CN106660974B - 1,2,5- furodiazole derivative, preparation method containing sulfamoyl and its application in medicine - Google Patents

Abstract

A kind of 1,2,5- furodiazole derivative containing sulfamoyl, preparation method and its application in medicine.Specifically; as lead to formula (I) shown in containing sulfamoyl 1; 2; 5- furodiazole derivative; preparation method and pharmaceutical composition containing the derivative and its as IDO inhibitor; it can be used for treating the disease for the tryptophan metabolic pathway pathological characteristics that there is IDO to mediate, the disease includes tumour, cancer, Alzheimer disease, autoimmune disease, depression, anxiety disorder, cataract, mental handicape and AIDS.Each substituent group of its formula of (I) is identical as the definition in specification.

Description

It 1 containing sulfamoyl, 2,5- furodiazole derivative, preparation methods and its is curing Application on medicine

Technical field

The invention belongs to field of medicaments, are related to a kind of 1 containing sulfamoyl, the synthesis side of 2,5- furodiazole derivatives Method and its application in medicine.The invention discloses it as IDO inhibitor, for treating the tryptophan mediated with IDO The disease of metabolic pathway pathological characteristics, the disease include cancer, Alzheimer disease, autoimmune disease, depression Disease, anxiety disorder, cataract, mental handicape, AIDS.

Background technique

Tumour is to seriously endanger one of the major disease of human life, more than half occurs in developing country.It dislikes in China Property tumor incidence it is totally in rising trend, disease incidence is with the speed increase of average annual 3%-5%, it is contemplated that arrive the year two thousand twenty, China will There is the raw cancer of 4,000,000 human hairs, 3,000,000 people die of cancer, the main reason is that: aging, urbanization, industrialization and living habit Change.In Chinese Hospitals medication market, the steady-state growth always in recent years of the marketing scale of anti-tumor drug reaches for 2012 664.2 hundred million yuan, increase by 13.07% on a year-on-year basis, it is contemplated that by 2017, the market scale of anti-tumor drug was up to 1055.7 hundred million Member increases by 7.57% on a year-on-year basis.

Since malignant tumour is without limitation growth and infiltration, transfer, clinical three big conventional treatments (hands used now Art, radiation and chemotherapy) tumour cell can not be cut off or thoroughly killed completely, therefore often there is metastases or recurrence.Tumour is raw Object treatment is the new treatment that treatment and prevention of tumour is carried out using modern biotechnology and its Related product, because of it safely, effectively, it is bad anti- The features such as low is answered, the 4th kind of mode of the oncotherapy after operation, radiotherapy, chemotherapy is become, by transferring the natural of host Defense mechanism (such as the tumor immune escape mechanism for inhibiting IDO to mediate) gives the very strong substance of naturally-produced targeting Obtain antitumor effect.

Indoles amine-pyrroles -2,3- dioxygenase (Indoleamine-pyrrole-2,3-dioxygenase, IDO) is one Kind iron content ferroheme monomeric protein, is made of 403 amino acid residues, the α-helixstructure domain folded including two, big structure Domain includes catalytic pocket, and with IDO can occur for substrate the effects of hydrophobic in catalytic pocket.IDO is that catalysis tryptophan transfer turns to first The enzyme of acyl kynurenin is widely distributed in people and other mammals (rabbit, mouse) in the tissue in addition to liver, is other than liver It can be uniquely catalyzed the rate-limiting enzyme of tryptophan catabolism, and tryptophan is amino acid necessary to cell maintains activation and is proliferated, It is also to constitute the indispensable important component of protein.IDO and interferon (interferon, IFN), interleukins The cytokine profiles such as (interleukin, IL), tumor necrosis factor are in close relations, they can be activated under certain condition IDO.And there are the horizontal very sensitive point of adjustment of a tryptophan in the cell cycle of T- cell, on the one hand, IDO makes part Tryptophan depletion causes T- cells arrest in the G1 interim phase, to inhibit the proliferation of T cell；On the other hand, IDO is catalyzed color The primary product cynruin that propylhomoserin metabolism generates is caused Cellular Oxidation agent and antioxidant to change by Mediated by Free Radicals and is lured T- Apoptosis is led, this is the intrinsic immunosuppression mechanism for being present in body.At present a large number of studies show that IDO is thin in leukaemia Higher expression in born of the same parents, keeps local T cell proliferation suppressed, and the immune response for inhibiting T- cell-mediated turns T- cell activation signal It leads and is obstructed, thus the attack of mediate tumor cell escape immune system.It has been found that most of mankind's tumor groups are expressed with becoming second nature IDO.Therefore, IDO is the target of the cancer immunotherapy of a tool potentiality.

1- methyl tryptophan (1-methyltryptophan) is small by taking orally for NewLink Genetics company exploitation Molecule IDO inhibitor enters the clinical second phase in the U.S. at present for treating metastatic breast cancer and solid tumor.Research shows that 1- Methyl tryptophan can enhance tumour cell to the sensibility of the immunostimulation of T- cell in vitro, energy in animal model in vivo Delay the growth of tumour cell and the antitumous effect of Reinforcement chemotherapy drug, and almost all spontaneous tumors are acted as With.In addition, INCB-24360 is also carrying out clinic in a series of oral IDO micromolecular inhibitors that Incyte company is researching and developing Second phase trial is mainly used for treating the kinds cancer including myelodysplastic syndrome.

The inhibitor patent application of disclosed selective depression IDO include WO2004094409, WO2006122150, WO2007075598, WO2010005958 and WO2014066834 etc..

IDO inhibitor has a good application prospect as drug in pharmaceuticals industry, but not yet finds at present well IDO inhibitor can be used as marketed drug, in order to achieve the purpose that better oncotherapy effect, better meet the market demand, It is desirable that the selective IDO inhibitor of the high-efficiency low-toxicity of a new generation can be developed.The present invention will provide a kind of new structural Selective IDO inhibitor, and the compound for being found to have this class formation shows excellent effect and effect, it is especially excellent In medicine generation, absorbs activity.

Summary of the invention

The purpose of the present invention is to provide a kind of logical formula (I) compound represented or its tautomers, mesomer, outer Raceme, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt, shown in formula of (I) Compound structure it is as follows:

Wherein:

Mixture selected from cis-isomer, transisomer or cis-trans-isomer；

R¹Selected from alkyl, naphthenic base and alkoxy, wherein the alkyl, naphthenic base, alkoxy are optional each independently Further replaced one or more substituent groups in hydroxyl, alkyl, alkoxy and halogenated alkyl；

R²Selected from hydrogen atom, alkyl, naphthenic base and alkoxy, wherein the alkyl, naphthenic base, alkoxy are respectively independent Ground is optionally further replaced one or more substituent groups in hydroxyl, alkyl, alkoxy and halogenated alkyl；

Or R¹And R²Carbon atom in connection is formed together naphthenic base or heterocycle, wherein the naphthenic base, heterocycle Base is optionally further selected from alkyl, halogen, cyano, naphthenic base, heterocycle, aryl, heteroaryl ,-NR⁶R⁷、-C(O)NR⁶R⁷、- NHS(O)_mR⁸、-S(O)_pR⁸、-NHC(O)R⁸、-OR⁸、-C(O)R⁸、-OC(O)R⁸With-C (O) OR⁸In one or more substituent groups It is replaced；

R³And R⁴It is each independently selected from hydrogen atom, alkyl, cyano, amino, halogen, alkenyl, alkynyl, hydroxyl, nitro, ring Alkyl, heterocycle, aryl and heteroaryl, wherein alkyl, naphthenic base, heterocycle, aryl or the heteroaryl are each independently Optionally further by hydroxyl, halogen, amino, cyano, alkyl, alkoxy, naphthenic base, heterocycle, aryl and heteroaryl One or more substituent groups replaced；

R⁵Selected from alkyl and amino, the alkyl and amino optionally further by selected from hydroxyl, halogen, amino, cyano, Replaced one or more substituent groups in alkyl, alkoxy, naphthenic base, heterocycle, aryl and heteroaryl；The alkyl is preferred C_1~4Alkyl；

R⁶And R⁷It is each independently selected from hydrogen atom, alkyl, naphthenic base, heterocycle, aryl and heteroaryl, wherein described Alkyl, naphthenic base, heterocycle, aryl or heteroaryl are optionally further selected from hydroxyl, halogen, alkyl, alcoxyl each independently Replaced one or more substituent groups in base, naphthenic base, heterocycle, aryl and heteroaryl；

R⁸Selected from hydrogen atom, alkyl, amino, naphthenic base, heterocycle, aryl and heteroaryl, wherein the alkyl, cycloalkanes Base, heterocycle, aryl or heteroaryl are optionally further selected from alkyl, halogen, hydroxyl, amino, cyano, alcoxyl each independently Replaced one or more substituent groups in base, naphthenic base, heterocycle, aryl and heteroaryl；

M is 0,1,2；

N is 0,1,2；And

P is 0,1,2.

In a preferred embodiment of the present invention, lead to formula (I) compound represented or its tautomer, meso Body, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt, wherein R³And R⁴Respectively Independently selected from hydrogen atom, halogen, alkyl or halogenated alkyl, the halogen is preferably fluorine, chlorine or bromine, and the alkyl is preferably C_1~4Alkyl, the halogenated alkyl are preferably trifluoromethyl.

In a preferred embodiment of the present invention, lead to formula (I) compound represented or its tautomer, meso Body, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt, for logical formula (II) Compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer or its mixing Object form or its officinal salt:

Wherein:

R¹For alkyl or alkoxy, the alkyl or alkoxy are further taken by one or more hydroxyls or alkoxy Generation；Wherein preferred R¹For the alkyl replaced by hydroxyl or alkoxy；

R³~R⁵, m, n be as defined in logical formula (I).

Currently preferred logical formula (II) compound represented or its tautomer, mesomer, racemic modification, mapping Isomers, diastereoisomer, or mixtures thereof form or its officinal salt:

Wherein:

R¹For alkyl or alkoxy, the alkyl or alkoxy are further replaced by one or more hydroxyls or alkoxy；

R³For halogen, alkyl or halogenated alkyl, more preferable halogen；

R⁴For halogen, alkyl or halogenated alkyl, more preferable halogen；

R⁵For amino；

The integer that m is 0~2, more preferably 0；

The integer that n is 0~2, more preferably 1.

In a preferred embodiment in accordance with this invention, lead to formula (I) compound represented or its tautomer, interior disappear Revolve body, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt, be general formula (III) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer or Its form of mixtures or its officinal salt:

Wherein:

A is 3~8 member rings, preferably 3~6 member rings, wherein the ring is selected from carbocyclic ring or heterocycle；

R³~R⁵, m, n be as defined in logical formula (I).

It is currently preferred logical formula (III) compound represented or its tautomer, mesomer, racemic modification, right Reflect isomers, diastereoisomer, or mixtures thereof form or its officinal salt:

Wherein:

A is naphthenic base, more preferable cyclopropyl；

R³For halogen, alkyl or halogenated alkyl, more preferable halogen；

R⁴For halogen, alkyl or halogenated alkyl, more preferable halogen；

R⁵For amino；

The integer that m is 0~2；

The integer that n is 1~2.

It prepares logical formula (I) compound represented or its tautomer the present invention also provides a kind of, mesomer, outside disappear Revolve body, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt method, this method comprises:

Open loop obtains logical formula (I) compound to logical formula (IV) compound under alkaline condition, wherein the alkaline condition Reagent is potassium carbonate or sodium hydroxide；

Wherein:

R¹~R⁵, m, n be as defined in logical formula (I).

The invention further relates to a kind of logical formula (IV) compound represented or its tautomer, mesomer, racemic modification, Enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt:

Wherein:

R¹~R⁵, m, n be as defined in logical formula (I).

In a preferred embodiment in accordance with this invention, a kind of logical formula (IV) compound represented or its tautomerism Body, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt, be Logical formula (V) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, Or mixtures thereof form or its officinal salt:

Wherein:

R¹、R³~R⁵, m, n be as defined in logical formula (I).

In a preferred embodiment in accordance with this invention, a kind of logical formula (IV) compound represented or its tautomerism Body, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt, be Logical formula (VI) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, Or mixtures thereof form or its officinal salt:

Wherein:

A is 3~8 member rings, wherein the ring is selected from carbocyclic ring or heterocycle；

R³~R⁵, m, n be as defined in logical formula (I).

The present invention by a series of reaction, synthesized typical logical formula (I), (II), (III) compound include, but It is not limited to:

Another aspect of the present invention relates to a kind of pharmaceutical compositions, logical formula (I), (II) containing treatment effective dose and (III) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer or Its form of mixtures or pharmaceutical salt and one or more pharmaceutically acceptable carriers, diluent or excipient.

The invention further relates to a kind of methods for preparing above-mentioned composition comprising will be shown in logical formula (I), (II) and (III) Compound or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof shape Formula or its pharmaceutical salt are mixed with pharmaceutically acceptable carrier, diluent or excipient.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, Or mixtures thereof enantiomter, diastereoisomer form or its officinal salt, or prepared comprising its pharmaceutical composition For prevent and/or Prevention have IDO mediate tryptophan metabolic pathway pathological characteristics disease drug in Purposes.IDO inhibitor can be used for the inhibition of cardiac disorder and treat other with the IDO tryptophan metabolic pathway mediated The disease of pathological characteristics, these diseases include the infection of the virus such as AIDS, and Lyme disease and streptococcal infection etc. are thin Born of the same parents' infection, Neurodegenerative conditions (such as Alzheimer disease, Huntington disease and the gloomy disease of bat gold), autoimmune disease, suppression Strongly fragrant disease, anxiety disorder, cataract, mental handicape, AIDS, cancer (including T cell leukaemia and colon cancer), ocular disease state (such as cataract and age-related yellow) and autoimmune disease, wherein the cancer can be selected from mammary gland Cancer, cervical carcinoma, colon cancer, lung cancer, gastric cancer, the carcinoma of the rectum, cancer of pancreas, the cancer of the brain, cutaneum carcinoma, carcinoma of mouth, prostate cancer, osteocarcinoma, kidney Cancer, oophoroma, bladder cancer, liver cancer, fallopian tube cneoplasms, ovarioncus, peritoneal tumor, IV phase melanoma, solid tumor, neuroglia The renal tumor of tumor, spongioblastoma, hepatocellular carcinoma, mastoid process, head and neck neoplasm, leukaemia, lymthoma, myeloma and non-small thin Born of the same parents' lung cancer.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, mappings Or mixtures thereof isomers, diastereoisomer form or its officinal salt, or comprising its pharmaceutical composition, be used for pre- The disease of the pathological characteristics for the tryptophan metabolic pathway that there is anti-and/or Prevention IDO to mediate.These diseases include such as The infection of the virus such as AIDS, the cell infections such as Lyme disease and streptococcal infection, Neurodegenerative conditions (such as alzheimer ' Silent disease, Huntington disease and clap the gloomy disease of gold), autoimmune disease, depression, anxiety disorder, cataract, mental handicape, AIDS, Cancer (including T cell leukaemia and colon cancer), ocular disease state (such as cataract and age-related yellow) and Autoimmune disease, wherein the cancer can be selected from breast cancer, cervical carcinoma, colon cancer, lung cancer, gastric cancer, the carcinoma of the rectum, pancreas Gland cancer, the cancer of the brain, cutaneum carcinoma, carcinoma of mouth, prostate cancer, osteocarcinoma, kidney, oophoroma, bladder cancer, liver cancer, fallopian tube cneoplasms, ovary Tumor, peritoneal tumor, IV phase melanoma, solid tumor, glioma, spongioblastoma, hepatocellular carcinoma, mastoid process are renal Tumor, head and neck neoplasm, leukaemia, lymthoma, myeloma and non-small cell lung cancer.

The invention further relates to a kind of diseases for the tryptophan metabolic pathway that there is IDO to mediate for Prevention and/or Prevention The method of the disease of feature of science comprising to patient apply treatment effective dose logical formula (I) compound represented or its mutually Or mixtures thereof tautomeric, mesomer, racemic modification, enantiomter, diastereoisomer form or its is pharmaceutically acceptable Salt, or the pharmaceutical composition comprising it.These diseases include the infection of the virus such as AIDS, such as Lyme disease and streptococcus sense The cell infections such as dye, Neurodegenerative conditions (such as Alzheimer disease, Huntington disease and the gloomy disease of bat gold), autoimmune disease Disease, depression, anxiety disorder, cataract, mental handicape, AIDS, cancer (including T cell leukaemia and colon cancer), eyes disease Diseased state (such as cataract and age-related yellow) and autoimmune disease, wherein the cancer can be selected from Breast cancer, cervical carcinoma, colon cancer, lung cancer, gastric cancer, the carcinoma of the rectum, cancer of pancreas, the cancer of the brain, cutaneum carcinoma, carcinoma of mouth, prostate cancer, bone Cancer, kidney, oophoroma, bladder cancer, liver cancer, fallopian tube cneoplasms, ovarioncus, peritoneal tumor, IV phase melanoma, solid tumor, mind Through glioma, spongioblastoma, hepatocellular carcinoma, the renal tumor of mastoid process, head and neck neoplasm, leukaemia, lymthoma, myeloma and Non-small cell lung cancer.

Another aspect of the present invention is related to a kind of method for the treatment of cancer, and this method includes applying treatment effective dose to patient Logical formula (I) of the invention described in compound or its tautomer, mesomer, racemic modification, enantiomter, non-right Reflect or mixtures thereof isomers form or its officinal salt.This method shows curative effect outstanding and less side effect, wherein The cancer can be selected from breast cancer, cervical carcinoma, colon cancer, lung cancer, gastric cancer, the carcinoma of the rectum, cancer of pancreas, the cancer of the brain, cutaneum carcinoma, mouth Chamber cancer, prostate cancer, osteocarcinoma, kidney, oophoroma, bladder cancer, liver cancer, fallopian tube cneoplasms, ovarioncus, peritoneal tumor, IV phase are black The renal tumor of melanoma, solid tumor, glioma, spongioblastoma, hepatocellular carcinoma, mastoid process, head and neck neoplasm, white blood Disease, lymthoma, myeloma and non-small cell lung cancer, preferably fallopian tube cneoplasms, peritoneal tumor, IV phase melanoma, myeloma And breast cancer, more preferably breast cancer.

Pharmaceutical composition containing active constituent, which can be, is suitable for oral form, such as tablet, dragee, pastille, water Or oil suspension, dispersible powder or particle, lotion, hard or soft capsule or syrup or elixir.It can be any according to this field Know that the method for preparing Pharmaceutical composition prepares Orally administered composition, such composition can containing it is one or more it is selected from the following at Point: sweetener, corrigent, colorant and preservative, to provide pleasing and palatable pharmaceutical formulation.Tablet contain active constituent and The suitable nontoxic pharmaceutical excipient for preparing tablet for mixing.These excipient can be inert excipient, such as carbon Sour calcium, sodium carbonate, lactose, calcium phosphate or sodium phosphate；Granulating agent and disintegrating agent, such as microcrystalline cellulose, cross-linked carboxymethyl fiber Plain sodium, cornstarch or alginic acid；Adhesive, such as starch, gelatin, polyvinylpyrrolidone or Arabic gum and lubricant, example Such as magnesium stearate, stearic acid or talcum powder.These tablets can not be coated or can be by covering the taste of drug or in gastrointestinal tract Middle delay disintegration and absorption, thus the known technology for providing slow releasing function in a long time is coated.For example, water can be used Dissolubility taste masked substance, such as hydroxypropyl methyl cellulose or hydroxypropyl cellulose, or extend time substance such as ethyl fibre Dimension element, acetylbutyrylcellulose.

Also available wherein active constituent mixes hard bright with inert solid diluent such as calcium carbonate, calcium phosphate or kaolin Glue capsule or in which active constituent and water-solubility carrier such as polyethylene glycol or oily solvent such as peanut oil, atoleine or olive The Perle of olive oil mixing provides oral preparation.

Water slurry contains active material and the suitable excipient for preparing water slurry for mixing.Such excipient is Suspending agent, such as sodium carboxy methyl cellulose, methylcellulose, hydroxypropyl methyl cellulose, mosanom, polyvinylpyrrolidone And Arabic gum；Dispersing agent or wetting agent can be the contracting of naturally-produced phosphatide such as lecithin or alkylene oxide and fatty acid Close the condensation product of product such as Myrj 45 or ethylene oxide and long-chain fatty alcohol, such as 17 carbon ethylidene Oxygroup cetanol (heptadecaethyleneoxy cetanol) or ethylene oxide and the portion as derived from fatty acid and hexitol The condensation product of ester, such as polyoxyethylene sorbitol monoleate or ethylene oxide is divided to spread out with by fatty acid and hexitan The condensation product of raw partial ester, such as polyethylene oxide Sorbitan Monooleate.Aqueous suspension can also containing a kind of or Determination of Preservatives such as ethylparaben or nipalgin n-propyl, one or more colorants, one or more corrigents and one Kind or a variety of sweeteners, such as sucrose, saccharin or aspartame.

Oil suspension can by making active constituent be suspended in vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or It is formulated in mineral oil such as atoleine.Oil suspension can contain thickener, such as beeswax, hard paraffin or cetanol.It can Above-mentioned sweetener and corrigent is added, to provide palatable preparation.It can be by the way that antioxidant such as Butylated Hydroxyanisole or α-be added Give birth to these compositions of phenol preservation.

It can make to be suitable for preparing that water is suspended dispersible powder also and particle provides active constituent and is used for by the way that water is added Mixed dispersing agent or wetting agent, suspending agent or one or more preservatives.Suitable dispersing agent or wetting agent and suspending agent can Illustrate above-mentioned example.Other excipient such as sweetener, corrigent and colorant can also be added.By the way that antioxidant example is added As ascorbic acid saves these compositions.

Pharmaceutical composition of the invention is also possible to the form of oil in water emulsion.Oil mutually can be vegetable oil such as olive oil Or or mixtures thereof peanut oil or mineral oil such as atoleine.Suitable emulsifier can be naturally-produced phosphatide, such as Soybean lecithin and the ester as derived from fatty acid and hexitan or partial ester such as sorbitan monooleate and the partial ester and ring The condensation product of oxidative ethane, such as polyoxyethylene sorbitol monoleate.Emulsion can also contain sweetener, corrigent, prevent Rotten agent and antioxidant.Syrup and elixir can be prepared with Sweetening agents such as glycerol, propylene glycol, sorbierite or sucrose.Such preparation Moderator, preservative, colorant and antioxidant can be contained.

Pharmaceutical composition can be sterile injectable aqueous form.Can have in the acceptable solvent and solvent used Water, ringer's solution and isotonic sodium chlorrde solution.Aseptic injection preparation can be the aseptic injection that wherein active constituent is dissolved in oily phase Oil-in-water microemulsion.Such as active constituent is dissolved in the mixture of soybean oil and lecithin.Then water and sweet is added in oil solution Processing forms micro emulsion in the mixture of oil.Can be by a large amount of injections in part, it will be in injection or the blood flow of micro emulsion injection patient.Or Person preferably gives solution and micro emulsion in the way of it can keep the compounds of this invention constant circulating concentration.To keep this constant dense Degree, can be used continuous intravenous delivery device.The example of this device is Deltec CADD-PLUS.TM.5400 type vein note Penetrate pump.

Pharmaceutical composition can be for intramuscular and the aseptic injection water of subcutaneous administration or the form of oil suspension.It can be by Know technology, the dispersing agent or wetting agent and suspending agent for being suitable for those described above prepare the suspension.Aseptic injection preparation can also be with It is the aseptic injectable solution or suspension prepared in the acceptable diluent of nontoxic parenteral or solvent, such as 1,3-BDO The solution of middle preparation.Furthermore, it is convenient to use sterile fixed oil as solvent or suspension media.For this purpose, usable include Any reconciliation fixing oil including synthetic glycerine list or diester.In addition, fatty acid such as oleic acid can also prepare injection.

The compounds of this invention can be given by the suppository form for rectally.It can be by by drug and at normal temperatures For solid but in the rectum it is liquid, thus can dissolves and discharge the suitable nonirritant excipient mixing of drug in the rectum To prepare these pharmaceutical compositions.Substance of this kind includes the poly- second of cocoa butter, glycerin gelatine, hydrogenated vegetable oil, various molecular weight The mixture of the aliphatic ester of two pure and mild polyethylene glycol.

Well-known to those skilled in the art, the dosage of drug depends on many factors, including but is not limited to Following factor: the activity of particular compound used, the age of patient, the weight of patient, the health status of patient, patient row Quilt, the diet of patient, administration time, administration mode, the rate of excretion, the combination of drug etc.；In addition, optimal therapeutic modality is such as The type of the mode for the treatment of, the consumption per day of general formula compound or pharmaceutical salt can be verified according to traditional therapeutic scheme.

Detailed description of the invention

Unless stated to the contrary, the term used in the specification and in the claims has following meanings.

Term " alkyl " refers to saturated aliphatic hydrocarbons group, is the linear chain or branched chain group comprising 1 to 20 carbon atom, excellent The alkyl containing 1 to 12 carbon atom is selected, more there is alkyl of the choosing containing 1 to 6 carbon atom.Non-limiting example include methyl, Ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl, sec-butyl, n-pentyl, 1,1- dimethyl propyl, 1,2- diformazan Base propyl, 2,2- dimethyl propyl, 1- ethyl propyl, 2- methyl butyl, 3- methyl butyl, n-hexyl, 1- Ethyl-2-Methyl third Base, 1,1,2- thmethylpropyl, 1,1- dimethylbutyl, 1,2- dimethylbutyl, 2,2- dimethylbutyl, 1,3- dimethyl butyrate Base, 2- ethyl-butyl, 2- methyl amyl, 3- methyl amyl, 4- methyl amyl, 2,3- dimethylbutyl, n-heptyl, 2- methyl oneself Base, 3- methylhexyl, 4- methylhexyl, 5- methylhexyl, 2,3- dimethyl amyl group, 2,4- dimethyl amyl group, 2,2- dimethyl Amyl, 3,3- dimethyl amyl group, 2- ethylpentyl, 3- ethylpentyl, n-octyl, 2,3- dimethylhexanyl, 2,4- dimethyl oneself Base, 2,5- dimethylhexanyl, 2,2- dimethylhexanyl, 3,3- dimethylhexanyl, 4,4- dimethylhexanyl, 2- ethylhexyl, 3- Ethylhexyl, 4- ethylhexyl, 2- methyl -2- ethylpentyl, 2- methyl -3- ethylpentyl, n-nonyl, 2- methyl -2- ethyl Hexyl, 2- methyl -3- ethylhexyl, 2,2- diethyl amyl group, positive decyl, 3,3- diethylhexyl, 2,2- diethylhexyl, and Its various branched isomer etc..Low alkyl group more preferably containing 1 to 6 carbon atom, non-limiting embodiment include first Base, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl, sec-butyl, n-pentyl, 1,1- dimethyl propyl, 1,2- Dimethyl propyl, 2,2- dimethyl propyl, 1- ethyl propyl, 2- methyl butyl, 3- methyl butyl, n-hexyl, 1- ethyl -2- first Base propyl, 1,1,2- thmethylpropyl, 1,1- dimethylbutyl, 1,2- dimethylbutyl, 2,2- dimethylbutyl, 1,3- diformazan Base butyl, 2- ethyl-butyl, 2- methyl amyl, 3- methyl amyl, 4- methyl amyl, 2,3- dimethylbutyl etc..Alkyl can be with It is substitution or non-substituted, when substituted, substituent group can be substituted on any workable tie point, the substitution Base is preferably one or more following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, Halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkanes Sulfenyl, heterocycle alkylthio group, oxo base ,-C (O) OR⁷、-OC(O)R⁷、-NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O) OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹Or-C (O) NR⁸R⁹。

Term " naphthenic base " refers to the unsaturated monocycle of saturation or part or polycyclic cyclic hydrocarbon substituent, cycloalkyl ring include 3 to 20 carbon atoms, preferably comprise 3 to 12 carbon atoms, more preferably include 3 to 10 carbon atoms, further preferably include 3~6 Carbon atom.The non-limiting example of monocyclic cycloalkyl includes cyclopropyl, cyclobutyl, cyclopenta, cyclopentenyl, cyclohexyl, hexamethylene Alkenyl, cyclohexadienyl, suberyl, cycloheptatriene base, cyclooctyl etc.；Polycyclic naphthene base includes the ring of loop coil, condensed ring and bridged ring Alkyl.

Term " spiro cycloalkyl group " refers to the polycyclic moiety that a carbon atom (claiming spiro-atom) is shared between 5 to 20 yuan of monocycle, It can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more Preferably 7 to 10 yuan.Spiro cycloalkyl group is divided into single spiro cycloalkyl group, double spirocyclanes according to the number for sharing spiro-atom between ring and ring Base or more spiro cycloalkyl groups, preferably single spiro cycloalkyl group and double spiro cycloalkyl groups.More preferably 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 Member/5 yuan or 5 yuan/6 yuan single spiro cycloalkyl groups.The non-limiting example of spiro cycloalkyl group includes:

Term " cycloalkyl " refers to 5 to 20 yuan, each ring in system and the shared a pair adjoined of other rings in system The full carbon polycyclic moiety of carbon atom, wherein one or more rings can be containing one or more double bonds, but none ring has The pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.It can be divided into according to a group cyclic number double Ring, tricyclic, Fourth Ring or polycyclic fused ring alkyl, preferably bicyclic or tricyclic, more preferably 5 yuan/5 yuan or 5 yuan/6 membered bicyclic alkyl. The non-limiting example of cycloalkyl includes:

Term " bridge ring alkyl " refers to 5 to 20 yuan, and the full carbon that any two ring shares two carbon atoms being not directly connected is more Cyclic group, can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic bridge ring alkyl can be divided into according to a group cyclic number, it is excellent It is selected as bicyclic, tricyclic or Fourth Ring, is more selected as bicyclic or tricyclic.The non-limiting example of bridge ring alkyl includes:

The cycloalkyl ring can be condensed on aryl, heteroaryl or heterocycloalkyl ring, wherein being connected to precursor structure Ring together is naphthenic base, and non-limiting example includes indanyl, tetralyl, benzocyclohepta alkyl etc..Naphthenic base can be Optionally replacing or non-substituted, when substituted, substituent group is preferably one or more following groups, independently selected from alkane Base, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, Aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, oxo base ,-C (O) OR⁷、-OC(O)R⁷、- NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O)OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹Or-C (O) NR⁸R⁹。

Term " heterocycle " refers to the unsaturated monocycle of saturation or part or polycyclic cyclic hydrocarbon substituent, and it includes 3 to 20 rings Atom, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O)_mThe hetero atom of (wherein m is integer 0 to 2), but do not wrap The loop section of-O-O- ,-O-S- or-S-S- are included, remaining annular atom is carbon.3 to 12 annular atoms are preferably comprised, wherein 1~4 It is hetero atom；It more preferably include 3 to 10 annular atoms；It more preferably include 3 to 8 annular atoms；It more preferably include 3 to 6 ring originals Son.The non-limiting example of monocyclic heterocycles base includes pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thio-morpholinyl, high piperazine Piperazine base etc..Multiring heterocyclic includes the heterocycle of loop coil, condensed ring and bridged ring.

Term " spiro heterocyclic radical " refers to the multiring heterocyclic that an atom (claiming spiro-atom) is shared between 5 to 20 yuan of monocycle Group, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O)_mThe hetero atom of (wherein m is integer 0 to 2), remaining annular atom For carbon.It can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 Member, more preferably 7 to 10 yuan.Spiro heterocyclic radical is divided into single spiro heterocyclic radical, double according to the number for sharing spiro-atom between ring and ring Spiro heterocyclic radical or more spiro heterocyclic radicals, preferably single spiro heterocyclic radical and double spiro heterocyclic radicals.More preferably 4 yuan/4 yuan, 4 yuan/5 yuan, 4 Member/6 yuan, 5 yuan/5 yuan or 5 yuan/6 yuan single spiro heterocyclic radicals.The non-limiting example of spiro heterocyclic radical includes:

Term " condensed hetero ring base " refers to 5 to 20 yuan, each ring in system and the shared a pair adjoined of other rings in system The polycyclic heterocyclic group of atom, one or more rings can be containing one or more double bonds, but none ring is with completely total The pi-electron system of yoke, wherein one or more annular atoms are the miscellaneous original selected from nitrogen, oxygen or S (O) m (wherein m is integer 0 to 2) Son, remaining annular atom are carbon.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.According to a group cyclic number can be divided into it is bicyclic, Tricyclic, Fourth Ring or polycyclic condensed hetero ring base, preferably bicyclic or tricyclic, more preferably 5 yuan/5 yuan or 5 yuan/6 membered bicyclic condensed hetero rings Base.The non-limiting example of condensed hetero ring base includes:

Term " bridge heterocycle " refers to 5 to 14 yuan, and any two ring shares the polycyclic heterocycle of two atoms being not directly connected Group, can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated, one of them or Multiple annular atoms are selected from nitrogen, oxygen or S (O)_mThe hetero atom of (wherein m is integer 0 to 2), remaining annular atom are carbon.Preferably 6 To 14 yuan, more preferably 7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic bridge heterocycle can be divided into according to a group cyclic number, Bicyclic or tricyclic is more selected as at preferably bicyclic, tricyclic or Fourth Ring.The non-limiting example of bridge heterocycle includes:

The heterocyclic ring can be condensed on aryl, heteroaryl or cycloalkyl ring, wherein being connected to one with precursor structure The ring risen is heterocycle, and non-limiting example includes:

Deng.

Heterocycle can be it is optionally replacing or non-substituted, when substituted, substituent group be preferably it is one or more with Lower group, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, Cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, oxo Base ,-C (O) OR⁷、-OC(O)R⁷、-NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O)OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹ Or-C (O) NR⁸R⁹。

Term " aryl " refers to that 6 to 14 yuan of full carbon monocycles of the pi-electron system with conjugation or fused polycycle are (namely shared The ring of adjacent carbon atoms pair) group, preferably 6 to 10 yuan, such as phenyl and naphthalene.The aryl rings can be condensed in heteroaryl On base, heterocycle or cycloalkyl ring, wherein being aryl rings, non-limiting example packet with the ring that precursor structure links together It includes:

Aryl can be substituted or non-substituted, and when substituted, substituent group is preferably one or more following groups, It is independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, ring Alkyl, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group ,-C (O) OR⁷、- OC(O)R⁷、-NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O)OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹Or-C (O) NR⁸R⁹。

Term " heteroaryl " refers to the heteroaromatic system comprising 1 to 4 hetero atom, 5 to 14 annular atoms, and wherein hetero atom selects From oxygen, sulphur and nitrogen.Heteroaryl is preferably 5 to 10 yuan, more preferably 5- or 6-membered, such as furyl, thienyl, pyridyl group, pyrrole Cough up base, N- alkyl pyrrole radicals, pyrimidine radicals, pyrazinyl, imidazole radicals, tetrazole radical etc..The heteroaryl ring can be condensed in aryl, miscellaneous In ring group or cycloalkyl ring, wherein the ring to link together with precursor structure is heteroaryl ring, non-limiting example includes:

Heteroaryl can be it is optionally replacing or non-substituted, when substituted, substituent group be preferably it is one or more with Lower group, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, Cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group ,-C (O)OR⁷、-OC(O)R⁷、-NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O)OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹Or-C (O) NR⁸R⁹。

Term " alkoxy " refers to-O- (alkyl) and-O- (non-substituted naphthenic base), and wherein alkyl is as defined above. The non-limiting example of alkoxy includes: methoxyl group, ethyoxyl, propoxyl group, butoxy, cyclopropyl oxygroup, cyclobutoxy group, penta oxygen of ring Base, cyclohexyloxy.Alkoxy can be optionally replacing or non-substituted, and when substituted, substituent group is preferably one or more A following group, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, Nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkane sulphur Base ,-C (O) OR⁷、-OC(O)R⁷、-NHS(O)_pR⁷、-C(O)R⁷、-NHC(O)R⁷、-NHC(O)OR⁷、-NR⁸R⁹、-OC(O)NR⁸R⁹ Or-C (O) NR⁸R⁹。

Term " halogenated alkyl " refers to the alkyl replaced by one or more halogens, and wherein alkyl is as defined above.

Term " hydroxyalkyl " refers to the alkyl being optionally substituted by a hydroxyl group, and wherein alkyl is as defined above.

Term " hydroxyl " refers to-OH group.

Term " halogen " refers to fluorine, chlorine, bromine or iodine.

Term " amino " refers to-NH₂。

Term " cyano " refers to-CN.

Term " nitro " refers to-NO₂。

Term " oxo base " refers to=O.

Term " carboxyl " refers to-C (O) OH.

Term " carboxylate " refers to-C (O) O (alkyl) or-C (O) O (naphthenic base), and wherein alkyl is as defined above.

Term " carboxylic acid halides " refers to the compound containing-C (O)-halogen group.

Term " ethyleneoxy " refers to CH₂=CH-O-.

Term " methylene " refers to-CH₂-。

Term " ethylidene " refers to-(CH₂)₂-。

Term " propylidene " refers to-(CH₂)₃-。

Term " butylidene " refers to-(CH₂)₄-。

Term " alkenyl " refers to-CH=CH-.

Term " alkynyl " refers to-C ≡ C-.

" X is selected from A, B or C ", " X is selected from A, B and C ", " X A, B or C ", " X A, B and C " etc. are different in the present invention uses Language expresses identical meaning, i.e. expression X can be any one or a few in A, B, C.

" optional " or " optionally " mean event or environment described later can with but need not occur, which includes The occasion that the event or environment occur or do not occur.For example, meaning that alkyl can be with " optionally by alkyl-substituted heterocyclic group " But necessarily exist, the explanation include heterocyclic group by alkyl-substituted situation and heterocyclic group not by alkyl-substituted situation.

" substituted " refers to one or more hydrogen atoms in group, preferably at most 5, more preferably 1~3 hydrogen atom Replaced independently of one another by the substituent group of respective number.Self-evident, substituent group is only in their possible chemical position, this Field technical staff, which can determine in the case where not paying excessive make great efforts and (pass through experiment or theoretical), may or impossible take Generation.It may be unstable when for example, amino or hydroxyl with free hydrogen are in conjunction with the carbon atom with unsaturated (such as olefinic) key Fixed.

" pharmaceutical composition " indicate containing one or more compounds described herein or its physiologically/pharmaceutical salt or The mixture and other components such as physiology/pharmaceutical carrier and excipient of pro-drug and other chemical constituents.Medicine The purpose of compositions is the administration promoted to organism, plays bioactivity in turn conducive to the absorption of active constituent.

" officinal salt " refers to the salt of the compounds of this invention, has safety when this kind of salt is used in the mammalian body and has Effect property, and there is due bioactivity.

The synthetic method of the compounds of this invention

In order to complete the purpose of the present invention, the present invention adopts the following technical scheme:

The present invention leads to formula (I) compound represented or the preparation method of its salt, comprising the following steps:

Scheme one

In acid condition, general formula (Ia) compound is oxidized to general formula (Ib) compound；General formula (Ib) compound is in alkali It is reacted under the conditions of property with general formula (Ic) compound, obtains general formula (Id) compound；General formula (Id) compound is in heating, alkaline condition Lower cyclization, the preferred N of alkali under this condition, N '-carbonyl dimidazoles obtain general formula (Ie) compound；General formula (Ie) after cyclization is changed It closes object and sloughs the protecting group on amino in acid condition, obtain general formula (If) compound or its salt；General formula (If) compound or The aqueous slkali of its salt reacts to obtain general formula (Ig) compound with the alcoholic solution of chlorosulphonyl isocyanate at low temperature, and the alcoholic solution is excellent Select t-butanol solution；General formula (Ig) compound sloughs the protecting group on amino in acid condition, obtains logical formula (IV) compound； Open loop obtains the logical formula (I) compound of target to obtained logical formula (IV) compound under alkaline condition.

The reagent for providing alkaline condition includes organic base and inorganic base, and the organic bases include but is not limited to pregnancy Two silicon substrate Sodamide of base, triethylamine, n,N-diisopropylethylamine, n-BuLi, potassium tert-butoxide, tetrabutylammonium bromide are described Inorganic base includes but is not limited to sodium hydride, sodium carbonate, sodium bicarbonate, potassium carbonate, saleratus or cesium carbonate.

Oxidant used includes but is not limited to: hydrogen peroxide, potassium permanganate and manganese dioxide.

Solvent for use includes but is not limited to: n,N-Dimethylformamide, toluene, acetic acid, methanol, ethyl alcohol, tetrahydrofuran, two Chloromethanes, dimethyl sulfoxide, Isosorbide-5-Nitrae-dioxane or water.

Wherein:

R¹~R⁴, m, n definition as described in logical formula (I).

The present invention leads to formula (II) compound represented or the preparation method of its salt, comprising the following steps:

Scheme two

In acid condition, general formula (Ia) compound is oxidized to general formula (Ib) compound；General formula (Ib) compound is in alkali It is reacted under the conditions of property with general formula (IIc) compound, obtains general formula (IId) compound；General formula (IId) compound is in heating, alkalinity Under the conditions of cyclization, the preferred N of alkali under this condition, N '-carbonyl dimidazoles obtain general formula (IIe) compound；General formula after cyclization (IIe) compound sloughs the protecting group on amino in acid condition, obtains general formula (IIf) compound or its salt；General formula (IIf) The aqueous slkali of compound or its salt reacts to obtain general formula (IIg) compound with the alcoholic solution of chlorosulphonyl isocyanate at low temperature, The preferred t-butanol solution of the alcoholic solution；General formula (IIg) compound sloughs the protecting group on amino in acid condition, obtains general formula (V) compound；Open loop obtains the logical formula (II) compound of target to obtained logical formula (V) compound under alkaline condition.

The reagent of alkaline condition includes organic base and inorganic base, and the organic bases include but is not limited to hexamethyl two Silicon substrate Sodamide, triethylamine, n,N-diisopropylethylamine, n-BuLi, potassium tert-butoxide, tetrabutylammonium bromide, described is inorganic Bases includes but is not limited to sodium hydride, sodium carbonate, sodium bicarbonate, potassium carbonate, saleratus or cesium carbonate.

Oxidant used includes but is not limited to: hydrogen peroxide, potassium permanganate and manganese dioxide.

Solvent for use includes but is not limited to: n,N-Dimethylformamide, toluene, acetic acid, methanol, ethyl alcohol, tetrahydrofuran, two Chloromethanes, dimethyl sulfoxide, Isosorbide-5-Nitrae-dioxane or water.

Wherein:

R¹、R³、R⁴, m, n definition as described in logical formula (I).

The present invention leads to formula (III) compound represented or the preparation method of its salt, comprising the following steps:

Scheme three

In acid condition, general formula (Ia) compound is oxidized to general formula (Ib) compound；General formula (Ib) compound is in alkali It is reacted under the conditions of property with general formula (IIIc) compound, obtains general formula (IIId) compound；General formula (IIId) compound is in heating, alkali Property under the conditions of cyclization, the preferred N of alkali under this condition, N '-carbonyl dimidazoles obtain general formula (IIIe) compound；It is logical after cyclization Formula (IIIe) compound sloughs the protecting group on amino in acid condition, obtains general formula (IIIf) compound or its salt；General formula (IIIf) aqueous slkali of compound or its salt reacts to obtain general formula (IIIg) with the alcoholic solution of chlorosulphonyl isocyanate at low temperature Compound, the preferred t-butanol solution of the alcoholic solution；General formula (IIIg) compound sloughs the protecting group on amino in acid condition, Obtain logical formula (VI) compound；Open loop obtains the logical formula (III) chemical combination of target to obtained logical formula (VI) compound under alkaline condition Object.

The reagent of alkaline condition includes organic base and inorganic base, and the organic bases include but is not limited to hexamethyl two Silicon substrate Sodamide, triethylamine, n,N-diisopropylethylamine, n-BuLi, potassium tert-butoxide, tetrabutylammonium bromide, described is inorganic Bases includes but is not limited to sodium hydride, sodium carbonate, sodium bicarbonate, potassium carbonate, saleratus or cesium carbonate.

Oxidant used includes but is not limited to: hydrogen peroxide, potassium permanganate and manganese dioxide.

Solvent for use includes but is not limited to: n,N-Dimethylformamide, toluene, acetic acid, methanol, ethyl alcohol, tetrahydrofuran, two Chloromethanes, dimethyl sulfoxide, Isosorbide-5-Nitrae-dioxane or water.

Wherein:

A is 3~8 member rings, wherein the ring is selected from carbocyclic ring or heterocycle；

R³、R⁴, m, n definition as described in logical formula (I).

Specific embodiment

The present invention is further described with reference to embodiments, but these embodiments not limit the scope of the present invention.

Embodiment

The structure of compound is by nuclear magnetic resonance (NMR) or mass spectrum (MS) come what is determined.The measurement of NMR is to use Bruker AVANCE-400 nuclear magnetic resonance spectrometer, measurement solvent are deuterated dimethyl sulfoxide (DMSO-d₆), deuterated chloroform (CDCl₃), deuterated methanol (CD₃OD), it is inside designated as tetramethylsilane (TMS), chemical shift is with 10^-6(ppm) it is provided as unit.

The measurement of MS is with FINNIGAN LCQAd (ESI) mass spectrograph (manufacturer: Thermo, model: Finnigan LCQ advantage MAX)。

The measurement of HPLC uses Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18 150 × 4.6mm chromatography Column) and Waters 2695-2996 high pressure liquid chromatograph (Gimini 150 × 4.6mm of C18 chromatographic column).

Kinases average inhibition and IC₅₀The measurement of value is with NovoStar microplate reader (German BMG company).

Tlc silica gel plate uses Yantai Huanghai Sea HSGF254 or Qingdao GF254 silica gel plate, and thin-layered chromatography (TLC) makes The specification that silica gel plate uses is 0.15mm~0.2mm, thin-layer chromatography isolate and purify product use specification be 0.4mm~ 0.5mm silica gel plate.

Column chromatography is generally carrier using 200~300 mesh silica gel of the Yantai Huanghai Sea.

Known starting material of the invention can be used or be synthesized according to methods known in the art, or can purchase certainly ABCR GmbH&Co.KG, Acros Organnics, Aldrich Chemical Company, splendid remote chemical science and technology (Accela ChemBio Inc), up to the companies such as auspicious chemicals.

In embodiment unless otherwise specified, reaction carries out under argon atmospher or nitrogen atmosphere.

Argon atmospher or nitrogen atmosphere refer to that reaction flask connects the argon gas or nitrogen balloon of an about 1L volume.

Nitrogen atmosphere refers to that reaction flask connects the hydrogen balloon of an about 1L volume.

Pressure hydration reaction uses Parr 3916EKX type hydrogenation instrument and clear indigo plant QL-500 type hydrogen generator or HC2-SS Type hydrogenates instrument.

Hydrogenation usually vacuumizes, and is filled with hydrogen, operates 3 times repeatedly.

Microwave reaction uses 908860 type microwave reactor of CEM Discover-S.

In embodiment unless otherwise specified, the solution in reaction refers to aqueous solution.

In embodiment unless otherwise specified, the temperature of reaction is room temperature.

Room temperature is optimum reaction temperature, and temperature range is 20 DEG C~30 DEG C.

The monitoring of reaction process in embodiment uses thin-layered chromatography (TLC), reacts the system of used solvent Have: A: methylene chloride and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, D: acetone, The volume ratio of solvent is different according to the polarity of compound and is adjusted.

The system of the solvent of the system and thin-layered chromatography of the eluant, eluent for the column chromatography that purifying compound uses includes: A: Methylene chloride and methanol system, B: n-hexane and ethyl acetate system, C: n-hexane, ethyl acetate and dichloromethane system, D: Petroleum ether and ethyl acetate system, E: the volume ratio of ethyl acetate, solvent is different according to the polarity of compound and is adjusted, A small amount of triethylamine and acid or alkaline reagent etc. can be added to be adjusted.

Embodiment 1

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- (((1- (sulphamoylamino) cyclopropyl) methyl) amino) -1,2, 5- oxadiazoles -3- carbonamidine 1

The first step

4- (the bromo- 4- fluorophenyl of 3-) -3- (4- nitro -1,2,5- oxadiazoles -3- base) -1,2,4- oxadiazoles -5 (4H) -one 1b

By 3- (4- amino -1,2,5- oxadiazoles -3- base) -4- (the bromo- 4- fluorophenyl of 3-) -1,2,4- oxadiazoles -5 (4H) - 5mL trifluoro is added in ketone 1a (264mg, 2.23mmol are prepared using method disclosed in patent application " WO2010005958 ") It in acetic acid, is added 3mL hydrogen peroxide (30%), is reacted 16 hours in 45 DEG C.After reaction, saturated sodium bisulfite solution is added to quench It goes out reaction, is extracted with ethyl acetate, merge organic phase, dried, filtered with anhydrous sodium sulfate, silicagel column is used in filtrate decompression concentration Chromatography purifies gained residue with eluant, eluent system B, obtains title product 1b (210mg, yellow oil), yield 78.6%.

MS m/z (LC-MS): 342.9 [M-1]

Second step

(1- (((4- (N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxy formamidine base) -1,2,5- oxadiazoles -3- bases) amino) first Base) cyclopropyl) t-butyl carbamate 1d

1b (210mg, 0.56mmol) is dissolved in 15mL tetrahydrofuran, (1- (aminomethyl) cyclopropyl) carbamic acid is added Tert-butyl ester 1c (210mg, 1.13mmol are prepared using method disclosed in patent application " WO2013052394 ") is added The sodium hydroxide solution of 1.5mL 2N reacts 30 minutes at room temperature.After reaction, the hydrochloric acid that 1N is added adjusts reaction solution pH To 2, it is extracted with ethyl acetate three times, merges organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration obtains crude product Title product 1d (260mg, yellow solid), product directly carry out next step reaction without further purification.

MS m/z (LC-MS): 485.2 [M+1]

Third step

(1- (((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- Oxadiazoles -3- base) amino) methyl) cyclopropyl) t-butyl carbamate 1e

Crude product 1d (260mg, 0.535mmol) is dissolved in 20mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (95mg, 0.586mmol), it is reacted 1 hour in 70 DEG C.After reaction, reaction solution is concentrated under reduced pressure, with ethyl acetate dissolution residual substance, Successively washed with 1N hydrochloric acid, water, saturated sodium chloride solution, with the dry organic phase of anhydrous sodium sulfate, filtering, filtrate decompression is concentrated, It obtains crude title product 1e (330mg, yellow solid), product directly carries out next step reaction without further purification.

MS m/z (LC-MS): 455.3 [M+1-56]

4th step

3- (4- (((1- amino cyclopropyl) methyl) amino) -1,2,5- oxadiazoles -3- bases) -4- (the bromo- 4- fluorophenyl of 3-) - 1,2,4- oxadiazoles -5 (4H) -one trifluoroacetate 1f

Crude product 1e (330mg, 0.645mmol) is dissolved in 30mL methylene chloride, 2mL trifluoroacetic acid is added, at room temperature Reaction 1 hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 1f (400mg, tan solid), Product directly carries out the next step without further purification.

5th step

N- (1- (((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2, 5- oxadiazoles -3- base) amino) methyl) cyclopropyl) sulphamoylamino t-butyl formate 1g

By chlorosulphonyl isocyanate (0.793g, 5.6mmol, using well known method " Organic Syntheses, 1966,46,23-7 " is prepared) it is dissolved in 10mL methylene chloride, it is cooled to 0 DEG C, is added the tert-butyl alcohol (0.43g, 5.8mmol), Reaction solution reacts 1 hour at 0 DEG C, and reaction solution A is made.Crude product 1f (400mg, 1.0mmol) is dissolved in 20mL methylene chloride, 0.75mL triethylamine is added, reaction solution B is made.At 0 DEG C, reaction solution A is added in reaction solution B, is reacted 1 hour in 0 DEG C.Instead After answering, saturated sodium bicarbonate solution is added, reaction solution is quenched, liquid separation, organic phase washed with water washing, saturated sodium-chloride Solution washing, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, with thin-layered chromatography with solvent system A purify obtained by it is residual Excess obtains title product 1g (120mg, white solid), yield 20%.

MS m/z (LC-MS): 607.1 [M+18]

6th step

N- 1- [(4- [4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base] -1,2, 5- oxadiazoles -3- base } amino) methyl] cyclopropyl } sulfamide 1h

1g (120mg, 0.198mmol) is dissolved in 20mL methylene chloride, 4mL trifluoroacetic acid is added, reacts 1 at room temperature Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 1h (160mg, brown viscous object), product is not Purified direct carry out the next step.

MS m/z (LC-MS): 490.0 [M+1]

7th step

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- (((1- (sulphamoylamino) cyclopropyl) methyl) amino) -1,2, 5- oxadiazoles -3- carbonamidine 1

Crude product 1h (160mg, 0.27mmol) is dissolved in 20mL methanol, is added potassium carbonate (230mg, 1.67mmol), in 50 DEG C are reacted 1 hour.After reaction, saturated ammonium chloride solution neutralization reaction, liquid separation is added, water phase is extracted with ethyl acetate three It is secondary, merge organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration is pure with solvent system A with thin-layered chromatography Change gained residue, obtains title product 1 (10mg, white solid), yield 7.5%.

MS m/z (ESI): 466.2 [M+1]

¹H NMR (400MHz, DMSO-d₆) δ 11.54 (s, 1H), 8.89 (s, 1H), 7.11-7.25 (m, 3H), 6.78- 6.79 (m, 1H), 6.68 (s, 2H), 6.23 (t, 1H), 3.44 (d, 2H), 0.94 (s, 2H), 0.68 (s, 2H)

Embodiment 2

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- (((1- ((sulphamoylamino) methyl) cyclopropyl) methyl) ammonia Base) -1,2,5- oxadiazoles -3- carbonamidines 2

The first step

((1- (((4- (N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxy formamidine base -1,2,5- oxadiazoles -3- base) amino) first Base) cyclopropyl) methyl) t-butyl carbamate 2b

1b (360mg, 0.968mmol) is dissolved in 30mL tetrahydrofuran, ((1- (amino methyl) cyclopropyl) first is added Base) (387mg, 1.935mmol are prepared t-butyl carbamate 2a using method disclosed in patent application " WO2009054468 " And obtain), the sodium hydroxide solution of 2.9mL 2N is added, reacts 30 minutes at room temperature.After reaction, the hydrochloric acid tune of 1N is added Reaction solution pH < 7 is saved, is extracted with ethyl acetate three times, is merged organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression is dense Contracting, obtains crude title product 2b (650mg, Tan solid), and product directly carries out next step reaction without further purification.

Second step

((1- (((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- Oxadiazoles -3- base) amino) methyl) cyclopropyl) methyl) t-butyl carbamate 2c

Crude product 2b (650mg, 1.3mmol) is dissolved in 30mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (222mg, 1.37mmol), it is reacted 1 hour in 70 DEG C.After reaction, reaction solution is concentrated under reduced pressure, with ethyl acetate dissolution residual substance, according to Secondary to be washed with 1N hydrochloric acid, water, saturated sodium chloride solution, with the dry organic phase of anhydrous sodium sulfate, filtering, filtrate decompression concentration is obtained To crude title product 2c (715mg, bright-yellow solid), product directly carries out next step reaction without further purification.

Third step

3- (4- (((1- (amino methyl) cyclopropyl) methyl) amino) -1,2,5- oxadiazoles -3- bases) -4- (bromo- 4- fluorine of 3- Phenyl) -5 (4H) -one 2d of -1,2,4- oxadiazoles

Crude product 2c (715mg, 1.36mmol) is dissolved in 5mL methylene chloride, 5mL trifluoroacetic acid is added, at room temperature instead It answers 1 hour.After reaction, reaction solution is concentrated under reduced pressure, is obtained crude title product 2d (1.02g, brown-red oil), produced Product directly carry out next step reaction without further purification.

MS m/z (LC-MS): 427.0 [M+1]

4th step

N- ((1- (((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2, 5- oxadiazoles -3- base) amino) methyl) cyclopropyl) methyl) sulphamoylamino t-butyl formate 2e

Chlorosulphonyl isocyanate (509mg, 3.59mmol) is dissolved in 10mL methylene chloride, is cooled to 0 DEG C, tertiary fourth is added Alcohol (267mg, 3.59mmol), reaction solution react 15 minutes at 0 DEG C, and reaction solution A is made.By crude product 2d (1.021g, It 2.4mmol) is dissolved in 20mL methylene chloride, 1mL triethylamine is added, reaction solution B is made.Reaction solution is added in reaction solution A at 0 DEG C In B, reacted 1 hour in 0 DEG C.After reaction, be added saturated sodium bicarbonate solution reaction solution is quenched, liquid separation, organic phase according to Secondary to be washed with water, saturated sodium chloride solution washing, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, with thin-layered chromatography with Solvent system A purifying gained residue, obtains title product 2e (28mg, brown solid), yield 19%.

MS m/z (LC-MS): 621.0 [M+18]

5th step

N- (1- [(4- [4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base] -1,2, 5- oxadiazoles -3- base } amino) methyl] cyclopropyl } methyl) sulfamide 2f

2e (282mg, 0.466mmol) is dissolved in 10mL methylene chloride, 5mL trifluoroacetic acid is added, reacts 1 at room temperature Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 2f (160mg, brown viscous object), product is not Purified direct carry out the next step.

MS m/z (LC-MS): 504.0 [M+1]

6th step

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- (((1- ((sulphamoylamino) methyl) cyclopropyl) methyl) ammonia Base) -1,2,5- oxadiazoles -3- carbonamidines 2

Crude product 2f (386mg, 0.765mmol) is dissolved in 10mL methanol, is added potassium carbonate (528mg, 3.86mmol), in 50 DEG C are reacted 1 hour.After reaction, saturated ammonium chloride solution neutralization reaction, liquid separation is added, water phase is extracted with ethyl acetate three It is secondary, merge organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration is pure with solvent system A with thin-layered chromatography Change gained residue, obtains title product 2 (180mg, white solid), yield 49%.

MS m/z (ESI): 478.2 [M+1]

¹H NMR (400MHz, DMSO-d₆) δ 11.47 (s, 1H), 8.89 (s, 1H), 7.19 (t, 1H), 7.10-7.12 (m, 1H), 6.78-6.79 (m, 1H), 6.77 (t, 1H), 6.49 (s, 2H), 6.14 (t, 1H), 3.23 (d, 2H), 2.86 (d, 2H), 0.48 (s, 4H)

Embodiment 3

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1, 2,5- oxadiazoles -3- carbonamidines 3

The first step

(1- (2- ((4- (N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxy formamidine base) -1,2,5- oxadiazoles -3- bases) amino) second Base) cyclopropyl) t-butyl carbamate 3b

1b (210mg, 0.564mmol) is dissolved in 20mL tetrahydrofuran, the tert-butyl ester (1- (2- amino-ethyl) cyclopropyl is added Base) carbamic acid 3a (240mg, 1.19mmol are prepared using method disclosed in patent application " WO2013020993 "), add Enter the sodium hydroxide solution of 1.5mL 2N, reacts 30 minutes at room temperature.After reaction, the hydrochloric acid that 1N is added adjusts reaction solution PH < 7 is extracted with ethyl acetate three times, merges organic phase, dried, filtered with anhydrous sodium sulfate, and filtrate decompression concentration obtains thick Product title product 3b (370mg, faint yellow solid), product directly carry out next step reaction without further purification.

MS m/z (LC-MS): 497.1 [M-1]

Second step

(1- (2- ((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2, 5- oxadiazoles -3- base) amino) ethyl) cyclopropyl) t-butyl carbamate 3c

Crude product 3b (370mg, 0.74mmol) is dissolved in 15mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (126mg, 0.78mmol), it is reacted 1 hour in 70 DEG C.After reaction, reaction solution is concentrated under reduced pressure, with ethyl acetate dissolution residual substance, according to Secondary to be washed with 1N hydrochloric acid, water, saturated sodium chloride solution, with the dry organic phase of anhydrous sodium sulfate, filtering, filtrate decompression concentration is obtained To crude title product 3c (350mg, yellow solid), product directly carries out next step reaction without further purification.

MS m/z (LC-MS): 523.0 [M-1]

Third step

3- (4- ((2- (1- amino cyclopropyl) ethyl) amino) -1,2,5- oxadiazoles -3- bases) -4- (bromo- 4- fluorobenzene of 3- Base) -5 (4H) -one 3d of -1,2,4- oxadiazoles

Crude product 3c (350mg, 0.666mmol) is dissolved in 5mL methylene chloride, 3mL trifluoroacetic acid is added, at room temperature instead It answers 1 hour.After reaction, reaction solution is concentrated under reduced pressure, is obtained crude title product 3d (300mg, tan solid), produced Product directly carry out the next step without further purification.

4th step

N- (1- (2- ((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1, 2,5- oxadiazoles -3- bases) amino) ethyl) cyclopropyl) sulphamoylamino t-butyl formate 3e

Chlorosulphonyl isocyanate (145mg, 1.024mmol) is dissolved in 10mL methylene chloride, is cooled to 0 DEG C, uncle is added Butanol (76mg, 1.025mmol), reaction solution react 15 minutes at 0 DEG C, and reaction solution A is made.By crude product 3d (500mg, It 1.17mmol) is dissolved in 20mL methylene chloride, 1mL triethylamine is added, reaction solution B is made.At 0 DEG C, reaction solution A is added and is reacted In liquid B, reacted 1 hour in 0 DEG C.After reaction, saturated sodium bicarbonate solution is added reaction solution is quenched, liquid separation, organic phase It is successively washed with water, saturated sodium chloride solution washing, anhydrous sodium sulfate dries, filters, and thin-layered chromatography is used in filtrate decompression concentration Gained residue is purified with solvent system A, obtains title product 3e (180mg, brown solid), yield 25%.

MS m/z (LC-MS): 602.0 [M-1]

5th step

N- 1- [2- (4- [4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base] -1, 2,5- oxadiazoles -3- bases } amino) ethyl] cyclopropyl } sulfamide 3f

3e (180mg, 0.298mmol) is dissolved in 40mL methylene chloride, 2mL trifluoroacetic acid is added, reacts 1 at room temperature Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 3f (223mg, isabelline dope), product The next step is directly carried out without further purification.

6th step

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1, 2,5- oxadiazoles -3- carbonamidines 3

Crude product 3f (223mg, 0.44mmol) is dissolved in 15mL methanol, is added potassium carbonate (380mg, 2.75mmol), in 50 DEG C are reacted 1 hour.After reaction, saturated ammonium chloride solution neutralization reaction, liquid separation is added, water phase is extracted with ethyl acetate three It is secondary, merge organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration is pure with solvent system A with thin-layered chromatography Change gained residue, obtains title product 3 (70mg, white solid), yield 33%.

MS m/z (ESI): 480.1 [M+1]

¹H NMR (400MHz, DMSO-d₆) δ 11.45 (s, 1H), 8.89 (s, 1H), 7.19 (t, 1H), 7.07-7.10 (m, 2H), 6.77-6.78 (m, 1H), 6.60 (s, 2H), 6.13 (t, 1H), 3.38-3.43 (m, 2H), 1.82 (t, 2H), 0.88 (t, 2H), 0.46 (t, 2H)

Embodiment 4

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((3- hydroxyl -2- (sulphamoylamino) propyl) amino) -1,2, 5- oxadiazoles -3- carbonamidine 4

The first step

4- (((4- (N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxy formamidine base) -1,2,5- oxadiazoles -3- bases) amino) first Base) -2,2- dimethyl oxazolidine -3- carboxylic acid tert-butyl ester 4b

1b (505mg, 1.54mmol) is dissolved in 30mL tetrahydrofuran, 4- (amino methyl) -2,2- dimethyl is added and dislikes Oxazolidine -3- carboxylic acid tert-butyl ester 4a (630mg, 2.73mmol, using the preparation of method disclosed in patent application " WO2013020993 " ), the sodium hydroxide solution of 4mL 2N is added, reacts 30 minutes at room temperature.After reaction, saturated ammonium chloride solution is added Adjusting pH value of solution is 9, is extracted with ethyl acetate three times, merges organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression is dense Contracting, obtains crude title product 4b (667mg, brown solid), and product directly carries out next step reaction without further purification.

MS m/z (LC-MS): 529.1 [M-1]

Second step

(((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 4- Diazole -3- base) amino) methyl) -2,2- dimethyl oxazolidine -3- carboxylic acid tert-butyl ester 4c

Crude product 4b (667mg, 1.26mmol) is dissolved in 20mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (225mg, 1.38mmol), it is reacted 1 hour in 70 DEG C.After reaction, reaction solution is concentrated under reduced pressure, with ethyl acetate dissolution residual substance, according to Secondary to be washed with 1N hydrochloric acid, water, saturated sodium chloride solution, with the dry organic phase of anhydrous sodium sulfate, filtering, filtrate decompression concentration is obtained To crude title product 4c (710mg, tan solid), product directly carries out next step reaction without further purification.

MS m/z (LC-MS): 555.1 [M-1]

Third step

(((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 1- Diazole -3- base) amino) -3- hydroxy propane -2- base) t-butyl carbamate 4d

Crude product 4c (710mg, 1.28mmol) is dissolved in 20mL methanol, addition p-methyl benzenesulfonic acid monohydrate (124mg, 0.65mmol), it reacts 12 hours at room temperature.After reaction, saturated sodium bicarbonate solution neutralization reaction is added, by reaction solution It is concentrated under reduced pressure, is extracted with ethyl acetate three times, merges organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration is used Thin-layered chromatography purifies gained residue with solvent system A, obtains title product 4d (310mg, tan solid), yield 47%.

MS m/z (LC-MS): 513.0 [M-1]

4th step

((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 3- Diazole -3- base) amino) -2- ((tertbutyloxycarbonyl) amino) propyl-acetic acid ester 4e

4d (310mg, 0.60mmol) is dissolved in 30mL methylene chloride, 0.18mL triethylamine is added, is cooled to 0 DEG C, drop Add chloroacetic chloride (0.066mL, 0.92mmol), is reacted 1 hour in 0 DEG C.After reaction, saturated sodium bicarbonate solution is added, point Liquid, organic phase washed with water, saturated sodium chloride solution washing, anhydrous sodium sulfate dry, filter, and filtrate decompression concentration obtains thick Product title product 4e (510mg, brown viscous object), product directly carry out the next step without further purification.

MS m/z (LC-MS): 556.0 [M-1]

5th step

2- amino -3- ((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1, 2,5- oxadiazoles -3- bases) amino) propyl-acetic acid ester trifluoroacetate 4f

Crude product 4e (300mg, 0.538mmol) is dissolved in 5mL methylene chloride, 3mL trifluoroacetic acid is added, in room temperature reaction 1 Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 4f (510mg, tan solid), product It directly carries out without further purification in next step.

6th step

((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 3- Diazole -3- base) amino) -2- ((N- (tertbutyloxycarbonyl) sulfamoyl) amino) propyl-acetic acid ester 4g

Chlorosulphonyl isocyanate (120mg, 0.848mmol) is dissolved in 10mL methylene chloride, is cooled to 0 DEG C, uncle is added Butanol (63mg, 0.849mmol), reaction solution react 15 minutes in 0 DEG C, and reaction solution A is made.By crude product 4f (510mg, It 1.11mmol) is dissolved in 10mL methylene chloride, 1mL triethylamine is added, reaction solution B is made.Reaction solution A is added in room temperature and is reacted In liquid B, in room temperature reaction 1 hour.After reaction, saturated sodium bicarbonate solution is added reaction solution is quenched, liquid separation organic phase It is successively washed with water, saturated sodium chloride solution, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, with thin-layered chromatography to open up Agent system A purifying gained residue is opened, title product 4g (238mg, yellow solid), yield 33% are obtained.

MS m/z (LC-MS): 653.0 [M+18]

7th step

((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 3- Diazole -3- base) amino) -2- (sulphamoylamino) propyl-acetic acid ester 4h

4g (238mg, 0.37mmol) is dissolved in 5mL methylene chloride, 3mL trifluoroacetic acid is added, it is small to react 1 at room temperature When.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 4h (250mg, filbert dope), product is not Purified direct carry out the next step.

8th step

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((3- hydroxyl -2- (sulphamoylamino) propyl) amino) -1,2, 5- oxadiazoles -3- carbonamidine 4

Crude product 4h (250mg, 0.666mmol) is dissolved in 20mL methanol, is added potassium carbonate (600mg, 4.35mmol), in 50 DEG C are reacted 1.5 hours.After reaction, reaction solution is concentrated under reduced pressure, with thin-layered chromatography with solvent system A purify gained Residue obtains title product 4 (40mg, faint yellow solid), yield 18%.

MS m/z (ESI): 468.1 [M+1]

¹H NMR (400MHz, DMSO-d₆) δ 11.47 (s, 1H), 8.87 (s, 1H), 7.12-7.19 (m, 2H), 6.77- 6.78 (m, 2H), 6.61 (d, 1H), 6.56 (s, 1H), 6.20-6.22 (m, 1H), 4.82 (t, 1H), 4.02-4.04 (m, 1H), 3.44-3.57 (m, 3H), 3.24-3.26 (m, 1H)

Embodiment 5

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((3- methoxyl group -2- (sulphamoylamino) propyl) amino) -1, 2,5- oxadiazoles -3- carbonamidines 5

The first step

(1- ((4- (N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxy formamidine base) -1,2,5- oxadiazoles -3- bases) amino) -3- first Oxygroup propane -2- base) t-butyl carbamate 5b

1b (756mg, 2.03mmol) is dissolved in 50mL tetrahydrofuran, is added (1- amino -3- methoxy propane -2- base) T-butyl carbamate 5a (800mg, 3.92mmol, using the preparation of method disclosed in patent application " WO2012002577 " ), the sodium hydroxide solution of 6mL 2N is added, in room temperature reaction 30 minutes.After reaction, the hydrochloric acid that 1N is added adjusts reaction Liquid is extracted with ethyl acetate three times to pH < 7, merges organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration obtains To crude title product 5b (1g, yellow solid), product directly carries out next step reaction without further purification.

MS m/z (LC-MS): 475.1 [M+1-28]

Second step

(((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 1- Diazole -3- base) amino) -3- methoxy propane -2- alkane) t-butyl carbamate 5c

Crude product 5b (1g, 1.99mmol) is dissolved in 30mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (322mg, 1.99mmol), it is reacted 1 hour in 70 DEG C.After reaction, reaction solution is concentrated under reduced pressure, with ethyl acetate dissolution residual substance, according to Secondary to be washed with 1N hydrochloric acid, water, saturated sodium chloride solution, with the dry organic phase of anhydrous sodium sulfate, filtering, filtrate decompression concentration is obtained To crude title product 5c (1g, yellow solid), product directly carries out next step reaction without further purification.

Third step

3- (4- ((2- amino -3- methoxy-propyl) amino) -1,2,5- oxadiazoles -3- bases) -4- (bromo- 4- fluorobenzene of 3- Base) -5 (4H) -one 5d of -1,2,4- oxadiazoles

Crude product 5c (1g, 1.89mmol) is dissolved in 10mL methylene chloride, 10mL trifluoroacetic acid is added, in room temperature reaction 1 Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 5d (1.55g, tan solid), product The next step is directly carried out without further purification.

MS m/z (LC-MS): 429.1 [M+1]

4th step

N- (1- ((4- (4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- Oxadiazoles -3- base) amino) -3- methoxy propane -2- base) sulphamoylamino t-butyl formate 5e

Chlorosulphonyl isocyanate (400mg, 2.83mmol) is dissolved in 30mL methylene chloride, is cooled to 0 DEG C, tertiary fourth is added Alcohol (210mg, 2.83mmol), reaction solution is in 0 DEG C of reaction, 15 minutes obtained reaction solution A.Crude product 5d (1.55g, 3.6mmol) is molten In 20mL methylene chloride, it is cooled to 0 DEG C, 2.5mL triethylamine is added, reaction solution B is made.Reaction solution A is added dropwise to instead in room temperature It answers in liquid B, in room temperature reaction 1 hour.After reaction, saturated sodium bicarbonate solution is added reaction solution is quenched, liquid separation is organic Mutually successively washed with water, saturated sodium chloride solution, anhydrous sodium sulfate dries, filters, filtrate decompression be concentrated, with thin-layered chromatography with Solvent system A purifying gained residue, obtains title product 5e (1.3g, Tan solid), yield 59%.

MS m/z (LC-MS): 625.0 [M+18]

5th step

N- [1- ({ 4- [4- (the bromo- 4- fluorophenyl of 3-) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base] -1,2,5- Oxadiazoles -3- base } amino) -3- methoxy propane -2- base] sulfamide 5f

5e (410mg, 0.67mmol) is dissolved in 10mL methylene chloride, 8mL trifluoroacetic acid is added, reacts 1 at room temperature Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 5f (574mg, brown oil), product is not Purified direct carry out the next step.

6th step

N- (the bromo- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((3- methoxyl group -2- (sulphamoylamino) propyl) amino) -1, 2,5- oxadiazoles -3- carbonamidines 5

Crude product 5f (574mg, 1.129mmol) is dissolved in 10mL tetrahydrofuran, the sodium hydroxide solution of 3mL 2N is added, In room temperature reaction 15 minutes.After reaction, saturated ammonium chloride solution neutralization reaction, liquid separation is added, water phase is extracted with ethyl acetate It takes three times, merges organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration, with thin-layered chromatography with solvent system A purifying gained residue, obtains title product 5 (150mg, white solid), yield 27%.

MS m/z (ESI): 484.1 [M+1]

¹H NMR (400MHz, DMSO-d₆) δ 11.45 (s, 1H), 8.88 (s, 1H), 7.12-7.19 (m, 2H), 6.74- 6.78 (m, 2H), 6.59 (s, 2H), 6.20-6.2 (m, 1H), 3.61-3.63 (m, 1H), 3.43-3.46 (m, 2H), 3.25- 3.36 (m, 2H), 3.28 (s, 3H)

Embodiment 6

N- (3- chlorphenyl)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1,2,5- Oxadiazoles -3- carbonamidine 6

The first step

4- amino-N- (3- chlorphenyl)-N '-hydroxyl -1,2,5- oxadiazoles -3- carbonamidine 6b

3- chloroaniline (4.7g, 36.8mmol) is dissolved in 100mL ethyl alcohol, prefabricated 50mL 4- Amino-N-hydroxy 1 is added, 2,5- oxadiazoles -3- carbimide base chlorine 6a (5g, 30.8mmol, using well known method " Journal of the American Chemical Society, 2015,137 (33), 10532-10535 " are prepared) ethyl alcohol suspension, be added dropwise Triethylamine (5.1mL, 36.7mmol) is stirred to react 12 hours.After reaction, reaction solution is concentrated under reduced pressure, with silicagel column color Spectrometry purifies gained residue with eluant, eluent system B, obtains title product 6b (6.0g, faint yellow solid), yield 76.9%.

Second step

- 5 (4H) -one 6c of 3- (4- amino -1,2,5- oxadiazoles -3- base) -4- (3- chlorphenyl) -1,2,4- oxadiazoles

6b (5g, 19mmol) is dissolved in 100mL tetrahydrofuran, addition N, N '-carbonyl dimidazoles (3.2g, 19mmol), It is reacted 1 hour at 70 DEG C.After reaction, reaction solution is cooled to room temperature, is concentrated under reduced pressure, ethyl acetate and 1N salt is added Acid, liquid separation wash organic phase with saturated sodium chloride solution, and organic phase dries, filters removing desiccant, filtrate with anhydrous sodium sulfate It is concentrated under reduced pressure, is beaten to obtain crude title product 6c (3.7g, khaki solid) with ethyl acetate and n-hexane, product is without pure Change and directly carries out the next step.

Third step

- 5 (4H) -one 6d of 4- (3- chlorphenyl) -3- (4- nitro -1,2,5- oxadiazoles -3- base) -1,2,4- oxadiazoles

Crude product 6c (800mg, 2.86mmol) is dissolved in 15mL trifluoroacetic acid, is added 7mL hydrogen peroxide (30%), in 50 DEG C Reaction 12 hours.After reaction, ethyl acetate is added, adds saturated sodium bisulfite solution, liquid separation, water phase ethyl acetate Extraction three times, merge organic phase, dry, filter removing desiccant with anhydrous sodium sulfate, be concentrated under reduced pressure, with silica gel column chromatography with Eluant, eluent system B purifying gained residue, obtains title product 6d (320mg, faint yellow solid), yield 36%.

4th step

((((4- (4- (3- chlorphenyl) -5- carbonyl -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- dislikes two to 2- to 1- Azoles -3- base) amino) ethyl) cyclopropyl) t-butyl carbamate 6e

6d (320mg, 1.03mmol) and 3a (227mg, 1.135mmol) are dissolved in 20mL tetrahydrofuran, 1.1mL is added The sodium hydroxide solution of 1N is stirred to react 30 minutes.After reaction, the hydrochloric acid that 1N is added adjusts reaction solution pH < 7, and second is added Acetoacetic ester, liquid separation, water phase are extracted with ethyl acetate three times, merge organic phase, dried, filtered with anhydrous sodium sulfate, filtrate decompression Concentration, obtains crude title product 6e (540mg, faint yellow solid), and product directly carries out next step reaction without further purification.

5th step

3- (4- ((2- (1- amino cyclopropyl) ethyl) amino) -1,2,5- oxadiazoles -3- bases) -4- (3- chlorphenyl) -1, - 5 (4H) -one 6f of 2,4- oxadiazoles

Crude product 6e (540mg, 1.17mmol) is dissolved in 15mL methylene chloride, 10mL trifluoroacetic acid is added and is stirred to react 1 Hour.After reaction, reaction solution is concentrated under reduced pressure, obtains crude title product 6f (900mg, brown oil), product is not It is purified directly to carry out next step reaction.

6th step

((((4- (4- (3- chlorphenyl) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base) -1,2,5- is disliked 2- 1- N- Diazole -3- base) amino) ethyl) cyclopropyl) sulphamoylamino t-butyl formate 6g

Chlorosulphonyl isocyanate (215mg, 1.52mmol) is dissolved in 15mL methylene chloride, is cooled to 0 DEG C, tertiary fourth is added Alcohol (113mg, 1.52mmol), reaction solution react 15 minutes at 0 DEG C, and reaction solution A is made.By crude product 6f (900mg, It 2.48mmol) is dissolved in 30mL methylene chloride, 1mL triethylamine is added, reaction solution B is made.Reaction solution A is added at 0 DEG C and is reacted In liquid B, react 1 hour.After reaction, saturated sodium bicarbonate solution is added reaction is quenched, liquid separation, organic phase washed with water Washing, saturated sodium chloride solution washing, anhydrous sodium sulfate dry, filter, filtrate decompression concentration, with silica gel column chromatography to elute Agent system B purifying gained residue, obtains title product 6g (270mg, white solid), yield 20%.

7th step

{ [({ 4- [4- (3- chlorphenyl) -5- oxo -4,5- dihydro -1,2,4- oxadiazoles -3- base] -1,2,5- is disliked 2- 1- N- Diazole -3- base } amino) ethyl] cyclopropyl } sulfamide 6h

6g (270mg, 0.498mmol) is dissolved in 10mL methylene chloride, 5mL trifluoroacetic acid is added, it is small to be stirred to react 1 When.After reaction, reaction solution is concentrated under reduced pressure, is obtained crude title product 6h (264mg, faint yellow solid), product without Purifying directly carries out the next step.

8th step

N- (3- chlorphenyl)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1,2,5- Oxadiazoles -3- carbonamidine 6

Crude product 6h (264mg, 0.597mmol) is dissolved in 30mL tetrahydrofuran, the sodium hydroxide of 1.2mL 2.5M is added Solution is stirred to react 15 minutes.After reaction, saturated ammonium chloride solution, liquid separation is added, water phase is extracted with ethyl acetate three It is secondary, merge organic phase, is dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration, with silica gel column chromatography with eluant, eluent system B Purifying gained residue, obtains title product 6 (50mg, white solid), yield 20%.

MS m/z (ESI): 414.3 [M+1]

¹H NMR (400MHz, CD₃OD) δ 7.17 (t, 1H), 6.97-6.99 (m, 1H), 6.86 (t, 1H), 6.74-6.76 (m, 1H), 3.56 (t, 2H), 1.93 (t, 2H), 1.00-1.03 (m, 2H), 0.59-0.62 (m, 2H)

Embodiment 7

N- (4- fluoro- 3- (trifluoromethyl) phenyl)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) Amino) -1,2,5- oxadiazoles -3- carbonamidines 7

Using the synthetic route of embodiment 6, first step raw material is replaced with into 6a and 4- fluoro- 3- (trifluoromethyl) aniline, is made It obtains title product 7 (170mg, white solid).

MS m/z (ESI): 468.3 [M+1]

¹H NMR (400MHz, CD₃OD) δ 7.14-7.20 (m, 2H), 7.10-7.12 (m, 1H), 3.56 (t, 2H), 1.94 (t, 2H), 1.00-1.03 (m, 2H), 0.60-0.63 (m, 2H)

Embodiment 8

N- (the fluoro- 3- aminomethyl phenyl of 4-)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) - 1,2,5- oxadiazoles -3- carbonamidine 8

Using the synthetic route of embodiment 6, first step raw material is replaced with into the fluoro- 3- methylaniline of 6a and 4-, title is made Product 8 (150mg, white solid).

MS m/z (ESI): 414.3 [M+1]

¹H NMR (400MHz, CD₃OD) δ 6.87 (t, 1H), 6.78-6.81 (m, 1H), 6.68-6.72 (m, 1H), 3.53 (d, 2H), 2.20 (d, 3H), 1.91 (t, 2H), 0.99-1.02 (m, 2H), 0.58-0.61 (m, 2H)

Embodiment 9

N- (3- bromophenyl)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1,2,5- Oxadiazoles -3- carbonamidine 9

Using the synthetic route of embodiment 6, first step raw material is replaced with into 6a and 3- bromaniline, title product 9 is made (50mg, white solid).

MS m/z (ESI): 462.2 [M+1]

¹H NMR (400MHz, CD₃OD) δ 7.11-7.12 (m, 2H), 7.01-7.02 (m, 1H), 6.78-6.80 (m, 1H), 3.55 (t, 2H), 1.93 (t, 2H), 1.00-1.03 (m, 2H), 0.59-0.62 (m, 2H)

Embodiment 10

N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino)-N- (m- tolyl) -1,2,5- Oxadiazoles -3- carbonamidine 10

Using the synthetic route of embodiment 6, first step raw material is replaced with into 6a and meta-aminotoluene, title product 10 is made (250mg, white solid).

MS m/z (ESI): 396.3 [M+1]

¹H NMR (400MHz, CD₃OD) δ 7.08 (t, 1H), 6.84 (d, 1H), 6.64 (s, 1H), 6.63 (d, 1H), 3.52 (t, 2H), 2.27 (s, 3H), 1.90 (t, 2H), 0.98-1.01 (m, 2H), 0.57-0.60 (m, 2H)

Embodiment 11

N- (the chloro- 4- fluorophenyl of 3-)-N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino) -1, 2,5- oxadiazoles -3- carbonamidines 11

Using the synthetic route of embodiment 6, first step raw material is replaced with into the chloro- 4- fluoroaniline of 6a and 3-, title is made and produces Object 11 (30mg, brown solid).

MS m/z (ESI): 434.2 [M+1]

¹H NMR (400MHz, CD₃0D) δ 7.08 (t, 1H), 6.98-6.99 (m, 1H), 6.80-6.82 (m, 1H), 3.55 (t, 2H), 1.93 (t, 2H), 1.01 (s, 2H), 0.61 (s, 2H)

Embodiment 12

N '-hydroxyl -4- ((2- (1- (sulphamoylamino) cyclopropyl) ethyl) amino)-N- (3- (trifluoromethyl) benzene Base) -1,2,5- oxadiazoles -3- carbonamidines 12

Using the synthetic route of embodiment 6, first step raw material is replaced with into 6a and 3- (trifluoromethyl) aniline, title is made Product 12 (105mg, white solid).

MS m/z (ESI): 450.3 [M+1]

¹H NMR (400MHz, CD₃OD) δ 7.38 (t, 1H), 7.25 (d, 1H), 7.10 (s, 1H), 7.05 (d, 1H), 3.57 (t, 2H), 1.94 (t, 2H), 1.00-1.03 (m, 2H), 0.59-0.62 (m, 2H)

Biological assessment

Below in conjunction with test case further describe explanation the present invention, but these embodiments be not meant as limiting it is of the invention Range.

Test case 1:

1, measurement of the compounds of this invention to source of people IDO1 protease inhibiting activity

External source of people IDO1 proteinase activity is tested by the following method.

This method is used to measure the compound in the present invention to the inhibiting effect of source of people IDO1 proteinase activity.

One, experimental material and instrument

1, microplate reader (Synergy HT, BIOTEK)

2, tryptophan (T0254-5G, Sigma-Aldrich)

3, catalase derives from cattle liver (C1345-1G, Sigma-Aldrich))

4, methylenum careuleum (M9140-25G, Sigma-Aldrich)

5, L-AA sodium (A7631-25G, Sigma-Aldrich)

6,4- (dimethylamino) benzaldehyde (D2004-25G, Sigma-Aldrich)

7, trichloroacetic acid (T9159-100G, Aigma-Aldrich)

8, IDO1 gene (SC126221, Origene)

9, rosseta (CW0811A, Beijing CoWin Bioscience Co., Ltd.)

10, turbine mixer (6776, Corning)

11, mini board-like centrifuge (mini-p25, ABSON life science equipment)

Two, experimental procedure

The self-control of IDO1 protease

IDO1 gene is transferred in PET30a plasmid by gene clone technology, is then transferred to the Escherichia coli of competence rosseta；In liquid LB (Luria-Bertani) culture medium [according to " Molecular Cloning:A Laboratory guide " (J. Pehanorm Brooker D.W. Russell writes) prepare every liter of culture medium] in amplification culture, collect thallus, ultrasonication affords purifying by hanging column IDO1 protease.

Compound test experiments:

30 times of the enzyme (IDO1) of 100ul is diluted to 3ml with the KPB of 50mM, concentration is the enzyme solutions of 26ng/ul, 96 24ul enzyme solutions are added in the every hole of hole reaction plate (AXYGEN, PCR-96-FLT-C) (hereinafter referred to as reaction plate).Blank well adds Enter 24ul KPB buffer and [preparation (50mM) of KPB buffer: weighs KH with assay balance₂PO₄6.805g being put into 1000ml's Beaker is added deionized water to 900ml with graduated cylinder, adjusts pH to 6.5 with the KOH of 1M, be conducted into the graduated cylinder of 1L, moisturizing is extremely 1L.4 DEG C of storages].Reaction plate be added 1ul compound or DMSO into corresponding reacting hole.Prepare A liquid: taking 250ul 500mM L-AA sodium adds 1437ul KPB, and turbine mixer maximum speed mixes 3 seconds.B liquid: 125ul 10mM color ammonia Acid plus 100000 units of 100ul/ml catalase, add the methylenum careuleum of 5ul 10mM, finally plus 1437ul KPB, are vortexed Mixer maximum speed mixes 3 seconds.1500ul A liquid and 1500ul B liquid are taken, maximum speed mixes 3 seconds on votex.Then This mixed liquor is added in reaction plate with every hole 24ul.Reaction plate is put into board-like centrifuge maximum speed to be centrifuged 15 seconds, is made anti- Liquid is answered all to converge to bottom, oscillator mixes 30 seconds, in constant-temperature incubation case, 37 DEG C, is incubated for 1h.In reaction plate, every hole 10ul30% (W/V) trichloroacetic acid is added, is incubated for 15 minutes for 65 DEG C in incubator.By reaction plate on centrifuge 4700RPM Centrifugation, room temperature, 5 minutes.40ul supernatant is shifted from reaction plate with the volley of rifle fire to corresponding 96 hole test boards (corning, #3599) In.4- (dimethylamino) benzaldehyde/glacial acetic acid solution of 40ul 2% (W/V) is added in every hole, on the oscillator maximum speed, It mixes 1 minute.After incubation at room temperature 2 minutes, the light absorption value at 480nm is read on Synergy HT (BIOTEK).

Compound is measured source of people IDO1 protease inhibiting activity by above test in the present invention, measures IC₅₀Value is shown in Table 1.

Compound inhibits IC to source of people IDO1 proteinase activity in 1 present invention of table₅₀

Conclusion: the compounds of this invention significantly inhibits source of people IDO1 proteinase activity.

The measurement of test case 2, the compounds of this invention IDO protease inhibiting activity intracellular to Hela

The intracellular IDO proteinase activity of Hela is tested by the following method.

This method is used to measure the inhibiting effect of the compound IDO proteinase activity intracellular to Hela in the present invention. (note: HeLa cell strain expresses indoleamine 2,3-dioxygenase (IDO) under the induction of interferon gamma (INF- γ))

One, experimental material and instrument

1, microplate reader (Synergy HT, BIOTEK)

2, tryptophan (T0254-5G, Sigma-Aldrich)

3,4- (dimethylamino) benzaldehyde (D2004-25G, Sigma-Aldrich)

4, trichloroacetic acid (T9159-100G, Aigma-Aldrich)

5, HeLa cell strain (CCL-2, ATCC)

Two, experimental procedure

HeLa cell suspension is produced with Fresh cell culture medium, 100ul cultivating system is added with 10000 cells/wells In 96 porocyte culture plates, 5% 37 DEG C of carbon dioxide is cultivated 24 hours.Supernatant is removed, first 90ul serum-free DMEM is added in every hole High glucose medium；Then every hole is separately added into 10ul and prepares compound (its end in the culture medium of γ containing INF- and tryptophan Concentration are as follows: 100000,10000,1000,100,10,1nM), the training of 96 hole cells is taken out in 48 hours 5% carbon dioxide, 37 DEG C of cultures Supporting supernatant 80ul in plate, into 96 hole round bottom plates, 16ul 30% (W/V) trichloroacetic acid is added in every hole, incubates for 65 DEG C in incubator It educates 25 minutes.By reaction plate, 4700RPM is centrifuged on centrifuge, and 5 minutes.Shifted from reaction plate with the volley of rifle fire 50ul supernatant to In 96 hole flat bottom clear plates, then 4- (dimethylamino) benzaldehyde/glacial acetic acid solution of 50ul 2% (W/V) is added in every hole, It mixes 1 minute on the oscillator.After incubation at room temperature 2 minutes, the extinction at 480nm is read on Synergy HT Reader Value.

Compound IDO protease inhibiting activity intracellular to Hela is measured by above test in the present invention, is surveyed The IC obtained₅₀Value is shown in Table 2.

Compound IDO proteinase activity intracellular to Hela inhibits IC in 2 present invention of table₅₀

Conclusion: the white enzymatic activity of the compounds of this invention IDO Chi intracellular to Hela significantly inhibits.

Pharmacokinetic Evaluation

The pharmacokinetics of test case 3, the embodiment of the present invention 3,6 and 8 compounds is tested

1, it makes a summary

Using SD rat as animal subject, rat intravenous injection is determined using LC/MS/MS method and gives the change of embodiment 3,6 and 8 Drug concentration after conjunction object in different moments blood plasma.The compounds of this invention is studied in the intracorporal pharmacokinetics behavior of rat, is commented Its characteristics of pharmacokinetics of valence.

2, testing program

2.1 test drug

Embodiment 3,6 and 8 compounds

2.2 experimental animal

Healthy adult SD rat 12, half male and half female is purchased from the western Poole-Bi Kai experimental animal in Shanghai Co., Ltd, animal Production licence number: SCXK (Shanghai) 2008-0016.

2.3 drugs are prepared

Appropriate amount of drug is weighed, 0.3ml dimethyl acetamide, which is added, to be made to dissolve, rear that 10%2- hydroxypropyl-β-cyclodextrin is added To final volume, 2.0mg/ml suspension is made in ultrasound.

2.4 administration

SD rat 12, half male and half female is divided into 3 groups；Gastric infusion, administered volume 10ml/ are distinguished after one night of fasting kg。

3, it operates

Gastric infusion group before administration and 0.5 after administration, 1.0,2.0,4.0,6.0,8.0,11.0,24.0h are adopted by eye socket Blood 0.1ml, is placed in heparinised tubes, and 3500rpm is centrifuged 10min separated plasma, saves in -20 DEG C, and 2h is fed after administration.

Untested compound content after measuring different compound gastric infusions with LC/MS/MS method in rat plasma.

4, pharmacokinetic parameter result

The pharmacokinetic parameter of table 3 embodiment of the present invention 3,6 and 8 compounds

Conclusion: in the medicine generation of the compounds of this invention, absorbs well, has apparent medicine for assimilation effect.

Claims (17)

1. a kind of or mixtures thereof logical formula (III) compound represented or its racemic modification form or its officinal salt:

Wherein:

Mixture selected from cis-isomer, transisomer or cis-trans-isomer；

Wherein the ring A is cyclopropyl；

R³And R⁴It is each independently selected from hydrogen atom, C_1-6Alkyl, halogen, wherein the C_1-6Alkyl is optionally further selected from Replaced one or more substituent groups in hydroxyl, halogen；

R⁵Selected from C_1-6Alkyl and amino；

M is 0,1,2；

N is 0,1,2.

2. or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification form or its Officinal salt, wherein R³And R⁴It is each independently selected from hydrogen atom, halogen, C_1-6Alkyl or halogenated C_1-6Alkyl.

3. or mixtures thereof logical formula (III) compound represented according to claim 1 or 2 or its racemic modification form, Or its officinal salt, wherein the halogen is fluorine, chlorine or bromine.

4. or mixtures thereof logical formula (III) compound represented according to claim 1 or 2 or its racemic modification form, Or its officinal salt, wherein the C_1-6Alkyl is C_1~4Alkyl.

5. or mixtures thereof logical formula (III) compound represented according to claim 2 or its racemic modification form or its Officinal salt, wherein the halogenated C_1-6Alkyl is trifluoromethyl.

6. or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification form or its Officinal salt, wherein m is 0.

7. or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification form or its Officinal salt, wherein n is 1.

8. or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification form or its Officinal salt, wherein m is 0 or 1.

9. or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification form or its Officinal salt, wherein n is 1 or 2.

10. a kind of compound is selected from or mixtures thereof compound as described below or its racemic modification form or its officinal salt:

11. a kind of prepare or mixtures thereof logical formula (III) compound represented according to claim 1 or its racemic modification The method of form or its officinal salt, this method comprises:

Open loop obtains logical formula (III) compound to logical formula (VI) compound under alkaline condition；

Wherein:

Ring A, R³~R⁵, m, n it is as defined in claim 1.

12. a kind of or mixtures thereof logical formula (VI) compound represented or its racemic modification form or its officinal salt:

Wherein:

Ring A is cyclopropyl；

R³~R⁵, m, n it is as defined in claim 11.

13. a kind of pharmaceutical composition, general formula described in any one according to claim 1~10 containing therapeutically effective amount (III) or mixtures thereof compound represented or its racemic modification form or its officinal salt and it is one or more pharmaceutically Acceptable carrier, diluent or excipient.

14. logical formula (III) compound represented or its racemic modification described in any one or it is mixed according to claim 1~10 Solvate form or its officinal salt or composition according to claim 13 have in preparation for preventing and/or treating Purposes in the drug of the disease of the pathological characteristics for the tryptophan metabolic pathway that IDO is mediated.

15. purposes according to claim 14, wherein the tryptophan metabolic pathway pathological characteristics mediated with IDO Disease be selected from cancer, myelodysplastic syndrome, Alzheimer disease, autoimmune disease, depression, anxiety disorder, Cataract, mental handicape and AIDS.

16. purposes according to claim 15, wherein the cancer is selected from breast cancer, cervical carcinoma, colon cancer, lung cancer, stomach Cancer, the carcinoma of the rectum, cancer of pancreas, the cancer of the brain, cutaneum carcinoma, carcinoma of mouth, prostate cancer, osteocarcinoma, kidney, bladder cancer, liver cancer, fallopian tubal are swollen Tumor, ovarioncus, peritoneal tumor, IV phase melanoma, glioma, spongioblastoma, the renal tumor of mastoid process, incidence are swollen Tumor, leukaemia, lymthoma and myeloma.

17. purposes according to claim 15, wherein the cancer is selected from hepatocellular carcinoma and non-small cell lung cancer.