CN1360593A - Pseudomycin N-acyl side-chain analogs - Google Patents

Abstract

Semi-synthetic pseudomycin compounds having structure (I) are described which may be useful as antifungal agents or intermediates in the design of antifungal agents.

Description

Pseudomycin N-acyl side-chain analogs

Invention field

The present invention relates to pseudobactin (pseudomycin) compound, especially have the semisynthetic pseudobactin compound of new N-acyl side-chain.

Background of invention

Pseudobactin is an isolating natural product from the liquid culture of pseudomonas syringae (Pseudomonas syringae, the bacterium relevant with plant), and has shown to have anti-mycotic activity.(for example referring to, Harrison, L. etc., " Pseudomycin; gang derives from the new peptide with broad-spectrum antifungal activity of pseudomonas syringae " J.Gen.Microbiology, 137 (12), 2857-65 (1991) and U.S. Pat 5,576,298 and 5,837,685).Different with the antimycotic agent (for example syringomycin, pseudomonas syringae toxin and pseudomonas syringae statin) that derives from pseudomonas syringae noted earlier, pseudobactin A-C contains hydroxyl aspartic acid, aspartic acid, Serine, dehydrogenation aminobutyric acid, Methionin and DAB.The peptide moiety of pseudobactin A, A ', B, B ', C, C ' is corresponding to the L-Ser-D-Dab-L-Asp-L-Lys-L-Dab-L-aThr-Z-Dhb-L-Asp with terminal carboxyl(group) (3-OH)-L-Thr (4-Cl), and this carboxyl will encircle closure greatly on the OH of the terminal Ser of N-.These analogues are distinguished by the N-acyl side-chain; be that pseudobactin A is by 3; 4-dihydroxyl tetradecanoyl N-acidylate; pseudobactin A ' is by 3; 4-dihydroxyl pentadecanoyl N-acidylate; pseudobactin B is by 3-hydroxyl tetradecanoyl N-acidylate; pseudobactin B ' is by 3-hydroxyl lauroyl N-acidylate; pseudobactin C is by 3; 4-dihydroxyl hexadecanoyl N-acidylate, and pseudobactin C ' is by 3-hydroxyl hexadecanoyl N-acidylate.(for example referring to Ballio, A. waits the people, " derive from the bioactive lipid depsipeptides of pseudomonas syringae: Pseudomycin, " FEBS Letters, 355 (1), 96-100, (1994) and Coiro, V.M. waits the people, " use derives from the geometry distance of positions of NMR data and the comformation in solution that molecular dynamics is passed through the false mycin A of the quasi-definite pseudomonas syringae MSU of computer mould 16H phytotoxicity fat depsipeptides; " Eur.J.Biochem., 257 (2), 449-456 (1998)).

Known pseudobactin has certain biological side effect.For example, when pseudobactin during, observed the local toxicity of venous endothelial destruction, disorganization, inflammation and host tissue through intravenous administration.Therefore, need from this compounds, identify treatment fungi infestation useful and do not have a compound of present observed side effect.

The invention summary

The invention provides represented pseudobactin compound, its pharmaceutically useful salt and the solvate thereof of following structural formula that is used as anti-mycotic agent or is used for the anti-mycotic agent design,

Wherein R is

Wherein

R ^aAnd R ^{A '}Be hydrogen or methyl, perhaps R independently ^aOr R ^{A '}Be alkylamino, with R ^bOr R ^{B '}Form 6-unit cycloalkyl ring, 6-unit's aromatic ring or two key together, perhaps with R ^cForm 6-unit aromatic ring together;

R ^bAnd R ^{B '}Be hydrogen, halogen or methyl, perhaps R independently ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl, condition is: work as R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydrogen and R ^fBe n-hexyl, n-octyl or positive decyl, perhaps R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydroxyl and R ^fDuring for n-octyl, n-nonyl or positive decyl, R ^{B '}It or not hydroxyl;

R ^cBe hydrogen, hydroxyl, C ₁-C ₄Alkoxyl group, hydroxy alkoxy base or and R ^eForm 6-unit's aromatic ring or C together ₅-C ₆Cycloalkyl ring;

R ^eBe hydrogen, perhaps with R ^fForm 6-unit aromatic ring, C together ₅-C ₁₄6-unit's aromatic ring or C that alkoxyl group replaces ₅-C ₁₄6-unit's aromatic ring that alkyl replaces and

R ^fBe C ₈-C ₁₈Alkyl, or C ₅-C ₁₁Alkoxyl group; Or R is

Wherein

R ^gBe hydrogen or C ₁-C ₁₃Alkyl, and

R ^hBe C ₁-C ₁₅Alkyl, C ₄-C ₁₅Alkoxyl group, (C ₁-C ₁₀Alkyl) phenyl ,-(CH ₂) _n-aryl or-(CH ₂) _n-(C ₅-C ₆Cycloalkyl), n=1 or 2 wherein; Perhaps R is

Wherein

R ⁱBe hydrogen, halogen or C ₅-C ₈Alkoxyl group, and

M is 1,2 or 3; R is

Wherein

R ^jBe C ₅-C ₁₄Alkoxyl group or C ₅-C ₁₄Alkyl and p=0,1 or 2; R is Wherein

R ^kBe C ₅-C ₁₄Alkoxyl group, perhaps R is-(CH ₂)-NR ^m-(C ₁₃-C ₁₈Alkyl), R wherein ^mBe hydrogen ,-CH ₃Or-C (O) CH ₃

In another embodiment of the present invention, a kind of medicinal preparations is provided, it comprises the pseudobactin compound and the pharmaceutically useful carrier of said structure I representative.

In another embodiment of the present invention, a kind of method for the treatment of anti-fungal infection in the animal of needs treatment is provided, this method comprises to described animal uses above-mentioned pseudobactin Compound I.

In another embodiment of the present invention, a kind of method for preparing pseudobactin compound unhindered amina nuclear is provided, this amine is endorsed the compound that is generated said structure I representative by acidylate.This method comprises pseudobactin compound (for example, pseudobactin A, A ' or step C) that contains the N-acyl group alkyl group side chain that has a γ or δ hydroxyl at least with trifluoroacetic acid or acetic acid treatment.

Definition

Term used herein " unhindered amina pseudobactin nuclear " or " pseudobactin nuclear " are meant the structure of following I-A:

Wherein, R ' is-NH ₂Or-NH _p-P _g, P wherein _gFor amino protecting group and p are 0 or 1.

Term " alkyl " refers to the general formula C that contains 1-30 carbon atom _nH _2n+1Alkyl.Alkyl can be straight chain (for example methyl, ethyl, propyl group, butyl etc.), side chain (for example sec.-propyl, isobutyl-, the tertiary butyl, neo-pentyl etc.), ring-type (for example cyclopropyl, cyclobutyl, cyclopentyl, methylcyclopentyl, cyclohexyl etc.) or polycyclic (for example two ring [2.2.1] heptane, spiral shell [2.2] pentane etc.).Described alkyl can be substituted or not replace.Similarly, the moieties in alkoxyl group, alkyloyl or the alkanoates has top identical definition.

Term " thiazolinyl " refers to the acyclic hydrocarbous that contains at least one carbon-carbon double bond.Described thiazolinyl can be straight chain, side chain, ring-type or polycyclic.Described thiazolinyl can replace or not replace.Alkenyl part in alkene oxygen base, enoyl-or the olefin(e) acid ester has top identical definition.

Term " alkynyl " is meant and contains a carbon carbon triple-linked acyclic hydrocarbous at least.Alkynyl can be straight chain or side chain, can be that replace or unsubstituted.Alkynyl in alkynyloxy group, alkynes acyl group or the acetylenic acid ester group partly has implication same as described above.

Term " aryl " refers to the aromatics part of have the monocycle system (for example phenyl) or condensed ring system (for example naphthalene, anthracene, phenanthrene etc.).Described aromatics can be substituted or not replace.

Term " heteroaryl " refers to contain at least a heteroatomic aryl (for example pyrroles, pyridine, indoles, thiophene, furans, cumarone, imidazoles, pyrimidine, purine, benzoglyoxaline, quinoline etc.) in aromatic ring system.Aryl can comprise list or condense ring system.Heteroaryl can be that replace or unsubstituted.

" NH _p-P _g" and " amino protecting group " be meant and be commonly used for amino substituting group to stop when other functional group reactions of compound or the protection amido functional group.When p was 0, the coupled nitrogen of amino protecting group formed cyclin imide together, for example phthalic imidine and tetrachloro-phthalimide.When p was 1, the nitrogen that amino protecting group can be coupled formed carbamate together, for example methyl, ethyl and 9-fluorenyl methyl carbamate; Or acid amides, for example N-formyl radical and N-ethanoyl acid amides.

In organic chemistry filed, particularly organic biochemical field should be interpreted as widely, and effective replacement of compound allows, perhaps or even useful.For example, in the present invention, the term alkyl allows to comprise typical alkyl substituent, for example methyl, ethyl, propyl group, hexyl, iso-octyl, dodecyl, stearyl (stearyl) etc.Term " group " is specific to be related to and allows to be included on the alkyl of this area routine and replace, for example hydroxyl, halogen, alkoxyl group, carbonyl, ketone group, ester group, carbamate groups (Carbamato) etc., and comprise there is not the moieties that replaces.Yet those skilled in the art are generally understood as, and substituting group should be through selection, so that the pharmacological property of compound is not had negative impact, perhaps the use to this medicine does not have negative interference.The suitable substituent of any group defined above comprises alkyl, thiazolinyl, alkynyl, aryl, halogen, hydroxyl, alkoxyl group, aryloxy, sulfydryl, alkylthio, arylthio ,-and two-alkylamino, quaternary ammonium salt, aminoalkoxy, hydroxyalkyl amino, amino alkylthio, formamyl, carbonyl, carboxyl, glycolyl, glycyl, diazanyl, amidino groups and combination thereof.

Term " solvate " refers to aggregate, and it comprises one or more solute molecules, for example the compound of structure I and one or more acceptable solvent molecule, for example water and ethanol etc.

Term " but medication salt " is meant the organic or inorganic salt of the compound of structure I representative, and it is nontoxic substantially at dosage to the recipient.

Term " animal " refers to people, pet (for example dog, cat and horse), food source animal (for example ox, pig, sheep and poultry), zoo animal, marine animal, birds and other similar animal kind.

Detailed Description Of The Invention

The applicant finds the unitary N-acyl group of the L-Serine deacylation of pseudobactin compound; obtain new compound with new N-acyl group acidylate again subsequently; experiment in vitro show these new compounds have anti-candida albicans (C.albican), novel Cryptococcus (C, neoformans) and/or Aspergillus fumigatus (Aspergillus fumigatus) activity.

Following reaction scheme I represents the general method by any naturally occurring pseudobactin synthetic compound I.A kind of naturally occurring pseudobactin compound though drawn in reaction scheme I it should be appreciated by those skilled in the art that the side chain of the semi-synthetic derivative of naturally occurring pseudobactin compound is modified and can be finished by similar approach.Usually, it is synthetic that the preparation Compound I needed for four steps: (1) selectivity amido protecting; (2) the chemical deacylation of N-acyl side-chain or enzymatic deacylation; (3) with different side chain acidylate again; (4) described amino deprotection. Reaction scheme I

Standard method at the amino available any protection amino well known by persons skilled in the art of 2,4 and 5 side chains is protected.The concrete kind of amino protecting group is not really important; as long as deutero-amino stable and protecting group under the reaction conditions that takes place subsequently on other position of intermediate molecule can selectivity be removed and do not influenced the rest part of molecule in the suitable time, comprise that any other amino protecting group gets final product.Suitable amino protecting group comprises benzyl oxygen base carbonyl, to nitrobenzyl oxygen base carbonyl, to bromobenzyl oxygen base carbonyl, to methoxy-benzyl oxygen base carbonyl, p-methoxyphenyl azo benzyl oxygen base carbonyl, to phenylazo benzyl oxygen base carbonyl, tertiary butyl oxygen base carbonyl, cyclopentyloxy carbonyl and phthaloyl imino.Preferred amino protecting group is tert-butoxycarbonyl (t-Boc), allyl group oxygen base carbonyl (Alloc), phthaloyl imino and benzyl oxygen base carbonyl (CbZ or CBZ).Most preferably be allyl group oxygen base carbonyl and benzyl oxygen base carbonyl.In addition, suitable protecting group is described in T.W.Greene " Protective Groupsin Organic Synthesis " John Wiley and Sons, New York, N.Y., (2nd ed., 1991), the 7th chapter.

Deacylation with N-acyl group of the hydroxylated side chain of γ or δ (for example, 3,4-ipurolic acid ester (tetradeconoate)) can be finished by the pseudobactin compound of handling amido protecting with the 5-20% aqueous acid.Suitable acid comprises acetate and trifluoroacetic acid.Preferred acid is trifluoroacetic acid.If use trifluoroacetic acid, this reaction can be finished in room temperature or near under the room temperature.But when using acetate, this reaction is generally carried out at about 40 ℃.The dissolving of available a kind of organic solvent promotion pseudobactin compound that can be water-soluble.Suitable water solvent system comprises acetonitrile, water and its mixture.Acetonitrile is particularly useful when the pseudobactin compound of protection goes acidylate.The preferred acid solution of pseudobactin compound deacylation that is used to protect is the acetonitrile solution of 8% trifluoroacetic acid.Organic solvent quickens this reaction, yet, add organic solvent and can cause other by product.The pseudobactin compound (for example, pseudobactin B and C ') that lacks the δ hydroxyl on the side chain can be through the enzymatic deacylation.Suitable deacylation enzyme comprises the polymyxin acylase, and (164-16081 fat acylase (crude product) or 161-16091 fat acylase (pure product) can be available from Wako Pure Chemical Industries; Ltd.); or ECB deacylation enzyme (referring to; U.S.Patent No.5 for example; 573,936).The enzymatic deacylation can be finished with standard deacylation method well known to those skilled in the art.For example, see Yasuda, N. etc., Agric.Biol.Chem., 53,3245 (1989) and Kimura, Y. etc., Agric.Biol.Chem., 53,497 (1989) with the general method of polymyxin acylase.

Deacylation product (be also referred to as pseudobactin nuclear or " PSN ") is with the respective acids of purpose acyl group acidylate again in the presence of the carbonyl activator." carbonyl activating group " is meant the substituting group of the nucleophilic addition of promotion on this carbonyl.Suitable active substituent is to have those that draw electronic action only on the carbonyl.These groups comprise, but be not limited to, alkoxyl group, aryloxy, nitrogenous aromatic heterocycle or amino (for example, oxygen base benzotriazole, imidazolyl, nitro-phenoxy, pentachloro-phenoxy group, N-oxygen base succinimide, N, N '-dicyclohexyl isourea-O-base and N-hydroxy-n-methoxyl group amino); Acetic ester; Manthanoate; Sulphonate (for example, methane sulfonate, ethane sulfonic acid ester, benzene sulfonate and p-toluenesulfonic esters) and halogenide (for example, muriate, bromide and iodide).

Perhaps, available solid phase synthesis wherein uses hydroxybenzotriazole resin (HOBt-resin) to be used for this acylation reaction as coupling agent.

In acylation process, can use various acid.Suitable acid comprises and contains one or more side chain aryl, alkyl, amino (comprise primary, the second month in a season and tertiary amine), hydroxyl, alkoxyl group and amino aliphatic acid; The aliphatic acid that in aliphatic chain, contains nitrogen or oxygen; By the aromatic acid of alkyl, hydroxyl, alkoxyl group and/or alkylamino replacement; With the heteroaromatic acid that is replaced by alkyl, hydroxyl, alkoxyl group and/or alkylamino.Acylate can be used as active anti-mycotic agent or is used as the intermediate of preparation active compound.Even some compounds are useful not as other compound, its activity profile also provides valuable information to the mentality of designing of desire acquisition optimum activity.

In case amino by acidylate, can be by in the presence of hydrogenation catalyst (for example 10%Pd/C), removing amino protecting group (2,4 and 5).When amino protecting group was allyl group oxygen base carbonyl, this protecting group can be removed with tri-n-butyltin hydride and triphenyl phosphine palladium dichloride.The advantage of this specific protection/deprotection route is the possibility that has reduced the unitary vinyl of hydrogenation pseudobactin Z-Dhb.

Discuss as previous, pseudobactin is from the isolating natural product of pseudomonas syringae, it is characterized as being ester depsipeptides (Lipodepsinonapetpides), it contains by the cyclic peptide part of lactone bond closed loop and comprises uncommon amino acid 4-chlorine Threonine (ClThr), 3-hydroxyl aspartic acid (HOAsp), 2,3-dehydrogenation-2-aminobutyric acid (Dhb) and 2,4-diamino-butanoic (Dab).Thereby the different strains of growth pseudomonas syringae is produced different pseudobactin analogue (A, A ', B, B ', C and C ') method be described below and be described in greater detail in the PCT patent application that is entitled as " producing pseudobactin (Pseudomycin Production by PseudomonasSyringae) " that people such as Hilton submitted on April 14th, 2000 by pseudomonas syringae, its sequence number is PCT/US00/08728, the PCT patent application that is entitled as " pseudobactin natural product (Pseudomycin Natural Products) " that people such as Kulanthaivel submitted on April 14th, 2000, its sequence number is PCT/US00/08727, and US 5,576,298 and 5,837, in 685, they all are incorporated herein by reference at this.

The pseudomonas syringae isolated strains that produces one or more pseudobactins is known in this area.Wild type strain MSU 174 and this bacterial strain mutant MSU 16H (ATCC 67028) that produces by transposon mutagenesis are described in US 5,576,298 and 5,837,685; People such as Harrison " Pseudomycins; a family of novel peptides fromPseudomonas syringae possessing broad-spectrum antifungalactivity; " J.Gen.Microbiology, 137,2857-2865 (1991); And people such as Lamb " Transposon mutagenesis and tagging offluorescent pseudomonas:Antimycotic production is necessaryfor control of Dutch elm disease; " Proc.Natl.Acad.Sci.USA, 84, among the 6447-6451 (1987).

Be fit to produce one or more pseudobactins the pseudomonas syringae bacterial strain can from the ambient source that comprises plant (for example barley plants, oranges and tangerines plant and cloves plant) and, for example separate in the source of soil, water, air and dust.Preferred strain is isolating from plant.Can be referred to as wild-type from the isolating pseudomonas syringae bacterial strain of ambient source.As used herein, " wild-type " is meant the natural dominant gene type (for example finding and be not the pseudomonas syringae bacterial strain or the isolate of chamber operation production by experiment at occurring in nature) that is present in the pseudomonas syringae normal microflora.Identical with most of organisms, the characteristic of the culture of used maternity leave unit cell rhzomorph (pseudomonas syringae bacterial strain such as MSU 174, MSU 16H, MSU 206,25-B1,7H9-1) is easy to change.Therefore, the offspring of these bacterial strains (for example recombinant chou, mutant and mutation) can obtain by methods known in the art.

Pseudomonas syringae MSU 16H is from the American Type CultureCollection, Parklawn Drive, and Rockville, MD, USA are public Ke De, its preserving number ATCC 67028 obtains.Pseudomonas syringae bacterial strain 25-B1,7H9-1 and 67 H1 were deposited in the American Type Culture Collection and have following preserving number respectively on March 23rd, 2000:

25-B1 preserving number PTA-1622

7H9-1 preserving number PTA-1623

67H1 preserving number PTA-1621

The mutants which had of pseudomonas syringae also is fit to produce one or more pseudobactins.As used herein, " mutant " is meant unexpected heritable variation in the bacterial strain phenotype, it can be spontaneous or pass through known mutagenic compound inductive, for example radiation (for example ultraviolet radiation or x-ray), chemical mutagen (for example ethylmethane sulfonate (EMS), diepoxy octane, N-methyl-N-nitro-N '-nitro guanine (NTG) and nitrous acid), location specific mutagenesis and transposon-mediated mutagenesis.The pseudomonas syringae mutant of maternity leave unit cell rhzomorph can be produced by handling this bacterium with the mutagenic compound that produce the amount of mutant effectively, shown in mutant excessively produce one or more pseudobactins, produce and a kind ofly surpass the pseudobactin (for example pseudobactin B) of other pseudobactin or under favourable growth conditions, produce one or more pseudobactins.Although the type of used mutagenic compound and quantity can change, preferred method is the level that can serially NTG be diluted to 1-100 μ g/ml.Preferred mutant be excessively produce pseudobactin B and in the substratum that minimum limits, grow those.

For following desired characteristic: habit, growth medium nutrition source, carbon source, growth conditions, amino acid need etc., can be to environment separation thing, mutants which had and other required Pseudomonas syringae bacterial strain of pseudomonas syringae through selecting.Preferably be chosen in for example growth and/or produce the pseudomonas syringae bacterial strain of one or more amounts on the N21 substratum of the minimum substratum that limits greater than the maternity leave unit cell rhzomorph of the pseudobactin of about 10 μ g/ml.Preferred strain is being when containing three kinds or amino acid still less, when growing on the optional substratum that contains lipid, potato product or its combination, presents the characteristic that produces one or more pseudobactins.

Use method as known in the art,, can cultivate recombinant bacterial strain by transforming the pseudomonas syringae bacterial strain.Except the microbiotic that these bacterial strains produce,, the conversion of pseudomonas syringae bacterial strain can be expressed different gene products by using recombinant DNA technology.For example, people can modify these bacterial strains, thereby introduce the endogenous pseudobactin biosynthesis gene of multiple copy, to obtain bigger pseudobactin output.

In order to produce one or more pseudobactins from pseudomonas syringae wild-type or mutants which had, this organism is containing three kinds of significant quantity or amino acid still less, stir culture in the water-based nutritional medium of preferred L-glutamic acid, glycine, Histidine or its combination.Perhaps, with glycine and one or more potato product and lipid combination.Can effectively grow and produce at pseudomonas syringae under the condition of required pseudobactin and cultivate.Condition for validity comprises that temperature is about 22 ℃-Yue 27 ℃, and the time is about 36 hours-Yue 96 hours.Oxygen concn in the culturing process of pseudomonas syringae in the control substratum is useful to the production of pseudobactin.The preferred oxygen level is maintained at about the 5-50% saturation ratio, more preferably from about 30% saturation ratio.Spray the oxygen concn to regulate in the substratum with air, pure oxygen or the gaseous mixture that contains aerobic.

The pH of control substratum also is useful in the culturing process of pseudomonas syringae.Pseudobactin is unstable under alkaline pH, and if the pH of substratum in about lasting about time more than 12 hours 6 or more, can produce tangible degraded.The pH of preferred culture medium remains between the 6-4.Pseudomonas syringae can produce one or more pseudobactins when growth in batch culture.Yet, add (fed-bath) or semicontinuous adding glucose and optional in batches, add acid or alkali (for example ammonium hydroxide) control pH and can improve output.Pseudobactin output can also use the continuous culture method of automatic adding glucose and ammonium hydroxide to improve.

The selection of pseudomonas syringae bacterial strain can influence the amount and the distribution of the pseudobactin of generation.For example, bacterial strain MSU 16H and 67 H1 mainly produce pseudobactin A separately, but also produce pseudobactin B and C, and its ratio typically is 4: 2: 1.The about 3-5 of height that the amount of the pseudobactin that bacterial strain 67 H1 typically produce is produced than bacterial strain MSU 16H doubly.Compare with 67 H1 with bacterial strain MSU 16H, bacterial strain 25-B1 produces more pseudobactin B and pseudobactin C still less.The characteristics of bacterial strain 7H9-1 are mainly to produce pseudobactin B, and the height of other bacterial strain of rate ratio of pseudobactin B.For example, this bacterial strain pseudobactin B that can produce is at least 10 of pseudobactin A or C.

Can every kind of pseudobactin, pseudobactin intermediate and mixture be detected, determine, separate and/or purify by any different methods well known by persons skilled in the art.For example, the pseudobactin in the composition of broth culture or isolate or purification or the activity level of pseudobactin can be determined by the anti-for example anti-mycotic activity of the fungi of mycocandida, and can separate and purification by high performance liquid chromatography.

This pseudobactin compound separation can be gone out and use with itself or with its pharmaceutically useful salt or solvate forms.Term " pharmaceutically useful salt " is meant the non-toxicity acid salt that is obtained by mineral acid and organic acid.The salt derivative that is fit to comprises halogenide, thiocyanate-, vitriol, hydrosulfate, sulphite, hydrosulphite, arylsulphonate, alkyl-sulphate, phosphoric acid salt, monohydric phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate salt, alkanoate, the cycloalkyl alkanoate, aryl-alkanoic salt, adipate, alginate, aspartate, benzoate, fumarate, the glucose enanthate, glycerophosphate, lactic acid salt, maleate, nicotinate, oxalate, palmitate, pectinic acid salt, picrate, Pivalate, succinate, tartrate, Citrate trianion, camphorate, camsilate, digluconate, trifluoroacetate etc.

Term " solvate " is meant the aggregate that contains one or more solute molecules (being the Pseudomycin prodrugs compound) and one or more medicinal solvent molecule such as water, ethanol etc.When solvent was water, this aggregate was referred to as hydrate.Solvate normally by with the prodrug heating for dissolving in suitable solvent and slowly cooling produce that amorphous or recrystallisation solvent thing form form.

Typically activeconstituents (being the pseudobactin derivative) being mixed with provides the drug dose that is easy to control and to the pharmaceutical dosage form of the product of a kind of elegance of patient, doctor or animal doctor and easy handling.Preparation can contain the activeconstituents of 0.1%-99.9%wt, and that more conventional is the about 30%wt of about 10%-.

As used herein, term " unitary dose " or " dose unit " are meant the physics discrete unit that contains the predetermined amount activeconstituents that produces required result of treatment as calculated.When unitary dose oral administration or parenterai administration, typically the form with determination unit in tablet, capsule, pill, powder packets, topical composition, suppository, wafer, ampoule or the multi-dose container etc. provides.Perhaps, the unitary dose drying that can suck or spray or the form administration of liquid aersol.

Dosage can change according to the severity of the physical characteristics of animal, animal illness and the mode that is used for administration.The concrete dosage of given animal is often determined by doctor or animal doctor's judgement.

Carrier, thinner and the vehicle that is fit to those skilled in the art for known and comprise following material: carbohydrate, wax, water-soluble and/or water-swelling polymer, hydrophilic or hydrophobic material, gelatin, oil, solvent, water etc.Used specific support, thinner or vehicle will depend on the use-pattern and the purpose of activeconstituents.Goods can also comprise wetting agent, lubricant, tensio-active agent, buffer reagent, toughener, weighting agent, stablizer, emulsifying agent, suspension agent, sanitas, sweeting agent, flavouring agent, seasonings and combination thereof.

Medicinal compositions can make the administration that ins all sorts of ways.The method that is fit to comprises part (for example ointment or sprays), oral, injection and sucks.Used concrete treatment process will depend on the type of the infection of treatment.

When non-enteron aisle intravenous applications, before administration, these goods are typically through dilution or preparation again (if through freeze dried words), further dilution if necessary.The example of the preparation explanation again of freeze-drying prods is to join 10ml water for injection (WFI) in the bottle and stirring and dissolving gently.Again the preparation time is lower than 1 minute to the typical case.Before administration, gained solution further is diluted in the transfusion as the water (D5W) of 5% glucose then.

The pseudobactin compound has shown to have anti-mycotic activity, for example comprises the infectivity fungi growth that suppresses following: mycocandida various (being Candida albicans (C.albicans), Candida parapsilosis (C.parapsilosis), Crewe Si Shi candiyeast (C.krusei), Candida glabrata (C.glabrata), candida tropicalis (C.tropicalis) or Candida lusitaniae (C.lusitania)); Torulopsis is various (to be torulopsis glabrata (T.glabrata); Aspergillus various (being Aspergillus fumigatus (A.fumigatus)); Histoplasma various (being Histoplasma capsulatum (H.capsulatum)); Cryptococcus various (being novel Cryptococcus (C.neoformans)); Blastomyces various (being Blastomyces dermatitidis (B.dermatitidis)); Fusarium is various; Trichophyton is various, Pseudallescheria boydii, posadasis spheriforme, sporotrichum schenckii etc.

So compound of the present invention and preparation can be used for resisting in the medicine of systemic fungal infection or fungal skin infections useful in preparation.Therefore, provide a kind of method that suppresses fungi activity, this method comprises Compound I of the present invention is contacted with fungi.Preferred method comprises inhibition Candida albicans, novel Cryptococcus or activity of Aspergillus fumigatus.Term " contact " comprises combining or joint of The compounds of this invention and fungi, perhaps surface contact or be in contact with one another.This term is not given any other restriction to this method, for example by suppressing mechanism.These methods are defined as and comprise by the effect of these compounds and the activity of inherent anti-fungal property inhibition fungi thereof.

A kind of method for the treatment of fungi infestation also is provided, has comprised pharmaceutical preparation of the present invention the host animal effective dosage of this treatment of needs.Preferred method comprises treatment Candida albicans, novel Cryptococcus or infection by Aspergillus fumigatus.Term " significant quantity " is meant the amount of the active compound that can suppress fungi activity.Dosage changes with for example following factor: the character of infection and severity, host's age and general health, host are to the tolerance of anti-mycotic agent and host's kind.Concrete dosage can change according to these factors equally.Medicine can single per daily dose or the mode of the multidose during a day use.This scheme can extend to about 2-3 week or longer from about 2-3 days.Typical per daily dose (with single or divide equally dosed administration) contains the dosage level of the active compound of the 0.01mg/kg-100mg/kg body weight of having an appointment.Preferred per daily dose is generally about 0.1mg/kg-60mg/kg, more preferably about 2.5mg/kg-40mg/kg.The host can be any animal, comprises people, pet (for example dog, cat and horse), food source animal (for example ox, pig, sheep and poultry), zoo animal, marine animal, birds and other similar animal species.

Embodiment

Except as otherwise noted, all chemical reagent available from Aldrich Chemical (Milwaukee, WI).

Biological sample

Pseudomonas syringae MSU 16H is from American type culture collection (AmericanType Culture Collection), Parklawn Drive, and Rockville, MD, USA public Ke De's, preserving number is ATCC 67028.Pseudomonas syringae bacterial strain 25-B1,7H9-1 and 67 H1 are preserved in American type culture collection on March 23rd, 2000, and preserving number is as follows:

The 25-B1 preserving number is PTA-1622

The 7H9-1 preserving number is PTA-1623

The 67H1 preserving number is the PTA-1621 chemical abbreviations

In used embodiment, represent the material listed respectively with following abbreviation:

The ACN-acetonitrile

The TFA-trifluoroacetic acid

The DMF-dimethyl formamide

DEAD-azoethane dicarboxylic acid esters

EDCI-1-[3-(dimethylamino) propyl group]-the 3-ethyl-carbodiimide hydrochloride

The BOC=tert-butoxycarbonyl, (CH ₃) ₃C-O-C (O)-

CBZ=benzyl oxygen base carbonyl, C ₆H ₅CH ₂-O-C (O)

FMOC=fluorenyl methyl oxygen base carbonyl

The HPLC condition

Except as otherwise noted, analyzing the reversed-phase HPLC operation is that (C18, the Waters 600E system of 3.9 * 300mm) posts carries out with assembling Waters μ Bondapak.Used eluent is 65: 35 acetonitrile/0.1%TFA water solvent system-100% acetonitriles (with 20 minutes), and flow velocity is 1.5ml/ minute, and uses UV to detect at 230nm.

It is to carry out with the Waters Prep2000 system that adopts Dynamax 60 dust C18 posts that preparation HPLC handles, and wherein use and the identical solvent system of analysis HPLC system, but flow velocity is 40ml/ minute.

The detection of bioanalysis anti-mycotic activity and quantitative assay:

Come at the external test anti-mycotic activity by the minimum inhibition concentration (MIC) that uses test of standard agar dilution or disk to launch test acquisition compound.The typical fungus of using in the anti-mycotic activity test is a Candida albicans.(50 μ l) thinks when the inhibition that is inoculated into the Candida albicans on the agar plate is caused the 10-12mm diameter region that remarkable anti-mycotic activity is arranged when test sample book.The tail venous toxicity:

At the 0th, 24,48 and 72 hour, treat mouse via lateral tail vein intravenously (IV) administration with 0.1ml test compounds (20mg/kg).Every group comprises 2 mouse.Compound is formulated in 0.5% glucose and the sterile water for injection.Treatment back monitoring mouse is 7 days first, and examine comprise erythema, swelling, variable color, necrosis, tail in being lost in the stimulation symptom and show toxic other side effect symptom.

The mouse of using in this experiment is outbreed, the mean body weight of male ICR mouse be 18-20g (derive from Harlan Sprangue Dawley, Indianapolis, IN).

General method is used for the general method at 2,4 and 5 protection side chain amino of pseudobactin A, A ', B, B ', C or C '

With pseudobactin compound (R ¹=H) dissolve/be suspended in (20mg/ml, Aldrich Sure Seal) among the DMF.Under room temperature, stirring, add N-(benzyloxycarbonyloxy) succinimide (6eq), at room temperature stirred 32 hours, by HPLC (4.6 * 50mm, 3.5 μ m, 300-SB, C8, Zorbax column) monitoring reaction.Rotation is evaporated to 10ml under room temperature, high vacuum, and this material is freezing until being ready to the chromatogram preparation.After anti-phase preparation HPLC and lyophilize, obtain unformed, white solid.Be used for L-Serine unit N-acyl group chemistry and remove the general method of acidylate.

The pseudobactin A of protection is dissolved/is suspended in water/acetonitrile (2: 1 H ₂O: ACN, about 3.5mg/ml) in and at room temperature slowly add TFA (8% volume).At room temperature stirring reaction is intact up to raw material consumption.Under room temperature, vacuum, remove acetonitrile and lyophilize material.The solid that obtains is dissolved among a small amount of DMF (if needs add water, adding the ACN of equivalent then), after preparation HPLC and lyophilize, generally obtains white, unformed solid (may be tfa salt).The pseudobactin nuclear solid phase acidylate of HOBt resin.The following example tetradecanoic acid; Yet for other organic acid, also available same general method.

In the reaction tubes of a 100ml two ends glass sintering, (1.03g 3.62mmol) is dissolved among 1: 1 DMF/THF of 50ml with tetradecanoic acid.In this solution, add resin HOBt (1.94g, 2.9mmol) and EDCI (0.556g, 2.9mmol) and jolt and spend the night.The solvent of draining, resin is with 2xDMF, 2xTHF and 2x 1: 1DMF/THF washing.(1.0 grams 0.723mmol) are dissolved among the 50ml DMF/THF (20mg/ml), and are added in the Acibenzolar of this resin-bonded and at turner or jolt to mix on the device and spend the night with the pseudobactin nuclear of CBZ-protection.Product and resin separated and with 2xDMF, 2xTHF and 2x DMF/THF washing in 1: 1 residual resin.The filtrate that merges is separated the pseudobactin product that also lyophilize obtains the CBZ-protection of (129mg, 10%) tetradecanoyl acidylate through reversed-phase HPLC.The pseudobactin nuclear acidylate of Acibenzolar (HOBt-methanesulfonates).The following example glycine tetradecanoic acid, however also can use this general method for other organic acid.

In a 500ml round-bottomed flask, (0.309g 1.1mmol) is dissolved among the 100ml DMF with the glycine tetradecanoic acid.In this solution, add the HOBt-methanesulfonates (0.229g, 1.1mmol) and triethylamine (0.081g, 0.8mmol).At 1atm N ₂Stir this solution down fast and spend the night, drying is removed DMF and TEA under high vacuum.Remaining oily matter distills 3 times up to forming white solid with methylbenzene azeotropic.The pseudobactin nuclear that in this solid, adds 100ml DMF and 1g CBZ-protection.Stirred solution spends the night and is dry under high vacuum.Product obtains the pseudobactin product of the CBZ-protection of (233mg, 20%) myristoyl acidylate through reversed-phase HPLC purifying and lyophilize.Be used for through hydrogenation at the amino de-protected general method of 2,4 and 5 side chains

Be dissolved in the activated derivatives of CBZ-protection in 13% cold acetate/methanol solution (5mg/ml) and add normal 10%Pd/C.By outgasing and using H ₂Displaced volume fills hydrogen 4-7 time in reaction.At room temperature react, once intact with HPLC and per 15 minutes monitoring reactions of mass spectrograph up to raw material consumption.After reaction is finished, remove air bag also with filter disc (Acrodisk GHP, the GF by Gelman) filtering reacting liquid of 0.45 μ m, be concentrated to about 1/10 volume and use preparation HPLC purifying, lyophilize contains the fraction of product.The preparation of preparation side chain precursor (1c)

Between in the 250ml round-bottomed flask that contains 100ml degassing ACN, adding-bromobenzaldehyde (5.000g, 27.02mmol), triethylamine (5.490g, 54.25mmol) and the 1-dodecyne (5.000g, 30.06mmol).In this mixture, add PdCl ₂(243.1mg, 1.370mmol), triphenyl phosphine (718.8mg, 2.740mmol) and CuI (173.8mg, 0.9120mmol).Reflux and reaction are spent the night then.Then reaction solution is chilled to room temperature and removes under vacuum and desolvate, the resistates that obtains is handled with methylene dichloride and with 2 * 1N HCl and the water washing of 1X salt.Organic layer MgSO ₄Dry.Leach siccative, under vacuum, remove and desolvate, obtain the brown oily title compound of 3.73 grams through silica gel chromatography and with 3%EtOAc/ hexane wash-out.Spectroscopic data with-structure of (l-dodecyne base) phenyl aldehyde (la) is consistent.

(1.00g is 3.70mmol) with 0.1g 5%Pd/Al to add above-claimed cpd in 100 mL EtOAc ₂O ₃H at room temperature, 50psi ₂Following reaction mixture 1 hour.Removing catalyzer, diatomite is with a large amount of EtOAc rinsings through diatomite filtration for reaction mixture.Remove the product that EtOAc obtains 882.1mg through Rotary Evaporators.This product is used for next step without being further purified.Spectroscopic data with-(lb) is consistent for the dodecyl phenyl aldehyde.

Under nitrogen atmosphere ,-78 ℃, in the 50ml round-bottomed flask that oven dried is crossed, add the anhydrous THF of 6.0ml and add subsequently the di-isopropyl lithamide (heptane of 1.2mL 2M/THF/ ethylbenzene solution, 2.41mmol).(0.331ml 2.45mmol) and with the solution that obtains is warmed up to approximately-40 ℃ and kept 1 hour under this temperature to wherein adding tert.-butyl acetate.(501.9mg 1.83mmol) is dissolved in the anhydrous THF of 4mL and also is chilled to-40 ℃ solution in advance to drip above-claimed cpd then in this negatively charged ion.Stirred reaction mixture 1 hour is used saturated aqueous ammonium chloride solution of 2ml and 10ml water termination reaction then.Reaction mixture is distributed between ether and water, and organic layer with the salt water washing once and use dried over sodium sulfate.Leach siccative, under vacuum, remove and desolvate,, obtain the 238.1mg yellow oil with 5%EtOAc/ hexane wash-out through silica gel chromatography.Spectroscopic data is consistent with 3-hydroxyl-3-(dodecylbenzyl) propionic acid tert-butyl ester.

(0 ℃) 4ml TFA solution of precooling is added in the 50ml round-bottomed flask that contains crude product 3-hydroxyl-3-(dodecylbenzyl) propionic acid tert-butyl ester.Stirring reaction 25min under this temperature, TLC (10%EtOAC/ hexane) shows the starting ester completely consumed.Under vacuum, remove TFA and obtain oily matter (1c).The preparation of side chain precursor (2c):

Compound 2a identical method with above-mentioned synthetic compound 1a replaces the 1-dodecyne synthetic with the 1-octyne.Compound 2b uses the method identical with 1c with above-mentioned synthetic 1b synthetic respectively with 2c.The preparation of side chain precursor (3b):

In the 500mL round-bottomed flask that contains 100mL acetone, add m-hydroxybenzaldehyde (5.00g, 40.94mmol), the 1-bromo-n-11 (9.65g, 41.02mmol) and K ₂CO ₃(8.48g, 61.36mmol).The reacting by heating mixture is to refluxing and reacting 10 hours.To react cooling and under vacuum, remove acetone.The resistates that obtains distributes between ether/water, the saturated NaHCO of organic layer ₃Twice of solution washing also used the salt water washing once.Organic layer MgSO ₄Drying leaches siccative then and removes under vacuum and desolvate, and purifying on silicagel column obtains 1.6876 yellow oil with 3%EtOAc/ hexane wash-out.Spectroscopic data is consistent with compound 3a, and this aldehyde prepares compound 3b by using with the same procedure for preparing the 1c description then.The preparation of side chain precursor (4c):

Except that compound 1a with the tert.-butyl acetate condensation before the not hydrogenation, use same procedure synthetic compound 4a with above-mentioned synthetic compound 1c.

In the 50ml round-bottomed flask, add 3-hydroxyl-3-(a dodecyne base benzyl) propionic acid tert-butyl ester 4a (346.8mg, 0.897) and be dissolved in 10ml MeOH/1mL and ice among the AcOH.With 10%Pd/C (304.5mg) standard hydrogenation 24h, by removing by filter catalyzer, under vacuum, obtain 302.4mg 3-(dodecylbenzyl) the propionic acid tert-butyl ester except that desolvating, remove the tert-butyl ester with the TFA processing then and obtain compound 4b.The preparation of side chain precursor (5a):

Under-78 ℃, in the 50ml round-bottomed flask that contains 5mL THF, add s-butyl lithium (1.56mmol, the cyclohexane solution of 1.2mL 1.3M).Be added dropwise to the isobutyl bromide tert-butyl ester (in-78 ℃ 2ml THF) in this mixture, stirring reaction 30min drips compound 1b (318.9mg, 1.16mmol in 2ml THF ,-78 ℃) then in reaction mixture.The reaction mixture 30min that obtains-78 ℃ of stirrings is warming up to 0 ℃ and stirred 1.75 hours in the above time of 30min then under this temperature, then with 2ml saturated aqueous ammonium chloride termination reaction and be warmed to room temperature.This reaction mixture between ether/water, distribute and organic layer with the salt water washing once and use MgSO ₄Dry.Leach siccative then and under vacuum, obtain the 370.5mg crude product, be racemic mixture except that desolvating.Remove this tertiary butyl ester with TFA then and obtain compound 5a.The preparation of side chain precursor (6a):

To containing the dense H of 100mL MeOH and 1mL ₂SO ₄The 250mL round-bottomed flask in add between the bromine anisic acid (5.00g, 21.6mmol).Heat resulting mixture to refluxing and keeping 24 hours.The cooling reaction solution also removes under vacuum and desolvates, and the solid that obtains is handled with ether, and organic layer washes twice with water, uses saturated NaHCO ₃Each washs once and uses MgSO the aqueous solution and salt solution ₄Dry.Leach siccative then and obtain the 4.72g crude product except that desolvating under vacuum, this crude product is without being further purified use.This product and 1-tetradecyne condensation and obtain similar alkene then with front embodiment through over hydrogenation.This methyl esters converts formic acid to through the following step: (538.4mg 1.48mmol) is suspended in the AcOH solution of 20ml 30%HBr and reflux, behind the backflow 24h this solution is poured in the 150ml water and with 2 * 200ml CH with methyl esters ₂Cl ₂Extraction.Organic phase is with a large amount of water washings and use MgSO ₄Drying leaches siccative then and obtain 398.7g crude product 6a except that desolvating under vacuum, and it is without being further purified use.The preparation of side chain precursor (7a):

Compound 6a (385.2mg) is added in the pyridine hydrochloride solid.Heat two solids to～220 ℃ of fusions and kept mixture reaction 3 hours, then with the reactant cooling and at CH ₂Cl ₂Distribute between/1N the HCl.Organic phase is washed 5 times with 1N HCl then and is used MgSO ₄Drying leaches siccative then and obtain 158.8mg crude product (7a) except that desolvating under vacuum, and it is without being further purified use.The preparation of side chain precursor (8b):

In the 250ml round-bottomed flask, be added in 60mL toluene/30mL 2 M Na ₂CO ₃Xenyl boric acid in the aqueous solution (2.00g, 10.1mmol) and Pd (PPh) ₄(980.0mg, 0.848mmol).In this soup compound, add 3-bromobenzaldehyde (in 10mL MeOH).The reacting by heating mixture is to refluxing and keeping reaction 20h.The cooling reactant, organic layer washes twice with water, and the salt water washing is once also with using MgSO ₄Drying leaches siccative then and removes under vacuum and desolvate.The solid that obtains is removed any residual 3-bromobenzaldehyde with cold hexane rinsing, stir this solid pulp then in hot hexane, and heat filtering is removed all solids.Filtrate is removed to desolvate under vacuum and is obtained purpose aldehyde 8a.The same procedure that is used in preparation 1c description changes into 8b with compound 8a and obtains inseparable diastereomer.The preparation of side chain precursor (9a):

With tert.-butyl acetate (2.02ml, anhydrous THF (25ml) solution 0.015mol) be chilled to-78 ℃ and drip n-Butyl Lithium (1.6M is in hexane) (9.35ml, 0.015mol).Drip behind the 45min methyl n-undecyl ketone (2.0g, THF solution 0.01mol) continue to stir at low temperatures 1 hour, then more than 15min with the reactant temperature to room temperature.Add excessive 1N HCl termination reaction, aqueous solution extracted with diethyl ether (2x), ether layer MgSO ₄Dry and under vacuum, concentrate and obtain crude product oily matter.Obtain 1.01g (yield 32%) tertiary butyl ester through silica gel chromatography (5% ethyl acetate/hexane).NMR is consistent with the purpose product.Handle the tert-butyl ester with trifluoroacetic acid then tert-butyl ester decomposition is quantitatively obtained compound 9a.The preparation of side chain precursor (10e): R=n-C ₅H ₁₁

At room temperature, to m-hydroxybenzaldehyde (1.00g, add in the THF solution (16mL) 8.20mmol) DEAD (1.29mL, 8.20mmol), PPh ₃(2.15g, 8.20mmol) and Pentyl alcohol (723mg, 8.20mmol).At room temperature stir the mixture and spend the night, with obtaining 1.02g (65%) purpose product 10a behind the silica gel purification.

Under 0 ℃, (6.70g adds Ph in THF solution 34.90mmol) to 10a ₃P=CHO (10.6g, 34.90mmol).At room temperature stir spend the night after, filter reaction mixture also concentrates under vacuum and obtains resistates, obtains 3.57g (47%) purpose unsaturated aldehyde 10b through the silica gel chromatography purifying.

With 10%Pd/C (1.64g, 1.55mmol) hydrogenation (1.5atm) 10b (3.37g, EtOAc solution (30mL) 15.5mmol).Stirring reaction liquid spends the night, through Celite plate filter reaction mixture.Concentrated filtrate and rinsing liquid under vacuum, resistates obtains 2.37g (70%) 10c through silica gel chromatography (10%EtOAc/ hexane).

Under 0 ℃, ((0.91mL 5.71mmol), adds TiCl subsequently to add the allyl group trimethyl silyl among the 1.26g, dichloromethane solution 5.71mmol) (23mL) to aldehyde 10c ₄(0.63mL, 5.71mmol).After stirring 1 hour under 0 ℃, with saturated NaHCO ₃The solution termination reaction, (75mL) dilutes this mixture with methylene dichloride, and organic layer is used saturated NaHCO successively ₃, water and salt water washing.The organic layer drying, concentrate and obtain 0.91g (61%) purpose allyl alcohol 10d through silica gel chromatography purifying (10%EtOAc/ hexane).

With allyl alcohol 10d (0.91g 3.47mmol) is dissolved in the aqueous acetone solution (each 7mL), and in this solution, add NMO (704mg, 5.21mmol) and OsO ₄(44mg, THF solution 0.17mmol).After at room temperature stirring 2 hours, use NaHSO ₃(750mg) termination reaction is to stop excessive oxygenant.(3 * 50mL) extract reaction mixture, the organic layer drying of merging and concentrated 850mg (82%) purpose three alcohol intermediates that obtain under vacuum through salt solution (10mL) dilution and with EtOAc.(850mg 2.87mmol) is dissolved in MeOH (30mL) and the water (6mL), at room temperature uses NaIO with the triol that so obtains ₄(1.38g 6.46mmol) handles this solution, and after 1 hour, reaction mixture filters through the Celite plate, and careful concentrated filtrate obtains corresponding beta-hydroxy aldehyde under vacuum.This aldehyde is dissolved in t-BuOH (14mL) and the hexanaphthene (2mL), in this solution, adds and contain KH ₂PO ₄(2.33g, 17.7mmol) and NaClO ₂(2.08g, (the 15mL H of water 23.0mmol) ₂O) solution, at room temperature stirring reaction is 5 hours, is acidified to pH=3 with 1N HCl then, and (3 * 50mL) extract the organic phase water of merging and salt water washing to reaction mixture with EtOAc.Organic layer drying and under vacuum, concentrating obtain crude acid 10e (1.5g ,～3.4mmol).

Also can making wherein with above-mentioned same procedure, R is n-C ₆H ₁₃, n-C ₇H ₁₅, n-C ₈H ₁₇, n-C ₉H ₁₉, n-C ₁₀H ₂₁And n-C ₁₄H ₂₉Compound.The preparation of side chain precursor (11d):

Under-78 ℃, ((10.9mL 68.69mmol), adds pure TiCl then to add the trimethylammonium allyl silicane among the 6.22g, dichloromethane solution 19.1mmol) (190mL) to chirality acetal 11a ₄(2.94mL, 26.71mmol).-78 ℃ of stirred reaction mixtures 1 hour, stirred 2 hours at-40 ℃ then, dilute with methyl alcohol (15mL) termination reaction and with methylene dichloride (200mL) this moment, the reaction mixture that obtains with 1N HCl (2 * 50mL), water and salt water washing, the organic layer drying also concentrates under vacuum and obtains resistates, and it obtains 5.51g (78%) purpose product 11b with silica gel chromatography (10%EtOAc/ hexane) purifying.11a's ¹H NMR (CDCl ₃): δ 4.73 (m, 1H), 4.21 (m, 1H), 3.86 (m, 1H), 1.75 (m, 1H), 1.60-1.10 (m, 35H), 0.80 (m, 3H).11b's ¹H NMR (CDCl ₃): δ 5.81 (m, 1H), 5.05 (m, 2H), 4.12 (m, 1H), 3.86 (m, 1H), 3.41 (m, 1H), 2.22 (m, 2H), 1.67-1.18 (m, 36H), 0.88 (m, 3H).

To 11b (8.56g, add in dichloromethane solution 23.3mmol) (155mL) PCC (10.0g, 46.5mmol).At room temperature stirring reaction is 18 hours, filters through the Celite plate then.Concentrated filtrate obtains the blush resistates under vacuum, obtains 8.36g (80%) methyl ketone intermediate (structure does not provide) through silica gel chromatography purifying (10%EtOAc/ hexane).With the intermediate that obtains here (8.36g 22.8mmol) is dissolved among THF (60mL) and the MeOH (30mL), adds 7.5M KOH (15mL) in this solution, at room temperature stir 3 hours after, remove partial solvent, remaining reaction mixture EtOAc/Et ₂O (ratio 3: 1,350mL) dilution.(3 * 50mL) and the salt water washing, the organic layer drying that obtains also concentrates under vacuum and obtains resistates the organic layer water, and it obtains 6.22g (96%) purpose product 11c through silica gel chromatography (10%EtOAc/ hexane) purifying, is white solid.11c's ¹H NMR (CDCl ₃): δ 5.75 (m, 1H), 5.06 (m, 2H), 3.56 (m, 1H), 2.23 (m, 1H), 2.07 (m, 1H), 1.75-1.17 (m, 28H), 0.80 (m, 3H).

(6.22g 22.0mmol) is dissolved in the THF aqueous solution (5.5mL water and 55mL THF), and (4.42g 33.0mmol), adds OsO subsequently to add NMO in this solution with methyl alcohol 11c ₄(280mg is dissolved among the THF, 1.10mmol).At room temperature stirring reaction spends the night, and this moment, adds sodium disulfide (4g), stirring reaction 2 hours, use EtOAc (300mL) dilution then, all (2 * 40mL) and the salt water washing, the organic layer drying that obtains also concentrates under vacuum and obtains corresponding three alcohol intermediates the mixture waters.It is dissolved in MeOH (200mL) and the water (40mL), in this solution, adds NaIO ₄(10.6g, 49.5mmol), at room temperature stir 1 hour after, obtain～10g the crude product beta-hydroxy aldehyde of (＞100%) through the Celite filter reaction mixture and with lacking silica gel chromatography (30%EtOAc/ hexane).This impure aldehyde that obtains is dissolved in t-BuOH (100mL) and the hexanaphthene (14mL), at room temperature in this solution, adds NaClO ₂(15.97g, 176mmol) and KH ₂PO ₄(17.8g, (50mL) aqueous solution 132mmol).At room temperature stirring reaction is 6 hours, arrives pH4 with 5N HCl termination reaction down at 0 ℃ then.EtOAc/Et with 3: 1 ₂The O mixed solvent (3 * 250ml) extractions, organic layer is with salt water washing, drying and concentrate the crude acid 11d that obtains 7.3g (＞100%), and it is directly used in coupled reaction.11d's ¹H NMR (CDCl ₃): δ 3.95 (m, 1H), 2.60-2.35 (m, 2H), 1.40-1.10 (m, 28H), 0.82 (m, 3H).The preparation of side chain precursor (12c):

In 1 liter round-bottomed flask, be added in tridecylic aldehyde among the anhydrous MeOH of 600ml (5.00g, 25.2mmol) and the hydrochloride of Gly-Ome (12.66g, 100.8mmol).In this reaction mixture, add NaCNBH ₃(1.787g, 28.44mmol) and at room temperature stirring reaction spends the night.Leach solid and remove under vacuum and desolvate, the gained resistates is at CH ₂Cl ₂/ saturated NaHCO ₃Distribute between the aqueous solution.Organic layer NaHCO ₃Solution washing 2 times is also used MgSO ₄Drying leaches siccative then and removes under vacuum and desolvate, and purifying on silicagel column obtains 2.94g white solid (12a) with 50%EtOAc/ hexane wash-out.

In the 50ml round-bottomed flask, be added in glycine derivative 12a among the anhydrous THF of 10ml (504.6mg, 1.86mmol), triethylamine (224.6mg, 2.22mmol).To wherein once adding (BOC) ₂O (494.3mg, 2.26mmol), stirring reaction 18 hours removes under vacuum then and desolvates, and gained oily matter is handled with EtOAc and is washed twice with 1N HCl, and MgSO is respectively washed once and used to water and salt solution ₄Drying leaches siccative then and obtain 463.3mg colorless oil (12b), its not purified use except that desolvating under vacuum.

(463.3mg 1.25mmol), to wherein adding 1.8ml 1N LiOH solution, stirs the reaction mixture that obtains and spends the night to add methyl esters 12b in the 50ml round-bottomed flask that contains 5ml THF.Add 1.8ml 1N HCl solution termination reaction, under vacuum, remove THF then and use CH ₂Cl ₂The water layer that extraction obtains 2 times.Organic phase MgSO ₄Drying leaches siccative then and obtain 246.2mg colorless oil (12c), its not purified use except that desolvating under vacuum.The preparation of side chain precursor (13a):

In the 50mL round-bottomed flask, the methyl esters 12b (499.7mg/1.84mmol) that obtains above is dissolved in 1: 1 acetic anhydride/pyridine mixtures, stirring reaction spends the night.Under vacuum, remove then and desolvate the oily matter CH that obtains ₂Cl ₂Handle and wash 2 times with 1N HCl, water and salt solution respectively wash 1 time and use MgSO ₄Drying leaches siccative then and obtain 319.9mg colorless oil (13a), its not purified use except that desolvating under vacuum.The general preparation of glycine side chain precursor (14-a):

For example following method is described with Fmoc-glycine-wang resin and the synthetic tridecanoyl-glycine of tridecanoic acid.

In the 100mL reaction tubes of a two ends glass end-sealing, (10g 4.4mmol) is added among 50mL 30% piperidines/DMF, jolts reaction 20 minutes and with DMF washing 3 times, uses washed with isopropyl alcohol 3 times, again with DMF washing 3 times with Fmoc-glycine-wang resin.In resin, add tridecanoic acid (4.708g, 50mL DMF solution 22mmol) and in this mixture, add HOBt (2.97g, 22mmol) and DIC (2.77g, 22mmol).Reactor placed on the vibrator spend the night.Use following solvent wash resin: DMF 3 times then, methylene dichloride 3 times, MeOH 3 times, THF 3 times and methylene dichloride 3 times.The dry resin pearl is 1 hour in vacuum oven, adds the TFA/H of 100 mL 95% in resin ₂O jolts reaction 1.5 hours and washs non--resin product and collection with TFA.Drying obtains white residue in vacuum oven, obtains tridecanoyl-glycine (1.14g, 95%) with the methylbenzene azeotropic distillation ¹H NMR (THF) 0.82-0.92 (t, J=7.2,3H), 1.2-1.4 (s, 18H), 1.52-1.62 (m, 2H), 2.10-2.17 (t, J=7.2,2H), 3.83-3.89 (d, J=7.1,2H), 7.04-7.17 (s, 1H), 10.8-10.9 (s, 1H).

Following structure I I is used for being described in the product that embodiment 1-17 obtains.

Embodiment 1 diastereomer 1-1 and 1-2's is synthetic:

With hydroxybenzotriazole (55.8mg, 0.413mmol) and 1-[3-(dimethylamino) propyl group]-(82.4mg 0.430mmol) is added in the 4ml dry DMF of compound 1c the 3-ethyl-carbodiimide hydrochloride.Stirred reaction mixture reaction at room temperature 10 hours, after this add Z-PSN (375.2mg, 0.271mmol).HPLC shows that the pseudobactin nuclear (Z-PSN) of CBZ-protection runs out of after 5 hours.Under vacuum, remove and desolvate and with 1: 1 ACN/H ₂O handles the resistates obtain and through preparation HPLC purifying, obtains 2 bigger peaks like this, and its mass spectrum shows that these peaks are corresponding to 2 diastereomer 1-1 (88.3mg) and 1-2 (166.3mg).Diastereomer 1-1 go the protection:

(82.0mg, 0.048mmol), degassing back is to reaction mixture adding 89.1mg 10%Pd/C and at 1atm H to add 10ml MeOH/1ml ice AcOH and diastereomer 1-1 in a 50ml round-bottomed flask ₂Hydrogenation 30 minutes.Remove catalyzer after filtration after preparation HPLC purifying and lyophilize obtain 21.7mg compound 1-3 (R ¹=H).MS (Ionspray) is for C ₅₈H ₉₄ClN ₁₂O ₁₉(M+H) ⁺Calculated value is 1297.64, and experimental value is 1297.8.Diastereomer 1-2 go the protection:

As described in for diastereomer 1-1 with the 10%Pd/C hydrogenation diastereomer 1-2 (152.8mg of 152.8mg, 0.089mmol) 30 minutes, HPLC shows the raw material completely consumed and forms two product peaks, obtains compound 1-4 (18.0mg) and compound 1-5 (11.3mg) after preparation HPLC purifying and lyophilize.Compound 1-4:MS (Ionspray) is for C ₅₈H ₉₄ClN ₁₂O ₁₉(M+H) ⁺Calculated value is 1297.89, and experimental value is 1297.8.Compound 1-5:MS (Ionspray) C ₅₈H ₉₂ClN ₁₂O ₁₈(M+H) ⁺Calculated value is 1279.63, and experimental value is 1281.7.

Listed compound is with acidylate same as described above and goes guard method in the following table 1, with the side chain precursor synthetic that marks.For each compound of listing in the table 1, R ¹Be hydrogen.

Table 1

Though * the compound of this compound and no N-methyl shows very low activity, wherein alkyl chain is increased to the compound of C15 continuously from C11, its active obviously rising.This class side chain can be by preparation method's preparation of describing in the preparation of 14-a.

Embodiment 15 compound 15-1's is synthetic:

In the 50ml round-bottomed flask, add 31.3mg (0.0237mmol) compound 12-1 and 5ml TFA (being chilled to 0 ℃ in advance).Under this temperature, stir this reaction mixture 15min, under vacuum, remove TFA, obtain 24.3mg compound 15-1 with water treatment resistates and lyophilize then.Do not need to be further purified.

The preparation of connection compound 16a, 16b, 16c and the 16d of the side chain that embodiment 16 embodiment 16 explanation beta-aminos replace:

Heating tert-butoxycarbonyl methylene tri phenyl phosphorane (5.0g, 13.3mmol) and lauric aldehyde (2.2ml, toluene 10mmol) (50ml) solution backflow 1 hour 20 minutes.Filter this solution to remove dephosphorization reagent through silica gel plug, concentrate under vacuum then and obtain crude product oily matter, this oily matter is through the silicagel column purifying, and the hexane solution wash-out of the ethyl acetate with 2% obtains the compound 16a of 2.36g (yield 84%).MS-283.4 (M+1) NMR is consistent with structure.

Under-78 ℃, (1.19ml, (0.42ml in THF 2.0mmol) (5ml) solution, drips 16a (500mg, THF solution 1.77mmol) then 1.9mmol) slowly to be added to (R)-benzyl methyl-benzyl amine with butyllithium (1.6M is in hexane).Under-78 ℃, stirred the mixture 1 hour, and then reaction mixture was poured into saturated NH ₄Also use extracted with diethyl ether (2x) in the Cl solution.Diethyl ether solution MgSO ₄Dry and under vacuum, concentrate and obtain 0.95g16b, be crude product oily matter, this oily matter is used for next step without purifying.

Under 55 ℃, with ethanol (25ml) solution and the Pd (OH) of 16b (0.95g) ₂/ C (0.47g) places 60psi H ₂Following 18 hours.Filtering suspension liquid concentrates under vacuum then and obtains 280mg (yield 53%) crude product 16c, and it is directly used in next step.

Mixing cpd 16c in THF (10ml) (280mg, 0.93mmol) (274mg, 1.1mmol) also spend the night with N-benzyloxycarbonyloxy succinimide by stirring.Under vacuum, remove desolvate and with the oily product through silica gel chromatography, obtain the tertiary butyl ester 16d of 268mg (yield 66%) as eluent with the hexane solution of 5% ethyl acetate.NMR and object construction coincide.Under 0 ℃, (97mg 0.224mmol) is dissolved in the trifluoroacetic acid (2ml) 0.5 hour and removes tertiary butyl ester (16d) by quantitative yield with this ester.Compound 16-1's is synthetic:

Compound 16d is dissolved among the DMF (2ml), add hydroxybenzotriazole (36.4mg, 0.269mmol) and 1-[3-(dimethylamino) propyl group]-the 3-ethyl-carbodiimide hydrochloride (47.1mg, 0.246mmol) and stirred this solution 18 hours.Add the CBZ protection pseudobactin nuclear (Z-PSN) (277mg, 0.185mmol) and restir reaction 18 hours.Reaction product obtains the pseudobactin derivative of the CBZ protection of 136.3mg (yield 42%) acidylate through HPLC purifying and lyophilize, is white solid.MS-1743 (M) and NMR are consistent with predetermined structure.

(130mg, methyl alcohol 0.0746mmol) (10ml) and acetate (1.5ml) solution and 10%Pd/C (120mg) placed the hydrogen capsule following 20 minutes with the compound of acidylate.Filter this solution and use preparation HPLC purifying, lyophilize obtains the trifluoroacetate of 23mg (yield 18%) 16-1, is white solid.MS-1206.8 (M) and NMR are consistent with predetermined structure.Compound 16-1 anti-candida albicans and the very low or non-activity of novel Cryptococcus activity are compared with hydroxy analogs, its active significantly reduction.

The preparation of the connection side chain precursor 17d of embodiment 17 embodiment 17 explanation chiral side chains:

(240mg adds 4A molecular sieve (4g) in THF solution 0.84mmol), add pure Ti (OPr-I) then to (S)-dinaphthol ₄(0.25mL, 0.84mmol).Reaction mixture reddens very soon and keeps red, and the chiral catalyst (17b) for preparing thus is used for following reaction.

Under-78 ℃, more than 15 minutes, in the THF solution (4mL) of (S)-dinaphthol-Ti catalyzer 17b (0.42mmol) of new system, add and contain trimethyl silyl dimethyl ketene acetal (trimethylsilyldimethylketene acetal) (0.43mL, 2.1mmol) and unsaturated aldehyde 17a (500mg, THF solution 2.1mmol).-78 ℃ of following stirring reactions 1 hour, at room temperature stir then and spend the night.Use saturated NaHCO ₃The solution termination reaction is also used EtOAc (100mL) extraction.Organic layer NaHCO ₃, salt water washing and use anhydrous MgSO ₄Drying is filtered and is concentrated under vacuum, and resistates obtains 257mg (36%) purpose product 17c with silica gel chromatography (10%EtOAc/ hexane).17c's ¹H NMR (CDCl ₃): δ 5.29 (m, 2H), 3.65 (s, 3H), 3.53 (t, J=7.3Hz, 1H), 2.33 (d, J=6.3Hz, 1H, 3 '-OH), 1.97 (m, 4H), 1.65-1.22 (m, 20H), 1.13 (s, 3H), 1.12 (s, 3H), 0.84 (m, 3H).

((259mg, THF solution (5mL) 0.76mmol) spends the night 50 ℃ of reacting by heating 1.52mmol) to handle 17c for 0.30mL, 5N with the NaOH aqueous solution.Reaction mixture is chilled to 0 ℃ and be acidified to pH=3 with 5N HCl, uses EtOAc (75mL) extraction then, organic layer water and salt water washing, dry and concentrate under vacuum and obtain 222mg (90%) crude acid 17d, it is directly used in side chain coupling and reacts.17d's ¹H NMR (CDCl ₃): δ 5.28 (m, 2H), 3.56 (m, 1H), 1.95 (m, 4H), 1.60-1.10 (m, 26H), 0.83 (m, 3H).The preparation of 17-1 and 17-2:

At room temperature, with HOBt (90.5mg, 0.67mmol) and EDCI (128mg, 0.67mmol) processing crude acid 17d (240mg, THF solution (7mL) 0.74mmol).Stir add after 1.5 hours DMAP (41mg, 0.33mmol).Restir is added in the pseudobactin nuclear of the CBZ-protection among the DMF (4mL) after 2 hours (614mg, 0.44mmol) and at room temperature stirring reaction spends the night.Reaction mixture is through preparation HPLC purifying, and lyophilize obtains (160mg, 21%) purpose product.

Under 0 ℃, (77mg 0.38mmol) handles compound (53.4mg, acetonitrile solution 0.032mmol) (2mL) that CBZ-protects with TMSI.After 100 minutes, with 1: 1CH ₃CN/H ₂The O termination reaction, the reaction mixture that obtains obtains 25.8mg (65%) purpose final product 17-1 through anti-phase preparation HPLC purifying.

Use R ^*For the suitable raw material of hydrogen makes compound 17-2 with above-mentioned identical method.

The preparation of the connection side chain precursor 18d of embodiment 18 embodiment 18 explanation chirality thiazolinyl side chains:

Under-78 ℃, to 18a (993mg, 3.73mmol) dichloromethane solution (25mL) in add dimethyl ketene contract methyl alcohol contract trimethylammonium first silanol (methyltrimethylsilyl dimethylketene acetal) (2.27mL, 11.2mmol) and pure TiCl ₄(0.49mL, 4.48mmol).After 2 hours, use MeOH (5mL) termination reaction down at-78 ℃.(3 * 40mL) extract the organic layer NaHCO of merging to reaction mixture with methylene dichloride ₃With the salt water washing.The organic layer drying also concentrates under vacuum and obtains resistates, and this resistates obtains 837mg (61%) purpose product 18b through chromatogram purification (15-20%EtOAc/ hexane).18a's ¹H NMR (CDCl ₃): δ 6.28-5.87 (m, 2H), 5.62-5.43 (m, 2H), 4.75 (m, 1H), 4.22 (m, 1H), 3.87 (m, 1H), 2.08-1.93 (m, 2H), 1.80-1.60 (m, 3H), 1.60-1.40 (m, 3H), 1.38-1.10 (m, 15H) .18b ¹H NMR (CDCl ₃): 5.98-5.90 (m, 2H), 5.55-5.47 (m, 2H), 4.08 (m, 1H), 3.86 (m, 1H), 3.61 (s, 3H), 3.50 (m, 1H), 2.00-1.96 (m, 2H), 1.74-1.63 (m, 3H), 1.50-1.00 (m, 24H).

To 18b (837mg, add in dichloromethane solution 2.27mmol) (22mL) PCC (0.98g, 4.54mmol).At room temperature stirring reaction is 18 hours, filters through the Celite plate then, and concentrated filtrate obtains pink resistates under vacuum, obtains 441mg (53%) purpose methyl ketone 18c through silica gel chromatography purifying (20%EtOAc/ hexane).18c's ¹H NMR (CDCl ₃): δ 5.92-5.87 (m, 2H), 5.48-5.43 (m, 2H), 3.88 (m, 1H), 3.56 (s, 3H), 3.49 (m, 1H), 2.59 (dd, J=6.4,14.7Hz, 1H), 2.29 (dd, J=5.9,15.2Hz, 1H), 2.06 (s, 3H), 1.96 (m, 2H), 1.63 (m, 2H), 1.40-0.90 (m, 20H).

At room temperature, (440mg, THF 1.20mmol) (4mL) and methane (methane) (2mL) add 7.5M KOH (1mL) in the solution to 18c.After at room temperature stirring 4 hours, be acidified to pH=3 with 1N HCl, with EtOAc (3 * 30mL) extractive reaction mixtures, the extraction liquid water that merges (2 * 10mL) and the salt water washing, organic layer drying and concentrate under vacuum and obtain 388mg (＞100%) crude acid 18d, it is directly used in the linked reaction of side chain.The preparation of compound 18-1:

To crude acid 18d (add in～1.66mmol) the THF solution (10mL) HOBt (244mg, 1.66mmol) and EDCI (318mg, 1.66mmol).After at room temperature stirring 5 hours, add the Alloc-protection pseudobactin nuclear (614mg, DMF solution (5mL) 0.50mmol), at room temperature stir several days after, to reaction mixture add DMAP (61mg, 0.50mmol).Behind the restir 12 hours, reaction mixture is through the reversed-phase HPLC purifying, and lyophilize obtains the acylated derivatives of 225mg (30%) alloc-protection then.

At room temperature, (240mg adds PdCl in THF 0.16mmol) (20mL) and HOAc (1mL) solution to the acylated derivatives of protecting to the alloc-of the degassing ₂(PPh ₃) ₂(23mg, 0.032mmol) and Bu ₃SnH (0.87mL, 3.23mmol).1.5 after hour, reaction mixture obtains 33mg (17%) compound 18-1 through preparation HPLC purifying.

The compound of listing in the foregoing description, also prepared following N-acyl derivative, these derivatives show limited active or non-significant activity.Even these compounds can not be used as mycocide, they also can provide valuable reference for the compound that design has an optimum activity.

Except as otherwise noted, each compound among the embodiment all shows the activity of suitable anti-candida albicans, novel Cryptococcus, Aspergillus fumigatus, Candida parapsilosis or Histoplasma capsulatum.But having observed according to these compounds of synthetic has following basic trend in activity.The stereochemistry of beta-hydroxy is preferably the R type; Do not consider stereochemistry and degree of unsaturation, long alkyl chain length (that is C, ₁₂-C ₂₀) be tending towards relatively short alkyl chain (for example,＜C ₁₁) have a higher activity; Remove beta-hydroxy, α, α-two replaces, is reduced in alkyl chain length, the special rigidity in alkyl and the alkoxy substituent and increase side chain in the side chain and is tending towards having the low activity of flexible chain than long.

Therefore, treatment is preferred to the alkyl group side chain represented of following structure to fungicidal:

Wherein

R ^aAnd R ^{A '}Independent is hydrogen or methyl or R ^aOr R ^{A '}One of be alkylamino and R ^bOr R ^{B '}Form hexavalent cycloalkyl ring, hexavalent aromatic ring or two key or and R together ^cForm the hexavalent aromatic ring together;

R ^bOr R ^{B '}Independent is hydrogen or methyl, and R ^bOr R ^{B '}Be hydroxyl, condition is to work as R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydrogen and R ^fBe n-hexyl, n-octyl or positive decyl, perhaps R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydroxyl and R ^fDuring for n-octyl, n-nonyl or positive decyl, R ^{B '}It is not hydroxyl;

R ^cBe hydrogen, hydroxyl, C ₁-C ₄Alkoxyl group, hydroxy alkoxy base are perhaps with R ^eForm 6 yuan of aromatic rings or C together ₅-C ₆Cycloalkyl ring;

R ^eBe hydrogen, or and R ^fBe hexa-atomic aromatic ring, C together ₅-C ₁₄Hexa-atomic aromatic ring or C that alkoxyl group replaces ₅-C ₁₄The hexa-atomic aromatic ring that alkyl replaces;

R ^fBe C ₈-C ₁₄Alkyl, or C ₅-C ₁₁Alkoxyl group.

Claims (15)

1. compound and its pharmacologically acceptable salt and its solvate of representing of a structure I

Wherein R is

Wherein

R ^aAnd R ^{A '}Independent is hydrogen or methyl or R ^aOr R ^{A '}Be alkylamino and R ^bOr R ^{B '}Form hexa-atomic cycloalkyl ring, hexa-atomic aromatic nucleus or two key or and R together ^cForm hexa-atomic aromatic nucleus together;

R ^bAnd R ^{B '}Independent is hydrogen, halogen or methyl, perhaps R ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl, condition is to work as R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydrogen and R ^fBe n-hexyl, n-octyl or positive decyl, or R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydroxyl and R ^fDuring for n-octyl, n-nonyl or positive decyl, R ^bIt is not hydroxyl;

R ^cBe hydrogen, hydroxyl, C ₁-C ₄Alkoxyl group, hydroxy alkoxy base, or and R ^eForm hexa-atomic aromatic nucleus or C together ₅-C ₆Cycloalkyl ring;

R ^eBe hydrogen or and R ^fForm hexa-atomic aromatic nucleus, C together ₅-C ₁₄Hexa-atomic aromatic nucleus or C that alkoxyl group replaces ₅-C ₁₄The hexa-atomic aromatic nucleus that alkyl replaces and

R ^fBe C ₈-C ₁₈Alkyl, C ₅-C ₁₁Alkoxyl group or xenyl; Or R is

Wherein

R ^gBe hydrogen or C ₁-C ₁₃Alkyl and

R ^hBe C ₁-C ₁₅Alkyl, C ₄-C ₁₅Alkoxyl group, (C ₁-C ₁₀Alkyl) phenyl ,-(CH ₂) _nAryl or-(CH ₂) _n-(C ₅-C ₆Cycloalkyl), n=1-2 wherein; Or R is Wherein

R ⁱBe hydrogen, halogen or C ₅-C ₈Alkoxyl group and

M is 1,2 or 3; R is Wherein

R ^jBe C ₅-C ₁₄Alkoxyl group or C ₅-C ₁₄Alkyl and p=0,1 or 2; R is Wherein

R ^kBe C ₅-C ₁₄Alkoxyl group; Or R is-(CH ₂)-NR ^m-(C ₁₃-C ₁₈Alkyl), R wherein ^mFor H ,-CH ₃Or-C (O) CH ₃

2. the compound of claim 1, wherein structure I has following stereochemistry:

3. the compound of claim 1, wherein R is

Wherein

R ^aAnd R ^{A '}Independent is hydrogen or methyl, perhaps R ^aOr R ^{A '}Be alkylamino, with R ^bOr R ^{B '}Form hexa-atomic cycloalkyl ring, hexa-atomic aromatic nucleus or two key together, or and R ^cForm hexa-atomic aromatic nucleus together;

R ^bAnd R ^{B '}Independent is hydrogen, halogen or methyl or R ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl, condition is: work as R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydrogen and R ^fBe n-hexyl, n-octyl or positive decyl, or R ^a, R ^b, R ^d, R ^eBe hydrogen, R ^cBe hydroxyl and R ^fDuring for n-octyl, n-nonyl or positive decyl, R ^{B '}It is not hydroxyl;

R ^cBe hydrogen, hydroxyl, C ₁-C ₄Alkoxyl group, hydroxy alkoxy base or and R ^eForm hexa-atomic aromatic nucleus or C together ₅-C ₆Cycloalkyl ring;

R ^eBe hydrogen, or and R ^fBe hexa-atomic aromatic nucleus, C together ₅-C ₁₄Hexa-atomic aromatic nucleus or C that alkoxyl group replaces ₅-C ₁₄The hexa-atomic aromatic nucleus that alkyl replaces; With

R ^fBe C ₈-C ₁₈Alkyl, C ₅-C ₁₁Alkoxyl group or xenyl.

4. the compound of claim 3, wherein R ^{B '}Be hydroxyl, condition is: work as R ^a, R ^b, R ^d, R ^eBe hydrogen and R ^fDuring for n-hexyl, n-octyl or positive decyl, R ^cBe not hydrogen or work as R ^fR during for n-octyl, n-nonyl or positive decyl ^cIt is not hydroxyl.

5. aforementioned claim in each claimed compound be used for the treatment of purposes in the medicine of whole body fungi infestation or fungal skin infections in preparation.

6. pharmaceutical composition, it comprises pseudobactin compound and pharmaceutically acceptable carrier of claim 2.

7. the method for a treatment anti-fungal infection in the animal of needs treatments, it comprises the step to the pseudobactin compound in the described animals administer claim 2.

8. the method for preparing pseudobactin nuclear, it comprises provides pseudobactin compound and described pseudobactin compound with the N-acyl group alkyl group side chain that comprises at least one γ or δ hydroxyl to prepare the step that described pseudobactin is examined with a kind of acid-respons.

9. the method for claim 8, wherein said pseudobactin nuclear is represented by structure I-A.

Wherein R ' is-NH ₂Or-NH _p-P _g, P wherein _gFor amino protecting group and p are 0 or 1.

10. the method for claim 8, wherein said pseudobactin compound with the N-acyl group alkyl group side chain that comprises at least one γ or δ hydroxyl is selected from pseudobactin A, pseudobactin A ' and pseudobactin C.

11. the method for claim 8, wherein said acid are trifluoroacetic acid and acetate.

12. the method for claim 11, wherein said acid are trifluoroacetic acid.

13. pseudobactin nuclear, it is by the method preparation of claim 8.

14. the pseudobactin of claim 13 nuclear, wherein said nuclear is represented by structure I-A. Wherein R ' is-NH ₂Or-NH _p-P _g, P wherein _gFor amino protecting group and p are 0 or 1.

15. examine by the pseudobactin that structure I-A represents.

Wherein R ' is-NH ₂Or-NH _p-P _g, P wherein _gFor amino protecting group and p are 0 or 1.