CN115894485B - Heteroaryl piperidine derivatives, preparation method thereof and application thereof in medicines - Google Patents

Abstract

The invention relates to heteroaryl piperidine derivatives, a preparation method thereof and application thereof in medicines. Specifically, the invention provides heteroaryl piperidine compounds shown in a formula I, pharmaceutically acceptable salts or stereoisomers thereof and a preparation method thereof. The invention also provides a pharmaceutical composition containing the compound I, and application of the compound I and the pharmaceutical composition thereof in preparing medicines for preventing or treating diseases related to estrogen receptors, in particular to preparing medicines for preventing or treating cancers.

Description

Heteroaryl piperidine derivatives, preparation method thereof and application thereof in medicines

Technical Field

The present invention relates to heteroaryl piperidine derivatives, processes for their preparation and their use as estrogen receptor modulators for the prophylaxis and/or treatment of estrogen receptor mediated or dependent diseases or conditions.

Background

Estrogen receptors include two broad classes: one is the classical nuclear receptor, including erα and erβ, which are located in the nucleus and mediate the genotypic effect of estrogens, i.e., exert a "genotypic" regulatory effect by modulating transcription of specific target genes; second, membrane receptors, including the membrane components of classical nuclear receptors, are GPER1 (GPR 30), gaq-ER and ER-X, which belong to the family of G protein coupled receptors. Estrogen long-term overstimulation, increased levels of erα expression, or enhanced transcriptional activation activity are an important factor in the development of breast cancer.

About 70% of breast cancers express erα, which makes it a major target for the treatment of breast cancers. The current standard of care treatment for ER positive breast cancer patients includes three major classes of drugs: (1) Drugs directed against erα, such as selective estrogen receptor down-regulators (SERDs), selective Estrogen Receptor Modulators (SERMs), and Selective Estrogen Receptor Covalent Antagonists (SERCAs); (2) Aromatase Inhibitors (AIs) that reduce estrogen levels, (3) CDK4/6 kinase inhibitors. Wherein the bonding of SERDs to estrogen receptors not only antagonizes the activity of estrogen receptors, but also promotes the degradation of the receptors.

Chinese patent application No. CN201580069484.8 discloses a class of estrogen receptor modulators, comprising the following compound 340 and compound 277, wherein compound 340 is GDC-9545.GDC-9545 is a potent estrogen receptor degrading agent and is currently in clinical trials.

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Compound 340 (GDC-9545) Compound 277

Because of the low number of estrogen receptor modulators currently on the market, there is a need to develop more estrogen receptor degrading drugs that can be used to treat breast cancer or other estrogen receptor related diseases.

Disclosure of Invention

The invention provides a brand new estrogen receptor modulator, a preparation method thereof and application thereof in preparing medicines for preventing or treating diseases related to estrogen receptors.

The technical scheme of the invention is as follows:

the invention provides a compound shown in a formula I, pharmaceutically acceptable salt or stereoisomer thereof,

I。

in some embodiments, the stereoisomers have the structure shown in formula I-1,

I -1

the invention also provides a pharmaceutical composition comprising compound I, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. Such carriers include adjuvant ingredients conventional in the art, such as, for example, fillers, binders, diluents, disintegrants, lubricants, colorants, flavoring agents, antioxidants, wetting agents, and the like.

The pharmaceutical composition can be prepared into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral liquid, suspension, granules, powder, particles, pills, miniature tablets, instant films, nasal sprays, transdermal patches, injections or various sustained and controlled release preparations and the like. The pharmaceutical composition may be administered orally, transmucosally, rectally or parenterally (including intravascular, intravenous, intraperitoneal, subcutaneous, intramuscular and intrasternal), preferably orally. The dosage to be administered may be appropriately adjusted according to the age, sex and type of disease of the patient.

For oral administration, the pharmaceutical composition may be in the form of, for example, a tablet, capsule, liquid capsule, suspension or solution, or the like. The pharmaceutical compositions are preferably prepared in dosage unit form containing a specific amount of the active ingredient. For example, the pharmaceutical composition may be provided as a tablet or capsule containing about 0.1 to 1000mg, preferably about 0.25 to 250mg, and more preferably about 0.5 to 100mg of the active ingredient. Suitable daily dosages for humans or other mammals can vary widely depending on the condition of the patient and other factors, but can be determined using conventional methods.

The invention also provides a preparation method of the compound I-1 or pharmaceutically acceptable salt thereof, which comprises the following steps:

the invention also provides application of the compound I or I-1, pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition thereof in preparing medicines for preventing or treating diseases related to estrogen receptors. Such estrogen receptor-related diseases include ER- α dysfunction associated with cancer (bone cancer, breast cancer, lung cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, and uterine cancer), central nervous system deficiency (e.g., alcoholism, migraine, etc.), cardiovascular system deficiency (e.g., aortic aneurysm, susceptibility to myocardial infarction, aortic valve sclerosis, cardiovascular disease, coronary artery disease, hypertension, etc.), blood system deficiency, immune and inflammatory diseases, metabolic deficiency (e.g., cholestasis, hypourethral cleavage, obesity, osteoarthritis, osteopenia, osteoporosis, etc.), neurological deficiency (e.g., alzheimer's disease, parkinson's disease, migraine, vertigo, etc.), mental deficiency (e.g., anorexia nervosa, attention deficit hyperactivity disorder, dementia, major depressive disorder, psychosis, etc.), or reproductive deficiency (e.g., menstrual beginner age, endometriosis, infertility), etc.

The invention also provides application of the compound I or I-1, pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition thereof in preparing a medicament for treating cancer. The cancer is selected from breast cancer, lung cancer, ovarian cancer, endometrial cancer, prostate cancer or uterine cancer; breast cancer is preferred. The breast cancer is preferably estrogen receptor positive breast cancer.

The compounds provided by the invention can be used for treating breast cancer alone or in combination with other medicaments for treating breast cancer. Such other drugs include, but are not limited to, aromatase inhibitors, anthracyclines, platinum, nitrogen erucinates, taxanes. Exemplary drugs for treating breast cancer include, but are not limited to, tasilib (GDC-0032), paclitaxel, anastrozole, exemestane, cyclophosphamide, epirubicin, fulvestrant, letrozole, gemcitabine, trastuzumab, polyethylene glycol fegrid, tamoxifen, docetaxel, toremifene, vinorelbine, capecitabine, or ixabepilone.

The compound I or I-1, pharmaceutically acceptable salt or stereoisomer thereof or the pharmaceutical composition thereof provided by the invention has better degradation activity on ER alpha as a selective estrogen receptor degradation agent, and can well inhibit proliferation of breast cancer cells.

Detailed Description

The invention is further illustrated below in conjunction with specific examples and test examples, but is not intended to limit the scope of the invention in any way.

Definition and description

The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless otherwise specifically defined, should not be construed as being ambiguous or otherwise clear, but rather should be construed in a generic sense. When trade names are presented herein, it is intended to refer to their corresponding commercial products or active ingredients thereof.

The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a base in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, organic acid salts, salts of amino acids (e.g., arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the invention contain basic and acidic functionalities that can be converted to either base or acid addition salts.

Pharmaceutically acceptable salts of the invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, the salt is prepared by the following steps: prepared via reaction of these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.

Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are all included within the scope of the present invention.

The compounds of the invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -pairs of enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of the present invention.

Optically active (R) -and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or derivatization with chiral auxiliary wherein the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomeric salt is formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional means well known in the art, and recovery of the pure enantiomer. Furthermore, separation of enantiomers and diastereomers is typically accomplished using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amine).

The present invention is intended to include all isotopes of atoms present in the compounds of the invention. Isotopes include atoms having the same atomic number but different mass numbers. Isotopes of hydrogen include deuterium and tritium as general examples and without limitation. Isotopes of carbon include ¹³ C and C ¹⁴ C. Isotopically-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein using an appropriate isotopically-labeled reagent in place of an otherwise-used non-labeled reagent.

The compounds were named by manual or ChemDraw software and the commercial compounds used the supplier catalog name.

Example 1: synthesis of Compound I-1

The mixture was stirred at 100℃for 16H in a toluene (100 mL) mixture of (R) -1- (1H-indol-3-yl) -N- (2, 2-trifluoroethyl) propan-2-amine (1.5 g,5.9 mmol,1 eq), 2, 6-difluoro-4-hydroxybenzaldehyde (1.1 g,6.4 mmol,1.1 eq) and acetic acid (10 mL) at room temperature. After cooling to room temperature, the mixture was concentrated, the residue was washed with aqueous sodium bicarbonate, extracted with DCM, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by column chromatography (PE/ea=5/1) to give compound 2 (2.0 g, yield 87%) as a yellow solid. MS m/z (ESI): 397.1 [ M+1].

A mixture of Compound 2 (2.0 g,7.9 mmol,1.0 eq), tert-butyl-3-aminoazetidine-1-carboxylic acid ester (2.1 g,5.1 mmol,1.5 eq) and cesium carbonate (4.9 g,15.2 mmol,3.0 eq) in DMF (100 mL) was stirred at 90℃under nitrogen for 30 min. The mixture was cooled to room temperature, extracted with DCM, washed with brine, dried over anhydrous sodium sulfate and concentrated, and the resulting residue was purified by flash column chromatography (PE/ea=4/1) to give compound 3 (2.4 g, 86% yield) as a yellow solid. MS m/z (ESI): 552.2 [ M+1].

To a solution mixture of compound 3 (2.4 g,4.4 mmol,1.0 eq) in dioxane (100 mL) at 0deg.C was added H ₂ SO ₄ (2.5 mL,4.4 mmol,1.0 eq) the mixture was stirred at room temperature 2 h. Adding NaOH aqueous solution (1.0M) to the mixture until the pH value is 8-9, extracting with ethyl acetate, washing with brine, and drying with anhydrous sodium sulfateDrying and concentration gave compound 4 (1.6 g, 80% yield) as a yellow solid. MS m/z (ESI): 452.2 [ M+1]]。

A solution of compound 4 (1.6 g,3.5 mmol,1 eq), (2-bromoethyl) 4, 5-pentafluoropentyl) sulfide (1.3 g,4.3 mmol,1.2 eq) and potassium carbonate (966 mg,7.0 mmol,2.0 eq) in acetonitrile (100 mL) was stirred at room temperature overnight. The mixture was concentrated and the resulting residue was purified by flash column chromatography (DCM/meoh=10/1) to give compound 5 (960 mg, 40% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.59 (s, 1H), 7.40 (d, J = 7.6 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 7.04-6.94 (m, 2H), 6.68 (d, J = 10.4 Hz, 2H), 5.24 (s, 1H), 4.46-4.58 (m, 2H), 4.24-4.17 (m, 2H), 3.66-3.58 (m, 2H), 3.41-2.51 (m, 8H), 2.46-2.36 (m, 2H),2..01-1.89 (m, 2H), 1.31-1.20 (m, 2H), 1.06 (d, J = 6.8 Hz, 3H). MS m/z（ESI）：672.2 [M+1]。

To a mixture of compound 5 (960 mg,1.4 mmol,1.0 eq) and ethyl acetate/acetic acid (100 mL, v/v 15/1) was added hydrogen peroxide (476 mg,14 mmol,10 eq,30%), and the mixture was stirred at room temperature 4 h. The mixture was washed with aqueous sodium bicarbonate and extracted with ethyl acetate. The combined organic layers were dried and concentrated. The resulting residue was purified by column chromatography (DCM/meoh=10/1) to give compound I-1 (100 mg, yield 10%) as a white solid. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.60 (s, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 7.04-6.94 (m, 2H), 6.68 (d, J = 10.4 Hz, 2H), 5.24 (s, 1H), 4.46-4.58 (m, 2H), 4.24-4.17 (m, 2H), 3.66-3.58 (m, 2H), 3.42-2.52 (m, 8H), 2.46-2.36 (m, 2H),2..01-1.89 (m, 2H), 1.31-1.21 (m, 2H), 1.08 (d, J = 6.8 Hz, 3H). MS m/z（ESI）：688.2 [M+1]。

Test example 1 degradation of ERalpha protein by Compound I-1

1. Purpose of test

The degradation of ERalpha protein by Compound I-1 was determined according to DC ₅₀ And maximum degradation efficiency E _max The compounds were evaluated for their in vitro degradation activity.

2. Test method

MCF-7 cells (ATCC, HTB-22) were cultured in 88% RPMI 1640 (Invitrogen, 22400-089) medium containing 10% FBS (Excell Bio, FSP 500), 1% P/S (Hyclone, SV 30010), 1% Gl μtaMax (Invitrogen, 25030-061). On the first day of the experiment, after centrifugation, digestion and cell collection, the cell suspension concentration was adjusted to 8.75X10 ⁴ mu.L of the suspension was added to each well of a 384 well plate (Greiner, 781090) per mL, and the mixture was placed at 37℃in 5% CO ₂ The incubator was left overnight. The next day, compounds were formulated in 10 mM stock with 100% DMSO and diluted to 2.5 μm (5X) with 1640 complete medium followed by 4-fold concentration gradient dilutions for 10 concentration points. 1. Mu.M fulvestrant was used as Low Control (LC) and 0.5% DMSO was used as High Control (HC). 10. Mu.L of the corresponding compound was added to each well in 384 well plates at 37℃with 5% CO ₂ Incubate in incubator for 24 hours. On the third day, 50. Mu.L of 8% paraformaldehyde (EM Sciences, 15710-S) was added to each well of 384 well plates, and the mixture was fixed at room temperature for 40 minutes; mu.L of PBS was added to each well and washed 2 times, followed by 50. Mu.L of 0.1% Triton X-100 (Sigma, T9284) per well, and left at room temperature for 15 minutes; the plates were washed 5 times with PBS, then 50. Mu.L of blocking solution containing 0.1% Tween 20 (Sigma, P1379) was added to each well and left at room temperature for 1 hour; original liquid in the plate was aspirated, and primary antibody (Estrogen Receptor a (D8H 8) Rabbit mAb, cell Signal, 8644S) was used at 1: dilutions were made at 1000. Mu.L per well and incubated overnight at 4 ℃. On the fourth day, the plates were washed 5 times, then secondary antibodies (IRDye 800CW gold anti-Rabbit, licor,926-32211, 1:1000) were diluted with DRAQ5 (Cell Signal,4084L, 1:2000) with blocking solution and incubated for 1 hour at room temperature after addition of 25. Mu.L per well; the plate was then washed 5 times and signal values at 800 nm and 700nm were read in an Odyssey imaging system, respectively. The Inhibition was calculated by Ratio (800 nm/700 nm), i.e.% -Inhibition = (Assay well-average_hc)/(average_lc-average_hc) ×100%, DC was calculated in Prism 5 using "log (inhibitor) vs. response-Variable slope" fitting dose-effect curve ₅₀ The method comprises the steps of carrying out a first treatment on the surface of the Maximum degradation rate E of compound _max Levels of residual ERα in cells after 500 nM treatmentPercent of erα levels after treatment with 1000 nM fulvestrant.

3. Test results

Degradation activity of compounds on ERα was determined by In-Cell-Western assay, DC ₅₀ E and E _max See table 1.

Degradation of ERalpha protein by the compounds of Table 1

* Namely, CN201580069484.8 discloses a compound 277

Conclusion: the compound I-1 of the present invention has better degradation activity on ERalpha than the compound 277 disclosed by GDC-9545 and CN 201580069484.8.

Test example 2 inhibition of MCF-7 cell proliferation by Compound I-1

1. Purpose of experiment

Determination of the inhibitory effect of Compound I-1 on the proliferation of MCF-7 cells in vitro, according to IC ₅₀ The activity of the compounds was evaluated.

2. Experimental method

MCF-7 cells (ATCC, HTB-22) were cultured in medium (Gibco, 22400089) containing 10% fetal bovine serum (Hyclone, SV 30087.03), 1% PS (penicillin-streptomycin diab, gibco, 15070063) and 1% Glμtamax (Gibco, 35050061) 1640. On the first day of experiment, after centrifugation, digestion and cell collection, the cell concentration was adjusted with complete medium, inoculated into 384 well plates (Greiner-781091) at a cell density of 600 cells/well, 40. Mu.L of cell suspension per well, and the plates were placed at 37℃with 5% CO ₂ The incubator was left overnight. The next day, compounds were formulated in 10 mM stock with 100% DMSO and further diluted to 5 μm (10X) with 1640 medium followed by 4-fold concentration gradient dilutions for 8 concentration points. 2. Mu.M fulvestrant was used as Low Control (LC) and 0.5% DMSO was used as High Control (HC). 10. Mu.L of the corresponding compound was added to each well in 384 well plates at 37℃with 5% CO ₂ Culturing in an incubator for 6 days. On day eight, 25. Mu.LCellTiter-Glo (Promega, G7573) was added to each well of the plate and incubated at 400 rpm for 10 minutes at room temperatureAfter the clock, the luminescence signal is read in Envision (PerkinElmer). The Inhibition was calculated by the following formula,% Inhibition = (Assay well-average_hc)/(average_lc-average_hc) ×100%, using Prism 5"log (inhibitor) vs. response-Variable slope" fitting the dose-effect curve, calculating IC ₅₀ 。

3. Measurement results

Proliferation inhibition of MCF-7 cells by Compounds IC obtained by measuring ATP content in living cells by CellTiter-Glo ₅₀ See table 2.

Inhibition of MCF-7 cell proliferation by the compounds of Table 2

* Namely, CN201580069484.8 discloses a compound 277

Conclusion: compared with the compound 277 disclosed by GDC-9545 and CN201580069484.8, the compound I-1 disclosed by the invention has better inhibition effect on the proliferation of MCF-7 cells.

Test example 3 evaluation of permeability and transporter substrates

And (3) cells: caco-2 cells

Instrument: a pure water meter, ELGA LabWate; a biosafety cabinet, nuaine; constant temperature CO ₂ Incubator, thermo; a microplate reader, perkinElmer; LC-MS/MS, AB SCIEX.

The method comprises the following steps: the aim of this study was to determine the bi-directional permeability of compounds using the Caco-2 monolayer cell model and to assess whether they were transported by the P-glycoprotein (P-gp) efflux. Caco-2 cells were seeded into 96-well cell plates for the transfer experiments after 24 days of continuous culture. The compound was administered bi-directionally with or without verapamil at a concentration of 2.00 μm. After 120 minutes of incubation, samples of the top end A and the base end B were collected, and the content of the test sample in each sample was detected by a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Apparent permeability coefficient and efflux rate were calculated. The data are summarized in table 3.

TABLE 3 permeability test results

* Namely, CN201580069484.8 discloses a compound 277

Conclusion: the compound I-1 disclosed by the invention shows moderate permeability in Caco-2 cells, has lower efflux rate and is obviously superior to the compound 277 disclosed by GDC-9545 and CN 201580069484.8.

Claims (7)

1. A compound of formula I, pharmaceutically acceptable salts thereof,

。

2. the compound according to claim 1, wherein the compound has a structure represented by formula I-1,

。

3. a process for the preparation of a compound according to claim 2, or a pharmaceutically acceptable salt thereof, comprising the steps of:

。

4. a pharmaceutical composition comprising a compound of any one of claims 1-2, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. Use of a compound according to any one of claims 1-2, a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 4 for the manufacture of a medicament for the treatment of breast cancer.

6. The use of claim 5, wherein the breast cancer is an estrogen receptor positive breast cancer.

7. The use according to any one of claims 5 or 6, further comprising an additional drug selected from the group consisting of tasilib, paclitaxel, anastrozole, exemestane, cyclophosphamide, epirubicin, fulvestrant, letrozole, gemcitabine, trastuzumab, polyethylene glycol fegrastim, tamoxifen, docetaxel, toremifene, vinorelbine, capecitabine, or ixabepilone.