperature for 15 h. The mixture was diluted with 50ml water, the aqueous layer was extracted with 3 × 50ml of ethyl acetate, then the organic phase was washed twice with 50ml of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product as a pale yellow solid (3.4g, 10.0mmol, 82% yield). The crude product was used in the next reaction without further purification.

t_R＝1.280,1.313min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝339.5[M+H]⁺.

Step 1.4: preparation of Compound 4

(Z) -2-bromo-3- (3-bromo-1H-1, 2, 4-triazol-1-yl) acrylic acid isopropyl ester (1017mg, 3mmol) was dissolved in THF/H₂O (8ml/8 ml). Water and lithium hydroxide (251.76mg, 6mmol) were added and cooled to 0 deg.C, the reaction mixture was stirred at 0 deg.C for 1h, the reaction was concentrated in vacuo and the residue was diluted with 10ml of water. Then it was adjusted to pH 3 with concentrated hydrochloric acid, precipitation appeared, and the precipitate was collected by filtration and washed with water (5 mL). The solid was the title product (280mg, 0.95mmol, 31.5% yield) as an off-white solid.

t_R＝0.970min,MS(ESI)m/z＝297.6[M+H]⁺.

¹H NMR(400MHz,DMSO-d₆)δ14.01(brs,1H),9.22(s,1H),8.78(s,1H).

Step 1.5: preparation of Compound 5

(2Z) -2-bromo-3- (3-bromo-1, 2, 4-triazol-1-yl) prop-2-enoic acid (240mg, 0.8mmol) was dissolved in 5mL THF and isopropyl chloride (196mg, 1.6mmol) and carminorphine (121mg, 1.2mmol) were added at 0 degrees. The reaction mixture was stirred at 0 ℃ for 1 hour, then ammonia/methanol solution (3mL,7M) was added, and stirring was continued for 30 minutes. The resulting mixture was concentrated in vacuo, the residue diluted with 10ml ice water, the aqueous phase extracted with ethyl acetate 3 × 20ml and the organic phase washed with brine, dried over anhydrous sodium sulfate and concentrated in vacuo to afford the title product (200mg, 0.68mmol, 84% yield) as an off-white solid.

t_R＝0.971min,MS(ESI)m/z＝296.6[M+H]⁺.

¹H NMR(400MHz,DMSO-d6)δ9.18(s,1H),8.56(s,1H),7.95～7.93(m,2H).

Step 1.6: preparation of Compound Int-1

(2Z) -2-bromo-3- (3-bromo-1, 2, 4-triazol-1-yl) prop-2-enamide (480mg, 1.62mmol), pyrimidine 5-borate (301mg, 2.43mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (118.54mg, 0.16mmol) and potassium acetate (318g, 3.24mmol) were added to 10/1mL dioxane/H2O solution. Stirring was carried out at 80 ℃ for 2 hours under nitrogen. The reaction solution was concentrated. Then, it was put on a silica gel column and eluted with (EA: MeOH 30:1) to give the objective product (240mg, 0.81mmol, 50% yield) as a red solid.

t_R＝0.835min,MS(ESI)m/z＝294.7,296.7[M+H]⁺.

¹H NMR(400MHz,DMSO-d6)δ9.17(s,1H),8.91(s,1H),8.64(s,2H),8.25(s,1H).

Step 1.7: preparation of Compound P-A1

(E) -3- (3-bromo-1H-1, 2, 4-triazol-1-yl) -2- (pyrimidin-5-yl) acrylamide (Int-1) (80mg, 0.27mmol), [ 3-chloro-5- (trifluoromethyl) phenyl ] boranediol (SM3) (73mg, 0.33mmol) and potassium carbonate (74.94mg,0.54mmol) were dissolved in dioxane/water (11mL, v/v ═ 10:1), and tetrakis (triphenylphosphine) palladium (31.33mg,0.027mmol) was added under nitrogen. The reaction mixture was stirred at 80 ℃ for 10h, then diluted with water (10mL) and extracted with ethyl acetate (3X 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. This was purified by prep-TLC (EA as eluent) and prep-HPLC to give (E) -3- (3-chloro-5- (trifluoromethyl) phenyl) -1H-1,2, 4-triazol-1-yl) -2- (pyrimidin-5-yl) acrylamide (P-A1) (21.8mg, 20.18% yield) as a white solid.

t_R＝1.184min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝394.7[M+H]⁺.

HPLC:t_R＝5.989min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).¹H NMR(300MHz,DMSO-d6)δ9.19(s,1H),9.08(s,1H),8.69(s,2H),8.38(s,1H),7.89(d,J＝40.2Hz,2H),7.65(d,J＝16.7Hz,2H),7.37(s,1H).

The example compounds of the following table 1 were prepared according to the same method as the above examples, using a commercially available compound or a preparation method referring to intermediate compounds shown.

[ TABLE 1 ]

Example 2: preparation of Compound P-B1

Preparation of intermediate Int-2

Step 1.1: preparation of Compound 1

t_R＝1.222min,MS(ESI)m/z＝259.8,261.7[M+H]⁺.

Step 1.2: preparation of Compound 2

Step 1.3: preparation of Compound 3

Step 1.4: preparation of Compound 4

t_R＝0.970min,MS(ESI)m/z＝297.6[M+H]⁺.

¹H NMR(400MHz,DMSO-d₆)δ14.01(brs,1H),9.22(s,1H),8.78(s,1H).

Step 1.5: preparation of Compound 5

t_R＝0.971min,MS(ESI)m/z＝296.6[M+H]⁺.

¹H NMR(400MHz,DMSO-d6)δ9.18(s,1H),8.56(s,1H),7.95～7.93(m,2H).

Step 1.6: preparation of Compound Int-2

(2Z) -2-bromo-3- (3-bromo-1, 2, 4-triazol-1-yl) prop-2-enamide (480mg, 1.62mmol), quinoline-3-boronic acid (420mg, 2.43mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (118.54mg, 0.16mmol) and potassium acetate (318g, 3.24mmol) were added to 10/1mL dioxane/H2O solution. Stirring was carried out at 80 ℃ for 2 hours under nitrogen. The reaction solution was concentrated. The column was then loaded with silica gel to give the desired product (300mg, 53% yield).

t_R＝1.105min,MS(ESI)m/z＝344.1,346.2[M+H]⁺.

Step 1.7: preparation of Compound P-B1

Compound Int-2(93mg, 0.27mmol), [ 3-chloro-5- (trifluoromethyl) phenyl ] boranediol (SM3) (73mg, 0.33mmol) and potassium carbonate (74.94mg,0.54mmol) were dissolved in dioxane/water (11mL, v/v ═ 10:1), and tetrakis (triphenylphosphine) palladium (31.33mg,0.027mmol) was added under nitrogen. The reaction mixture was stirred at 80 ℃ for 10h, then diluted with water (10mL) and extracted with ethyl acetate (3X 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. It was purified by prep-TLC (EA as eluent) and prep-HPLC to give the desired product (P-B1) (23.8mg, yield 20.18%) as a white solid.

t_R＝1.284min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝444.2[M+H]⁺.

HPLC:t_R＝6.989min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1 x50 mm,3.5um) the example compounds of table 2 below were prepared according to the same method as the above examples, using commercially available compounds or referring to the preparation method of the intermediate compounds shown.

[ TABLE 2 ]

Example 3: cell antiproliferative activity (IC)₅₀) Testing

Cells were prepared, 100uL of cells were added to a 96-well plate, and incubated overnight at 37 ℃. The next day the cells were diluted in DMSO to make up the desired final concentration. The samples were diluted 3-fold to the desired concentration and 50uL of the sample was added to the cell culture medium at 37 ℃ with 5% CO₂And (5) incubating for 72 h. After equilibration to room temperature, 40uL of the suspension was removed

Reagent is added into the pore plate, mixed evenly, incubated for 60 minutes at room temperature and then detected.

[ TABLE 3 ] measurement results of the Compounds

In table 3: a is less than or equal to 30nM, B is less than or equal to 30nM and less than or equal to 100nM, C is less than or equal to 1 muM and 100nM, D is more than 1 muM, and NT is not tested

The data in the table show that the compound of the invention has stronger anti-proliferation activity of tumor cells (human multiple myeloma cells (MM.1S)), for example, the compounds P-A1 and P-B1 have higher anti-tumor activity of human multiple myeloma cells MM.1S than positive drug KPT-8602, and meanwhile, the inhibition concentration of P-A1 and P-B1 on mouse embryo fibroblasts (3T3) is higher, and the compound has higher safety than the positive drug KPT-8602.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is represented by any one of the following structural formulas:

2. the compound of claim 1, wherein the pharmaceutically acceptable salt comprises: a salt of a compound represented by the general formula (1) with an acid; wherein the acid comprises inorganic acid, organic acid and acidic amino acid; the inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid; the organic acid is selected from formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, p-toluenesulfonic acid, ethanesulfonic acid or benzenesulfonic acid; the acidic amino acid is selected from aspartic acid or glutamic acid.

3. A pharmaceutical composition for the treatment, modulation and/or prevention of diseases associated with XPO 1-related physiological conditions, characterized in that it contains a pharmaceutically acceptable excipient or carrier and, as active ingredient, a compound according to any one of claims 1-2, or each optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof.

4. The pharmaceutical composition of claim 3, wherein said composition is in an oral dosage form.

5. Use of a compound according to any one of claims 1-2, or each optical isomer, each crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof, for the preparation of a pharmaceutical composition for the treatment, modulation and/or prevention of a disease associated with a physiological condition associated with XPO 1.

6. Use of a compound according to any one of claims 1-2, or each optical isomer, each crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof, for the manufacture of a medicament for the treatment, modulation and/or prevention of a disease associated with a physiological condition associated with XPO 1.