CN116217611B

CN116217611B - Cyclobutanone derivative, preparation method and application

Info

Publication number: CN116217611B
Application number: CN202310518929.2A
Authority: CN
Inventors: 刘荣; 刘波; 王校; 李伟; 丁刚; 江爱民
Original assignee: Chengdu Mahadalong Pharmaceutical Technology Co ltd
Current assignee: Chengdu Mahadalong Pharmaceutical Technology Co ltd
Priority date: 2023-05-10
Filing date: 2023-05-10
Publication date: 2023-07-11
Anticipated expiration: 2043-05-10
Also published as: CN116217611A

Abstract

The invention discloses a cyclobutanone derivative, a preparation method and application thereof, belonging to the field of pharmaceutical chemistry, wherein the cyclobutanone derivative is a compound shown in a formula Ia or Ib, and the compound shown in the formula Ia or Ib has good TRPV1 inhibition activity and anti-tumor activity.

Description

Cyclobutanone derivative, preparation method and application

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a cyclobutanone derivative, a preparation method and application thereof.

Background

Opioid drugs are the primary choice for the treatment of pathological pain, but their clinical applications are greatly limited by a variety of adverse reactions such as tolerance, addiction, constipation, respiratory depression, etc. Development of novel high-efficiency analgesic low-adverse reaction analgesic drugs is one of the largest clinical demands. TRPV1 (transient receptor potential vanilloid subfamily, transient receptor potential vanillic acid subtype 1) is used as a non-selective cation ligand gating channel, is mainly expressed in medium-and small-sized nociceptive sensory neurons and is sensitive to environmental temperature change, when the environmental temperature is more than or equal to 43 ℃, the TRPV1 channel is opened, a thermal stimulus signal is converted into an electric signal, an action potential is generated, and pain sense is conducted, so that TRPV1 mediates the action potential of pain receptors, and the pain sense is involved in pain modulation. Thus, TRPV1 channels play an important role in the generation and development of pathological pain, and are expected to be new targets for the treatment of pathological pain. In 2004, team Giovanni Appendino and team Vincenzo Di Marzo cooperated findings (Eur. J. Org. Chem.2004, 3413), PPAHV (phorbol 12-phenyl-acetate 13-acetate 20-homovanillate, a phorbol derivative) was derived to obtain phorbol derivatives with cyclobutanone and cyclobutanol structures, most of which showed good activity on TRPV1 (its EC50 value range was around 10. Mu.M).

Primary alcohols and derivatives thereof, such as EBC-46 (tigilanol tiglate), can activate some forms of PKC (protein kinase C), promote a local immune response against the tumor, which in turn destroys the blood vessels of the tumor, and eventually kills cancer cells.

Therefore, the production cost is high in view of the complex structure of primary Buddha alcohols. The invention aims to obtain the cyclobutanone derivative which has the same biological activity, is simple in structure and easy to synthesize and manufacture, and the small molecular compound of the derivative has good TRPV1 inhibition activity and anti-tumor activity.

Disclosure of Invention

The invention provides a cyclobutanone TRPV1 inhibitor which has an anti-tumor activity effect. Specifically provides a cyclobutanone derivative with TRPV1 inhibition activity and anti-tumor activity, a preparation method and application thereof, wherein the derivative is a compound shown in a formula Ia or a formula Ib.

To achieve the object of the present invention, the following embodiments are provided.

In one embodiment selected from the group consisting of compounds of formula Ia or formula Ib,

，

in the method, in the process of the invention,

R ₁ is a substituted or unsubstituted siloxyl group;

R ₂ is H;

R ₃ is H, C C4 alkyl, propenyl or-CH 2C (O) CH3 group.

Preferably, the method comprises the steps of,

R ₁ is that

；

R ₂ Is H;

R ₃ is H, methyl, ethyl, -CH ₂ -CH=CH ₂ Radical or-CH ₂ C(O)CH ₃ A base.

In a specific embodiment, the compounds of formula Ia or formula Ib of the present invention are selected from:

。

the compounds of formula Ia or formula Ib according to the invention can be prepared by the following reaction scheme:

，

in the above-mentioned reaction scheme, the reaction mixture,

R ₁ is a substituted or unsubstituted siloxyl group;

R ₂ is H;

R ₃ is H, C C4 alkyl, propenyl or-CH 2C (O) CH3 group.

Preferably, the method comprises the steps of,

R ₁ is that

；

R ₂ Is H;

R ₃ is H, methyl, ethyl, -CH ₂ -CH=CH ₂ A group or a-CH 2C (O) CH3 group.

Further converting the compound of formula Ia into its derivative compound of formula Ib.

The invention also provides application of the compound shown in the formula Ia or the formula Ib or pharmaceutically acceptable salt in preparing a medicament for treating tumors. The tumor is selected from cervical cancer, bladder cancer, lung cancer, colon cancer, liver cancer and osteosarcoma.

The invention also provides a pharmaceutical composition containing the compound shown in the formula Ia or the formula Ib or pharmaceutically acceptable salt and pharmaceutical excipients.

The administration mode of the pharmaceutical composition is oral solid preparations such as tablets and capsules, injections, granules and oral liquid.

The auxiliary materials comprise filling agents, disintegrating agents, binding agents, lubricating agents, antioxidants, preservatives, sweeteners and the like, and can be prepared from auxiliary materials and processes conventional in the art.

Detailed Description

The following detailed description of the invention is provided by way of specific examples to aid in understanding the spirit of the invention and is not intended to limit the true scope of the invention in any way.

EXAMPLE 1 Synthesis of Compound 1

，

The first step: cuI (1.91 g,10 mmol) and Et ₂ O (diethyl ether, 200 mL) was added to a 500mL round bottom flask, the round bottom flask was placed in a-78deg.C low temperature bath and stirred for 30min, then compound 1-1 (0.5M in THF,100mL,50mmol) was added dropwise to the round bottom flask and stirred for 30min, and then the round bottom flask was placed in a-30deg.C low temperature bath and stirred for 30min; then, compound 1-2 (3.91 mL,50 mmol) was added dropwise to a round bottom flask via syringe over 20min, and reacted at-30℃for 30min; 200mL of saturated ammonium chloride solution was added dropwise to a round bottom bottle to quench the reaction and allowed to warm to room temperature, the aqueous phase was quenched with Et ₂ O (diethyl ether, 100 mL. Times.3) extraction, drying the combined organic phases over anhydrous sodium sulfate, suction filtration, and rotary evaporation concentration to constant weight at 5℃afforded compound 1-3 (5.92 g, yield=80%) as a colourless oil.

And a second step of: compounds 1-3 (5.92 g,40 mmol), TBDPSCl (t-butyldiphenylchlorosilane, 13.19g,48.0 mmol) DMF (N, N-dimethylformamide, 100 mL) and imidazole (6.81 g,100 mmol) were added to a 250mL round bottom bottle, the round bottom bottle was placed in an oil bath at 60℃and stirred for 2h, cooled to room temperature, EA (ethyl acetate, 300 mL) was added to the round bottom bottle, the organic phase was washed with water (100 mL. Times.3), the organic phase was dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and chromatographed on column (PE to PE/EA=200/1 as eluent) to give compounds 1-4 as colorless liquids (11.62 g, yield=75%).

And a third step of: compounds 1-4 (11.62 g,30 mmol), naI (sodium iodide, 13.19g,48.0 mmol) and acetone (60 mL) were added to a 250mL tube seal, the tube seal was placed in a 100deg.C oil bath and stirred for 24h, cooled to room temperature, 200mL of water was added after rotary evaporation of the acetone was concentrated off, the aqueous phase was extracted with PE (200 mL 3 times), the organic phases were combined and dried over anhydrous sodium sulfate, suction filtered, rotary evaporation was concentrated and column chromatographed (PE as eluent) to give compound 1 as a colourless oil (13.64 g, yield=95%). ¹ H-NMR(400MHz,CDCl ₃ )δ7.73–7.66(m,4H)，7.46–7.35(m,6H)，4.98–4.90(m,1H)，3.41(dtd,J=6.9,5.5,4.3Hz,1H)，3.14(d,J=4.1Hz,2H)，2.33–2.17(m,2H)，1.63(d,J=0.9Hz,3H)，1.49(s,3H)，1.08(s,9H)。

EXAMPLE 2 Synthesis of Compound 2

，

The first step: naHMDS (2.0M in Toluene,3.1mL) and THF (tetrahydrofuran, 10 mL) were added to a 50mL long neck single port flask, the reaction flask was placed in a-78 ℃ low temperature bath and stirred for 30min, then compound 2-1 (679.1 mg,6 mmol) was added dropwise to a round bottom flask and stirred for 30min; the reaction flask was placed in a low temperature bath at 0 ℃ and stirred for 30min, then compound 1 (3.01 g,6.3 mmol) was added, then the reaction flask was placed in an oil bath at 60 ℃ and stirred for 8h; after cooling to room temperature, the reaction was quenched by addition of 10mL of saturated aqueous ammonium chloride, extracted with EA (ethyl acetate, 10ml×3 times), the combined organic phases were dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and chromatographed on a column (PE/ea=5/1 as eluent) to give compound 2-2 as a colorless oily liquid (2.32 g, yield=83%).

And a second step of: compound 2-2 (2.32g,5 mmol), 2, 6-di-tert-butyl-4-methylpyridine (1.64 g,1.6eq.,8 mmol) and 1,2-DCE (1, 2-dichloroethane, 40 mL) were added to a 250mL two-necked flask, the flask was placed in an oil bath at 100deg.C and Tf ₂ O (trifluoromethanesulfonic anhydride, 1.0mL,6 mmol) was dissolved in 1,2-DCE (10 mL) and added dropwise to the reaction flask over 12h with syringe pump, cooled to room temperature, the reaction solution was concentrated and K was added sequentially ₂ CO ₃ (1.0g)、H ₂ O (50 mL) and THF (50 mL), the reaction was warmed to 65℃and stirred for 4h, then cooled to room temperature. The reaction mixture was concentrated, then 1N aqueous HCl (80 mL) was added, suction filtered and washed with t-BuOMe (50 mL. Times.4), and the aqueous phase was extracted with t-BuOMe (methyl tert-butyl ether, 50 mL. Times.4) after separation of the filtrate. The organic phases were combined and dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and purified by column chromatography (PE/etoac=20/1 to PE/etoac=10/1 as eluent), the product was concentrated and pumped down to constant weight with oil pump under reduced pressure to give compound 2 (749.89 mg, yield= 38%) as a yellowish oily substance. ¹ H-NMR(400MHz,CDCl ₃ )δ7.55(m,J=7.9,3.3,1.5Hz,4H)，7.36–7.27(m,6H)，3.91(t,J=4.8Hz,1H)，2.45(d,J=4.6Hz,2H)，2.37–2.26(m,2H)，2.11-1.89(m,2H)，0.98(s,9H)，0.93(s,6H)。

EXAMPLE 3 Synthesis of Compound 3

，

The first step: naHMDS (2.0M in Toluene,3.1mL) and THF (tetrahydrofuran, 10 mL) were added to a 50mL long neck single port flask, the flask was placed in a-78℃low temperature bath and stirred for 30min, then compound 3-1 (763, 2mg,6 mmol) was added dropwise to a round bottom flask and stirred for 30min; the reaction flask was placed in a low temperature bath at 0 ℃ and stirred for 30min, then compound 1 (3.05 g,6.3 mmol) was added, then the reaction flask was placed in an oil bath at 60 ℃ and stirred for 8h; after cooling to room temperature, the reaction was quenched with 10mL of saturated aqueous ammonium chloride, extracted with EA (ethyl acetate, 10ml×3 times), the combined organic phases were dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and chromatographed on a column (PE/ea=5/1 as eluent) to give compound 3-2 (2.26 g, yield=79%) as a colourless oil.

And a second step of: will be combined3-2 (2.26 g,5 mmol), 2, 6-di-tert-butyl-4-methylpyridine (1.64 g,1.6eq.,8 mmol) and 1,2-DCE (1, 2-dichloroethane, 40 mL) were added to a 250mL two-necked flask, the flask was placed in an oil bath at 100deg.C and Tf ₂ O (trifluoromethanesulfonic anhydride, 1.0mL,6 mmol) was dissolved in 1,2-DCE (10 mL) and added dropwise to the reaction flask over 12h with syringe pump, cooled to room temperature, the reaction solution was concentrated and K was added sequentially ₂ CO ₃ (1.0g)、H ₂ O (50 mL) and THF (50 mL), the reaction was warmed to 65℃and stirred for 4h, then cooled to room temperature. The reaction mixture was concentrated, then 1N aqueous HCl (80 mL) was added, suction filtered and washed with t-BuOMe (50 mL. Times.4), and the aqueous phase was extracted with t-BuOMe (methyl tert-butyl ether, 50 mL. Times.4) after separation of the filtrate. The organic phases were combined and dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and purified by column chromatography (PE/etoac=20/1 to PE/etoac=10/1 as eluent), and the product was concentrated and pumped down to constant weight with oil pump under reduced pressure to give compound 3 (616.8 mg, yield= 32%). ¹ H-NMR(400MHz,CDCl ₃ )δ7.55(ddd,J=7.9,3.3,1.5Hz,4H)，7.36–7.27(m,6H)，3.87(t,J=4.7Hz,1H)，2.42(d,J=4.6Hz,1H)，2.35–2.24(m,2H)，2.10(dd,J=14.5,4.5Hz,1H)，1.83(d,J=14.3Hz,1H)，0.97(s,9H)，0.91(s,6H)，0.79(s,3H)。

EXAMPLE 4 Synthesis of Compound 4

，

The first step: 3-ene-1-pentanoic acid (3.00 mL,36.0 mmol) and DCM (100 mL) were added to a 250mL reaction flask, HOBt (7.90 g,45.0 mmol) and EDCI HCl (8.70 g,45.0 mmol) were added, and after stirring at room temperature for 30min, cooled to 0deg.C, NEt was added ₃ (6.25 mL,45.0 mmol) and tetrahydropyrrole (3.0 mL,30.0 mmol) were stirred at room temperature overnight, the reaction mixture was concentrated and purified by column chromatography (PE/EtOAc=1/1 to EtOAc as eluent) and the product was concentrated and pumped down to constant weight by an oil pump to give compound 4-2 (4.13 g, yield=90%) as a pale yellow oil.

And a second step of: naHMDS (2.0M in Toluene,11mL) and THF (tetrahydrofuran, 200 mL) were added to a 500mL long neck single port flask, the flask was placed in a-78℃low temperature bath and stirred for 30min, then compound 4-2 (2.78 g,20 mmol) was added dropwise to a round bottom flask and stirred for 30min; the reaction flask was placed in a low temperature bath at 0 ℃ and stirred for 30min, then compound 1 (10.53 g,22 mmol) was added, then the reaction flask was placed in an oil bath at 60 ℃ and stirred for 8h; after cooling to room temperature, the reaction was quenched by addition of 200mL of saturated aqueous ammonium chloride, extracted with EA (ethyl acetate, 200ml×3 times), the combined organic phases were dried over anhydrous sodium sulfate, filtered off with suction, concentrated by rotary evaporation and chromatographed on a column (PE/ea=5/1 as eluent) to give compound 4-3 as a colorless oily liquid (9.0 g, yield=89%).

And a third step of: compounds 4-3 (9.0 g,17.9 mmol), 2,6-tBu-4-MePy (5.8 g,28.6 mmol) and DCE (160 mL) were added to a 500mL two-necked flask, and after the flask was placed in an oil bath at 100deg.C, tf was measured ₂ O (3.80 mL,22.3 mmol) was dissolved in DCE (20 mL) and added dropwise to the flask over 12h with syringe pump, cooled to room temperature, the reaction was concentrated and K was added sequentially ₂ CO ₃ (5.0g)、H ₂ O (100 mL) and THF (100 mL), the reaction was warmed to 65℃and stirred for 4h, then cooled to room temperature. The reaction mixture was concentrated, and then added with 1N HCl (80 mL),tBuOMe extraction (50 mL. Times.4). The organic phases were combined over anhydrous Na ₂ SO ₄ After drying, suction filtration and concentration, column chromatography separation and purification were performed (PE/etoac=20/1 to PE/etoac=10/1 as eluent), and the product was pumped down to constant weight by oil pump after concentration to give 2310.7mg (30% yield) of compound 4-4 as a colorless oil.

Fourth step: compound 4-4 (350 mg,0.80 mmol) was dissolved in THF (5 mL) and added to a 25mL reaction flask and cooled to-25deg.C, 2-bromopropylmagnesium bromide (1M in THF,2.40mL,3.0eq.,2.40 mmol) was added and the reaction was carried out overnight at-25deg.C, saturated NH was added ₄ The reaction was quenched with Cl solution (1.0 mL) and then H was added ₂ O (5 mL) was diluted and EtOAc extracted (5 mL. Times.3). The organic phases were combined over anhydrous Na ₂ SO ₄ Drying, suction filtering and concentrating, separating and purifying by column chromatography (PE is eluent), concentrating, pumping under reduced pressure to constant weight by oil pump, and obtaining colorless oily liquid 200.4mg. Weighing Pd (OAc) in a glove box ₂ (95.0 mg,1.0eq.,0.42 mmol) in a 25mL reaction flask, 200.4mg of the resulting viscous liquid (0.42 mmol,1.0 eq.)Dissolve in DCE (5 mL) and add to the system and stir at room temperature for 10h. After the reaction solution was concentrated, column chromatography was performed for separation and purification (PE/etoac=15/1 to PE/etoac=5/1 as eluent), and the product was pumped down to constant weight by an oil pump after concentration to give compound 4 (78.1 mg, yield=16%, single crystal cultivation with PE/etoac=20/1) as a white solid. ¹ H-NMR(400MHz,CDCl ₃ ):δ7.62(d,J=6.6Hz,4H)，7.40–7.31(m,6H)，4.52(s,1H)，4.50–4.44(m,1H)，4.18–4.09(m,1H)，2.88(d,J=15.7Hz,1H)，2.32(dd,J=14.2,6.3Hz,2H)，2.10(s,3H)，2.03(ddd,J=24.8,11.6,4.7Hz,2H)，1.96–1.91(m,2H)，1.53(s,3H)，1.43(d,J=4.3Hz,3H)，1.19(s,3H)，1.05(s,9H)。

EXAMPLE 5 TRPV1 Activity inhibition assay

The test uses human embryo kidney 293 cells stably transfected with TRPV1 as a detection object, wherein the TRPV1-HEK293 cells are firstly incubated with Ca < 2+ > dye, and basic fluorescence signals F0 are collected at sampling intervals of 1s within the range of 515-575 nm under the excitation wavelength of 470-495 nm. After the start of the test, the baseline fluorescence intensity of the cell plate was continuously measured for 60s without adding a sample, and the average value of the fluorescence signals at the first 10 time points was used as the base fluorescence intensity value (F0). Subsequently, the sample was automatically applied and the fluorescence intensity value (F) was continuously measured for 400s for 460s. The average value F and the relative fluorescence intensity F/F0 of the fluorescence intensity F of each well at each test time point are calculated respectively, and a time-F/F curve is generated. Calculating the Area Under the Curve (AUC) of the time-F/F0 Curve from 60s to 300s, taking the AUC value as a measure of the calcium ion influx effect, and calculating the EC according to the AUC value ₅₀ The results are shown in Table 1.

TABLE 1 inhibition of TRPV1 Activity of Compounds 1-4 and 4-4

The above bioactivity test data shows that: compounds 2, 3, 4 have good inhibition of TRPV1 activity.

EXAMPLE 6 anti-tumor biological Activity test

Culturing Hela (cervical cancer cells), T24 (bladder cancer cells), A549 (lung cancer cells), hepG2 (liver cancer cells), HCT-8 (colon cancer cells), U2OS (osteosarcoma cells) and the like according to standard methods, incubating for 24 hours at 24 ℃ in an incubator according to ATCC standard methods, adding the compound (DMSO solution) prepared in the above examples 1-4, incubating for 24 hours in the incubator, measuring absorbance (a) value at a wavelength of 450nm by using a cck8 method, and calculating the inhibition effect of the compound on the above cells. The results are shown in Table 2.

TABLE 2 tumor cell Activity of Compounds 1-4 and 4-4

The above bioactivity test data shows that: the compound 2 has excellent inhibitory activity on cervical cancer cells, bladder cancer cells, liver cancer cells and colon cancer cells; the compound 3 has excellent inhibitory activity on cervical cancer cells, bladder cancer cells, liver cancer cells, colon cancer cells and lung cancer cells; the compound 4 has excellent inhibitory activity on cervical cancer cells, bladder cancer cells, lung cancer cells and liver cancer cells, and the compound 4-4 has excellent inhibitory activity on cervical cancer cells and bladder cancer cells.

The foregoing is a typical example of the invention and any simple modification or variation made within the spirit of the invention is also within the scope of the invention.

Claims

1. A cyclobutanone derivative, characterized in that: is a compound shown in a formula Ia or a pharmaceutically acceptable salt thereof,

in the method, in the process of the invention,

R ₁ is that

R ₂ Is H;

R ₃ is H, C ₁ -C ₄ Alkyl, allyl or-CH ₂ C(O)CH ₃ A base.

2. The cyclobutanone derivative of claim 1, wherein said R ₃ Is H, methyl, ethyl, -CH ₂ -CH＝CH ₂ or-CH ₂ C(O)CH ₃ A base.

3. The cyclobutanone derivative of claim 1, wherein said compound of formula Ia or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

4. a cyclobutanone derivative, which is a compound shown in a formula 4 or a pharmaceutically acceptable salt thereof,

5. a process for the preparation of a compound of formula Ia as claimed in any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, comprising:

1) Reacting a compound of formula IV with a compound of formula III to obtain a compound of formula II;

2) Cyclizing the compound of formula II to obtain the compound of formula Ia.

6. Use of a compound of formula Ia as described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a tumour.

7. The use according to claim 6, wherein the tumor is selected from cervical cancer, bladder cancer, lung cancer, colon cancer, liver cancer and osteosarcoma.

8. A pharmaceutical composition comprising a compound of formula Ia or formula 4 as defined in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant.