CN109593061A - Androgen receptor antagonists, preparation method and its application - Google Patents

Abstract

The present invention relates to a kind of androgen receptor antagonists, preparation method and its application, which is the compound that general formula (1) indicates and its pharmaceutically acceptable salt (R in general formula₁、R₂、R₃And R₄Definition as shown in specification).

Description

Androgen receptor antagonists, preparation method and its application

Technical field

The present invention relates to pharmaceutical technology fields, in particular to a kind of novel androgen receptor antagonists, its preparation side Method and its application.

Background technique

Prostate-cancer incidence occupies second in all malignant tumours of male.Androgen receptor (Androgen Receptor, AR) it is the important target for treating prostate cancer, the combination of androgen and AR is blocked using androgen receptor antagonists Therapeutic purposes can be reached.Clinically used nonsteroidal androgens receptor antagonist have Bicalutamide (R-bicalutamide) and The miscellaneous Shandong amine (Enzalutamide) of grace, structure is as follows.

These compounds are used to treatment prostate cancer, and Bicalutamide (US4636505) is Astrazeneca AB's exploitation, Nineteen ninety-five listing, trade name Kang Shi get (Casodex).Miscellaneous Shandong amine (MDV3100, the US2007254933) trade name of grace Xtandi is developed jointly by Medivation company and Astellas company, is supported in August, 2012 through FDA approval treatment trend Refractory prostate cancer.Marcella Bassetto, Salvatore Ferla etc. is in " Design and synthesis of novel bicalutamide and enzalutamide derivatives as antiproliferative agents for the treatment of prostatecancer(European Journal of Medicinal Chemistry 118 (2016) 230-243) " in studies have shown that Bicalutamide is living in 22Rv1, DU-145, LNCaP and VCaP cell strain anticancer Property IC50 geometric mean be 52.42 μM, the miscellaneous Shandong amine activity geometric mean of grace is then 28.10 μM, their bioactivity It is universal not high, to reach therapeutic purposes, larger dose is needed to carry out using to increase patient's metabolic burden.In addition, than card Shandong There are also the effects (agonist) for activating androgen receptor for amine, to stimulate cancer hyperplasia (CN104024228B).

Summary of the invention

The present invention is completed in view of above-mentioned project existing in the prior art, and its purpose is to provide a kind of novel The androgen receptor antagonists of high-efficiency low-toxicity, pharmaceutical activity is high, and dosage is low, and toxic side effect is small.

The present invention provides the compound and its pharmaceutically acceptable salt of the following general formula (1) expression,

Wherein, R₁With R₂It is identical or different, respectively indicate hydrogen atom or R₁With R₂It is formed together C_3-6Naphthenic base；

R₃Indicate hydrogen atom or C_1-8Alkyl；

R₄For substituted or unsubstituted C_6-10Aryl or heteroaryl, the heteroaryl are 5- or 6-membered monocycle base or are Bicyclic group made of 5- or 6-membered monocycle and phenyl ring condensation.

In a preferred embodiment, R₄For the C at least one substituent group_6-10Aryl or heteroaryl, the substituent group are selected from Halogen atom, cyano, nitro, C_1-8Alkyl, C_1-8Alkoxy, halogenated C_1-8Alkyl, halogenated C_1-8Alkoxy, halogenated C_1-8Alkylthio group, Halogenated C_1-8Alkyl sulphinyl, halogenated C_1-8Alkyl sulphonyl or pentafluoride-sulfanyl.

In another preferred embodiment, R₄For selected from any one of following radicals:

In formula, n indicates 1~7 integer.

In addition, the present invention provides a kind of pharmaceutical composition, containing compound of any of claims 1-3 or Its pharmaceutically acceptable salt and pharmaceutically acceptable carrier.

In addition, the invention further relates to above compound or its pharmaceutically acceptable salt preparation for prevent or treat with Application in the drug of the relevant disease of estrogen receptor activity.Wherein, the disease packet relevant to estrogen receptor activity It includes: hormone-sensitive prostate cancer, hormone-refractory prostate cancer, benign prostatic hyperplasis, acne, hirsutism, whelk, dark Sore and alopecia.

In addition, a kind of preparation method of the compound indicated the present invention also provides general formula (1), includes the following steps:

Step (I) reacts compound (2) with oxalyl chloride or thionyl chloride in aprotic solvent, forms compound (3)；

Step (II) reacts compound (3) with compound (4) in aprotic solvent, is formed compound (5)；

Step (III) in organic solvent reacts compound (5) with piperidines, is formed compound (6)；

Step (IV) in organic solvent reacts compound (6) with compound (7), is formed compound (1)；

R in the above formulas₁、R₂、R₃And R₄Definition it is same as above.

Invention effect

(1) the compound of the present invention is the androgen antagonist of structure novel and excellent effect, can be used for treating androgen Related disease has stronger inhibiting effect to prostate gland cancer cell, and cell activity is that clinical application Bicalutamide and grace are miscellaneous The several times of Shandong amine.

(2) preparation method of the compound of the present invention is simple.

Specific embodiment

R used in this paper general formula₁、R₂、R₃And R₄The substituent group of the group of expression and these groups is as described below.

“C_1-8Alkyl " refers to straight chain-or branched-Alkyl with 1 to 8 carbon atom, for example, methyl, second Base, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl, sec-butyl, n-pentyl, neopentyl, n-hexyl, isohesyl, 3- first Base amyl, heptyl and octyl etc..

“C_1~8Alkoxy " refers to straight chain-or branch-alkoxy (C with 1 to 8 carbon atom_1-8Alkyl-O-), such as Can enumerate methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, N-pentyloxy, neopentyl oxygen, positive hexyloxy, dissident's oxygroup, 3- methyl amoxy, positive oxygroup in heptan, n-octyloxy etc..

“C_1~8Alkylthio group " refers to straight chain-or branch-alkylthio group (C with 1 to 8 carbon atom_1-8Alkyl-S-), such as Can enumerate methyl mercapto, ethylmercapto group, positive rosickyite base, isopropyisulfanyl, positive butylthio, isobutylthio, tertiary butylthio, secondary butylthio, Positive penta sulfenyl, new penta sulfenyl, just own sulfenyl, isohexylthio, 3- methyl penta sulfenyl, positive sulfenyl in heptan, just pungent sulfenyl etc..

" halogen atom " indicates fluorine, chlorine, bromine and iodine.

" halogenated C_1~8Alkyl " refers to above-mentioned " C_1~8Hydrogen atom in alkyl " is replaced and formed by least one halogen atom Group.Such as trihalomethyl group (such as-CF₃), three halogenated ethyls are (such as-CH₂CF₃), five halogenated ethyls are (such as-CF₂CF₃) or nine halogen For butyl (such as-CF₂CF₂CF₂CF₃) etc..

“C_3~6Naphthenic base " refers to 3~6 yuan of saturation hydrocarbon rings, for example, cyclopropyl, cyclobutyl, cyclopenta, ring Hexyl.

“C_6-10Aryl " refers to the aromatic hydrocarbyl with 6~10 carbon atoms, for example, phenyl, Alpha-Naphthyl, Betanaphthyl etc..

" heteroaryl " refers to 5- or 6-membered monocycle base, or is bicyclic made of above-mentioned 5- or 6-membered monocycle is condensed with phenyl ring Base.5- or 6-membered monocycle base be containing selected from heteroatomic 5~6 unit's heteroaryl of at least one of N, O, S, for example, Imidazoles, pyrazoles, pyridine, pyrazine, pyrimidine, pyridazine, thiophene, furans, pyrans, two pyrans, thiazole, isothiazole, thiadiazoles, thiazine, Oxazole, isoxazole, pyrroles, dioxazole etc..For bicyclic group made of 5- or 6-membered monocycle and phenyl ring condensation, for example, Benzimidazole, benzo pyridine, benzo pyrimidine, benzothiophene, benzofuran, benzothiazole, benzoxazoles, benzo isoxazole etc..

In addition, pharmaceutical composition of the invention, contains above-mentioned general formula (1) compound represented or its medicine as effective component Acceptable salt on.The pharmaceutical compositions include compound described in any one of above embodiment and variation as work Property ingredient, with pharmaceutically acceptable carrier, diluent or excipient composition.

Term " pharmaceutically acceptable salt " refers to the salt of the biological efficacy and property that keep the compounds of this invention, the salt one As in terms of biology or other aspects do not have detrimental effect.In many cases, the compound of the present invention can utilize presence Amino and/or carboxyl or similar group form hydrochlorate and/or alkali salt.

The compound of the present invention can be used in preventing or treat disease relevant to estrogen receptor activity, for example, hormone Sensitive prostate cancer, hormone-refractory prostate cancer, benign prostatic hyperplasis, acne, hirsutism, whelk, dark sore and de- Hair.

The compound of the present invention can also be prepared other than preparation method described above by following methods.

R in formula₁、R₂、R₃And R₄Definition it is same as above, R₅For tert-butyl or methyl.

Hereinafter, being illustrated by preparation method of the specific embodiment to the compound of the present invention.

Embodiment 1 (preparation of compound JD1001-2053)

5.0g (15.4mmol, 1.0eq) Fmoc-1- amino cyclopropane is added under nitrogen protection into 200mL there-necked flask Then acid sequentially adds DCM 50mL, 9.8g oxalyl chloride (76.9mmol, 5.0eq) is added dropwise in 4 drop DMF at room temperature, It is stirred to react at room temperature 4 hours, taking out two drop reaction solutions and methyl alcohol process is added is methyl esters, detects reaction by TLC and completes (expansion Agent: PE:EA=1:1), vacuum distillation removes oxalyl chloride and DCM, obtains faint yellow solid, inflated with nitrogen saves backup.

The faint yellow solid that previous step is reacted is taken, sequentially adds 90mL dry THF thereto, 1.9g (22.6mmol, 1.5eq)NaHCO₃, 3.4g (18.3mmol, 1.2eq) 5- amino -3- trifluoromethyl -2- cyanopyridine is heated to 60 DEG C, maintains Temperature is stirred to react 2 hours, is detected reaction by TLC and is completed (solvent: PE:EA=1:1), stops reacting and dropping to room temperature, 200mL water diluting reaction system is added, EA extracted products 100ml*3 times, organic phase is dried over anhydrous sodium sulfate, and is filtered, and is added 40mL piperidines is stirred to react 1 hour under room temperature, is detected reaction by TLC and is completed (solvent: PE:EA=1:1), 40 Vacuum distillation removes solvent under DEG C water bath condition, obtains 4 grams of solids.Column chromatography for separation is carried out using silica gel, eluant, eluent is petroleum Ether and ethyl acetate (PE:EA=5:1~1:1), finally obtain 0.93g solid chemical compound JD1001-002-9, yield 22%.

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 4- fluorophenylsulfonyl chloride 0.22g (1.11mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and is concentrated to give solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, desivac Obtain product J D1001-2053 about 28mg, yield 19%.

¹H NMR:(DMSO,ppm,400MHz)δ1.02(2H,m),1.33(2H,m),7.38(2H,m),7.85(2H,m), 8.57(2H,m),9.13(1H,d),10.60(1H,s).¹⁹F NMR:(DMSO, ppm, 400MHz) δ -61.34, -106.22. matter Spectrum: 429 [M+H]⁺.

Embodiment 2 (preparation of compound JD1001-2054)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 3,5- difluoro chloride 0.24g (1.11mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, Desivac obtains product J D1001-2054 about 24mg, yield 16%.

¹H NMR:(DMSO,ppm,400MHz)δ1.07(2H,m),1.40(2H,m),7.51(2H,m),7.57(1H,m), 8.60(1H,d),8.85(1H,s),9.18(1H,d),10.64(1H,s).¹⁹F-NMR:(DMSO,ppm,400MHz)δ- 61.33, -106.21. mass spectrum: 447 [M+H]⁺.

Embodiment 3 (preparation of compound JD1001-2055)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 3,4,5- trifluoro benzene sulfonyl chloride 0.26g (1.11mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, Desivac obtains product J D1001-2055 about 45mg, yield 30%.

¹H NMR:(DMSO,ppm,400MHz)δ1.08(2H,m),1.39(2H,m),7.73(2H,m),8.62(1H,d), 8.84(1H,d),9.21(1H,d),10.63(1H,s).¹⁹F NMR:(DMSO, ppm, 400MHz) δ -61.40, -131.20, - 131.25. mass spectrum: 465 [M+H]⁺.

Embodiment 4 (preparation of compound JD1001-2056)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 4- trifluoromethyl benzene sulfonyl chloride 0.27g (1.11mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, Desivac obtains product J D1001-2056 about 27mg, yield 24%.

¹H NMR:(DMSO,ppm,400MHz)δ1.17(2H,m),1.39(2H,m),7.86(2H,d),8.01(2H,d), 8.47(1H,d),8.75(1H,s),8.99(1H,d),10.48(1H,s).¹⁹F NMR:(DMSO,ppm,400MHz)δ- 61.51, -62.01. mass spectrum: 479 [M+H]⁺.

Embodiment 5 (preparation of compound JD1001-2057)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 6- trifluoromethyl pyridine -3- sulfonic acid chloride 0.27g (1.11mmol, 3.0eq) is stirred to react 3 hours under room temperature, by TLC detect reaction complete (solvent: PE:EA=1: 1) saturation NaHCO, is added₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, has with the washing of 0.5N aqueous hydrochloric acid solution Machine phase 2 times, anhydrous sodium sulfate dries, filters, and is concentrated to give solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) separates pure Change, desivac obtains product J D1001-2057 about 46mg, yield 32%.

¹H NMR:(DMSO,ppm,400MHz)δ1.20(2H,m),1.46(2H,m),8.06(1H,d),8.47(2H,m), 9.05(1H,d),9.11(1H,d),10.48(1H,s).¹⁹F NMR:(DMSO, ppm, 400MHz) δ -61.51, -67.15. matter Spectrum: 480 [M+H]⁺.

Embodiment 6 (preparation of compound JD1001-2060)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 2- trifluoromethyl benzene sulfonyl chloride 0.27g (1.11mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, Desivac obtains product J D1001-2060 about 39mg, yield 25%.

¹H NMR:(DMSO,ppm,400MHz)δ1.03(2H,m),1.40(2H,m),7.90(3H,m),8.12(1H,d), 8.57(1H,d),8.82(1H,s),9.13(1H,d),10.51(1H,s).¹⁹F NMR:(DMSO,ppm,400MHz)δ- 56.04, -61.28. mass spectrum: 479 [M+H]⁺.

Embodiment 7 (preparation of compound JD1001-2061)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-9 solid 0.10g (0.37mmol, 1.0eq), THF 3.2mL, DMAP 0.14g (1.11mmol, 3.0eq), 3,5- bis- (trifluoromethyl) benzene sulfonyl chloride 0.35g (1.11mmol, 3.0eq) is stirred to react 3 hours under room temperature, by TLC detect reaction complete (solvent: PE:EA=1: 1) saturation NaHCO, is added₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, has with the washing of 0.5N aqueous hydrochloric acid solution Machine phase 2 times, anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40) It isolates and purifies, desivac obtains product J D1001-2061 about 40mg, yield 36%.

¹H NMR:(DMSO,ppm,400MHz)δ1.22(2H,m),1.46(2H,m),8.32(2H,s),8.36(1H,s), 8.42(1H,d),9.01(2H,m),10.47(1H,s).¹⁹F NMR:(DMSO, ppm, 400MHz) δ -61.61, -61.68. matter Spectrum: 547 [M+H]⁺.

Embodiment 8 (preparation of compound JD1001-2063)

5.0g (14.8mmol, 1.0eq) Fmoc-1- amino cyclobutane carboxylic is added under nitrogen protection into 200mL there-necked flask Then acid sequentially adds DCM 50mL, 9.4g (74.0mmol, 5.0eq) oxalyl chloride is added dropwise in 4 drop DMF at room temperature, It is stirred to react at room temperature 4 hours, taking out two drop reaction solutions and methyl alcohol process is added is methyl esters, detects reaction by TLC and completes (expansion Agent: PE:EA=1:1), vacuum distillation removes excessive oxalul chloride and DCM, obtains faint yellow solid, inflated with nitrogen saves backup.

The faint yellow solid that previous step is reacted is taken, sequentially adds 90mL dry THF thereto, 1.9g (22.2mmol, 1.5eq)NaHCO₃, 3.3g (17.8mmol, 1.2eq) 5- amino -3- trifluoromethyl -2- cyanopyridine is heated to 60 DEG C, maintains Temperature is stirred to react 2 hours, is detected reaction by TLC and is completed (solvent: PE:EA=1:1), stops reacting and dropping to room temperature, 200mL water diluting reaction system, EA extracted products 3 times (100ml*3 times) is added, organic phase is dried over anhydrous sodium sulfate, it filters, 40mL piperidines is added, is stirred to react under room temperature 1 hour, reaction is detected by TLC and completes (solvent: PE:EA=1:1), Vacuum distillation removes solvent under 40 DEG C of water bath conditions, obtains 4 grams of solids.Column chromatography for separation is carried out using silica gel, eluant, eluent is Petroleum ether and ethyl acetate (PE:EA=5:1~1:1), finally obtain 1.0g solid chemical compound JD1001-002-10, yield 23%.

0.10g (0.35mmol, 1.0eq) JD1001-002-10 is added under the conditions of nitrogen protection into 25mL round-bottomed flask Solid, 3.2mL THF, 0.13g (1.05mmol, 3.0eq) DMAP, 0.20g (1.05mmol, 3.0eq) 4- fluorophenylsulfonyl chloride, room It is stirred to react under the conditions of temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), saturation NaHCO is added₃Water Organic phase is obtained by extraction in solution quenching reaction, ethyl acetate, with 0.5N aqueous hydrochloric acid solution washing organic phase 2 times, anhydrous sodium sulfate It dries, filters, concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, desivac obtains Product J D1001-2063 about 28mg, yield 21%.

¹H NMR:(DMSO,ppm,400MHz)δ1.60(1H,m),1.71(1H,m),2.13(2H,m),2.40(2H,m), 7.31(2H,t),7.78(2H,m),8.56(2H,m),9.10(1H,d),10.60(1H,s).¹⁹F NMR:(DMSO,ppm, 400MHz) δ -61.40, -106.58. mass spectrum: 443 [M+H]⁺.

Embodiment 9 (preparation of compound JD1001-2064)

0.10g (0.35mmol, 1.0eq) JD1001-002-10 is added under the conditions of nitrogen protection into 25mL round-bottomed flask Solid, 3.2mL THF, 0.13g (1.05mmol, 3.0eq) DMAP, 0.22g (1.05mmol, 3.0eq) 3,5- difluoro benzene sulfonyl Chlorine is stirred to react 3 hours under room temperature, is detected reaction by TLC and is completed (solvent: PE:EA=1:1), saturation is added NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, with 0.5N aqueous hydrochloric acid solution washing organic phase 2 times, nothing Aqueous sodium persulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, freezes Dry method obtains 16mg JD1001-2064 product, yield 9%.

¹H NMR:(DMSO,ppm,400MHz)δ1.65(1H,m),1.80(1H,m),2.18(2H,m),2.45(2H,m), 7.38(2H,m),7.46(1H,m),8.58(1H,d),8.98(1H,bs),9.10(1H,d),10.69(1H,s).¹⁹F NMR: (DMSO, ppm, 400MHz) δ -61.43, -106.77. mass spectrum: 461 [M+H]⁺.

Embodiment 10 (preparation of compound JD1001-2065)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-10 solid 0.10g (0.35mmol, 1.0eq), THF 3.2mL, DMAP 0.13g (1.05mmol, 3.0eq), 4- trifluoromethyl benzene sulfonyl chloride 0.26g (1.05mmol, 3.0eq), it is stirred to react under room temperature 3 hours, reaction is detected by TLC and completes (solvent: PE:EA=1:1), be added full And NaHCO₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, is washed organic phase 2 times with 0.5N aqueous hydrochloric acid solution, Anhydrous sodium sulfate dries, filters, and is concentrated to give solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40 it) isolates and purifies, desivac Obtain product J D1001-2065 about 39mg, yield 30%.

¹H NMR:(DMSO,ppm,400MHz)δ1.65(1H,m),1.81(1H,m),2.25(2H,m),2.47(2H,m), 7.78 (2H, d), 7.90 (2H, d), 8.45 (1H, d), 8.85 (1H, bs), 8.98 (1H, d), 10.55 (1H, bs) mass spectrums: 493 [M+H]⁺.

Embodiment 11 (preparation of compound JD1001-2066)

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-10 solid 0.10g (0.35mmol, 1.0eq), THF 3.2mL, DMAP 0.13g (1.05mmol, 3.0eq), 3,5- bis- (trifluoromethyl) benzene sulfonyl chloride 0.33g (1.05mmol, 3.0eq) is stirred to react 3 hours under room temperature, by TLC detect reaction complete (solvent: PE:EA=1: 1) saturation NaHCO, is added₃Organic phase is obtained by extraction in aqueous solution quenching reaction, ethyl acetate, has with the washing of 0.5N aqueous hydrochloric acid solution Machine phase 2 times, anhydrous sodium sulfate dries, filters, and concentration organic phase obtains solid, prepares liquid phase (CH through high pressure₃CN:H₂O=60:40) It isolates and purifies, desivac obtains product J D1001-2066 about 41mg, yield 34%.

¹H NMR:(DMSO,ppm,400MHz)δ1.69(1H,m),1.87(1H,m),2.27(2H,m),2.54(2H,m), 8.19 (2H, s), 8.29 (1H, s), 8.39 (1H, d), 8.95 (1H, d), 9.09 (1H, s), 10.53 (1H, s) mass spectrums: 561 [M +H]⁺.

Embodiment 12 (preparation of compound JD1001-2137)

1- aminocyclohexyl carboxylic acid methyl ester hydrochloride 3.9g is added under the conditions of nitrogen protection into 100mL round-bottomed flask (20.0mmol, 1.0eq) and THF 30mL, be successively added dropwise under the conditions of being stirred at room temperature triethylamine 3.0g (3.0mmol, 1.5eq), DMAP 244mg (2mmol, 0.1eq), 3,5- bis- (trifluoromethyl) benzene sulfonyl chloride 6.88g (22.2mmol, 1.1eq), Continue that reaction 3h is stirred at room temperature, reaction is detected by TLC and completes (solvent: PE:EA=1:1), saturation NaHCO is added₃It is water-soluble Liquid quenching reaction, EA extracted products 50ml*3 times, with 0.5N salt acid elution 2 times, anhydrous sodium sulfate dries, filters organic phase, dense Contract to obtain solid.

Previous step reaction solid is dissolved in 100mL ethyl alcohol, 30mL water is added, it is solid that 4g (5.0eq) sodium hydroxide is then added Body, 60 DEG C of oil bath heatings are stirred to react 2h, detect reaction by LCMS and complete, solvent is removed in vacuum distillation.DCM and H is added₂O Each 50ml stirs 0.5h, retains water phase after liquid separation, and 1N salt acid for adjusting pH value is added under the conditions of being stirred at room temperature to 5, there is a large amount of solids It is precipitated, after filtering drying, obtains 3.8 grams of faint yellow solids.Faint yellow solid 335mg (0.8mmol, 1.0eq) is added to In 25ml round-bottomed flask, 3mL DCM is sequentially added, 3 drip DMF, and oxalyl chloride 300mg is added dropwise under 0 DEG C of ice-water bath stirring condition (2.4mmol, 3.0eq) is then warmed to room temperature reaction 2 hours, and taking out two drop reaction solutions and methyl alcohol process is added is methyl esters, through LCMS Detection reaction is completed, and is concentrated under reduced pressure to give 350mg solid product JD1001-002-20, is saved backup under the conditions of nitrogen protection.

Under the conditions of nitrogen protection into 25mL round-bottomed flask be added JD1001-002-20 solid 109mg (0.25mmol, 1.0eq), THF 3.2mL, NaHCO₃63mg (0.75mmol, 3.0eq), 5- amino -3- trifluoromethyl -2- cyanopyridine 140mg (0.75mmol, 3.0eq), 60 DEG C of heating stirrings are reacted 3 hours, TLC detection reaction completion (solvent: PE:EA=1: 1) water 5ml quenching reaction, is added, organic phase is obtained by extraction in EA, after concentration, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies, Product J D1001-2137 about 18mg, yield 15% are obtained after freeze-drying.

¹H NMR:(DMSO,ppm,400MHz)δ1.1-1.3(6H,m),1.35-1.60(2H,m),1.8-2.0(2H,m), 8.24 (2H, s), 8.38 (1H, s), 8.44 (1H, s), 8.99 (1H, s), 10.25 (1H, s), 11.85 (1H, s) mass spectrums: 589 [M+H]⁺.

Embodiment 13 (preparation of compound JD1001-2139)

Aminoisobutyric acid methyl ester hydrochloride 3.1g is added under the conditions of nitrogen protection into 100mL round-bottomed flask (20.0mmol, 1.0eq) and 30mL THF, be successively added dropwise under the conditions of being stirred at room temperature triethylamine 3.0g (30.0mmol, 1.5eq), DMAP 244mg (2mmol, 0.1eq), 3,5- bis- (trifluoromethyl) benzene sulfonyl chloride 6.88g (22.2mmol, 1.1eq), Continue that reaction 3h is stirred at room temperature, reaction is detected by TLC and completes (solvent: PE:EA=1:1), saturation NaHCO is added₃It is water-soluble Liquid quenching reaction, EA extracted products 3 times (50ml*3 times), organic phase is with 0.5N salt acid elution 2 times, and anhydrous sodium sulfate is dry, mistake Filter, is concentrated to give solid.

Solid is dissolved in ethyl alcohol 100mL, water 30mL is added, sodium hydrate solid 4g (5.0eq) then is added, 60 DEG C of oil baths Heating stirring reacts 2h, detects reaction by LCMS and completes, solvent is removed in vacuum distillation.DCM and H is added₂Each 50ml stirring of O 0.5h retains water phase after liquid separation, and 1N salt acid for adjusting pH value is added under the conditions of being stirred at room temperature to 5, a large amount of solids, filtering drying is precipitated After obtain 1.8 grams of faint yellow solids.Faint yellow solid 300mg (0.8mmol, 1.0eq) is added in 25ml round-bottomed flask, according to Secondary addition 3mL DCM, 3 drip DMF, oxalyl chloride 300mg (2.4mmol, 3.0eq) are added dropwise under 0 DEG C of ice-water bath stirring condition, so After be warmed to room temperature reaction 2 hours, take out two drop reaction solutions be added methyl alcohol process be methyl esters, through LCMS detect reaction complete, decompression It is concentrated to get 320mg solid product JD1001-002-22, is saved backup under the conditions of nitrogen protection.

It is solid that 99mg (0.25mmol, 1.0eq) JD1001-002-22 is added under the conditions of nitrogen protection into 25mL round-bottomed flask Body, 3.2mL THF, 63mg (0.75mmol, 3.0eq) NaHCO₃, 140mg (0.75mmol, 3.0eq) 5- amino -3- fluoroform Base -2- cyanopyridine reacts 3 hours in 60 DEG C of heating stirrings, and (solvent: PE:EA=1:1) is completed in TLC detection reaction, is added Organic phase is obtained by extraction in 5ml water quenching reaction, EA, and concentration organic phase obtains solid, through HPLC (CH₃CN/H₂O=60:40 it) separates Purifying, obtains product J D1001-2139 about 16mg, yield 13% after freeze-drying.

¹H NMR:(DMSO,ppm,400MHz)δ1.47(6H,s),8.31(2H,s),8.38(1H,s),8.49(1H,d), 8.68 (1H, s), 9.12 (1H, d), 10.48 (1H, s) mass spectrums: [M+H]⁺549.

Embodiment 14 (preparation of compound JD1001-2156)

3.0g (18mmol, 1.0eq) 1- amino cyclobutane formate methyl esters salt is added under nitrogen protection into 100mL reactor 30mL (10.0V) THF is added into system, 3.0g (30.0mmol, 1.6eq) triethylamine is added dropwise at room temperature, then for hydrochlorate 0.3 gram of (2.3mmol, 0.13eq) DMAP is added, 5.05g (19.8mmol, 1.1eq) 4- bromobenzene sulfonyl chloride is then added dropwise, in room It is stirred to react under temperature 3 hours, TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops reaction, with full And NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses anhydrous slufuric acid Sodium dries, filters out organic phase, and organic phase is concentrated to give 6.3 grams of solids.

Solid is dissolved in 5ml THF, 15mL water is added, LiOH 1.2g (51.3mmol, 3.0eq) then is added, room temperature Under be stirred to react overnight, reaction monitored through LCMS, after the reaction was completed stop reaction, be concentrated under reduced pressure fall THF, be added into system dilute Hydrochloric acid (1N) adjusts pH value to 4, and a large amount of solids are precipitated, 2.1 grams of white solids are obtained by filtration, and dries.1g solid is taken, 3mL is dissolved in In (3V) DCM, 3 drop DMF are added, are cooled to 0 DEG C, 0.5ml oxalyl chloride is added dropwise at low temperature, is added dropwise and is warmed to room temperature Reaction 0.5 hour detects after taking a small amount of reaction solution that Derivatization of methanol is added through LCMS, and it is solid that end of reaction is concentrated under reduced pressure to give 1.5g Body product J D1001-002-23 is saved backup under nitrogen protection.

200mg (0.57mmol, 1.0eq) JD1001-002-23 is sequentially added under nitrogen protection into 25mL reactor, 3mL THF, 144mg (1.71mmol, 3eq) NaHCO₃, the fluoro- 5- amino pyrrole of 85.73mg (0.456mmol, 0.8eq) 2,3- bis- Pyridine is heated to 60 DEG C and is stirred to react 3 hours, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops Reaction, adds water quenching reaction, organic phase is obtained by extraction with ethyl acetate, then dry organic phase with anhydrous sodium sulfate, filtering is organic It is mutually concentrated to give solid 200mg, HPLC obtains 26mg white solid after purification.Yield is 15.4%

¹H NMR:(DMSO, ppm, 400MHz) δ 10.49 (s, 1H), 9.03 (s, 1H), 8.54-8.53 (s, 2H), 7.63- 7.61 (m, 4H), 2.50 (m, 2H), 2.46-2.43 (m, 2H), 1.78-1.75 (m, 1H), 1.66-1.63 (m, 1H) mass spectrum: 504[M+H]⁺

Comparative example 1 (preparation of compound JD1001-2103)

3.0g (20.0mmol, 1.0eq) 1- amino cyclobutane formate methyl esters is added under nitrogen protection into 100mL reactor 30mL (10.0V) THF is added into system, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, connects for hydrochloride Be added 0.3 gram of (2mmol, 0.1eq) DMAP, then be added dropwise 4.2g (22.2mmol, 1.1eq) 4- (Pentafluorosulfur) benzenesulfonyl chloride, is stirred at room temperature reaction 3 hours, and TLC detects raw material End of reaction (solvent: PE/EA=1/1, UV254) stops reaction, to be saturated NaHCO₃Quenching reaction, ethyl acetate extraction Organic phase is obtained, organic phase dries, filters out organic phase, organic phase concentration with 0.5N salt acid elution 2 times, then with anhydrous sodium sulfate Obtain 3.4 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, 4 grams of NaOH (5.0eq), heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase are free of product through detection, and water phase is added salt acid for adjusting pH value to 5, a large amount of solids are precipitated, are obtained by filtration 1.6 grams Faint yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of addition Derivatization of methanol through LCMS, end of reaction It is concentrated under reduced pressure to give 350mg solid product JD1001-002-16, is saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-16 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75.0mmol, 3.0eq) 3- amino -2- Trifluoromethyl pyridine is heated to 60 DEG C and is stirred to react 3 hours, TLC detection raw material end of reaction (solvent: PE/EA=1/1, UV254), stopping reaction, add water quenching reaction, organic phase is obtained by extraction with ethyl acetate, organic phase is dry with anhydrous sodium sulfate, Filtering, concentration organic phase obtains solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 38mg JD1001-2103, receive Rate 25%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.66 (1H, m), 1.81 (1H, m), 2.22 (2H, m), 7.94 (4H, m), 8.27 (1H, dd), 8.62 (1H, d), 8.69 (1H, d), 8.81 (1H, d), 9.97 (1H, s) mass spectrums: 526 [M+H]⁺

Comparative example 2 (preparation of compound JD1001-2124)

3.0g (20.0mmol, 1.0eq) 1- amino-cyclopropane methyl formate is added under nitrogen protection into 100mL reactor 30mL (10.0V) THF is added into system, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, connects for hydrochloride Be added 0.3 gram of (2mmol, 0.1eq) DMAP, then be added dropwise bis- (trifluoromethyl) the benzene sulphurs of 4.2g (22.2mmol, 1.1eq) 3,5- Reaction 3 hours is stirred at room temperature in acyl chlorides, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops Reaction, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses Anhydrous sodium sulfate dries, filters out organic phase, and organic phase is concentrated to give 3.4 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, NaOH4 grams (5.0eq), heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase through detection be free of product, water phase be added salt acid for adjusting pH value to 5, have solid precipitation, be obtained by filtration 1.6 grams it is light Yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of addition Derivatization of methanol through LCMS, end of reaction It is concentrated under reduced pressure to give 350mg solid product JD1001-002-12, is saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12,3.2mL is added under nitrogen protection into 25mL reactor (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, the fluoro- 5- aminopyridine of 0.38g (0.75mmol, 3.0eq) 2-, It is heated to 60 DEG C to be stirred to react 3 hours, TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, water quenching reaction is added, organic phase is obtained by extraction with ethyl acetate, then dry, filter out organic phase with anhydrous sodium sulfate, had Machine is mutually concentrated to give solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 41mg JD1001-2124.Yield 35%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.15 (2H, m), 1.38 (2H, m), 7.09 (1H, dd), 7.88 (1H, M), 8.14 (1H, d), 8.32 (2H, s), 8.38 (1H, s), 8.96 (1H, s), 9.74 (1H, s) mass spectrums: 472 [M+H]⁺

Comparative example 3 (preparation of compound JD1001-2125)

3.0g (20.0mmol, 1.0eq) 1- amino cyclobutane formate methyl esters is added under nitrogen protection into 25mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) the benzene sulphurs of 6.88g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in acyl chlorides, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops Reaction, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses Anhydrous sodium sulfate dries, filters out organic phase, is concentrated to give 2.8 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, 4 grams of (5.0eq) NaOH, heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase are free of product through detection, and water phase is added salt acid for adjusting pH value to 5, a large amount of solids are precipitated, are obtained by filtration 1.3 grams Faint yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of reaction solution that Derivatization of methanol is added through LCMS, instead It should finish and be concentrated under reduced pressure to give 350mg solid product JD1001-002-16, be saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12,3.2mL is added under nitrogen protection into 25mL reactor (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 2- amino-5-fluorine pyridine, It is heated to 60 DEG C to be stirred to react 3 hours, TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, adds water quenching reaction, organic phase is obtained by extraction with ethyl acetate, then dry, filter out organic phase with anhydrous sodium sulfate, be concentrated Solid is obtained, isolates and purifies to obtain 27mg JD1001-2125 through HPLC (CH3CN/H2O=60:40).Yield 18%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.23 (2H, m), 1.39 (2H, m), 7.65 (2H, m), 8.20 (1H, m), 8.26 (3H, s), 9.02 (1H, s), 9.78 (1H, s) mass spectrums: 472 [M+H]⁺

Comparative example 4 (preparation of compound JD1001-2126)

3.0g (20.0mmol, 1.0eq) 1- amino-cyclopropane methyl formate is added under nitrogen protection into 100mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) benzene sulfonyls of 4.2g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in chlorine, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses nothing Aqueous sodium persulfate dries, filters out organic phase, and organic phase is concentrated to give 3.4 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, 4 grams of NaOH (5.0eq), heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase through detection be free of product, water phase be added salt acid for adjusting pH value to 5, have solid precipitation, be obtained by filtration 3.5 grams it is light Yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of addition Derivatization of methanol through LCMS, end of reaction It is concentrated under reduced pressure to give 350mg solid product JD1001-002-12, is saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 2- amino -5- three Fluoromethylpyridin is heated to 60 DEG C and is stirred to react 3 hours, TLC detection raw material end of reaction (solvent: PE/EA=1/1, UV254), stop reaction, add water quenching reaction, organic phase, then dry, the mistake with anhydrous sodium sulfate are obtained by extraction with ethyl acetate Filter, organic phase are concentrated to give solid, isolate and purify to obtain 38mg JD1001-2126 through HPLC (CH3CN/H2O=60:40).Yield 26%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.27 (2H, m), 1.43 (2H, m), 7.78 (1H, d), 8.10 (1H, ), dd 8.20 (1H, s), 8.26 (2H, s), 8.59 (1H, dd), 9.03 (1H, s), 10.18 (1H, s) mass spectrums: 522 [M+H]⁺

Comparative example 5 (preparation of compound JD1001-2131)

3.0g (20.0mmol, 1.0eq) 1- amino-cyclopropane methyl formate is added under nitrogen protection into 100mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) benzene sulfonyls of 4.2g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in chlorine, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses nothing Aqueous sodium persulfate dries, filters out organic phase, and organic phase is concentrated to give 6.8 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, 4 grams of NaOH (5.0eq), heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase through detection be free of product, water phase be added salt acid for adjusting pH value to 5, have solid precipitation, be obtained by filtration 3.5 grams it is light Yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of addition Derivatization of methanol through LCMS, end of reaction It is concentrated under reduced pressure to give 350mg solid product JD1001-002-12, is saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12,3.2mL is added under nitrogen protection into 25mL reactor (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 2- amino -5- trifluoromethyl Pyridine is heated to 60 DEG C and is stirred to react 3 hours, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops It only reacts, adds water quenching reaction, organic phase is obtained by extraction with ethyl acetate, then dried, filtered with anhydrous sodium sulfate, organic phase is dense Contract to obtain solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 15mg JD1001-2131.Yield 11%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.17 (2H, d), 1.42 (2H, d), 7.79 (1H, d), 8.04 (1H, Dd), 8.32 (2H, s), 8.34 (1H, s) 8.66 (1H, d), 8.96 (1H, s), 10.04 (1H, s) mass spectrums: 522 [M+H]⁺

Comparative example 6 (preparation of compound JD1001-2111)

3.0g (20.0mmol, 1.0eq) 1- amino-cyclopropane methyl formate is added under nitrogen protection into 100mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) benzene sulfonyls of 4.2g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in chlorine, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses nothing Aqueous sodium persulfate dries, filters, and organic phase is concentrated to give 3.4 grams of solids.Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL is added Then (5V) water is added 4 grams of (5.0eq) NaOH, is heated to 60 DEG C and is stirred to react 2 hours, reaction is monitored through LCMS, and reaction is completed Stop reaction afterwards, be concentrated under reduced pressure ethyl alcohol, DCM extraction is added, organic phase is free of product through detection, and salt acid for adjusting pH is added in water phase It is worth 5, a large amount of solids is precipitated, 1.6 grams of faint yellow solids is obtained by filtration.

300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, 3eq is added dropwise at low temperature Oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of addition Derivatization of methanol through LCMS, end of reaction subtracts Pressure is concentrated to get 350mg solid product JD1001-002-12, saves backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 2- amino -5- three Fluoromethylpyridin is heated to 60 DEG C and is stirred to react 3 hours, TLC detection raw material end of reaction (solvent: PE/EA=1/1, UV254), stop reaction, add water quenching reaction, organic phase, then dry, the mistake with anhydrous sodium sulfate are obtained by extraction with ethyl acetate Filter, organic phase is concentrated to give solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 39mg JD1001-2111.Yield 31%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.23 (2H, dd), 1.44 (2H, dd), 8.09 (1H, dd), 8.29 (1H, s),8.32(2H,s),8.63(1H,d),8.78(1H,d),8.99(1H,s),9.98(1H,s).¹⁹F NMR:(DMSO, ppm, 400MHz) δ -61.47, -61.77. mass spectrum: 522 [M+H]⁺

Comparative example 7 (preparation of compound JD1001-2132)

3.0g (20.0mmol, 1.0eq) 1- amino-cyclopropane methyl formate is added under nitrogen protection into 100mL reactor 30mL (10.0V) THF is added into system, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, connects for hydrochloride Be added 0.3 gram of (2mmol, 0.1eq) DMAP, then be added dropwise bis- (trifluoromethyl) the benzene sulphurs of 4.2g (22.2mmol, 1.1eq) 3,5- Reaction 3 hours is stirred at room temperature in acyl chlorides, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops Reaction, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses Anhydrous sodium sulfate dries, filters, and organic phase is concentrated to give 3.4 grams of solids.

Solid is dissolved in ethyl alcohol (100mL, 15V), 30mL (5V) water is added, 4 grams of (5.0eq) NaOH, heating is then added It is stirred to react 2 hours to 60 DEG C, reaction is monitored through LCMS, stops reaction after the reaction was completed, is concentrated under reduced pressure ethyl alcohol, and DCM is added Extraction, organic phase are free of product through detection, and water phase is added salt acid for adjusting pH value to 5, a large amount of solids are precipitated, are obtained by filtration 1.6 grams Faint yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of reaction solution that Derivatization of methanol is added through LCMS, instead It should finish and be concentrated under reduced pressure to give 350mg solid product JD1001-002-12, be saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 2- amino -5- three Fluoromethylpyridin is heated to 60 DEG C and is stirred to react 3 hours, TLC detection raw material end of reaction (solvent: PE/EA=1/1, UV254), stop reaction, add water quenching reaction, organic phase, then dry, the mistake with anhydrous sodium sulfate are obtained by extraction with ethyl acetate Filter, organic phase is concentrated to give solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 24mg JD1001-2132, yield 19%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.17 (2H, d), 1.39 (2H, d), 7.27 (1H, dd), 7.93 (1H, D), 8.00 (1H, m), 8.32 (1H, s), 8.38 (1H, s), 9.21 (1H, s), 9.31 (1H, s) mass spectrums: 472 [M+H]⁺

Comparative example 8 (preparation of compound JD1001-2147)

3.0g (20.0mmol, 1.0eq) 1- amino cyclobutane formate methyl esters is added under nitrogen protection into 100mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) benzene sulfonyls of 4.2g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in chlorine, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses nothing Aqueous sodium persulfate dries, filters out organic phase, and organic phase is concentrated to give 3.4 grams of solids.Solid is dissolved in ethyl alcohol (100mL, 15V), is added Enter 30mL (5V) water, 4 grams of (5.0eq) NaOH are then added, is heated to 60 DEG C and is stirred to react 2 hours, reaction is monitored through LCMS, instead It should stop reaction after the completion, be concentrated under reduced pressure ethyl alcohol, DCM is added and extracts, organic phase is free of product through detection, and hydrochloric acid is added in water phase PH value is adjusted to 5, a large amount of solids are precipitated, 1.6 grams of faint yellow solids are obtained by filtration.

300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, 3eq is added dropwise at low temperature Oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of reaction solution that Derivatization of methanol is added through LCMS, reacts It finishes and is concentrated under reduced pressure to give 350mg solid product JD1001-002-12, saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, the chloro- 3- amino-of 0.38g (0.75mmol, 3.0eq) 2- 5- picoline is heated to 60 DEG C and is stirred to react 3 hours, TLC detection raw material end of reaction (solvent: PE/EA=1/1, UV254), stop reaction, add water quenching reaction, organic phase, then dry, the mistake with anhydrous sodium sulfate are obtained by extraction with ethyl acetate Filter, organic phase is concentrated to give solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 38mg JD1001-2147.Yield 20%.

¹H NMR:(DMSO, ppm, 400MHz) δ 1.72 (1H, m), 1.80 (1H, m), 2.20 (2H, m), 2.24 (3H, s), 2.46(2H,m),7.68(1H,d),8.02(1H,d),8.31(2H,s),8.44(1H,s),8.96(1H,s),9.11(1H,s)。 Mass spectrum: 516 [M+H]⁺

Comparative example 9 (preparation of compound JD1001-2155)

3.0g (20.0mmol, 1.0eq) 1- amino cyclobutane formate methyl esters is added under nitrogen protection into 100mL reactor Hydrochloride is charged with 30mL (10.0V) THF, 3.0g (30.0mmol, 1.5eq) triethylamine is added dropwise at room temperature, then 0.3 gram of (2mmol, 0.1eq) DMAP is added, bis- (trifluoromethyl) benzene sulfonyls of 4.2g (22.2mmol, 1.1eq) 3,5- are then added dropwise Reaction 3 hours is stirred at room temperature in chlorine, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), stops anti- It answers, to be saturated NaHCO₃Organic phase is obtained by extraction in quenching reaction, ethyl acetate, and organic phase is with 0.5N salt acid elution 2 times, then uses nothing Aqueous sodium persulfate dries, filters, and organic phase is concentrated to give 3.4 grams of solids.

Solid is dissolved in 100mL (15V) ethyl alcohol, 30mL (5V) water is added, 4 grams of (5.0eq) NaOH are then added, are heated to 60 DEG C are stirred to react 2 hours, and reaction is monitored through LCMS, stop reaction after the reaction was completed, are concentrated under reduced pressure ethyl alcohol, and DCM extraction is added Take, organic phase through detection be free of product, water phase be added salt acid for adjusting pH value to 5, a large amount of solids are precipitated, be obtained by filtration 1.6 grams it is light Yellow solid.300mg solid is taken, is dissolved in 3mL (3V) DCM, 3 drop DMF are added, is cooled to 0 DEG C, is added dropwise at low temperature 3eq oxalyl chloride is added dropwise and is warmed to room temperature reaction 2 hours, detects after taking a small amount of reaction solution that Derivatization of methanol is added through LCMS, instead It should finish and be concentrated under reduced pressure to give 350mg solid product JD1001-002-12, be saved backup under nitrogen protection.

0.1g (0.25mmol, 1.0eq) JD1001-002-12 is sequentially added under nitrogen protection into 25mL reactor, 3.2mL (10.0V) THF, 0.32g (0.75mmol, 3.1eq) NaHCO₃, 0.38g (0.75mmol, 3.0eq) 5,6- difluoro pyrrole Pyridine -3- amine is heated to 60 DEG C and is stirred to react 3 hours, and TLC detects raw material end of reaction (solvent: PE/EA=1/1, UV254), Stop reaction, adds water quenching reaction, organic phase is obtained by extraction with ethyl acetate, then dried, filtered with anhydrous sodium sulfate, organic phase It is concentrated to give solid, through HPLC (CH₃CN/H₂O=60:40 it) isolates and purifies to obtain 26mg JD1001-2155.Yield 17%.

¹H NMR:(DMSO, ppm, 400MHz) δ 9.95 (s, 1H), 8.93 (s, 1H), 8.30 (s, 1H), 8.19 (s, 2H), 7.96-7.95 (m, 2H), 2.50 (m, 2H), 2.26 (m, 2H), 1.86-1.83 (m, 1H), 1.70-1.62 (m, 1H)¹⁹F NMR: (DMSO, ppm, 400MHz) δ -61.70, -73.57, -94.30, -139.80. mass spectrum: 504 [M+H]⁺

Test example

In accordance with the following methods, activity test is carried out for the compound of above-described embodiment and comparative example:

(1) gained compound is dissolved in DMSO, is configured to the solution of 10mM, saved in -20 DEG C.

(2) logarithmic growth phase cell is collected, is counted, with complete medium again suspension cell, adjusts cell concentration to conjunction Suitable concentration (it is as follows that every hole inoculating cell number according to cell number optimizes preliminary result: Du-145 (human prostate cancer cell line, Androgen independence, AR-/PSA-, purchased from neat (Shanghai) the bioengineering Co., Ltd of match), 4000/hole；LNcap is (before people Column adenocarcinoma cell strain, androgen-dependent, AR+/PSA+ are purchased from the American Type Culture Collection committee, Chinese Academy of Sciences cell bank): 5000/hole), 96 orifice plates are inoculated with, every hole adds 100 μ l cell suspensions.Cell is at 37 DEG C, 100% relative humidity, 5%CO₂Culture It is incubated for 24 hours in case.

(3) untested compound is diluted to set respective action concentration with culture medium, cell is added by 25 holes μ l/. Compound effects final concentration is since 40 μM, 2 times of gradient dilutions, totally 6 concentration points (40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM), each 3 multiple holes of concentration point test.

(4) cell is placed in 37 DEG C, 100% relative humidity, 5%CO₂It is incubated for 72 hours in incubator.

(5) the every hole of 96 orifice plates is added to the CCK-8 detection reagent of 10 μ l, is mixed, in micropore after standing 1-2 hours at room temperature Light absorption signal (OD value) is measured on plate readout instrument FLUOstar Omega Multilabel Reader, and calculates inhibiting rate.

Specific Activity Results see the table below

By the comparison of above-mentioned activity data it is found that general formula compound of the present invention is the high anti-prostate cancer of a kind of activity Compound (referring to embodiment 1~14), compared with 1~9 compound represented of comparative example, the IC of DU-145 and LNcap₅₀Value is significant It reduces, in addition, anti-prostate cancer activity is also more significant compared with the Bicalutamide of current clinical use and the miscellaneous Shandong amine of grace.It is above-mentioned Statistics indicate that being had simultaneously by the compounds of this invention that trifluoromethyl and cyano replace for diseases such as prostate cancers on pyridine ring Excellent therapeutic effect.

Claims (7)

1. compound and its pharmaceutically acceptable salt that general formula (1) indicates,

Wherein, R₁With R₂It is identical or different, respectively indicate hydrogen atom or R₁With R₂It is formed together C_3-6Naphthenic base；

R₃Indicate hydrogen atom or C_1-8Alkyl；

R₄For substituted or unsubstituted C_6-10Aryl or heteroaryl, the heteroaryl are 5- or 6-membered monocycle base or are 5 yuan Or 6 unit monocycle and phenyl ring condensation made of bicyclic group.

2. compound as described in claim 1 and its pharmaceutically acceptable salt, wherein

R₄For the C at least one substituent group_6-10Aryl or heteroaryl,

The substituent group is selected from halogen atom, cyano, nitro, C_1-8Alkyl, C_1-8Alkoxy, halogenated C_1-8Alkyl, halogenated C_1-8Alkane Oxygroup, halogenated C_1-8Alkylthio group, halogenated C_1-8Alkyl sulphinyl, halogenated C_1-8Alkyl sulphonyl or pentafluoride-sulfanyl.

3. compound as described in claim 1 and its pharmaceutically acceptable salt, wherein R₄Are as follows:

In formula, n indicates 1~7 integer.

4. a kind of pharmaceutical composition, which is characterized in that containing compound of any of claims 1-3 or its pharmaceutically Acceptable salt and pharmaceutically acceptable carrier.

5. compound according to any one of claims 1 to 3 or its pharmaceutically acceptable salt are in preparation for preventing or controlling Treat the application in the drug of disease relevant to estrogen receptor activity.

6. application as claimed in claim 5, wherein the disease relevant to estrogen receptor activity includes: hormone-sensitive Property prostate cancer, hormone-refractory prostate cancer, benign prostatic hyperplasis, acne, hirsutism, whelk, dark sore and alopecia.

7. the preparation method for the compound that general formula (1) described in claim 1 indicates, which comprises the steps of:

Step (I) reacts compound (2) with oxalyl chloride or thionyl chloride in aprotic solvent, is formed compound (3)；

Step (II) reacts compound (3) with compound (4) in aprotic solvent, is formed compound (5)；

Step (III) in organic solvent reacts compound (5) with piperidines, is formed compound (6)；

Step (IV) in organic solvent reacts compound (6) with compound (7), is formed compound (1)；

R in the above formulas₁、R₂、R₃And R₄Definition with claim 1.