Summary of the invention
The object of the present invention is to provide a kind of replacement benzothiazole compound and its esters.Another object of the present invention is to provide described replacement benzothiazole compound and the preparation method of its esters.The third object of the present invention is to provide described replacement benzothiazole compound and its esters preparing the application in antifungal drug, topoisomerase enzyme inhibitor, antitumor drug, antiviral, antihypertensive drug or antithrombotic reagent.
Technical scheme of the present invention is: in order to spread F TR13335 antimicrobial spectrum and improve its anti-mycotic activity; the naphthalene nucleus part mainly concentrating on FTR13335 carries out structure of modification; obtain the cis-N-{2-of a class [(pentamethylene base carbonyl) is amino] benzothiazol-6-yl }-3-[(replacement) is amino] cyclohexyl Carbox amide, can as the inhibitor of fungi N-mnyristoyl based transferase (NMT).
A first aspect of the present invention, be to provide a kind of replacement benzothiazole compound, i.e. cis-N-{2-[(pentamethylene base carbonyl) is amino] benzothiazol-6-yl }-3-[(replacement) is amino] cyclohexyl Carbox amide, and pharmacy acceptable salt, its general structure is as shown in formula I:
In formula I, R represents replacement naphthalene nucleus or substituted aroma heterocycle,
A) the various substituting group positions replaced in naphthalene nucleus or substituted aroma heterocycle are other any positions being positioned at removing link position, can be monosubstituted, also can be polysubstituted, substituting group refers to: hydrogen, hydroxyl, methyl, ethyl, methoxyl group, oxyethyl group, nitro, cyano group, trifluoromethyl or halogen, and described halogen comprises F, Cl, Br, I;
B) substituted aroma heterocycle represents independently: quinoline, tetrahydroquinoline, isoquinoline 99.9, tetrahydroisoquinoline, pyridine, thiophene, thiazole, pyrazoles, furans, oxazole, isoxazole, pyridazine or pyrazine etc.
Formula I compound, R group preferably from:
(1) R group is the bromo-2-naphthalene of 1-;
(2) R group is 6-methoxyl group-2-naphthalene;
(3) R group is 1-naphthalene;
(4) R group is 2-methoxyl group-1-naphthalene;
(5) R group is 4-methoxyl group-1-naphthalene;
(6) R group is 2-oxyethyl group-1-naphthalene;
(7) R group is 2-quinoline;
(8) R group is 3-quinoline;
(9) R group is the chloro-3-quinoline of 2-;
(10) R group is 1,2,3,4-tetrahydroquinoline-3-base;
(11) R group is 4-quinoline;
(12) R group is 6-quinoline;
(13) R group is 1,2,3,4-tetrahydroquinoline-6-base;
(14) R group is 8-quinoline;
Part preferred compound is:
Cis-N-{2-[(cyclopentylcarbonyl) is amino] benzothiazol-6-yl }-3-[(3-quinolylmethyl) is amino] cyclohexyl methane amide (8),
Cis-N-{2-[(cyclopentylcarbonyl) is amino] benzothiazol-6-yl }-3-[(6-quinolylmethyl) is amino] cyclohexyl methane amide (12), or
Cis-N-{2-[(cyclopentylcarbonyl) is amino] benzothiazol-6-yl }-3-[(1,2,3,4-tetrahydroquinoline-6-base) methylamino-] cyclohexyl methane amide (13).
Above-mentioned benzothiazole compound conventionally can prepare the form of a pharmaceutical salts.Comprise its organic acid salt and inorganic acid salt: mineral acid includes, but is not limited to hydrochloric acid, sulfuric acid, phosphoric acid, bisphosphate, Hydrogen bromide, nitric acid etc., organic acid includes, but is not limited to acetic acid, toxilic acid, fumaric acid, tartrate, succsinic acid, lactic acid, tosic acid, Whitfield's ointment, oxalic acid etc.
A second aspect of the present invention, is to provide a kind of preparation method replacing benzothiazole compound and pharmacy acceptable salt thereof.
The building-up reactions flow process of the compounds of this invention is as follows:
Concrete steps are:
(A) N-(6-nitrobenzene thiazole-2-base) ring penta methane amide (II) is prepared
2-amino-6-nitrobenzothiazole (I) and cyclopentane-carboxylic acid are in methylene dichloride, through 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and DMAP condensation, stirring at room temperature reacts 16 hours, generates N-(6-nitrobenzene thiazole-2-base) ring penta methane amide (II);
(B) N-(6-aminobenzothiazole-2-base) ring penta methane amide (III) is prepared
Compound N-(6-nitrobenzene thiazole-2-base) ring penta methane amide (II) and tin protochloride are in ethanol, and heating reflux reaction 2 hours, obtains N-(6-aminobenzothiazole-2-base) ring penta methane amide (III);
(C) the cis-3-of the tertiary butyl-N-[(2-[(cyclopentylcarbonyl) is amino] benzothiazole-6-is amino) carbonyl] cyclohexylcarbamate (IV) is prepared
N-(6-aminobenzothiazole-2-base) ring penta methane amide (III) and cis-3-(tertiary fourth oxygen carboxylic is amino) hexahydrobenzoic acid are in methylene dichloride, add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and I-hydroxybenzotriazole, stirring at room temperature reacts 18 hours, obtains the cis-3-of the tertiary butyl-N-[(2-[(cyclopentylcarbonyl) is amino] benzothiazole-6-is amino) carbonyl] cyclohexylcarbamate (IV);
(D) cis-3-amino-N-[2-(cyclopentylcarbonyl) is amino] benzothiazol-6-yl-cyclohexyl methane amide (V) is prepared
The cis-3-of the tertiary butyl-N-[(2-[(cyclopentylcarbonyl) is amino] benzothiazole-6-is amino) carbonyl] cyclohexylcarbamate (IV) is with methylene dichloride, add trifluoroacetic acid, stirring at room temperature reacts 1 hour, obtains cis-3-amino-N-[2-(cyclopentylcarbonyl) is amino] benzothiazol-6-yl-cyclohexyl methane amide (V);
(E) the cis-N-{2-of target compound [(cyclopentylcarbonyl) is amino] benzothiazol-6-yl is prepared }-3-[(replacement) is amino] cyclohexyl methane amide (VI)
The cis-3-amino of compound-N-[2-(cyclopentylcarbonyl) amino] benzothiazol-6-yl-cyclohexyl methane amide (V) and replacement naphthaldehyde or substd quinolines formaldehyde are in methyl alcohol, and mixed solution stirring at room temperature reacts 2 hours; Add sodium cyanoborohydride and 3-5 drips acetic acid, continue stirring reaction 10 hours, generate and prepare the cis-N-{2-of target compound [(cyclopentylcarbonyl) is amino] benzothiazol-6-yl-3-[(replacement) is amino] cyclohexyl methane amide (VI).
A third aspect of the present invention, is to provide described replacement benzothiazole compound and pharmacy acceptable salt is preparing the application in antifungal drug, topoisomerase enzyme inhibitor, antitumor drug, antiviral, antihypertensive drug or antithrombotic reagent.
Compound of the present invention is through antibacterial experiment in vitro, prove that majority of compounds has good anti-mycotic activity and wider antimicrobial spectrum, especially compound broad spectrum is greatly improved, and illustrates that this compounds can be used for preparing the medicine for the treatment of anti-fungal infection.
Compound of the present invention is that further investigation and exploitation new texture type antifungal drug open new approach.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described.Should be understood that following examples only for illustration of the present invention but not for limiting scope of the present invention.
The preparation of embodiment 1:N-(6-nitrobenzene thiazole-2-base) ring penta methane amide (II)
Get 2-amino-6-nitrobenzothiazole (4.00g, 0.02mol, 1equiv) be dissolved in 50mL methylene dichloride with cyclopentane-carboxylic acid (2.28g, 0.02mol, 1equiv), add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (5.75g, 0.03mol, 1.5equiv) and DMAP (0.40g, 2mmol, 0.1equiv), stirring at room temperature reaction 16h.After reaction terminates, reaction solution 5% salt pickling (3 × 30mL), 5% sodium bicarbonate is washed (3 × 30mL), and saturated nacl aqueous solution is washed (3 × 30mL), uses anhydrous Na
2sO
4dry. filter, evaporated under reduced pressure solvent, residue by silicagel column chromatography purification (developping agent: hexane:EtOAc=10:1, v/v), obtains faint yellow solid 5.66g, yield 97.3%.
1H-NMR(300MHz,CDCl
3):9.15(brs,1H),8.76(d,1H,J=2.4Hz),8.34(dd,1H,J=9.0,2.4Hz),7.82(d,1H,J=9.0Hz),2.80-2.96(m,1H),1.90-2.11(m,2H),1.54-1.90(m,6H).ESI-MS(m/z):290[M-1].
The preparation of embodiment 2:N-(6-aminobenzothiazole-2-base) ring penta methane amide (III)
Get compound ii (1.00g, 3.4mmol, 1equiv) and tin protochloride (2.30g, 10.2mmol, 3equiv) is dissolved in 50mL ethanol, heating reflux reaction 2h.After reaction terminates, evaporate to dryness reaction solution, adds 5% sodium hydroxide solution 50mL and 100mL ethyl acetate fully mixes vibration, and aqueous layer with ethyl acetate (2 × 100mL) extracts.Organic phase merges, and washes (3 × 30mL), anhydrous Na with saturated nacl aqueous solution
2sO
4dry.Filter, evaporated under reduced pressure solvent, residue with ethyl acetate 20mL recrystallization, obtains white solid 0.45g, yield 50.6%.
1H-NMR(300MHz,DMSO-d
6):11.97(s,1H),7.37(d,1H,J=8.5Hz),6.97(d,1H,J=1.5),6.68(dd,1H,J=8.5,1.5Hz),5.14(s,2H),2.83-2.97(m,1H),1.78-1.95(m,2H),1.45-1.95(m,6H).ESI-MS(m/z):262[M+1].
Embodiment 3: the preparation of the cis-3-of the tertiary butyl-N-[(2-[(cyclopentylcarbonyl) is amino] benzothiazole-6-is amino) carbonyl] cyclohexylcarbamate (IV)
Get III (1.00g, 3.83mmol, 1equiv) be dissolved in 25mL methylene dichloride with cis-3-(tertiary fourth oxygen carboxylic is amino) hexahydrobenzoic acid (0.93g, 3.83mmol, 1equiv), add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (1.10g, 5.75mmol, 1.5equiv) and I-hydroxybenzotriazole (0.78g, 5.75mmol, 0.1equiv), stirring at room temperature reaction 18h.After reaction terminates, filter out white depositions, use 20mL washed with dichloromethane, obtain faint yellow solid 0.91g, yield 54.2%.1H-NMR(300MHz,DMSO-d6):12.45(s,1H),10.21(s,1H),8.50(d,1H,J=1.8Hz),7.83(d,1H,J=8.7Hz),7.68(dd,1H,J=8.7,1.8Hz),7.04(s,1H),3.07-3.22(m,1H),2.72-2.81(m,1H),2.55-2.66(m,1H),1.67-2.17(m,12H),1.57(s,9H),1.20-1.53(m,4H).ESI-MS(m/z):485[M-1].
Embodiment 4: the preparation of cis-3-amino-N-[2-(cyclopentylcarbonyl) is amino] benzothiazol-6-yl-cyclohexyl methane amide (V)
Get IV (0.35g, 0.72mmol, 1equiv) to be dissolved in 3mL methylene dichloride, add trifluoroacetic acid 3mL, stirring at room temperature reaction 1h, evaporated under reduced pressure solvent, adds 5% aqueous sodium carbonate 30mL and ethyl acetate 50mL, fully mixes vibration.Aqueous layer with ethyl acetate (2 × 20mL) extracts.Merge organic phase, wash (3 × 30mL) with saturated nacl aqueous solution, anhydrous Na
2sO
4dry.Filter, evaporated under reduced pressure solvent, obtains white solid 0.27g, yield 92.59%.
1H-NMR(300MHz,DMSO-d
6):10.08(s,1H),8.41(s,1H),7.73(d,1H,J=8.7Hz),7.62(d,1H,J=8.7Hz),4.58-5.52(brs,2H),2.99-3.16(m,1H),2.43-2.69(m,2H),1.60-2.16(m,12H),0.99-1.60(m,4H).ESI-MS(m/z):385[M-1].
Embodiment 5: cis-N-{2-[(cyclopentylcarbonyl) amino] benzothiazol-6-yl } preparation of-3-[(the bromo-2-menaphthyl of 1-) is amino] cyclohexyl methane amide (in table compound 1)
Getting compound is dissolved in methyl alcohol 10mL along the bromo-2-naphthaldehyde (0.091g, 0.39mmol, 1equiv) of V (0.15g, 0.39mmol, 1equiv) and 1-, mixed solution stirring at room temperature reaction 2h.Add sodium cyanoborohydride (0.025g, 0.39mmol, 1equiv) to reaction solution and 3-5 drips acetic acid, continue stirring reaction 10h.After reaction terminates, the direct silica gel mixed sample of reaction solution, combiflash companion purifying (developping agent: CH
2cl
2-MeOH=10:1, v/v), obtain faint yellow solid 0.081g, yield 33.9%.
Embodiment 6: cis-N-{2-[(cyclopentylcarbonyl) amino] benzothiazol-6-yl } preparation of-3-[(3-quinolylmethyl) is amino] cyclohexyl methane amide (in table compound 8)
Getting compound is dissolved in methyl alcohol 10mL along V (0.15g, 0.39mmol, 1equiv) and 3-quinoline aldehyde (0.061g, 0.39mmol, 1equiv), mixed solution stirring at room temperature reaction 2h.Add sodium cyanoborohydride (0.025g, 0.39mmol, 1equiv) to reaction solution and 3-5 drips acetic acid, continue stirring reaction 10h.After reaction terminates, the direct silica gel mixed sample of reaction solution, combiflash companion purifying (developping agent: CH
2cl
2-MeOH=10:1, v/v), obtain faint yellow solid 0.015g, yield 7.14%.
13C-NMR(75MHz,CD
3OD):177.51,175.78,159.40,152.23,148.37,146.62,138.70,136.00,133.85,131.65,129.97,129.28,129.28,129.07,128.68,121.64,120.42,113.54,57.33,47.74,46.10,45.30,34.05,31.34,31.34,31.21,30.07,27.05,27.05,24.67.ESI-MS(m/z):528[M+1].HR ESI-MS[M+1]:528.2443.
Embodiment 7: cis-N-{2-[(cyclopentylcarbonyl) amino] benzothiazol-6-yl } preparation of-3-[(6-quinolylmethyl) is amino] cyclohexyl methane amide (in table compound 12)
Getting compound is dissolved in methyl alcohol 10mL along V (0.15g, 0.39mmol, 1equiv) and 6-quinoline aldehyde (0.061g, 0.39mmol, 1equiv), mixed solution stirring at room temperature reaction 2h.Add sodium cyanoborohydride (0.025g, 0.39mmol, 1equiv) to reaction solution and 3-5 drips acetic acid, continue stirring reaction 10h.After reaction terminates, the direct silica gel mixed sample of reaction solution, combiflash companion purifying (developping agent: CH
2cl
2-MeOH=10:1, v/v), obtain faint yellow solid 0.022g, yield 10.59%.
13C-NMR(75MHz,CD
3OD):177.49,176.36,159.42,151.17,148.18,146.52,138.86,138.27,136.17,133.82,131.99,129.81,129.32,128.53,122.78,121.62,120.46,113.53,56.57,50.58,46.10,45.82,35.85,31.34,31.34,30.73,30.20,27.05,27.05,25.14.ESI-MS(m/z):528[M+1].HR ESI-MS[M+1]:528.2443.
Embodiment 8: cis-N-{2-[(cyclopentylcarbonyl) amino] benzothiazol-6-yl } preparation of-3-[(1,2,3,4-tetrahydroquinoline-6-base) methylamino-] cyclohexyl methane amide (in table compound 13)
Getting compound is dissolved in methyl alcohol 10mL along V (0.15g, 0.39mmol, 1equiv) and 6-quinoline aldehyde (0.061g, 0.39mmol, 1equiv), mixed solution stirring at room temperature reaction 2h.Add sodium cyanoborohydride (0.025g, 0.39mmol, 1equiv) to reaction solution and 3-5 drips acetic acid, continue stirring reaction 10h.After reaction terminates, the direct silica gel mixed sample of reaction solution, combiflash companion purifying (developping agent: CH
2cl
2-MeOH=10:1, v/v), obtain faint yellow solid 0.014g, yield 8.11%.Compound 13 prepares the by product obtained in compound 12 process.
In embodiment, agents useful for same is commercially available analytical pure.
In table 1, other compounds are with different replacement naphthalene nucleus or quinoline ring for synthesis material, as listed in table 1, repeat the step in embodiment 5, just can synthesize required replacement benzothiazole compound.
The chemical structure of table 1 preferred compound and
1h-NMR and MS data
Embodiment 9: cis-N-{2-[(pentamethylene base carbonyl) amino] benzothiazol-6-yl of the present invention's synthesis } antifungic action of-3-[(replacements) amino] cyclohexyl Carbox amide
(1) experimental technique: adopt conventional antibacterial experiment in vitro method (referring to: Antimicrob AgentsChemother1995,39 (5): 1169)
1. materials and methods
(1) experimental strain
This experiment has selected following 6 kinds of important human body cause illness's standard fungal bacterial strains as screening object, and fungal bacterial strain provides by Changhai hospital of The 2nd Army Medical College Mycology Lab (or purchased from institute of materia medica of the Chinese Academy of Sciences).
1) Candida albicans (Candida albicans, type strain SC5314);
2) Candida parapsilosis (Candida parapsilosis, clinical strain 0306392);
3) Candida glabrata (Candida glabrata, type strain 537)
4) Cryptococcus neoformans (Cryptococcus neoformans, type strain BLS108);
5) trichophyton (Trichophyton rubrum, clinical strain 0501124);
6) gypsum shape sporidiole bacteria (Microsporum gypseum, clinical strain Cmccfmza).
(2) test method
Bacteria suspension is prepared: a. Cryptococcus neoformans and candidiasis are cultivated 16 hours through YEPD liquid nutrient medium 35 DEG C, twice activation, with blood cell counting plate counting, with RPM1640 liquid nutrient medium adjustment concentration to 1 × 10
3~ 5 × 10
3individual/mL.B. thread fungus (trichophyton) cultivates (35 DEG C) one week and (28 DEG C) two weeks respectively through SDA inclined-plane, and twice activation, adds RPM1640 liquid nutrient medium and blow and beat with suction pipe, spore is made to be free in RPM1640 liquid, through four layers of filtered through gauze, counting, adjustment concentration to 1 × 10
3~ 5 × 10
3individual/mL.
Drug solution preparing: get testing compound of the present invention and be dissolved in methyl-sulphoxide, is made into the medicament storage liquid of 3.2mg/mL, tests front RPM1640 and is diluted to 320 μ g/mL.
Inoculation: 96 No. 1, orifice plate holes add RPM1640200 μ l and make blank, 3-12 hole respectively adds bacteria suspension 100 μ l, No. 2 holes add bacteria suspension 180 μ l and liquid 20 μ l, the drug level in 2-11 hole makes 9 grade of 2 doubling dilution, and each hole drug level is respectively 32,16,8,4,2,1,0.5,0.25,0.125 μ g/mL.No. 12 holes do not add liquid, make positive control.
Cultivate and detect: set Positive control wells optical density value (OD value) as 100%, with optical density value than Positive control wells lower than 80% lowest concentration of drug for minimal inhibitory concentration value (MIC).
(2) experimental result
Antibacterial experiment in vitro the results are shown in Table 2.
The antimycotic minimal inhibitory concentration value of table 2 part selected objective target Compound ira vitro (MIC
80, μ g/mL)
Above-mentioned experimental result shows, compared with positive control drug, in compound of the present invention, major part has good anti-mycotic activity and wider antimicrobial spectrum, and especially compound broad spectrum is greatly improved, and illustrates that this compounds can be used for preparing the medicine for the treatment of anti-fungal infection.
More than show and describe ultimate principle of the present invention, principal character and advantage of the present invention.The technician of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and specification sheets just illustrates principle of the present invention; the present invention also has various changes and modifications without departing from the spirit and scope of the present invention, and these changes and improvements all fall in the claimed scope of the invention.Application claims protection domain is defined by appending claims and equivalent thereof.