CN101817834B - Pyrazole derivatives and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the technical field of antifungal action medicine and provides pyrazole derivatives of the structure in the general formula I and pharmaceutically acceptable salts thereof. The definition of R is stated as in a specification. The invention also relates to a preparation method of the compounds and discloses a pharmaceutical composition which takes the compounds and the pharmaceutically acceptable salts as effective active components and the application in antifungal medicine.

Description

The verivate of pyrazole, Preparation Method And The Use

Technical field

The invention belongs to medical technical field, or rather, relate to one type of compound, its preparation method, compsn and application with anti-mycotic activity.

Background technology

Fungi infestation is invaded position difference according to it can be divided into two types in superficial part and deep.Mycotic infection of superficial part is a kind of common disease, frequently-occurring disease, is more common in skin and first portion; What deep fungal was more common is Candida albicans and Cryptococcus neoformans, though the low harm of sickness rate is bigger, but infects threat to life when serious.Particularly in recent decades; Along with being widely used of broad-spectrum antibiotics, cortin, antitumor drug and immunosuppressor; Extensively carrying out of radiotherapy and organ transplantation, generally the carrying out of conduit and intubate, and especially AIDS patient's increase rapidly of immunodeficient patient; Destroyed the symbiotic relationship of normal flora; Human body is reduced the resistibility of fungi, thus cause fungi infestation particularly deep fungal infection rise significantly, and become one of major disease main causes of death such as AIDS and tumour.

In the existing antifungal drug, the nitrogen azole drug is to have a type of better prospect, has obtained clinically using widely.The nitrogen azole drug is especially comparatively desirable to the curative effect of deep fungal infection, so be the focus in the antifungal drug research always.They can suppress fungal cell's cytochrome p 450 3A and rely on C14-α-demethylase (ERG11), and this enzyme is the key enzyme that changes into lanosterol ergosterol.Its mechanism of action is a blocking-up substrate hydroxylation reaction, and the interior methylated sterol of lanosterol 14-of fungus body is accumulated in a large number, and ergosterol is synthetic to be lacked, and causes many enzyme activity changes on membrane permeability and the film, thereby suppresses fungi growth.

Treat at present the choice drug of deep fungal infection clinically; Mainly contain with KETOKONAZOL (Ketoconazole), miconazole (Miconazole), tioconazole (Tioconazole) and be the glyoxaline compound of representative and be the Research of Triazole Antifungal Agents of representative with fluconazole (Fluconazole), itraconazole (Itraconazole) and Vorionazole (Voriconazole).But along with long-term widespread use, engender Resistant strain, and the trend of showed increased is arranged existing nitrogen azole drug.The existing Candida albicans B41628 that reports is to existing multiple medicine resistance, and the resistance problem makes the mycosis control face more stern challenge.In clinical large-scale experiment for many years, find simultaneously existing medicine, have to a certain degree toxic side effect such as hepatic disorder, gi tract infringement, fash etc. mostly, cause its clinical application to be restricted.

The limitation that exists to existing medicine is like narrow antimicrobial spectrum, resistance is serious, bioavailability is low, stronger problems such as toxic side effect are arranged.Except the preparation of optimizing the existing medicine of improvement, make the research of novel anti fungi-medicine become one of hot fields of global new drug development.In addition, the interaction between medicine makes the deep fungal infection treatment become complicated more, therefore, and the clinical novel deep antifungal drug that presses for high-efficiency low-toxicity, has a broad antifungal spectrum, highly selective.

Summary of the invention

One object of the present invention is, in order to overcome the deficiency of existing nitrogen azole compounds, develops the novel anti fungi-medicine, discloses the verivate and the pharmaceutical salts thereof of pyrazole.

Another object of the present invention is, discloses the preparation method of compound of Formula I and pharmaceutical salts thereof.

A further object of the present invention is, discloses the pharmaceutical composition that compound of Formula I and pharmaceutical salts thereof are main active ingredient.

A further object of the invention is, discloses compound of Formula I and pharmaceutical salts thereof the purposes as the antifungal drug aspect.

Combine goal of the invention that content of the present invention is specifically described at present.

The compound or the salt of general formula I structure involved in the present invention are following:

Wherein:

R is hydrogen or

Compound among the present invention with formula I structure, optional from following compound:

I-1 5-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine also;

I-2 (5-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine-2-yl also) acetic ester.

Compound with formula I structure of the present invention, its salt means: the salt that The compounds of this invention is become with mineral acid, organic acid.Wherein preferred especially salt is: hydrochloride, hydrobromate, hydriodate, vitriol, hydrosulfate, phosphoric acid salt, hydrophosphate, acetate, propionic salt, butyrates, lactic acid salt, mesylate, pharmacy acceptable salts such as tosilate, PHENRAMINE MALEATE, benzoate, SUMATRIPTAN SUCCINATE, tartrate, Citrate trianion, fumarate, taurate, gluconate.

The synthetic route map Succinct representation of the compound of formula I structure is following:

The hydrochloride of thienopyridine and 5-brooethyl-1; 3-dimethyl--1H-pyrazoles is in methylene dichloride, trichloromethane, toluene, ethanol, acetone equal solvent; In the presence of acid binding agents such as triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide ,-10 ℃～85 ℃ reactions make I-1.

The hydrochloride of Thienopyridinone and 5-brooethyl-1; 3-dimethyl--1H-pyrazoles is in methylene dichloride, trichloromethane, acetonitrile or toluene equal solvent; In the presence of acid binding agents such as triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide ,-10 ℃～105 ℃ reactions make key intermediate 4Midbody 4Again with diacetyl oxide, acetate, Acetyl Chloride 98Min. or acetyl bromide etc.; In methylene dichloride, trichloromethane or toluene; In the presence of acid binding agents such as triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide ,-30 ℃～50 ℃ reactions make compound I-2.

The reaction products therefrom is dissolved in a kind of among ether, DMF, acetone, methyl alcohol, ethanol, ETHYLE ACETATE or the DMSO, and dropping inorganic acid or organic acid solution are processed pharmacy acceptable salt.

Specifically be that all cpds is dissolved in a kind of among ether, DMF, acetone, methyl alcohol, ethanol, ETHYLE ACETATE or the DMSO, the dripping hydrochloric acid diethyl ether solution is processed hydrochloride to pH=2 under ice-water bath; Or all cpds is dissolved in a kind of among ether, DMF, acetone, methyl alcohol, ethanol, ETHYLE ACETATE or the DMSO, and adding and wait a mole glucono-, heated and stirred gets its gluconate; Or all cpds is dissolved in a kind of among ether, DMF, acetone, methyl alcohol, ethanol, ETHYLE ACETATE or the DMSO, drips the vitriol oil down to pH=3, process vitriol in ice-water bath, or the like.

This compounds is effective for the human fungal infectious disease of treatment.Although compound of the present invention can be without the direct administration of any preparation, described all cpds preferably uses with the form of pharmaceutical prepn, and route of administration can be non-enteron aisle approach (like vein, muscle administration) and oral administration.

The preparation of pharmaceutical compositions method of The compounds of this invention is following: use standard and conventional technology; Acceptable solid or liquid vehicle are combined, and make it at random with technology of pharmaceutics on acceptable auxiliary and vehicle combine to be prepared into particulate or microballoon.Solid dosage comprises tablet, discrete particles, capsule, slow releasing tablet, sustained release pellet or the like.Solid carrier can be at least a material, and it can serve as thinner, flavouring agent, solubilizing agent, lubricant, suspension agent, tackiness agent, disintegrating agent and coating agent.Inert solid carrier comprises trimagnesium phosphate, Magnesium Stearate, smoothers sugar, lactose, pectin, Ucar 35, Polysorbate 80, dextrin, starch, gelatin, cellulose substances for example methylcellulose gum, Microcrystalline Cellulose, low melt point paraffin, polyoxyethylene glycol, N.F,USP MANNITOL, theobroma oil etc.Liquid dosage form comprises solvent, suspension-s for example injection, pulvis or the like.

The amount of the active ingredient that contains in pharmaceutical composition and the unit dosage form (The compounds of this invention) can specifically be applied according to patient's the state of an illness, the situation of diagnosis, and the amount of used compound or concentration are regulated in the scope of a broad.Usually, the scope of active compound amount is 0.05%～90% (weight) of compsn, and another preferred range is 0.05%-70%.

Compound or its pharmacy acceptable salt with formula I structure of the present invention has the obvious suppression effect to fungi.Adopt standard micro-dilution method (NCCLSM-27A) to measure its minimum inhibitory concentration below, further specify the external bacteriostatic activity of The compounds of this invention clinical several frequently seen fungi.Concentration (the MIC that suppresses selected fungi 90% growth with target compound ₉₀) as judging terminal point.

Test strain: 5 kinds of deep fungals: Candida albicans (Candida albicans; C.alb.) ATCC 76615, Cryptococcus neoformans (Cryptococcus neoformans; C.neo.) ATCC 32609, Oidium tropicale (Candida tropicalis; C.tro.) ATCC 12034, Candida parapsilosis (Candida paropsilosis, C.par.) ATCC 22019, smoke aspergillus fumigatus (Aspergillus fumigatus, A.fum.).3 kinds of superficial fungis: trichophyton (Trichophyton rubrum, T.rub.), sporotrichum schenckii (Sporothrixschenckii, S.sch.) and fonsecaea pedrosoi (Fonsecaea pedrosoi, F.ped.).Above bacterial strain provides by our unit pharmacological experiment chamber.

Test drug: The compounds of this invention I-1, I-2; Select for use amphotericin B (AmB) (magnificent medicine group) and KETOKONAZOL (KCZ) (mesophytization Manufacturing Co., Ltd is closed in Wuhan) as positive control drug; DMSO 99.8MIN. (DMSO), AR level (the triumphant letter chemical industry in Tianjin ltd).ELIASA, BIO-RAD680.

TP: with RPMI-1640 (Sigma) is nutrient solution, and it is 1 * 10 that experimental strain is made into concentration ⁴～1 * 10 ⁵The suspension of/ml.Compound with sterile purified water and DMSO dissolving, is processed medicine storage liquid (6400 μ g/ml), and-20 ℃ of preservations are for use.Be diluted to 640 μ g/ml with RPMI-1640 during use, establish solvent control and blank simultaneously.Get the flat microtest plate in aseptic 96 holes; No. 1 hole adds RPMI-1640 100 μ L and makes blank, and No. 2 the hole adds bacteria suspension 180 μ L and soup 20 μ L, and the 3-12 hole respectively adds bacteria suspension 100 μ L; The 10 grades of doubling dilutions in 2-11 hole, 100 μ L of sucking-off abandon from No. 11 holes at last.Each hole drug level is respectively 64,32,16,8,4,2,1,0.5,0.25,0.125 μ gmL ^-1No. 12 the hole does not add soup, makes the medicine negative control.Candida is observed the growing state of 3d in 35 ℃ of constant temperature culture 24h.Cryptococcus neoformans is measured the result after cultivating 72h, and 26 ℃ of constant temperature culture of superficial mycosis are observed the growing state in 1 week.Analyze with ELIASA, compare with the medicine negative control hole, suppressing pairing lowest drug concentration with 90% is its minimal inhibitory concentration value (MIC ₉₀).

Compound extracorporeal antifungal activity experiment (MIC ₉₀, μ gmL ^-1)

Visible through external antibacterial test experiments result, compound I-1 all has inhibition to a certain degree active with I-2 to 8 kinds of pathomycetes, the first-selection that has even surpass the clinical treatment fungi but the bigger medicine amphotericin B of toxicity.Part of compounds to clinically the most common with produce drug-fast monilial infection on a large scale and have stronger external activity.This type new compound has wide spectrum, highly active advantage, has further further investigation and is worth.

Embodiment

Below in conjunction with embodiment the present invention is done further explanation, it is indicative that embodiment is merely, and means that never it limits scope of the present invention by any way.Described compound is through performance liquid chromatography (HPLC), and thin-layer chromatography (TLC) detects.Can adopt subsequently such as ir spectra (IR), nuclear magnetic resonance spectrum ( ¹H NMR, ¹³C NMR), mass spectrum (MS) etc. is further proved conclusively its structure.

Embodiment 1:

5-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine (compound I-1) also

In the reaction flask that stirring, condensing surface, TM are housed, add 18.9g (0.1mol) 5-brooethyl-1,3-dimethyl--1H-pyrazoles adds the 50mL absolute ethyl alcohol again, stirs to add Anhydrous potassium carbonate 27.6g (0.2mol) down.Add 17.6g (0.1mol) 4,5,6 then, the 7-THTP is the hydrochloride of [3,2-c] pyridine also, 25 ℃ of about 5h of insulation reaction (the flaggy demonstration reacts completely).Rf=0.44 [developping agent: v (sherwood oil): v (ETHYLE ACETATE)=1: 1].Stopped reaction filters, and filtrate decompression is steamed ethanol to the greatest extent, and post separates, and gets faint yellow oily product (HPLC:99.1%). ¹H?NMR(DMSO-d6，400MHz)δ：2.147(s，3H)，3.079～3.120(d，1H)，3.247～3.277(d，1H)，3.411(s，1H)，3.698(s，1H)，3.851(s，3H)，4.205～4.307(t，2H)，4.502～4.527(d，2H)，6.413(s，1H)，6.915～6.928(d，1H)，7.443～7.456(d，1H)。MS(m-1)/z：246.0。

Embodiment 2:

Midbody 4Preparation

In the reaction flask that stirring, condensing surface, TM are housed, add 14.5g (0.1mol) 5-brooethyl-1,3-dimethyl--1H-pyrazoles adds the 35mL methylene dichloride again, stirs to add triethylamine 20.2g (0.2mol) down.Add 19.2g (0.1mol) 5,6,7 then, the 7a-THTP is the hydrochloride of [3,2-c] pyridines-2 (4H)-ketone also, the about 3h of back flow reaction (the flaggy demonstration reacts completely).Rf=0.35 [developping agent: v (sherwood oil): v (ETHYLE ACETATE)=1: 2].Stopped reaction with 3 * 40mL water washing reaction solution, is obtained dichloromethane layer, uses the SODIUM SULPHATE ANHYDROUS 99PCT thorough drying, and methylene dichloride is to the greatest extent steamed in filtration, decompression, and post separates, and gets yellow oil product (HPLC:97.5%).

Embodiment 3:

(5-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine-2-yl also) acetic ester (compound I-2)

In the reaction flask that stirring, condensing surface, TM are housed, add 2.6g midbody 4, it is disperseed, stir adding sodium hydroxide 1.4g down with the 10mL trichloromethane.Reaction system is cooled to-20 ℃, the 1.1g diacetyl oxide is added reaction system in batches.Add, under room temperature, continue stirring reaction 1.5h (the flaggy demonstration reacts completely).With 3 * 15mL water washing reaction solution, obtain the trichloromethane layer, use the SODIUM SULPHATE ANHYDROUS 99PCT thorough drying, to filter, trichloromethane is to the greatest extent steamed in decompression, and post separates, and promptly gets light yellow oily product (HPLC:99.0%).Rf=0.67 [single-point, developping agent: v (sherwood oil): v (ETHYLE ACETATE)=1: 1]. ¹H?NMR(DMSO-d6，400MHz)δ：2.146(s，3H)，2.280(s，3H)，2.963～3.003(d，1H)，3.178～3.192(d，1H)，3.400～3.434(t，1H)，3.683(s，1H)，3.861(s，3H)，4.149(s，2H)，4.503～4.521(s，2H)，6.437(s，1H)，6.596(s，1H)。MS(m-1)/z：304.0。

Embodiment 4:

Compound I-1 one-tenth hydrochloride: get I-1 oily product 2.0g, be dissolved in the 10mL anhydrous diethyl ether.Ice-water bath is cooled to 0 ℃, drip 20.5% hydrochloric acid diethyl ether solution to pH be 2, continue at stir about 1h under the ice-water bath.Filter, get white solid.

Embodiment 5:

Compound I-2 one-tenth gluconate: get I-2 oily product 2.5g, be dissolved in the 15mL absolute ethyl alcohol.Be heated to the back adding that refluxes and wait a mole glucono-, continue at the about 3h of stirring reaction down that refluxes.Reaction finishes, and under room temperature, leaves standstill 24h.Filter, get white solid.

Embodiment 6:

Compound I-2 one-tenth vitriol: get I-2 oily product 2.3g, be dissolved in the 15mL anhydrous methanol.Ice-water bath is cooled to 5 ℃, drip concentrated sulfuric acid solution to pH be 3, continue at stir about 2h under the ice-water bath.Filter, get white solid.

For the pharmaceutical composition of pyrazole derivatives of the present invention is described more fully, following FORMULATION EXAMPLE is provided below, said embodiment only is used for explanation, rather than is used to limit scope of the present invention.Said preparation can use any active compound and the salt thereof in the The compounds of this invention, preferably uses embodiment 1,3,4,5, the compound described in 6.

Embodiment 7:

Prepare hard gelatin capsule with following compositions:

Consumption/capsule

Compound I-1 75mg

Pregelatinized Starch 100mg

Prist 4mg

Sodium starch glycolate 10mg

Magnesium Stearate 20mg

10% polyvidone ethanolic soln is an amount of

Preparation technology: supplementary material is dry in advance, and it is subsequent use to cross 100 mesh sieves.Press recipe quantity with the mentioned component mixing, cross 60 mesh sieves three times, add an amount of 10% polyvidone ethanol (95%) solution system softwood, cross 18 mesh sieves and granulate, 40 ℃ of dryings are crossed the whole grain of 16 mesh sieves, are packed in the hard gelatin capsule.

Embodiment 8:

Prepare tablet with following compositions:

Consumption/sheet

Compound I-2 75mg

Starch 45mg

Microcrystalline Cellulose 40mg

CMS sodium salt 4.5mg

Magnesium Stearate 1mg

Talcum powder 1mg

Prist 3mg

Preparation technology: supplementary material is dry in advance, and it is subsequent use to cross 100 mesh sieves.Earlier with the abundant mixing of the auxiliary material of recipe quantity.Bulk drug is added in the auxiliary material to increase progressively dilution method, and each abundant mixing of added-time 2-3 time guarantees medicine and the abundant mixing of auxiliary material; Cross 20 mesh sieves, dry 2h in 55 ℃ of ventilated drying ovens, dried particle cross the arrangement of 16 mesh sieves; Measure midbody content, mix compressing tablet on tabletting machine.

Embodiment 9:

The preparation of injection liquid:

The hydrochloride 45mg of compound I-1

Ucar 35 100mg

Polysorbate 80 is an amount of

Zero(ppm) water 300mL

Preparing method: get activeconstituents and join in the water for injection that dissolves sorbyl alcohol and Ucar 35, add medicinal basic adjusting pH value to 4～8 and make its dissolving.Add gac, whip attachment 30min, carbon removal, smart filter, embedding, sterilization.

Embodiment 10:

The preparation of injection lyophilized powder:

The gluconate 50mg of compound I-2

Medicinal basic 0.1-7.0%

N.F,USP MANNITOL 55-80%

Preparing method: get activeconstituents and add water for injection, regulate pH value to 4～8 with medicinal basic and make its dissolving.Add N.F,USP MANNITOL again, carry out autoclaving by the requirement of injection, add gac, adopt filtering with microporous membrane, filtrating is carried out packing, adopts freeze-drying, makes loose block, seals, and promptly gets.

Claims (7)

1. the compound and the pharmaceutical salts thereof that have formula I structure:

Wherein:

R is hydrogen or

2. compound and the pharmaceutical salts thereof with formula I structure as claimed in claim 1 is selected from following compound:

I-15-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine also;

I-2 (5-((1,3-dimethyl--1H-pyrazoles-5-yl) methyl)-4,5,6, the 7-THTP is [3,2-c] pyridine-2-yl also) acetic ester.

3. have the compound and the salt thereof of formula I structure, its salt refers to: the salt that formula I compound is become with mineral acid, organic acid,

Wherein:

R is hydrogen or

4. formula I compound and the salt thereof described in claim 3; Wherein preferred especially salt is: hydrochloride, hydrobromate, hydriodate, vitriol, hydrosulfate, phosphoric acid salt, hydrophosphate, acetate, propionic salt, butyrates, lactic acid salt, mesylate, tosilate, PHENRAMINE MALEATE, benzoate, SUMATRIPTAN SUCCINATE, tartrate, Citrate trianion, fumarate, taurate, gluconate.

5. the preparation method of claim 1 Chinese style I compound is characterized in that: 4,5; 6, the 7-THTP is the hydrochloride and the 5-brooethyl-1 of [3,2-c] pyridine also; 3-dimethyl--1H-pyrazoles is in methylene dichloride, trichloromethane, toluene, ethanol, acetone; In the presence of triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide ,-10 ℃～85 ℃ reactions make like compound I in the claim 2-1

5; 6,7,7a-THTP also [3; 2-c] hydrochloride and the 5-brooethyl-1 of pyridine-2 (4H)-ketone; 3-dimethyl--1H-pyrazoles is in methylene dichloride, trichloromethane, acetonitrile or toluene, and in the presence of triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide ,-10 ℃～105 ℃ reactions make key intermediate 4, midbody 4Again with diacetyl oxide, acetate, Acetyl Chloride 98Min. or acetyl bromide; In methylene dichloride, trichloromethane or toluene; In the presence of triethylamine, pyridine, salt of wormwood, yellow soda ash, sodium hydrogencarbonate, saleratus, sodium hydroxide or Pottasium Hydroxide;-30 ℃～50 ℃ reactions make like compound I in the claim 2-2

6. one kind has the anti-fungal activity medicaments compsn, and it comprises formula I compound as claimed in claim 1 and pharmaceutical salts and one or more pharmaceutical excipients of treating significant quantity.

Like each formula I compound and pharmaceutical salts thereof in the claim 1～2 in the application aspect the preparation antifungal drug.