CN105440051B - A kind of thiophene pyridine derivate for the treatment of tuberculosis - Google Patents

Abstract

The present invention relates to a kind of thiophene pyridine derivates for the treatment of tuberculosis, are compound of formula I or pharmaceutically acceptable salt, and R1-R5 is as defined herein, which has tubercle bacillus resistant activity.It further include pharmaceutical composition containing a compound of formula I and treatment purposes lungy.

Description

A kind of thiophene pyridine derivate for the treatment of tuberculosis

Technical field

The invention belongs to field of medicinal chemistry, and in particular to the derivative or its optical isomer of a kind of thiophene containing quinoline, Raceme, non-corresponding isomers, pharmaceutically acceptable salt, solvate or its N- oxide and contain above compound Pharmaceutical composition and the compound are used for the purposes of anti-mycobacterium tuberculosis.

Background technique

In global infectious disease statistics, tuberculosis is to be only second to the second lethal infectious disease of AIDS.Although clinical at present The antituberculosis therapy drug of application is numerous, but a line antituberculotic is still the isoniazid for applying many decades, rifampin (Li Fu Mycin class), pyrazinamide and ethambutol.There are clearly disadvantageous in tuberculotherapy for these drugs: (1) drug resistance is tight Weight, and lead to multidrug resistance occurred, or even drug resistant M bacterial strain extensively；(2) treatment cycle is too long, at least requires to advise More than half a year, patient is difficult to adhere to, also promotes the generation of drug resistance for rule medication；(3) there are liver drug enzyme inhibition or inducing action, hold Interaction is generated when easily leading to drug combination, therefore often cannot be with antiviral agent (AIDS patient) drug combination；(4) secondary to make With more, patient is caused to be difficult to adhere to medication.

Due to the prolonged application of existing therapeutic agent and the irregular medication of patient, lead to the tuberculosis bar of patient's body Bacterium generates apparent drug resistance to existing therapeutic agent, or even occurs to a variety of drug resistant multidrug resistances of line antituberculotic Tuberculosis and to the drug resistant extensive drug resistance tuberculosis of all antituberculotics.These drug resistances tubercular is not only because no active drug is treated And case fatality rate is high, and can be transmitted to other people, causes the prevalence of drug resistant M bacterium.The generation of resistant tuberculosis and stream Row, makes tuberculosis become incurable disease again.Claim according to WHO relevant person, lunger will increase to 3,0,000,000 in 10 years from now on More than people, the death of 1,5,000,000 people can only be reduced using existing drug, can the life of the other half people then depends entirely on open Issue the new drug for the treatment of tuberculosis.Therefore, new tubercular drugs action target spot is found, exploitation Newer Antibuberculotics are extremely urgent.

Currently, in the antituberculotic ground, ATP synthetase inhibitors are the completely new mechanism of action of one kind to attract most attention Antituberculotic.Johson & Johnson (WO2004011436A1) discloses a kind of biaryl quinolin analog derivative, and chemical structure is such as Under,

As Beta quinoline (TMC207) has preferable tubercle bacillus resistant activity, but since the compound at present also can only be with Other anti-tubercular drug drug combinations treat tuberculosis, therefore, there is a need to develop active stronger, the higher resistive connection coring of safety Close object.

Summary of the invention

The purpose of the present invention is to provide a kind of compound of formula I and its pharmaceutically acceptable salt or solvate, should Compound belongs to thiophene pyridine derivate, has stronger anti-mycobacterium tuberculosis effect.

On the one hand, in one embodiment, type I compound of the invention or its pharmaceutically-acceptable salts or solvate,

Wherein,

R₁Indicate hydrogen, halogen, hydroxyl, alkoxy, alkyl, cyano, amino or nitro；

R₂For lower straight or branched paraffin containing 0-4 carbon atom；

R₃Indicate phenyl, naphthalene and heterocycle unsubstituted or replaced by one or more substituent groups independent, institute Substitution is stated selected from following groups: halogen, hydroxyl, alkyl, halogenated alkyl, alkoxy, cyano, nitro and amino；

R₄、R₅ When for two groups independent, it is respectively selected from hydrogen, halogenated alkyl, hydroxyl, alkyl, alkoxy, acyl Base, sulfonyl, Ar, Ar- alkyl, Het and Het-alkyl substituent group；

Work as R₃And R₄For connect together group when, be saturated or unsaturated containing 1-4 selected from that can be formed with nitrogen-atoms The group of heteroatomic 4-7 member ring.

In one embodiment, type I compound of the invention or its pharmaceutically acceptable salt or solvate,

Wherein,

R1 indicates hydrogen, halogen, cyano, alkoxy and alkyl；

R₂For methyl, ethyl or propyl

R₃Indicate phenyl, naphthalene and heterocycle unsubstituted or replaced by one or more substituent groups independent, institute Substituent group is stated to replace selected from halogen, hydroxyl, alkoxy and alkyl；

R₄、R₅For two independent groups, it is respectively selected from hydrogen, halogenated alkyl, hydroxyl, alkyl, alkoxy, acyl group, sulfonyl Base, Ar, Ar- alkyl, Het and Het-alkyl substituent group；

In preferred embodiments, type I compound of the invention or its pharmaceutically acceptable salt or solvate,

Wherein,

R₁Indicate halogen；

R₂For methyl;

R₃Indicate phenyl, naphthalene and heterocycle unsubstituted or replaced by one or more substituent groups independent, institute Substituent group is stated to replace selected from halogen, alkoxy and alkyl；

R₄、R₅ For two independent groups, it is respectively selected from hydrogen, halogenated alkyl, alkyl and acyl substituent.

Preferably, compound of formula I of the invention or its pharmaceutically acceptable salt are selected from following compound:

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (naphthalene -1- base) -1- phenyl -2- Butanol；

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3- fluorophenyl) -1- phenyl -2- Butanol；

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3- methoxyphenyl) -1- benzene Base -2- butanol；

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3,5- difluorophenyl) -1- benzene Base -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -1,2- diphenyl -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -1- phenyl -2-(3- methyl Phenyl) -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(3- fluorophenyl) -1- benzene Base -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -2-(3,5- difluorophenyl) -4- (dimethylamino) - 1- phenyl -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(naphthalene -1- base) -1- benzene Base -2- butanol；

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(3- methoxybenzene Base) -1- phenyl -2- butanol；

N- (4- (bromo- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -3- hydroxyl -3- (3- methoxyphenyl) -4- benzene Base butyl)-N- methylacetamide.

Compound of formula I of the invention or its pharmaceutically acceptable salt or solvate, the pharmaceutically acceptable salt Including but not limited to hydrochloride, hydrobromate, sulfate, mesylate, tosilate, maleate, fumarate, flat Peach hydrochlorate, citrate or tartrate；The solvate of the formation such as the solvated compounds and water, alcohol, ketone, as hydrate, Methanol solvate, ethanolates etc., wherein R₁、R₂、R₃、R₄、R₅It is defined as above.

Compound of formula I of the invention or its pharmaceutically acceptable salt, the type I compound further include its optics

Isomers, raceme, non-corresponding isomers or N oxygen compound.The optical isomer can pass through conventional hand Property removes point method or Chiral Synthesis obtains, wherein R₁、R₂、R₃、R₄、R₅It is defined as above.

On the other hand, the object of the invention is also to provide a kind of pharmaceutical compositions, including containing the change of above-mentioned formula I of the invention It closes object or its optical isomer, pharmaceutically acceptable salt or solvate, and optional one or more pharmaceutically may be used The auxiliary material or carrier of receiving, wherein R₁、R₂、R₃、R₄、R₅It is defined as above.

It yet still another aspect, this hair is designed to provide a kind of therapeutical uses, i.e., type I compound of the invention or its optics Isomers, raceme, non-corresponding isomers, pharmaceutically acceptable salt or solvate manufacture treatment and/or prevent by Disease caused by tubercle bacillus or the utilization in disorder agent；Namely a kind for the treatment of and/or prevention are as caused by tubercle bacillus The method of disease or illness, compound of formula I (wherein, the R including applying effective dose to tuberculosis patient or subject₁、R₂、 R₃、R₄、R₅It is defined as above) or its optical isomer, raceme, non-corresponding isomers, pharmaceutically acceptable salt or solvent Object is closed, described as tuberculosis is the pulmonary tuberculosis as caused by tubercle bacillus.

The various terms and phrase that the present invention uses have well known to a person skilled in the art general senses, even if such as

This, the present invention is described in more detail and explains to these terms and phrase again, and the term and phrase referred to is such as Have inconsistent with common art-recognized meanings, is subject to meaning expressed by the present invention.Here is the definition of certain terms, these definition are suitable For term used in the application the whole instruction, unless otherwise indicated in concrete condition.

In the context of the present invention, discribed such as in general formula compound or particular compound, some atomic group can To connect several hydrogen atoms, the atomic group is made to can satisfy the requirement of chemical valence, although the hydrogen atom on corresponding atomic group It is depicted without in structural formula.

" alkyl " is the hydrocarbon group of the linear chain or branched chain of 1-6 carbon atom；Saturated rings containing 3-6 carbon atom Shape group；The group of linear chain or branched chain containing 1-6 carbon atom and the saturated rings containing 3-6 carbon atom, the hydrogen on the alkyl It can be replaced by hydroxyl or halogen；

" alkoxy " refers to alkyl-O-, and wherein alkyl is as hereinbefore defined；

" halogen " be selected from fluorine, chlorine, bromine, iodine substituent group；

" halogenated alkyl " is the linear chain or branched chain saturated hydrocarbyl with 1-6 carbon atom or the ring with 3-6 carbon atom Shape saturated hydrocarbyl, wherein one or more carbon atoms replace by one or to a halogen atom；

" Ar " be selected from phenyl, naphthalene, tetralyl carbocyclic ring, each carbocyclic ring optionally takes by 1,2 or 3 substituent group Generation, each substituent group solely select from hydroxyl, halogen, cyano, nitro, amino, list or dialkyl amido, alkyl, halogenated alkyl, Alkoxy, halogenated alkoxy, carboxyl, alkoxy carbonyl, amino carbonyl, morpholinyl and list or dialkyl amino carbonyl；

" Het " be selected from N- Phenoxypiperidines base, piperidyl, pyrrole radicals, pyrazolyl, imidazole radicals, furyl, thienyl, Oxazole, isoxazole, pyridyl group, pyrimidine radicals, pyrazinyl and pyridazinyl monocyclic heterocycles；Or for selected from quinolyl, quinazolyl, Yin Diindyl base, benzimidazolyl, phenylpropyl alcohol oxazolyl, benzo isoxazolyl, benzothiazole, benzisothia oxazolyl, benzofuran, benzo thiophene Pheno base, each monocycle or bicyclic heterocycle are optionally selected from the substitution of halogen, hydroxyl, alkyl, alkoxy or Ar- carbonyl by 1,2 or 3 Base replaces；

" subject " can refer to patient or receive type I compound of the present invention or its pharmaceutical composition to treat and/or in advance Prevent the animal of disease or illness of the present invention, especially mammal, such as people, dog, monkey, ox, mouse etc..

" disease and/or illness " refers to a kind of physical condition of the subject, the physical condition and disease of the present invention Disease and/or illness are related.

As described herein, as not specified, " % " refers to w/w percentage, especially in description solid matter In the case of.Certainly, when describing liquid substance, being somebody's turn to do " % " can refer to that weight/volume percent (is dissolved in solid the feelings of liquid Shape), or can refer to volume/volume percentage (situation that liquid is dissolved in liquid).

As described herein, " pharmaceutically acceptable " or " pharmaceutical " that is used interchangeably with it, such as describing It indicates that the salt is not subjected on subject physiologic still when " pharmaceutically acceptable salt ", but also can refer to pharmaceutically have use The synthetic of value, such as it is formed by when carrying out chiral resolution the salt as intermediate, although the salt of this intermediate Can not directly give subject, but the salt can for or final product of the present invention in work.

In another aspect, preparing type I compound and its optical isomer or its medicine the object of the present invention is to provide a kind of The method of acceptable salt on, method includes the following steps:

1) III formula of formula, II compound is made by the reaction of Wei Er David Smail, and Wei Er David Smail reagent is in low temperature Under the conditions of by DMF instill phosphorus oxychloride stirring be made, the cryogenic conditions be preferably -20 DEG C to 0 DEG C, II compound of formula be commercially available production Product, R₁It is identical as the definition in above-mentioned type I compound；

2) IV formula of formula, III compound occurs in anhydrous THF with phosphonoacetate under alkaline condition Witting-Horner reaction is made.Alkali used in the reaction is preferably HMDSNa, HMDSLi, potassium tert-butoxide and DBU etc., R₁ It is identical as the definition in above-mentioned type I compound；

3) V formula of formula, IV compound restores in tetrahydrofuran is made, which is preferably diisobutyl hydrogen Change the reducing agents such as aluminium, Lithium Aluminium Hydride, red aluminum solution, R₁It is identical as the definition in above-mentioned type I compound；

4) VI formula of formula, V compound back flow reaction in the corresponding alcoholic solution of sodium alkoxide is made, R₁、R₂With above-mentioned formula Definition in I compound is identical；

5) VII formula of formula, VI compound is made by summer Price asymmetric epoxidation reaction, in the reaction Oxidant is preferably tert-Butanol peroxide, R₁ 、R₂It is identical as the definition in above-mentioned type I compound；

6) by grignard reagent, the open loop under the catalysis of cuprous salt obtains VIII formula, VII compound, and the cuprous salt is excellent It is selected as cuprous iodide, cuprous bromide and cuprous cyanide, R₁ 、R₂It is identical as the definition in above-mentioned type I compound.

7) Ⅸ formula of formula, VII compound is after sodium periodate oxidation is at corresponding aldehyde, then and R₃Format examination Agent reaction is made, wherein R₁、R₂ 、R₃It is identical as the definition in above-mentioned type I compound；

8) Ⅹ formula of formula, Ⅸ compound oxidation obtains, which preferably uses Jones reagent oxidation, wears This-Martin reagent oxidation, it is polite oxidation the methods of, R₁、R₂ 、R₃It is identical as the definition in above-mentioned type I compound；

9) Ⅺ formula, Ⅹ compound is reacted with allyl grignard reagent at low temperature is made, and the reaction temperature is optional It selects at -100 DEG C -0 DEG C, allyl grignard reagent is preferably allylic bromination magnesium, allylmgcl and allyl magnesium iodide, R₁、R₂ 、R₃It is identical as the definition in above-mentioned type I compound；

10) Ⅻ formula of formula, Ⅺ compound double bond oxidation scission generates aldehyde and is made, and the method for oxidation in the reaction is excellent It is selected as osmium tetroxide and increases sodium iodate and ozone oxidation, R₁、R₂、R₃It is identical as the definition in above-mentioned type I compound.

11) I formula compound by Ⅻ compound and contains R₄、R₅The amine of substituent group occurs reductive amination process and obtains, the reaction Reducing agent is preferably sodium borohydride, sodium cyanoborohydride and acetic acid sodium borohydride；R₁、R₂、 R₃、R₄、R₅With above-mentioned type I compound In definition it is identical.

Specific synthetic route of the invention is as follows:

Wherein R₁、 R₂、R₃、R₄And R₅It is identical as the definition in above-mentioned type I compound.

The compound of formula I of the invention or its pharmaceutical salt can be used alone, or in the medicinal auxiliary material of addition, carrier Or excipient forms the form of pharmaceutical composition and uses, when in the form of pharmaceutical composition in use, usually will effectively measure Type I compound of the invention or its officinal salt or solvate and one or more pharmaceutically acceptable auxiliaries, carrier or diluent are mixed Suitable dosage form is made in conjunction, adopts and with the conventional methods in the field mixes component, granulation, compresses or dissolve.

The pharmaceutical composition for the compound that the present invention contains formula I can be administered selected from any mode of following aspect: oral, Spraying sucking, rectally, nasal-cavity administration, vagina administration, local administration, parenterai administration such as subcutaneous, vein, muscle, peritonaeum In interior, intrathecal, intra-ventricle, breastbone or intracranial injection or input or by a kind of external reservoir medication, wherein preferably take orally, In intramuscular injection, peritonaeum or intravenous administration mode.

Compound of formula I of the invention can be administered in a unit containing its pharmaceutical composition.Form of administration can To be liquid dosage form, solid dosage forms.Liquid dosage form can be true solution, colloidal type, particulate formulations, emulsion dosage form, mixed dosage form. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, mixed agent, emulsion, granule, suppository, freeze-drying Powder needle, inclusion compound, implants, patch, liniment etc..

Common pharmaceutic adjuvant can also be contained in pharmaceutical composition of the invention, described pharmaceutical acceptable carrier includes but unlimited In: ion-exchanger, aluminium oxide, aluminum stearate, lecithin, haemocyanin such as human albumin, buffer substance such as phosphate, Glycerol, sorbic acid, potassium sorbate, the partial glyceride mixtures of saturated vegetable fatty acid, water, salt or electrolyte, such as sulfuric acid fish Protamine, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, cabosil, magnesium trisilicate, polyvinylpyrrolidone are fine Tie up plain substance, polyethylene glycol, sodium carboxymethylcellulose, polyacrylate, beeswax, wool grease etc..Auxiliary material is in pharmaceutical composition Content can be 1% -98% weight.

Auxiliary material used in oral tablet and capsule such as includes adhesive, such as syrup, Arabic gum, sorbierite, tragacanth or poly- Vinylpyrrolidone, filler, such as lactose, sucrose, cornstarch, calcium phosphate, sorbierite, amion acetic acid, lubricant is such as stearic Sour magnesium, talcum, polyethylene glycol, tripoli, disintegrating agent, such as potato starch, low-substituted hydroxypropyl methylcellulose, carboxymethyl cellulose Sodium, cross-linked polyvinylpyrrolidone etc. or acceptable dibutyl phthalate, such as laurel sodium alkoxide sulfate.Tablet can be in pharmaceutics Well known method coating.

The suspension of water and oil, solution, emulsion, syrup or elixir can be made in oral solution, and dry product can also be made, and use Preceding supplement water or other suitable mediums.This liquid preparation may include conventional additive, such as suspending agent, and sorbierite is fine Tie up plain methyl ether, dextrose syrup, gel, hydroxyethyl cellulose, carboxymethyl cellulose, aluminium stearate gel, the edible oil of hydrogenation Rouge, emulsifier, such as lecithin, sorbitan mono-oleate, Arabic gum；Or nonaqueous carrier (may include edible oil), such as Apricot kernel oil, grease such as glycerol, ethylene glycol or ethyl alcohol；Preservative, such as methyl p-hydroxybenzoate or propyl ester, sorbic acid.As needed Flavoring agent or colorant can be added.

Suppository may include conventional suppository base, such as cocoa butter or other glyceride.

Parenteral, usually injection, usually by the compound of the present invention and injection supplementary material and/or solvent system At.The solvent first choice is water.The compound of the present invention is dissolved in solvent and forms solution, or be made in a solvent suspend it is molten Liquid, can be soluble in water by the compound of the present invention when injection solution is made, and is fitted into sealed bottle or ampoule after filtration sterilization.

When topical application, the compounds of this invention can be made ointment appropriate, the form of lotion or creme, Middle active constituent is suspended or dissolved in one or more carriers.The carrier that wherein ointment formulation can be used includes but unlimited In: mineral oil, Albolene, albolene, propylene glycol, polyethylene glycol oxide, polypropylene oxide, emulsifying wax and water；Lotion and Carrier workable for creme includes but is not limited to: mineral oil, sorbitan monostearate, polysorbate60, cetyl ester wax, Hexadecene is fragrant and mellow, 2- octyldodecanol, benzyl alcohol and water.

In said medicine preparation, formula (I) compound there are about 0.1 that concentration should be the total weight of the mixture 99.5%, preferably from about 0.5 95% (weight).

Said medicine preparation also can further include other pharmaceutical activity chemical combination other than the compound comprising formula (I) Object.

- as, it has proved that advantageously no matter human body medicine or in veterinary drug, the compound of the present invention Be within administration total amount every 24 hours about 0.5-500mg, preferably l-100mg, if applicable, single dose is administered several times, with Achieve the effect that required.Amount in single dose containing the compound of the present invention is preferably from about l-80mg, more preferably l-50mg, but The type and weight, the property of disease and severity, preparation for the treatment of object can not also be depended on according to above-mentioned dosage Type and drug administration mode and dosage period or time interval.

Specific embodiment

Following embodiment is the preferred embodiments of the invention, should not be construed as constituting any restrictions to the present invention.

The fusing point of compound is measured by SGW X-4 type micro-meldometer, the non-calibration of thermometer.Mass spectrum is by SHIMADZU LCMS-2020 measurement.NMR is by Agilent-NMR-vnmrs400 and Agilent-NMR-vnmrs600

In the preparation process of each compound, the absolute stereochemistry of asymmetric carbon atom is not determined experimentally.? In this case, steric compounds configuration is not considered.

The preparation of intermediate

Embodiment 1

The preparation of intermediate III -1

Phosphorus oxychloride 141ml(1.54mol) in 1000ml single port bottle, it is cooled to 0^oDMF 58ml is slowly added dropwise in C Maintenance 0 is added dropwise in (0.77mol)^oC reacts 30min, is warming up to room temperature, is slowly added to 2- acetylamino 5- bromothiophene by several times 29.4g(0.193mol), it stirs evenly, is warming up to 80^oC reacts 1.5h, and reaction solution is cooled to room temperature, has been poured slowly into In 1200g ice water, stir 30 minutes, after be extracted with ethyl acetate, liquid separation, organic phase saturated common salt water washing, anhydrous slufuric acid Sodium is dry, after concentration crude product 300ml ethyl alcohol recrystallization, obtain 40.1g beige solid intermediate III -1, yield: 75.1%。

Embodiment 2

The preparation of intermediate IV -1

Phosphonoacetate 39.3g(0.161mol), about 50mlTHF is added, is cooled to 0^oC be slowly added dropwise into HMDSNa solution 81ml(0.161mol), it is warming up to room temperature after being added dropwise and is stirred to react 30min, which is slowly added to Into the THF(250ml of 40.0g(0.146mol) intermediate III -1) suspension solution, it is stirred to react about 2.5h, TLC inspection at room temperature It is complete to survey fundamental reaction.Water 100ml quenching reaction is added, is extracted with ethyl acetate, saturated common salt water washing, anhydrous sodium sulfate is dry Dry, gained crude product column chromatographic purifying, mobile phase solvent proportion is ethyl acetate: petroleum ether=1:5 finally obtains 38.6g light gray Color solid intermediate IV -1, yield: 76.8 %.

Embodiment 3

The preparation of intermediate V -1

33.2g(0.96mol) dissolution of 300ml methylene chloride, ice-water bath is added in 1000ml single port bottle in intermediate Ⅸ -1 Under the toluene solution 240ml(1M/L, 0.24mol of diisobutyl aluminium hydride is slowly added dropwise), room temperature reaction is warming up to after dripping 1.5h, TLC detect fully reacting.Reaction is added after about 150ml ice water is quenched and is poured slowly into the 2N hydrochloric acid solution containing 1L ice cube In, the liquid separation after its completely clarification, water layer is extracted with dichloromethane twice, and combined organic phase is washed with saturated common salt and washed, Anhydrous sodium sulfate is dry, and crude product ethyl acetate: petroleum ether=1:6 is recrystallized, and obtains 26.3g dark yellow solid intermediate V -1, yield: 90.3%.

Embodiment 4

The preparation of intermediate VI -1

Metallic sodium 14g(0.605mol), it is slowly added to after being cut into small pieces in the 1000ml single port bottle equipped with 500ml methanol, Stirring condensation to sodium block dissolution completely, by several times be added 26.3g(0.086mol) intermediate V -1, after be warming up to 80^oC, stirring are anti- It answers (about 16h), TLC detection reaction.After distillation for removing methanol plus water 300ml dilution, after be extracted with dichloromethane, liquid separation is organic Layer saturated salt solution washing, anhydrous sodium sulfate is dry, and gained crude product methylene chloride: petroleum ether=1:2 is recrystallized, and is obtained 18.6g light yellow solid intermediate VI -1, yield: 71.8%.

Embodiment 5

The preparation of intermediate VII -1

Isopropyl titanate 2.57g(0.0091mol) in 500ml there-necked flask, it is molten that about 150ml dry methylene chloride is added Solution, and dry 4A molecular sieve 20g is added, it is cooled to -20^oL- (+) tartaric acid 3.18g(0.0136mol is slowly added dropwise in C) Dichloromethane solution (about 30ml) after being added dropwise, keeps temperature and is stirred to react 30min.6.8g is added by several times afterwards (0.0226mol) intermediate VI -1, continues to be stirred to react 40min；Anhydrous tert-butyl hydroperoxide well prepared in advance is added afterwards Toluene solution 56ml(about 4M/L, 0.226mol), still -19 ~ -23^o3.5h is reacted under C, TLC detection raw material almost converts Completely.20% sodium hydroxide solution about 100ml stirring 1.5h is added and is raised to room temperature naturally, filters, liquid separation, each 150ml of water layer Methylene chloride extracts 2 times, and saturated common salt is washed after organic layer merges, and anhydrous sodium sulfate is dry, crude product column chromatographic purifying, flowing Phase solvent proportion be ethyl acetate: petroleum ether=3:10, after obtain 5.5g light yellow solid intermediate VII -1, yield: 77.0%.

Embodiment 6

The preparation of intermediate VIII -1

7.87g(0.328mol) magnesium chips is added in dry 250ml there-necked flask, and under nitrogen protection, dry THF is added 80ml, and 50mg iodine is added and stirs evenly, is slowly added dropwise into bromobenzene 17ml(0.164mol), to its initiation reaction after again It is heated to 55 ~ 60^oC keeps the temperature 1h, is placed at room temperature for spare.Carefully weigh cuprous cyanide 8.9g(0.0987mol) it is placed in 500ml single port It is rear that dry THF 150ml is added in bottle, -40 are cooled to acetonitrile-liquid nitrogen system^oC under nitrogen protection, is slowly added dropwise and is cooled to The spare bromobenzene grignard reagent of room temperature maintains -40 after being added dropwise^oC is simultaneously stirred to react 1h.5.2g(0.0164mol) is intermediate Body VII -1 is dissolved in 30mlTHF and is slowly added dropwise into reaction system, continues -30 ~ -40^o2h is reacted under C, TLC detection reaction is originally Completely, 80ml aqueous ammonium chloride solution quenching reaction is added in conversion, and ethyl acetate extraction is added, filters, liquid separation, water layer is respectively used 100ml ethyl acetate extracts 2 times, merges organic layer and is washed with saturated common salt, anhydrous sodium sulfate is dry, gained crude product column layer Analysis purifying, mobile phase solvent proportion is ethyl acetate: petroleum ether=1:2 finally obtains the faint yellow oily intermediate VIII -1 of 4.0g, Yield: 61.4%.

The preparation of intermediate VIII -2:

Using 2- acetamido -5- chlorothiophene as raw material, reference implementation example 1,2,3,4,5,6, intermediate III -1, IV -1, V - 1, VI -1, VII -1 and VIII -1 preparation approach, intermediate VIII -2 can be made.

Embodiment 7

The preparation of intermediate Ⅸ -1:

1.5g(0.0038mol) methylene chloride 20ml stirring and dissolving is added in 100ml single port bottle in intermediate VIII -1, after 20% sodium metaperiodate silica gel reagent 18g(0.027mol is added), water 0.1ml is stirred at room temperature reaction 1.5h, TLC detection and has reacted Entirely, it filters, anhydrous sodium sulfate is dry, and decompressing and extracting is spare.

Weigh 0.82g(0.034mol) magnesium rod and it is cut into bits shape, and add it in bis- mouthfuls of bottles of dry 100ml, add Enter a small amount of iodine and stir evenly, under nitrogen protection, dry THF 30ml is added, is warming up to 50^oIt is slowly added dropwise after C into bromonaphthalene 5.3ml(0.038mol), to its initiation and after completion of the reaction 55 ~ 60^o1h is kept the temperature under C, is cooled to room temperature, and is cold with ice-water bath But to 0^oC under nitrogen protection, is slowly added dropwise the THF solution (about 10ml) of spare compound, keeps 0^oC reacts 2h, and TLC detection is anti- It should convert substantially completely.30ml aqueous ammonium chloride solution quenching reaction is added, is extracted with ethyl acetate, water layer is each respectively again to use 50ml Ethyl acetate extracts 2 times, and saturated common salt is washed after organic layer merges, and anhydrous sodium sulfate is dry, and crude product column chromatographs pure after concentration Change, mobile phase solvent proportion is ethyl acetate: petroleum ether=1:10 obtains 1.45g light yellow solid intermediate Ⅸ -1, yield: 78.1%。

The preparation of Ⅸ -2 to Ⅸ -11 intermediate of formula:

Referring to the preparation method of Ⅸ -1 compound of intermediate formula of embodiment 7, it is with the compound of formula VIII -1 and VIII -2 respectively Raw material reacts to obtain the Formula IX compound intermediate (abbreviation Formula IX intermediate) of the following table 1 with corresponding grignard reagent.

Embodiment 8

The preparation of Ⅹ -1 compound intermediate of formula (referred to as " intermediate Ⅸ -1 ")

Weighing 1.2g(0.0024mol) intermediate Ⅸ -1 is in 100ml there-necked flask, addition methylene chloride 30ml dissolution, after The Dess-Martin reagent 2.54g(0.006mol of brand-new is added), after be stirred to react 3h at room temperature, TLC detection reaction has converted Entirely, 20ml saturated aqueous sodium thiosulfate quenching reaction, filtering is added, filtrate is stirred with saturated sodium bicarbonate aqueous solution, point Liquid, water layer are respectively extracted with dichloromethane secondary, combined organic layer and use saturated common salt water washing again, and anhydrous sodium sulfate is dry, slightly Product column chromatographic purifying, mobile phase solvent proportion is ethyl acetate: petroleum ether=1:25 finally obtains among 900mg white solid Body Ⅹ -1, yield: 76.9%.

Embodiment 9

The preparation of intermediate Ⅺ -1

Under nitrogen protection, the diethyl ether solution 18ml(1.0M/L, 0.0184mol of allylic bromination magnesium) is injected into 100ml In single port bottle, -78 are cooled to acetone-liquid nitrogen system^oC is slowly added dropwise into 900mg(0.00184mol) intermediate Ⅹ -1 8ml THF solution, it is rear to maintain -80^oC to -68^o1.5h is reacted under C, TLC detects fully reacting.20ml aqueous ammonium chloride solution is added Quenching reaction, after be extracted with ethyl acetate, water layer respectively use 30ml ethyl acetate extraction three times, organic layer merging uses saturated salt solution Washing, anhydrous sodium sulfate is dry, crude product column chromatographic purifying, and mobile phase solvent proportion is ethyl acetate: petroleum ether=1:20 obtains 762mg light yellow solid intermediate Ⅺ -1, yield: 78.0%.

Embodiment 10

The preparation of intermediate Ⅻ -1:

595mg(1.12mmol) intermediate compound I 1 is placed in 50ml single port bottle, with mixed solvent 6ml(dioxane: water=3: 1) dissolve, 2,6- lutidines 261ul(2.24mmol be added afterwards), it is slowly added dropwise into osmium tetroxide solution 214ul(0. 16mmol), sodium metaperiodate 980mg(4.6mmol is then added), it is stirred to react 2.5h, TLC detection reaction at room temperature.2ml is added Water quenching reaction, it is rear to filter, it is extracted with ethyl acetate, water layer is respectively extracted 2 times with 5ml ethyl acetate again, organic layer saturation food Salt water washing, anhydrous sodium sulfate is dry, and revolving removes solvent, drains, obtains 450mg light brown oil product Intermediate Ⅻ -1, nothing Investment need to be purified to react in next step.Yield: 75.6%

The preparation of intermediate Ⅻ -2 to Ⅻ -10:

Referring to the preparation method of the intermediate of embodiment 8,9,10, respectively under formula Ⅸ -2 to Ⅸ -10 intermediate of formula preparation Ⅻ intermediate of formula (formula Ⅻ -2 to Ⅻ -11) in table 2.

The preparation of target compound:

The preparation of 11 chemical compounds I -1, I -2 of embodiment:

586mg(0.0011mol) compound Ⅻ -1 is dissolved with 15ml methanol, rear that dimethylamine hydrochloride 911mg is added (0.011mol), sodium acetate 1.673g(0.0123mol), sodium cyanoborohydride 347mg(0.0055mol), reaction is at room temperature It is stirred to react overnight (about 16h), TLC and LC-MS detection raw material conversion are complete.Vacuum distillation removes most of methanol, adds water 20ml dilution, is extracted with dichloromethane, liquid separation, and water layer uses methylene chloride 20ml to extract 3 times again, combined organic layer saturation food Salt water washing, anhydrous sodium sulfate is dry, gained crude product column chromatographic purifying, and mobile phase solvent proportion is acetone: petroleum ether=3: 20, two non-corresponding isomers are obtained, the component first come out is labeled as chemical compounds I -1, total 195mg, yield: 31%.It comes out afterwards Component is labeled as the total 20mg of chemical compounds I -2, yield: 3.4%.

Embodiment 12 is made referring to 10 prepare compound I -1 of embodiment, I -2 method by Ⅻ compound of intermediate formula respectively Each compound in standby the following table 3 out, structure see the table below:

The fusing point and nuclear magnetic data of Formulas I -1 to -21 compound of Formulas I are shown in Table 4, and table 4 is Formulas I -1 to -21 compound of Formulas I Fusing point and nuclear magnetic data.

The compound of the present invention Tuberculosis in vitro core bacillus active (MIC) detection

Detection content

1) Killing Mycobacterium Tuberculosis active testing (mycobacterium bovis GFP-BCG bacterial strain, M. tuberculosis mycobacteria H37Rv)；

2) Killing Mycobacterium Tuberculosis clinical separation strain active testing (is clinically separated 3 plants of sensitive strain, is clinically separated persister 5 Strain)

Detection method

1) type strain, clinical separation strain of M. tuberculosis mycobacteria and clinical drug-resistant strain (Chinese CDC tuberculosis laboratory The bacterial strain of preservation) it is detected using microplate Alamar Blue assay, it tests in Disease Control and Prevention in China Center tuberculosis Reference Lab is completed.It is such as not specified, the working concentration of drug is set as 25.6ug/ml and starts, and 2 times of gradients are dilute It releases.Blank control group is DMSO.

2) all experiments are repeated three times.

Detecting step

One, the preparation of stock sample solution (1.28 mg/mL)

The equal precise of each sample.According to sample message, the DMSO of certain volume is added separately in various kinds quality control, is surpassed Sound hydrotropy is formulated as the stock sample solution of 1.28mg/ml respectively.

Two, the preparation of chemicals dilating version

Stock sample solution is subjected to 2 times of gradient dilutions, packing to 96 orifice plates, each hole 200ul with DMSO respectively.Sample Concentration are as follows:

1.28mg/ml,0.64mg/ml,0.32mg/ml,0.16mg/ml,0.08mg/ml,0.04mg/ml,0.02mg/ ml,0.01mg/ml

Three, the preparation of bacteria suspension

BCG is referring to above method.

The bacteria suspension preparation method of M. tuberculosis mycobacteria is added in the Roche culture tube containing mycobacteria as follows Appropriate amount of fluid culture medium is gently blown and beaten with suction pipe, is drawn bacterium solution and is set in mill tube, after sufficiently grinding bacterium on vortice, stands 15 Min, with opacity tube, (1 Maxwell, dense bacterium is 3 × 10⁸) carry out than turbid, dense final bacterium is 3 × 10⁶

Four, it is loaded

50ul culture medium is added in the healthy and free from worry each hole of 96 orifice plate of Bacteria Culture (flat), then with 96 hole microwell plate reproducers The compound of 2ul is shifted to culture plate.Then 48 hole μ L/ of bacterium solution diluted is added into 96 well culture plates.DMSO is most Final concentration of 2%, it is shown in Table 5.It after adding bacteria suspension, closes the lid, is sealed plank with adhesive tape, set 37 DEG C of incubators and carry out Culture.

Five, testing results

The Resazurin solution of configured in advance 0.1mg/ml, filtration sterilization.

After being incubated for 7 days, 20ul Resazurin is added, the variation of well color is observed after three days, blue indicates chemical combination Object inhibits bacterium growth, pink colour to indicate that compound is inactive.According to taking pictures as a result, the bacterial plaque size that detects by an unaided eye to plank, into The description of strain growth situation in each drug hole of row, minimum inhibitory concentration MIC value is defined as minimum contained in asepsis growth hole Antibacterials concentration (MIC90%).It the results are shown in Table 6, table 6 is human-like mycobacterium tuberculosis Activity Results (ug/ml).

Mycobacterium bovis active testing

Experimental method

PUV3583c-gfp plasmid is carried in this experiment BCG JP15 bacterial strain used, which can answering with BCG Constitutive expression GFP processed reflects drug to the inhibition situation of bacterium, compared with based on light absorption value with the GFP fluorescence intensity detected The method of detection has the advantages that sensitivity is higher, signal window is wider, has compared with decoration methods such as Alamar Blue, MTT and is not necessarily to The advantage that secondary sample-adding, step simplify.The bacterial strain can test in-house operation in bio-safety second level.Anti- BCG Activity determination mould Type is according to Clinical and Laboratory Standards

The modification of Institute Guide Book M24-A method, carries out in bio-safety second level laboratory.Firstly, will Mycobacterium bovis BCG attenuated strain BCG JP15 is seeded in 7H9 culture medium, and 60rpm cultivates 7 days to right in 37 DEG C of constant-temperature tables Number growth medium.Being diluted to OD600=0.025(with fresh 7H9 culture medium, corresponding ~ 106 CFU/mL bacterium are dense at this time), as Bacteria suspension.Each untested compound is dissolved in dimethyl sulfoxide (DMSO) and carries out 2 times of gradient dilutions, draws 1 μ L untested compound Solution is added in 96 each holes of hole microwell plate, then draws 80 μ L of bacteria suspension with multichannel pipettor and is added in 96 each holes of hole microwell plate. Negative control is DMSO.Each test board is closed with Parafilm sealed membrane, is placed in 37 DEG C of constant incubators after being incubated for 96 hours, It is detected using fluorescence microplate reader (Perkin Elmer Envision 2100) and records each hole fluorescent value, excitation wavelength is 480nm, launch wavelength 535nm.BCG growth is suppressed the hole institute of 90% or more (fluorescent value is lower than 90% or more blank control group) Corresponding final compound concentration is defined as the compound to the minimum inhibitory concentration (MIC) of BCG.

Experimental result is shown in Table 7.

。

Claims (5)

1. type I compound or its pharmaceutically acceptable salt,

Wherein,

R₁Indicate halogen；

R₂For methyl, ethyl or propyl

R₃Indicate that phenyl and naphthalene unsubstituted or replaced by one or more substituent groups independent, the substituent group are selected from Halogen, hydroxyl, alkoxy and alkyl, wherein the alkyl in alkyl and alkoxy is the linear chain or branched chain of 1-6 carbon atom Hydrocarbon group；

R₄、R₅For two independent groups, it is respectively selected from hydrogen and alkyl, the alkyl is the linear chain or branched chain of 1-6 carbon atom Hydrocarbon group.

2. the compound of general formula I according to claim 1 or its pharmaceutically acceptable salt, wherein

R₁Indicate bromine；

R₂For methyl；

R₃Indicate that phenyl and naphthalene unsubstituted or replaced by one or more substituent groups independent, the substituent group are selected from Halogen, alkoxy and alkyl, wherein the alkyl in alkyl and alkoxy is methyl；

R₄、R₅ For two independent groups, it is selected from hydrogen and methyl.

3. compound of formula I according to claim 1, it is characterised in that compound selected from the following:

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (naphthalene -1- base) -1- phenyl -2- butanol

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3- fluorophenyl) -1- phenyl -2- butanol

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3- methoxyphenyl) -1- phenyl -2- Butanol

1- (bromo- 6- methoxyl group [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2- (3,5- difluorophenyl) -1- phenyl -2- Butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -1,2- diphenyl -2- butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -1- phenyl -2-(3- methylbenzene Base) -2- butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(3- fluorophenyl) -1- phenyl - 2- butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -2-(3,5- difluorophenyl) -4- (dimethylamino) -1- Phenyl -2- butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(naphthalene -1- base) -1- phenyl - 2- butanol

1- (chloro- 6- methoxythiophene [2,3-b] pyridine -5- base of 2-) -4- (dimethylamino) -2-(3- methoxyphenyl) -1- Phenyl -2- butanol.

4. a kind of pharmaceutical composition, including compound described in claim 1-3 or pharmaceutically acceptable salt and medicine Acceptable auxiliary material on.

5. claim 1-3 any compound or pharmaceutically acceptable salt treat or prevent mycobacterium tuberculosis infection in preparation Utilization in caused disease or disorder agent.