CN102464682A - Aminoglycoside compound, and synthesis method and application thereof - Google Patents

Abstract

The invention discloses an aminoglycoside compound, and a synthesis method and application thereof, and belongs to the field of carbohydrate drugs. The 4' site of aminoglycoside with a neamine unit is structurally modified to obtain a group of bioactive molecules tolerant to action of various aminoglycoside modifying enzymes, and the structural general formula of the aminoglycoside compound is shown in a formula (I). The in vitro antimicrobial tests show that the aminoglycoside compound shown in the formula (I) has good antibacterial activity and has good inhibitory effects especially for drug-resistant bacteria of expressive aminoglycoside-modifying enzymes. The drug-resistant problem of antibiotic is one of the great difficulty problems that need to be addressed urgently in the field of current medicine, and therefor, the application prospects of the invention are wide.

Description

Aminoglycoside compounds and compound method thereof and application

Technical field

The present invention relates to the structure of modification of aminoglycoside compounds; Relate in particular to one type 4 ' and modified and have the aminoglycoside compounds and the compound method thereof of anti-microbial activity; The invention still further relates to the pharmacological use of such aminoglycoside compounds, belong to the carbohydrate pharmaceutical field.

Background technology

The antibiotics resistance problem is the serious challenge that world medical circle faces at present.Research shows that this and long-term abuse of antibiotics have direct relation.Bacterial resistance has been quickened in antibiotic long-term use lack of standardization, and even to this day, the speed that bacterial resistance produces is far away faster than the speed of new drug development.Aminoglycoside antibiotics is one type of widely used clinically Broad spectrum antibiotics, and Gram-negative bacteria and positive bacteria are all had very strong sterilization effect, and clinical application history is above 60 years.Because applicating history is remote, add use lack of standardization, comparatively serious resistance today appearred in this type microbiotic.Present known antibiotic resistance mechanism of production mainly contains following several kinds: (1) bacterium is highly expressed the aminoglycoside modifying enzyme; (2) the effect target of antibiotic molecule changes; (3) bacterium reduces antibiotic picked-up or evolves out and initiatively pump the mechanism of antibiotic molecule.Wherein, the most general with the antibiotic modifying enzyme of bacterial expression.

Bacterium is highly expressed the modifying enzyme of Antibiotique composition through transgenation.These endonuclease capables carry out chemically modified to the crucial group of drug molecule, thereby cause the bonding force reduction of drug molecule and target molecule or lose the generation resistance.Common aminoglycoside antibiotics modifying enzyme has three major types at present, and they are respectively: aminoglycoside acetyltransferase (AAC), aminoglycoside adenylyl transferase (AAD), aminoglycoside phosphoryl transferase (APH).And below each big type diversified each kind in number different effects site is arranged.

To the resistance problem that the antibiotics modification enzyme causes, people generally adopt two kinds of methods to overcome.First kind is the suppressor factor of development enzyme.Such as the clavulanic acid of successful Application clinically, be exactly a kind of beta-lactamase inhibitor, with itself and β-Nei Xiananleikangshengsu coupling, can play the effect of saving antibiotic activity.But this application of policies difficulty just occurred in the situation of aminoglycoside antibiotics, mainly is because the resistance enzyme class of aminoglycoside antibiotics is numerous, and emerges in an endless stream, and is difficult to find a kind of general enzyme inhibitors.Although some investigators are arranged in the world also at the enzyme inhibitors of this type of exploitation medicine, the enzyme inhibitors that these New Developments are put on display often only works to the resistance enzyme of very low range.Therefore, such work has only theory significance and does not have actual value, does not have sophisticated medicine so far and comes into the market.The second method that the resistance problem people that cause to modifying enzyme adopt is exactly the microbiotic of the anti-enzyme of development.General method is according to the certain designed theory, and natural aminoglycoside is carried out structure of modification, obtains some semisynthetic compound molecules, observes their anti-microbial activity then.Use this strategy and succeed, existing now matured product is applied clinically, such as amikacin, and HBK, dibekacin, netilmicin, Etimicin or the like.This type aminoglycoside has structurally kept the basic framework of natural aminoglycoside molecule, but on the particular functional group or introduce new group, or some special groups of cancellation.These non-native molecules that newly obtain have kept anti-microbial activity on the one hand, can tolerate the effect of some modifying enzyme to a certain extent on the other hand and possess the activity to resistant organism.Although obtained these progress,, the appearance of these new drugs can not solve all resistance problems, and they often have only a limited range of application.In addition, rule of thumb,, often cross the Resistant strain that this medicine how long will not occur in case a new antibiotic entering is clinical; With present viewpoint, the war of people and pathogenic bacteria is endless seasaw battle.Therefore, the new aminoglycoside antibiotics of development is still current medical developer's important topic.

Summary of the invention

The present invention's technical problem at first to be solved is the deficiency that overcomes existing aminoglycoside medicaments kind; One type of new aminoglycoside compounds is provided; This aminoglycoside compounds has good antibacterial activity, particularly to expressing the also good fungistatic effect of resistant organism of aminoglycoside modifying enzyme.

The present invention's technical problem at first to be solved realizes through following technical scheme:

The aminoglycoside compounds of structure shown in the formula (I) or its pharmaceutically useful salt:

Formula (I)

Wherein, R ¹Be selected from Wasserstoffatoms, substituted alkoxyl group, substituted amino or substituted amido; Preferably, said substituted amino comprises dimethylamino or kharophen; Described substituted amido comprises the amino butyrylamino of 4-, piperidines-4-formamido-or cyclohexyl formamido-or 2-(2-toluyl amido) acetamido;

R ²Be selected from Wasserstoffatoms or hydroxyl;

R ³Be selected from amino or hydroxyl;

R ⁴Be selected from Wasserstoffatoms, β-D-ribofuranosyl, 3-oxygen-(2; 6-diamino--2; 6-dideoxy-β-L-Chinese mugwort Du pyrans glycosyl)-β-D-ribofuranosyl, 3-oxygen-[4-oxygen (α-D-mannopyranose base)-2,6-diamino--2 or 6-dideoxy-β-L-Chinese mugwort Du pyrans glycosyl]-β-D-ribofuranosyl;

R ⁵Be selected from Wasserstoffatoms, 3-amino-3-deoxidation-α-D-glucopyranosyl or 3-amino-3-deoxidation-4-methyl-α-D-xylopyranosyl;

R ⁶Be selected from Wasserstoffatoms, substituted acyl group, alkyl or substituted alkyl; Preferably, said substituted acyl group is 2-hydroxyl-4-amino-butyryl radicals; Said alkyl is preferably ethyl;

R ⁷Be selected from hydroxyl, amino or methylamino;

R ⁸Be selected from Wasserstoffatoms or methyl.

Certainly, can prepare the salt of The compounds of this invention acid addition, these salt are included in the present invention.

The acid salt of The compounds of this invention is preferably pharmaceutically acceptable, forms nontoxic salt with suitable acid (for example hydrochloric acid, acetic acid, sulfuric acid), and except pharmaceutically acceptable salt, other salt is also included among the present invention.

Another technical problem to be solved by this invention provides the method that a kind of preparation has the aminoglycoside compounds of structure shown in the above-mentioned formula (I).

Another technical problem to be solved by this invention realizes through following technical scheme:

The method of the aminoglycoside compounds of structure shown in a kind of above-mentioned formula of preparation (I) may further comprise the steps:

(1) with obtain after the aminoglycoside compound cracking shown in the formula II shown in the formula III removing ring I as glycosyl acceptor:

formula II

formula III

(2) 4 provisional protections shown in the preparation formula IV as glycosyl donor:

formula IV

(3) through glycosylation glycosyl acceptor formula III compound and glycosyl donor formula IV compound are carried out coupling, slough 4 ' protection base on the formula IV compound ring I after, further deriving connects target functional group, finally sloughs all protections, promptly gets.

Wherein, 3 ' and 4 ' hydroxyl of the ring I of the aminoglycoside compound molecule shown in step (1) the Chinese style II is exposed, and all the other all hydroxyls are with benzyl or the protection of other appropriate protection base, and are amino with nitrine form or other protection base protection; Then the gained midbody is used sodium periodate oxidation, again alkaline condition down mode such as degraded obtain formula III compound as glycosyl acceptor.

Formula IV compound described in the step (2) exists with sulphur glycosides or tribromo-acetyl polyurethane form; 4 hydroxyl or amino to be different from other hydroxyl or the amino protection base protection on glycosyl acceptor or the donor can optionally be removed after glycosylation is accomplished.

Glycosylation described in the step (3) can carry out under typical glycosylation condition, such as the sulphur glycosides glycosylation of NIS/TfOH mediation, and the perhaps acid catalyzed trichlorine polyurethane of Lewis glycosylation etc.;

Another technical problem to be solved by this invention provides a kind of pharmaceutical composition of treating infectation of bacteria; This pharmaceutical composition is cooperated with pharmaceutically acceptable carrier by the compound of structure shown in the formula (I) of significant quantity or its pharmaceutically useful salt and forms; Formula (I) compound that is about to pharmaceutically acceptable consumption is with after pharmaceutically acceptable carrier or thinner cooperate, and the formulation method conventional by this area is prepared into any one appropriate drug compsn with it.The aminoglycoside compounds of the present invention that is about to significant quantity is with after pharmaceutically acceptable carrier or thinner cooperate, and by the formulation method of this area routine it is prepared into any one appropriate drug compsn.Usually said composition is suitable for oral administration and drug administration by injection, also is fit to other medication.Said composition can be liquid preparation forms such as tablet, capsule, pulvis, granule, lozenge, suppository, or oral liquid.According to different medications, pharmaceutical composition of the present invention can contain 0.1%-99% weight, the aminoglycoside compounds of the present invention of preferred 10-60% weight.

The breviary term that arrives involved in the present invention

The TLC thin-layer chromatography

The DMF N

NIS iodo succimide

DIPEA N, the N-diisopropyl ethyl amine

TBTU benzotriazole tetrafluoride boron salt

Description of drawings

The structural formula of Fig. 1 aminoglycoside compounds of the present invention.

The synthetic route of Fig. 2 glycosyl acceptor of the present invention; A) 1) fluoroform sulfonyl azide, triethylamine, copper sulfate, acetonitrile, water; 2) aceticanhydride, pyridine; 3) sodium methylate, methyl alcohol; B) phenylbenzene tertiary butyl chloride silane, pyridine, 4-Dimethylamino pyridine, four step total recoverys 59%; C) 2,2-methoxy propane, methylene dichloride, camphorsulfonic acid, 52%; D) 1) tetrabutyl ammonium fluoride, THF; 2) sodium hydride, bromotoluene, N, dinethylformamide, two step total recoverys 82%; E) acetic acid, methyl alcohol, 100%; F) 1) sodium periodate, methyl alcohol; 2) n-Butyl Amine 99, methyl alcohol, two step total recoverys 88%.

The synthetic route of glycosyl donor among Fig. 3 the present invention; A) sodium hydride, bromotoluene, N, dinethylformamide, 92%; B) camphorsulfonic acid, methyl alcohol; C) 1) Tosyl chloride, pyridine; 2) sodium azide, N, dinethylformamide, three step total recoverys 81%; D) 1) Dess-Martin ' s oxygenant, methylene dichloride; 2) Peng Qinghuana, methyl alcohol, 95%; E) 1) Trifluoromethanesulfonic anhydride, pyridine, methylene dichloride; 2) ammonia, methyl alcohol; 3) 2,2,2-trichlorine ethoxy dicarbonyl chloride, sodium hydrogencarbonate, THF, three step yields 55%.

The synthetic route that the aminoglycoside molecule is rebuild among Fig. 4 the present invention; A) iodo succimide; Trifluoromethanesulfonic acid; 4

molecular sieve, methylene dichloride ,-70 ℃ to room temperature; 57%, α/β=1.74/1; B) sodium hydroxide, water, 1, the 4-dioxane refluxes 64%; C) sodium hydride, methyl iodide, N, dinethylformamide, 50%; D) aceticanhydride, pyridine, 80%; E) 1) 4-chloro-butyryl chloride, sodium hydrogencarbonate, THF; 2) sodium azide, N, dinethylformamide, 80%; F, g, h) benzotriazole tetrafluoride boron salt, carboxylic acid, diisopropyl ethyl amine, N, dinethylformamide.19,20,21 productive rates separately: 78%, 75%, 81%; I) 1) hydrogen sulfide, pyridine, triethylamine; 2) hydrogen, palladium carbon, methyl alcohol, water, hydrochloric acid, 22,23,24,25,26,27 productive rates separately: 95%, 87%, 66%, 61%, 64%, 60%.

Embodiment

Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment with form or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall in protection scope of the present invention the details of technical scheme of the present invention.

Embodiment 16 "-preparation (3) of silica-based-five azide kanendomycins of O-tert-butyl diphenyl

Under the ice bath 2.0 gram (4.14 mmole) kanendomycins are dissolved in 6 ml waters, add 33 milligrams of (0.21 mmole) copper sulfate, stirring and dissolving is complete.Add 4.18 gram (41.4 mmole) triethylamines then, stir the acetonitrile solution (1.5N, 31.5 mmoles) that adds 21 milliliters of fluoroform sulfonyl azides after 15 minutes.Ice bath stirred 2 hours down, removed ice bath, continued to stir 10 hours.Boil off solvent, crude product repeatedly with after the anhydrous methanol dissolving again evaporate to dryness to color turn to be yellow.Add diacetyl oxide (6 milliliters)/pyridine (9 milliliters) mixture stirred overnight then.With splashing in 200 ml waters under the reaction mixture stirring, separate out a large amount of depositions.Filter, filter cake is through being dissolved in chromatographic column separation on an amount of methylene dichloride after the dry air, with petrol ether/ethyl acetate system gradient (petrol ether/ethyl acetate V/V=6: 1-3: 1) wash-out.Collect the elutriant evaporate to dryness, products obtained therefrom and 60 ml methanol are mixed, and the methanol solution of sodium methylate that adds catalytic amount stirs.Question response finishes, and adds Zeo-karb and is neutralized to neutrality.Solvent evaporated gets full azide product 21.664 grams (2.71 mmole).Products therefrom is dissolved in 30 milliliters of pyridines, adds the 4-Dimethylamino pyridine of 1.5 gram (5.5 mmole) tert-butyl diphenyl chlorosilanes and catalytic amount, normal temperature stirred 48 hours down.Solvent evaporated, crude product are dissolved in chromatographic column on an amount of methylene dichloride, with petrol ether/ethyl acetate (petrol ether/ethyl acetate V/V=3: 1) wash-out.Collect the elutriant evaporate to dryness, get gel-like prod 2.08 grams (2.44 mmole), overall yield 58.5%.

¹H?NMR(500MHz，CDCl ₃)δ7.70-7.65(m，4H)，7.47-7.39(m，6H)，5.54(d，J＝3.5Hz，1H)，5.09(d，J＝3.5Hz，1H)，4.13-4.09(m，1H)，4.01-3.90(m，4H)，3.84(dd，J1＝4.0Hz，72＝11.0Hz，1H)，3.68-3.45(m，9H)，3.42-3.27(m，4H)，2.94(brs，3H)，2.42(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.77(brs，2H)，1.62(ddd，J1＝J2＝J3＝12.5Hz，1H)，1.08(s，9H). ¹³C?NMR(125MHz，CDCl ₃)δ135.60，135.55，132.74，132.64，130.00，129.95，127.88，127.83，99.40，98.02，84.06，80.12，75.29，72.10，72.01，71.37，71.22，71.18，70.03，66.30，63.80，63.12，58.90，58.78，51.17，31.54，26.82，19.21.HRESI-MS?Calcd?forC ₃₄H ₄₉N ₁₆O ₁₀Si([M+NH ₄] ⁺)：869.3581，Found：869.3584

Embodiment 23 ', 4 '-O-isopropylidene-6 "-preparation (4) of silica-based-five azide kanendomycins of O-tert-butyl diphenyl

0.705 gram (0.827 mmole) 3 (embodiment 1 preparation) is dissolved in 40 milliliters of methylene dichloride and 2, (V/V=1: 1), add the camphorsulfonic acid of catalytic amount, normal temperature stirred 24 hours down the mixture of 2-dimethoxy propane.Add three triethylamine cancellation reactions, solvent evaporated.Silica gel chromatographic column on the residue is with petrol ether/ethyl acetate gradient elution (petrol ether/ethyl acetate V/V=8: 1-3: 1).Get title product 0.236 gram.Collect polarity each isomer 0.222 gram less than title product, be dissolved in (15 milliliters/5 milliliters) in the methylene chloride-methanol mixture, add 20 milligrams of Zeo-karbs, normal temperature stirs down, and with TLC (thin-layer chromatography) detection reaction, reaction in about 3 hours finishes.With triethylamine cancellation reaction back solvent evaporated, residue separates with column chromatography, with petrol ether/ethyl acetate gradient elution (petrol ether/ethyl acetate V/V=6: 1-3: 1), get title product 0.145 gram.Merging gross weight is 0.381 gram (0.427 mmole), overall yield 52%.Product characters is the flakey white solid.The high polarity product that other collects changes the next round reaction over to.

¹H?NMR(500MHz，CDCl ₃)δ7.68-7.63(m，4H)，7.47-7.38(m，6H)，5.71(d，J＝4.0Hz，1H)，5.02(d，J＝3.0Hz，1H)，4.31-4.28(m，1H)，3.96-3.81(m，5H)，3.65-3.52(m，7H)，3.47-3.40(m，3H)，3.31-3.27(m，2H)，2.83(d，J＝9.5Hz，1H)，2.60(d，J＝3.0Hz，1H)，2.43(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.62(ddd，J1＝J2＝J3＝13.0Hz，1H)，1.47(s，3H)，1，46(s，3H)，1.06(s，9H). ¹³C?NMR(125MHz，CDCl ₃)δ135.60，135.55，132.67，132.60，130.00，129.94，127.88，127.83，111.76，100.06，98.06，85.37，79.15，75.68，75.20，74.98，72.37，72.07，71.43，69.92，66.34，63.81，61.56，59.05，58.60，51.68，31.48，26.81，26.74，26.43，19.19.Anal.Calcd?for?C ₃₇H ₄₉N ₁₅O ₁₀Si：C，49.82；H，5.54；N，23.55.Found：C，50.11；H，5.46；N，23.81.ESI-MS：Calcd.For?C ₃₇H ₅₃N ₁₆O ₁₀Si([M+NH ₄] ⁺)：909，Found?909.

Embodiment 33 ', 4 '-O-isopropylidene-5,2 ", 4 ", 6 "-preparation (5) of four-O-benzyl-five azide kanendomycin

0.632 gram (0.708 mmole) 4 (embodiment 2 preparations) is dissolved in 15 milliliters of THFs, adds tetrahydrofuran solution 0.748 gram (1.2N, density 0.88,0.85 mmole) of the tetrabutyl ammonium fluoride of 1M, normal temperature stirs down.After 30 minutes, the thin-layer chromatography detection reaction is accomplished.Solvent evaporated; Residue with chromatographic column on an amount of methylene dichloride after with petrol ether/ethyl acetate (V/V=3: 1) wash-out; Products therefrom desolventizes through fully steaming; Be dissolved in 10 milliliters of Ns, add sodium hydride and 698 milligrams of (4.1 mmole) bromobenzyls of 164 milligrams of (4.2 mmoles) 60%, normal-temperature reaction 4 hours.Processing reaction is separated through silica gel column chromatography, with petrol ether/ethyl acetate wash-out (petrol ether/ethyl acetate V/V=10: 1), get yellow gluey product 0.587 gram (0.579 mmole), productive rate 82%.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.41(m，2H)，7.39-7.36(m，2H)，7.33-7.20(m，11H)，7.14-7.06(m，3H)，6.98-6.96(m，2H)，5.63(d，J＝4.0Hz，1H)5.61(d，J＝4.0Hz，1H)，4.95(s，2H)，4.81(d，J＝12.0Hz，1H)，4.74(d，J＝12.5Hz，1H)，4.62(d，J＝11.0Hz，1H)，4.47(d，J＝12.5Hz，1H)，4.41-4.38(m，1H)，4.27-4.23(m，2H)，3.98(dd，J1＝9.0Hz，J2＝11.0Hz，1H)，3.82-3.78(m，2H)，3.72(t，J＝9.5Hz，1H)，3.65(t，J＝9.5Hz，1H)，3.58-3.52(m，3H)，3.47-3.23(m，7H)，3.16(dd，J1＝3.0Hz，J2＝11.0Hz，1H)，2.40(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.62(ddd，J1＝J2＝J3＝12.5Hz，1H)，1.45(s，3H)，1.43(s，3H). ¹³C?NMR(125MHz，CDCl ₃)δ137.85，137.59，137.33，137.20，128.49，128.30，128.23，128.15，128.10，128.06，127.75，127.72，127.47，127.21，126.16，111.82，97.74，95.83，83.23，77.81，77.48，77.19，75.89，75.06，74.99，74.77，74.46，73.48，73.08，72.21，70.10，67.74，65.31，61.22，60.18，59.37，51.70，31.97，26.70，26.38.Anal.Calcd?forC ₄₉H ₅₅N ₁₅O ₁₀：C，58.04；H，5.47；N，20.72.Found：C，57.83；H，5.55；N，20.46.ESI-MS：Calcd.For?C ₄₉H ₅₅N ₁₅O ₁₀K([M+K] ⁺)：1052，Found?1052.

Embodiment 45,2 ", 4 ", 6 "-preparation (6) of four-O-benzyl-five azide kanendomycin

2.92 gram (2.88 mmole) 5 (embodiment 3 preparations) is dissolved in 60 ml methanol, adds 1.5 milliliters of acetate, reflux 4 hours, and the TLC detection reaction finishes.Steaming desolventizes, and residue is eluent (petrol ether/ethyl acetate V/V=6: 1-3: 1) obtain yellow gluey product 2.80 grams (2.87 mmole), productive rate 100% through the column chromatography separation with the petrol ether/ethyl acetate mixed solution.

¹H?NMR(500MHz，CDCl ₃)δ7.42-7.36(m，4H)，7.34-7.18(m，11H)，7.12-7.07(m，3H)，6.96-6.94(m，2H)，5.59(d，J＝4.0Hz，1H)，5.47(d，J＝4.0Hz，1H)，4.98(d，J＝12.0Hz，1H)，4.92(d，J＝11.5Hz，1H)，4.80(d，J＝11.5Hz，1H)，4.74(d，J＝12.0Hz，1H)，4.60(d，J＝11.0Hz，1H)，4.46(d，J＝12.0Hz，1H)，4.26(d，J＝12.0Hz，1H)，4.22(d，J＝11.0Hz，1H)，4.19-4.15(m，1H)，3.91(dd，J1＝9.0Hz，J2＝10.0Hz，1H)，3.81-3.77(m.2H)，3.67-3.63(m，2H)，3.58-3.33(m，8H)，3.29(dd，J1＝1.5Hz，J2＝11.0Hz，1H)，3.13(dd，J1＝2.5Hz，J2＝11.0Hz，1H)，3.07(dd，J1＝4.0Hz，J2＝10.5Hz，1H)，2.82(brs，1H)，2.66(brs，1H)，2.37(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.63(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ137.79，137.56，137.30，128.53，128.33，128.26，128.20，128.13，128.10，127.77，127.52，127.17，125.92，97.48，95.90，82.81，77.78，77.49，75.92，74.86，74.47，73.51，73.18，71.97，71.20，71.07，70.09，67.74，65.28，62.86，60.07，59.18，51.21，31.75.HRESI-MS?Calcd?for?C ₄₆H ₅₂N ₁₅O ₁₀([M+H] ⁺)：974.4016；found：974.3997

Embodiment 51, the preparation (7) of 3-two nitrine-5-O-benzyl-6-O-(3-deoxidation-3-nitrine-2,4,6-three-O-benzyl-alpha-D-glucopyranosyl)-2-deoxystreptamine

4.60 gram (4.72 mmole) 6 (embodiment 4 preparations) is dissolved in 80 ml methanol, adds 2.02 gram (9.44 mmole) sodium periodates, normal temperature stirred 8 hours down.Reaction mixture is poured in 200 milliliters of salt solutions, with ethyl acetate extraction three times, each 40 milliliters.Combining extraction liquid evaporate to dryness, residue are dissolved in 200 ml methanol, stir to add 300 milligrams of n-Butyl Amine 99s, stirred overnight under the normal temperature down.Steaming desolventizes, and residue is through column chromatography for separation, and (petrol ether/ethyl acetate V/V=6: 1) wash-out gets white gluey product 3.18 grams (4.17 mmole), productive rate 88% with the petrol ether/ethyl acetate system.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.09(m，20H)，5.62(d，J＝3.5Hz，1H)，4.84-4.71(m，5H)，4.51(d，J＝12.0Hz，1H)，4.35(d，J＝11.0Hz，1H)，4.28(d，J＝12.5Hz，1H)，4.07-4.05(m，1H)，3.90(t，J＝10.0Hz，1H)，3.60-3.53(m，2H)，3.47-3.30(m，7H)，2.48(brs，1H)，2.30-2.25(m，1H)，1.53-1.45(m，1H). ¹³C?NMR(125MHz，CDCl ₃)δ137.87，137.66，137.39，137.28，128.49，128.32，128.24，128.18，128.09，127.97，127.75，127.70，95.61，82.45，77.34，76.91，76.86，76.32，76.14，74.80，73.41，72.93，69.88，67.74，65.48，60.72，59.84，31.76.Anal.Calcd?for?C ₄₀H ₄₃N ₉O ₇：C，63.06；H，5.69；N，16.55.Found：C，62.86；H，5.69；N，16.22.ESI-MS：Calcd.For?C ₄₀H ₄₃N ₉O ₇Na([M+Na] ⁺)：784，Found：784.

Embodiment 6 p-methylphenyls-2-deoxidation-2-nitrine-4, the preparation (8) of 6-O-benzal-1-sulphur-alpha-D-glucose glycosides

Make by literature method.(Chem.Eur.J.2007，13，529-540)。

¹H?NMR(500MHz，CDCl3)δ7.52-7.50(m，2H)，7.42-7.38(m，5H)，7.14(d，J＝8.0Hz，2H)，5.55(s，1H)，5.50(d，J＝5.5Hz，1H)，4.41(dt，J1＝5.0Hz，J2＝10.0Hz，1H)，4.23(dd，J1＝5.0Hz，J2＝10.0Hz，1H)，4.07(t，J＝9.5Hz，1H)，3.90(dd，J1＝6.0Hz，J2＝10.5Hz，1H)，3.75(t，J＝10.5Hz，1H)，3.57(t，J＝9.0Hz，1H)，2.79(br，1H)，2.34(s，3H)

Embodiment 7 p-methylphenyls-2-deoxidation-2-nitrine-3-O-benzyl-4, the preparation (9) of 6-O-benzal-1-sulphur-alpha-D-glucose glycosides

1.979 gram (4.95 mmole) 8 (embodiment 6 preparations) is dissolved among 25 milliliters of DMF, adds the sodium hydride of 0.396 gram (9.9 mmole) 60%, normal temperature stirred 20 minutes down, splashed into 1.693 gram bromobenzyls then.Continue to stir three hours, and reaction solution was poured in 150 mL of saline stirred, with ethyl acetate extraction three times, each 20 milliliters.Combining extraction liquid adds anhydrous sodium sulfate drying, filters the filtrating evaporate to dryness.Residue dissolves with an amount of methylene dichloride, adds proper silica gel absorption then, steaming vibrating dichloromethane.Change silica gel over to chromatographic column with petrol ether/ethyl acetate-dichloromethane mixture gradient elution (V/V/V=8: 1: 1-5: 1: 1), get colorless oil product 2.23 grams (4.55 mmole), productive rate 92%.

¹H?NMR(500MHz，CDCl ₃)δ7.52-7.49(m，2H)，7.42-7.28(m，10H)，7.13(d，J＝8.5Hz，2H)，5.59(s，1H)，5.49(d，J＝5.0Hz，1H)，4.97(d，J＝10.5Hz，1H)，4.83(d，J＝11.0Hz，1H)，4.44(dt，J1＝J2＝5.0Hz，J3＝10.0Hz，1H)，4.23(dd，J1＝5.0Hz，J2＝10.5Hz，1H)，3.99-3.93(m，2H)，3.78-3.73(m，2H)，2.34(s，3H). ¹³C?NMR(125MHz，CDCl ₃)δ138.34，137.65，137.11，133.10，129.97，129.08，128.42，128.30，128.23，127.94，125.99，101.47，88.14，82.74，77.81，75.17，68.59，63.69，63.57，21.14.Anal.Calcd?for?C ₂₇H ₂₇N ₃O ₄S：C，66.24；H，5.56；N，8.58.Found：C，66.08；H，5.552；N，8.50.ESI-MS：Calcd.ForC ₂₇H ₂₇N ₃O ₄SK([M+K] ⁺)：528，Found?528.

The preparation (11) of embodiment 8 p-methylphenyls-2-deoxidation-2-nitrine-3-O-benzyl-6-deoxidation-6-nitrine-1-sulphur-alpha-D-glucose glycosides

4.25 gram (8.68 mmole) 9 (embodiment 7 preparations) is dissolved in the mixture of 20 milliliters of methylene dichloride and 80 ml methanol, adds 100 milligrams of camphorsulfonic acids, reflux 6 hours.Solvent evaporated, residue are behind the chromatographic column purifying, and product is dissolved in 30 milliliters of pyridines, add 2.37 gram (12.4 mmole) Tosyl chlorides, and stirring at normal temperature is spent the night.Remove and to desolvate, residue is after chromatographic column is separated, and product is dissolved in 40 milliliters of DMF, adds 0.70 gram (10.8 mmole) sodium azide, 60 ℃ of heated and stirred 8 hours.Reaction solution is poured in 150 milliliters of salt solutions, with ethyl acetate extraction 3 times, each 40 milliliters.Collect the oil reservoir evaporate to dryness, residue is with ETHYLE ACETATE-sherwood oil system (petrol ether/ethyl acetate V/V=8: 1-4: 1) cross chromatographic column, get product 3.00 grams (7.03 mmole), productive rate 81%.

¹H?NMR(500MHz，CDCl ₃)δ7.42-7.38(m，6H)，7.37-7.33(m，1H)，7.14(d，J＝8.0Hz，2H)，5.52(d，J＝5.5Hz，1H)，5.01(d，J＝11.0Hz，1H)，4.71(d，J＝11.5Hz，1H)，4.33-4.29(m，1H)，3.89(dd，J1＝5.5Hz，J2＝10.5Hz，1H)，3.60(t，J＝8.5Hz，1H)，3.54(dt，J1＝J2＝3.5Hz，J3＝9.5Hz，1H)，3.49(dd，J1＝3.0Hz，J2＝13.0Hz，1H)，3.49(dd，J1＝5.5Hz，J2＝13.5Hz，1H)，2.34(s，3H)，2.15(d，J＝3.0Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ138.26，137.69，132.73，129.98，129.05，128.85，128.38，128.15，87.52，81.33，75.42，71.55，71.17，63.78，51.30，21.11.Anal.Calcd?for?C ₂₀H ₂₂N ₆O ₃S：C，56.32；H，5.20；N，19.70.Found：C，56.30；H，4.98；N，19.90.ESI-MS：Calcd.For?C ₂₀H ₂₆N ₇O ₃S([M+NH ₄] ⁺)：444，Found?444.

The preparation (12) of embodiment 9 p-methylphenyls-2-deoxidation-2-nitrine-3-O-benzyl-6-deoxidation-6-nitrine-1-sulphur-α-D-galactoside

1.20 gram (2.81 mmole) 11 (embodiment 8 preparations) is dissolved in 30 milliliters of anhydrous methylene chlorides, and adding 1.44 gram (3.39 mmole) Dess-Martin oxygenants (1,1,1-Triacetoxy-1,2-benziodoxol-3 (1H)-one) stirring at normal temperature 3 hours.In reaction mixture, add 30 milliliters of Sodium Pyrosulfites (containing 1.50 grams) and the sodium hydrogencarbonate mixed aqueous solution of (containing 3.0 grams) then, fully stirred 1 hour.Tell dichloromethane layer, water layer is used an amount of washed with dichloromethane, evaporate to dryness after the merging organic layer drying.Residue suspends with 20 ml methanol, and ice bath adds 120 milligrams of Peng Qinghuanas down and stirs.Along with the carrying out of reaction, reaction solution becomes clarification very soon.Treat that TLC detects raw material and all disappears, solvent evaporated, residue through chromatographic column with the petrol ether/ethyl acetate gradient elution (petrol ether/ethyl acetate V/V=8: 1-4: 1), configuration inversion product 0.99 gram (2.32 mmole), yield 95% (brsm).Reclaim initial pure 0.16 gram in addition.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.38(m，6H)，7.37-7.35(m，1H)，7.14(d，J＝8.0Hz，2H)，5.54(d，J＝5.5Hz，1H)，4.79(d，J＝11.5Hz，1H)，4.69(d，J＝11.5Hz，1H)，4.44-4.42(m，1H)，4.22(dd，J1＝5.0Hz，J2＝10.0Hz，1H)，3.99(t，J＝1.5Hz，1H)，3.76(dd，J1＝3.5Hz，J2＝10.5Hz，1H)，3.59(dd，J1＝7.5Hz，J2＝12.5Hz，1H)，3.36(dd，J1＝5.0Hz，J2＝13.0Hz，1H)，2.41(t，J＝1.5Hz，1H)，2.33(s，3H). ¹³C?NMR(125MHz，CDCl ₃)δ138.20，136.75，132.77，129.97，128.94，128.78，128.49，128.10，87.65，77.54，72.48，69.95，66.57，59.52，51.06，21.12.Anal.Calcd?for?C ₂₀H ₂₂N ₆O ₃S：C，56.32；H，5.20；N，19.70.Found：C，56.39；H，5.18；N，19.85.ESI-MS：Calcd.For?C ₂₀H ₂₆N ₇O ₃S([M+NH ₄] ⁺)：444，Found?444.

The preparation (13) of embodiment 10 p-methylphenyls-2-deoxidation-2-nitrine-3-O-benzyl-4-deoxidation-4-(2,2,2-trichlorine ethoxy carbonyl carboxamido-group)-6-deoxidation-6-nitrine-1-sulphur-alpha-D-glucose glycosides

2.96 gram (6.94 mmole) 12 (embodiment 9 preparations) is dissolved in 60 milliliters of anhydrous methylene chlorides, adds 2.4 milliliters of pyridines, turn on agitator slowly splashes into 2.4 milliliters of trifluoromethanesulfanhydride anhydrides with syringe at normal temperatures, and whole adition process continues 2 hours.Adding continued stirred 2 hours.Then reaction solution is poured in 150 milliliters of frozen water and stir, add sodium pyrosulfate to water layer and show slightly acidic.Tell organic layer, water layer is used an amount of washed with dichloromethane, merges organic layer, adds anhydrous sodium sulfate drying.Filter, filtrating is with revolving the steaming evaporate to dryness, and residue solidifies for a moment.With 30 milliliters of anhydrous tetrahydro furan dissolved residues, change a tube sealing then over to, the ammonia methanol solution that in tube sealing, injects 10 milliliters of 7N seals then.50 ℃ of insulation tube sealings were opened after 48 hours, sucking-off reaction solution evaporate to dryness.Residue separates with chromatographic column, obtains the substitution product and the initial raw polyol of 0.28 gram of 1.55 gram ammonia.The ammonia substitution product is dissolved in 30 milliliters of THFs, adds 1.22 gram sodium hydrogencarbonates and 1.55 gram trichlorine ethoxy dicarbonyl chlorides, normal temperature stirred 4 hours down, and processing reaction gets product 2.07 grams (3.44 mmole), and total recovery 55%, product characters are that white is gluey.

¹H?NMR(500MHz，CDCl ₃)δ7.42-7.31(m，7H)，7.15(d，J＝8.0Hz，2H)，5.55(d，J＝5.0Hz，1H)，4.90(d，J＝11.0Hz，2H)，4.77-4.66(m，3H)，4.49-4.46(m，1H)，3.96(dd，J1＝5.5Hz，J2＝10.0Hz，1H)，3.76(t，J＝10.0Hz，1H)，3.65(t，J＝10.0Hz，1H)，3.44-3.36(m，2H)，2.34(s，3H). ¹³C?NMR(125MHz，CDCl ₃)δ154.10，138.39，137.01，132.66，130.04，128.76，128.65，128.40，128.29，95.19，87.30，77.13，74.96，74.62，71.13，64.47，54.55，51.56，21.12.Anal.Calcd?for?C ₂₃H ₂₄Cl ₃N ₇O ₄S：C，45.97；H，4.03；N，16.32.Found：C，46.12；H，4.16；N，16.04.ESI-MS：Calcd.For?C ₂₃H ₂₄Cl ₃N ₇O ₄SNa([M+Na] ⁺)：622，Found622.

Embodiment 114 '-deoxidation-4 '-(2,2,2-trichlorine ethoxy carbonyl carboxamido-group)-5,3 ', 2 ", 4 ", 6 "-preparation (14) of five-O-benzyl-five azide kanendomycin

With 180 milligrams of (0.3 mmole) donors 13 (embodiment 10 preparations); 300 milligrams of (0.394 mmole) acceptors 7; 88 milligrams of (0.391 mmole) iodo succimides, 400 millimoles sieve and 5 milliliters of anhydrous methylene chlorides mix in two neck round-bottomed flasks of one 25 milliliters, are cooled to-70 ℃ under the nitrogen protection; Stir after 30 minutes, add 10 μ L trifluoromethanesulfonic acids.Slowly be warming up to-30 ℃, kept 30 minutes, slowly be warming up to room temperature then.Filter, molecular sieve is used an amount of washed with dichloromethane, will filtrate and washings merging back evaporate to dryness.Residue separates with silica gel column chromatography, with sherwood oil and ethyl acetate mixture do gradient elution (petrol ether/ethyl acetate=10: 1-5: 1).The evaporate to dryness elutriant gets 134 milligrams of α configuration products (0.108 mmole, 36%), gets 77 milligrams of beta comfiguration by products (0.062 mmole, 21%), and two kinds of product characters are the white glue.α configuration product 14: ¹H NMR (500MHz, CDCl ₃) δ 7.43-7.19 (m, 20H), 7.13-7.09 (m, 3H), 6.95-6.93 (m, 2H), 5.60 (d, J=3.5Hz, 1H), 5.56 (d; J=3.5Hz, 1H), 5.00 (d, J=11.5Hz, 1H), 4.94 (d, J=12.0Hz, 1H), 4.83-4.71 (m, 5H); 4.66 (d, J=12.0Hz, 1H), 4.61 (d, J=11.0Hz, 1H), 4.47 (d, J=12.0Hz, 1H), 4.35-4.32 (m; 1H), 4.26 (d, J1=12.0Hz, 1H), 4.23 (d, J1=11.0Hz, 1H), 3.89 (t, J=10.0Hz; 1H), and 3.81-3.77 (m, 2H), 3.70-3.61 (m, 3H), 3.58-3.27 (m, 10H), 3.14 (dd, J1=3.0Hz; J2=11.0Hz, 1H), 2.38 (ddd, J1=J2=4.5Hz, J3=13.0Hz, 1H), 1.63 (ddd, J1=J2=J3=12.0Hz, 1H). ¹³C NMR (125MHz, CDCl ₃) δ 154.02,137.85,137.59,137.34,137.26,137.02,128.55,128.52,128.32,128.18; 128.15,128.11,127.72,127.48,127.21,125.81,97.14,95.93,82.93,77.83; 77.77,77.48,76.15,75.88,75.04,74.62,74.51,74.45,73.51,73.13; 70.79,70.12,67.75,65.32,63.42,60.18,59.31,54.09,51.58,31.92.Anal.Calcd forC ₅₆H ₅₉Cl ₃N ₁₆O ₁₁: C, 54.31; H, 4.80; N, 18.09.Found:C, 54.59; H, 4.91; N, 17.88.ESI-MS:Calcd.For C ₅₆H ₅₉Cl ₃N ₁₆O ₁₁Na ([M+Na] ⁺): 1259, Found 1259.

Embodiment 125,3 ', 2 ", 4 ", 6 "-preparation (15) of five-O-benzyl-4 '-deoxidation-4 '-amino-five azide kanendomycin

480 milligram 14 (embodiment 11 preparations) are dissolved in 10 milliliter 1, and the 4-dioxane adds 1.5 ml waters, 100 milligrams of sodium hydroxide, and reflux is 3 hours under stirring.Be cooled to room temperature, reaction mixture is poured into 150 ml waters, with dichloromethane extraction three times, each 30 milliliters.Add an amount of anhydrous sodium sulfate drying extraction liquid, evaporate to dryness after filtering, residue is gone up appearance with an amount of methylene dichloride dissolving back and is walked chromatographic column, with petrol ether/ethyl acetate from 5/1 to 1/1 gradient elution, gets 262 milligrams of products, productive rate 64%, product characters is that white is gluey.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.23(m，16H)，7.22-7.19(m，4H)，7.14-7.08(m，3H)，6.96-6.93(m，2H)，5.61(d，J＝3.5Hz，1H)，5.56(d，J＝4.0Hz，1H)，5.03(d，J＝12.0Hz，1H)，4.94(d，J＝12.0Hz，1H)，4.91(d，J＝11.0Hz，1H)，4.81(d，J＝11.5Hz，1H)，4.74(d，J＝11.5Hz，1H)，4.62(d，J＝11.0Hz，1H)，4.60(d，J＝10.5Hz，1H)，4.47(d，J＝12.0Hz，1H)，4.25(d，J＝12.5Hz，1H)，4.22(d，J＝11.5Hz，1H)，3.99-3.96(m，1H)，3.81-3.77(m，2H)，3.71-3.53(m，6H)，3.49-3.44(m，2H)，3.39(dd，J1＝3.5Hz，J2＝10.5Hz，1H)，3.35(t，J＝10.0Hz，1H)，3.29(dd，J1＝2.0Hz，J2＝11.0Hz，1H)，3.21(dd，J1＝4.0Hz，J2＝10.0Hz，1H)，3.13(dd，J1＝2.5Hz，J2＝11.0Hz，1H)，2.70(t，J＝10.0Hz，1H)，2.37(ddd，J1＝J2＝4.5Hz，J3＝13.5Hz，1H)，1.61(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ137.87，137.58，137.35，137.32，128.64，128.52，128.31，128.28，128.18，128.10，127.74，127.46，127.15，125.84，97.54，95.95，83.04，80.92，77.87，77.48，75.89，75.20，75.00，74.44，73.51，73.11，73.03，70.09，67.74，65.32，63.69，60.24，59.49，54.11，51.83，32.05.Anal.Calcd?for?C ₅₃H ₅₈N ₁₆O ₉：C，59.88；H，5.50；N，21.08.Found：C，59.71；H，5.36；N，20.99.ESI-MS?Calcd?for?C ₅₃H ₅₉N ₁₆O ₉([M+H] ⁺)：1063.，Found：1063.

Embodiment 135,3 ', 2 ", 4 ", 6 "-preparation (16) of five-O-benzyl-4 '-deoxidation-4 '-dimethylamino-five azide kanendomycin

64 milligrams of (0.060 mmoles) 15 (embodiment 12 preparations) are dissolved in 3 milliliters of dry DMF, add 20 milligram of 60% sodium hydride, stir moments later, splash into 0.02 milliliter of methyl iodide, continue to stir 4 hours, and TLC shows that raw material remains in a large number.Add 40 milligrams of sodium hydrides again, 0.02 milliliter of methyl iodide, stirred overnight.TLC shows the amine residue that still has a great deal of, so add 40 milligrams of sodium hydrides again, 0.02 milliliter of methyl iodide continues to stir 5 hours, and TLC shows trace amine residue.Reaction solution is poured in 50 milliliters of salt solutions, with ethyl acetate extraction three times, each 15 milliliters.Combining extraction liquid, drying, evaporate to dryness, residue separates through chromatographic column, with petrol ether/ethyl acetate (petrol ether/ethyl acetate V/V=8: 1-5: 1) gradient elution gets 33 milligrams of products (0.030 mmole), productive rate 50%, product characters is that white is gluey.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.18(m，15H)，7.11-7.07(m，3H)，6.95-6.93(m，2H)，5.60(d，J＝3.5Hz，1H)，5.50(d，J＝4.0Hz，1H)，5.02(d，J＝11.5Hz，1H)，4.93(d，J＝11.5Hz，1H)，4.83-4.72(m，4H)，4.59(d，J＝11.0Hz，1H)，4.46(d，J＝12.0Hz，1H)，4.37-4.33(m，1H)，4.24(d，J＝12.0Hz，1H)，4.21(d，J＝11.0Hz，1H)，4.03(t，J＝10.0Hz，1H)，3.80-3.76(m，2H)，3.68-3.63(m，2H)，3.59-3.45(m，5H)，3.39(dd，J1＝3.5Hz，J2＝10.5Hz，1H)，3.35(t，J＝10.0Hz，1H)，3.28(dd，J1＝2.0Hz，J2＝11.0Hz，1H)，3.24(dd，J1＝4.0Hz，J2＝10.0Hz，1H)，3.10(dd，J1＝11.0Hz，J2＝2.5Hz，1H)，2.64(t，J＝10.0Hz，1H)，2.47(s，6H)，2.40(ddd，J1＝J2＝4.5Hz，J3＝13.5Hz，1H)，1.66(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ137.88，137.71，137.59，137.34，137.30，128.52，128.37，128.31，128.26，128.19，128.10，127.97，127.73，127.45，127.11，125.78，97.35，95.97，83.00，77.85，77.47，75.88，75.49，75.02，74.43，73.51，73.10，70.11，70.07，67.72，65.34，65.13，65.04，60.24，59.44，52.67，41.95，32.10，29.68.HRESI-MS?Calcdfor?C ₅₅H ₆₃N ₁₆O ₉([M+H] ⁺)：1091.4958，Found：1091.4960.

Embodiment 145,3 ', 2 ", 4 ", 6 "-preparation (17) of five-O-benzyl-4 '-deoxidation-4 '-acetylaminohydroxyphenylarsonic acid five azide kanendomycins

54 milligram 15 (embodiment 12 preparations) added 1.5 milliliters of pyridine mixtures stirring at room of 1 milliliter of aceticanhydride 6 hours.Reaction solution is poured in 20 milliliters of salt solutions,, use 5 milliliters in ETHYLE ACETATE at every turn with ethyl acetate extraction three times.Extraction liquid filters the back evaporate to dryness through anhydrous sodium sulfate drying, and residue is crossed a short silica gel chromatographic column, with petrol ether/ethyl acetate from 4/1 to 1/1 gradient elution, gets 45 milligrams of white gluey products, yield 80%.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.19(m，15H)，7.15-7.09(m，3H)，6.96-6.94(m，2H)，5.60(d，J＝3.5Hz，1H)，5.58(d，J＝3.5Hz，1H)，5.06(d，J＝8.5Hz，1H)，4.99(d，J＝11.5Hz，1H)，4.94(d，J＝11.5Hz，1H)，4.81(d，J＝11.5Hz，1H)，4.77(d，J＝11.5Hz，1H)，4.74(d，J＝11.5Hz，1H)，4.61(d，J＝11.0Hz，1H)，4.51(d，J＝11.5Hz，1H)，4.47(d，J＝12.0Hz，1H)，4.25(d，J＝12.5Hz，1H)，4.23(d，J＝11.0Hz，1H)，4.17-4.13(m，1H)，3.89-3.77(m，4H)，3.71(t，J＝9.5Hz，1H)，3.65(t，J＝9.5Hz，1H)，3.58-3.47(m，3H)，3.40-3.34(m，3H)，3.31-3.27(m，3H)，3.14(dd，J1＝2.5Hz，J2＝11.0Hz，1H)，2.36(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.76(s，3H)，1.61(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ170.16，137.83，137.57，137.38，137.32，137.19，128.66，128.50，128.31，128.17，128.09，127.72，127.46，127.22，125.85，96.93，95.92，82.99，77.76，77.65，77.47，76.22，75.86，75.02，74.44，73.75，73.50，73.10，71.66，70.10，67.71，65.30，63.21，60.23，59.51，51.94，51.65，31.99，23.29.HRESI-MS?Calcd?for?C ₅₅H ₆₁N ₁₆O ₁₀([M+H] ⁺)：1105.4751，Found：1105.4746.

Embodiment 155,3 ', 2 ", 4 ", 6 "-preparation (18) of five-O-benzyl-4 '-deoxidation-4 '-(4-nitrine butyrylamino)-five azide kanendomycins

85 milligrams of (0.080 mmoles) 15 (embodiment 12 preparations) are dissolved in 4 milliliters of THFs, add 300 milligrams of sodium hydrogencarbonates, splash into 0.01 milliliter of 4-chlorobutanoylchloride under stirring, and stirring at normal temperature is after 4 hours, and TLC shows the reaction completion.Reaction solution is poured in 60 milliliters of salt solutions, stirred after 2 hours with ethyl acetate extraction 3 times each 15 milliliters.Combining extraction liquid, drying, evaporate to dryness, residue are dissolved in 4 milliliters of DMF, add 20 milligrams of sodium azides, are heated to 80 ℃ and stir 6 hours, and TLC shows the reaction completion.Reaction solution is poured in 60 milliliters of salt solutions, fully stirred, with ethyl acetate extraction 3 times, each 15 milliliters.Combining extraction liquid, drying, evaporate to dryness, residue separates through a shorter chromatogram column, with petrol ether/ethyl acetate (petrol ether/ethyl acetate V/V=5: 1-2: 1) gradient elution, 75 milligrams of products (0.064 mmole), two step yields 80%, product characters be white glue.

¹H?NMR(500MHz，CDCl ₃)δ7.42-7.11(m，23H)，7.12-7.00(m，4H)，6.95-6.94(m，2H)，5.60(d，J＝3.5Hz，1H)，5.58(d，J＝3.5Hz，1H)，5.13(d，J＝8.5Hz，1H)，5.00-4.93(m，2H)，4.82-4.69(m，3H)，4.61(d，J＝11.0Hz，1H)，4.51(d，J＝11.5Hz，1H)，4.47(d，J＝12.0Hz，1H)，4.25(d，J＝12.0Hz，1H)，4.23(d，J＝11.0Hz，1H)，4.18-4.14(m，1H)，3.93-3.77(m，4H)，3.71(t，J＝9.5Hz，1H)，3.65(t，J＝9.5Hz，1H)，3.59-3.48(m，3H)，3.42-3.34(m，3H)，3.32-3.25(m，4H)，3.16-3.13(m，2H)，2.37(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，2.06-1.96(m，2H)，1.82-1.76(m，2H)，1.61(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，CDCl ₃)δ171.87，137.83，137.57，137.37，137.32，137.19，128.64，128.51，128.44，128.31，128.17，128.14，128.09，127.90，127.73，127.46，127.23，125.84，96.96，95.91，82.97，77.73，77.47，76.25，75.86，75.01，74.44，73.64，73.50，73.11，71.60，70.10，67.72，65.31，63.12，60.21，59.49，51.95，51.52，50.66，32.91，31.98，24.33，13.80.HRESI-MS?Calcd?for?C ₅₇H ₆₃N ₁₉O ₁₀Na([M+Na] ⁺)：1196.4898，Found：1196.4894.

Embodiment 165,3 ', 2 ", 4 ", 6 "-preparation (19) of five-O-benzyl-4 '-deoxidation-4 '-hexamethylene carbonyl carboxamido-group-five azide kanendomycin

16 milligrams of heptanaphthenic acid (0.122 mmole) of 65 milligrams of (0.061 mmoles) 15 (embodiment 12 preparations), 4 milliliters of N, dinethylformamide; 3 N; The N-diisopropyl ethyl amine, 47 milligrams of (0.17 mmole) benzotriazole tetrafluoride boron salt join in the flask successively, stir 12 hours.Processing reaction, crude product separates with silica gel chromatographic column, with petrol ether/ethyl acetate from 4/1 to 1/1 gradient elution, gets 56 milligrams of products (0.044 mmole), productive rate 78%, product characters is white jelly.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.19(m，20H)，7.14-7.08(m，3H)，6.95-6.94(m，2H)，5.60(d，J＝3.5Hz，1H)，5.59(d，J＝4.0Hz，1H)，5.20(d，J＝8.0Hz，1H)，4.99(d，J＝12.0Hz，1H)，4.94(d，J＝12.0Hz，1H)，4.81(d，J＝12.0Hz，1H)，4.74(d，J＝11.5Hz，1H)，4.71(d，J＝11.5Hz，1H)，4.61(d，J＝11.0Hz，1H)，4.53(d，J＝11.5Hz，1H)，4.47(d，J＝12.0Hz，1H)，4.25(d，J＝12.0Hz，1H)，4.22(d，J＝11.0Hz，1H)，4.18-4.15(m，1H)，3.95-3.86(m，2H)，3.81-3.77(m，2H)，3.72(t，J＝9.5Hz，1H)，3.65(d，J＝9.5Hz，1H)，3.58-3.48(m，3H)，3.40-3.34(m，3H)，3.31-3.24(m，3H)，3.14(dd，J1＝2.5Hz，J2＝11.0Hz，1H)，2.36(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.86-1.80(m，1H)，1.74-1.57(m，6H)，1.37-1.14(m，5H). ¹³C?NMR(125MHz，CDCl ₃)δ176.14，137.84，137.57，137.33，137.28，137.20，128.51，128.52，128.31，128.17，128.14，128.10，127.73，127.46，127.22，125.86，97.01，95.91，83.00，77.76，77.47，76.25，75.86，75.01，74.44，73.58，73.50，73.10，71.76，70.09，67.71，65.31，62.97，60.23，59.53，51.98，51.25，45.31，32.01，29.68，29.18，25.58，25.53，25.48.HRESI-MS?Calcd?for?C ₆₀H ₆₈N ₁₆O ₁₀Na([M+Na] ⁺)：1195.5197，Found：1195.5177.

Embodiment 174 '-deoxidation-4 '-[2-(2-toluyl amido)-kharophen]-5,3 ', 2 ", 4 ", 6 "-preparation (20) of five-oxygen-benzyl-five azide kanendomycin

Compound method is with 19 (embodiment 16).By 65 milligrams of (0.061 mmoles) 15, get 57 milligrams of products (0.046 mmole), yield 75%, product characters is a white crystalline powder.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.18(m，24H)，7.13-7.08(m，3H)，6.95-6.93(m，2H)，6.60(d，J＝8.5Hz，1H)，6.48(t，J＝4.5Hz，1H)，5.61(d，J＝3.5Hz，1H)，5.51(d，J＝3.5Hz，1H)，5.02(d，J＝12.0Hz，1H)，4.92(d，J＝12.0Hz，1H)，4.82-4.73(m，3H)，4.60(d，J＝11.0Hz，1H)，4.53(d，J＝11.5Hz，1H)，4.46(d，J＝12.0Hz，1H)，4.30-4.21(m，3H)，4.00-3.85(m，3H)，3.81-3.73(m，3H)，3.65-3.61(m，2H)，3.52-3.46(m，2H)，3.40-3.22(m，7H)，3.12(dd，J1＝3.0Hz，J2＝11.0Hz，1H)，2.45(s，3H)，2.27(ddd，J1＝J2＝4.5Hz，J3＝13.0Hz，1H)，1.58(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³CNMR(125MHz，CDCl ₃)δ170.36，168.80，137.84，137.71，137.56，137.32，136.47，147.75，131.23，130.52，128.51，128.46，128.30，128.26，128.17，128.09，127.91，127.73，127.70，127.44，127.20，127.13，125.88，125.72，97.28，95.90，82.87，78.06，77.71，77.44，77.09，75.86，75.02，74.42，74.24，73.49，73.10，70.99，70.06，67.71，65.32，63.35，60.17，59.04，51.88，51.65，43.84，31.79，20.05.HRESI-MS?Calcd?for?C ₆₃H ₆₇N ₁₇O ₁₁Na([M+Na] ⁺)：1260.5098，Found：1260.5088.

Embodiment 185,3 ', 2 ", 4 ", 6 "-preparation (21) of amino-five azide kanendomycins of five-O-benzyl-4 '-deoxidation-4 '-[(N-carbobenzoxy-(Cbz)-piperidyl)-4-carbonyl]

Compound method is with 19 (embodiment 16).By 65 milligrams of (0.061 mmoles) 15, get 65 milligrams of products (0.050 mmole), 81% yield, product characters is that white is gluey.

¹H?NMR(500MHz，CDCl ₃)δ7.43-7.09(m，28H)，6.96-6.93(m，2H)，5.60(d，J＝3.5Hz，1H)，5.59(d，J＝3.5Hz，1H)，5.27(d，J＝8.0Hz，1H)，5.11(s，2H)，4.98(d，J＝12.0Hz，1H)，4.94(d，J＝12.0Hz，1H)，4.81(d，J＝12.0Hz，1H)，4.74(d，J＝12.0Hz，1H)，4.73(d，J＝11.5Hz，1H)，4.61(d，J＝11.0Hz，1H)，4.49(d，J＝12.0Hz，1H)，4.47(d，J＝12.0Hz，1H)，4.25(d，J＝12.0Hz，1H)，4.23(d，J＝11.0Hz，1H)，4.17-4.14(m，3H)，3.94-3.84(m，2H)，3.81-3.77(m，2H)，3.72(t，J＝9.5Hz，1H)，3.65(t，J＝9.5Hz，1H)，3.58-3.48(m，3H)，3.40-3.28(m，5H)，3.24(dd，J1＝2.5Hz，J2＝13.0Hz，1H)，3.14(dd，J1＝3.0Hz，J2＝11.0Hz，1H)，2.73(br，2H)，2.38-2.32(m，1H)，1.96-1.90(m，1H)，1.70-1.57(m，3H)，1.56-1.46(m，2H). ¹³C?NMR(125MHz，CDCl ₃)δ174.28，155.04，137.82，137.55，137.30，137.16，136.62，128.58，128.49，128.30，128.15，128.07，127.86，127.70，127.45，127.22，125.83，96.95，95.88，82.95，77.76，77.70，77.46，76.30，75.84，74.99，74.42，73.48，73.09，71.65，70.09，67.71，67.14，65.29，62.95，60.19，59.50，51.93，51.25，43.21，43.16，42.85，31.97，28.38，27.99.HRESI-MS?Calcd?for?C ₆₇H ₇₃N ₁₇O ₁₂Na([M+Na] ⁺)：1330.5517，Found：1330.5516.

The preparation of embodiment 194 '-deoxidation-4 '-dimethylamino-kanendomycin (22)

33 milligrams of (0.030 mmoles) 16 (embodiment 13 preparations) are dissolved in the mixture of 3 milliliters of pyridine/2 milliliter triethylamines, and logical hydrogen sulfide to reaction solution is deep green.Stirred 4 hours; Solvent evaporated, residue are adsorbed onto on the proper silica gel after dissolving with 2 milliliters of methylene dichloride, forward a little chromatographic column behind the evaporate to dryness to; Press following sequentially eluting liquid wash-out: 50 milliliters of ETHYLE ACETATE; 50 milliliters of ethyl acetate/methanol (V/V=1/1), 50 ml methanol, 50 ml methanol/strong aqua (V/V=10/1).Collection contains the part of product, evaporate to dryness.Crude product dissolves with 5 ml methanol, transfers pH to 3-4 with the Hydrogen chloride of 1N, adds the palladium carbon of catalytic amount, feeds hydrogen hydrogenation 5 days, and TLC shows that single product generates.Remove by filter insolubles, the filtrating evaporate to dryness, residue dissolves with 5 ml waters, removes freeze-drying behind the insolubles, must 21 milligrams of products (0.029 mmole), yield 95%, white amorphous powder.

¹H?NMR(500MHz，D ₂O)δ6.09(d，J＝4.0Hz，1H)，5.15(d，J＝4.0Hz，1H)，4.66(dt，J1＝3.0Hz，J2＝J3＝10.0Hz，1H)，4.58(t，J＝10.5Hz，1H)，4.16(t，J＝10.0Hz，1H)，3.98-3.92(m，3H)，3.90-3.86(m，2H)，3.82-3.61(m，7H)，3.54-3.46(m，2H)，3.12(s，6H)，2.56(ddd，J1＝4.0Hz，J2＝J3＝12.5Hz，1H)，2.02(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，D ₂O)δ101.37，96.34，84.37，79.03，74.83，73.74，68.74，67.13，66.14，65.48，63.22，60.66，55.56，55.02，50.25，48.68，43.14，41.06，28.56.HRESI-MS?Calcd?forC ₂₀H ₄₃N ₆O ₉([M+H] ⁺)：511.3086，Found：511.3067.

The preparation (23) of embodiment 204 '-deoxidation-4 '-acetylaminohydroxyphenylarsonic acid kanendomycin

Deprotection method is the same.By 45 milligrams of (0.041 mmoles) 17 (embodiment 14 preparations), obtain 25 milligrams of products (0.035 mmole), yield 87%, white amorphous powder.

¹H?NMR(500MHz，D ₂O)δ6.06(d，J＝3.5Hz，1H)，5.14(d，3.5Hz，1H)，4.25(t，J＝10.0Hz，1H)，4.20-4.16(m，1H)，4.09(t，J＝10.0Hz，1H)，3.98-3.93(m，3H)，3.89-3.77(m，4H)，3.72(t，J＝10.0Hz，1H)，3.68-3.59(m，2H)，3.55(dd，J1＝4.0Hz，J2＝10.5Hz，1H)，3.52(t，J＝10.5Hz，1H)，3.36(dd，J1＝2.5Hz，J2＝14.0Hz，1H)，3.16(dd，J1＝6.5Hz，J2＝14.0Hz，1H)，2.58-2.55(m，1H)，2.08(s，3H)，2.02(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，D ₂O)δ176.11，101.39，96.48，84.44，77.72，75.04，73.66，68.87，68.80，66.13，60.63，55.60，54.84，53.27，50.33，49.09，40.96，28.53，22.78.HRESI-MS?Calcdfor?C ₂₀H ₄₁N ₆O ₁₀([M+H] ⁺)：525.2879，Found：525.2865.

The preparation (24) of embodiment 214 '-deoxidation-4 '-(4-amino-butyrylamino)-kanendomycin

Deprotection method is the same.By 66 milligrams of (0.056 mmoles) 18 (embodiment 15 preparations), get 29 milligrams of products (0.037 mmole), yield 66%, white amorphous powder.

¹H?NMR(500MHz，D ₂O)δ6.04(d，J＝3.5Hz，1H)，5.14(d，3.5Hz，1H)，4.23(t，J＝10.5Hz，1H)，4.21-4.18(m，1H)，4.08(t，J＝9.0Hz，1H)，3.98-3.50(m，12H)，3.36(dd，J1＝3.0Hz，J2＝14.0Hz，1H)，3.16(dd，J1＝5.5Hz，J2＝13.5Hz，1H)，3.04(t，J＝7.5Hz，2H)，2.58-2.55(m，1H)，2.49(t，J＝7.5Hz，2H)，2.03-1.94(m，3H). ¹³C?NMR(125MHz，D ₂O)δ176.80，101.42，96.75，84.46，78.08，74.98，73.70，68.92，68.76，66.13，60.61，55.58，54.91，53.06，50.28，49.05，40.89，39.59，33.10，28.54，23.40.HRESI-MS?Calcd?forC ₂₂H ₄₆N ₇O ₁₀([M+H] ⁺)：568.3301，Found：568.3300.

The preparation (25) of embodiment 224 '-deoxidation-4 '-hexamethylene carbonyl carboxamido-group-kanendomycin

Deprotection method is the same.By 46 milligrams of (0.074 mmoles) 19 (embodiment 16 preparations), obtain 35 milligrams of products (0.045 mmole), yield 61%, white amorphous powder.

¹H?NMR(500MHz，D ₂O)δ6.04(d，J＝4.0Hz，1H)，5.14(d，J＝3.5Hz，1H)，4.22(t，J＝10.0Hz，1H)，4.20-4.17(m，1H)，4.07(t，J＝10.0Hz，1H)，3.99-3.78(m，7H)，3.73(t，J＝10.0Hz，1H)，3.69-3.60(m，3H)，3.57-3.50(m，2H)，3.32(dd，J1＝2.5Hz，J2＝13.5Hz，1H)，3.11(dd，J1＝5.5Hz，J2＝14.0Hz，1H)，2.59-2.55(m，1H)，2.37-2.31(m，1H)，2.02-1.95(m，1H)，1.84-1.75(m，4H)，1.67(d，J＝12.0Hz，1H)，1.43-1.14(m，5H). ¹³C?NMR(125MHz，D ₂O)δ182.22，101.43，96.85，84.48，78.12，74.99，73.72，68.99，68.75，66.08，63.27，60.60，55.59，54.98，52.69，50.27，49.05，45.68，40.79，29.91，28.56，25.93，25.80.HRESI-MS?Calcd?for?C ₂₅H ₄₉N ₆O ₁₀([M+H] ⁺)：593.3505，Found：593.3504.

The preparation (26) of embodiment 234 '-deoxidation-4 '-[2-(2-toluyl amido)-kharophen]-kanendomycin

Deprotection method is the same.By 46 milligrams of (0.037 mmoles) 20 (embodiment 17 preparations), get 20 milligrams of products (0.023 mmole), yield 64%, light yellow amorphous powder.

¹H?NMR(500MHz，D ₂O)δ7.47-7.42(m，2H)，7.35-7.29(m，2H)，6.07(d，J＝3.5Hz，1H)，5.14(d，J＝3.0Hz，1H)，4.31(t，J＝10.0Hz，1H)，4.29-4.25(m，1H)，4.18-4.08(m，3H)，3.97-3.83(m，6H)，3.79(dd，J1＝5.0Hz，J2＝12.5Hz，1H)，3.72(t，J＝10.0Hz，1H)，3.70-3.61(m，2H)，3.59-3.50(m，2H)，3.40(d，J＝12.0Hz，1H)，3.20(dd，J1＝6.0Hz，J2＝13.5Hz，1H)，2.58-2.56(m，1H)，2.37(s，3H)，2.01(ddd，J1＝J2＝J3＝12.5Hz，1H). ¹³C?NMR(125MHz，D ₂O)δ174.76，173.29，136.59，135.15，131.66，131.49，127.83，126.65，101.42，96.65，84.47，77.91，75.03，73.71，68.79，66.12，65.98，60.63，55.60，54.88，53.46，50.31，49.09，43.77，40.91，28.55，19.44.HRESI-MS?Calcd?for?C ₂₈H ₄₈N ₇O ₁₁([M+H] ⁺)：658.3406，Found：658.3413.

The preparation (27) of embodiment 244 '-deoxidation-4 '-(piperidines-4-carbonyl) amino-kanendomycin

Deprotection method is the same.By 53 milligrams of (0.041 mmoles) 21 (embodiment 18 preparations), get 19 milligrams of products (0.024 mmole), yield 60%, white amorphous powder.

¹H?NMR(500MHz，D ₂O)δ6.08(d，J＝4.0Hz，1H)，5.14(d，J＝3.5Hz，1H)，4.28(t，J＝10.5Hz，1H)，4.24-4.17(m，1H)，4.09(t，J＝10.0Hz，1H)，3.97-3.82(m，5H)，3.78(dd，J1＝5.0Hz，J2＝12.5Hz，1H)，3.71(t，J＝10.0Hz，1H)，3.67-3.59(m，3H)，3.55-3.44(m，4H)，3.33-3.26(m，1H)，3.18(dd，J1＝7.0Hz，J2＝14.5Hz，1H)，3.13-3.05(m，1H)，2.77-2.72(m，1H)，2.56-2.54(m，1H)，2.10-1.98(m，2H)，1.91-1.83(m，2H)，1.58-1.56(m，1H). ¹³C?NMR(125MHz，D ₂O)δ178.18，101.42，96.61，84.47，77.84，75.03，73.70，68.89，68.79，66.12，66.08，60.63，55.61，54.92，53.02，50.31，49.10，43.81，43.76，40.97，40.67，28.55，25.91，25.60.HRESI-MS?Calcd?for?C ₂₄H ₄₈N ₇O ₁₀([M+H] ⁺)：594.3457，Found：594.3453.

It is to be noted that synthetic route of the present invention selected more typical side chain for use to C4 ' deutero-aminoglycoside; As R during for straight chain amino/carboxamido-group of only containing carbochain or substitution in side chain amino/carboxamido-group; Those of ordinary skill in the art is after having read this specification sheets; Be easy to obtain the carbochain (for example 4-18 carbon atom) of the substituting group different lengths of N atom according to the working method of present embodiment; Similar; Also can obtain other the amino that contains Sauerstoffatom, nitrogen-atoms/carboxamido-group side chain fully and can select the carbochain (for example 2-18 carbon atom) of different lengths, it is all attainable that these all are that those skilled in the art need not to pay any performing creative labour, so protection scope of the present invention never is limited to the disclosed scope of present embodiment.

Test Example 1 aminoglycoside compounds and pharmaceutically useful salt anti-microbial activity thereof involved in the present invention tested

One, test materials and source

1, the compound that test compound: embodiment of the invention 19-24 is prepared, numbering is respectively 22-27; And with kanamycin A that compares and the commercially available article of kanendomycin.

2, experiment material: non-resistant organism: ATCC 25922, ATCC 29213, and ATCC 12228, ATCC35218; Resistant organism: Klebsiellapneumoniae (ATCC 700603, expression ANT (2 ")), Pseudomonasaeruginosa (ATCC 27853; express APH (3 ')-IIb), the Staphylococcus aureus of methicillin resistance (ATCC 33591, express APH (3 '); ANT (4 '), and AAC (6 ')/APH (2 ")), (ATCC 29212 for Enterococcus faecalis; Express efflux pump), Mortierella alpine (ATCC32221, fungi); The Staphylococcus epidermidis 08-18 of methicillin resistance (available from biotechnology research institute of the Chinese Academy of Medical Sciences), the Staphylococcus epidermidis 07-9 of methicillin resistance (available from biotechnology research institute of the Chinese Academy of Medical Sciences) and Escherichiacoli BL21 (pET29a) (available from biotechnology research institute of the Chinese Academy of Medical Sciences).Staphylococcus epidermidis08-18 and Staphylococcus epidermidis 07-9 are isolating clinically methicillin resistance strains; E.coli BL21 (pET29a) is that the persister of reorganization (contains the pET29a expression vector, can express APH (3 ')-Ia).

Two, TP

The mensuration of minimum inhibitory concentration (MIC) is according to Standardization Research institute of U.S. clinical labororatory (Clinical andLaboratory Standards Institute, CLSI) operational guidance of promulgation enforcement (NationalCommittee for Clinical Laboratory Standards.2006.Methods for dilutionantimicrobial susceptibility tests for bacteria that growaerobically; Approved standard---seventh edition.NationalCommittee forClinical Laboratory Standards, Wayne, Pa.).

(1), plant bacterium:

1. from-80 ℃ of Ultralow Temperature Freezers, take out the bacterial classification of preserving, be inoculated in respectively in the 1ml MH meat soup.

2.37 ℃ incubated overnight (16-18 hour) promptly can be used for the MIC experiment.

(2), mother liquid medicine preparation:

Use the sterilized water compounding pharmaceutical, making its concentration is that the medicine for preparing of 2560ug/ml. is subsequent use as for-20 ℃.

(3), drug dilution liquid preparation:

1. prepare the test tube of 12 sterilizations, and numbering, first test tube adds 8ml MH meat soup, and all the other test tubes all add 4mlMH meat soup.

2. first test tube takes out 400 microlitres, and abandons.The mother liquid medicine of getting 400 microlitre 2560ug/ml adds in first test tube, obtains the medicament solution of 128ug/ml.Then, the medicament solution that from first test tube, takes out 4ml 128ug/ml adds in second test tube, and then the solution of second test tube is 64ug/ml. and then gets 4ml from second test tube and add the medicament solution that the 3rd test tube obtains 32ug/ml, operates equally successively to a times gradient dilution medicine; Obtain 16ug/ml respectively, 8ug/ml, 4ug/ml; 2ug/ml, 1ug/ml, 0.5ug/ml; 0.25ug/ul, 0.125ug/ul, the medicament solution of 0.06ug/ul.

3. the diluent of the different concns of each medicine all carries out according to above-mentioned steps.

(4), shop 96 orifice plates (costar 3599, and corning company produces)

1. the medicament solution with 4ml 128ug/ml adds the sterilization V-shaped groove, gets the 1st perpendicular row (both A1 to H1 holes) that 100ul adds 96 orifice plates with the 8 passage volley of rifle fires.The residue soup abandons.

2. the medicament solution with 4ml 64ug/ml adds the sterilization V-shaped groove, gets the 2nd perpendicular row (both A2 to H2 holes) that 100ul adds 96 orifice plates with the 8 passage volley of rifle fires.The residue soup abandons.

3. successively the soup of all the other concentration is added in dull and stereotyped each hole in proper order.The corresponding same drug level of each perpendicular row.

(5), dilution of bacterium liquid and bed board

1. the bacterium with cultivation overnight is diluted to 0.5 Maxwell.(bacterial concentration of 0.5 Maxwell is equivalent to 1.5 * 108cfu/ml.

2. again 15 times of the bacterium liquid of 0.5 Maxwell dilutions.Both having got 0.5ml adds in the 7.5mlMH meat soup.

3. above-mentioned bacterium liquid is added in the aseptic V-shaped groove, get 5ul with the 12 passage volley of rifle fires and add among the A1-A12.(bacterium liquid final concentration is 5 * 10 in every hole ⁵Cfu/ml)

4. other bacterium adds B successively, C, and D, E, F, among the G, every dull and stereotyped H capable (H1-H12) is set to not add the negative control of bacterium.

With flat board as for 37 ℃ of thermostatic chamber cultivations overnight, observations after 16-18 hour.

Annotate: every 96 orifice plate is a medicine, and each walks crosswise corresponding bacterium, and one flat plate can be done 7 bacterium, if the kind increase of bacterium then corresponding increase bed board quantity.Until satisfying the experiment demand.

(6) outcome record

1. each is walked crosswise, and (from A12-A1) first concentration that does not have a bacteria growing is promptly to the MIC value of should medicine corresponding corresponding bacterium from right to left.If minimum concentration 0.06ug/ml does not have bacteria growing, if then MIC were recorded as＜0.06ug/ml. maximum concentration 128ug/ml would still have bacteria growing then is recorded as＞128ug/ml.

2. every dull and stereotyped negative control has bacteria growing then test-results is invalid.

3. same test repetitive operation three times guarantees that the result is reliable and stable.

Three, test-results

From the active testing result, except dimethylamino substitution compound 22 only shows the faint activity non-resistant organism, the carboxamido-group substitution compound has all shown good anti-microbial activity.Especially compound 24 and 27 not only shows good activity to non-resistant organism, and some bacteriums of expressing the resistance enzyme have also been represented the excellent antibiotic activity; Such as the Klebsiella Pneumoniae (Klebsiella pneumoniae, ATCC 700603) of expressing ANT (2 "), express the Pseudomonas aeruginosa (Pseudomonas aeruginosa; ATCC 27853) of APH (3 ')-IIb; express the golden Portugal peach coccus (Staphylococcus aureus, ATCC 33591, express APH (3 '); ANT (4 ') and AAC (6 ')/APH (2 ")) of plurality of enzymes etc., and the staphylococcus epidermidis of methicillin resistance (Staphylococcus epidermidis08-18).(contain the pET29a expression vector, express in the test of APH (3 ')-Ia), 24 and 27 have also showed better antibacterial activity, confirm that this compound can tolerate the attack of APH (3 ')-Ia to the persister E.coli BL21 (pET29a) of reorganization.Compound 25 has also shown good anti-enzymatic property.

Above-mentioned test-results shows that the improved aminoglycoside molecule of process of the present invention has good antibacterial activity, and the resistant organism that produces enzyme is still had good bacteriostatic activity.

Claims (9)

1. the aminoglycoside compounds shown in the formula formula (I) or its pharmaceutically useful salt:

Formula (I)

R ¹Be selected from Wasserstoffatoms, substituted alkoxyl group, substituted amino or substituted amido;

R ²Be selected from Wasserstoffatoms or hydroxyl;

R ³Be selected from amino or hydroxyl;

R ⁴Be selected from Wasserstoffatoms, β-D-ribofuranosyl, 3-oxygen-(2; 6-diamino--2; 6-dideoxy-β-L-Chinese mugwort Du pyrans glycosyl)-β-D-ribofuranosyl, 3-oxygen-[4-oxygen (α-D-mannopyranose base)-2,6-diamino--2 or 6-dideoxy-β-L-Chinese mugwort Du pyrans glycosyl]-β-D-ribofuranosyl;

R ⁵Be selected from Wasserstoffatoms, 3-amino-3-deoxidation-α-D-glucopyranosyl or 3-amino-3-deoxidation-4-methyl-α-D-xylopyranosyl;

R ⁶Be selected from Wasserstoffatoms, alkyl, substituted alkyl or substituted acyl group;

R ⁷Be selected from hydroxyl, amino or methylamino;

R ⁸Be selected from Wasserstoffatoms or methyl.

2. according to the described aminoglycoside compounds of claim 1 or its pharmaceutically useful salt, it is characterized in that:

R ¹Described in substituted amino comprise dimethylamino or kharophen; Described substituted amido comprises the amino butyrylamino of 4-, piperidines-4-formamido-, cyclohexyl formamido-or 2-(2-toluyl amido) acetamido;

R ⁶Described in alkyl comprise ethyl; Described substituted acyl group comprises 2-hydroxyl-4-amino-butyryl radicals.

3. method for preparing the aminoglycoside compounds of formula shown in the described formula of claim 1 (I) comprises:

(1), obtain the glycosyl acceptor that removes ring I shown in the formula III with the aminoglycoside compound cracking shown in the formula II:

formula II

formula III

(2) glycosyl donor of 4 bit strip temporary protection bases shown in the preparation formula IV:

formula IV

(3) through glycosylation the glycosyl donor shown in glycosyl acceptor shown in the formula III and the formula IV is carried out coupling, 4 ' protection base sloughing on the formula IV compound ring I also further derived, and finally sloughs all protections, promptly gets.

4. according to the described method of claim 3, it is characterized in that: 3 ' and 4 ' hydroxyl of the ring I of the aminoglycoside compound molecule shown in step (1) the Chinese style II is exposed, other hydroxyl or amino the protection.

5. according to the described method of claim 3, it is characterized in that: with the process of the aminoglycoside compound shown in formula II sodium periodate oxidation, alkaline condition descends and separates the formula III compound that obtains as glycosyl acceptor in the step (1).

6. according to the described method of claim 3; It is characterized in that: the formula IV compound described in the step (2) exists with sulphur glycosides or tribromo-acetyl polyurethane form; 4 hydroxyl or amino to be different from other hydroxyl or the amino protection base protection on glycosyl acceptor or the donor can optionally be removed after glycosylation is accomplished.

7. according to the described method of claim 3, it is characterized in that: glycosylation described in the step (3) comprises sulphur glycosides glycosylation or the acid catalyzed trichlorine polyurethane of Lewis glycosylation that NIS/TfOH mediates.

8. pharmaceutical composition of treating infectation of bacteria is characterized in that: be made up of the claim 1 of significant quantity or 2 described aminoglycoside compounds or its pharmaceutically useful salt and pharmaceutically acceptable carrier or auxiliary material.

9. claim 1 or 2 described aminoglycoside compounds or its pharmaceutically useful salt purposes in the medicine of preparation treatment infectation of bacteria.

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