CN1373770A - Pseudomycin prodrugs - Google Patents

Abstract

A pseudomycin prodrug represented by structure (A) where R1 is an acyloxyalkylcarbamate linkage is described. The prodrug demonstrates antifungal activity with less adverse side effects.

Description

Pseudomycin prodrugs

Invention field

The present invention relates to pseudobactin (pseudomycin) compound, especially pseudobactin compound prodrug.

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-DabL-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, " FEBSLetters, 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 a kind of represented Pseudomycin prodrugs, its pharmaceutically useful salt and solvate thereof of following structural formula as anti-mycotic agent,

Wherein R is Wherein

R ^aAnd R ^{A '}Be H 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 H, halogen or methyl, perhaps R independently ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl;

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

R ^eBe H, 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; R is

Wherein

R ^gBe H 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 H, 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 H ,-CH ₃Or-C (O) CH ₃R ¹Be H, acyloxy methylene radical-1 independently, 3-dioxole-2-ketone (compound 1 for example described below (a)) or acyloxy methylene radical carboxylicesters (for example following compound 1 (b))

Wherein

R ^1aBe H, C ₁-C ₁₀Alkyl, C ₁-C ₁₀Thiazolinyl, benzyl or aryl, and

R ^1bBe H or methyl

Condition is at least one R ¹Be acyloxy methylene radical-1,3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters; R ²And R ³Be independently-OR ^2a, or-N (R ^2b) (R ^2c),

Wherein

R ^2aAnd R ^2bBe H, C independently ₁-C ₁₀Alkyl (for example methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, sec-butyl, the tertiary butyl etc.), C ₃-C ₆Cycloalkyl (for example cyclopropyl, cyclobutyl, cyclopentyl, cyclopentyl methylene radical, methylcyclopentyl, cyclohexyl etc.), hydroxyl (C ₁-C ₁₀) alkyl, alkoxyl group (C ₁-C ₁₀) alkyl (for example methoxy ethyl) or C ₂-C ₁₀Thiazolinyl, amino (C ₁-C ₁₀) alkyl ,-or two-alkylamino (C ₁-C ₁₀) alkyl, aryl (C ₁-C ₁₀) alkyl (for example benzyl), heteroaryl (C ₁-C ₁₀) alkyl (for example 3-pyridylmethyl, 4-pyridylmethyl) or the assorted alkyl (C of ring ₁-C ₁₀) alkyl (for example N-tetrahydrochysene-1,4-oxazinyl ethyl and N-piperazinyl ethyl), perhaps

R ^2bBe aminoacid alkyl ester the alkyl carboxylates residue (for example-CH ₂CO ₂CH ₃,-CH (CO ₂CH ₃) CH (CH ₃) ₂,-CH (CO ₂CH ₃) CH (phenyl) ,-CH (CO ₂CH ₃) CH ₂OH ,-CH (CO ₂CH ₃) CH ₂(p-hydroxybenzene) ,-CH (CO ₂CH ₃) CH ₂SH ,-CH (CO ₂CH ₃) CH ₂(CH ₂) ₃NH ₂,-CH (CO ₂CH ₃) CH ₂(4-or 5-imidazoles) ,-CH (CO ₂CH ₃) CH ₂CO ₂CH ₃,-CH (CO ₂CH ₃) CH ₂CO ₂NH ₂Deng), and

R ^2cBe H or C ₁-C ₆Alkyl.

In another embodiment of the present invention, a kind of medicinal preparations is provided, it comprises above-mentioned Pseudomycin prodrugs and pharmaceutically useful carrier.

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 the above-mentioned Pseudomycin prodrugs of described animals administer.The present invention also provides the purposes of above-mentioned Pseudomycin prodrugs medicine of anti-fungal infection in production is used for the treatment of animal.

Definition

Except as otherwise noted, term used herein " 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] heptane 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 be to be substituted or unsubstituted.Alkenyl part in alkenyloxy, alkenoyl or the chain acid ester has top identical definition.

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.

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 " ad hoc hypothesis and allowing is included on the alkyl of this area routine and replaces, 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 " prodrug " refers to a class medicine, and they produce pharmacological action because metabolic process transforms (being bio-transformation) in vivo.In the present invention, the Pseudomycin prodrugs compound contains linker, and described linker can split through esterase in blood plasma, thereby produces described active drug.

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 side effect of prodrug derivant that the applicant finds the natural or semi-synthetic product of pseudobactin is than the low of corresponding natural product and kept the effect of anti-candida albicans (C.albican), novel Cryptococcus (C.neoformas) and Aspergillus fumigatus (A.fumigatus) in vivo.This prodrug is by producing to form acyl substituent with at least one acidylate in the side amino that Methionin in the pseudobactin cyclic peptide member ring systems or 2,4-diamino-butanoic peptide unit link to each other.Acylating agent (or linker) normally contains the acyloxy methylene radical-1 of suitable leavings group; 3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters acylated compounds are so that can form and the amino amino-formate bond that is connected of the structural side of pseudobactin.Suitable leavings group to those skilled in the art for known and comprise for example group of p-nitrophenyl oxygen base and N-oxygen base succinimide.

Acyloxy methylene radical-1,3-dioxole-2-keto acyl compound can use the synthesis path shown in the following route I synthetic.For illustrative purposes, a specific acylated compounds has been described.Yet, it will be understood by those skilled in the art that people can use identical basic synthetic method to synthesize various derivatives.

Route I is for these synthesis step more detailed descriptions, referring to the preparation part of following embodiment.

Acyloxy methylene radical carboxylicesters acylated compounds can use the synthesis path shown in the following route II synthetic.For illustrative purposes, a specific acylated compounds has been described.Yet, it will be understood by those skilled in the art that people can use identical basic synthetic method to synthesize various derivatives.

Route II is for these synthesis step more detailed descriptions, referring to the preparation part of following embodiment.

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 Pseudomonas Syringae) " 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 NaturalProducts) " that people such as Kulanthaivel submitted on April 14th, 2000, its sequence number is PCT/US00/08727, and US5,576,298 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 MSU16H 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 from Pseudomonassyringae possessing broad-spectrum antifungal activity; " J.Gen.Microbiology, 137,2857-2865 (1991); And people such as Lamb " Transposonmutagenesis and tagging of fluorescent pseudomonas:Antimycoticproduction is necessary for 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 MSU174, 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 is ATCC 67028.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

67 H1 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 MSU16H, 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.

In addition, this prodrug can be by the semi-synthetic compound formation of N-acyl group.Semi-synthetic pseudobactin compound can synthesize by the N-carboxyl groups on the exchange L-Serine unit.The example of various N-acyl derivatives be described in people such as Belvo in the PCT patent application serial number that is entitled as " Pseudomycin N-acyl side-chain analogs (Pseudomycin N-Acyl Side-Chain Analogs) " of the application same period _ _ _ _ in, be introduced into this paper as a reference.In general, using four following synthesis steps is its semi-synthetic compound with the production of naturally occurring pseudobactin compound: (1) selectivity amido protecting; (2) chemistry of described N-acyl side-chain or enzymatic deacylation; (3) with different side chains acidylate again; (4) described amino goes protection.

Position 2,4 and 5 side amino can use the arbitrary standards method known to the skilled in amido protecting field to be protected.The accurate kind of used amido protecting group is not crucial; as long as the amino of derivatize ensuing relatively stable reaction conditions and described blocking group on other position of intermediate molecule can optionally be removed suitable the time, and to the molecule remainder that comprises other amido protecting group arbitrarily not destruction get final product.The amido protecting group that is fit to comprises benzyloxycarbonyl, to the nitro benzyloxycarbonyl, to the bromo-benzyloxy-carbonyl, to methoxyl group benzyloxy base carbonyl, p-methoxyphenyl azo benzyloxycarbonyl, to phenylazo benzyloxycarbonyl, tert-butoxycarbonyl, cyclopentyloxy carbonyl and phthaloyl imino.Preferred amido protecting group is tert-butoxycarbonyl (t-Boc), allyloxy carbonyl (Alloc), phthaloyl imino and benzyloxycarbonyl (CbZ or CBZ).The further example of the blocking group that is fit to is described in T.W.Greene, " Protective Groups in Organic Synthesis, " JohnWiley and Sons, New York, N.Y., (2nd ed., 1991), at chapter 7.

Deacylation with N-acyl group of γ or δ hydroxylation side chain (for example 3,4-dihydroxyl ten tetra-carbonic esters) can realize with the pseudobactin compound of the acid treatment amido protecting in the water-containing solvent.The acid that is fit to comprises acetate and trifluoroacetic acid.Preferred acid is trifluoroacetic acid.If the use trifluoroacetic acid can be in room temperature or near carrying out this reaction under the room temperature.Yet when using acetate, reaction is carried out under about 40 ℃ usually.The aqueous solvent system that is fit to comprise acetonitrile, water, and composition thereof.Organic solvent strengthens this speed of response; Yet, add organic solvent and may cause other by product.The pseudobactin compound (for example pseudobactin B and C ') that lacks δ or γ hydroxyl on side chain can be through the enzymatic deacylated tRNA.The deacylase that is fit to comprises polymyxin acyltransferase (164-16081 acyl transferring enzyme (crude product) or 161-16091 acyl transferring enzyme (pure product) can be from Wako Pure Chemical Industries, and Ltd. obtains) or ECB deacylase.Can use the standard deacylation step of well known to a person skilled in the art to carry out this enzymatic deacylation.For example, use the conventional steps of polymyxin acyltransferase in following document, to find: Yasuda, N. etc., Agric.Biol.Chem., 53,3245 (1989) and Kimura, Y. etc., Agric.Biol.Chem., 53,497 (1989).

The respective acids of deacylated tRNA product (being also referred to as Pseudomycin nuclear) required acyl group of use under the situation that the carbonyl activator is arranged is acidylate again." carbonyl activating group " is meant the carbonyl substituted base that promotes nucleophilic addition at this carbonyl place.The activation substituting group that is fit to is to have those of clean electrophilic effect on described 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 methanesulfonate ester, ethylsulfonic acid ester group, benzene sulfonate and p-toluenesulfonic esters); And halogenide (for example muriate, bromide and iodide).

Various acid can be used for this process for acylating.The acid that is fit to comprises the lipid acid that contains one or more side aryl, alkyl, amino (comprise primary, the second month in a season and tertiary amine), hydroxyl, alkoxyl group and amido; Contain the lipid acid of nitrogen or oxygen at aliphatic chain; Aromatic acid with alkyl, hydroxyl, alkoxyl group and/or alkylamino replacement; With the heteroaromatic acid that replaces with alkyl, hydroxyl, alkoxyl group and/or alkylamino.

Perhaps, can use solid-phase synthesis, wherein with the coupler of hydroxybenzotriazole resin (HOBt-resin) as acylation reaction.

In case amino deacylated tRNA and again acidylate (as mentioned above) can under the situation that hydrogenation catalyst (for example 10%Pd/C) arranged, remove amido protecting group (2,4 and 5) afterwards by hydrogenation.When the amido protecting group is allyloxy carbonyl, can use tributyltin hydride and dichloride triphenylphosphine palladium to remove described blocking group.This specific protection/go the advantage of protection scheme is to have reduced the unitary vinyl hydrogenation of Z-Dhb potential in this pseudobactin structure.

Then by following production prodrug: acidylate Methionin or with side amino that 2,4-diamino-butanoic peptide unit in the semi-synthetic pseudobactin compound of N-acyl group modification links to each other at least one, to form the requisite carbamate key.

Side hydroxy-acid group by aspartic acid and/or hydroxyl aspartic acid units in amidation or the esterification pseudomonas prime ring can synthesize the prodrug pseudobactin compound of other modification.The example of the derivative of various sour modifications is described in be entitled as " the Pseudomycin Amide ﹠amp of people such as Chen in the application same period; Ester Analogs " the PCT patent application serial number _ _ _ _ in, and be introduced into this paper as a reference.The derivative of these sour modifications can be by with aforesaid any prodrug and suitable alcohol or the amine condensation produces corresponding ester or acid amides forms.

The formation of these ester groups can use the standard esterif iotacation step of well known to a person skilled in the art to realize.Esterification under acidic conditions typically is included in and under the situation of protonic acid (for example HCl, TFA etc.) described pseudobactin compound is dissolved in or is suspended in the suitable alcohol.Under alkaline condition, described pseudobactin compound reacts with the alkylogen that suits usually under the situation that weak base (for example sodium bicarbonate and salt of wormwood) is arranged.

The formation of amide group can be used and well known to a person skilled in the art that the standard amide step realizes.Yet the selection of coupler provides the selective modification of described acid groups.For example, use benzotriazole-1-base oxygen base-tripyrrole Wan Ji Phosphonium hexafluorophosphate (PyBOP), make people can separate pure monoamide and (under some situation) pure diamide on 8 of residues simultaneously as coupler.Yet, use adjacent benzotriazole-1-base-N, N, N ', N '-tetramethyl-urea a tetrafluoro borate (TBTU) forms monoamide on 3 of the residues as coupler deflection.

This Pseudomycin prodrugs can be isolated and use with itself or with its pharmacologically acceptable salt or solvate forms.As mentioned above, this prodrug is by forming at least one acyloxy alkyl carbamate key preparation.Term " pharmacologically acceptable 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.

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

Typically activeconstituents (being Pseudomycin prodrugs) 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.Goods 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 physical characteristics of animal, the severity of animal illness, the mode that is used for administration and animal species.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 spraying), oral, injection and sucks.Used concrete treatment process will depend on the type of injecting.

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, Pseudallescheriaboydii, posadasis spheriforme, sporotrichum schenckii etc.

So compound of the present invention and preparation are useful at the medicine that preparation is used for resisting systemic fungal infection or fungal skin infections.Therefore, provide a kind of method that suppresses fungi activity, this method comprises Pseudomycin prodrugs of the present invention is contacted with fungi.Preferred method comprises inhibition Candida albicans or activity of Aspergillus fumigatus.Term " contact " comprises that The compounds of this invention combines or engages with fungi, and perhaps the surface contacts or is 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 parasite and 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 normally comprises following animal: 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

The listed separately material of abbreviation representative below in whole embodiment, using:

The ACN-acetonitrile

The TFA-trifluoroacetic acid

The DMF-dimethyl formamide

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

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

The CBZ=benzyloxycarbonyl, C ₆H ₅CH ₂-O-C (O)-

PyBOP=benzotriazole-1-base oxygen base-tripyrrole Wan Ji Phosphonium hexafluorophosphate

The adjacent benzotriazole of TBTU=-1-base-N, N, N ', N '-tetramethyl-urea a tetrafluoro borate

DIEA=N, the N-diisopropylethylamine

Use following structure I I to describe the product that obtains among the embodiment 1-7.

The detection of anti-mycotic activity and quantitative:

By using test of standard agar dilution or disk diffusion test to obtain the minimum inhibition concentration (MIC) of compound at the external test anti-mycotic activity.Used typical fungus is a Candida albicans in measuring anti-mycotic activity.When working sample (50 μ l) produces the inhibition of 10-12mm diameter region on the agar plate of Candida albicans x657 inoculation, think that anti-mycotic activity is remarkable.The tail venous toxicity:

Handled mouse with the mensuration compound (20mg/kg) of 0.1ml through vein (IV) by sidepiece tail vein at 0,24,48 and 72 hour.In each group 2 mouse are arranged.Compound is formulated in 5.0% glucose and the Injectable sterile water.After handling for the first time to mouse monitoring 7 days also close observation comprise following stimulus signal: erythema, swelling, variable color, necrosis, tail lose and any other signal of demonstration toxic side effects.

The mouse that this institute uses is that counterpoise is the male ICR mouse (can be from Harlan Sprangue Dawley, Indianapolis, IN acquisition) of the outbreed of 18-20g.Preparation preparation 4-brooethyl-5-methyl isophthalic acid, 3-dioxole-2-ketone (1a-1):

Reflux 0.1mol 4,5-dimethyl-1,3-dioxole-2-ketone, 0.1molN-bromine succinimide and 0.1g 2, the 70ml tetracol phenixin (CCl of 2-azo two (2-methyl propionitrile) ₄) mixture.After 6 hours, this mixture is with ice-cooled and filter.Filtrate is with 2 * 50ml water, 2 * 50ml sodium chloride solution and another 50ml water washing.Solution with dried over sodium sulfate and be evaporated to dried, the dry oil that obtains 16.5g (85% productive rate) under vacuum, it has the 1H-NMR data consistent with structure 1a-1.

Preparation compound (1a-2):

Compound 1a-2 is to use Synthetic Communication, and 22 (9), the method synthetic described in 1297 (1992), the crude product oil of acquisition 11.5g (78% productive rate).

Preparation compound 1a-3:

Under 4 ℃, 11.5g compound 1a-2 (crude product oil), 500ml 37%HCl and the stirring of 300ml methanol mixture are spent the night.Then this mixture is concentrated and form oil.Obtain 5.27g (33.8%) product by column chromatography (1: 1 ethyl acetate/hexane) purification, it has the 1H-NMR data consistent with structure 1a-3.

Preparation compound 1a-4:

The 30ml chloroform mixture of 3.0g compound 1a-3 and 2.02g pyridine is cooled to 0-4 ℃.Join the 30ml chloroformic solution of 5.08g p-nitrophenyl chloroformate ester in this mixture and stir about 4.5 hours.This mixture with cold 1% sodium hydroxide (3 * 30ml), 1NHCl (2 * 30ml), water (2 * 30ml) and salt solution (2 * 30ml) washing.This solution with dried over sodium sulfate, filter and use washed with dichloromethane.Removing desolvates obtains leaving standstill solidified oil.This solid is put into the 10ml methylene dichloride and add hexane formation precipitation.Mixture is filtered, use hexane wash, and under vacuum dried overnight, the product of acquisition 6.39g (94% productive rate), its 1H-NMR data are consistent with the structure of 1a-4.

Preparation compound 1b-1: Compound 1b-1 can use Synthesis, and the method described in 1159 (1990) is synthetic.Preparation compound b-2:

40ml dichloromethane solution with 4.57g (30mmol) crude product 1b-1 under 0 ℃ joins in the 100ml dichloromethane solution of 3.91g (34mmol) N-hydroxy-succinamide and 2.7g (34mmol) pyridine.After stirring 30 minutes under 0 ℃, at room temperature with this mixture standing over night.Mixture washes with water four times and then with the organic phase dried over sodium sulfate.After the filtration, solvent evaporated, the oily crude product of acquisition 4.0g (58% productive rate), its 1H-NMR data are consistent with the structure of 1b-2.The pseudobactin B (2a-1) of preparation CBZ-protection:

Pseudobactin B is dissolved/is suspended among the DMF (20mg/ml, Aldrich Sure Seal).Add N-(benzyloxycarbonyloxy base) succinimide (6eq) when at room temperature stirring.Stirred 32 hours under the room temperature.By HPLC (4.6 * 50mm, 3.5 μ m, 300-SB, C8, Zorbax post) monitoring reaction.At room temperature on the high vacuum rotatory evaporator, reactant is concentrated into 10ml.Material is put into refrigerator up to being ready to preparative chromatography.After preparing HPLC purifying and lyophilize, anti-phase obtains an amorphous white solid (compound 2a-1).Preparation compound 2b-1:

R ^{1 '}, R ^{1 "}And R ¹ =H

R ²=-NH (cyclopropyl)

R ³＝-OH

2b-1

(400mg 0.25mmo1) is dissolved in the 4ml dry DMF pseudobactin B (2a-1) that CBZ-is protected.Add successively TBTU (79mg, 0.25mmol), DIEA (200 μ l, 6 equivalents) and cyclopropylamine (14.2mg, 0.25mmol).Stirring reaction under room temperature, nitrogen is monitored by HPLC simultaneously.Under vacuum, reactant is concentrated after finishing.Crude product is purified by preparation HPLC.Lyophilize obtains 209.2mg (51.1%) colourless powder.

Under the hydrogen capsule, use the described 3-amido compounds of 10%Pd/C catalytic hydrogenation in 1%HOAc/MeOH (279.1mg, 0.169mmo1) 45 minutes.Reactant is filtered and under vacuum, concentrate.With resistates with 1: 1 water: the ACN mixture process, lyophilize obtains 208.3mg (98.6%) colourless powder (2b-1) then.This structure is passed through H ¹-NMR confirms.Preparation compound 3a-1:

R ^{1 '}, R ^{1 "}And R ¹ =H

R ²＝-OCH ₃

R ³＝-OCH ₃

3a-1

The pseudobactin B (2a-1) of adding 10ml absolute ethanol and 251.7mg CBZ-protection (0.156mmol) in the 50ml round-bottomed flask.Adding about 1ml acidifying ethanol (using HCl gas to shift to an earlier date acidifying) in this mixture also at room temperature spends the night the reactant stirring.Undertaken next step by it being dissolved in the 10ml MeOH/1.5ml ice AcOH solution under the situation of under vacuum, solvent being removed then and resistates not being had further to purify.Used 249.7mg 10%Pd/C standard hydrogenolysis 30 minutes, and removed by filter catalyzer and, obtain the compound 3a-1 of 120.9mg after the lyophilize through preparation HPLC purification.C ₅₅H ₉₆ClN ₁₂O ₁₉(M+H) ⁺MS (Ionspray) calculated value be 1264.89, experimental value is 1264.3.Preparation C-18 side chain (4a-1):

((10.9mL 68.69mmol), then adds pure TiCl to add trimethylammonium allyl group silicomethane among the 6.22g, dichloromethane solution 19.1mmol) (190mL) to chirality acetal 4a-1 under-78 ℃ ₄(2.94mL, 26.71mmol).Reactant stirred 1 hour down at-78 ℃, stirred 2 hours down at-40 ℃ then.At this moment, reaction stops with methyl alcohol (15mL) and dilutes with methylene dichloride (200mL).The gained reaction mixture with 1N HCl (2 * 50mL), water and salt water washing.Organic layer concentrates through super-dry and under vacuum and obtains a resistates, and it is purified by silica gel chromatography (10%EtOAc/ hexane), obtains the purpose product 4b-1 of 5.51g (78%).

To 4b-1 (8.56g, add in methylene dichloride 23.3mmol) (155mL) solution PCC (10.0g, 46.5mmol).Reactant at room temperature stirred 18 hours, filtered by a Celite pad then.Filtrate concentrates under vacuum and obtains a blush resistates, and it is purified by silica gel chromatography (10%EtOAc/ hexane), obtains 8.36g (80%) methyl ketone intermediate (structure does not show).(8.36g 22.8mmol) is dissolved among THF (60mL) and the MeOH (30mL) with the intermediate that obtains here.In this solution, add 7.5M KOH (15mL).After stirring 3 hours under the room temperature, remove partial solvent.Residue reaction mixture EtOAc/Et ₂O (3: 1 ratios, 350mL) dilution.Organic layer water (3 * 50mL) and the salt water washing.The gained organic layer concentrates through super-dry and under vacuum and obtains resistates, and it is purified by silica gel chromatography (10%EtOAc/ hexane), obtains the purpose white solid product 4c-1 of 6.22g (96%).

(6.22g 22.0mmol) is dissolved in the THF aqueous solution (5.5mL water and 55mL THF) with methyl alcohol 4c-1.(4.42g 33.0mmol), then adds OsO to add NMO in this solution ₄(280mg is dissolved in THF, 1.10mmol).At room temperature stirring reaction spends the night.Add sodium disulfide (4g) this moment.Stirring reaction 2 hours is used EtOAc (300mL) dilution then.Whole mixture water (2 * 40mL) and the salt water washing.The gained organic layer concentrates through super-dry and under vacuum and obtains corresponding three alcohol intermediates.This material is dissolved in MeOH (200mL) and the water (40mL).In this solution, add NaIO ₄(10.6g, 49.5mmol).After stirring 1 hour under the room temperature, reactant filters by Celite and purifies by short column silica gel chromatography (30%EtOAc/ hexane), obtains about 10g (＞100%) crude product beta-hydroxy aldehyde.Thus obtained impure aldehyde is dissolved in the trimethyl carbinol (100mL) and the tetrahydrobenzene (14mL).In this solution, add NaClO under the room temperature ₂(15.97g, 176mmol) and KH ₂PO ₄(17.8g, aqueous solution 132mmol) (50mL).Reactant at room temperature stirred 6 hours, used 5N HCl termination reaction to pH=4 down at 0 ℃ then.Reactant was with 3: 1 EtOAc/Et ₂O (3 * 250mL) mixed extractant solvents.Organic layer obtains 7.3g (＞100%) crude acid 4d-1 with salt water washing and dry and concentrated, and it is directly used in coupled reaction.Preparation pseudobactin C-18 (compound 4b-2):

4a-2 R＝Cbz

4b-2 R＝H

(2.1g 6.99mmol) is dissolved among anhydrous THF (20mL) and the DMF (20mL) with crude acid 4d-1.In this solution, add HOBt (1.23g, 9.08mmol) and EDCI (1.74g, 9.08mmol).After stirring 8 hours under the room temperature, and the pseudobactin nuclear of adding CBZ-protection (3.87g, 2.80mmol).At room temperature stirring reaction is 2 days.Partly remove and desolvate this moment.With reaction mixture application of sample purify on the preparation anti-phase HPLC system (injecting 4 times).After the lyophilize, isolate the C18 acyl derivative 4a-2 and the HOBt activatory side chain ester (1g) of the CBZ-protection of 550mg (12%).(1.0g 2.40mmol) next examines (1.33g, 0.96mmol) reaction in anhydrous THF and DMF (each 10mL) to the side chain ester that reclaims with the pseudobactin of CBZ-protection.Then carry out after the above-mentioned identical purification step, obtain the C18 derivative 4a-2 (606mg, 38%) of the CBZ-protection of amount in addition.

C18 derivative 4a-2 to the CBZ-protection under-78 ℃ (adds Pd/C (550mg, 10% palladium content) among the 550mg, 10%HOAc/MeOH solution (55mL) 0.33mmol).Reactant hydrogenation 40 minutes under 1.5 normal atmosphere.By analyzing HPLC monitoring reaction process.After the end, filter out catalyzer and under vacuum, filtrate is concentrated in 30 ℃.The gained resistates is dissolved in 1: 1 once more and contains in the water-acetonitrile and lyophilize, obtains the purpose product 4b-2 of 250mg (60%).

In following each embodiment, use specific pseudobactin compound as raw material; Yet, one of skill in the art will appreciate that the pseudobactin compound that has different N-acyl group except use begins, use same steps as just can synthesize other N-acyl derivative.

Embodiment 1

Following examples illustrate pseudobactin C ' (n=12, R ²And R ³The preparation of one replacement of=-OH), two replacements or trisubstituted acyloxy alkyl carbamate prodrug.

Adding 1.5 normal compound 1a-4 in the DMF solution (1 liter) that contains pseudobactin C ' (1.5g, 1 equivalent) also at room temperature stirred the mixture about 3 days.Partly except that desolvating and resistates being passed through anti-phase HPLC (Waters ^TM, Delta Pak C18 post) and purifying obtains following product and mixture of products:

The pseudobactin C ' of the single replacement of 86mg (1-1);

The mixture (1-2) of the monobasic pseudobactin C ' of 87mg;

The mixture (1-3) of the dibasic pseudobactin C ' of 177mg;

The pure dibasic pseudobactin C ' of 132mg (1-4); With

The trisubstituted pseudobactin C ' of 248mg (1-5).With regard to trisubstituted prodrug or dibasic prodrug mixture, do not observe the stimulation of tail vein.Observe certain stimulation with single replacement, monobasic mixture and pure dibasic prodrug sample.Compare with it, unsubstituted pseudobactin C ' and unsubstituted pseudobactin B have clearly illustrated the stimulation of tail vein.(ED except pure dibasic prodrug sample ₅₀＞20mg/kgx4), all samples has all shown effect in the significant body.

A replacement, two of using above-mentioned same procedure also to prepare top structure I I replaces or trisubstituted prodrug sample, wherein R ^{1 '}, R ^{1 "}And/or R ¹ Be

And n=10,12 and 14.

Compound 1-1 presents the interior effect of the body similar to parent compound pseudobactin C ' with 1-5.Do not observe the sign of the tail venous stimulation of compound 1-5, and observe the tail venous toxicity of the raising of compound 1-1.Unexpectedly, do not observe the interior effect of body of compound 1-4.

Embodiment 2

Following examples illustrate pseudobactin B (n=10, R ²And R ³The preparation of one replacement of=-OH), two replacements or trisubstituted acyloxy alkyl carbamate prodrug.

(2.0g 1.65mmol) adds 574mg (2.47mmol) compound 1b-2 in the solution to the pseudobactin B that is dissolved in the 500ml dimethyl formamide.At room temperature stir the mixture and spend the night.Then solution concentration to about 50ml and product used following gradient elution scheme purification: 0-30%0.1%TFA/ACN 5 minutes and 30-70%TFA/ACN 40 minutes by HPLC.Total recovery 59%.

(one replaces prodrug (compound 2-1), wherein R to three separated products ^{1 '}And R ^{1 "}=H and R ¹ =-C (O) OCH ₂OAc, two replace prodrug (compound 2-2), wherein R ^{1 '}And R ^{1 "}=-C (O) OCH ₂OAc and R ¹ =H and three replaces prodrug (compound 2-3), wherein R ^{1 '}, R ^{1 "}And R ¹ =-C (O) OCH ₂OAc) all measure and confirm effect in the body of anti-mouse general mycocandida.Yet the tail venous toxicity is positive.

Embodiment 3

Use with the foregoing description 2 in identical universal method prepare pseudobactin B (n=10, R ²And R ³One of=-OH) replaces, two replacements and three replace prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) C (CH ₃) ₃Isolate following 5 samples:

3-1 one replaces R ¹

3-2 mixes monobasic R ^{1 '}And R ^{1 "}

3-3 mixes dibasic R ¹ + R ^{1 '}And R ¹ + R ^{1 "}

The dibasic R of 3-4 ^{1 '}+ R ^{1 "}

The trisubstituted R of 3-5 ^{1 '}+ R ^{1 "}+ R ¹

Sample 3-1,3-3 and 3-5 show that separately the tail venous toxicity is negative.All 5 samples all demonstrate the interior effect of body of anti-mouse general mycocandida.

Embodiment 4

Use with the foregoing description 2 in identical universal method prepare pseudobactin C ' (n=12, R ²And R ³One of=-OH) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) C (CH ₃) ₃Isolate following 5 samples:

The monobasic R of 4-1 ¹

4-2 mixes monobasic R ^{1 '}And R ^{1 "}

4-3 mixes dibasic R ¹ + R ^{1 '}And R ¹ + R ^{1 "}

The dibasic R of 4-4 ^{1 '}+ R ^{1 "}

The trisubstituted R of 4-5 ^{1 '}+ R ^{1 "}+ R ¹

There is not working sample 4-1.Sample 4-3,4-4 and 4-5 show it is tail venous toxicity feminine gender.Sample 4-2,4-3,4-4 and 4-5 demonstrate the interior effect of body of anti-mouse general mycocandida.

Embodiment 5

Use with the foregoing description 2 in identical universal method prepare that one of pseudobactin B (n=10) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH (CH ₃) OC (O) CH ₃Only measured trisubstituted derivative (compound 5-1).Described trisubstituted compound exhibits tail venous toxicity is negative.

Embodiment 6

Use with the foregoing description 2 in identical universal method prepare pseudobactin B (n=10, R ²And R ³One of=-OH) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) CH ₂CH ₃Only measured trisubstituted derivative (compound 6-1).Described trisubstituted compound exhibits effect in the body of good anti-mouse general mycocandida, and do not have the tail venous stimulation.

Embodiment 7

Use with the foregoing description 2 in identical universal method prepare pseudobactin B (n=10, R ²And R ³One of=-OH) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) CH (CH ₃) CH ₃Only measured trisubstituted derivative (compound 7-1).Described trisubstituted compound exhibits effect in the body of good anti-mouse general mycocandida, and do not have the tail venous stimulation.

Embodiment 8

Embodiment 8 and 9 has described by the synthetic prodrug of semisynthetic pseudobactin compound, wherein the unitary N-acyl group of the L-Serine in the pseudobactin structure is changed.

Use with the foregoing description 2 in identical universal method prepare pseudobactin C-18 (n=14, R ²And R ³One of=-OH) (4b-2) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) C (CH ₃) ₃Only measured trisubstituted derivative (compound 8-1).Described trisubstituted compound exhibits tail venous toxicity is negative.

Embodiment 9

Use with the foregoing description 2 in identical universal method prepare pseudobactin C-18 (n=14, R ²And R ³One of=-OH) (4b-2) replaces, two replacement and trisubstituted prodrug, wherein R ^{1 '}, R ^{1 "}And/or R ¹ =-C (O) OCH ₂OC (O) CH (CH ₃) CH ₃Only measured trisubstituted derivative (compound 9-1).Described trisubstituted compound exhibits negative tail venous toxicity.

Embodiment 10

Embodiment 10 has described the further modification of aforementioned prodrugs, wherein with the single acylamino derivative of modified formation one 3-of the hydroxy-acid group of the aspartic acid units of pseudomonas prime ring.Compound 10-1,10-2,10-3,10-4 and 10-5's is synthetic:

R ^1′、R ^1″、R ¹＝-C(O)OCH ₂OC(O)C(CH ₃) ₃

R ²＝-NHCH ₂CH ₂N(CH ₃) ₂

R ³＝-OH

10-1

To prodrug 3-5 (864mg, add in the DMF solution (9ml) 0.52mmol) 1-dimethylamino-2-ethylamine (57.9 μ l, 0.52mmol) and TBTU (168.6mg 0.52mmol), then adds diisopropylethylamine (423 μ l).After stirring 20 minutes under the room temperature, reaction mixture is purified by reversed-phase HPLC (ACN:0.1%TFA/ water).Lyophilize obtains the compound 10-1 of 295mg (34%).

R ^1′、R ^1″、R ¹＝-C(O)OCH ₂OC(O)C(CH ₃) ₃

R ²=-NH (cyclopropyl)

R ³＝-OH

10-2

Compound 10-2 is to use and top same procedure synthetic, but uses the 0.052mmol cyclopropylamine to replace 1-dimethylamino-2-ethylamine.

R ^1′、R ^1″、R ¹＝-C(O)OCH ₂OC(O)C(CH ₃) ₃

R ²＝-NHCH ₂(CO ₂CH ₃)

R ³＝OH

10-3

Compound 10-3 is to use and top same procedure synthetic, but uses glycine methyl ester to replace 1-dimethylamino-2-ethylamine.

R ^1′、R ^1″、R ¹＝-C(O)OCH ₂OC(O)CH(CH ₃) ₂

R ²＝-NHCH ₂CH ₂N(CH ₃) ₂

R ³＝-OH

10-4

Compound 10-4 is to use and top same procedure synthetic, but uses 0.052mmol prodrug 7-1 to replace prodrug 3-5.

R ^1′、R ^1″、R ¹＝-C(O)OCH ₂OC(O)CH(CH ₃) ₂

R ²=-NH (cyclopropyl)

R ³＝-OH

10-5

Compound 10-5 is to use and top same procedure synthetic, but uses 0.052mmol prodrug 7-1 replacement prodrug 3-5 and use the 0.052mmol cyclopropylamine to replace 1-dimethylamino-2-ethylamine.

Embodiment 11

Embodiment 11 has described the synthetic of pseudobactin compound prodrug, generates the 3-acylamino derivative thereby wherein the hydroxy-acid group of the aspartic acid units of pseudomonas prime ring is modified.The single acylamino derivative 11-1 of synthetic 3-:

R ^{1 '}, R ^{1 "}With

R ²=-NH (cyclopropyl)

R ³＝-OH

11-1

Adding 1.5 normal compound 1a-4 in the DMF solution that contains compound 2b-1 (1 equivalent) (1 liter) also at room temperature stirred the mixture about 3 days.Partly except that desolvating and resistates being passed through reversed-phase HPLC (Waters ^TM, Delta Pak C18 post) purify, obtain compound 11-1 and other replacement and a dibasic product.

Embodiment 12

Embodiment 12 has described the synthetic of prodrug, wherein the hydroxy-acid group on aspartic acid and the hydroxyl aspartic acid is modified the generation diester derivatives.Dibasic acid esters 12-1's is synthetic:

R ^{1 '}, R ^{1 "}With

R ²＝-OCH ₃

R ³＝-OCH ₃

12-1

Adding 1.5 normal compound 1a-4 in the DMF solution that contains compound 3a-1 (1 equivalent) (1 liter) also at room temperature stirred the mixture about 3 days.Partly except that desolvating and resistates being passed through reversed-phase HPLC (Waters ^TM, Delta Pak C18 post) purify, obtain compound 12-1 and other replacement and a dibasic product.

Claims (16)

1. Pseudomycin prodrugs, its pharmacologically acceptable salt and solvate thereof with following structure,

Wherein R is

Wherein

R ^aAnd R ^{A '}Be H 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 H, halogen or methyl, perhaps R independently ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl;

R ^cBe H, 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 H, perhaps with R ^fBe 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, C ₅-C ₁₁Alkoxyl group or xenyl; R is

Wherein

R ^gBe H 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 H, 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 H ,-CH ₃Or-C (O) CH ₃

R ¹Be H, acyloxy methylene radical-1 independently, 3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters, condition is at least one R ¹Be acyloxy methylene radical-1,3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters;

R ²And R ³Be independently-OR ^2a, or-N (R ^2b) (R ^2c),

Wherein

R ^2aAnd R ^2bBe H, C independently ₁-C ₁₀Alkyl, C ₃-C ₆Cycloalkyl, hydroxyl (C ₁-C ₁₀) alkyl, alkoxyl group (C ₁-C ₁₀) alkyl, C ₂-C ₁₀Alkenyl, amino (C ₁-C ₁₀) alkyl ,-or two-alkylamino (C ₁-C ₁₀) alkyl, aryl (C ₁-C ₁₀) alkyl, heteroaryl (C ₁-C ₁₀) alkyl, the assorted alkyl (C of ring ₁-C ₁₀) alkyl, perhaps

R ^2bBe the alkyl carboxylates residue and the R of aminoacid alkyl ester ^2cBe H or C ₁-C ₆Alkyl.

2. the prodrug of claim 1, wherein said acyloxy methylene radical-1,3-dioxole-2-ketone is represented by following formula 1 (a):

R wherein ^1aBe C ₁-C ₁₀Alkyl, C ₁-C ₁₀Thiazolinyl, benzyl or aryl, and R ^1bBe H or methyl.

3. the prodrug of claim 1, wherein said acyloxy methylene radical carboxylicesters is represented by following formula 1 (b):

R wherein ^1aBe C ₁-C ₁₀Alkyl, C ₁-C ₁₀Thiazolinyl, benzyl or aryl, and R ^1bBe H or methyl.

4. the prodrug of claim 2, wherein R is represented by following structure:

R wherein ^{B '}Be hydroxyl, R ^a, R ^{A '}, R ^b, R ^c, R ^dAnd R ^eAll be H, and R ^fIt is n-octyl.

5. the prodrug of claim 3, wherein R is represented by following structure: R wherein ^{B '}Be hydroxyl, R ^a, R ^{A '}, R ^b, R ^c, R ^dAnd R ^eAll be H, and R ^fIt is n-octyl.

6. the prodrug of claim 1, the alkyl carboxylates residue of wherein said aminoacid alkyl ester is by following expression :-CH ₂CO ₂CH ₃,-CH (CO ₂CH ₃) CH (CH ₃) ₂,-CH (CO ₂CH ₃) CH (phenyl) ,-CH (CO ₂CH ₃) CH ₂OH ,-CH (CO ₂CH ₃) CH ₂(p-hydroxybenzene) ,-CH (CO ₂CH ₃) CH ₂SH ,-CH (CO ₂CH ₃) CH ₂(CH ₂) ₃NH ₂,-CH (CO ₂CH ₃) CH ₂(4-imidazoles) ,-CH (CO ₂CH ₃) CH ₂(5-imidazoles) ,-CH (CO ₂CH ₃) CH ₂CO ₂CH ₃, or-CH (CO ₂CH ₃) CH ₂CO ₂NH ₂

7. Pseudomycin prodrugs, its pharmacologically acceptable salt and solvate thereof with following structure, Wherein R is Wherein

R ^aAnd R ^{A '}Be H 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 H, halogen or methyl, perhaps R independently ^bOr R ^{B '}Be amino, alkylamino, α-acetylacetic ester, methoxyl group or hydroxyl;

R ^cBe H, 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 H, perhaps with R ^fBe 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, C ₅-C ₁₁Alkoxyl group or xenyl; R is Wherein

R ^gBe H 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 H, 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 H ,-CH ₃Or-C (O) CH ₃

R ¹Be H, acyloxy methylene radical-1 independently, 3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters, condition is at least one R ¹Be acyloxy methylene radical-1,3-dioxole-2-ketone or acyloxy methylene radical carboxylicesters;

R ²And R ³Be independently-OR ^2a, or-N (R ^2b) (R ^2c),

Wherein

R ^2aAnd R ^2bBe H, C independently ₁-C ₁₀Alkyl, C ₃-C ₆Cycloalkyl, hydroxyl (C ₁-C ₁₀) alkyl, alkoxyl group (C ₁-C ₁₀) alkyl, C ₂-C ₁₀Thiazolinyl, amino (C ₁-C ₁₀) alkyl ,-or two-alkylamino (C ₁-C ₁₀) alkyl, aryl (C ₁-C ₁₀) alkyl, heteroaryl (C ₁-C ₁₀) alkyl, the assorted alkyl (C of ring ₁-C ₁₀) alkyl, perhaps

R ^2bBe the alkyl carboxylates residue and the R of aminoacid alkyl ester ^2cBe H or C ₁-C ₆Alkyl.

8. the prodrug of claim 7, wherein said acyloxy methylene radical-1,3-dioxole-2-ketone is represented by following formula 1 (a):

R wherein ^1aBe C ₁-C ₁₀Alkyl, C ₁-C ₁₀Thiazolinyl, benzyl or aryl, and R ^1bBe H or methyl.

9. the prodrug of claim 7, wherein said acyloxy methylene radical carboxylicesters is represented by following formula 1 (b):

R wherein ^1aBe C ₁-C ₁₀Alkyl, C ₁-C ₁₀Thiazolinyl, benzyl or aryl, and R ^1bBe H or methyl.

10. the prodrug of claim 8, wherein R is represented by following structure: R wherein ^{B '}Be hydroxyl, R ^a, R ^{A '}, R ^b, R ^c, R ^dAnd R ^eAll be H, and R ^fIt is n-octyl.

11. the prodrug of claim 9, wherein R is represented by following structure: R wherein ^{B '}Be hydroxyl, R ^a, R ^{A '}, R ^b, R ^c, R ^dAnd R ^eAll be H, and R ^fIt is n-octyl.

12. the prodrug of claim 7, the alkyl carboxylates residue of wherein said aminoacid alkyl ester is by following expression :-CH ₂CO ₂CH ₃,-CH (CO ₂CH ₃) CH (CH ₃) ₂,-CH (CO ₂CH ₃) CH (phenyl) ,-CH (CO ₂CH ₃) CH ₂OH ,-CH (CO ₂CH ₃) CH ₂(p-hydroxybenzene) ,-CH (CO ₂CH ₃) CH ₂SH ,-CH (CO ₂CH ₃) CH ₂(CH ₂) ₃NH ₂,-CH (CO ₂CH ₃) CH ₂(4-imidazoles) ,-CH (CO ₂CH ₃) CH ₂(5-imidazoles) ,-CH (CO ₂CH ₃) CH ₂CO ₂CH ₃, or-CH (CO ₂CH ₃) CH ₂CO ₂NH ₂

13. each compound of aforementioned claim is used for resisting the purposes of the medicine of systemic fungal infection or fungal skin infections in preparation.

14. a medicinal preparations contains Pseudomycin prodrugs and pharmaceutically acceptable carrier of claim 1 or 7.

15. a medicine for the treatment of the animal anti-fungal infection, wherein said medicine contains the compound of claim 1 or 7.

16. a method for the treatment of the animal anti-fungal infection that needs treatment comprises to described animals administer claim 7,8,9,10 or 11 Pseudomycin prodrugs.