CN1636008A

CN1636008A - Macrolide antiinfective agents

Info

Publication number: CN1636008A
Application number: CNA008090726A
Authority: CN
Inventors: D·T·W·褚
Original assignee: Kosan Biosciences Inc
Current assignee: Kosan Biosciences Inc
Priority date: 1999-04-16
Filing date: 2000-04-14
Publication date: 2005-07-06
Also published as: BR0010681A; MXPA01010529A; JP2003523939A; CA2369886A1; IL145776A0; NZ514791A; EP1171448A2

Abstract

Compounds of formula (1), (2) or (3) or the 10,11-anhydro forms thereof, wherein Ra is H or OH; Rb is H or halogen; Rc is H or a protecting group; Rd is methyl; unsubstituted alkyl(3-10C); substituted alkyl(1-10C); substituted or unsubstituted alkenyl(2-10C) or substituted or unsubstituted alkynyl(2-10C); substituted or unsubstituted aryl(4-14C); substituted or unsubstituted arylalkyl(5-20C); substituted or unsubstituted arylalkenyl(5-20C); substituted or unsubstituted arylalkynyl(5-20C); substituted or unsubstituted amidoarylalkyl(5-20C); substituted or unsubstituted amidoarylalkenyl(5-20C); or substituted or unsubstituted amidoarylalkynyl(5-20C); Re is H or a protecting group or is mono- or disubstituted amino carbonyl; Rf is H; substituted or unsubstituted alkyl(1-10C), substituted or unsubstituted alkenyl(1-10C); substituted or unsubstituted alkynyl(1-10C); substituted or unsubstituted aryl(4-14C); substituted or unsubstituted arylalkyl(5-20C); or -ORf may be replaced by -H; one of Z and Y is H and the other is OH or protected OH, or is amino, mono- or dialkyl-amino, protected amino, or an aminoheterocycle or Z and Y together are =O, =NOH or a derivatized oxime; including any pharmaceutically acceptable salts thereof and any stereoisomeric forms and mixtures of stereoisomeric forms thereof, are antimicrobial agents.

Description

Macrolide antiinfective agents

Technical field

The present invention relates to enlarge the antimicrobial compounds of erythromycin sample microbiotic inventory.More specifically, the present invention relates to contain the macrolide antibiotic of the erythronolids nuclear of modifying at the substituting group place of C-13 at least.

Background technology

People recognize that acquisition increases progressively the microorganism strains quantity of the resistance of present known Antibiotique composition, are a kind of threats to public health.Along with the use of these compounds increases sharply, the selection needs that are used for the treatment of the state of an illness that microorganism of all kinds causes are also increasing.Surpass human infection's treatment for the bigger needs of Antimicrobe compound, and expanded to the needs of preserving food and other perishables.New microbiotic also is essential for resistance plant and animal, and provides protection to the material that meeting is subjected to microbiological corrosion.

Therefore, significant need enlarges the arsenal that many-sided defence undesirable microorganism active compound can be provided.

The WO98/09978 (being incorporated herein for your guidance) that published on March 12nd, 1998 discloses the modified forms of erythromycin, and they do not have cladinose in the 3-position, in all sorts of ways in the 9-12 position of macrolide ring and derive.Similarly, the U.S. Patent number of submitting on May 12nd, 1,998 5,750,510 (being incorporated herein for your guidance) discloses the erythromycin derivatives of modifying.

Naturally occurring erythromycin has structure

Erythromycin R' R" A-OH-CH ₃B-H-CH ₃C-OH-HD-H-H

Wherein R ' can be H or OH, R " can be H or CH ₃

Disclosed all compounds all have ethyl 13 of macrolide ring in the above-mentioned patent documentation of quoting.The present invention finds to change 13 substituting group and causes many excellent antibiotic active compounds that have.

Disclosure of the Invention

The present invention relates to contain erythronolids derivative from the modification of natural structure.All compounds of the present invention are all modified at 13 quilts at least.

Therefore, in one aspect, the present invention relates to the compound of following formula

Or

Or its 10, the 11-dehydrated form;

Wherein

R _aBe H or OH, preferred OH;

R _bBe H or halogen;

R _cBe H or blocking group;

R _dIt is methyl; Unsubstituted alkyl (3-10C); The alkyl (1-10C) that replaces; Replace or unsubstituted alkenyl (2-10C); Replace or unsubstituted alkynyl (2-10C); Replace or unsubstituted aryl (4-14C); Replace or unsubstituted aralkyl (5-20C); Replace or unsubstituted aromatic yl alkenyl (5-20C); Replace or unsubstituted aromatic yl polysulfide yl (5-20C); Replace or unsubstituted amido aralkyl (5-20C); Replace or unsubstituted amido aromatic yl alkenyl (5-20C); Or replacement or unsubstituted amido aromatic yl polysulfide yl (5-20C);

R _eIt is H or blocking group or single-or dibasic aminocarboxyl;

R _fBe H; Replace or unsubstituted alkyl (1-10C), replacement or unsubstituted alkenyl (1-10C); Replace or unsubstituted alkynyl (1-10C); Replace or unsubstituted aryl (4-14C); Replace or unsubstituted aralkyl (5-20C); Or-OR _fCan be replaced by-H;

One of Z and Y are H, and another is the OH of OH or protection, or amino, single-or dialkyl amido, the amino of protection, or amino-heterocycles or

Z and Y be altogether=O ,=NOH or deutero-oxime;

Comprise acceptable salt and any stereoisomeric forms in any ratio on any its pharmacology, and the mixture of stereoisomeric forms in any ratio.

In yet another aspect, the present invention relates to contain the medicine or the protection composition of the compound of formula (1)-(3), and use these compounds for treating infectious diseases, or provide their to preserve method of material.

Compound of the present invention has antibiotic activity, but be preferably used as 10 of compound, half-synthetic mesophase thing of 11 dehydrated forms, these intermediates further convert the compound with erythronolids core to, between the C10 of erythronolids core and C11 position, has ring, as the U.S. Provisional Patent Application submitted on June 18th, 1999 number 60/140,175, with 60/172 of submission on December 17th, 1999,159, with the novel number of patent application of submitting on April 14th, 2000 of U.S. utility _ _ _ _, be entitled as " macrolide anti-infection agent " (proxy number 30062-203800), be incorporated herein for your guidance.

The accompanying drawing summary

Fig. 1 has shown the synthetic schemes of The compounds of this invention.

Fig. 2 has shown that the PKS artifact of erythromycin is synthetic.As shown in Figure 1, the present invention has used this approach.

Fig. 3 has shown the synthetic of formula (3) compound, wherein R _fIt is methyl.

Fig. 4 has shown the compound of formula (1) and corresponding 10,11-dehydrated form synthetic.

Fig. 5 has shown compound synthetic of formula (3), wherein OR _fReplaced by H.

Fig. 6 has illustrated that 15-azido-Erythromycin A changes into 15-amido erythromycin.

Implement mode of the present invention

Can unite synthesising chemical technology and the synthetic easily compound of the present invention of the microbial process relevant with genetic engineering modified microorganism.Simply say, in implementing preference pattern of the present invention, provide a kind of microorganism host with recombinant expression system, preferably itself do not produce the host of macrolide antibiotic, be used to produce the 6-deoxidation erythronolide B (6-dEB) of modification, its expression system will change by the division in the ketone synthetic enzyme part catalysis territory in first assembly in some cases.For R _dBe the substituting group of methyl, with the host cell that does not have the merblastic functional domain of ketone synthetic enzyme.This change in 6-dEB polyketide synthase (PKS) makes this PKS can not utilize its natural start element, thereby can in the reaction sequence of the 6-dEB that causes modifying, contain synthetic dione compounds thioesters and be used for initial condensation product, and need not compete with the dione compounds of natural generation.Therefore, this recombinant host can provide a kind of synthetic dione compounds thioesters to be used for mixing the polyketide that obtains.This dione compounds is mixed the polyketide that obtains, obtain having the substituent polyketide that to select as required at 13.U.S.'s series application number 60/117,384 that the preferred method for preparing this synthetic property polyketide thioesters is submitted to 27 days January in 1999 in application and submitted on January 27th, 2000 09/492,733 in list, be incorporated herein for your guidance.

PCT application WO97/02358 that published on January 28th, 1997 and the WO99/03986 that published on January 28th, 1999 have described the recombinant forms of the 6-dEBPKS of ketone synthetic enzyme (KS) structural domain that contains inactivation in first assembly (KS1), the suitable organism that contains this PKS expression system with modified is incorporated herein for your guidance.

The U.S. Provisional Application of submitting on May 6th, 1998 number 09/073,538, the U.S. Provisional Application of submitting on April 16th, 1999 number 60/429,09/429 of submission on October 28th, 349 and 1999,349 disclose other provides other substituent operations on the macrolide ring, be incorporated herein for your guidance.

Then as needing, the PKS that separates from the organism of recombinant modified and be purified into modification expresses the polyketide that obtains, it is fed to red saccharopolyspora (Saccharopolyspora erythraea), and it has rhetorical function behind the polyketide, comprises glycosylation.Other modifications are included in 6 and/or 12 hydroxylations.Separate the modification erythromycin that obtains then and carry out chemically modified, obtain compound of the present invention.WO98/09978 and U.S. Patent number 5,750,510 have been described the synthetic method that these modifications are provided, and are incorporated herein for your guidance.

Fig. 1 has shown the universal method of synthetic compound of the present invention.

The anti-infective compounds that obtains all has activity for one group of representative microorganism in vitro and in vivo.Therefore, compounds show of the present invention abundant diversity on the specificity, cover required antibiotic activity spectrum.

In order to be used for the treatment of infectious diseases, compound of the present invention is mixed with suitable composition, it will comprise typical vehicle, if compound is a salt, also comprise acceptable counterion on the pharmacology; As needs, also comprise other additives, as antioxidant, buffer reagent etc., and be applied to the animal or human.Be applicable to the similar of the preparation type of these compounds and general macrolide antibiotic.Can be at for example Remington ' s Pharmaceutical Sciences, Mack Publishing Co. finds these preparations in the latest edition.The approach of available any needs (comprising injection, orally administering, transdermal administration, transmucosal administration or any combination) administered compound.As needs, compound of the present invention also can be used with other activeconstituentss.

Compound of the present invention has formula (1)-(3) that propose above, and any stereoisomeric forms in any ratio of these shown compounds.The concrete steric isomer of describing obtains from above-mentioned synthetic preferred method, and illustration in the text; Yet, by revising the expression system of PKS, or by changing the chirality of dione compounds, or transform by synthetic chemistry, also can prepare other isomer.Also can in substituting group, there be additional chiral centre, as R _dAnd R _fThese isomer can be used as mixture and use, or can separate and utilize each isomer as known in the art like that.

The character of the compound of formula (1)-(3) is by substituent R _a-R _f, Y and Z limit.These substituent preferences have hereinafter been listed.They contain the group of following qualification:

" halogen " comprises fluorine, chlorine, bromine and iodine, most preferably is fluorine.

" alkyl " refers to saturated straight chain, side chain or cyclic hydrocarbon radical, contains the carbon of given number, and can contain one or more suitable heteroatomss; Similarly, alkenyl and alkynyl refer to straight or branched or ring-type hydro carbons substituting group, contain one or more pairs of keys or one or more triple bond respectively, and contain one or more suitable heteroatomss.

" aryl " refers to aromatic substituent, may contain one or more heteroatomss, as phenyl, naphthyl, quinolyl or phenanthryl.

" aralkyl ", " aromatic yl alkenyl " or " aromatic yl polysulfide yl " refer to substituting group, and wherein aryl is connected with substituted group by alkyl, alkenyl or alkynyl bonding respectively.Carbon number in aralkyl, aromatic yl alkenyl or aromatic yl polysulfide yl also is defined.

" amido aralkyl ", " amido aromatic yl alkenyl " or " amido aromatic yl polysulfide yl " refer to substituting group, and wherein aryl is connected with substituted group with alkyl, alkenyl or alkynyl bonding by amido respectively.Carbon number in amido aralkyl, amido aromatic yl alkenyl or amido aromatic yl polysulfide yl also is defined.

Therefore, comprise " assorted alkyl ", " heterochain thiazolinyl ", " assorted alkynyl ", " heteroaryl ", " heteroaralkyl " etc. in the substituting group that this paper limits.Suitable heteroatoms comprises N, O and S.

All aforementioned substituting groups can be not to be substituted or further to be replaced.Typical case substituting group comprise R ,-OR ,-SR ,-NR ₂,-COR ,-COOR ,-CONR ₂,-OOCR ,-NRCOR ,-OCONR ₂,-CN ,-CF ₃,-NO ₂,-SOR ,-SO ₂R, halogen, wherein each R is respectively H or alkyl, alkenyl, alkynyl, aryl, aralkyl or the assorted form of these groups as defined above.In addition, alkyl, alkenyl and alkynyl can replace by aryl or heteroaryl, they also can further be replaced itself.Aryl and heteroaryl also can be by alkyl, alkenyl or alkynyl or other aryl or heteroaryl replacements.

" deutero-oxime " is formula=N-O-R, and wherein R is not H, and is defined as above.

" blocking group " of hydroxyl comprises acyl group, silyl etc.At Greene, T.W. etc., ProtectGroups in Organic Synthesis, second edition, John Wiley ﹠amp; Sons, Inc. has described suitable substituents in (1991), is incorporated herein for your guidance.

The present invention includes the preferred embodiment that is defined as above.R _dPreferably butyl, amyl group, methoxy ethoxy methyl, isobutyl-, methylcyclohexyl, phenyl, benzyl, ethylphenyl, 3-(benzyloxy) propyl group, 2-(pyrimidine-2-base sulfo-) ethyl, propyl group, fluoro ethyl, chloroethyl, vinyl, 3-butenyl or azido-ethyl, more preferably propyl group, fluoro ethyl, chloroethyl, vinyl, 3-butenyl or azido-ethyl.The U.S. Patent number 60/117 that on January 27th, 1999 submitted to, the U.S. Patent number 09/492 that on January 27th, 384 and 2000 submitted to, 733 (all being incorporated herein for your guidance) have been described various few ketone compound thioesters, and preferred dione compounds thioesters can mix in the C-13 position.Dione compounds thioesters described in the literary composition can be mixed compound of the present invention, and the preferred R of definite C-13 position _dGroup.

In another preference, R _fBe H or rudimentary C1-C3 alkyl, more preferably methyl.R _fAlso preferred aryl groups alkenyl or aromatic yl polysulfide yl are as 3-arylprop-2-thiazolinyl or 3-aryl Propargyl.Preferred aryl groups is 3-quinolyl, 4-quinolyl, 5-quinolyl, phenyl, 4-fluorophenyl, 4-chloro-phenyl-, 4-p-methoxy-phenyl, 6-quinolyl, 6-quinoxalinyl, 6-amino-3-quinolyl or 4-isoquinolyl in preferred aryl groups alkenyl or aromatic yl polysulfide yl embodiment.

Synthesizing of The compounds of this invention

As mentioned above, preferably by suitable dione compounds being fed the host cell of giving modified microorganism (containing the 6-dEB PKS expression system of having rejected KS1) or methyl being provided in the C-13 position, then the polyketide that obtains is fed and (change to the red saccharopolyspora recombinant bacterial strain, do not produce 6-dEB), preparation is used for the microbiotic parent material of the present invention of any further chemosynthesis.Can prepare can hydroxylation 6-and the 12-position or the bacterial strain of hydroxylation 12-position only.In the later case ,-H replaced-OR _fIn addition, can prepare the only bacterial strain of hydroxylation 6-position.Pass through homologous recombination, the transformant that obtains now can not produce the 6-dEB as substrate polyketide competition thing, but produce 6-position and 12-position hydroxylation, 3-position and the glycosylation of 5-position of the modification polyketide of polyketide transformant generation streptomycete or other.If macrolide only has 12-position hydroxylation, and does not need 6-position hydroxylation (OR _fReplaced by H), can make up the red saccharopolyspora bacterial strain by the eryF '-hydroxylase gene that interrupts among the bacterial strain K40-67.In addition, can make the eryK gene lose function, can be easy to generate the embodiment of compound (1)-(3), wherein R _aBe H.

The glycosylation that produces erythromycin generates and diglycosyl form like the naturally occurring erythromycin series.If the compound of formula (3) will prepare from initial product, the hydroxyl of the cladinose ring that needs protection so (with 3 bindings), macrolide is substituent to be modified subsequently to be suitable for.

The erythromycin that the present invention modifies also can contain-OH at 6 except the C-13 position is modified, unless as above-mentioned OR _fReplaced by H.(wherein 6 is OR for the compound of structure formula (1), (2) and (3) _f), the compound of the formula (3) with blocking group is provided, form R _cAnd R _eAn embodiment.With suitable these protections of protection reagent is effectively, realizes in non-proton solution as diacetyl oxide, benzoyl oxide, chloroformic acid benzyl ester, hexamethyldisilazane or trialkylsilkl chlorine.Suitable aprotic solvent comprises methylene dichloride, chloroform, tetrahydrofuran (THF), N-Methyl pyrrolidone, methyl-sulphoxide (DMSO), dimethyl formamide (DMF) etc.Usable mixtures also.The protection of two sugared hydroxyls can simultaneously or successively be carried out in the formula (3).

" the hydroxyl, also must protect the ketone group of 9 on macrolide ring except protecting 2 ' and 4 of two glucosyl residues.Usually this realizes by ketone group being changed into the deutero-oxime.The particularly preferred example of R comprises not alkyl (1-12C), replacement or unsubstituted aryl (6-10C), alkyl (1-12C), replacement or the unsubstituted heteroaryl (6-10C), alkyl (1-12C) and the assorted alkyl that replace or replace (formula CR ' for example among formula=NOR ₂The substituting group of OR, wherein each R ' except above-mentioned R distinguishes the such of illustration, can form cycloalkyl ring (3-12C) altogether with other).Preferred deutero-oxime is formula=NOR, and wherein R is the isopropoxy cyclohexyl.

9-ketone group and 2 ' and 4 " after hydroxyl is protected, can be by in the presence of alkali, reacting with alkylating reagent, the 6-hydroxyl in the alkylation precursor becomes the compound of formula (3).Alkylating reagent comprises alkyl halide and sulphonate.For example, alkylating reagent can comprise toluenesulphonic acids methyl esters, 2-fluoro ethyl bromine, cinnamyl bromine, crotyl bromide, allyl bromide 98, propargyl bromide etc.In the presence of alkali (as potassium hydroxide, sodium hydride, potassium isopropoxide, potassium tert.-butoxide) and aprotic solvent, carry out alkylation.

Particularly preferred R _fBe methyl, allyl group and ethyl.

In case finished the alkylation of 6-hydroxyl, saccharide residue and macrolide ring can go protection.Glucosides part go protection as Green, T.W. etc., Protect Groups in Organic Synthesis, following.Similarly condition causes the deutero-oxime is changed into=NOH.If the formation of deutero-oxime does not take place simultaneously with going protection, can carry out the conversion of oxime separately.

Can take off oxime then and change into ketone group with standard method known in the art.Take off the oxime agent and comprise inorganic oxysulfide, as sodium bisulfite, sodium pyrosulfate, Sulfothiorine etc.At this point, can use protic solvent, as pure and mild their mixture of water, methyl alcohol, ethanol, Virahol, trimethyl silane.In general, in the presence of organic acid, carry out de-oxime reaction.

In the process this point or after, the compound of formula (3) can further operate in the group of introducing on the 6-hydroxyl after being converted to the compound of the formula (1) that hereinafter further describes or (2).It (is O-CH that initial replacement can provide the 6-O-allyl group easily ₂CH=CH ₂), it can obtain 6-O propylated compound, or handle with perosmic anhydride further by the reduction derivatize, obtains 2,3-dihydroxypropyl compound, its further esterification on each Sauerstoffatom.Also available metachloroperbenzoic acid is oxidation O-allyl deriv in aprotic solvent, obtains epoxy compounds, and its available amine or contain the heteroaryl compound open loop of N obtains containing the compound of the side chain of N, or can oxidation under the Wacker condition, obtains substituting group O-CH ₂-C (O)-CH ₃, or but ozonize obtains aldehyde.Aldehyde can be changed into oxime then, or with the reaction of suitable amine, and in the presence of the borohydride reductive agent, reduce, obtain amine.Also can oxime be changed into nitrile by in aprotic solvent, reacting with dehydrated reagent.Also can make O-allyl deriv and aryl halide in Heck condition (Pd (II) or Pd (O), phosphine and amine or mineral alkali) reaction down, obtain 3-arylprop-2-thiazolinyl derivative.Use hydrogen and this derivative of palladium carbon reduction then, obtain 3-arylpropyl derivative.If initial substituent R _fBe the 2-propine, available similar reaction changes side chain, comprises arylation.

For the compound of formula (3) being transformed the compound of an accepted way of doing sth (1), by at first removing the cladinose group, with weakly acidic aqueous solution or go Glycosylase to handle the compound of formula (3).Suitable acid comprises the alcoholic solution of hydrochloric acid, sulfuric acid, Mono Chloro Acetic Acid, trifluoroacetic acid etc.Normally 0.5-24 hour reaction times, temperature is-10-35 ℃.In this reaction process, protect 2 ' group of the sugar that stays as mentioned above, going to go protection after the cladinose reaction then.Then the hydroxyl of the macrolide ring 3-position that obtains is oxidized to ketone with the Swern oxidation style of improvement.In the method, use oxygenant, as N-chloro-succinimide-dimethyl sulphide or carbide diamide-methyl-sulphoxide.Usually, in chlorinated solvent such as methylene dichloride, under-10-25 ℃ in preformed N-chloro-succinimide and dimethyl sulphide mixture the compound of adding formula (3).Stir after 0.5-4 hour, add tertiary amine (as triethylamine), produce corresponding ketone, remove 2 ' blocking group then.

For at 2 halogenation macrolides (with R _bBe transformed into halogen from H), with the compound of alkali and close electric halogenating agent (as perbromo-pyridine or N-fluorobenzene sulfonic acid) processing formula (1).After having prepared 3 ketone compounds, 2 of halogenations at any time are preferably after 11,12 rings form.

Can further operate the suitable substituent of C-13 position, as vinyl (vinyl), ethylidine (ethenyl), butenyl or azido-.For example, the derive amido acetate of The compounds of this invention of available aryl Acetyl Chloride 98Min. obtains the arylamino alkyl on the C-13 position.The C13 of preferred azido-derives and forms preceding generation at ketone compound.Deriving of ethylidine can take place before and after ketone compound forms.

For the compound of acquisition formula (2), handle the compound that obtains from formula (1) de-glycosylation reaction with dewatering agent (as carbonyl dimidazoles and alkali).

For the compound (wherein one of Z and Y are H, and the OH of another OH or protection is above-mentioned aminoderivative) of preparation formula (1)-(3), carbonyl or oxime or deutero-oxime use appropriate reductant (as sodium borohydride, Raney nickel/H ₂) reduction, or with sodium cyanoborohydride and the ammonification of amine reductibility.Also can obtain the amine of replacement by alkylation.

The compounds of this invention synthetic novel method also is provided.

One exemplary embodiment

Define the compound of formula (1), (2) and (3) with different substituents.Table 1 has shown the compound in the scope of the invention, and they are:

For formula (1), wherein R _aBe H or OH, R _bBe H, Cl or F, R _cBe H;

For formula (2), wherein R _aBe H or OH, R _cBe H; With

For formula (3), wherein R _aBe H or OH, R _cBe H, R _eBe H or group a, b, c or d;

Table 1
Table 1				??Rd	????Rf	????Y	????Z
??-CH ₃	????-CH ₂CH ₂-φ	????＝O		??Rd	????Rf	????Y	????Z
??-CH ₃	????-CH ₂CH ₂-φ	????＝O		??-CH＝CH ₂	????-CH ₂CH＝CH-φ	????＝O
??-CH ₂CH ₂CH ₃	????-CH ₂CH ₂NHCH ₃	????＝NOH		??-CH＝CH ₂	????-CH ₂CH＝CH-φ	????＝O
??-CH ₂CH ₂CH ₃	????-CH ₂CH ₂NHCH ₃	????＝NOH		??-CH ₃	????-CH ₂CHOHCH ₃	????＝NOCH ₂CH ₃
??-CH(CH ₃) ₂	????-CH ₂-φ	????H	?OH	??-CH ₃	????-CH ₂CHOHCH ₃	????＝NOCH ₂CH ₃
??-CH(CH ₃) ₂	????-CH ₂-φ	????H	?OH	??-CH ₃	????-CH ₂-CH＝CH ₂	????＝O
??-CH ₃	????-CH ₂-CH=CH-(3-quinolyl)	????＝O		??-CH ₃	????-CH ₂-CH＝CH ₂	????＝O
??-CH ₃	????-CH ₂-CH=CH-(3-quinolyl)	????＝O		??-CH ₃	????-CH ₂-CH ₂-CH ₂-(3-quinolyl)	????＝O

Table 1
Table 1			????Rd	Rf	????Y	????Z
????-CH ₃	-CH ₂-CH=CH-(2-methyl-6-quinolyl)	????＝O	????Rd	Rf	????Y	????Z
????-CH ₃	-CH ₂-CH=CH-(2-methyl-6-quinolyl)	????＝O	????-CH ₃	-CH ₂-CH=CH-(5-isoquinolyl)	????＝O
????-CH ₃	-CH ₂-CH=CH-(3-bromo-6-quinolyl)	????＝O	????-CH ₃	-CH ₂-CH=CH-(5-isoquinolyl)	????＝O
????-CH ₃	-CH ₂-CH=CH-(3-bromo-6-quinolyl)	????＝O	????-CH ₃	-CH ₂-C=CH-(6-methoxyl group-2-naphthyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-(2-phenyl vinyl)	????＝O	????-CH ₃	-CH ₂-C=CH-(6-methoxyl group-2-naphthyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-(2-phenyl vinyl)	????＝O	????-CH ₃	-CH ₂-C ≡ C-(3-quinolyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-naphthyl	????＝O	????-CH ₃	-CH ₂-C ≡ C-(3-quinolyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-naphthyl	????＝O	????-CH ₃	-CH ₂-C ≡ C-(6-methyl-2-naphthyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-(3-(2-furyl)-6-quinolyl)	????＝O	????-CH ₃	-CH ₂-C ≡ C-(6-methyl-2-naphthyl)	????＝O
????-CH ₃	-CH ₂-C ≡ C-(3-(2-furyl)-6-quinolyl)	????＝O	????-CH＝CH ₂	-CH ₃	????＝O
????-CH ₂OH	-CH ₂-C=CH-(4-fluorophenyl)	????＝O	????-CH＝CH ₂	-CH ₃	????＝O
????-CH ₂OH	-CH ₂-C=CH-(4-fluorophenyl)	????＝O	????-CH ₂OH	-CH ₂-C=CH-(3-quinolyl)	????＝O
????-CH ₂OH	-CH ₂-C=CH-(6-quinolyl)	????＝O	????-CH ₂OH	-CH ₂-C=CH-(3-quinolyl)	????＝O
????-CH ₂OH	-CH ₂-C=CH-(6-quinolyl)	????＝O	????-CH ₂OCH ₃	-CH ₂-C=CH-(3-pyridyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(3-quinolyl)	????＝O	????-CH ₂OCH ₃	-CH ₂-C=CH-(3-pyridyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(3-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-chloro-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(4-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-chloro-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(4-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-chloro-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-hydroxyl-3-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-chloro-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-hydroxyl-3-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-methoxyl group-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-aminocarboxyl-3-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-methoxyl group-3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-aminocarboxyl-3-quinolyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(3-(2-thiophenyl)-6-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-hydroxyl-2-naphthyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(3-(2-thiophenyl)-6-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C=CH-(6-hydroxyl-2-naphthyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(6-chloro-2-naphthyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(3-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(6-chloro-2-naphthyl)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(6-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂CH ₂NHCH ₂CH ₂-(2-chloro-phenyl-)	????＝O	????-CH ₂CH ₂CH ₃	-CH ₂-C ≡ C-(6-quinolyl)	????＝O
????-CH ₂CH ₂CH ₃	-CH ₂CH ₂NHCH ₂CH ₂-(2-chloro-phenyl-)	????＝O	????-CH ₃	-CH ₂CH ₂NH ₂	????＝O

Embodiment

The following examples are in order to illustrate rather than will to limit the present invention.

Visible compound number and name in illustrative flow process 1.

In these embodiments, in first general step of method, the fermentative preparation by reorganization streptomycete host cell 6-deoxidation erythronolide B (6-dEB) derivative compound.

Produce 15-methyl-6-deoxidation erythronolide B and 14, the fermentation of 15-dehydrogenation-6-deoxidation erythronolide B need be fed to fermentation cell synthetic dione compounds intermediate.The preparation of these synthetic dione compounds has been described in embodiment 1.These synthetic dione compounds are substrates of 6-deoxidation erythronolide B synthetic enzyme (DEBS), and this enzyme can not act on its natural substrate (propionyl CoA) owing to the sudden change in the ketone synthetase structure domain of the assembly 1 of DEBS.Prepared this reorganization DEBS with the plasmid pJRJ2 among streptomyces coelicolor (Streptomyces coelicolor) CH999.U.S. Patent number 5,672 has been described streptomyces coelicolor CH999 in 491, is incorporated herein for your guidance.The derivative of streptomyces coelicolor CH999 has been described in the Application No. 09/181,833 (being incorporated herein for your guidance), streptomyces coelicolor K39-02, it contains the ptpA gene through genetic modification, also can be used for this purpose.

Plasmid pJRJ2 coding eryAI, eryAII and eryAIII gene; EryAI gene contained in the plasmid contains the KS1 null mutation.The KS1 null mutation prevents the formation of the 6-deoxidation erythronolide B that wild type gene produces, unless the external source substrate is provided.The Application No. 08/675,817 of PCT publication number submission on July 5th, 99/03986 and 97/02358 and 1996; Describe plasmid pJRJ2 in 09/311,756 (all being incorporated herein for your guidance) that submit to 14,08/896,323 and 1999 on Mays of submitting on July 17th, 1997 and prepared the method for the erythromycin of novel 13-replacement with this plasmid.PCT number of patent application PCT/US00/02397 and the Application No. 09/492 that submit on January 27th, 2000 such as available contriver G.Ashley, 733 (the U.S. Patent applications 60/117 that all required submit in 1999,384 right of priority, being incorporated herein for your guidance) method that provides prepares the external source substrate, and comprises described compound.Also can use extragenic PKS gene except ery; Suitable gene comprises the KS1 null mutation that contains bamboo peach lactone and megalomycin PKS gene, as the Application No. of submitting on October 8th, 1,999 60/158,305, those that the PCT number of patent application US99/24478 (being incorporated herein for your guidance respectively) that submits to 22,09/428,517 and 1999 on the October of submitting on October 28th, 1999 describes.

Produce 14-and go the fermentation of first (nor)-6-deoxidation erythronolide B not need the feeding dione compounds, because recombinant host cell streptomyces coelicolor CH999/pCK7 has produced required compound.U.S. Patent number 5,672 has been described plasmid pCK7 in 491, and it comprises the DEBS gene.The derivative of also available plasmid pCK7, pKOS011-26.The host cell that contains pKOS011-26 and reorganization ptpA gene is streptomyces coelicolor 27-26/pKOS011-26.These host cells produce 6-deoxidation erythronolide B and 14-removes first-6-deoxidation erythronolids, because mix propionyl CoA and acetyl-CoA, they are as the substrate of DEBS.

The fermentation of streptomyces coelicolor CH999/pJRJ2 and streptomyces coelicolor CH999/pCK7 has been described among the embodiment 2.The 6-deoxidation erythronolids product that can obtain by screening and separating fermentation.

Then isolating product is added in the fermenting broth of red saccharopolyspora bacterial strain, produces other useful intermediate compounds of the present invention.The biosynthesizing of red saccharopolyspora bacterial strain catalysis 6-dEB derivative compound, and saccharide residue and 6-dEB derivative compound 3 and 5 combine.These bacterial strains also contain functional eryK gene product, and can be at 12 hydroxylation 6-dEB derivative compounds.The difference of bacterial strain is whether to produce functional eryF gene product.If produce, the compound of generation is 6 hydroxylations.If do not produce, just produce 6-deoxy erythromycin A derivative.In embodiment 3, describe these red saccharopolyspora fermentations, and from fermenting broth, separated the Erythromycin A derivative compound.

In the chemosynthesis of other intermediate compounds of the present invention, isolating product is used as intermediate then.For the erythromycin A derivant intermediate that contains the 6-hydroxyl; embodiment 4-6 has described this compound of alkylation; produce 6-O-alkyl intermediate of the present invention; embodiment 11 has described the method for allylation generation 6-O-allyl group intermediate; it can be shown in embodiment 15; " behind the hydroxyl protection and after protecting the 9-position as described in example 14 above, further derive 2 ' and 4.The flow process of these reactions as shown in Figure 3.

Embodiment 7-9 has described above-mentioned formula (3) compound and has transformed an accepted way of doing sth (1) compound and 10, the respective compound of 11-dehydrated form.This schematically shows in Fig. 4.

Embodiment 10 also listed preparation formula (3) 10, the method for 11-anhydro compounds, but OR wherein _fReplaced by H.This reaction process of these conversions as shown in Figure 5.

The compound of embodiment 11 can be converted into the compound of formula (1) or (2) shown in difference in embodiment 12 and 13.

Embodiment 16 has illustrated the halogenation of 2-position.

Embodiment 17 has illustrated that 15-azido-Erythromycin A changes into 15-amido erythromycin, as shown in Figure 6.

Embodiment 1

The preparation of dione compounds thioesters

This embodiment has described to be used to prepare and has fed the N-acetylcysteamine thioesters (NAcS) of giving reorganization streptomycete host cell, prepares 15-methyl and 14, the method for 15-dehydrogenation-6-deoxidation erythronolide B intermediate compound.Also described synthetic schemes described below in the novel number of patent application 09/492,733 of U.S. utility that the U.S. Provisional Patent Application of submitting on January 27th, 1999 is submitted to number on January 27th, 60/117,384 and 2000, both are incorporated herein for your guidance.

Therefore; make (4S)-N-[(2S; 3R)-2-methyl-3-hydroxyl caproyl]-4-benzyl-2-oxazolidone (preparation method D) and N-acetylcysteamine (preparation method B) reaction; prepared (2S; 3R)-and 2-methyl-3-hydroxycaproic ester NAcS (preparation method E), be used to prepare 15-methyl-6-deoxidation erythronolide B intermediate.And the N-acetylcysteamine is from N, S-diacetyl cysteamine (preparation method A) preparation.(4S)-N-[(2S, 3R)-2-methyl-3-hydroxyl caproyl]-4-benzyl-2-oxazolidone (preparation D) is with (4S)-N-propionyl-4-benzyl-2-oxazolidone (propionyl-NOx; Preparation method C) prepares.

Use similar approach; make (4S)-N-[(2S; 3R)-2-methyl-3-hydroxyl-4-pentanoyl]-4-benzyl-2-oxazolidone (preparation method F) and N-acetylcysteamine (preparation method B) reaction; prepared (2S; 3R)-2-methyl-3-hydroxyl-4-valerate NAcS (preparation method G); the latter is used to prepare 14,15-dehydrogenation-6-deoxidation erythronolide B intermediate.(4S)-N-[(2S, 3R)-2-methyl-3-hydroxyl-4-pentanoyl]-4-benzyl-2-oxazolidone (preparation D) is with (4S)-N-propionyl-4-benzyl-2-oxazolidone (propionyl-NOx; Preparation method C) prepares.

A.N, S-diacetyl cysteamine: in 1 liter of 3-neck round-bottomed flask (magnetic splash bar, 2 application of sample funnels and pH electrode are housed), add Mercaptamine (50.0 gram).Add entry (300 milliliters), the solution that stirs in cooled on ice.Add 8N NaOH, with pH regulator to 8.0.Diacetyl oxide (125 milliliters) is placed an application of sample funnel, 8N KOH (350 milliliters) is placed another application of sample funnel.In cysteamine solution, drip diacetyl oxide, add 8N KOH simultaneously, will react pH and maintain 8+/-1.After adding diacetyl oxide, with pH regulator to 7.0, stirred the mixture 75 minutes on ice with 1N HCl.Add solid NaCl to saturated, with the CH of 400 milliliters of portions ₂Cl ₂Extracted solution 4 times.Merge organic extract, use MgSO ₄Drying is filtered, and concentrating under reduced pressure obtains the light yellow oil of 68.9 grams (97% productive rate), places post crystallization for 4 ℃.

B.N-acetylcysteamine: N, S-diacetyl cysteamine (42.64 gram) places 2 liters of round-bottomed flasks, and the magnetic splash bar wherein is housed, and is dissolved in 1400 ml waters.Use N ₂Clean flask, mixture is chilled in the ice bath.Add potassium hydroxide (49.42 gram), under inert atmosphere, stirred the mixture 2 hours on ice.With pH regulator to 7, add solid NaCl with 6N HCl to saturated.With 500 milliliters of a CH ₂Cl ₂Extracting mixture 7 times.Merge organic extract, use MgSO ₄Drying is filtered, and concentrating under reduced pressure obtains the product of 30.2 grams (96% productive rate).This material of distillation before being about to use, fusing point 138-140 ℃/7 mmhg.

C. (4S)-N-propionyl-4-benzyl-2-oxazolidone (propionyl-NOx): 20 gram (4S)-4-benzyl-2-oxazolidones are housed in 1 liter of three neck round-bottomed flask of the drying that 500 milliliters of application of sample funnels and splash bar are housed, add a cover, lead to nitrogen with an anti-mouthful soft rubber ball.Add anhydrous THF (300 milliliters) with intubate, the solution that cooling obtains in-78 ℃ of dry ice/isopropanol are bathed.In the application of sample funnel, add 78 milliliters of n-Butyl Lithiums (hexane solution of 1.6M), be added in the reactant with slow liquid stream with intubate.Add distilled propionyl chloride (fusing point 77-79 ℃) 8.0 milliliters rapidly with syringe.Reaction stirred is 30 minutes in dry ice/isopropanol is bathed.

Take out reactant from ice bath, temperature is to＞0 ℃, and with 50 milliliters of saturated NH ₄The cancellation of the Cl aqueous solution.On rotatory evaporator, mixture is condensed into slurries.With the ether extracting slurries of 250 milliliters of portions 3 times.Merge organic extract, respectively with 50 milliliters of saturated NaHCO ₃MgSO is used in the aqueous solution and salt water washing ₄Drying is filtered and the concentrated yellow oil that obtains.Crystallization when this material is placed.Grind crystallization once with cold (20 ℃) hexane, obtain 21.0 gram (80% productive rate) white crystalline material, fusing point 41-43 ℃.

APCI-MS:m/z=234 (MH+), 178,117.1H-NMR (360MHz, CDCl ₃): δ 7.2-7.4 (5H, m); 4.67 (1H, m, H4); 4.14-4.22 (2H, m, H5); (3.30 1H, dd, J=3,13Hz, benzyl); (2.89-3.03 2H, m, H2 '); (2.77 1H, dd, J=9,13, benzyl); (1.20 3H, t, J=7Hz, H2 ').

D. (4S)-N-[(2S; 3R)-2-methyl-3-hydroxyl caproyl]-4-benzyl-2-oxazolidone: 19.84 gram N-propionyls ,-oxazolidones are housed in 2 liter of three neck round-bottomed flask of the drying that 500 milliliters of application of sample funnels, low-reading thermometer and splash bars are housed; add a cover logical nitrogen with anti-mouthful of soft rubber ball.Add anhydrous methylene chloride (100 milliliters) with intubate, the solution that cooling obtains in dry ice/isopropanol is bathed is to-65 ℃.In the application of sample funnel, add 100 milliliters of trifluoromethanesulfonic acid dibutyl boron (dichloromethane solution of 1.0M), it is added in the reactant with slow liquid stream with intubate.Drip triethylamine with syringe, temperature of reaction remains on-10 ℃.After this, reactant is moved in the ice bath, 0 ℃ was stirred 30 minutes.After this, reactant is moved in the dry ice/isopropanol bath, be cooled to-65 ℃.Add butyraldehyde (8.6 milliliters), reaction stirred 30 minutes rapidly with syringe.

Reactant transfer in ice bath, is added 100 milliliters of 1M aqueous phosphatics, pH7.0 (phosphate solution is made up of the phosphoric acid one and the potassium dihydrogen of equimolar amount) in the application of sample funnel.Add phosphate solution as early as possible, simultaneously temperature of reaction is remained on below 10 ℃.In the application of sample funnel, add 300 ml methanol then as early as possible, simultaneously temperature of reaction is maintained below 10 ℃.In the application of sample funnel, add 300 milliliters of 2: 1 methyl alcohol at last: 30% hydrogen peroxide.With its dropping and guarantee that temperature maintenance is below 10 ℃.After adding, reaction stirred 1 hour.Then, on rotatory evaporator, remove and desolvate, up to the residue slurries.With the ether extracting slurries of 500 milliliters of portions 4 times.Each uses MgSO then with organic extract that 250 milliliters of saturated sodium bicarbonate aqueous solutions and salt water washing merge ₄Dry extract filters and the concentrated light yellow oil that obtains.At SiO ₂Last with 2: 1 hexanes: ethyl acetate (product R _f=0.4) this material of chromatography obtains 22.0 gram (85% productive rate) colorless oil title compounds.

APCI-MS:m/z 306 (MH+); 1H-NMR (360MHz, CDCl ₃): δ 7.2-7.4 (5H, m, phenyl); 4.71 (1H, m, H4); 4.17-4.25 (2H, m, H5); (3.96 1H, m, H3 '); (3.77 1H, dq, J=2.5,7Hz, H2 '); (3.26 1H, dd, J=4,13Hz, benzyl); (2.79 1H, dd, J=9,13Hz, benzyl); (1.5-1.6 2H, m, H4 '); (1.3-1.5 2H, m, H5 '); 1.27 (3H, d, J=7Hz, 2 '-Me); (0.94 3H, t, J=7Hz, H6 ').

E:(2S, 3R)-2-methyl-3-hydroxycaproic ester N-acetylcysteamine thioesters: distillation N-acetylcysteamine under 130 ℃/7 mmhg obtains liquid colourless under the room temperature.Seal 1 liter of three neck round-bottomed flask of drying that 500 milliliters of application of sample funnels and splash bar are housed with anti-mouthful of soft rubber ball, logical nitrogen.Then with syringe at the flask 10.7 milliliters of N-acetylcysteamines of packing into, add 400 milliliters of anhydrous THF with intubate.With MeOH/ ice bath cooling mixture.Drip butyllithium (64 milliliters of 1.6M hexanes) with syringe, form white precipitate.Stir after 30 minutes, drip trimethyl aluminium (51 milliliters of 2.0M hexanes) with syringe.Add trimethyl aluminium afterreaction thing and become clarification, and stirred 30 minutes in addition.This time after date, with (the 4S)-N-[(2S of 20.5 grams (0.068 mole), 3R)-2-methyl-3-hydroxyl caproyl]-4-benzyl-2-oxazolidone places under one deck nitrogen, and is dissolved in 100 milliliters of anhydrous THF; This solution is transferred in the reactant with slow liquid stream with intubate then.The reaction mixture that obtains becomes yellow-green colour, and stirs 1 hour.When thin layer chromatography analysis is no longer seen parent material (about 1 hour), reaction is finished.

With capacity saturated oxalic acid processing reaction thing, obtain neutral reaction thing (about 90 milliliters) with the pH test paper.On rotatory evaporator, remove then and desolvate, obtain white slurries.With the ether extracting slurries of 250 milliliters of portions 6 times.Merge organic extract, and use the salt water washing, use MgSO ₄Drying is filtered and the concentrated light yellow oil that obtains.On SiO2 with 1: 1 hexane: EtOAc flash chromatography sulfur purification ester products, up to washing out 4-benzyl-2-oxazolidone.At this moment, solvent systems is switched to 100%EtOAc, obtain the pure component of dione compounds thioesters.Merge the product component, concentrate, obtain the title compound of 14.9 grams (89% productive rate).This compound is the propyl group dione compounds thioesters among the embodiment 2.

APCI-MS：m/z?248(MH+)；1H-NMR(360MHz，CDCl ₃)：δ5.8(br?s，1H)；3.94(dt，1H)，3.46(m，2H)，3.03(dt，2H)，2.71(dq，1H)，1.97(s，3H)，1.50(m，2H)，1.37(m，2H)，1.21(d，3H)，0.94(t，3H)

F. (4S)-N-[(2S; 3R)-2-methyl-3-hydroxyl-4-pentanoyl]-4-benzyl-2-oxazolidone: in 2 liter of three neck round-bottomed flask of the exsiccant that 500 milliliters of application of sample funnels, low-reading thermometer and splash bars are housed, add 20.0 gram Bing Xian oxazolidone A; with anti-mouthful of soft rubber ball sealing, feed nitrogen.Add anhydrous methylene chloride (100 milliliters), the solution that cooling obtains in methyl alcohol/ice bath is to-15 ℃.Add trifluoromethanesulfonic acid dibutyl boron (dichloromethane solutions of 100 milliliters of 1.0M) by the application of sample funnel with slow liquid stream, this speed remains below 3 ℃ with temperature of reaction.Drip diisopropyl ethyl amine (17.9 milliliters) with syringe, internal temperature is remained below 3 ℃ again.To react then with the dry ice/isopropanol bath and be cooled to-65 ℃.Added propenal through 5 minutes with syringe.Adding the back reaction stirred 30 minutes.

Then with reactant transfer in ice bath, in the application of sample funnel, add the phosphate solution (phosphoric acid solution is made of the monobasic and the binary phosphate of equimolar amount) of 120 milliliters of (0.1 mole) 1MpH7.0.Add phosphate solution as early as possible, keep temperature of reaction simultaneously and be lower than 10 ℃.In the application of sample funnel, add 400 ml methanol then as early as possible, simultaneously temperature of reaction is maintained below 10 ℃.At last, in the application of sample funnel, add 400 milliliters of 2: 1 methyl alcohol: 30% hydrogen peroxide (drip, maintain the temperature at below 10 ℃).Reaction stirred 1 hour.Except that desolvating, be left slurries with rotatory evaporator.With 500 milliliters of a ether extracting slurries 4 times.Merge organic extract, each uses MgSO then with 250 milliliters of saturated sodium bicarbonates and salt water washing ₄Drying is filtered and the concentrated light yellow oil that obtains.With hexane grinding causing crystallization.Add hexane recrystallization from ether, obtain the product of 13.67 grams (55% productive rate).

1H-NMR(360MHz，CDCl ₃)：δ7.2-7.4(m，5H)；5.86(ddd，1H)，5.35(dt，1H)，5.22(dt，1H)，4.71(m，1H)，4.51(m，1H)，4.21(m，2H)，3.89(dq，1H)，3.26(dd，1H)，2.80(dd，1H)，1.25(d，3H)。

G. (2S, 3R)-2-methyl-3-hydroxyl-4-amylene ester N-acetylcysteamine thioesters: distillation N-acetylcysteamine under 130 ℃/7 mmhg at room temperature obtains colourless liquid.With anti-mouthful of soft rubber ball sealing the drying of 500 milliliters of application of sample funnels and splash bar, 1 liter of three neck round-bottomed flask are housed, use nitrogen wash.Then in flask with the syringe 7.5 milliliters of N-acetylcysteamines of packing into, add 500 milliliters of anhydrous THF with intubate.Then with MeOH/ ice bath cooling reactant.Drip butyllithium (hexane solutions of 44 milliliters of 1.6M) with syringe.Along with the adding of n-Butyl Lithium, form white precipitate.Stir after 30 minutes, with syringe Dropwise 35 .5 milliliter (0.071 mole) trimethyl aluminium (2.0M hexane solution).Reactant becomes clarification, restir 30 minutes after adding trimethyl aluminium.Under nitrogen, add (4S)-N-[(2S from preparation method F, 3R)-2-methyl-3-hydroxyl-4-pentanoyl]-4-benzyl-2-oxazolidone (13.6 gram), be dissolved in 50 milliliters of anhydrous THF, then this solution is transferred in the reactant with intubate with slow liquid stream.The reaction mixture that obtains becomes yellow-green colour, stirs 1 hour.When can't see parent material (about 30 minutes), judge to react and finish with thin-layer chromatography.

Add capacity saturated oxalic acid (about 60 milliliters), obtain the neutral reaction thing with the test of pH test paper.Remove with rotatory evaporator then and desolvate, obtain white slurries.With a extracting slurries of 250 milliliters of ether 6 times.Merge organic extract, use the salt water washing, use MgSO ₄Drying is filtered, and concentrates and obtains light yellow oil.Then at SiO ₂Last with flash chromatography purifying thioesters.With 1: 1 hexane: ethyl acetate drip washing, Zhi washed out Dao oxazolidone.At this moment, leacheate changes 100%EtOAc into, washes out the pure products component.Merge component and concentrated, obtain the title compound product of 7.7 grams (71% productive rate).This product is called vinyl dione compounds thioesters in embodiment 2.

1H-NMR(360MHz，CDCl ₃)：δ5.82(ddd，1H)，5.78(br?s，1H)，5.32(dt，1H)，5.21(dt，1H)，4.47(m，1H)，3.45(m，2H)，3.04(m，2H)，2.81(dq，1H)，1.96(s，3H)，1.22(d，3H)。

Embodiment 2

The preparation of erythronolids

A.15-methyl-6-deoxidation erythronolide B (Compound P, R _a=H, R _d=propyl group)

In Application No. of submitting on July 17th, 1,997 08/896,323 and 08/675,817 (being incorporated herein for your guidance respectively) of submitting on July 5th, 1996 streptomyces coelicolor CH999/pJRJ2 has been described.The mutant form of plasmid pJRJ2 encoding D EBS, wherein the ketone synthetase structure domain of assembly 1 (KS1) is by mutagenesis be inactivated (KS1 °).Feed to give the streptomyces coelicolor bacterial strain embodiment 1 that contains this plasmid (2S, 3R)-2-methyl-3-hydroxycaproic acid-N-acetylcysteamine (preparation method E, propyl group dione compounds) produces 15-methyl-6-deoxidation erythronolide B.

Melt 1 milliliter of bottle CH999/pJRJ2 working cardial cell storehouse, vial content is added in 250 milliliters of 50 milliliters of inoculation culture liquid 1 in the flask that baffle plate arranged.Flask places the incubator/shaking table 48 ± 10 hours that maintains 30 ± 1 ℃ and 175 ± 25RPM.Then 50 milliliters of cultures are added in 2.8 liters of flasks that baffle plate arranged, contain 500 milliliters of inoculation culture liquid 1 in this flask.This flask was cultivated in incubator/shaking table 48 ± 10 hours under 30 ± 1 ℃ and 175 ± 25RPM.These 500 milliliters of culture five equilibriums 10 2.8 liters have in the baffle plate flask, are respectively contained 500 milliliters of inoculation culture liquid 1.All flasks are cultivated as described above then.

Produce nutrient solution 1 with 100 liters and sterilized 45 minutes, prepared 150 liters of fermentor tanks at 121 ℃.After the cultivation, all 10 liters of flasks are incorporated in 5 liters of sterile culture bottles aseptic being added in 150 liters of fermentor tanks.Fermentor tank is controlled at 30 ℃, adds 2.5N H ₂SO ₄Be controlled at pH6.5 with 2.5N NaOH,, keep dissolved oxygen 〉=80% air saturation with stirring velocity (500-700RPM), air velocity (10-50LPM) and/or back pressure control (0.1-0.4 crust).The 50% solution control foam that intermittently adds Antifoam B.

At 24 ± 5 hours, add (2S, 3R)-2-methyl-3-hydroxyl caproyl-N-ethanoyl cysteamine (propyl group dione compounds, the preparation method E of embodiment 1) to ultimate density is 1 grams per liter.The propyl group dione compounds is by being dissolved in methyl-sulphoxide preparation, filtration sterilization (0.2 micron, nylon leaching film) then with 1: 4 (dione compounds is than DMSO).(fermented product is collected in being created in the 7th and stopping of 15-methyl-6dEB) to 15-methyl-6-deoxidation erythronolide B.In Alpha Laval AS-26 whizzer, with 20, the centrifugal fermenting broth of 500g.Product is mainly in centrifugate; Abandon the centrifugal cell mass.

Also in 1000 liters of fermentor tanks (700 liters of working volumes), finished this process.Seeded process is identical with said process, and except 150 liters of fermentor tanks are equipped with inoculation culture liquid 1, and 1000 liters of fermentor tanks are equipped with production nutrient solution 1.Fermentor tank is controlled at 30 ℃, and pH6.5 (adds 2.5-5N H ₂SO ₄With 2.5-5N NaOH), with stirring velocity (140-205RPM), air velocity (100-200LPM) and/or back pressure control (0.2-0.5 crust), keep dissolved oxygen 〉=70% air saturation.The 50% solution control foam that adds Antifoam B on demand.At 24 ± 5 hours, in 1000 liters of fermentor tanks, add racemic 2-methyl-3-hydroxyl caproyl-N-propionyl cysteamine (300 gram).At centrifugal as mentioned above collection fermented product on the 4.6th.

The nutrient solution that uses in this method comprises following:

Inoculation culture liquid 1

Composition	Concentration
Composition	Concentration	????KNO ₃	2 grams per liters
The yeast extract	20 grams per liters	????KNO ₃	2 grams per liters
The yeast extract	20 grams per liters	????Hycase?SF	20 grams per liters

????FeSO ₄-7H ₂O	25 mg/litre
????FeSO ₄-7H ₂O	25 mg/litre	NaCl (12.5% liquid storage)	4 milliliters/liter
????MgSO ₄(12.5% liquid storage)	1 milliliter/liter	NaCl (12.5% liquid storage)	4 milliliters/liter
????MgSO ₄(12.5% liquid storage)	1 milliliter/liter	????MnSO ₄-H ₂O (0.5% liquid storage)	1 milliliter/liter
????ZnSO ₄-7H ₂O (1.0% liquid storage)	1 milliliter/liter	????MnSO ₄-H ₂O (0.5% liquid storage)	1 milliliter/liter
????ZnSO ₄-7H ₂O (1.0% liquid storage)	1 milliliter/liter	????CaCl ₂-2H ₂O (2.0% liquid storage)	1 milliliter/liter

With 121 ℃ of sterilizations of autoclave 60 minutes.

Add the sterilization back:

1) the 100%DMSO solution of the thiostrepton of every liter of 1 milliliter of 50 mg/ml, sterile filtration.

2) 1 milliliter every liter 100%Antifoam B silicon emulsion (J.T.Baker), autoclaving.

3) glucose of 40 milliliter of 500 grams per liter, sterile filtration.

Produce nutrient solution 1

Composition	Grams per liter
Composition	Grams per liter	W-Gum	????45
Corn leaching solution	????10	W-Gum	????45
Corn leaching solution	????10	The cereuisiae fermentum of drying, deactivation	????10
????CaCO ₃	????1	The cereuisiae fermentum of drying, deactivation	????10

In fermentor tank, sterilized 45 minutes for 121 ℃.

Producing nutrient solution 1 sterilization back adds:

After centrifugal, filter centrifugate.Filtrate (about 700 liters) is by containing the Amicon Moduline post (20 * 350 centimetres) of 20 liters of HP20 resins (Mitsubishi).Flow velocity is 4 liters/minute when application of sample, and pressure drop is less than 8psi.Behind the application of sample, use 20 premium on currency, use 40 liter of 30% methanol wash resin then.With 100% methanol-eluted fractions 15-methyl-6dEB.Collect 4 12 liters of components, wherein component 2,3 and 4 contains whole detectable 15-methyl-6dEB.15-methyl-6dEB product with the dilution of 36.7 premium on currency merges obtains 75 liters of settled solutions.This solution directly is added on 5 liters of Amicon Vantage posts that contain HP20SS resin (Mitsubishi).With 1 liter of/minute upper prop.With 20 liter of 65% methyl alcohol, 20 liter of 70% methyl alcohol, 20 liter of 80% methyl alcohol and final 20 liter of 100% methanol-eluted fractions post.Collect whole 16 * 5 liters of components.Merge 80% component and last 70% component (25 liters) and be evaporated to dried.The residuum that obtains is dissolved in 1 liter of 100% methyl alcohol, filters, evaporation, 40 ℃ of dryings in vacuum oven.This process obtains 33 gram solid products, contains 15-methyl-6dEB of 93%.

B.14,15-dehydrogenation-6-deoxidation erythronolide B (Compound P, R _a=H, R _d=allyl group)

When the preparation method A according to above-mentioned generation 15-methyl-6-deoxidation erythronolide B prepares, contain this plasmid, and feed (2S to embodiment 1,3R)-the streptomyces coelicolor bacterial strain of 2-methyl-3-hydroxyl-4-valeric acid NAc cysteamine thioesters (preparation method G), produce 14,15-dehydrogenation-6-deoxidation erythronolide B.

C.14-remove first-6-deoxidation erythronolide B (Compound P, R _a=H, R _d=methyl):

Similarly, when the method for describing according to embodiment 2A prepares,, produced 14-without the dione compounds thioesters and removed first-6-deoxidation erythronolide B with streptomyces coelicolor CH999/pCK7 host.

Embodiment 3

The preparation of erythromycin

The 6-dEB derivative compound that preparation method A-C among the embodiment 2 is produced changes into erythromycin derivatives with the recombinant bacterial strain of red saccharopolyspora.In order to produce the erythromycin that has 6 and 12 hydroxyls simultaneously, the red saccharopolyspora bacterial strain of use is K40-67 or K39-14V.Produce the red saccharopolyspora bacterial strain of high-level Erythromycin A with pWHM3-deutero-plasmid (the eryA1 sequence that contains sudden change, the KS1 structural domain of coding inactivation) conversion energy, set up this bacterial strain.By homologous recombination, make the transformant that obtains to produce 6-deoxidation erythronolide B.Therefore feed and can not run into the hydroxylated competition in 6-position to the dEB1 analogue.In order to produce the erythromycin derivatives that only has the 12-hydroxyl, used red saccharopolyspora is K39-07.Made up this bacterial strain by interrupting the eryF '-hydroxylase gene from bacterial strain K40-67; This has destroyed the ability of hydroxylation 6-position analogue.These two kinds of bacterial strains of fermentation as described below under essentially identical condition.

15-methyl-Erythromycin A: produced 15-methyl-Erythromycin A: melt the K39-14V working cardial cell storehouse of 1 milliliter of bottle, the bottle content is added in 250 milliliters of 50 milliliters of inoculation culture liquid 2 in the flask that baffle plate arranged according to following method.Flask places the incubator/shaking table 48 ± 10 hours that maintains 34 ± 1 ℃ and 175 ± 25RPM.Then 50 milliliters of cultures are added in 2.8 liters of flasks that baffle plate arranged, contain 500 milliliters of inoculation culture liquid 2 in this flask.This flask was cultivated in incubator/shaking table 48 ± 10 hours under 34 ± 1 ℃ and 175 ± 25RPM.500 milliliters of culture five equilibriums 10 2.8 liters have in the baffle plate flask, are respectively contained 500 milliliters of inoculation culture liquid 2 in the flask.All flasks are cultivated as described above then.

Produce nutrient solution 2 with 100 liters and sterilized 45 minutes, prepared 150 liters of fermentor tanks at 121 ℃.After the cultivation, all 10 flasks are incorporated in 5 liters of sterile culture bottles, and aseptic being added in 150 liters of fermentor tanks.Fermentor tank is controlled at 34 ℃, and pH7.0 (adds 2.5N H ₂SO ₄With 2.5N NaOH), with stirring velocity (500-700RPM), air velocity (15-50LPM) and/or back pressure control (0.1-0.4 crust), keep dissolved oxygen 〉=80% air saturation.The 50% solution control foam that adds Antifoam B.

At 24 ± 5 hours, beginning added 15% dextrin (w/v) with 58-60 milliliter per hour.Constantly mix dextrin solution during the adding.At 24 ± 5 hours, 25 gram 15-methyl-6dEB (the preparation A among the embodiment 2) are added fermentor tank.Be dissolved in 400-600 milliliter 100% ethanol 25 gram 15-methyl-6dEB (the preparation A among the embodiment 2) and filtration (0.2 micron, nylon leaching film), prepared 15-methyl-6dEB.15-methyl-6dEB stops after 60 ± 10 hours to the conversion of 15-methyl-Erythromycin A, collects fermented product.In Alpha LavalAS-26 whizzer, with 20, the centrifugal fermenting broth of 500g.Product is mainly in centrifugate; Abandon the centrifugal cell mass.

The nutrient solution that uses in this method comprises following:

Inoculation culture liquid 2

Composition	Grams per liter
Composition	Grams per liter	W-Gum	????16.0
Corn dextrin	????10.0	W-Gum	????16.0
Corn dextrin	????10.0	Soyflour	????15.0
????CaCO ₃	????4.0	Soyflour	????15.0
????CaCO ₃	????4.0	Corn steep liquor	????5.0
Soybean oil	????6.0	Corn steep liquor	????5.0
Soybean oil	????6.0	????NaCl	????2.5
????(NH ₄) ₂SO ₄	????1.0	????NaCl	????2.5

With 121 ℃ of sterilizations of autoclave 60 minutes.

Add the sterilization back:

1 milliliter every liter 100%Antifoam B (J.T.Baker), autoclaving.

Produce nutrient solution 2

Composition	Grams per liter
Composition	Grams per liter	W-Gum	????17.5
Corn dextrin (3 type)	????16.0	W-Gum	????17.5
Corn dextrin (3 type)	????16.0	Soyflour	????16.5
????CaCO ₃	????4.0	Soyflour	????16.5
????CaCO ₃	????4.0	Corn steep liquor	????6.0

Soybean oil	????3.0
Soybean oil	????3.0	????NaCl	????3.5
????(NH ₄) ₂SO ₄	????1.0	????NaCl	????3.5

In fermentor tank, sterilized 45 minutes for 121 ℃.

The fermenting broth (127 liters) of centrifugal mistake that contains 34 gram target molecules is by being seated in 18.3 liters of HP20 sorbing agents on Amicon P350Moduline 2 chromatography columns.In 4 liters of/minute application of samples, find that pressure drop is less than 5psi.Behind the application of sample,, use 40 liter of 30% methanol wash resin then with 20 liters of deionized waters.Methanol-eluted fractions 15-methyl-Erythromycin A with 54 liter 100%.Product with Buchi rotatory evaporator (R-152) evaporation merging.Solid is dissolved in 100% minimum methyl alcohol, and filtration and evaporated filtrate are to doing.This obtains the material that 123 grams contain the 15-methyl-Erythromycin A of 30% weight.This 30% material with 1 liter of 40 ℃ of acetone extracting, 80 grams.Filter the acetone extract, dried filtrate on the internal surface of 20 liters of rotary evaporation flasks.With 9: 1 hexanes: acetone was 40 ℃ of extracting solids 3 times.Merge organic extract, be evaporated to dried, obtain 32 the gram solids, (68%) 15-methyl-Erythromycin A of enrichment.The merging product that the acetone/hexane extracting is obtained is dissolved in 1 liter of methyl alcohol (adding therein waits water gaging).Methanol solution is added to uses on 50% methanol wash and the equilibrated HP20SS chromatography column (Kontes).The size of post is 4.8 * 115 centimetres.Column load for 15-methyl-Erythromycin A is 11 grams per liters.Methanol aqueous solution washing column with 50% (0.8 liter) and 60% (8 liters).With 70% (8 liters), 80% (16 liters) and 85% (8 liters) methanol aqueous solution wash-out target molecule.Collect 1 liter of component.Merge component 11-29, evaporation is also dry in vacuum oven, obtains the product of 23 grams, 93% purity.

This material is as the parent material of chemical derivatization in the following example.Also produce following compounds: 14-with this method and removed clarithromycin A (R _d=Me); 14,15-dehydrogenation-Erythromycin A (R _d=allyl group); 14-removes first-6-deoxidation-Erythromycin A; 14,15-dehydrogenation-6-deoxidation-Erythromycin A; With 15-methyl-6-deoxidation-Erythromycin A.When being used to prepare 3-and going cladinose-3-oxo-derivative, erythromycin A derivant discord Erythromycin C derivative separately; But with the mixture of Erythromycin A and Erythromycin C compound as chemically derived parent material.

The method of extracting and these products of purifying is as follows:

Usually, add NaOH fermenting broth is transferred to pH8.0, add ethanol (0.1 liter/1 liter meat soup).Centrifugal clarification meat soup, with 2-4 milliliter/square centimeter-minute flow velocity be added on XAD-16 resin (Rohm and the Haas) post (1 kilogram of XAD/1 clarithromycin analogue).With 20% (v/v) aqueous ethanolic solutions of 2 column volumes washing application of sample resin, go out the erythromycin congener with acetone wash-out from the post, collect component with 1/2 column volume.With thin-layer chromatography (ethyl acetate: hexane 1: 1) and HPLC/MS identify the component contain the erythromycin analogue.

Merge the acetone component that contains the erythromycin analogue, volatile matter is removed in decompression.The aqueous mixture that obtains with the ethyl acetate extracting.Use saturated NaHCO ₃With salt brine solution washing ethyl acetate extract, with sodium sulfate or dried over mgso, filter, be evaporated to dried.Thick material is dissolved in methylene dichloride, is added on the silica gel short column, use methylene dichloride: (96: 4v/v) washing no longer is yellow up to elutant to methyl alcohol.Required material is with methylene dichloride: methyl alcohol: and triethylamine (94: 4: 2v/v) wash-out, collect component.Identify the component that contains erythromycin with thin-layer chromatography, collect and concentrating under reduced pressure.With ethylene dichloride/this material of hexane recrystallization.

The following universal method that illustrated:

(i) 14-goes clarithromycin: add 1 liter of ethanol in per 10 liters of fermenting broths.Centrifugal meat soup, supernatant liquor with the flow velocity of 100 ml/min by 0.6 liter of XAD (post is of a size of 6.5 centimetres of 17 cm x).Behind the application of sample, with 1.5 liter of 20% (v/v) aqueous ethanolic solution washing post.Use acetone wash-out desired substance then.Concentrating under reduced pressure contains the component of this material, up to removing volatile matter, with ethyl acetate extracting water residuum.With saturated sodium bicarbonate solution, salt water washing ethyl acetate layer, use dried over mgso, concentrating under reduced pressure obtains thick extract.

Thick material (0.6 gram) is dissolved in methylene dichloride, and gravity filters 3 centimetres of silicagel pad in the sand core funnel of 6 cm diameters.With 400 milliliters of methylene dichloride, use 400 milliliters of methylene dichloride then: methyl alcohol: triethylamine (90: 10: 2v/v) eluted material, collect component with per 40 milliliters.With thin-layer chromatography (ether: methyl alcohol: NH ₄OH90: 8: 2v/v, R _f～0.35 and methylene dichloride: methyl alcohol 95: 5v/v, R _f～0) identifies the component that contains erythromycin, concentrating under reduced pressure.This material of recrystallization from dichloromethane/hexane.

(ii) 15-methyl-erythromycin: 8 liters of ethanol are added in about 80 liters of fermenting broths.Centrifugal meat soup, supernatant liquor passes through 2.5 liters of XAD with 230 ml/min.Behind the application of sample, with the aqueous ethanolic solution washing post of 1 premium on currency and 5 liter 20% (v/v).Use acetone wash-out desired substance then.Concentrating under reduced pressure contains the component of this material, up to removing volatile matter, with ethyl acetate extracting water residuum.With saturated sodium bicarbonate solution, salt water washing ethyl acetate layer, obtain thick extract with dried over mgso and concentrating under reduced pressure.

Thick material (8.3 gram) is dissolved in methylene dichloride, and gravity filters 3 centimetres of silicagel pad in the sand core funnel of 9 cm diameters.With 200 milliliters of methylene dichloride, use 600 milliliters of methylene dichloride then: methyl alcohol (96: 4v/v), use 900 milliliters of methylene dichloride then: methyl alcohol: triethylamine (89: 9: 2v/v) eluted material, and collect 40 milliliters of components.With thin-layer chromatography (ether: methyl alcohol: NH ₄OH 90: 8: 2v/v, R _f~ 0.4, methylene dichloride: methyl alcohol 95: 5, R _f~ 0.05) identifies the component that contains erythromycin, concentrating under reduced pressure.This material carries out said process again, is fit to recrystallization up to it.

(iii) 14-removes first-6-deoxidation-erythromycin: respectively add 1 liter of ethanol in 2 10 liters of fermentations.Centrifugal meat soup merges supernatant liquor, amounts to about 22 liters.The meat soup that makes merging then with 170 ml/min flow velocitys by 1 liter of XAD (column dimension 23.5 cm x 6.5 centimetres (internal diameter)).Behind the application of sample, with 2 liter of 20% (v/v) aqueous ethanolic solution washing post.Use acetone wash-out desired substance then.Concentrating under reduced pressure contains the component of this material, up to removing volatile matter, with ethyl acetate extracting water residuum.With saturated sodium bicarbonate solution, salt water washing ethyl acetate layer, use dried over mgso, concentrating under reduced pressure obtains thick extract.

(iv) 15-methyl-6-deoxidation-erythromycin: in 3 fermentor tanks that respectively contain 10 liters of meat soups, respectively add 1 liter of ethanol.Centrifugal meat soup, supernatant liquor passes through 1.25 liters of XAD (6.5 centimetres of column dimension 40 cm x) with 130 ml/min flow velocitys.Use 3 liter of 20% (v/v) aqueous ethanolic solution washing post then.Use acetone wash-out desired substance then.Concentrating under reduced pressure contains the component of this material, up to removing volatile matter, with ethyl acetate extracting water residuum.With saturated sodium bicarbonate solution, salt water washing ethyl acetate layer, use dried over mgso, concentrating under reduced pressure obtains thick extract.

Thick material (2.8 gram) is dissolved in methylene dichloride, and gravity filters 3 centimetres of silicagel pad in the sand core funnel of 6 cm diameters.With 400 milliliters of methylene dichloride: methyl alcohol (96: 4v/v), use 400 milliliters of methylene dichloride then: methyl alcohol: triethylamine (89: 9: 2v/v) eluted material, and collect components with per 40 milliliters.With thin-layer chromatography (ether: methyl alcohol: NH ₄OH 90: 8: 2v/v, methylene dichloride: methyl alcohol 95: 5) identify the component that contains erythromycin, concentrating under reduced pressure.This material demand is further purified with silica gel column chromatography.

Embodiment 4

6-O-methyl isophthalic acid 4-removes clarithromycin A, i.e. formula (3) R wherein _a=OH, R _d=Me, R _f=Me, R _c=H, Z, Y=O's is synthetic

A.14-remove clarithromycin A 9-oxime: with 14-go Virahol (2 milliliters) solution of clarithromycin A (0.621 gram, 80% purity), azanol (0.5 milliliter of 50% aqueous solution) and acetate (0.2 milliliter) place 50 ℃ 22 hours.With chloroform/ethanol (3/2) extracting,, use MgSO with sodium bicarbonate, salt water washing ₄Dry.Filter also vacuum-evaporation and obtain white solid state crude product (0.65 gram), in next step transforms, directly use.

B.14-remove clarithromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime: go to clarithromycin A 9-oxime (0.65 gram) and 1 at above-mentioned thick 14-, add methylene dichloride (2 milliliters) solution of pyridine tosilate (PPTS) (0.333 gram) in methylene dichloride (2 milliliters) solution of 1-diisopropoxy-pimelinketone (0.95 milliliter).After stirring is spent the night, extracting (chloroform/ethanol 3: 2) mixture, washing (NaHCO ₃-H ₂O, salt solution), and dry (MgSO ₄).Behind filtration and the vacuum concentration, add toluene and Virahol and evaporate crude product repeatedly, obtain 0.74 gram product, in next step reaction, directly use.

C.2 ', 4 " two-O-trimethyl silyl-14-removes clarithromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime: remove clarithromycin A 9-[O-(1-isopropoxy cyclohexyl) at 14-] in methylene dichloride (6 milliliters) solution of oxime (0.74 gram), at 0 ℃ of methylene dichloride (2 milliliters) solution that adds trimethyl-silyl-imidazole (0.33 milliliter) and trimethylsilyl chloride (0.18 milliliter).After stirring in 5 minutes, add ethyl acetate, washing (NaHCO ₃-H ₂O, salt solution) and dry (MgSO ₄).Go up flash chromatography at silica gel (10: 1 hexanes: acetone, 1% triethylamine), obtain white solid state pure products (0.50 gram).Mass spectrum shows [M+H ⁺]=1020.

D.6-O-methyl-2 ', 4 " two-O-trimethyl silyl-14-removes clarithromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime: the diethyl ether solution with 0.3 milliliter of 2M monobromomethane handles 2 '; 4 "-two-O-trimethyl silyl-14-removes clarithromycin A9-[O-(1-isopropoxy cyclohexyl)] oxime (0.3 gram, 0.29 1: 1 methyl-sulphoxide/tetrahydrofuran (THF) (DMSO/THF) (1.4 milliliters) solution mmole) is cooled to 10 ℃.Added THF (0.6 milliliter) solution of 1M potassium tert.-butoxide and the mixture of DMSO (0.6 milliliter) through 6 hours with syringe pump.Use ethyl acetate diluting reaction thing then, use saturated NaHCO ₃, the salt water washing, use MgSO ₄Drying is filtered and vacuum-evaporation obtains white solid state crude product (0.29 gram).Mass spectrum shows [M+H ⁺]=1034.

E.6-O-methyl isophthalic acid 4-removes clarithromycin A 9-oxime: stir 6-O-methyl-2 ' at ambient temperature, 4 " two-O-trimethyl silyl-14-removes clarithromycin A 9-[O-(1-isopropoxy cyclohexyl)] mixture of oxime (0.29 gram), acetate (3.6 milliliters), acetonitrile (6 milliliters) and water (3 milliliters).Add the toluene evaporates mixture to doing, obtain white solid state crude product (0.24 gram), in next step without being further purified direct use.

F.6-O-methyl isophthalic acid 4-removes clarithromycin A: with 6-O-methyl isophthalic acid 4-go the mixture of clarithromycin A 9-oxime (0.24 gram), sodium bisulfite (0.45 gram, 85% purity), water (3 milliliters), ethanol (3 milliliters) and formic acid (0.07 milliliter) place 85 ℃ 8 hours.With 1N NaOH reactant is transferred to pH8, use the ethyl acetate extracting.With the organic extract of salt water washing, use MgSO ₄Drying is filtered and the concentrated white solid state crude product (0.2 gram) that obtains.Mass spectrum shows [M+H ⁺]=735.

Embodiment 5

6-O-methyl isophthalic acid 4,15-dehydrogenation Erythromycin A, i.e. formula (3) R wherein _a=OH, R _d=-CH=CH ₂, R _c=Me's is synthetic

A.14,15-dehydrogenation Erythromycin A 9-oxime:

Handle 14 with 1.97 milliliter of 50% aqueous hydroxylamine, 6 milliliters of 2-propyl alcohol suspensions of 15-dehydrogenation Erythromycin A (1.984 grams, 47% purity, 1.2 mmoles) stir up to dissolving.Add acetate (0.62 milliliter), mixture stirred 25 hours at 50 ℃.After the cooling, add saturated NaHCO at ambient temperature ₃, vacuum concentrated mixture is removed Virahol.With 250 milliliters of CHCl ₃The aqueous mixture that extracting obtains 3 times.Merge organic extract, use saturated NaHCO ₃, water and salt water washing, use MgSO then ₄Drying is filtered and the concentrated 0.92 gram product that obtains.

B.14,15-dehydrogenation Erythromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime:

To be dissolved in 6.2 milliliters of CH from the oxime (0.92 gram) of (A) ₂Cl ₂In, with 1,1-diisopropoxy-hexanaphthene (1.23 gram) and pyridine tosilate (PPTS) (0.464 gram) were handled 15 hours at ambient temperature.With 160 milliliters of CH ₂Cl ₂The diluted mixture thing is used saturated NaHCO then successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains brown slurries.At silica gel (toluene to 1: 1 toluene/acetone+1%Et ₃The gradient of N) goes up chromatography and obtain 0.998 gram product.

C.2 ', 4 " two (O-trimethyl silyls)-14,15-dehydrogenation Erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime

On ice, under inert atmosphere, cool off 14,15-dehydrogenation Erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] CH of oxime (998 milligrams, 9.96) ₂Cl ₂(11.25 milliliters) solution, and with the solution-treated of trimethylchlorosilane (0.24 milliliter) and 1-trimethyl-silyl-imidazole (0.44 milliliter).After 30 minutes,, use saturated NaHCO successively with 250 milliliters of ethyl acetate diluting reaction things ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains 1.002 gram products.

D.2 ', 4 " two (O-trimethyl silyl)-6-O-methyl isophthalic acids 4,15-dehydrogenation Erythromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime:

With 2 ', 4 " two (O-trimethyl silyls)-14; 15-dehydrogenation Erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] solution of 9.69 milliliters of 1: 1 tetrahydrofuran (THF)/methyl-sulphoxides of oxime (1.00 grams; 20.7 mmoles) is cooled to 10 ℃, and handles under inert atmosphere with the diethyl ether solution of 0.97 milliliter of 2.0M monobromomethane.The mixture that slowly adds the tetrahydrofuran (THF) (1.94 milliliters) of methyl-sulphoxide (1.94 milliliters) and 1.0M potassium tert.-butoxide.With thin-layer chromatography (silica gel, 10: 1 toluene/acetone) monitoring reaction, behind the alkali that adds 1.6 molar equivalents, judge and finish.With 200 milliliters of ethyl acetate and 70 milliliters of saturated NaHCO ₃The diluting reaction thing.Mixture is transferred in the separating funnel.With 850 milliliters of ethyl acetate and 280 milliliters of saturated NaHCO ₃Dilution, water and salt solution wash successively then.Use MgSO ₄Dry organic phase, filtration over celite and evaporation obtain the thick 6-O-methyl-2 ' of 21.2 grams, 4 " two-O-trimethyl silyl-14,15-dehydrogenation Erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime.Not purified use.

E.6-O-methyl isophthalic acid 4,15-dehydrogenation Erythromycin A 9-oxime:

With 5.3 milliliters of acetic acid treatment 6-O-methyl-2 ', 4 " two-O-trimethyl silyl-14,15-dehydrogenation Erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] solution of 9.8 milliliters of 2: 1 acetonitrile/water of oxime (1.0 gram), stirred 8 hours at ambient temperature.Vacuum concentrated mixture repeats to concentrate then after adding toluene, obtains the thick 6-O-methyl isophthalic acid 4 of 0.797 gram, 15-dehydrogenation Erythromycin A 9-oxime.

F.6-O-methyl isophthalic acid 4,15-dehydrogenation Erythromycin A:

With 6-O-methyl isophthalic acid 4, the solution of 7.5 milliliters of 1: 1 ethanol/waters of 15-dehydrogenation Erythromycin A 9-oxime (0.797 gram) and sodium bisulfite (85%, 1.02 gram) places under the inert atmosphere.Drip formic acid (0.186 milliliter), 80 ℃ stirred the mixture 3 hours.After being cooled to envrionment temperature, reactant is transferred to pH10, with 150 milliliters of ethyl acetate extractings 3 times with 6N NaOH.Merge organic extract, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains the 6-O-methyl isophthalic acid 4 that 0.68 gram is applicable to further conversion, 15-dehydrogenation Erythromycin A.

Embodiment 6

6-O-methyl-15-erythromycin A, i.e. formula (3) R wherein _a=OH, R _d=propyl group, R _f=Me's is synthetic

A.15-erythromycin A 9-oxime: handle 40 milliliters of 2-propyl alcohol suspensions of 15-erythromycin A (20.0 grams, 85% purity, 22.6 mmoles) with 20.5 milliliter of 50% aqueous hydroxylamine, stir up to dissolving.Add acetate (6.41 milliliters), mixture stirred 15 hours at 50 ℃.After being cooled to envrionment temperature, add saturated NaHCO ₃, vacuum concentrated mixture is removed Virahol.CHCl with 250 milliliters of portions ₃The aqueous mixture that extracting obtains 3 times.Merge organic extract, use saturated NaHCO ₃, water and salt water washing, use MgSO then ₄Drying is filtered and the concentrated 20.5 gram crude products that obtain.Obtain 94: 6 the E and the mixture of Z oxime, [M+H with the LC/MS analysis ⁺]=764.

B.15-erythromycin A-9-[O-(1-isopropoxy cyclohexyl)] oxime: above-mentioned oxime (20.5 gram) is dissolved in 55 milliliters of CH ₂Cl ₂In, with 1,1-diisopropoxy-hexanaphthene (27.3 milliliters) and pyridine tosilate (9.8 gram) were handled 15 hours at ambient temperature.With 160 milliliters of CH ₂Cl ₂The diluted mixture thing is used saturated NaHCO then successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains brown slurries.(gradient is hexane/acetone+1%Et from 2: 1 to 3: 2 at silica gel ₃N) go up chromatography and obtain 18.0 gram products.

C.2 ', 4 " two (O-trimethyl silyl)-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime: on ice; cooling 15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl) under inert atmosphere] 25 milliliters of CH of oxime (9.00 gram, 9.96 mmoles) ₂Cl ₂Solution, and with 8 milliliters of CH of trimethylchlorosilane (1.89 milliliters) and 1-trimethyl-silyl-imidazole (3.65 milliliters) ₂Cl ₂Solution-treated.After 30 minutes,, use saturated NaHCO successively with 250 milliliters of ethyl acetate diluting reaction things ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation.(gradient is from hexane to 10: 1 hexane/acetone+1%Et with silica gel column chromatography ₃N) purifying crude product obtains 7.8 gram products.

D.6-O-methyl-2 ', 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime: with 2 '; 4 "-two (O-trimethyl silyl)-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime (21.7 grams, 20.7 the solution of 41.4 milliliters of tetrahydrofuran (THF)s mmole) is cooled to 10 ℃, and handles under inert atmosphere with the diethyl ether solution of 41.4 milliliters of methyl-sulphoxides and 20.7 milliliters of 2.0M monobromomethanes.Tetrahydrofuran (THF) (41.4 milliliters) solution that adds methyl-sulphoxide (41.4 milliliters) and 1.0M potassium tert.-butoxide with about 20 milliliters of speed hourly.With thin-layer chromatography (silica gel, 10: 1 toluene/acetone) monitoring reaction, behind the alkali that adds 1.6 molar equivalents, judge and finish.With 200 milliliters of ethyl acetate and 70 milliliters of saturated NaHCO ₃The diluting reaction thing.Mixture is transferred in the separating funnel.With 850 milliliters of ethyl acetate and 280 milliliters of saturated NaHCO ₃Dilution, water and salt solution wash successively then.Use MgSO ₄Dry organic phase, filtration over celite and evaporation obtain the thick 6-O-methyl-2 ' of 21.2 grams, 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime.Not purified use.

E.6-O-methyl-15-erythromycin A 9-oxime: with 55 ml waters and 67 milliliters of acetic acid treatment 6-O-methyl-2 ', 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] 110 milliliters of acetonitrile solutions of oxime (21.2 gram), stirred 8 hours at ambient temperature.Vacuum concentrated mixture repeats to concentrate then after adding toluene, obtains 19.7 gram 6-O-methyl-15-erythromycin A 9-oximes.

F.6-O-methyl-15-erythromycin A: the solution of 280 milliliters of 1: 1 ethanol/waters of 6-O-methyl-15-erythromycin A 9-oxime (19.7 gram) and sodium bisulfite (85%, 23.1 restrains) is placed under the inert atmosphere.Drip formic acid (3.75 milliliters), 80 ℃ stirred the mixture 4.5 hours.After being cooled to envrionment temperature, use saturated NaHCO ₃The processing reaction thing is with 400 milliliters of a ethyl acetate extractings 3 times.Merge organic extract, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains 6-O-methyl-15-erythromycin A that 15.1 grams are applicable to further conversion.

Embodiment 7

5-O-(2 '-ethanoyl desoxy sugar amido)-10,11-dehydration-3-deoxidation-3-oxo-6-O-methyl isophthalic acid 4-remove the mould lactone A of methyl red (dehydrated form of formula (1), R _a=OH, R _d=Me, R _c=Ac, R _b=H) synthetic

A.5-O-desoxy sugar amido-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red: stir the mixture 3 hours that 6-O-methyl isophthalic acid 4-removes clarithromycin A (77 milligrams), 0.073 milliliter 12N HCl and water (2 milliliters) at ambient temperature.With 8N KOH mixture is transferred to pH8, use the ethyl acetate extracting.With the organic extract of salt water washing, use MgSO ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (3: 1/ hexanes: acetone, 1% triethylamine), obtain the pure products (42 milligrams) of white solid state.Mass spectrum discloses [M+H ⁺]=576.

B.5-O-(2 '-ethanoyl desoxy sugar amido)-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red: stir the mixture 18 hours that 5-O-desoxy sugar amido-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red (73 milligrams), salt of wormwood (20 milligrams), diacetyl oxide (14 microlitre) and acetone (1 milliliter) at ambient temperature.Add ethyl acetate, MgSO is used in water and salt water washing ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (3: 1/ hexanes: acetone, 1% triethylamine), obtain the pure products (71 milligrams) of white solid state.Mass spectrum discloses [M+H ⁺]=618.

C.5-O-(2 '-ethanoyl desoxy sugar amido)-3-deoxidation-3-oxo-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red (formula (1), R _a=OH, R _d=Me, R _f=Me, R _b=H, R _c=Ac): handle methylene dichloride (2 milliliters) solution that 5-O-(2 '-ethanoyl desoxy sugar amido)-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red (99 milligrams) and 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) hydrochloride (206 milligrams) with DMSO (0.21 milliliter), be cooled to 5 ℃.Methylene dichloride (2 milliliters) solution that in 4 hours, adds trifluoroacetic acid pyridine (208 milligrams) by the syringe pump.Add ethyl acetate then, use saturated NaHCO ₃, water, salt water washing, use MgSO ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (3: 1/ hexanes: acetone, 1% triethylamine), obtain the pure products (94 milligrams) of white solid state.Mass spectrum discloses [M+H ⁺]=616.

D.5-O-(2 '-ethanoyl desoxy sugar amido)-3-deoxidation-3-oxo-11-O-methylsulfonyl-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red: go in anhydrous pyridine (1 milliliter) solution of the mould lactone A of methyl red (93 milligrams) at 5-O-(2 '-ethanoyl desoxy sugar amido)-3-deoxidation-3-oxo-6-O-methyl isophthalic acid 4-, add methylsulfonyl chloride (0.057 milliliter) at 5 ℃.5 ℃ after 3 hours, the reactant temperature to envrionment temperature, was placed 15 hours again.With ethyl acetate diluted mixture thing, use saturated NaHCO ₃(2x), water (3x), salt water washing, use MgSO ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (2: 1/ hexanes: acetone, 1% triethylamine), obtain the pure products (72 milligrams) of white solid state.Mass spectrum discloses [M+H ⁺]=695.

E.5-O-(2 '-ethanoyl desoxy sugar amido)-10,11-dehydration-3-deoxidation-3-oxo-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red: handle acetone (1 milliliter) solution 18 hours that 5-O-(2 '-ethanoyl desoxy sugar amido)-3-deoxidation-3-oxo-11-O-methylsulfonyl-6-O-methyl isophthalic acid 4-removes the mould lactone A of methyl red (73 milligrams) at ambient temperature with diazabicylo undecylene (32 microlitre).With ethyl acetate diluted mixture thing, use saturated NaHCO ₃, water, salt water washing, use MgSO ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (2: 1/ hexanes: acetone, 1% triethylamine), obtain the pure products (50 milligrams) of white solid state.Mass spectrum discloses [M+H ⁺]=598. ¹³C-NMR(CDCl ₃，100MHz)：δ207.02，204.50，169.63，168.72，142.52，139.40，101.87，80.61，80.02，77.14，72.66，71.48，69.09，63.56，51.35，50.56，47.12，40.61，39.73，37.36，30.36，21.32，21.06，20.96，20.67，18.45，14.34，13.89，13.55，13.45。

Embodiment 8

2 '-O-benzoyl-6-O-methyl-3-goes cladinosyl group-3-oxo-10,11-dehydration-14,15-dehydrogenation Erythromycin A (dehydrated form of formula (1), R _a=OH, R _d=allyl group, R _f=Me, R _b=H, R _cSynthesizing=benzoyl)

A.2 '-and O-benzoyl-6-O-methyl isophthalic acid 4,15-dehydrogenation Erythromycin A

Stir 6-O-methyl isophthalic acid 4,3.6 milliliters of CH of 15-dehydrogenation Erythromycin A (668 milligrams), benzoyl oxide (385 milligrams) and triethylamine (0.25 milliliter) ₂Cl ₂Solution 2 days.Add saturated NaHCO ₃After, use CH ₂Cl ₂Extracting mixture 3 times.Merge organic extract, be evaporated to dried, with silica gel column chromatography purified product (90: 9: 1 toluene/acetone/Et ₃N), obtain 477 milligrams of products; LC-MS shows [M+H ⁺]=850.6.

B.2 '-and O benzoyl-6-O-methyl-4 ", 11-two (O-methylsulfonyl)-14,15-dehydrogenation Erythromycin A

Stir 2 '-O-benzoyl-6-O-methyl isophthalic acid 4,2.39 milliliters of pyridine solutions of 15-dehydrogenation Erythromycin A (549 milligrams) and methylsulfonyl chloride (0.50 milliliter) 24 hours are used CH then ₂Cl ₂With saturated NaHCO ₃Dilution.Use CH ₂Cl ₂Extracting mixture 3 times.Merge organic extract and be evaporated to dried, with silica gel column chromatography (90: 9: 1 toluene/acetone/Et ₃N) purified product obtains 530 milligrams of products.LC-MS shows [M+H ⁺]=1006.5.

C.2 '-O benzoyl-6-O-methyl-4 " O-methylsulfonyl-10,11-dehydration-14,15-dehydrogenation Erythromycin A

Stir 2 '-O-benzoyl-6-O-methyl-4 ", 11-two (O-methylsulfonyl) 14, the mixture of 0.195 milliliter of acetone of 15-dehydrogenation Erythromycin A (59 milligrams) and diazabicylo undecylene (0.018 milliliter) 24 hours, vacuum-drying then.With silica gel column chromatography (90: 9: 1 toluene/acetone/Et ₃N) purified product obtains 50 milligrams of products.LC-MS shows [M+H ⁺]=910.5.

D.2 '-O benzoyl-6-O-methyl-3-removes cladinosyl group-10,11-dehydration-14,15-dehydrogenation Erythromycin A

" O-methylsulfonyl-10,11-dewaters-14, the mixture of 15-dehydrogenation Erythromycin A (337 milligrams), 1.5 milliliters of acetonitriles and 6.9 milliliters of 3N HCl 22 hours to stir 2 '-O-benzoyl-6-O-methyl-4.Vacuum is removed acetonitrile, and adding NaOH transfers to 12 with the pH of water residuum, with 4 parts of CH ₂Cl ₂Extract product.The extract dry and evaporation merges.(gradient was from 96: 4 CH with silica gel column chromatography ₂Cl ₂/ MeOH to 95: 4: 1CH ₂Cl ₂/ MeOH/Et ₃N) purified product obtains 197 milligrams, [M+H ⁺]=674.4.

E.2 '-O-benzoyl-6-O-methyl-3-goes cladinosyl group-3-oxo-10,11-dehydration-14,15-dehydrogenation Erythromycin A

Stir 2 '-O-benzoyl-6-O-methyl-3-and remove cladinosyl group-10,11-dehydration-14,14.6 milliliters of CH of 15-dehydrogenation Erythromycin A (226 milligrams) and the high iodine alkane of Dess-Martin (periodinane) (427 milligrams) ₂Cl ₂(14.6 milliliters) 1 hour.Use CH ₂Cl ₂With saturated NaHCO ₃The diluted mixture thing.With 3 parts of CH ₂Cl ₂Extract product merges extract, dry and evaporation.Silica gel column chromatography (90: 9: 1 toluene/acetone/Et ₃N) obtain 168 milligrams of products, [M+H ⁺]=672.4. ¹³C-NMR (CDCl ₃, 100MHz): δ 206.78,203 (bromine), 168.19,165.08,141.36,139.58,132.74,131.51,130.46,129.79,128.25,120.18,102.09,80.79,80.40,78.70,72.52,71.91,69.19,63.76,51.10,50.54,47.08,40.73,39.87,37.77,31.23,22.13,20.98,18.52,14.28,14.15,13.55.

Embodiment 9

5-O-(2 '-ethanoyl desoxy sugar amido)-10,11-dehydration-3-deoxidation-3-oxo-6-O-methyl-15-methyl erythronolids (dehydrated form of formula (1), R _a=OH, R _d=propyl group, R _f=Me, R _b=H, R _c=Ac) synthetic

A.6-O-methyl-3-removes cladinosyl group-15-erythromycin A

Stirred 6-O-methyl-15-erythromycin A (15.1 gram) and 280 milliliters at ambient temperature 0.5N HCl3 hour.Add 6N NaOH with pH regulator to 9, collect the precipitation that obtains, wash with water and drying with vacuum filtration.With the ethyl acetate extracting filtrate of 400 milliliters of portions 3 times.Merge organic extract, use saturated NaHCO successively ₃, water and salt water washing, use MgSO then ₄Drying is filtered and evaporation, obtains another batch product.With the crude product that silica gel column chromatography merges, obtain the pure 6-O-methyl-3-of 9.35 grams and remove cladinosyl group-15-erythromycin A; ES-LC/MS shows [M+H ⁺]=605.

B.2 '-O-ethanoyl-6-O-methyl-3-removes cladinosyl group-15-erythromycin A

35 milliliters of ethyl acetate solutions that in 6-O-methyl-3-removes 40 milliliters of ethyl acetate solutions of cladinosyl group-15-erythromycin A (9.35 gram), add diacetyl oxide (2.92 milliliters).Stirred the mixture after adding 30 minutes, and concentrated then.Obtain 8.35 grams, 2 '-O-ethanoyl-6-O-methyl-3-with silica gel column chromatography (2: 1 hexane/acetone) and remove cladinosyl group-15-erythromycin A.ES-LC/MS shows [M+H ⁺]=647.

C.2 '-O-ethanoyl-6-O-methyl-3-removes cladinosyl group-3-oxo-15-erythromycin A

Go cladinosyl group-15-erythromycin A (8.3 gram) and 64 milliliters of methylene dichloride of 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride (16.51 gram) and the solution of 15.47 milliliters of methyl-sulphoxides to place under the inert atmosphere 2 '-O-ethanoyl-6-O-methyl-3-, in cooled on ice.Add 64 milliliters of dichloromethane solutions of trifluoroacetic acid pyridine (16.63 gram) with the speed that added with 4 hours, use the thin-layer chromatography monitoring reaction.After adding 73% solution, observe complete reaction, add 600 milliliters of ethyl acetate and 200 milliliters of saturated NaHCO then ₃The cancellation reaction.Collected organic layer is used saturated NaHCO ₃, water and salt solution washs successively, uses MgSO then ₄Drying is filtered and evaporation, obtains 8.4 gram crude products.With silica gel column chromatography (3: 1 hexane/acetone), obtain 6.75 grams, 2 '-O-ethanoyl-6-O-methyl-3-and remove cladinosyl group-3-oxo-15-erythromycin A.ES-LC/MS shows [M+H ⁺]=645.

D.2 '-O-ethanoyl-6-O-methyl-3-removes cladinosyl group-3-oxo-11-O-methylsulfonyl-15-erythromycin A

In removing 35 milliliters of pyridines of cladinosyl group-3-oxo-15-erythromycin A (6.73 gram), 2 '-O-ethanoyl-6-O-methyl-3-drips methylsulfonyl chloride (5.68 milliliters) at 0 ℃.Mixture is placed envrionment temperature, add 700 milliliters of ethyl acetate and 200 milliliters of saturated NaHCO ₃Cancellation.Collected organic layer is used saturated NaHCO successively ₃, water and salt water washing, use MgSO then ₄Drying is filtered and evaporation, obtains 8.2 gram crude products.Obtain 5.04 grams, 2 '-O-ethanoyl-6-O-methyl-3-with silica gel column chromatography (5: 2 hexane/acetone) and remove cladinosyl group-3-oxo-11-O-methylsulfonyl-15-erythromycin A.ES-LC/MS shows [M+H ⁺]=723.

E.2 '-O-ethanoyl-6-O-methyl-3-goes cladinosyl group-3-oxo-10,11-dehydration-15-erythromycin A

In removing 23 milliliters of acetone of cladinosyl group-3-oxo-11-O-methylsulfonyl-15-erythromycin A (5.03 gram), 2 '-O-ethanoyl-6-O-methyl-3-drips 1,8-diazabicylo [5.4.0] 11 carbon-7-alkene (5.22 milliliters).4.5 concentrate this solution after hour,, obtain 3.72 grams, 2 '-O-ethanoyl-6-O-methyl-3-and go cladinosyl group-3-oxo-10,11-dehydration-15-erythromycin A with silica gel column chromatography (5: 2 hexane/acetone) residuum.ES-LC/MS shows [M+H ⁺]=627.

Embodiment 10

5-O-(2 '-ethanoyl desoxy sugar amido)-10,11-dehydration-3,6-dideoxy-3-oxo-15-methyl erythronolids A (formula (1), dehydrated form, R _a=OH, R _d=propyl group, OR _fReplaced R by H _b=H, R _c=Ac)

In methylene dichloride (5 milliliters) solution of 6-deoxidation-15-erythromycin C (220 milligrams, 0.307 mmole), add salt of wormwood (50 milligrams) and diacetyl oxide (100 liters, 0.9 mmole), stirring at room reactant 16 hours.Filtering solution adds sodium hydroxide (1N, 25 milliliters) and salt solution (25 milliliters), uses ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo.In next step, use crude product (2 ' of parent material-acetylated form).

Crude product is dissolved in the pyridine (5 milliliters), adds methylsulfonyl chloride (70 liters, 0.9 mmole).-20 ℃ of reaction stirred 2 days are poured in sodium hydroxide (1N, 25 milliliters) and the salt solution (25 milliliters), with ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo." the two methylsulfonyl forms (190 milligrams, two step backs 68%) that with silica gel (toluene/acetone=3: 1,1% ammonium hydroxide) purifying residuum, obtain 11,4.

" two methylsulfonyl forms (190 milligrams, 0.21 mmole) are dissolved in acetone (7 milliliters), add DBU (63 liters, 0.42 mmole), and the stirring at room reactant spends the night with 11,4.Mixture is poured in sodium hydroxide (1N, 25 milliliters) and the salt solution (25 milliliters), uses ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo.In next step, use crude product (10 of 6-deoxidation-15-erythromycin, 11-dehydrogenation form).

In the crude product of above-mentioned steps, add hydrochloric acid (30 milliliters, 3N) and ethanol (2 milliliters), vigorous stirring mixture 6 hours.Add sodium hydroxide (5 milliliters 10N), are used ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo.In next step, use crude product (anhydrous form of formula (1) (but 3 be OH), wherein R _a=OH, R _d=propyl group, OR _fReplaced R by H _b=R _c=H).

In methylene dichloride (5 milliliters) solution of the crude product of above-mentioned steps, add diacetyl oxide (50 liters, 0.45 mmole) and salt of wormwood (100 milligrams), vigorous stirring mixture 9 hours.The filtering reaction thing, add sodium hydroxide (20 milliliters, 1N) and salt solution (25 milliliters), usefulness ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo.With silica gel (toluene/acetone=3: 1,1% ammonium hydroxide) purifying residuum, obtain 2 ' of parent material-acetylated form (110 milligrams, three step backs 89%).

The product (110 milligrams, 0.184 mmole) of above-mentioned steps is dissolved in methylene dichloride (10 milliliters), adds Dess-Martin reagent (220 milligrams, 0.53 mmole).Stirring at room reactant 45 minutes.With sodium hydroxide (20 milliliters, 1N) and salt solution (25 milliliters) cancellation reactant, with ethyl acetate extracting water layer 6 times.Organic layer with dried over sodium sulfate merges filters solvent removed in vacuo.(toluene/acetone, gradient=6: 1-3: 1,1% ammonium hydroxide) the chromatography purification residuum obtains the anhydrous form of the compound of formula (1), wherein R on silica gel _a=OH, R _d=propyl group, OR _fReplaced R by H _b=H, R _c=OAc (94 milligrams, 86%).

Embodiment 11

I. the compound of formula (3): R _a=OH, R _d=propyl group, R _f=allyl group

The allylation of step 1. intermediate microbiotic 6-OH: in cooled on ice 2 ', 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime (formula (I), (R _aBe OH, R _dIt is propyl group; 2 ' and 4 " protected by trimethyl silyl; protected by the isopropylcyclohexyl-oxime at C9=O)) 30 milliliters of tetrahydrofuran solutions of (7.8 gram, 7.44 mmoles), handle under inert atmosphere with 30 milliliters of methyl-sulphoxides and 2.58 milliliters of fresh distillatory allyl bromide 98s.Alkali/hour speed with 1.33 molar equivalents adds tetrahydrofuran (THF) (29.8 milliliters) mixture of methyl-sulphoxide (29.8 milliliters) and 1.0M potassium tert.-butoxide.With thin-layer chromatography (silica gel, 10: 1 toluene/acetone) monitoring reaction, add and judge that reaction finishes behind the alkali of 3.6 molar equivalents.With 700 milliliters of ethyl acetate diluting reaction things, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains the thick 6-O-allyl group-2 ' of 8.08 grams, 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime.Not purified use.

Step 2: with 21 ml waters and 24 milliliters of acetic acid treatment 6-O-allyl groups-2 ', 4 " two-O-trimethyl silyl-15-erythromycin A 9-[O-(1-isopropoxy cyclohexyl)] 42 milliliters of acetonitriles of oxime (8.08 gram), stirred 18 hours under the envrionment temperature.Enriched mixture behind the adding 2-propyl alcohol repeats behind the adding toluene, obtains 7.7 gram crude products.Silica gel column chromatography (2: 1 to 1: 1 hexane/acetone+1%Et of gradient ₃N) obtain 3.75 gram 6-O-allyl group-15-erythromycin A 9-oximes.

Step 3: the solution of 66 milliliters of 1: 1 ethanol/waters of 6-O-allyl group-15-erythromycin A 9-oxime (3.75 gram) and sodium bisulfite (85%, 5.37 restrains) is placed under the inert atmosphere.Drip formic acid (0.845 milliliter), 80 ℃ stirred the mixture 3.5 hours.After being cooled to envrionment temperature, reactant is adjusted to pH10, with the ethyl acetate extracting of 150 milliliters of portions 3 times with 6N NaOH.Merge organic extract, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains 3.42 gram 6-O-allyl group-15-erythromycin A, is applicable to further conversion.

II. the compound of formula (3): R _a=OH, R _d=Me, R _f=allyl group

Step 1: the allylation of intermediate microbiotic 6-OH: with 2 ', 4 " two-O-trimethyl silyl-14-removes clarithromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime (formula (I), (R _aBe OH, R _dIt is methyl; 2 ' and 4 " protected by trimethyl silyl; at C9=O by isopropoxy cyclohexyl oxime protection)) tetrahydrofuran (THF) (0.4 milliliter), DMSO (0.4 milliliter) and ether (0.04 milliliter) solution of (202 milligrams) is cooled to 10 ℃, handle under inert atmosphere with 0.035 milliliter of fresh distillatory allyl bromide 98.The mixture that adds the tetrahydrofuran (THF) (0.4 milliliter) of methyl-sulphoxide (0.4 milliliter) and 1.0M potassium tert.-butoxide with 0.22 milliliter of/hour speed.With thin-layer chromatography (silica gel, 5: 1 toluene/acetone) monitoring reaction.With ethyl acetate diluting reaction thing, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation obtains 222 milligrams of thick 6-O-allyl groups-2 ', 4 " two-O-trimethyl silyl-14-removes clarithromycin A 9-[O-(1-isopropoxy cyclohexyl)] oxime.Without being further purified use.

Step 2: with 2 ml waters and 2.4 milliliters of acetic acid treatment 6-O-allyl groups-2 ', 4 " two-O-trimethyl silyl-14-removes clarithromycin A 9-[O-(1-isopropoxy cyclohexyl)] 4 milliliters of acetonitrile solutions of oxime (222 milligrams), stirred 18 hours under the envrionment temperature.Enriched mixture behind the adding 2-propyl alcohol repeats behind the adding toluene, obtains 220 milligrams of thick 6-O-allyl group-14-and removes clarithromycin A 9-oxime.

Step 3: go the solution of 4 milliliters of 1: 1 ethanol/waters of clarithromycin A 9-oxime (220 milligrams) and sodium bisulfite (85%, 322 milligram) to place under the inert atmosphere 6-O-allyl group-14-.Drip formic acid (0.050 milliliter), 80 ℃ stirred the mixture 15 hours.After being cooled to envrionment temperature, reactant is adjusted to pH10, with the ethyl acetate extracting of 150 milliliters of portions 3 times with 6N NaOH.Merge organic extract, use saturated NaHCO successively ₃, water and salt water washing.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains 156 milligrams of 6-O-allyl group-14-and removes clarithromycin A, is applicable to further conversion.

Other embodiment: in a similar manner, from intermediate (R wherein _dBe butyl, benzyl, vinyl or 3-hydroxybutyl) prepared the compound of formula (3), wherein Y and Z are=O altogether, R _aBe OH, R _fIt is allyl group.

Embodiment 12

Transform an accepted way of doing sth (1)

Step 1: stir embodiment 11 at ambient temperature, the mixture of the compound for preparing among the II (77 milligrams, thick), 0.073 milliliter of 12N HCl and water (2 milliliters) 3 hours.With 8N KOH mixture is transferred to pH8, use the ethyl acetate extracting.With the organic extract of salt water washing, use MgSO ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (3: 1/ hexanes: acetone, 1% triethylamine), obtain white solid state pure products (42 milligrams).

Step 2:, stir the mixture 18 hours of above-claimed cpd (73 milligrams), salt of wormwood (20 milligrams), diacetyl oxide (14 microlitre) and acetone (1 milliliter) at ambient temperature in order to protect 2 ' OH.Add ethyl acetate, MgSO is used in water and salt water washing ₄Drying is filtered and evaporation.Go up the chromatography residuum at silica gel (3: 1/ hexanes: acetone, 1% triethylamine), obtain white solid state pure products (71 milligrams).

Step 3: the compound (99 milligrams) that obtains with DMSO (0.21 milliliter) treatment step 2 and methylene dichloride (2 milliliters) solution of 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDC) hydrochloride (206 milligrams), and be chilled to 5 ℃.With the syringe pump to add the solution of the methylene dichloride (2 milliliters) of trifluoroacetic acid pyridine (208 milligrams) in 4 hours.Add ethyl acetate then, use saturated NaHCO ₃, water, salt water washing, use MgSO ₄Drying is filtered and evaporation.With silica gel (3: 1/ hexanes: acetone, 1% triethylamine) chromatography residuum, obtain formula (1) pure compound (94 milligrams, R _aBe OH, R _cBe acetic ester, R _dBe CH ₃, R _fBe allyl group).

Step 4: in order to remove the protection of 2 ' OH, at room temperature 5 ml methanol solution of the compound (94 milligrams) that obtains of whipping step 3 are 24 hours.Under vacuum, remove and desolvate, obtain required formula (1) compound (R _aBe OH, R _cBe H, R _dBe CH ₃, R _fBe allyl group).

Other embodiment: use similar manner, prepared the compound of formula (1), wherein R _aBe OH, R _cBe H, R _fBe allyl group, R _dBe propyl group, butyl, benzyl, vinyl or 3-hydroxybutyl.

Embodiment 13

The preparation of the compound of formula (2)

The compound of the formula (3) of 6-allyl deriv preparation with acid treatment and dehydration, goes protection then in 2 ' is protected as embodiment 11, the compound of acquisition formula (2) as shown in Figure 1, wherein R _aBe OH, R _cBe H, R _fIt is allyl group.Similarly, as mentioned above, be used as the compound (R wherein of formula (1) _dAs above list) parent material prepared the compound (R wherein of formula (I) _dBe propyl group, butyl, benzyl, vinyl or 3-hydroxybutyl).

Embodiment 14

With 9=O changes into=NOH

According to the method for embodiment 6A, the carbonyl of 9 in erythromycin is changed into corresponding oxime.

Embodiment 15

-OR _fConversion

A. allyl group → propyl group: the logical nitrogen of ethanolic soln to any above compound (0.2 mmole) of preparation adds 10% palladium carbon (20 milligrams).To the logical hydrogen of mixture, spend the night then at hydrogen direct draught stirred reaction mixture.Filter reaction mixture, vacuum concentration obtains glassy mass.Silica gel column chromatography (95: 5: 0.5 methylene chloride-methanol-ammonia) obtains white solid state propylated compound.

B. allyl group → CH ₂CHO: to methylene dichloride (100 milliliters) solution of any above-mentioned compound that obtains (4.0 mmole)-78 ℃ of logical ozone 45 minutes.Then reaction mixture was led to nitrogen 10 minutes.Add dimethyl sulphides (1.46 milliliters, 20 mmoles) at-78 ℃, 0 ℃ of stirred reaction mixture 30 minutes.The reaction mixture vacuum concentration obtains white foam shape thing, and it need not purifying and just can use.The solution of THF (40 milliliters, 4.0 mmoles) by 55 ℃ of heating compounds and triphenyl phosphine (2.62 grams, 10.0 mmoles) 2.5 hours, the vacuum concentration reaction mixture obtains white foam.Silica gel column chromatography (1: 1 acetone-hexane, 75: 25: 0.5 acetone-hexane-triethylamines then) obtains required white solid state compound.

C. allyl group → CH ₂CH=NOH: the compound that in B, prepares (R wherein _aBe-CH ₂CHO, 0.08 mmole) add triethylamine (31 microlitres, 0.225 mmole) and hydroxylamine hydrochloride (7.7 milligrams, 0.112 mmole) in methyl alcohol (5 milliliters) solution, stirred reaction mixture is 6 hours at ambient temperature.Reaction mixture is placed ethyl acetate,, use dried over sodium sulfate, and vacuum concentration obtains fining glass shape thing with 5% sodium bicarbonate aqueous solution and salt water washing.Chromatography on silica gel (95: 5: 0.5 methylene chloride-methanol-ammonia) obtains the white solid state compound.

D.-CH ₂CH=NOH →-CH ₂CN: under nitrogen, in THF (5 milliliters) solution of the compound (0.267 mmole) that C prepares, add di-isopropyl carbodiimide (83 microlitres, 0.534 mmole) and CuCl (2.7 milligrams, 0.027 mmole), stirred reaction mixture spends the night at ambient temperature.Reaction mixture is placed ethyl acetate, with 5% sodium bicarbonate aqueous solution and salt water washing, use dried over sodium sulfate, vacuum concentration obtains fining glass shape thing.Silica gel column chromatography (95: 5: 0.5 methylene chloride-methanol-ammonia) obtains required white solid state compound.

E.-CH ₂CHO →-CH ₂CH ₂NH ₂: add ammonium acetate (212 milligrams, 2.76 mmoles) in methyl alcohol (10 milliliters) solution of the compound that in B, prepares (0.276 mmole), cooling mixture to 0 ℃.Add sodium cyanoborohydride (34 milligrams, 0.553 mmole), 0 ℃ of stirred reaction mixture 30 hours.Reaction mixture is placed ethyl acetate,, filter with 5% aqueous sodium carbonate, 2% 3 (hydroxymethyl) the aminomethane aqueous solution and salt water washing, dried over sodium sulfate, and vacuum concentration.Silica gel column chromatography (90: 10: 0.5 methylene chloride-methanol-ammonia) obtains required white solid state compound.

F.-CH ₂CHO →-CH ₂CH ₂NHCH ₂-phenyl: add acetate (114 microlitres, 2.00 mmoles) and benzylamine (218 microlitres, 2.00 mmoles) in methyl alcohol (10 milliliters) solution of the compound that in 0 ℃ B, prepares (0.200 mmole), stirred the mixture 10 minutes.Add sodium cyanoborohydride (24.8 milligrams, 0.400 mmole), stirred reaction mixture 16 hours.Add extra sodium cyanoborohydride (24.8 milligrams, 0.400 mmole) then, continue to stir 5 hours.Reaction mixture is placed ethyl acetate,, use dried over sodium sulfate, filter and vacuum concentration with 5% aqueous sodium carbonate, 2% 3 (hydroxymethyl) the aminomethane aqueous solution and salt water washing.Silica gel column chromatography (95: 50: 0.5 methylene chloride-methanol-ammonia), chromatography (50: 50: 0.5 acetone-hexane-triethylamines) obtains required white foam for the second time then.

G.-CH ₂CHO →-CH ₂CH ₂NHCH ₂CH ₂-phenyl: add acetate (114 microlitres, 2.00 mmoles) and styroyl amine (218 microlitres, 2.00 mmoles) in methyl alcohol (10 milliliters) solution of the compound that in 0 ℃ B, prepares (0.200 mmole), stirred the mixture 10 minutes.Add sodium cyanoborohydride (24.8 milligrams, 0.400 mmole), stirred reaction mixture 16 hours.Reaction mixture is placed ethyl acetate,, use dried over sodium sulfate, filter and vacuum concentration with 5% aqueous sodium carbonate, 2% 3 (hydroxymethyl) the aminomethane aqueous solution and salt water washing.Silica gel column chromatography (90: 10: 0.5 methylene chloride-methanol-ammonia) obtains required compound.

H.-CH ₂CHO →-CH ₂CH ₂NHCH (CO ₂CH ₃) CH ₂-phenyl: add L-phenylalanine methyl ester hydrochloride (129 milligrams, 0.600 mmole) in methyl alcohol (10 milliliters) solution of the compound that in 0 ℃ B, prepares (0.200 mmole), stirred the mixture 10 minutes.Add sodium cyanoborohydride (924.8 milligrams, 0.400 mmole), stirred reaction mixture 22 hours.Reaction mixture is placed ethyl acetate,, use dried over sodium sulfate, filter and vacuum concentration with 5% aqueous sodium carbonate, 2% 3 (hydroxymethyl) the aminomethane aqueous solution and salt water washing.Silica gel column chromatography (95: 5: 0.5 methylene chloride-methanol-ammonia) obtains required compound.

I.-CH ₂CHO →-CH ₂CH ₂NHCH ₂-(4-pyridyl): the method according to G has prepared required compound, except replacing phenylethylamine with the 4-aminomethyl pyridine.

J.-CH ₂CHO →-CH ₂CH ₂NHCH ₂-(4-quinolyl): add (23 milligrams of 4-quinoline aldehydes in methyl alcohol (2 milliliters) solution of the compound that in E, prepares (0.15 mmole), 0.15 acetate (8.6 microlitres mmole),, 0.15 mmole) and sodium cyanoborohydride (9.4 milligrams, 0.15 mmole), stirred the mixture 15 hours.Reaction mixture is placed ethyl acetate,, use dried over sodium sulfate, filter and vacuum concentration with 5% aqueous sodium carbonate, 2% 3 (hydroxymethyl) the aminomethane aqueous solution and salt water washing.Silica gel column chromatography (95: 10: 0.5 methylene chloride-methanol-ammonia) obtains required compound.

K. allyl group →-CH ₂CH=CH-phenyl: (22 milligrams of compound (1.00 mmole), the acid chlorides (II) of 2 ' of preparation protection among the embodiment 10 under nitrogen; 0.100 mmole) and (52 milligrams of triphenyl phosphines; 0.200 in acetonitrile mmole) (5 milliliters) solution; add iodobenzene (220 microlitres; 2.00 mmole) and triethylamine (280 microlitres; 2.00 mmole), mixture is cooled to-78 ℃, the degassing and sealing.With reaction mixture temperature to 60 ℃ 0.5 hour, 80 ℃ were stirred 12 hours, and placed ethyl acetate then, and with 5% sodium bicarbonate aqueous solution washed twice, 2% 3 (hydroxymethyl) aminomethane solution washing once, with the salt water washing once, use dried over sodium sulfate, filter and vacuum concentration.Silica gel column chromatography (95: 5: 0.5 methylene chloride-methanol-ammonia) obtains required compound.

Protection is finished in heating in methyl alcohol.

Other embodiment of formula (1)-(3) (R wherein _bBe H, R _cBe H, R _aBe OH, Y and Z altogether=O, R _dBe propyl group, butyl, benzyl, vinyl, or the 3-hydroxybutyl) be those wherein R _fBe:

-CH ₂CH ₂CH ₂-phenyl;

-CH ₂CH=CH-(4-p-methoxy-phenyl);

-CH ₂CH=CH-(4-chloro-phenyl-);

-CH ₂CH=CH-(3-quinolyl);

-CH ₂CH ₂CH ₂OH；

-CH ₂C(O)OH；

-CH ₂CH ₂NHCH ₃；

-CH ₂CH ₂NHCH ₂OH；

-CH ₂CH ₂N(CH ₃) ₂；

-CH ₂CH ₂(1-morpholinyl);

-CH ₂C(O)NH ₂；

-CH ₂NHC(O)NH ₂；

-CH ₂NHC(O)CH ₃；

-CH ₂F；

-CH ₂CH ₂OCH ₃；

-CH ₂CH ₃；

-CH ₂CH＝CH(CH ₃) ₂；

-CH ₂CH ₂CH(CH ₃)CH ₃；

-CH ₂CH ₂OCH ₂CH ₂OCH ₃；

-CH ₂SCH ₃；

-cyclopropyl;

-CH ₂OCH ₃；

-CH ₂CH ₂F；

-CH ₂-cyclopropyl;

-CH ₂CH ₂CHO；

-C(O)CH ₂CH ₂CH ₃；

-CH ₂-(4-nitrophenyl);

-CH ₂-(4-chloro-phenyl-);

-CH ₂-(4-p-methoxy-phenyl);

-CH ₂-(4-cyano-phenyl);

-CH ₂CH＝CHC(O)OCH ₃；

-CH ₂CH＝CHC(O)OCH ₂CH ₃；

-CH ₂CH＝CHCH ₃；

-CH ₂CH＝CHCH ₂CH ₃；

-CH ₂CH＝CHCH ₂CH ₂CH ₃；

-CH ₂CH=CHSO ₂-phenyl;

-CH ₂C≡CSi(CH ₃) ₃

-CH ₂C≡CCH ₂CH ₂CH ₂CH ₂CH ₂CH ₃；

-CH ₂C≡CCH ₃；

-CH ₂-(2-pyridyl);

-CH ₂-(3-pyridyl);

-CH ₂-(4-pyridyl);

-CH ₂-(4-quinolyl);

-CH ₂NO ₂；

-CH ₂C(O)OCH ₃；

-CH ₂C (O)-phenyl;

-CH ₂C(O)CH ₂CH ₃；

-CH ₂Cl；

-CH ₂S (O) ₂-phenyl;

-CH ₂CH＝CHBr；

-CH ₂CH=CH-(4-quinolyl);

-CH ₂CH ₂CH ₂-(4-quinolyl);

-CH ₂CH=CH-(5-quinolyl);

-CH ₂CH ₂CH ₂-(5-quinolyl);

-CH ₂CH=CH-(4-benzoxazolyl); Or

-CH ₂CH=CH-(7-benzimidazolyl-).

Above-mentioned any compound can change into corresponding derivative with embodiment 14 described methods above, and wherein Y and Z are=NOH altogether.

Embodiment 16

Fluoridizing of C2 position

2 '-O-benzoyl-6-O-propargyl-3-goes cladinosyl group-3-oxo-10, and 11-is anhydrous-2-fluoro-15-erythromycin A

Under inert atmosphere, 2 '-O-benzoyl-6-O-propargyl-3-is gone cladinosyl group-3-oxo-10,11-is anhydrous-and the tetrahydrofuran solution of 15-methyl-Erythromycin A is cooled to-78 ℃, handles with the tetrahydrofuran solution of 1.0M potassium tert.-butoxide.Stirred the mixture 5 minutes, and divided the tetrahydrofuran solution of 3 parts with 2 hours adding N-fluorobenzene sulfimides.After adding, make the reactant temperature, placed 5 hours in addition to envrionment temperature.Add K ₂CO ₃The aqueous solution is used CH ₂Cl ₂The extracting mixture.Merge organic extract, use MgSO ₄Drying is filtered and evaporation.Chromatography obtains product on silica gel.

Embodiment 17

Deriving of C-13 position

Parent material: 15-amido Erythromycin A diacetin

Handle 50 ml methanol solution of 15-azido-Erythromycin A (7.75 restrain 10 mmoles) with acetate (2.0 milliliters) and 10% palladium carbon (0.1 gram), under 1 atmospheric hydrogen, stir, disclose parent material up to thin layer chromatography analysis and reduce fully.With the suspension filtration over celite, remove catalyzer, be evaporated to the dried product that obtains then, it is as following deutero-parent material.

A.15-(quinolyl-4 kharophen) Erythromycin A is synthetic

Use quinolyl-4 Acetyl Chloride 98Min. (350 milligrams) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

B.15-(3-(quinolyl-4) propionamido) Erythromycin A is synthetic

Use 3-(quinolyl-4) propionyl chloride (400 milligrams) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

C.15-(isoquinoline 99.9-4-base kharophen) Erythromycin A is synthetic

Use isoquinoline 99.9-4-base Acetyl Chloride 98Min. (350 milligrams) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

D.15-(3-(isoquinoline 99.9-4-yl) propionamido) Erythromycin A is synthetic

Use 3-(isoquinoline 99.9-4-yl) propionyl chloride (400 milligrams) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

E.15-((quinoline-5-base is amino) kharophen) Erythromycin A is synthetic

Use (quinoline-5-base is amino) acetate (0.30 gram), dicyclohexyl carbodiimide (0.4 gram), I-hydroxybenzotriazole (0.25 gram) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

F.15-((quinoline-6-base is amino) kharophen) Erythromycin A is synthetic

Use (quinoline-6-base is amino) acetate (0.30 gram), dicyclohexyl carbodiimide (0.4 gram), I-hydroxybenzotriazole (0.25 gram) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

G.15-((quinolyl-4 methyl) carbamyl amino) Erythromycin A is synthetic

Use quinoline-4-methoxycarbonyl chlorine (400 milligrams) and triethylamine (0.5 milliliter) at 0 ℃ of 10 milliliters of dichloromethane solution handling 15-amido Erythromycin A diacetin (1.0 gram) successively.After 3 hours,, use saturated NaHCO with methylene dichloride diluting reaction thing ₃Solution washing three times.Use MgSO ₄Dry organic phase is filtered and evaporation, obtains crude product.The silica gel column chromatography purifying obtains pure products.

Claims

1. a compound is characterized in that, this compound has formula:

Or

Or its 10, the 11-dehydrated form;

Wherein

R _aBe H or OH;

R _bBe H or halogen;

R _cBe H or blocking group;

R _eIt is H or blocking group or single-or dibasic aminocarboxyl;

Z and Y be altogether=O ,=NOH or deutero-oxime;

2. compound as claimed in claim 1 is characterized in that R _dBe methyl, propyl group or vinyl.

3. compound as claimed in claim 1 is characterized in that R _fBe aromatic yl alkenyl or aromatic yl polysulfide yl.

4. compound as claimed in claim 3 is characterized in that R _fBe 3-arylprop-2-thiazolinyl or 3-aryl Propargyl.

5. compound as claimed in claim 4, it is characterized in that described aryl is 3-quinolyl, 4-quinolyl or 5-quinolyl, phenyl, 4-fluorophenyl, 4-chloro-phenyl-, 4-p-methoxy-phenyl, 6-quinolyl, 6-quinoxalinyl, 6-amino-3-quinoxalinyl or 4-isoquinolyl.

6. compound as claimed in claim 1 is characterized in that R _fBe H or C1-C3 alkyl.

7. compound as claimed in claim 6 is characterized in that R _fIt is methyl.

8. compound as claimed in claim 1 is characterized in that R _bIt is fluorine.

9. a pharmaceutical composition is characterized in that, said composition contains acceptable vehicle on described compound of claim 1 and the pharmacology.

10. the method for a control infection in object is characterized in that, this method comprises the described compound of claim 1 or its pharmaceutical composition of the object significant quantity that is applied to this control of needs.

11. a protection material to avoid the septic method of microorganism, it is characterized in that this method comprises the described compound of claim 1 that described material is used significant quantity.