CN106810552B

CN106810552B - Thiohydantoin three-membered cyclic androgen receptor antagonist and use thereof

Info

Publication number: CN106810552B
Application number: CN201710057257.4A
Authority: CN
Inventors: 李志裕; 卞金磊; 徐熙; 葛饶玲
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2017-01-20
Filing date: 2017-01-20
Publication date: 2019-05-07
Anticipated expiration: 2037-01-20
Also published as: CN106810552A

Abstract

The present invention relates to field of medicinal chemistry, it using thio-hydantoin diindyl quinoline is skeleton with the compound (I) of anti-prostate cancer effect and (II) more particularly to one kind, including these compounds process for production thereof, these compounds can be with antagonism androgen receptor in molecular level, good anti-prostate cancer effect is shown in cellular level and animal level, can be used for preparing anti-tumor drug.

Description

Thio-hydantoin ternary and ring class androgen receptor antagonists and application thereof

Technical field

The present invention relates to field of medicinal chemistry, and in particular to a kind of having using thio-hydantoin diindyl quinoline as skeleton The compound of anti-prostate cancer effect, including these compounds process for production thereof, these compounds can be with antagonism heros in molecular level Hormone receptor shows good anti-prostate cancer effect in cellular level and animal level, can be used for preparing antineoplastic Object.

Background technique

Prostate cancer is the kinds of tumor for threatening American-European middle-aging male health, in recent years, with China's living standard It improves, life style changes correspondingly, and China's prostate cancer incidence is in rising trend.Huggins and Hodges is in nineteen forty-one head The secondary prostate cancer that reports is with androgen-dependent, as more and more evidences show that prostate cancer each stage is all swashed by hero Plain signal path is adjusted, and blocking this signal path is the available strategy for treating prostate cancer.One gamma therapy of clinic is mainly at present Castration of performing the operation and androgen deprivation carry out Androgen deprivation therapy (ADT).Although most of patients with prostate cancer are in early stage to ADT's Response rate is 80%-90%, and patient tolerance phenomenon often occurs after receiving to treat a period of time, leads to disease progression, develops and is Castration-resistant prostate cancer (CRPC).Therefore it is very urgent to develop effective castration-resistant prostate cancer therapeutic agent.

The key for finding and researching and developing the newtype drug for the treatment of CPRC be to probe into elaboration of tumour mechanism from molecular level and To the characterization of target.Studies have shown that the main reason for causing castration to resist is the androgen signal that androgen receptor (AR) is mediated The activation of axis additionally includes the regulation of other separate signal paths.Therefore block androgen synthesize and inhibit AR signal shaft at For the first choice strategy for the treatment of CRPC.

AR belongs to nuclear receptor superfamily, and molecular weight is about 110kDa, encodes 919 amino acid.AR is by several unique function Energy region composition, including the N- terminal region (NTD) with transcriptional activity, the bond area DNA (DBD) of Gao Baoshou, the end C- Ligand binding region (LBD) and hinge area (H) two organic functional areas AF-1 and AF-2 are located at the area NTD and LBD Domain can promote transcription to carry out.AR is a kind of ligand-dependent transcriptional factor, can be identified and be activated by native androgens, Key effect is all played to normal prostatic growth and the generation of prostate cancer and development.

Androgen receptor antagonists are divided into traditional AR antagonist and new A R antagonist, are divided into steroid and non-again by structure Steroid.The AR antagonist main function target clinically used at present is ligand binding region (LBD), can be competing with androgen Striving property combines the region, and induction H12 occurs wrong transposition and AR is inhibited to form excited conformation.

Steroid antagonist is similar with androgen structure, including megestrol acetate and cyproterone acetate etc..It has The characteristics of bioavilability is high, long half time, but due to hepatotoxicity wind agitation, cardiovascular side effects, such compound is in clinical application side Face is restricted.

Traditional non-steroidal AR antagonist Drogenil, Nilutamide, Bicalutamide are competitiveness AR antagonist, although its energy In conjunction with LBD cause H12 can not correct transposition, AR can still form dimer and enter nucleus and the faint combination of AREs, but by Decline in binding ability, the bioconjugation of AR and coactivator is suppressed, to promote AR, mortifier is combined together, final shadow Target gene transcription is rung, cell growth is inhibited.Traditional non-steroidal AR antagonist avoids the side effect of steroid antagonist, but also produces Raw some other problems.Such as the half-life short of Drogenil, patient needs 250mg tablet taken three times a day；The half of Nilutamide Although phase of declining is improved, but will lead to the side effects such as pneumonia；Bicalutamide tolerance is good and oral availability is high, but considers To safety issue, it is forbidden to use by many states.In addition, tradition AR antagonist can induce AR in use for some time Mutation, so that antagonist generates agonism to the AR after mutation.As Bicalutamide can be with 741 bit aminos of inducing receptor LBD Acid is sported leucine (W741L) by tryptophan, and activation AR promotes target gene transcription, and then patient is made to generate CRPC.Therefore it passes System AR antagonist can not effectively treat CRPC, and in response to this, new A R antagonist comes into being.

Summary of the invention

The invention discloses a kind of using thio-hydantoin ternary and ring is the novel androgen receptor antagonists of skeleton Close object.Pharmacodynamics test proves that the compound of the present invention has the competitive antagonist activity of androgen receptor, and there is treatment to dislike The effect of property tumour.

The compound of the present invention structure is as follows:

Wherein:

X represents carbon atom or nitrogen-atoms；

R₁Represent halogen or C₁~₄Halogenated alkyl；

R₂Represent hydrogen, halogen, cyano or hydroxyl；

R₂Also represent C₁~₄Alkyl, C₁~₄Alkoxy, C₁~₄Halogenated alkyl, cyclopropyl, imidazole radicals, pyrazolyl, triazole Base, tetrazole radical, morpholinyl, tetrahydrofuran base, piperidyl, piperazinyl, oxetanyl ,-C (O) R^a、-NR^aR^a、-COOR^a、- C(O)NR^aR^b、-C(S)NR^aR^b、-NHC(O)R^a、-NHSO₂R^aOr-SO₂NR^aR^b；Above-mentioned substituent group can be also optionally substituted by a hydroxyl group；Wherein R^a、R^bRespectively independently represent hydrogen, C₁~₃Alkyl, C₁~₃Hydroxyalkyl or R^a、R^bConnection forms nafoxidine base, piperidyl, morpholine Base, piperazinyl or N-methyl piperazinyl.

R₃Represent hydrogen or halogen；

R₄Represent hydrogen, halogen, cyano, hydroxyl or C₁~₄Alkoxy.

Wherein R₁It is preferred that representing halogen or trifluoromethyl；

R₂It is preferred that representing hydrogen, halogen, cyano, hydroxyl, cyclopropyl ,-NHC (O) R^a、-NHSO₂R^aOr-SO₂NR^aR^b；Wherein R^a、 R^bRespectively independently represent hydrogen, C₁~₂Alkyl, C₁~₂Hydroxyalkyl or R^a、R^bConnection forms nafoxidine base, piperidyl, morpholine Base, piperazinyl or N-methyl piperazinyl.

The compound of most preferably following any structure:

The preferred compound structure in part of the present invention is as follows:

The compound of the present invention can be prepared with following method:

The preparation of compound of Formula I:

The preparation of Compounds of formula II:

Wherein X, R₁、R₂、R₃、R₄It is as defined above.

Compound III is first and NBS reacts the bromine on benzyl position, and nucleophilic displacement of fluorine then occurs with acetamino diethyl malonate Reaction, then intermediate compound IV is obtained by hydrolyzing, preferred 60-110 DEG C of reaction temperature, reaction time preferred 10-12h, reaction dissolvent It is preferred that chloroform, ethyl alcohol, concentrated hydrochloric acid, water etc..

Compound IV reacts cyclization by Pictet-Spenler and obtains tetrahydroisoquinoline ring, is then esterified to obtain intermediate V, Preferred 30-80 DEG C of reaction temperature, reaction time preferred 8-18h, the preferred concentrated sulfuric acid of reaction dissolvent, acetic acid, methanol etc..

Compound V and compound VI cyclization, obtain preferred 25-60 DEG C of series compound I. reaction temperature, the reaction time is preferred 1-2h, preferred DMF of reaction dissolvent etc..Inorganic base or organic base should also be added in reaction, such as sodium hydroxide, potassium hydroxide, carbonic acid Potassium, sodium carbonate, triethylamine etc..

Compound VII obtains intermediate VIII by itrile group sodium borohydride reduction, preferred 30-80 DEG C of reaction temperature, reacts Time preferred 8-20h, the preferred methanol of reaction dissolvent, ethyl alcohol, tetrahydrofuran etc..

Compound VIII and compound VI cyclization, obtain preferred 25-60 DEG C of series compound II. reaction temperature, reaction time It is preferred that 1-2h, preferred DMF of reaction dissolvent etc..Inorganic base or organic base should also be added in reaction, as sodium hydroxide, potassium hydroxide, Potassium carbonate, sodium carbonate, triethylamine etc..

General formula I, II compound can be purified using common separation method, such as recrystallization, column chromatography.

The present invention also includes hydrate, stereoisomer, solvate and the pharmaceutically acceptable salt of these compounds Deng.They have pharmacological activity same as general formula I or II compound.

Compound of the present invention can add pharmaceutically acceptable carrier and common pharmaceutical formulation, such as piece is made Fragrance, sweetener, liquid or solid filler or diluent can be added in agent, capsule, pulvis, syrup, liquor, suspending agent, injection Etc. common medicinal supplementary materials.

The present invention has the advantages that it is raw materials used, reagent is simple and easy to get, reaction route is simple, high income；Structure novel； Its anti tumor activity in vitro is suitable better than positive control Enzalutamide or with it；The compounds of this invention is for DU145 cell Without obvious inhibiting effect (IC₅₀> 100 μM), thus alternatively the cytotoxicity of bright the compounds of this invention is less than Enzalutamide(IC₅₀It=46.81 μM, is shown in Table 1), is expected to be developed into anti-tumor drug.

Dosage used in the compound of the present invention clinic is 0.01mg-1000mg/ days, can also be according to the weight or dosage form of the state of an illness Difference deviate this range.

It is the pharmacology test and result of part of compounds of the present invention below:

(1) compound inhibits prostate gland cancer cell proliferation test:

Test objective: inhibited proliferation of the observation test compound to prostate gland cancer cell.DU145 is an AR feminine gender Cell, compound embody designed compound to the toxic effect of AR negative cells to its inhibited proliferation, can also Whether worked by AR from one aspect verifying compound.LNCaP cell is the AR positive cell of mutation, compound pair Its inhibited proliferation can embody designed compound to the inhibiting effect of the AR positive cell of mutation.

Test principle: with living cells metabolin reducing agent MTT, (full name is 3- (4,5- dimethylthiazole -2)-to MTT analytic approach 2,5- diphenyltetrazolium bromide bromides, trade name: thiazolyl blue) based on.MTT is yellow compound, is that a kind of receiving is hydrionic Dyestuff may act on the respiratory chain in living cells mitochondria, tetrazole ring under the action of succinate dehydrogenase and cromoci Cracking generates the first a ceremonial jade-ladle, used in libation crystallization of blue, the production quantity of first a ceremonial jade-ladle, used in libation crystallization (cell death then succinic acid only directly proportional to number of viable cells Dehydrogenase disappears, and cannot restore MTT).The first a ceremonial jade-ladle, used in libation crystallization that reduction generates can dissolve in DMSO, be measured using microplate reader Optical density OD value at 492nm, to reflect number of viable cells.

Test method: 1) inoculating cell: it is long to logarithmic growth phase to cell in the culture solution containing 10%FBS, use pancreatin It digests and is made into individual cells suspension, respectively with 7 × 1000, every hole DU145 or 1 × 10000LNCaP cell inoculation to 96 holes Plate；2) it is administered: 37 DEG C, 5%CO₂After culture for 24 hours, compound to be checked is dissolved and with culture solution respectively by dissolvedization with DMSO It closes object and is diluted to 0.1M/L, respectively with concentration gradient 10nM, 100nM, 1 μM, 10 μM, 100 μM of administrations, and set blank group and solvent Control group；3) 37 DEG C, 5%CO₂Continue to cultivate 72h；4) colour generation: every hole adds 20 μ l of MTT solution (5mg/ml), and it is small to continue incubation 4 When, careful inhale abandons culture supernatant in hole.150 μ l DMSO are added in every hole, vibrate 10 minutes, melt crystal sufficiently；5) compare Color: selection 492nm wavelength measures each hole absorbance value on enzyme linked immunological monitor, record as a result, calculate growth inhibition ratio, Draw growth inhibition curve.

The part of compounds of the present invention of table 1 is to prostate gland cancer cell (LNCaP, DU145) Proliferation Ability test result

Compound number	IC₅₀/μM(LNCaP)	IC₅₀/μM(DU145)
			CPU-101	5.32±0.60	143.1±0.89
CPU-102	26.97±0.93	>200
			CPU-103	32.60±2.22	67.07±2.66
CPU-104	9.91±3.20	>200
			CPU-105	10.44±0.61	>200
CPU-106	23.97±1.22	189.9±0.43
			CPU-107	21.71±2.19	78.47±1.10
CPU-108	33.14±0.86	>200
			CPU-109	27.77±5.63	>200
CPU-110	46.00±0.46	160.2±1.28
			CPU-111	38.40±1.67	>200
CPU-112	35.32±0.93	105.3±0.86
			CPU-113	30.87±0.57	113.7±1.10
CPU-201	77.25±3.78	58.07±3.10
			CPU-202	29.31±0.46	100.6±0.37
CPU-203	25.87±1.28	174.6±2.15
			CPU-204	48.47±0.49	>200
CPU-205	58.29±5.14	>200
			CPU-206	54.17±2.20	115.4±1.25
CPU-207	11.21±1.41	>200
			CPU-208	67.33±0.57	175.2±0.22
CPU-301	70.73±0.28	>200
			CPU-302	44.97±0.47	126.3±2.06
Enzalutamide	12.51±0.36	46.81±0.25

(2) antagonism test of the compound to androgen receptor:

Test objective: whether confirmation untested compound is to influence itself and target gene androgen response element by acting on AR It is transcribed in conjunction with influence.Can the intensity by detecting fluorescence inhibit androgen in conjunction with androgen receptor come compound of reaction, with Control group is compared, if experimental group fluorescence intensity reduces, illustrates that the compound of design can cause with androgen receptor competitive binding AR Dimerized cannot enter nucleus in conjunction with target gene response original part, to cannot cause subsequent transcription step.

Test principle: luciferase gene has become at present with one of widest reporter gene, luciferase (luciferase) luciferin can be catalyzed and is oxidized to oxyluciferin, this reaction is ATP dependence, heterocyclic optical Element is polyadenylation first, then by oxidative decarboxylation, generates AMP, CO₂, and pass through the transmitting of the fluorescein intermediate product of activation Then light measures the bioluminescence amount discharged in luciferin oxidation process by fluoremetry instrument, by data processing Meter calculates the content of luciferase in sample.

Test procedure: 1) (107 is thin for pMMTV-Luc, pcDNA3.1-AR and pRL-SV40 plasmid co-transfection COS-7 cell Born of the same parents/10cm plate), 37 DEG C, 5%CO₂Continue culture 24 hours；2) inoculating cell: with the tire ox of the processing containing 5% charcoal dextran Single cell suspension is made in serum free culture system liquid, respectively with every hole 5 × 10⁴A COS-7 cell inoculation is to 96 orifice plates；3) it is administered: with DMSO dissolves compound to be checked and by MDV3100, untested compound culture medium dissolved dilution to 0.1M/L, and with 10 μM of concentration Blank group and solvent control group is arranged in administration.Three multiple holes are arranged in each concentration；4) 37 DEG C, 5%CO₂Continue to cultivate 72h；5) Remove the culture medium in 96 orifice plates.Cell is cleaned with 1 × PBS, and discards PBS；6) 1 × PLB lytic cell: is added to culture Kong Zhong, every 20 μ L of hole, in the light and slow shaking culture plate 15min of room temperature；7) Activity determination: the LARII of balance to room temperature is added to In culture hole, every 100 μ L of hole, jog is mixed and is detected, and is postponed within 2 seconds when measurement, is read within 10 seconds；8) by balance to room temperatureReagent is added in culture hole, every 100 μ L of hole, and jog mixes detection and is same as above.It the results are shown in Table 2.

Antagonism test result of the part of compounds of the present invention of table 2 to androgen receptor

Compound number	Inhibiting rate % (10 μM)
		CPU-101	89.4
CPU-102	58.2
		CPU-103	29.0
CPU-104	91.9
		CPU-105	68.3
CPU-106	58.4
		CPU-107	21.8
CPU-108	23.4
		CPU-109	57.1
CPU-110	6.7
		CPU-111	13.1
CPU-112	2.1
		CPU-202	22.9
CPU-203	37.0
		CPU-204	39.3
CPU-207	70.51
		CPU-302	55.1
Enzalutamide	86.5

(3) compound is to androgen receptor competitive combination test:

Test objective: whether confirmation untested compound can specifically bind with androgen receptor, and the intensity combined. By detecting ability of the size of fluorescence polarization value come compound of reaction in conjunction with Selective Androgen Receptor.

Test principle: androgen receptor specific binding assays are under in vitro conditions, to detect androgen antagonist and AR The ability of aglucon binding domain selective binding.Under in vitro conditions, fluorescent ligand and AR aglucon binding domain are specifically bound, fluorescence Ligand freedom degree is low, and fluorescence polarization value is big；When compound and fluorescent ligand competitive binding AR aglucon binding domain, fluorescent ligand It is replaced, fluorescent ligand freedom degree increases, and fluorescence polarization value reduces.It is worth variation come calculating by detection fluorescence polarization Close the IC of object₅₀Value, according to IC₅₀Value judges the size of the affinity of AR antagonist and AR specific binding.It the results are shown in Table 3.

The part of compounds of the present invention of table 3 is to androgen receptor competitive combination test result

Compound number	IC₅₀/μM
		CPU-104	2.33±0.14
CPU-105	2.98±3.37
		CPU-106	4.49±3.03
Enzalutamide	1.35±1.13

(4) internal antitumor activity

Experimental method: growth animated period Human Prostate Cancer Cells are collected, cell suspension is aseptically prepared into, are inoculated with In nude mouse oxter.Nude Mice vernier caliper measurement transplantable tumor diameter, after tumour growth to will move after a certain size Object grouping, every group 5.Use the method for measurement knurl footpath, the antitumor effect of dynamic observation subject.Equal agent are given in blank control Measure hydroxypropyl-β-cyclodextrin；Compound group: tail vein injection continues 21 days once every other day.Tumor bearing nude mice is put to death after 21 days, and Separate tumor mass weighing.The data obtained carries out statistical procedures (t inspection), calculates tumour inhibiting rate.Meanwhile the tumor mass finally removed is clapped According to preservation picture.

The growth inhibition ratio of the LNCaP transplantable tumor of the part of compounds of the present invention of table 4

By table 4 as it can be seen that representative compound CPU-104 of the invention has inhibition well to mice-transplanted tumor LNCaP Effect has comparable inhibitory activity with Enzalutamide.

Specific embodiment

Embodiment 1

The synthesis (CPU102-CPU113) of compound of Formula I.

The preparation of 4- itrile group -3- fluorine bromobenzyl (2)

3- fluorine-4-nitrile toluene 1 (10g, 74.07mmol) is dissolved in chloroform 200mL, is added BPO 1g, stirring and dissolving, in batches It is added NBS (19.77g, 11.11mmol), flow back 12h, cooling, and reaction solution saturated sodium bicarbonate aqueous solution, which divides 3 times, to be washed, in right amount Washing 3 times, appropriate saturated common salt are washed 3 times, and evaporating solvent under reduced pressure, column chromatographs to obtain weak yellow liquid 14.00g, yield 88.79%.HRMS(ESI):m/z,calcd for C₈H₅BrFN 213.9678(M+H)⁺,found 213.9667。

The preparation of 2- (the fluoro- 4- cyanobenzyls of 3-) -2- acetamino diethyl malonate (3)

Ethyl alcohol 100mL is added in 250mL three-necked bottle, metallic sodium (0.76g, 33.16mmol) is added portionwise, stirring at normal temperature is extremely All dissolutions.It is added acetamino diethyl malonate (6.54g, 30.15mmol), stirring and dissolving, intermediate 2 is added (6.42g, 30.15mmol) is heated to flowing back, and reacts 18h, and reaction solution is cooled to room temperature, pours into 400mL ice water, stands analysis White solid out filters, dries to obtain white solid 7.50g, and yield 71.05%.HRMS (ESI): m/z, calcd for C₁₇H₁₉N₂O₅350.1321(M+H)⁺,found 350.1423。

The preparation of the fluoro- 4- of 2- (2- amino -2- carboxyethyl) benzoic acid (4)

Intermediate 3 (7.50g, 21.43mmol) is dissolved in concentrated hydrochloric acid 80mL, is heated to flowing back, and white opacity occurs, heat preservation 15h is stirred, reaction is stopped.It cools, filters, it is dry, obtain white solid 4.21g, yield 86.53%.HRMS(ESI):m/ z,calcd for C₁₀H₁₀NO₄ 228.0603(M+H)⁺,found 228.0606。

The preparation of 3- (the fluoro- 4- carbomethoxvphenvl of 3-) -2- aminopropanoate (5)

Intermediate 4 (10g, 44.05mmol) is dissolved in methanol 100mL, and ice salt bath is cooled to 0 DEG C hereinafter, being slowly added dropwise SOCl₂(10.48g, 88.10mmol) keeps the temperature 30min after dripping, is heated to flowing back, and reacts 8h, and TLC detects fully reacting. Solid, pumping rate, dry white solid 8.50g, yield 75.66% is precipitated in cooling.HRMS(ESI):m/z,calcd for C₁₂H₁₄NO₄ 256.0917(M+H)⁺,found 256.0914。

The preparation of N- ethoxycarbonyl -3- (the fluoro- 4- carbomethoxvphenvl of 3-) -2- aminopropanoate (6)

Intermediate 5 (4g, 15.69mmol) is dissolved in CH₂Cl₂In 40mL, it is passed through N₂Protection, ice-water bath are cooled to 0 DEG C, are added Pyridine (4.96g, 62.74mmol) maintains temperature at 0-5 DEG C, is slowly added dropwise ethyl chloroformate (1.70g, 15.69mmol), drips Reaction 4.5h is warmed to room temperature after adding, TLC detects fully reacting.100mL water, 50mL ethyl acetate is added in evaporating solvent under reduced pressure Divide 3 extractions, merges organic layer, 3 washings of 50mL saturated sodium-chloride point, organic layer anhydrous sodium sulfate drying.Filtering, filtrate Evaporated under reduced pressure obtains 3.08g white solid, yield 60.12%.HRMS(ESI):m/z,calcd for C₁₅H₁₈NO₆ 328.1119 (M+H)⁺,found 328.1123。

The preparation of N- ethoxycarbonyl -3,7- dimethoxy formoxyl -6- fluorine tetrahydroisoquinoline (7)

Intermediate 6 (3.83g, 11.71mmol) is added to CH₃COOH 12mL, dense H₂SO₄In the nitration mixture of 4mL, stir molten Solution.It is added paraformaldehyde (4.20g, 58.56mmol), 5h is stirred at room temperature, TLC detects fully reacting, stops reaction, adds 40mL water Dilution, 3 extractions of 30mL ethyl acetate point merge organic layer, and 30mL saturated sodium-chloride divides 3 washings, and anhydrous sodium sulfate is dry. Filtering, filtrate decompression are evaporated, and obtain white solid product 3.66g, yield 92.16%.HRMS(ESI):m/z,calcd for C₁₆H₁₈NO₆ 340.1118(M+H)⁺,found 340.1113。

The preparation of the fluoro- 3,7- dioctyl phthalate tetrahydroisoquinoline (8) of 6-

Intermediate 7 (5.00g, 14.74mmol) is dissolved in dense HCl 50mL, is heated to flowing back, and stirs 18h.It cools, Solid is precipitated, filters, dry white solid 2.65g, yield 75.23%.HRMS(ESI):m/z,calcd for C₁₁H₁₀NO₄240.0709(M+H)⁺,found 240.0706。

The preparation of 7- methoxycarbonyl base -6- fluorine tetrahydroisoquinoline -3- methyl formate (9)

Intermediate 8 (4.22g, 17.65mmol) is dissolved in methanol 50mL, and ice salt bath is cooled to 0 DEG C hereinafter, being slowly added dropwise SOCl₂(4.20g, 35.30mmol) keeps the temperature 30min after dripping, is heated to flowing back, and reacts 8h, and TLC detects fully reacting.Subtract Pressure is evaporated off partial solvent, is precipitated solid, pumping rate, dry white solid 3.71g, yield 78.73%.HRMS(ESI):m/z, calcd for C₁₃H₁₄NO₄ 268.0917(M+H)⁺,found 268.0918。

The preparation of compound CPU102

Intermediate 10 (0.23g, 1.00mmol) is dissolved in DMF 10mL, is added triethylamine (0.5g, 0.80mmol), is added Intermediate 9 (0.27g, 1.00mmol), is stirred at room temperature 2min, and TLC detects fully reacting, pours into 30mL ice water, 20mL acetic acid Points of 3 times extractions of ethyl ester, merge organic layer, are washed for 3 times with 40mL saturated common salt moisture, anhydrous sodium sulfate is dry.Filtering, removes under reduced pressure Solvent, column chromatograph to obtain 0.32g, and 69.11%.m.p.220-225 DEG C of yield；¹H-NMR (300MHz, DMSO-d₆):δ8.38(d,J =8.4Hz, 1H, Ar-H), 8.22 (s, 1H, Ar-H), 8.01 (d, J=8.4Hz, 1H, Ar-H), 7.96 (d, J=8.4Hz, 1H, ), Ar-H 7.42 (d, J=7.5Hz, 1H, Ar-H), 5.43 (s, 1H ,-CH-), 4.78 (m, 2H ,-CH₂-),3.88(s,3H,- CH₃), 3.40 (d, J=4.8Hz, 1H ,-CH₂-),3.34(s,1H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ 172.7,171.4,165.9,158.5,142.0,139.9,134.7,132.7,130.5,129.6,119.6,118.1, 115.8,115.7,115.6,114.4,105.0,72.5,53.8,51.5,28.1ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₃F₄N₃O₃S464.0616(M+H)⁺,found 464.0671；IR(KBr):3409,2957,1777,1628,1444, 1310,1268,1147,854,558cm^-1.

Embodiment 2

The preparation of compound CPU103

CPU102 (0.20g, 0.43mmol) is dissolved in tetrahydrofuran 5mL and 1N sodium hydrate aqueous solution 5mL, room temperature stirs 50min is mixed, TLC detection has been reacted, and decompression evaporates organic solvent, and 1N HCl is adjusted to pH=6, and solid is precipitated, and stands crystallization, is taken out Filter, column chromatograph to obtain 0.15g yellow solid, yield 76.53%.m.p.160-162℃；¹H-NMR (300MHz, DMSO-d₆):δ 13.26 (s, 1H ,-COOH), 8.13 (t, J=8.1Hz, 1H, Ar-H), 7.91 (m, 1H, Ar-H), 7.71 (d, J=8.1Hz, 1H, Ar-H), 7.56 (d, J=7.8Hz, 1H, Ar-H), 7.35 (d, J=8.1Hz, 1H, Ar-H), 5.46 (d, J=17.4Hz, 1H,-CH-),4.75-4.84(m,2H,-CH₂-),3.50-3.64(m,2H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆): δ172.7,171.1,165.1,158.9,142.9,139.9,134.7,132.7,130.9,129.6,119.6,118.1, 115.8,115.7,115.7,114.4,105.0,72.5,53.8,28.1ppm；HRMS(ESI):m/z,calcd for C₂₀H₁₁F₄N₃O₃S 450.0456(M+H)⁺,found 450.0530；IR(KBr):3413,2235,1718,1627,1443, 1313,1253,1140,804,558cm^-1.

Embodiment 3

The preparation of compound CPU104

CPU103 (0.10g, 0.22mmol) is dissolved in methylene chloride 10mL, ice bath, addition DMTMM (0.09g, 0.33mmol), 30min is stirred, is added triethylamine (0.09g, 0.88mmol), 30% methylamine alcohol solution (0.03g, 0.33mmol), 3h is stirred at room temperature, TLC detection has been reacted, and 1N HCl 20mL, 30mL ethyl acetate is added in evaporated under reduced pressure solvent Points of 3 times extractions merge organic layer, 3 washings of 30mL saturated sodium bicarbonate point, 30mL saturated common salt moisture 3 times washings, organic layer It is dry with anhydrous sodium sulfate, it filters, filtrate column chromatography obtains 0.08g yellow solid, yield 80.00%.m.p.329-333℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.38 (d, J=8.4Hz, 1H, Ar-H), 8.32 (s, 1H ,-NH-), 8.22 (s, 1H, Ar- ), H 8.02 (d, J=8.4Hz, 1H, Ar-H), 7.65 (d, J=8.4Hz, 1H, Ar-H), 7.33 (d, J=7.5Hz, 1H, Ar- ), H 5.39 (d, J=7.5Hz, 1H ,-CH-), 4.76 (m, J=7.5Hz, 2H ,-CH₂), 3.28 (t, J=7.5Hz, 2H ,- CH₂), 1.12 (t, J=6.9Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.8, 156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1,115.8,115.7, 114.6,105.0,72.5,53.4,28.1,26.7ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₄F₄N₄O₂S 463.0854(M+H)⁺,found463.0759；IR(KBr):3446,2963,1748,1649,1487,1310,1254,1140, 856,561cm^-1.

Embodiment 4

The preparation of compound CPU105

Methylamine alcohol solution is replaced with ethamine alcoholic solution, synthesizes compound in method identical with compound CPU104 CPU105, yield 76.41%.m.p.215-218℃；¹H-NMR(300MHz,DMSO-d₆):δ8.51(s,1H,Ar-H),8.41 (d, J=8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar- ), H 7.82 (d, J=7.5Hz, 1H, Ar-H), 5.39 (s, 1H ,-CH-), 5.32 (s, 2H ,-CH₂-),3.34(s,2H,-CH₂-), 3.29 (d, J=4.8Hz, 2H ,-CH₂), 1.15 (t, J=6.9Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ 172.7,171.4,167.5,156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6, 118.1,115.8,115.7,114.6,105.0,72.5,53.4,34.6,28.1,15.0ppm；HRMS(ESI):m/z,calcd for C₂₂H₁₆F₄N4O₂S 477.0930(M+H)⁺,found477.1002；IR(KBr):3403,2975,1766,1659, 1485,1312,1251,1141,849,558cm^-1.

Embodiment 5

The preparation of compound CPU106

Methylamine alcohol solution is replaced with n-propylamine, synthesizes compound CPU106 in method identical with compound CPU104, is received Rate 74.77%.m.p.279-281℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.41 (d, J=5.4Hz, 1H, Ar-H), 8.21 (s, 1H, Ar-H), 8.05 (d, J=7.8Hz, 1H, Ar-H), 7.69 (d, J=8.1Hz, 1H, Ar-H), 7.65 (d, J= 8.1Hz, 1H, Ar-H), 7.51 (d, J=7.8Hz, 1H, Ar-H), 7.09 (s, 1H ,-NH-), 5.26 (s, 2H ,-CH₂-),4.04 (q, J=6.0Hz, 1H ,-CH-), 3.21 (q, 2H ,-CH₂), 1.53 (q, J=6.0Hz, 2H ,-CH₂), 1.20 (q, J= 7.2Hz,2H,-CH₂), 0.91 (t, J=7.2Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7, 171.4,167.5,156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1, 115.8,115.7,114.6,105.0,72.5,53.4,41.5,28.1,23.1,11.2ppm；HRMS(ESI):m/z,calcd for C₂₃H₁₈F₄N₄O₂S 491.1087(M+H)⁺,found491.1160；IR(KBr):3415,2964,1743,1617,1440, 1313,1259,1159,820,558cm^-1.

Embodiment 6

The preparation of compound CPU107

Methylamine alcohol solution is replaced with isopropylamine, synthesizes compound CPU107 in method identical with compound CPU104, is received Rate 74.77%.m.p.240-243℃；¹H-NMR (300MHz, DMSO-d₆): δ 8.41 (d, J=5.4Hz, 1H, Ar-H), 8.21 (s, 1H ,-NH-), 8.05 (d, J=7.8Hz, 1H, Ar-H), 7.69 (d, J=8.1Hz, 1H, Ar-H), 7.51 (d, J= 7.8Hz,1H,Ar-H),7.09(s,1H,Ar-H),5.26(s,2H,-CH₂), 4.04 (q, J=6.0Hz, 1H ,-CH-), 3.21 (q,2H,-CH₂), 1.26 (d, J=6.0Hz, 3H ,-CH₃), 1.20 (d, J=7.2Hz, 3H ,-CH₃),0.87(m,1H,-CH-) ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.2,156.1,141.1,139.9,134.7,132.7, 129.8,128.1,122.1,119.6,118.1,115.8,115.7,114.6,105.0,72.5,53.4,45.2,28.1, 23.2,23.2ppm；HRMS(ESI):m/z,calcd for C₂₃H₁₈F₄N₄O₂S 489.1098(M-H)^-,found 489.1008；IR(KBr):3466,2971,1744,1656,1434,1314,1252,1126,897,558cm^-1.

Embodiment 7

The preparation of compound CPU108

Methylamine alcohol solution is replaced with cyclopropylamine, synthesizes compound CPU108 in method identical with compound CPU104, is received Rate 74.77%.m.p.279-282℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.51 (s, 1H, Ar-H), 8.41 (d, J= 8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 7.16 (s, 1H ,-CH-), 5.32 (s, 2H ,-CH₂-), 3.34(s,2H,-CH₂), 3.29 (d, J=4.8Hz, 2H ,-CH₂), 1.15 (q, J=6.9Hz, 1H ,-CH-), 1.24-1.48 (m,2H,-CH₂), 0.93 (t, J=7.2Hz, 2H ,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4, 167.2,156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1,115.8, 115.7,114.6,105.0,72.5,53.4,28.1,26.7,6.6,6.6ppm；HRMS(ESI):m/z,calcd for C₂₃H₁₆F₄N₄O₂S 489.1254(M+H)⁺,found489.1076；IR(KBr):3288,3070,1744,1660,1434, 1313,1253,1127,818,558cm^-1.

Embodiment 8

The preparation of compound CPU109

Methylamine alcohol solution is replaced with n-butylamine, synthesizes compound CPU109 in method identical with compound CPU104, is received Rate 74.77%.m.p.178-183℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.51 (s, 1H, Ar-H), 8.41 (d, J= 8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 7.16 (s, 1H ,-CH-), 5.32 (s, 2H ,-CH₂-), 3.34(s,2H,-CH₂), 3.29 (d, J=4.8Hz, 2H ,-CH₂), 1.15 (q, J=6.9Hz, 2H ,-CH₂-),1.24-1.48 (m,2H,-CH₂), 0.93 (t, J=7.2Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4, 167.5,156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1,115.8, 115.7,114.6,105.0,72.5,53.4,39.4,32.2,28.1,19.8,13.8ppm；HRMS(ESI):m/z,calcd for C₂₄H₂₀F₄N₄O₂S 505.1254(M+H)⁺,found 505.1320；IR(KBr):3415,2361,1711,1654, 1444,1311,1269,1148,1053,617cm^-1.

Embodiment 9

The preparation of compound CPU110

Methylamine alcohol solution is replaced with isobutyl amine, synthesizes compound CPU110 in method identical with compound CPU104, is received Rate 74.77%.m.p.136-140℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.41 (d, J=8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.12 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.64 (d, J=7.5Hz, 1H, Ar- H),7.25(m,1H,Ar-H),5.53(s,1H,-CH-),4.77(s,1H,-CH₂-),3.53(s,1H,-CH₂-),3.09(d,J =4.8Hz, 2H ,-CH₂), 2.25 (q, J=6.9Hz, 2H ,-CH₂), 1.01 (m, 1H ,-CH-), 0.90 (t, J=7.2Hz, 3H,-CH₃), 0.76 (t, J=7.2Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5, 156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1,115.8,115.7, 114.6,105.0,72.5,53.4,47.5,28.6,28.1,20.3,20.3ppm；HRMS(ESI):m/z,calcd for C₂₄H₂₀F₄N₄O₂S 503.1118(M-H)^-,found 503.1158；IR(KBr):3376,2961,1751,1659,1453, 1314,1272,1137,679,558cm-¹.

Embodiment 10

The preparation of compound CPU111

Methylamine alcohol solution is replaced with cyclohexylamine, synthesizes compound CPU111 in method identical with compound CPU104, is received Rate 74.77%.m.p.230-236℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.51 (s, 1H, Ar-H), 8.41 (d, J= 8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 7.16 (s, 1H ,-CH-), 5.32 (s, 2H ,-CH₂-), 3.34(s,2H,-CH₂), 3.29 (d, J=4.8Hz, 2H ,-CH₂), 1.15 (q, J=6.9Hz, 1H ,-CH-), 1.24-1.48 (m,2H,-CH₂), 0.93 (t, J=7.2Hz, 2H ,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4, 167.2,156.1,141.1,139.9,134.7,132.7,129.8,128.1,122.1,119.6,118.1,115.8, 115.7,114.6,105.0,72.5,53.4,51.6,32.3,32.3,28.1,25.7,24.8,24.8ppm；HRMS(ESI): m/z,calcd for C₂₆H₂₂F₄N₄O₂S 531.1432(M+H)⁺,found 531.1457；IR(KBr):3414,2930, 1750,1633,1426,1316,1258,1129,802,558cm^-1.

Embodiment 11

The preparation of compound CPU112

Methylamine alcohol solution is replaced with ethanol amine, synthesizes compound CPU112 in method identical with compound CPU104, is received Rate 74.77%.m.p.225-230℃；¹H-NMR (300MHz, DMSO-d₆): δ 8.51 (s, 1H, Ar-H), 8.41 (d, J= 8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 5.38 (s, 1H ,-CH-), 4.75 (s, 2H ,-CH₂-), 3.51(m,2H,-CH₂), 3.39 (d, J=4.8Hz, 2H ,-CH₂-),3.32(m,2H,-CH₂-),1.99(s,1H,-OH)ppm ；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5,156.1,141.1,139.9,134.7,132.7, 129.8,128.1,122.1,119.6,118.1,115.8,115.7,114.6,105.0,72.5,61.0,53.4,42.1, 28.1ppm；HRMS(ESI):m/z,calcd for C₂₂H₁₆F₄N₄O₃S 493.0954(M+H)⁺,found 493.0957；IR (KBr):3481,2386,1769,1670,1492,1310,1261,1149,646,559cm-¹.

Embodiment 12

The preparation of compound CPU113

Methylamine alcohol solution is replaced with isopropanolamine, synthesizes compound CPU113 in method identical with compound CPU104, Yield 74.77%.m.p.264-268℃；¹H-NMR (300MHz, DMSO-d₆): δ 8.39 (d, J=8.4Hz, 1H, Ar-H), 8.21 (d, J=8.4Hz, 2H, Ar-H), 8.02 (s, 1H ,-NH-), 7.69 (d, J=8.4Hz, 1H, Ar-H), 7.33 (s, 1H, Ar-H),5.39(s,1H,-CH-),4.78(s,2H,-CH₂-),4.68(s,1H,-CH₂), 3.80 (d, J=4.8Hz, 1H ,- CH₂-),3.38(s,1H,-CH-),3.29(s,1H,-OH),3.21(t,2H,-CH₂), 1.09 (d, 3H, J=8.4Hz ,-CH₃) ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5,156.1,141.1,139.9,134.7,132.7, 129.8,128.1,122.1,119.6,118.1,115.8,115.7,114.6,105.0,72.5,68.2,53.4,51.2, 28.1,21.1ppm；HRMS(ESI):m/z,calcd for C₂₃H₁₈F₄N₄O₃S 507.1134(M+H)⁺,found 507.1106；IR(KBr):3445,2964,1747,1661,1487,1312,1259,1138,802,559cm^-1.

Embodiment 13

The preparation of compound CPU301

The preparation of tetrahydroisoquinoline -3- carboxylic acid hydrochloride (12)

Phenylalanine (16.50g, 0.1mol) is dissolved in concentrated hydrochloric acid 150mL, 40% formalin of addition (70mL, 1mol), it is heated to reflux to solid and is completely dissolved, flow back 6h, filtering, and washing and drying obtains white solid 8.53g, yield 48.19%. m.p.247℃；¹H-NMR(300MHz,DMSO-d₆):δ10.07(s,1H,-COOH),7.27(s,4H,Ar-H),4.43-4.40 (m,2H,-CH₂-),3.81(s,1H,-CH-),3.45-3.10(m,2H,-CH₂-)；HRMS(ESI):m/z,calcd for C₁₀H₁₁NO₂178.1276(M+H)⁺,found 178.1230.

The preparation of compound (CPU301)

Intermediate 13 (0.18g, 1.00mmol) is dissolved in DMF 10mL, is added triethylamine (0.5g, 0.80mmol), in Mesosome 12 (0.18g, 1.00mmol), is stirred at room temperature 2min, and TLC detects fully reacting, pours into 30mL ice water, 20mL acetic acid second Points of 3 times extractions of ester, merge organic layer, are washed for 3 times with 40mL saturated common salt moisture, anhydrous sodium sulfate is dry.Filtering, removes under reduced pressure molten Agent, column chromatograph to obtain 0.19g, yield 56.38%.m.p.210-214℃；¹H-NMR(300MHz,DMSO-d₆):δ8.38(s,1H, ), Ar-H 8.15 (d, J=8.04Hz, 1H, Ar-H), 8.12 (d, J=3.6Hz, 1H, Ar-H), 7.74 (m, 1H, Ar-H), 7.65 (d, J=8.46Hz, 1H, Ar-H), 7.55 (m, 1H, Ar-H), 5.49 (t, J=18.66Hz, 1H ,-CH₂), 4.83 (dd, J= 5.43Hz,1H,-CH₂), 3.47 (t, J=2.97Hz, 1H ,-CH-) ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7, 171.4,160.8,141.2,134.0,130.4,128.6,127.5,126.9,126.3,125.7,121.3,110.8, 108.1,108.0,72.5,53.8,28.1ppm；HRMS(ESI):m/z,calcd for C₁₈H₁₂FN₃OS 338.0097(M+H )⁺,found338.0085；IR(KBr):3125,2231,1752,1617,1525,1400,1347,1069,860,515cm^-1.

Embodiment 14

The preparation of compound CPU302

The preparation of 7- nitro tetrahydroisoquinoline -3- carboxylic acid hydrochloride (14)

It is poured into concentrated sulfuric acid 12.5mL in 100mL three-necked bottle, is cooled to -10 DEG C, tetrahydroisoquinoline -3- carboxylic acid 12 is added portionwise (2.5g, 14.11mmol) maintains the temperature at -10 DEG C hereinafter, potassium nitrate (1.48g, 14.74mmol) is dissolved in the concentrated sulfuric acid simultaneously It is stored refrigerated in 5mL.To 12 concentrated sulfuric acid solution after mixing evenly, slowly instill potassium nitrate concentrated sulfuric acid solution, duration requirement In 1.5h or more, reaction temperature is kept for -10 DEG C or less.After being added dropwise, be warmed to room temperature stirring 1h, then by reaction solution incline to In 100mL ice water, ammonium hydroxide adjusts pH to neutrality, filters, washes filter cake, obtain yellow solid 1.41g, yield 45.13%；¹H- NMR(300MHz,D₂O): δ 8.35 (d, 1H, Ar-H), 8.17 (dd, 1H, Ar-H), 7.63 (d, 1H, Ar-H), 4.61-4.32 (m,3H),3.62–3.21(m,2H)ppm；HRMS(ESI):m/z,calcd for C₁₀H₁₀N₂O₄ 223.0698(M+H)⁺, found 223.0667.

The preparation of 7- nitro tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (15)

Intermediate 14 (1.00g, 4.48mmol) is dissolved in methanol 30mL, ice salt bath is cooled to -10 DEG C hereinafter, chlorine is added dropwise Change sulfoxide (1.07g, 8.96mmol), equality of temperature stirs 30min, and then heating makes reaction solution back flow reaction 14h, cools down, and decompression is steamed Except partial solvent, solid is precipitated, filters, is dried to obtain yellow solid 0.52g, yield 48.72%.(product directly carries out lower step Reaction) HRMS (ESI): m/z, calcd for C₁₁H₁₂N₂O₄ 237.0819(M+H)⁺,found 237.0821。

The preparation of compound CPU302

Intermediate 13 (0.18g, 1.00mmol) is dissolved in DMF 10mL, is added triethylamine (0.5g, 0.80mmol), intermediate Body 15 (0.24g, 1.00mmol), is stirred at room temperature 2min, and TLC detects fully reacting, pours into 30mL ice water, 20mL ethyl acetate Points of 3 times extractions, merge organic layer, are washed for 3 times with 40mL saturated common salt moisture, anhydrous sodium sulfate is dry.Filtering, removes under reduced pressure molten Agent, column chromatograph to obtain 0.27g, yield 70.68%.m.p.212-214℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.15 (d, J= 8.04Hz, 1H, Ar-H), 8.12 (d, J=3.6Hz, 1H, Ar-H), 7.74 (m, 1H, Ar-H), 7.65 (d, J=8.46Hz, 1H, ), Ar-H 7.55 (m, 1H, Ar-H), 5.38 (d, J=18.66Hz, 1H ,-CH-), 4.75 (m, 2H ,-CH₂), 3.30 (t, J= 2.97Hz,2H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,160.8,150.4,141.2, 136.5,135.0,134.0,129.8,122.1,121.7,121.3,110.8,108.1,108.0,72.5,52.8, 28.1ppm；HRMS(ESI):m/z,calcd for C₁₈H₁₁FN₄O₃S 383.0678(M+H)⁺,found 383.0545；IR (KBr):3123,1750,1615,1434,1400,1310,1280,1158,765cm^-1.

Embodiment 15

The preparation of compound CPU303

Intermediate 13 (0.18g, 1.00mmol) is dissolved in DMF 10mL, is added triethylamine (0.5g, 0.80mmol), intermediate 9 2min is stirred at room temperature in (0.28g, 1.00mmol), and TLC detects fully reacting, pours into 30mL ice water, 20mL ethyl acetate point 3 Secondary extraction, merges organic layer, is washed for 3 times with 40mL saturated common salt moisture, and anhydrous sodium sulfate is dry.Filtering, evaporating solvent under reduced pressure, column Chromatograph to obtain 0.29g, yield 70.22%.m.p.228-230℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.14 (t, J=5.4Hz, 1H, Ar-H), 7.95 (d, J=5.4Hz, 1H, Ar-H), 7.71 (d, J=7.8Hz, 1H, Ar-H), 7.56 (d, J=8.1Hz, 1H, Ar-H), 7.41 (d, J=8.1Hz, 1H, Ar-H), 5.41 (m, 1H ,-CH-), 4.78 (m, 2H ,-CH₂-),3.87(s, 3H,-CH₃), 3.37 (q, J=6.0Hz, 2H ,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4, 165.9,160.8,158.5,142.0,141.2,134.0,30.5,129.6,121.3,115.6,114.4,110.8,108.1, 108.0,72.5,53.8,51.5,28.1ppm；HRMS(ESI):m/z,calcd for C₂₀H₁₃F₂N₃O₃S 414.0747(M+ H)⁺,found414.0713；IR(KBr):3482,2236,1662,1444,1367,1200,1140,890,558cm^-1.

Embodiment 16

The preparation of compound CPU304

CPU303 (0.20g, 0.48mmol) is dissolved in tetrahydrofuran 5mL and 1N sodium hydrate aqueous solution 5mL, room temperature is stirred 50min is mixed, TLC detection has been reacted, and decompression evaporates organic solvent, and 1N HCl is adjusted to pH=6, and solid is precipitated, and stands crystallization, is taken out Filter, column chromatograph to obtain 0.15g yellow solid, yield 77.72%.m.p.246-248℃；¹H-NMR(300MHz,DMSO-d₆):δ 13.26 (s, 1H ,-COOH), 8.13 (t, J=8.1Hz, 1H, Ar-H), 7.91 (m, 1H, Ar-H), 7.71 (d, J=8.1Hz, 1H, Ar-H), 7.56 (d, J=7.8Hz, 1H, Ar-H), 7.35 (d, J=8.1Hz, 1H, Ar-H), 5.46 (d, J=17.4Hz, 1H,-CH-),4.84-4.75(m,2H,-CH₂-),3.64-3.50(m,2H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆): δ172.7,171.4,165.1,160.8,158.9,142.9,141.2,134.0,130.9,129.6,121.3,115.7, 114.4,110.8,108.1,108.0,72.5,53.8,28.1ppm；HRMS(ESI):m/z,calcd for C₁₉H₁₁F₂N₃O₃S400.5090(M+H)⁺,found 400.0489；IR(KBr):3531,2234,1760,1628,1449, 1376,1251,1139,896,558cm^-1.

Embodiment 17

The preparation of compound CPU305

CPU304 (0.10g, 0.25mmol) is dissolved in methylene chloride 15mL, ice bath, addition DMTMM (0.04g, 0.37mmol), 30min is stirred, is added triethylamine (0.13g, 1.25mmol), methylamine alcohol solution (0.05g, 0.49mmol), room Temperature stirring 3h, TLC detection has been reacted, and evaporated under reduced pressure solvent is added 3 extractions of 1N HCl 20mL, 30mL ethyl acetate point, closes And organic layer, 3 washings of 30mL saturated sodium bicarbonate point, 30mL saturated common salt moisture 3 times washings, organic layer anhydrous sodium sulfate It is dry, it filters, filtrate column chromatography obtains 0.08g yellow solid, yield 78.77%.m.p.306-309℃；¹H-NMR(300MHz, DMSO-d₆): δ 8.25 (s, 1H ,-NH-), 8.13 (t, J=8.4Hz, 1H, Ar-H), 7.69 (m, 2H, Ar-H), 7.55 (d, J= 8.4Hz, 1H, Ar-H), 7.32 (d, J=8.4Hz, 1H, Ar-H), 5.33 (d, 1H ,-CH-), 4.75 (m, 2H ,-CH₂-),3.33 (s,2H,-CH₂), 2.77 (d, J=8.4Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4, 167.8,160.8,156.1,141.2,141.1,134.0,129.8,128.1,122.1,121.3,114.6,110.8, 108.1,108.0,72.5,53.4,28.1,26.7ppm；HRMS(ESI):m/z,calcd for C₂₀H₁₄F₂N₄O₂S413.0811(M+H)⁺,found 413.0884；IR(KBr):3424,2244,1748,1620,1450, 1380,1247,1168,819,512cm^-1.

Embodiment 18

The preparation of compound CPU306

Methylamine alcohol solution is replaced with ethamine alcoholic solution, synthesizes compound in method identical with compound CPU305 CPU306, yield 79.73%.m.p.274-276℃；¹H-NMR (300MHz, DMSO-d₆):δ8.51(s,1H,Ar-H),8.41 (d, J=8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar- ), H 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 5.38 (d, 1H ,-CH-), 4.75 (s, 2H ,- CH₂-),3.99(m,1H,-CH₂-),3.27(m,2H,-CH₂-),1.24(m,1H,-CH₂), 1.13 (d, 3H, J=8.4Hz ,- CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5,160.8,156.1,141.2,141.1, 134.0,129.8,128.1,122.1,121.3,114.6,110.8,108.1,108.0,72.5,53.4,28.1,34.6, 15.0ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₆F₂N₄O₂ 427.0966(M+H)⁺,found 427.1038；IR (KBr):3412,2244,1751,1617,1448,1377,1308,1253,826,612cm^-1.

Embodiment 19

The preparation of compound CPU307

Methylamine alcohol solution is replaced with ethanol amine, synthesizes compound CPU307 in method identical with compound CPU305, is received Rate 74.97%.m.p.208-210℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.23 (s, 1H ,-NH-), 8.12 (t, J= 8.4Hz, 1H, Ar-H), 7.73 (m, 2H, Ar-H), 7.55 (t, J=8.4Hz, 1H, Ar-H), 7.32 (s, 1H, Ar-H), 5.37 (d, J=4.8Hz, 1H ,-CH-), 4.77 (t, J=4.8Hz, 2H ,-CH₂), 4.66 (s, 1H ,-OH), 3.50 (t, J= 4.8Hz,2H,-CH₂-),3.36(m,4H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5, 160.8,156.1,141.2,141.1,134.0,129.8,128.1,122.1,121.3,114.6,110.8,108.1, 108.0,72.5,61.0,53.4,42.1,28.1ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₆F₂N₄O₃S 443.0911(M+H)⁺,found 443.0984；IR(KBr):3408,2243,1751,1617,1448,1377,1254, 1164,825,611cm^-1.

Embodiment 20

The preparation of compound CPU308

Methylamine alcohol solution is replaced with isopropanolamine, synthesizes compound CPU308 in method identical with compound CPU305, Yield 75.89%.m.p.230-234℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.51 (s, 1H, Ar-H), 8.41 (d, J= 8.4Hz, 1H, Ar-H), 8.33 (s, 1H ,-NH-), 8.06 (d, J=8.4Hz, 1H, Ar-H), 7.95 (s, 1H, Ar-H), 7.82 (d, J=7.5Hz, 1H, Ar-H), 7.64-7.72 (m, 1H, Ar-H), 5.38 (s, 1H ,-CH-), 4.75 (s, 2H ,-CH₂-), 4.66(s,1H,-OH),3.78(m,2H,-CH₂), 3.36 (d, J=4.8Hz, 1H ,-CH-), 3.31 (m, 2H ,-CH₂-),1.08 (d, 3H, J=8.4Hz ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ172.7,171.4,167.5,160.8,156.1, 141.2,141.1,134.0,129.8,128.1,122.1,121.3,114.6,110.8,108.1,108.0,72.5,68.2, 53.4,51.2,28.1,21.1ppm；HRMS(ESI):m/z,calcd for C₂₂H₁₈F₂N₄O₃S 457.1067(M+H)⁺, found 457.1140；IR(KBr):3448,2236,1769,1615,1445,1311,1249,1164,1089,805, 512cm^-1.

Embodiment 21

The synthesis (CPU401-CPU409) of general formula Ia compound

The preparation of 7- amino -1,2,3,4- tetrahydroisoquinoline -3- methyl formate hydrochloride (17)

7- nitro -1,2,3,4- tetrahydroisoquinoline -3- methyl formate hydrochloride (0.10g, 0.37mmol) is dissolved in 10mL In methanol, Pd/C is added, reacts at room temperature 10min, filters and removes palladium carbon, filtrate rotation obtains sepia solid 0.07g except solvent, produces Rate 80.0%.m.p.236℃；HRMS(ESI):m/z,calcd for C₁₁H₁₄N₂O₂ 207.1128(M+H)⁺,found 207.1129。¹H-NMR(300MHz,DMSO-d₆):δ3.52-3.01(m,2H),3.80(s,3H,-OCH₃),4.51-4.22(m, 3H),7.43-7.10(m,3H)。

The preparation of N- tertbutyloxycarbonyl -7- nitro tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (18)

7- nitro tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (7.01g, 0.03mol) is dissolved in 100mL methylene chloride, (3.00g, 0.03mol) triethylamine is added to 0 DEG C in ice bath, is added dropwise (13.08g, 0.06mmol) (Boc)₂O, Mao great Liang bubble, TLC detection has been reacted after reacting 3h under room temperature, and evaporated under reduced pressure, column chromatographs to obtain 5.60g yellow oil, yield 55.6%. HRMS(ESI):m/z,calcd for C₁₆H₂₀N₂O₆ 337.1394(M+H)⁺,found 337.1396。

The preparation of N- tertbutyloxycarbonyl -7- amino tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (19)

N- tertbutyloxycarbonyl 7- nitro tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (5.60g, 16.67mmol) is dissolved in In 56mL methanol, 0.56g Pd/C is added, normal temperature and pressure is passed through hydrogen, reacts 6h, and TLC detects fully reacting, filters, and filtrate subtracts Pressure is evaporated, and obtains 4.51g off-white powder, yield 88.4%.HRMS(ESI):m/z,calcd for C₁₆H₂₂N₂O₄ 307.1652 (M+H)⁺,found 307.1653。

The preparation of N- tertbutyloxycarbonyl -7- acetamido tetrahydroisoquinoline -3- carboxylate methyl ester (20a)

N- tertbutyloxycarbonyl 7- amino tetrahydroisoquinoline -3- carboxylate methyl ester hydrochloride (0.61g, 1.99mmol) is dissolved in 10mL In methylene chloride, ice bath is added dropwise (0.80g, 7.96mmol) triethylamine, (0.31g, 3.96mmol) chloroacetic chloride is dissolved in 2mL bis- It in chloromethanes, is slowly dropped in reaction solution, reacts at room temperature 3h, TLC detects fully reacting, and 30mL water, 20mL dichloromethane is added Alkane extract three times, merge organic layer, 30mL saturated sodium bicarbonate wash three times, 30mL saturated common salt water washing three times, organic layer It is dry with anhydrous sodium sulfate, it filters, filtrate decompression is evaporated, and obtains 0.62g yellow solid, yield 89.9%.HRMS(ESI):m/z, calcd for C₁₈H₂₄N₂O₅ 349.1758(M+H)⁺,found 349.1760。

The preparation of 7- acetamido tetrahydroisoquinoline -3- carboxylate methyl ester (21a)

N- tertbutyloxycarbonyl 7- acetamido tetrahydroisoquinoline -3- carboxylate methyl ester (0.62g, 1.78mmol) is dissolved in 12mL bis- In chloromethanes, trifluoracetic acid 1.24mL is added dropwise under ice bath, is stirred overnight, evaporating solvent under reduced pressure, adds 10mL water, saturated sodium bicarbonate It is adjusted to pH=7,30mL ethyl acetate extracts three times, merges organic layer, and 50mL saturated salt solution washes twice, and anhydrous sodium sulfate is dry It is dry, it filters, filtrate decompression is evaporated to obtain brown solid 0.29g, yield 65.9%.HRMS(ESI):m/z,calcd forC₁₃H₁₆N₂O₃ 249.1234(M+H)⁺,found 249.1236。

The preparation of compound CPU401

Intermediate 15 (0.27g, 1.00mmol) is dissolved in 10mL anhydrous DMF, is added triethylamine (0.10g, 1.00mmol) Clarify solution, at 28 DEG C into reaction solution be added 5- isothiocyanates -3- trifluoromethyl -2- itrile group pyridine 16 (0.23g, 1.00mmol), temperature slightly rises.2min is stirred at room temperature, TLC detects fully reacting, pours into 20mL ice water, 20mL methylene chloride Extraction 3 times merges organic layer, is washed 3 times with 40mL saturated common salt, and anhydrous sodium sulfate is dry.Filtering, evaporating solvent under reduced pressure, column Chromatograph to obtain 0.15g bright yellow solid, yield 38.6%.m.p.134-136℃；¹H-NMR(300MHz,DMSO-d₆):δ9.18(d, J=1.6Hz, 1H, Ar-H), 8.72 (d, J=1.6Hz, 1H, Ar-H), 8.38 (d, J=1.5Hz, 1H, Ar-H), 8.16-8.13 (m, 1H, Ar-H), 7.66 (d, J=8.6Hz, 1H, Ar-H), 5.55 (d, J=18.2Hz, 1H ,-CH₂-),4.94-4.90(m, 1H ,-CH-), 4.87 (d, J=18.2Hz, 1H ,-CH₂-),3.56-3.43(m,2H,-CH₂-)ppm；¹³C NMR(75MHz, DMSO-d₆):δ177.5,172.0,153.5,147.6,135.5,135.4,133.2,130.7,129.6,123.3,119.7, 117.3,114.3,113.6,110.7,58.3,45.6,28.5ppm；HRMS(ESI):m/z,calcd for C₁₈H₁₀F₃N₅O₃S 433.0456(M-H)^-,found 433.0455；IR(KBr):3448,1762,1499,1491,1348,1309,1257, 1172,1146,1058,743,641cm^-1.

Embodiment 22

The preparation of compound CPU402

Intermediate 15 is replaced with intermediate 17, synthesizes compound CPU402 in method identical with compound CPU401, is received Rate 27.9%.m.p.206-208℃；¹H-NMR(300MHz,DMSO-d₆):δ9.16(s,1H,Ar-H),8.72(s,1H,Ar- ), H 6.97-7.02 (m, 1H, Ar-H), 6.48 (s, 2H, Ar-H), 5.22 (d, J=17.1Hz, 1H ,-CH₂-),5.11(s, 2H,-NH₂),4.80-4.60(m,2H,-CH₂), 4.54 (d, J=17.1Hz, 1H ,-CH₂-),3.11(s,2H,-CH₂-)ppm ；¹³C-NMR(75MHz,DMSO-d₆):δ177.5,172.0,153.5,147.6,135.5,135.4,133.2,130.7, 129.6,123.3,119.7,117.3,114.3,113.6,110.7,58.3,45.7,28.5ppm；HRMS(ESI):m/z, calcd for C₁₈H₁₂F₃N₅OS 403.0715(M-H)^-,found 403.0710；IR(KBr):3448,1763,1624, 1508,1483,1376,1271,1175,1140,1058cm^-1.

Embodiment 23

The preparation of compound CPU403

Intermediate 15 is replaced with intermediate 21a, synthesizes compound CPU403 in method identical with compound CPU401, is received Rate 22.9%.m.p.230-232℃；¹H-NMR(300MHz,DMSO-d₆):δ10.01(s,1H,-NH-),9.18(s,1H,Ar- ), H 8.73 (s, 1H, Ar-H), 7.60 (s, 1H, Ar-H), 7.42 (s, 1H, Ar-H), 7.27 (s, 1H, Ar-H), 5.37 (d, J= 15.7Hz,1H,-CH₂), 4.84-4.82 (m, 1H ,-CH-), 4.70 (d, J=15.7Hz, 1H ,-CH₂-),3.26-3.24(m, 2H,-CH₂-),2.04(s,3H,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ177.7,171.9,168.3,153.5, 138.2,135.5,135.4,133.2,130.8,129.4,128.8,125.2,123.3,118.1,116.5,114.2,57.8, 45.7,28.7,23.9ppm；HRMS(ESI):m/z,calcd for C₂₀H₁₄F₃N₅O₂S 445.0820(M-H)^-,found 445.0817；IR(KBr):3422,3373,2926,1769,1459,1289,1172,1142cm^-1.

Embodiment 24

The preparation of compound CPU404

Chloroacetic chloride, first synthetic intermediate 20b and 21b, then with side identical with compound CPU401 are replaced with propionyl chloride Method synthesizes compound CPU404, yield 26.7%.m.p.152-154℃；¹H-NMR(300MHz,DMSO-d₆):δ9.94(s, 1H ,-NH-), 9.18 (d, J=1.5Hz, 1H, Ar-H), 8.73 (d, J=1.5Hz, 1H, Ar-H), 7.63 (s, 1H, Ar-H), 7.45 (d, J=8.0Hz, 1H, Ar-H), 7.27 (d, J=8.0Hz, 1H, Ar-H), 5.35 (d, J=17.4Hz, 1H ,-CH₂-), 4.87-4.81 (m, 1H ,-CH-), 4.71 (d, J=17.4Hz, 1H ,-CH₂-),3.27-3.24(m,2H,-CH₂-),2.33(q, 2H, J=7.4Hz ,-CH₂), 1.09 (t, J=7.4Hz, 3H ,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ177.7, 172.0,171.9,153.5,138.2,135.5,135.4,133.2,130.7,129.4,128.9,125.1,123.3, 118.2,116.6,114.2,57.8,45.7,29.4,28.7,9.6ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₆F₃N₅O₂S 459.0977(M-H)^-,found459.0972；IR(KBr):3432,1745,1507,1498,1491, 1459,1313,1285,1173,1145,1059cm^-1.

Embodiment 25

The preparation of compound CPU405

Chloroacetic chloride, first synthetic intermediate 20c and 20c, then with side identical with compound CPU401 are replaced with butyl chloride Method synthesizes compound CPU405, yield 45.8%.m.p.120-128℃；¹H-NMR(300MHz,DMSO-d₆):δ9.94(s, 1H,-NH-),9.19(s,1H,Ar-H),8.73(s,1H,Ar-H),7.64(s,1H,Ar-H),7.44(m,1H,Ar-H),7.28 (s, 1H, Ar-H), 5.34 (d, J=17.2Hz, 1H ,-CH₂), 4.83-4.70 (m, 1H ,-CH-), 4.70 (d, J=17.2Hz, 1H,-CH₂-),3.26-3.24(m,2H,-CH₂-),2.28(s,2H,-CH₂-),1.62-1.60(m,2H,-CH₂-),0.92(s, 3H,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ177.7,171.9,171.1,153.5,139.4,138.2,136.9, 135.4,133.2,130.8,129.4,128.7,125.2,118.1,116.6,114.2,57.8,45.7,38.2,28.7, 18.5,13.6ppm；HRMS(ESI):m/z,calcd for C₂₂H₁₈F₃N₅O₂S 473.1133(M-H)^-,found 473.1126；IR(KBr):3426,2382,1745,1668,1598,1509,1464,1282,1173,1146,1060, 808cm^-1.

Embodiment 26

The preparation of compound CPU407

Chloroacetic chloride, first synthetic intermediate 20d and 21d, then with identical as compound CPU401 are replaced with ethyl chloroformate Method synthesize compound CPU407, yield 24.0%.m.p.193-195℃；¹H-NMR (300MHz, DMSO-d₆):δ9.73 (s, 1H ,-NH-), 9.19 (s, 1H ,-ArH), 8.74 (s, 1H, Ar-H), 8.02 (d, J=8.4Hz, 1H, Ar-H), 7.46- 7.24 (m, 3H, Ar-H), 5.34 (d, J=17.4Hz, 1H ,-CH-), 4.86-4.81 (m, 1H ,-CH-), 4.69 (d, J= 17.4Hz,1H,-CH-),4.15-4.12(m,2H,-O-CH₂-),3.26-3.23(m,2H,-CH₂-),1.25-1.09(m,3H,- CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ177.7,171.9,153.5,138.1,135.5,135.4,133.2, 130.8,129.5,128.5,125.2,124.5,119.7,117.3,115.6,114.2,60.2,57.8,45.7,28.6, 14.5ppm；HRMS(ESI):m/z,calcd for C₂₁H₁₆F₃N₅O₃S 475.0926(M-H)^-,found 475.0916；IR (KBr):3450,2361,1757,1724,1498,1464,1312,1286,1238,1171,1143,1060cm^-1；HPLC: 94.53%.

Embodiment 27

The preparation of compound CPU408

Chloroacetic chloride, first synthetic intermediate 20e and 21e, then with identical with compound CPU401 are replaced with mesyl chloride Method synthesizes compound CPU408, yield 27.8%.m.p.220-222℃；¹H-NMR(300MHz,DMSO-d₆):δ9.84(s, 1H ,-NH-), 9.19 (d, J=1.6Hz, 1H, Ar-H), 8.74 (d, J=1.6Hz, 1H, Ar-H), 7.32 (d, J=8.3Hz, 1H, Ar-H), 7.20 (s, 1H, Ar-H), 7.3-7.09 (m, 1H, Ar-H), 5.39 (d, J=17.8Hz, 1H ,-CH₂-),4.86- 4.82 (m, H ,-CH-), 4.73 (d, J=17.8Hz, 1H ,-CH₂-),3.28-3.25(m,2H,-CH₂-),3.03(s,3H,- CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ177.7,171.9,171.8,153.5,137.2,135.4,135.3, 133.1,132.3,131.6,130.1,126.2,118.9,117.4,114.2,57.7,57.5,45.6,28.6ppm；HRMS (ESI):m/z,calcd for C₁₉H₁₄F₃N₅O₃S₂ 481.0490(M-H)^-,found 481.0482；IR(KBr):3267, 1753,1618,1488,1461,1380,1314,1268,1149,1059,977,512cm^-1；HPLC:91.94%.

Embodiment 28

The preparation of compound CPU409

Chloroacetic chloride, first synthetic intermediate 20f and 21f, then with identical with compound CPU401 are replaced with fourth sulfonic acid chloride Method synthesizes compound CPU409, yield 29.5%.m.p.245-247℃；¹H-NMR(300MHz,DMSO-d₆):δ9.76(s, 1H ,-NH-), 8.37 (d, J=8.5Hz, 1H ,-ArH), 8.22 (s, 1H ,-ArH), 8.09-8.01 (m, 1H ,-ArH), 7.28 (d, J=7.8Hz, 1H ,-ArH), 7.17-7.12 (m, 1H ,-ArH), 7.02 (d, J=7.8Hz, 1H ,-ArH), 5.34 (d, J= 17.7Hz,1H,-CH-),4.87-4.84(m,2H,-CH₂), 4.50 (dd, J=16.02,16.08Hz, 2H ,-CH₂-),3.29- 3.18(m,2H,-CH₂), 1.62 (t, J=6.9Hz, 2H ,-CH₂-),1.39-1.33(m,2H,-CH₂-),0.88-0.80(m, 3H,-CH₃)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ178.2,172.0,171.0,137.9,136.7,133.7,132.9, 131.7,130.0,126.1,118.3,117.0,116.7,115.0,108.3,57.5,53.5,52.3,50.3,45.5, 43.9,28.6,25.0,20.6,13.7ppm；HRMS(ESI):m/z,calcd for C₂₂H₂₀F₃N₅O₃S₂ 523.0960(M- H)^-,found 523.0961；IR(KBr):3433,2360,2342,1752,1618,1491,1459,1312,1147,405cm^-1；HPLC:93.01%.

Embodiment 29

The preparation (CPU601-CPU611) of Compounds of formula II

The preparation of 2- quinolinecarboxylic acid methyl esters (23)

2- quinolinecarboxylic acid 22 (0.50g, 2.91mmol) is dissolved in 30mL methanol, be slowly added dropwise at 0 DEG C (0.69g, 5.82mmol) thionyl chloride, drop finish, and equality of temperature stirs 30min, are warming up to back flow reaction 8h later, and TLC detects fully reacting, subtracts Concentrated solvent is pressed, is filtered, ethyl acetate filter wash cake obtains off-white powder 0.49g, yield 92.3%.Product directly carries out lower step Reaction.HRMS(ESI):m/z,calcd for C₁₁H₉NO₂ 188.0706(M+H)⁺,found 188.0711。

The preparation of 1,2,3,4- tetrahydroquinoline -2- methyl formate (24)

Intermediate 23 (0.49g, 2.62mmol) is dissolved in 10mL methanol and 20mL tetrahydrofuran mixed solution, and cyano is added Sodium borohydride 0.50g (7.86mmol) adjusts pH to 3-4 with 1M aqueous hydrochloric acid solution after sodium cyanoborohydride dissolution, and maintaining should PH value of solution, normal-temperature reaction 3h, TLC detect fully reacting, solvent are concentrated under reduced pressure, adds 100mL water, with ethyl acetate (30mL × 3) Extraction.Merge organic layer, saturated common salt washing, anhydrous sodium sulfate drying.Filtering, evaporating solvent under reduced pressure obtain yellow oil 0.41g, yield 81.9%.Product directly carries out the next step.HRMS(ESI):m/z,calcd for C₁₁H₁₃NO₂ 192.1019(M+H)⁺,found 192.1021。

The preparation of compound (CPU601)

Intermediate 10 (0.23g, 1.01mmol) is dissolved in 10mL DMF, under room temperature be added intermediate 24 (0.19g, 1.01mmol), equality of temperature stirs 5min, and TLC detects fully reacting, is added in 30mL ice water, ethyl acetate (30mL × 3) extraction, Merge organic layer, saturated common salt washing, anhydrous sodium sulfate drying.Filtering, evaporating solvent under reduced pressure, column chromatograph to obtain white solid 0.20g, yield 51.7%.m.p.218-220℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.43 (d, J=7.3Hz, 1H, Ar- ), H 8.39 (d, J=8.0Hz, 1H, Ar-H), 8.26 (s, 1H, Ar-H), 8.04 (d, J=7.1Hz, 1H, Ar-H), 7.22- 7.31(m,3H,Ar-H),4.81-4.84(m,1H,-CH-),3.05-3.16(m,2H,-CH₂-),2.39-2.50(m,2H,- CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ176.8,171.0,139.1,136.3,134.4,132.4,130.6, 128.4,128.3,123.2,122.5,119.6,118.2,115.0,108.7,60.5,25.6,23.8ppm；HRMS(ESI): m/z,calcd for C₁₉H₁₂F₃N₃OS 386.0580(M-H)^-,found 386.0587；IR(KBr):3420,3124, 1758,1491,1417,1410,1313,1260,1132,1056,804,758cm^-1.

Embodiment 30

The preparation of compound (CPU602)

N- acetyl group -1,2, the preparation of 3,4- tetrahydroquinoline -2- methyl formates (25)

Intermediate 24 (0.38g, 2.10mmol) is dissolved in 20mL methylene chloride, and (2.56g, 2.52mmol) is added at 0 DEG C Chloroacetic chloride (0.24g, 3.16mmol) is added after stirring 5min in triethylamine, equality of temperature, is warming up to 30 DEG C of reaction 1h, TLC detection reactions Completely, 100mL water is added in reaction solution, and methylene chloride (30mL × 3) extraction merges organic layer, saturated common salt washing, anhydrous slufuric acid Sodium is dry.Filtering, evaporating solvent under reduced pressure, column chromatograph to obtain 0.38g faint yellow solid, yield 77.7%.Product directly carries out lower step Reaction.HRMS(ESI):m/z,calcd for C₁₃H₁₅NO₃ 234.1125(M+H)⁺,found 234.1132.

The preparation of bromo- 1,2,3,4- tetrahydroquinoline -2- methyl formate (26) of N- acetyl group -6-

Intermediate 25 (0.38g, 1.63mmol) is dissolved in 10mL anhydrous DMF, be slowly added dropwise at 0 DEG C (0.26g, 1.63mmol) bromine, equality of temperature stir 40min, and TLC detects fully reacting, and reaction solution is passed through nitrogen discharge excess liquid bromine, adds 100mL water, ethyl acetate (30mL × 3) extraction merge organic layer, saturated common salt washing, anhydrous sodium sulfate drying.Filtering, subtracts Solvent is evaporated off in pressure, and column chromatographs to obtain 0.35g orange solids, yield 68.8%.Product directly carries out the next step.HRMS(ESI):m/ z,calcd for C₁₃H₁₄BrNO₃ 312.0230(M+H)⁺,found 312.0235.

The preparation of bromo- 1,2,3,4- tetrahydroquinoline -2- formic acid (27) of 6-

0.32g (1.12mmol) intermediate 26 is dissolved in 20ml concentrated hydrochloric acid, is warming up to reflux, reacts 2h, TLC detection reaction Completely, evaporating solvent under reduced pressure obtains brown solid 0.27g, yield 94.5%.Product directly carries out the next step.HRMS(ESI): m/z,calcd for C₁₀H₁₀BrNO₂ 255.9968(M+H)⁺,found 255.9981.

The preparation of bromo- 1,2,3,4- tetrahydroquinoline -2- methyl formate (28) of 6-

Intermediate 27 (0.27g, 1.05mmol) is dissolved in 30mL methanol, be slowly added dropwise at 0 DEG C (0.25g, 2.10mmol) thionyl chloride, drop finish, and equality of temperature stirs 30min, are warming up to back flow reaction 6h later, and TLC detects fully reacting, subtracts Concentrated solvent is pressed, filters, obtains orange solids 0.27g, yield 96.2%.Product directly carries out the next step.HRMS(ESI):m/ z,calcd for C₁₁H₁₂NO₂ 270.0124(M+H)⁺,found 270.0151.

The preparation of compound (CPU602)

Intermediate 10 (0.23g, 1.01mmol) is dissolved in 10mL DMF, and it is intermediate that (0.27g, 1.01mmol) is added under room temperature Body 28, equality of temperature stir 5min, and TLC detects fully reacting, are added in 30mL ice water, and ethyl acetate (30mL × 3) extraction is associated with Machine layer, saturated common salt washing, anhydrous sodium sulfate are dry.Filtering, evaporating solvent under reduced pressure, column chromatograph to obtain white solid 0.22g, yield 47.2%.m.p.219-222℃；¹H-NMR(300MHz,DMSO-d₆): δ 8.84 (d, J=8.9Hz, 1H, Ar-H), 8.40 (d, J =8.2Hz, 1H, Ar-H), 8.26 (s, 1H, Ar-H), 8.04 (d, J=7.4Hz, 1H, Ar-H), 7.57-7.49 (m, 2H, Ar- ), H 4.83 (dd, J=11.5Hz, 3.6Hz, 1H ,-CH-), 3.16-3.11 (m, 2H ,-CH₂-),2.33-2.28(m,2H,- CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ176.9,170.8,138.1,136.3,134.4,131.9,131.4, 130.9,128.3,128.2,127.5,123.2,120.3,117.8,115.0,108.7,60.4,25.3,23.4ppm；HRMS (ESI):m/z,calcd for C₁₉H₁₂BrF₃N₃OS 463.9686(M-H)^-,found 463.9676；IR(KBr):3447, 3133,1754,1440,1401,1317,1258,1144,1084,749,544cm^-1.

Embodiment 31

The preparation of compound (CPU603)

The preparation of N- oxidation -6- oxyquinoline (30)

Intermediate 29 (2.00g, 13.79mmol) is dissolved in 50mL ethyl acetate, be added in three times at 0 DEG C (5.95g, 34.48mmol) metachloroperbenzoic acid, N₂Protection, equality of temperature react 2h.It is to slowly warm up to room temperature the reaction was continued 6h, TLC detection is anti- Brown solid 1.86g, yield 83.8% should be filtered to obtain completely.Product directly carries out the next step.¹H-NMR(300MHz, DMSO-d₆): δ 10.67 (s, 1H ,-OH), 8.42 (d, J=8.8Hz, 1H, Ar-H), 8.01 (d, J=9.2Hz, 1H, Ar-H), 7.90 (d, J=8.4Hz, 1H, Ar-H), 7.50 (dd, J=9.0Hz, 2.6Hz, 1H, Ar-H), 10.67 (s, 1H, Ar-H) ppm.

The preparation of -2 itrile group quinoline (31) of 6- oxyquinoline

Intermediate 30 (1.86g, 11.55mmol) is dissolved in 50mL acetonitrile, is added triethylamine (5.36g, 53.13mmol), three Methyl silicon cyanogen (4.92g, 49.66mmol), normal-temperature reaction 10h.TLC detects fully reacting, and evaporating solvent under reduced pressure obtains brown oil Object 1.87g.Yield 95.1%.Product directly carries out the next step.HRMS(ESI):m/z,calcd for C₁₀H₆N₂O 171.0553(M+H)⁺,found 171.0568.

The preparation of 6- oxyquinoline -2- formic acid (32)

Intermediate 31 (1.87g, 10.99mmol) is dissolved in 30mL concentrated hydrochloric acid, is warming up to back flow reaction 7h.TLC detection reaction Completely, evaporating solvent under reduced pressure obtains brown solid 1.73g.Yield 83.3%.Product directly carries out the next step.HRMS(ESI): m/z,calcd for C₁₀H₆N₂O 190.0499(M+H)⁺,found 190.0518.

The preparation of 6- oxyquinoline -2- methyl formate (33)

Intermediate 32 (1.73g, 9.10mmol) is dissolved in 30mL methanol, be slowly added dropwise at 0 DEG C (2.17g, 18.21mmol) thionyl chloride, drop finish, and equality of temperature stirs 30min, are warming up to back flow reaction 12h later, and TLC detects fully reacting, Evaporating solvent under reduced pressure obtains brown oil 1.68g, yield 91.9%.Product directly carries out the next step.HRMS(ESI):m/z, calcd for C₁₁H₉NO₃ 204.0655(M+H)⁺,found 204.0651.

6- hydroxyl -1,2, the preparation of 3,4- tetrahydroquinoline -2- methyl formates (34)

Intermediate 33 (1.68g, 8.27mmol) is dissolved in 15mL methanol and 30mL tetrahydrofuran mixed solution, and cyano is added Sodium borohydride (1.56g, 24.83mmol) adjusts pH to 3-4 with 1M aqueous hydrochloric acid solution after sodium cyanoborohydride dissolution, maintains The pH value of solution, normal-temperature reaction for 24 hours, TLC detect fully reacting, be concentrated under reduced pressure solvent, add 100mL water, with ethyl acetate (30mL × 3) it extracts.Merge organic layer, saturated common salt washing, anhydrous sodium sulfate drying.Filtering, evaporating solvent under reduced pressure, column chromatograph to obtain yellow Grease 0.76g, yield 37.4%.HRMS(ESI):m/z,calcd for C₁₁H₁₃NO₃ 208.0968(M+H)⁺,found 208.0962.

The preparation of compound (CPU603)

Intermediate 10 (0.23g, 1.01mmol) is dissolved in 10mL DMF, and it is intermediate that (0.21g, 1.01mmol) is added under room temperature Body 34, equality of temperature stir 5min, and TLC detects fully reacting, are added in 30mL ice water, and ethyl acetate (30mL × 3) extraction is associated with Machine layer, saturated common salt washing, anhydrous sodium sulfate are dry.Filtering, evaporating solvent under reduced pressure, column chromatograph to obtain white solid 0.10g, yield 24.8%.m.p.221-223℃；¹H-NMR(300MHz,DMSO-d₆): δ 9.60 (s, 1H ,-OH), 8.63 (d, J=9.6Hz, 1H, Ar-H), 8.39 (d, J=8.2Hz, 1H, Ar-H), 8.25 (s, 1H, Ar-H), 8.03 (d, J=8.2Hz, 1H, Ar-H), 6.70-6.68 (m, 2H, Ar-H), 4.75 (dd, J=11.4Hz, 4.0Hz, 1H ,-CH-), 3.08-2.95 (m, 2H ,-CH₂-), 2.39-2.26(m,2H,-CH₂-)ppm；¹³C-NMR(75MHz,DMSO-d₆):δ176.9,171.0,155.5,139.1, 136.3,134.4,132.6,130.6,128.4,128.3,123.2,122.5,119.6,118.2,115.0,108.7,60.5, 25.6,23.9ppm；HRMS(ESI):m/z,calcd for C₁₉H₁₂F₃N₃O₂S 402.0530(M-H)^-,found 402.0660；IR(KBr):3373,3128,1773,1473,1409,1354,1270,1168,1132,1099,512cm^-1.

Claims

1. the compound or its pharmaceutically acceptable salt of general formula I or II:

Wherein:

X represents carbon atom or nitrogen-atoms；

R₁Represent halogen or C₁~₄Halogenated alkyl；

R₂Represent hydrogen, halogen, cyano or hydroxyl；

R₂Also represent C₁~₄Alkyl, C₁~₄Alkoxy, C₁~₄Halogenated alkyl, cyclopropyl, imidazole radicals, pyrazolyl, triazolyl, tetrazolium Base, morpholinyl, tetrahydrofuran base, piperidyl, piperazinyl, oxetanyl ,-C (O) R^a、-NR^aR^a、-COOR^a、-C(O) NR^aR^b、-C(S)NR^aR^b、-NHC(O)R^a、-NHSO₂R^aOr-SO₂NR^aR^b；Above-mentioned substituent group can be also optionally substituted by a hydroxyl group；Wherein R^a、R^b Respectively independently represent hydrogen, C₁~₃Alkyl, C₁~₃Hydroxyalkyl or R^a、R^bConnection formed nafoxidine base, piperidyl, morpholine base, Piperazinyl or N-methyl piperazinyl；

R₃Represent hydrogen or halogen；

R₄Represent hydrogen, halogen, cyano, hydroxyl or C₁~₄Alkoxy.

2. the compound of claim 1 or its pharmaceutically acceptable salt, wherein R₁Represent halogen or trifluoromethyl.

3. the compound of claim 1 or its pharmaceutically acceptable salt, wherein R₂Represent hydrogen, halogen, cyano, hydroxyl, cyclopropyl Base ,-NHC (O) R^a、-NHSO₂R^aOr-SO₂NR^aR^b；Wherein R^a、R^bRespectively independently represent hydrogen, C₁~₂Alkyl, C₁~₂Hydroxyalkyl or R^a、R^bConnection forms nafoxidine base, piperidyl, morpholine base, piperazinyl or N-methyl piperazinyl.

4. the compound of claim 1 or its pharmaceutically acceptable salt are the compound of following any structure:

5. a kind of pharmaceutical composition, wherein containing the compound of any one of claims 1 to 4 or its pharmaceutically acceptable salt And pharmaceutically acceptable carrier.

6. the compound of any one of claims 1 to 4 or its pharmaceutically acceptable salt, which are used as, prepares the competing of androgen receptor The purposes of striving property antagonist pharmaceuticals.

7. the compound of any one of claims 1 to 4 or its pharmaceutically acceptable salt are used to prepare treatment malignant tumour The purposes of drug.