Desleucyl glycopeptide antibiotics and methods of making same

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] The present invention relates to glycopeptide compounds having antibiotic activity, and methods of making glycopeptide compounds having antibiotic activity.

[0003] 2. Background of the Invention

[0004] Glycopeptide antibiotics are characterized by having at least one saccharide group chemically bonded to a rigid peptide structure having a cavity or cleft which acts as a binding site for the substrate used in bacterial cell wall synthesis. The glycopeptide antibiotics are further categorized into various subclasses depending on the identity and interconnections of the amino acids comprising the peptide backbone and the number and substitution pattern of the sugar residues in the molecule. The glycopeptide antibiotics are generally active against Gram-positive bacteria but relatively ineffective against Gram-negative bacteria. Most notable among the glycopeptide antibiotics is vancomycin. Vancomycin is produced by Amycolatopsis orientalis, and is often referred to as “the drug of last resort” because it is effective against most multi-drug-resistant gram positive bacteria. However, in recent years, vancomycin-resistant strains of some bacteria have emerged.

[0005] The structural formula of vancomycin is shown below and is characterized by a disaccharide moiety covalently linked to a heptapeptide structure. The structure of vancomycin places it in a class of molecules referred to as the “dalbaheptides.” [Malabarba A., et al. (1997a)]. Dalbaheptides in general are characterized by the presence of seven amino acids linked together by peptide bonds and held in a rigid conformation by cross-links through the aromatic substituent groups of at least five of the amino acid residues. In the heptapeptide structure of vancomycin, which is commonly referred to as the “aglycone” of vancomycin, the aromatic side-chains of amino acids 2, 4, and 6 are fused together through ether linkages. The aromatic side-chains of amino acids 5 and 7 are joined via a carbon-carbon bond. Amino acids 1 and 3 are N-methyl leucine and asparagine, respectively. Other naturally-occurring glycopeptide antibiotics are similar to vancomycin in that they have the same amino acids 1 through 7 forming the peptide binding pocket and a glucose residue linked to the aromatic substituent on amino acid 4 through formation of a bond with a phenolic hydroxyl group. The glucose residue, in turn, is linked through its vicinal hydroxyl position to a unique amino sugar, L-vancosamine. Some glycopeptide antibiotics similar to vancomycin contain additional glycosidic groups attached to other positions on the peptide (e.g. chloroeremomycin). Still other glycopeptide antibiotics such as β-avoparcin are similar to vancomycin in that they contain the same amino acids at all positions except positions one and three. β-avoparcin, for example, contains an amino acid containing an aromatic side chain in place of the asparagine at position three and does not contain N-methyl leucine at position one. β-avoparcin contains glycosidic groups at amino acid 4 and at other positions on the peptide core.

[0006] Vancomycin, chloroeremomycin and β-avoparcin have the structures as shown below:

[0007] Eremomycin has the structure of chloroeremomycin except that the chlorine substituent on the aromatic group attached to amino acid 6 is not present in eremomycin.

[0008] The anti-microbial activity of the naturally occurring glycopeptide antibiotics is believed to be due to their ability to interfere with biosynthesis of the bacterial cell wall, evidently by binding to dipeptide termini of uncross-linked peptidoglycan and/or the disaccharide precursor of peptidoglycan. [Nagarajan R. (1993)]. NMR evidence has shown that the heptapeptide chain of vancomycin forms a number of hydrogen bonds with D-alanyl-D-alanine, the dipeptide that is at the terminus of the peptide chain attached to the N-acetylmuramic acid unit that is incorporated into peptidoglycan. [See, e.g., Prowse W., et al. (1995); Pierce C., et al. (1995); Williams D. et al. (1998)]. The interaction of vancomycin with peptidoglycan precursors apparently inhibits or prevents the subsequent transglycosylation and/or transpeptidation steps of cell wall assembly. Supporting this mode of action is the fact that vancomycin-resistant strains of bacteria are found to produce a pentapeptide precursor terminating in a D-alanyl-D-lactate sequence. It is hypothesized that the reduced effectiveness of vancomycin against resistant strains is due to reduced hydrogen bonding interactions between the drug and the D-alanyl-D-lactate substrate (and possibly repulsive interactions as well). The affinity of vancomycin for D-alanyl-D-lactate is estimated to be 2-3 orders of magnitude (4.1 kcal/mol) less than for D-alanyl-D-alanine. [Walsh C (1993)].

[0009] The sugar residues of vancomycin and other glycopeptide antibiotics have been shown to affect biological activities. Structural changes in the sugar residues can produce significant changes in antibiotic activity. [Malabarba (1997); Nagarajan, R. (1993)]. It has been proposed that the sugar residues on the glycopeptide antibiotics may enhance the avidity of these molecules for surface-bound peptide ligands. At least two different mechanisms for enhancing avidity have been proposed. [Kannan (1998); Gerhard (1993); Allen (1997)].

[0010] For example, it has been proposed that the biological activity of vancomycin, along with that of many other glycopeptide antibiotics, is enhanced by dimerization [Williams D., et al. (1993); Gerhard, U., et al., (1993)] facilitated by the saccharide groups on the convex surface of the molecules. Structural evidence for dimerization of several different glycopeptides has been obtained from both NMR and crystallographic studies. It has been found that there are significant differences in the stability of the dimers formed in solution by different glycopeptide antibiotics. [MacKay (1994)]. Dimerization is thought to influence activity by increasing the avidity of the glycopeptides for surface-bound D-ala-D-ala peptide ligands [Williams, (1998)]. It is proposed that the differences in the dimerization constants, due to different interactions between saccharide groups, may account at least partially for the differences in biological activity of different glycopeptide antibiotics which otherwise have very similar peptide binding pockets and also have similar affinities for the natural D-ala-D-ala substrate. [Williams (1998)].

[0011] A second mechanism for enhancing activity has been proposed for the naturally occurring glycopeptide antibiotic teicoplanin and various semi-synthetic glycopeptides containing hydrophobic substituents on at least one of the saccharide units. It is suggested that hydrophobic substituents (a C2 N-acyl group in the case of teicoplanin) interact with the bacterial membrane, thus “anchoring” hydrophobically substituted glycopeptides at the membrane surface. [Beauregard (1995)]. Membrane anchoring is proposed to enhance activity by localizing the glycopeptide antibiotic to the membrane where the Lipid II substrates that are the precursors of peptidoglycan are found. The glycopeptide antibiotics then bind to the dipeptide termini of these precursors and prevent transglycosylation and/or transpeptidation.

[0012] It should be noted that teicoplanin is active against some vancomycin resistant strains. Furthermore, the attachment of hydrophobic substituents to the vancomycin carbohydrate moiety confers activity against these and other vancomycin-resistant bacterial strains. [Nagarajan (1991)]. It has been speculated that the lipophilic groups on the saccharides, in locating the antibiotic at the cell surface, help overcome the decreased binding affinity for D-ala-D-lac in vancomycin resistant microorganisms.

[0013] It has generally been assumed that peptide binding is essential for biological activity. In fact, it had been shown that if the peptide core of vancomycin is damaged by removing the N-methyl leucine (amino acid 1), the resulting compound loses affinity for D-ala-D-ala and has no biological activity, even against sensitive bacterial strains. The lack of biological activity is presumed to be due to the inability of the compound to bind D-Ala-D-Ala well.

[0014] Previously, others have explored the possibility of attaching amino acids other than N-methyl leucine to the amino acid 2 on des-N-methyl leucyl vancomycin. It was found that some amino acid substitutions produced compounds with comparable activity to vancomycin; some had worse activity. No useful improvements in activity have been reported. As far as we know, no substitutions have ever been made at the A1 position of any dalbaheptides wherein A1 and A3 are not directly linked by a covalent bond and wherein there is at least one hydrophobic substituent on at least one of the sugar moieties attached to at least one of the amino acids A2-A7 Thus, replacing N-methyl leucine at A1 on vancomycin with other amino acids did not yield any compounds having significantly better properties than vancomycin itself. Hence, it would not be expected that glycopeptides having at least one hydrophobic substituent attached to a glycosidic group on any one of amino acids A2-A7 and having no A1 group or an A1 group other than an N-substituted leucine would have any antibiotic properties, much less better antibiotic properties than the precursor glycopeptide compounds.

[0015] Definitions

[0016] A “glycoconjugate” comprises any molecule linked to at least one carbohydrate of any size. The molecule can be a peptide or protein, a nucleic acid, a small molecule, a lipid, or another carbohydrate; it may be of natural or non-natural origin.

[0017] A “glycopeptide” is a glycoconjugate comprising a peptide linked to at least one carbohydrate.

[0018] A “glycopeptide antibiotic” is a glycopeptide having antibacterial activity, including, e.g., vancomycin, eremomycin, chloroeremomycin and β-avoparcin as well as any synthetic and semi-synthetic derivatives thereof. The term “glycopeptide antibiotic” is meant to encompass any naturally occurring antibiotic as well semi-synthetic derivatives thereof.

[0019] An “aglycone” is the result of removing the carbohydrate residues from a glycopeptide, leaving only a peptide core.

[0020] A “des-N-methyl leucyl aglycone” is the result of removing a terminal N-methyl leucine residue from an aglycone.

[0021] A “pseudoaglycone” is the result of removing only one of two sugar residues from a disaccharide residue linked to amino acid residue A4 of a glycopeptide. Thus, a pseudoaglycone comprises an aglycone in which A4 is linked to a monosaccharide residue.

[0022] A “des-N-methyl leucyl pseudoaglycone” is the result of removing a terminal N-methyl leucine residue from an pseudoaglycone. Thus, a des-N-methyl-leucyl pseudoaglycone is an aglycone in which A4 is linked to a monosaccharide residue and which has a terminal N-methyl leucine residue removed therefrom.

[0023] A “dalbaheptide” is a glycopeptide containing a heptapeptide moiety which is held in a rigid conformation by cross-links between the aromatic substituent groups of at least five of the seven α-amino acid residues, including a cross-link comprising a direct carbon-carbon bond between the aryl substituents of amino acid residues 5 and 7, and aryl ether cross-links between the substituents of amino acid residues 2 and 4, and 4 and 6. Amino acid residues 2 and 4-7 in different dalbaheptides are those found in the naturally occurring glycopeptide antibiotics. These amino acid residues differ only in that residues 2 and 6 do not always have a chlorine substituent on their aromatic rings, and in that substitution on free hydroxyl or amino groups may be present. Amino acids residues 1 and 3 may differ substantially in different dalbaheptides; if both bear aryl substituents, these may be cross-linked. Molecules having a dalbaheptide structure include, e.g., the glycopeptide antibiotics mentioned above.

[0024] The term “alkyl” refers to a linear or branched acyclic or non-aromatic cyclic group having form one to twenty carbon atoms connected by single or multiple bonds. Thus, the term “alkyl” is meant to encompass linear or branched acyclic or non-aromatic groups having one or more carbon-carbon double or triple bonds, i.e., alkenyl and alkynyl groups. An alkyl group may be substituted by one or more of halo, hydroxyl, protected hydroxyl, amino, protected amino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, COOH, aroyloxy, alkylamino, dialkylamino, trialkylammonium, alkylthio, arylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, heterocyclic, CONH2, CONH-alkyl, CON(alkyl)2, COO-aralkyl, COO-aryl or COO-alkyl.

[0025] The term “aryl” refers to a group derived from a non-heterocyclic aromatic compound having from six to twenty carbon atoms and from one to four rings which may be fused or connected by single bonds. An aryl group may be substituted by one or more of alkyl, aralkyl, heterocyclic, heterocyclic-alkyl, heterocyclic-carbonyl, halo, hydroxyl, protected hydroxyl, amino, protected amino, hydrazino, alkylhydrazino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, aroyloxy, alkylamino, dialkylamino, trialkylammonium, alkylthio, arylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, COO-alkyl, COO-aralkyl, COO-aryl, CONH2, CONH-alkyl or CON(alkyl)2.

[0026] The term “aralkyl” refers to an alkyl group substituted by an aryl group. Aralkyl may optionally be substituted with one or more of the groups with which alkyl or aryl may be substituted.

[0027] The term “heterocyclic” refers to a group derived from a heterocyclic compound having from one to four rings, which may be fused or connected by single bonds; said compound having from three to twenty ring atoms which may be carbon, nitrogen, oxygen, sulfur or phosphorus. A heterocyclic group may be substituted by one or more alkyl, aryl, aralkyl, halo, hydroxyl, protected hydroxyl, amino, hydrazino, alkylhydrazino, arylhydrazino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, aroyloxy, alkylamino, dialkylamino, trialkylamino, alkylthio, arylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, COO-alkyl, COO-aralkyl, COO-aryl, CONH2, CONH-alkyl or CON(alkyl)2.

[0028] The terms “alkoxy,” “aryloxy,” and “aralkyloxy,” refer to groups derived from bonding an oxygen atom to an alkyl, aryl or aralkyl group, respectively. Any alkoxy, aryloxy or aralkyloxy group may optionally be substituted with one or more of the groups with which alkyl, aryl or aralkyl may be substituted. The terms “alkanoyl,” “aroyl,” and “aralkanoyl” refer to groups derived from bonding a carbonyl to an alkyl, aryl or aralkyl group, respectively. Any alkanoyl, aroyl or aralkanoyl group may optionally be substituted with one or more of the groups with which alkyl, aryl or aralkyl may be substituted. The terms “heterocyclic-alkyl” and “heterocyclic-carbonyl” refer to groups derived from bonding a heterocyclic group to an alkyl or a carbonyl group, respectively. An heterocyclic-alkyl or heterocyclic-carbonyl group may optionally be substituted with one or more of the groups with which heterocyclic or alkyl may be substituted. The term “heterocyclic-alkyl-carbonyl” refers to a group derived from bonding a heterocyclic-alkyl group to a carbonyl group. Any heterocyclic-alkyl-carbonyl may optionally be substituted with one or more of the groups with which heterocyclic or alkyl may be substituted. The term “alkylsulfonyl” refers to a group derived from bonding an alkyl group to a sulfonyl group. An alkylsulfonyl group may optionally be substituted with one or more groups with which alkyl may be substituted. The term “arylsulfonyl” refers to a group derived from bonding an aryl group to a sulfonyl group. An arylsulfonyl group may optionally be substituted with one or more groups with which aryl may be substituted. The term “protected hydroxyl” refers to a hydroxyl group bonded to a group which is easily removed to generate the free hydroxyl group by treatment with acid or base, by reduction or by exposure to light, or by any other conventional means for removing a protecting group from a hydroxyl group. The term “protected amino” refers to an amino group bonded to a group which is easily removed to generate the free amino group by treatment with acid or base, by reduction or exposure to light, or by any other conventional means for removing a protecting group from an amino group.

[0029] A “chemical library” is a synthesized set of compounds having different structures. The chemical library may be screened for biological activity to identify individual active compounds of interest.

[0030] The term “DMF” refers to N,N-dimethylformamide; “THF” refers to tetrahydrofuran; “TFA” refers to trifluoroacetic acid; “EtOAc” refers to ethyl acetate; “MeOH” refers to methanol, “MeCN” refers to acetonitrile; “Tf” refers to the trifluoroacetyl group; “DMSO” refers to dimethyl sulfoxide; “DIEA” refers to diisopropylethylamine; “All” in structural formulas refers to the allyl group; “Fmoc” refers to 9-fluorenylmethyloxycarbonyl; “HOBt” refers to 1-hydroxybenzotriazole and “Obt” to the 1-oxybenzotriazolyl group; “PyBOP” refers to benzotriazol-1-yl-oxyatripyrrolidine-phosphonium hexafluorophosphate; “Su” refers to the succinimidyl group; “HBTU” refers to O-benzoatriazol-1-yl-N2N3N′,N′-tetramethyluronium hexafluorophosphate; “alloc” refers to allyloxycarbonyl; and “CBZ” refers to benzyloxycarbonyloxy.

[0031] The term “hydrophobic” as used herein to describe a compound of the present invention or a substituent thereon, refers to the tendency of the compound or substituent thereon to lack an affinity for, to repel or to fail to absorb water, or to be immiscible in water. The term “hydrophobic” is not mean to exclude compounds or substituents thereon that are not completely immiscible in water.

[0032] The term “polar” as used herein to describe a compound of the present invention or a substituent thereon, refers to the tendency of the compound or substituent thereon to have an affinity for, to attract or to absorb water, or to be miscible in water. The term “polar” is not meant to exclude compounds or substituents thereon that are not completely miscible in water.

SUMMARY OF THE INVENTION

[0033] Thus, the present invention is directed to compounds of the formula A1-A2-A3-A4-A5-A6-A7 wherein each of the groups A2 to A7 comprises a modified or unmodified α-amino acid residue, A1 is optional and, when present, comprises an organic group other than N-substituted leucine, and at least one of the groups A2 to A7 is linked via a glycosidic bond to one or more glycosidic groups each having one or more sugar residues, wherein at least one of said sugar residues is modified to bear at least one hydrophobic substituent. In preferred compounds of the present invention, the glycosidic group is a disaccharide modified to bear at least one hydrophobic substituent. In a preferred embodiment of the present invention, each of the groups A2, A4, A5, A6 and A7 bears an aromatic side chain and the aromatic side chains of groups A2 and A6 are linked to the aromatic side chain of group A4 via ether linkages and the aromatic side chains of groups A5 and A7 are linked to each other via a carbon-carbon bond. In another preferred embodiment of the invention, the group A4 bears a glycosidic group. The glycosidic group is preferably is a disaccharide comprising a glucose residue directly bonded to group A4 and a vancosamine residue bonded to said glucose residue. In preferred compounds of the present invention, A2-A3-A4-A5-A6-A7 is as found in a compound selected from the group consisting of vancomycin, eremomycin, chloroeremomycin, and β-avoparcin, more preferably A2-A3-A4-A5-A6-A7 is as found in vancomycin. In other preferred compounds of the present invention, the C6 position of the glucose residue attached to A4 is modified to bear at least one substituent other than hydroxyl, and more preferably the at least one substituent other than hydroxyl is a polar substituent or a hydrophobic substituent. In preferred compounds of the present invention where A2-A3-A4-A5-A6-A7 is as found in vancomycin, the vancosamine residue in vancomycin is N-substituted with said at least one hydrophobic substituent. In other preferred compounds of the present invention wherein A2-A3-A4-A5-A6-A7 is as found in vancomycin, the glucose residue attached to A4 of vancomycin is modified at the C6 position to bear at least one substituent other than hydroxyl, preferably a polar substituent, and said vancosamine residue is N-substituted with said at least one hydrophobic substituent. The at least one hydrophobic substituent is preferably selected from R, OR, NR1R, SR, SO2R, C(O)OR, C(O)SR, C(S)OR, C(S)SR, NR1C(O)R, C(O)NR1R, or halo and R is alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl or arylsulfonyl; R1 is hydrogen, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl or arylsulfonyl; and any pharmaceutically acceptable salts thereof; and if two or more of said substituents are present, they can be the same or different. The organic group A1 in the preferred compounds of the present invention, is preferably an organic group selected from the group consisting of a modified or unmodified alpha amino acid residue other than N-substituted leucine, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl, arylsulfonyl, guanidinyl, carbamoyl, or xanthyl. Also, in the preferred compounds of the present invention, the A7 group bears a terminal carboxyl, ester, thioester, amide, N-substituted amide, or other carboxylic acid derivative, any of which groups may be substituted with any of the substituents described herein.

[0034] In another aspect, the present invention is also directed to a method for making compounds of the formula A1-A2-A3-A4-A5-A6-A7 wherein each of the groups A2 to A7 comprises a modified or unmodified α-amino acid residue, A1 comprises an organic group other than N-substituted leucine, and at least one of the groups A1 to A7 is linked via a glycosidic bond to one or more glycosidic groups each having one or more sugar residues, wherein at least one of said sugar residues is modified to bear at least one hydrophobic substituent comprising the steps of removing the N-substituted leucine residue from the compound N-substituted-leucyl-A2-A3-A4-A5-A6-A7 thereby forming a compound having a free amino group at A2; and attaching an organic group A1 to the free amino group at A2, wherein the hydrophobic substituent and the groups A1-A7 are as described above. Preferably, the N-substituted-leucine residue is N-methyl leucine. This method is applicable to any of the preferred compounds as described above.

[0035] In another aspect, the present invention is directed to a method for making a glycopeptide antibiotic having the formula A1-A2-A3-A4-A5-A6-A7 wherein A2-A3-A4-A5-A6-A7 is as found in vancomycin and A1 comprises an organic group other than N-substituted leucine, said method comprising modifying vancomycin to form a modified vancomycin bearing a hydrophobic substituent at the vancosamine nitrogen; removing the N-methyl leucine residue from the modified vancomycin to form a des-N-methyl leucyl modified vancomycin bearing a free amino group at A2 and attaching an organic group A1 to the amino group at A2, wherein the hydrophobic substituent and the organic group A1 are as defined above.

[0036] In another aspect, the present invention is directed to a method for making a glycopeptide antibiotic having the formula A1-A2-A3-A4-A5-A6-A7 wherein A2-A3-A4-A5-A6-A7 is as found in vancomycin and A1 comprises an organic group other than N-substituted leucine, said method comprising modifying vancomycin to form a first modified vancomycin bearing a substituent other than hydroxyl at the C6 position of the glucose attached to A4 of vancomycin; modifying said first modified vancomycin to form a second modified vancomycin bearing a hydrophobic substituent at the vancosamine nitrogen; removing the N-methyl leucine residue from said second modified vancomycin to form a des-N-methyl leucyl second modified vancomycin bearing a free amino group at A2; and, attaching an organic group A1 to the amino group at A2, wherein the hydrophobic substituent and the organic group A1 are as described above. It is preferred in this method that the substituent other than hydroxyl at the C6 position of the glucose attached to A4 of vancomycin is a polar substituent.

[0037] The present invention is also directed to a method of treating an infectious disease in a host comprising administering to said host an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or ester thereof. Preferably the host is a mammalian host, more preferably a human. The infectious disease is preferably a bacterial infection. The present invention is also directed to a composition comprising a compound of the present invention or a pharmaceutically acceptable salt or ester thereof and a pharmaceutically acceptable carrier or excipient. The compound of the present invention may be administered solely or in combination with any other drug or therapeutic agent.

[0038] The present invention is also directed to a method for removing an N-terminal amino acid residue from an oligopeptide or polypeptide comprising reacting an oligopeptide or a polypeptide with phenylisothiocyanate in a pyridine-water-triethylamine solvent medium. Preferably, the reaction of the oligopeptide or polypeptide with phenylisothiocyanate is carried out in a 10:10:1 (volume) pyridine-water-triethylamine solvent medium. The reaction is preferably carried out at a temperature in the range of from 40-70° C. and for a period of time in the range of from 20-60 minutes. In preferred methods, the N-terminal amino acid residue is N-methyl leucine. The preferred oligopeptides are selected from the group consisting of a glycopeptide antibiotic, a pseudoaglycone and an aglycone, preferably a glycopeptide antibiotic or pseudoaglycone in which at least one of the glycosidic groups therein is modified to bear at least one hydrophobic substituent. Preferably, the glycopeptide antibiotic is vancomycin. More preferably, the disaccharide at A4 of vancomycin is modified to bear at least one hydrophobic group. Preferably, the vancosamine nitrogen at A4 of vancomycin is modified to bear at least one hydrophobic group. In preferred methods, the glucose residue attached directly to A4 of vancomycin is modified to bear at least one substituent other than hydroxyl, which is preferably a polar substituent or a hydrophobic substituent. Where the glucose residue is modified to bear at least one substituent other than hydroxyl, it is preferred that the C6 position of the glucose residue attached directly to A4 of vancomycin is modified to bear a polar or a hydrophobic substituent.

DETAILED DESCRIPTION OF THE INVENTION

[0039] Herein we disclose strategies for finding promising glycopeptide compounds with good activity against sensitive and resistant bacterial strains. One strategy involves attaching substituents to the free amino group of amino acid 2 in des-N-substituted-leucine glycopeptides and analogs thereof containing at least one hydrophobic substituent on a glycosidic group attached to one of the amino acids A2-A7. We have also discovered that where at least one of the glycosidic groups attached to one of the amino acids A2-A7 bears a hydrophobic substituent, it is not necessary to attach a group to the free amino group of amino acid A2 upon removal of the N-substituted leucine residue in order to produce compounds having biological activity. Thus, we have found good activity in des-N-substituted leucine glycopeptide compounds in which at least one of the glycosidic groups attached to one of the amino acids A2-A7 bears a hydrophobic substituent and in which A2 bears a free amino group upon removal of the N-substituted leucine residue therefrom. We have demonstrated the utility of the strategy by making a set of compounds, of which several have better activity against a range of strains than the corresponding compounds in which A1 is N-substituted leucine, which is preferably N-methyl leucine. Some of the substitutions improve activity against both sensitive and resistant strains relative to N-methyl leucine; others improve activity more against sensitive strains than resistant strains; still other improve activity more against resistant strains than sensitive strains. Thus, it is possible to manipulate the biological activity in different ways by choosing appropriate A1 substituents. It is also possible to manipulate the biological activity by choosing appropriate hydrophobic substituents. We also show that the physical properties of the compounds—e.g., the hydrophobic-hydrophilic balance as measured by HPLC retention times—are related to both the hydrophobic substituent, and when present, the specific group A1. Having shown that both the physical properties and the biological activities of glycopeptide derivatives containing hydrophobic substituents on the sugars are affected by the identity of the hydrophobic substituent and, when present, the group A1, the present invention is thus directed to all compounds of the general structure A1-A2-A3-A4-A5-A6-A7 wherein the group A1 is optional and, when present, is preferably an organic group having from 2-30 carbon atoms, which may contain heteroatoms. The organic group A1 may contain more than 30 carbon atoms. The organic group A1, when present, is attached to the amino group at A2. The organic group A1 may be linear, branched or cyclic, or some combination thereof, and may include aliphatic, aromatic, and/or heterocyclic groups, provided that it is not a leucine or a modified leucine residue, and provided that it is not directly or indirectly linked by a covalent bond to amino acid 3. Preferably the organic group is a modified or unmodified alpha amino acid residue, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl, arylsulfonyl, guanidinyl, carbamoyl, or xanthyl.

[0040] When present, preferred A1 groups are provided by the compounds listed in Table I following Example 3. The structural formulae of these preferred A1 groups are also provided in Table I. These are merely preferred A1 groups, and the present invention is not to be construed as being limited thereto. In general, A1 can be any organic group that can be reacted with the free amino group at A2 by, for example, formation of an amide linkage. Thus, preferred reagents which can be reacted with the free amino group at A2 include compounds having a carboxylic acid group which reacts with the free amino group at A2 to form the amide linkage. Such preferred compounds include those disclosed in Table I.

[0041] In preferred compounds of the present invention, each of the groups A2 to A7 comprises a modified of unmodified α-amino acid residue, whereby (i) the group A1,when present, is linked to an amino group on the group A2, (ii) each of the groups A2, A4 and A6 bears an aromatic side chain, which aromatic side chains are cross-linked together by two or more covalent bonds, and (iii) the group A7 bears a terminal carboxyl, ester, thioester, amide, N-substituted amide, or other derivative of a carboxylic acid.

[0042] In the compounds of the present invention, one or more of the groups A2 to A7 is linked via a glycosidic bond to one or more sugar resides; wherein at least one of said sugar resides bears at least one hydrophobic substituent wherein the hydrophobic substituent is preferably selected from R, OR, NR1R, SR, SO2R, C(O)OR, C(O)SR, C(S)OR, C(S)SR, NR1C(O)R, C(O)NR1R, or halo and R is alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl or arylsulfonyl; R1 is hydrogen, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl or arylsulfonyl; and any pharmaceutically acceptable salts thereof; and if two or more of said substituents are present, they can be the same or different.

[0043] Modified amino acid residues include amino acid residues whose aromatic groups have been substituted by halo, alkyl, alkoxy, alkanoyl, or other groups easily introduced by electrophilic substitution reactions or by reaction of phenolic hydroxyl groups with alkylating or acylating agents; and amino acid residues which have protecting groups or other easily introduced substituents on their hydroxyl or amino groups including, but not limited to alkyl, alkanoyl, aroyl, aralkyl, aralkanoyl, carbamoyl, allyloxycarbonyl, aralkyloxycarbonyl, aryloxycarbonyl, alkylsulfonyl, arylsulfonyl, heterocyclic, heterocyclic-alkyl or heterocyclic-carbonyl substituents. Examples of preferred protecting groups include acetyl, allyloxycarbonyl (aloc), CBZ, allyl, benzyl, p-methoxybenzyl and methyl. Modifications of hydroxyl groups occur on phenolic hydroxyl groups, benzylic hydroxyl groups, or aliphatic hydroxyl groups. Other amino acid residues, in addition to A2, A4 and A6 may be cross-linked through their aromatic acid substituent groups.

[0044] In the preferred compounds of the present invention, the residues A2 to A7 of the glycopeptides are linked sequentially by peptide bonds and are cross-linked as in a dalbaheptide. The preferred glycopeptides have a peptide core in which the residues are linked as in the glycopeptide antibiotics vancomycin, eremomycin, chloroeremomycin or β-avoparcin. In particularly preferred compounds of the present invention, the structures and interconnections of A2 to A7 are those of vancomycin, i.e., those having the heptapeptide core of vancomycin with the N-methyl leucine residue removed, subject to the amino acid modifications and substitutions described herein above.

[0045] The glycopeptide compounds of the present invention contain at least one glycosidic group attached through a glycosidic bond to at least one of the amino acid residues A2 to A7. In the preferred compounds of the present invention, a glycosidic group is linked to residue A4. This glycosidic group comprises at least a monosaccharide bearing at least one hydrophobic substituent. Preferably, the glycosidic group is a disaccharide residue bearing at least one hydrophobic substituent which disaccharide residue can be linked to any of the amino acid residues A2-A7, preferably to the amino acid residue A4. In the particularly preferred compounds of the present invention, the glycosidic group attached to A4 is a disaccharide consisting of a glucose residue directly attached to the amino acid A4 and an N-substituted vancosamine residue attached to the glucose residue. Preferably, the vancosamine residue is N-substituted with the at least one hydrophobic substituent. Examples of preferred hydrophobic substituents which are preferably present as N-substituents on the vancosamine residues are shown below:

[0046] Thus, preferred N-substituents at the vancosamine nitrogen include, e.g., straight or branched alkyl, aralkyl, alkanoyl, aralkanoyl and aroyl. Any of these N-substituents may be substituted with one or more of alkyl, preferably C4-C8 alkyl, halo, haloalkyl, aryl, aralkyl, aryloxy, aralkyloxy, alkaryloxy, alkoxy, preferably C4-C8 alkoxy, and haloalkoxy. Preferred alkoxy substituents on the N-substituent include, e.g., O-butyl and O-octyl. Where the N-substituent is alkyl, it is preferred that alkyl has from 8 to 15 carbon atoms. Specific preferred N-subsitutents include, but are not limited to the following:

[0047] 2-naphthylmethyl

[0048] 4-phenylbenzyl

[0049] 1-naphthylmethyl

[0050] 4-phenoxybenzyl;

[0051] 4-benzyloxybenzyl

[0052] 4-trifluoromethoxybenzyl

[0053] 4-allyloxybenzyl

[0054] 4-nonyloxybenzyl;

[0055] 2-methoxy-1-naphthylmethyl

[0056] 4-dodecyloxybenzyl

[0057] 9-phenanthranylmethyl

[0058] 4-decyloxybenzyl

[0059] 9-anthranylmethyl

[0060] 4-[phenylethynyl]4-phenylbenzyl

[0061] 4-methoxy-1-naphthylmethyl

[0062] 1-pyrenylmethyl

[0063] 9-[10-methyl]anthranylmethyl

[0064] 9-[10-chloro]anthranylmethyl

[0065] 2-benzthienylmethyl

[0066] 4-[4-hydroxyphenyl]benzyl

[0067] 4-[4-octylphenyl]benzyl

[0068] 4-[4-pentylphenyl]benzyl

[0069] 4-[4-octyloxyphenyl]benzyl

[0070] 3-pyridylmethyl

[0071] 5-nitro-1-naphthylmethyl

[0072] 4-pyridylmethyl

[0073] 4-quinolylmethyl

[0074] 3-quinolylmethyl

[0075] 4-stilbenzyl

[0076] 2-quinolylmethyl

[0077] 2-pyridylmethyl

[0078] 2-fluorenylmethyl

[0079] 4-phenoxyphenethyl

[0080] 4-[4-pentylcyclohexyl]benzyl

[0081] 4-benzylphenethyl

[0082] 4-[4-biphenyl]benzyl

[0083] 4-trifluoromethylbenzyl

[0084] trans-cinnamyl

[0085] 4-[1-oxa]fluorenylmethyl

[0086] 4-[4-pentoxyphenyl]benzyl

[0087] 4-thiomethylbenzyl

[0088] 2,3-[2-methyl-3→4-t-butylphenyl]]propenyl

[0089] 9-(1-methyl)-acridinylmethyl

[0090] 2-hydroxy-1-naphthylmethyl

[0091] 4-[2-phenyl-6-methoxy]quinoylmethyl

[0092] 4-diphenylmethylbenzyl

[0093] 3,4 cyclohexenylmethyl

[0094] 3,4-methylenedioxylbenzyl

[0095] 3-phenoxybenzyl

[0096] 4-benzylbenzyl

[0097] 3-benzyloxy-6-methoxy benzyl

[0098] 4-benzyloxy-3-methoxybenzyl

[0099] 3,4-dibenzyloxybenzyl

[0100] 4-[4-methoxyphenyl]benzyl

[0101] 4-[3-cyanopropoxy]benzyl

[0102] 3,4-ethylenedioxybenzyl

[0103] 4-[4-nitrophenoxy]benzyl

[0104] 2,3-methylenedioxybenzyl

[0105] 2-benzyloxyphenethyl

[0106] 2-ethoxy-1-naphthylmethyl

[0107] 2-benzylfurylmethyl

[0108] 3-phenoxyphenethyl

[0109] 4-phenoxyphenethyl

[0110] 4-[4-nitrophenyl]benzyl

[0111] 6-methoxy-2-naphthylmethyl

[0112] 3-methyl-5-thienylmethyl

[0113] 5-phenyl-2-thienylmethyl

[0114] 4-benzyloxyphenethyl

[0115] 3-benzyloxyphenethyl

[0116] 4-[2-nitrophenoxy]benzyl

[0117] 5-[4-methoxyphenyl]-2-thienylinethyl

[0118] 4-difluormethoxybenzyl

[0119] 2,3,4,5,6-pentamethylbenzyl

[0120] 5-iodo-2-thienylmethyl

[0121] 4-[2-[2-chloroethoxy]ethoxy]benzyl

[0122] 3,4-dimethylbenzyl

[0123] 3-acetoxybenzyl

[0124] 4-nitrobenzyl

[0125] 4-phenylethynylbenzyl

[0126] 4-[2-chloro-6-fluorobenzyloxy]benzyl

[0127] 4-[3,4-dichlorophenoxy]benzyl

[0128] 4-[3,4-dichlorobenzyloxy]benzyl

[0129] S-[2,3-dihydrobenzfuryl]methyl

[0130] 4-[2-[N,N-diethylamino]ethoxy]benzyl

[0131] 2-bicyclo[2.1.2]heptylmethyl

[0132] 2-hydroxy-5-phenylbenzyl

[0133] 3-[4-chlorophenoxy]benzyl

[0134] 4-[3-chlorophenoxy]-3-nitrobenzyl

[0135] 4-[2-chlorophenoxy]-3-nitrobenzyl

[0136] 3,5-dimethylbenzyl

[0137] 4-[4-ethylphenyl]benzyl

[0138] 3-phenylbenzyl

[0139] 4-[3-fluorophenyl]benzyl

[0140] 4-[4-chlorobenzyloxy]benzyl

[0141] 4-[4-chlorophenoxy]-3-nitrobenzyl

[0142] 4-[4-methylphenoxy]benzyl

[0143] 4-[4-t-butylphenoxy]benzyl

[0144] 4-[4-methylphenyl]benzyl

[0145] 4-[4-methoxyphenoxy]benzyl

[0146] 4-acetoxy-3-methoxybenzyl

[0147] 4-[(2-phenyl)ethyl]benzyl

[0148] 3-[5-phenyl]pyridinylmethyl

[0149] 4-[2-nitrophenyl]benzyl

[0150] 2-[1-hydroxy]fluorenylmethyl

[0151] 4-benzyl-3-methoxybenzyl

[0152] 4-[cyclohexylmethoxy]-3-ethoxybenzyl

[0153] 3-[3,3′-dichlorophenoxy]benzyl

[0154] 4-[4-propylphenyl]benzyl

[0155] 4-thiophenylbenzyl

[0156] 4-[alpha-hydroxybenzyl]benzyl

[0157] 2,2-dinitro-4-thiophenebenzyl

[0158] 3-[3-trifluoromethylphenoxy]benzyl

[0159] 4-[t-butylethynyl]benzyl

[0160] 4-phenoxy-3-methoxy-benzyl

[0161] 4-[3-trifluoromethylphenoxy]-3-nitrobenzyl

[0162] 2-phenylthiobenzyl

[0163] 2-[4-chlorophenyl]-6-benzoxazolemethyl

[0164] 4-[alpha-methoxybenzyl]benzyl

[0165] 4-cyclohexylbenzyl

[0166] 3-[3,4-dichlorophenoxy]benzyl

[0167] acenaphthlenylmethyl

[0168] 4-[1,1,2,2-tetrafluoroethoxy]benzyl

[0169] 4-benzoyloxy-3,3-dimethoxybenzyl

[0170] 3-[cyclohexylmethoxy]benzyl

[0171] 4-cyclohexyloxybenzyl

[0172] 3-[2-quinoylmethoxy]benzyl

[0173] 4-[alpha-ethoxybenzyl]benzyl

[0174] 4-[cyclohexylethoxy]benzyl

[0175] 4-[alpha-propoxybenzyl]benzyl

[0176] 4-[4-methyl-1-piperidino]benzyl

[0177] 2-thiophene-1,2-cyclohexenylmethyl

[0178] 4-[4-nitrobenzyloxy]benzyl

[0179] 3-[4-trifluoromethylphenoxy]benzyl

[0180] 3-benzoyl-2,4-dichlorobenzyl

[0181] 4-[2-[2-thiopropyl]ethoxy]benzyl

[0182] 4-[2-methyl-1-piperidino]benzyl

[0183] 4-hydroxybenzyl

[0184] 4-[2-pyridyl]benzyl

[0185] 4-acetoxybenzyl

[0186] 5,6-benzonorbomylmethyl

[0187] 3-phenylcyclopentylmethyl

[0188] 1-adamantylmethyl

[0189] 3-[cyclohexylmethoxy]-4-methoxybenzyl

[0190] 2-[2-glucosyl]benzyl

[0191] 4-[4-pentoxybiphenyl]benzyl

[0192] 3,4-dihydroxybenzyl

[0193] 4-[4-methylpiperazino]benzyl

[0194] 4-morpholinobenzyl

[0195] 4-[4-chlorophenylsulfonyl]benzyl

[0196] 4-methylsulfonyloxybenzyl

[0197] 4-benzoyloxybenzyl

[0198] 5-phenyl-3-pyridinylmethyl

[0199] 4-[N,N-bis(2-chloroethyl)amino]benzyl

[0200] 3-cyclohexyloxybenzyl

[0201] 4-[2-t-butoxyethoxy]benzyl

[0202] 3,3-dichloro-4-hydroxy-benzyl

[0203] 1,2,3,4,-tetrahydro-9-anthranylmethyl

[0204] 4-cyclohexanoyloxybenzyl

[0205] 4-nonanoyloxybenzyl

[0206] 4-[phenylsulfinyl]benzyl

[0207] 4-anilinobenzyl

[0208] cyclohexylmethyl

[0209] 3-benzoyloxybenzyl

[0210] 3-nonanoyloxybenzyl

[0211] 4-[cyclohexyl]cyclohexylmethyl

[0212] 3-cyclohexanoyloxybenzyl

[0213] 4-[cyclohexanoyloxy]-3,3→dimethoxy]benzyl

[0214] 4-[nonanoyloxy]-3,3→dimethoxy]benzyl

[0215] 1,2,3,4-tetrahydro-6-naphthylmethyl

[0216] 2-hydroxybenzyl

[0217] [2-[6,6-dimethyl-bicyclo>3.1.1]hept-2-enyl]methyl

[0218] 1-cyclohexenyl-4-isopropylmethyl

[0219] 4-[4-methoxyphenyl]butyl

[0220] 4-[[2,3,4,5,6-pentamethyl]phenylsulfonyloxy]benzyl

[0221] 4-[1-pyrrolidinosulfonyl]benzyl

[0222] 3-[4-methoxyphenyl]propyl

[0223] 8-phenyloctyl

[0224] 4-[2,3-dihydroxypropoxy]benzyl

[0225] 4-[N-methylanilino]benzyl

[0226] 2-[2-napthyl]ethyl

[0227] 6-methyl-2-naphthyl methyl

[0228] cis-bicyclo[3.3.0]octane-2-methyl

[0229] 2-tridecynyl

[0230] 4-butyl-2-cyclohexylmethyl

[0231] 4-[(4-fluorobenzoyl)amino]benzyl

[0232] 4-[(3-fluorobenzoyl)amino]benzyl

[0233] 8-phenoxyoctyl

[0234] 6-phenylhexyl

[0235] 10-phenyldecyl

[0236] 8-bromooctyl

[0237] 11-tridecynyl

[0238] 8-[4-methoxyphenoxy]octyl

[0239] 8-[4-phenylphenoxy]octyl

[0240] 8-[4-phenoxyphenoxy]octyl

[0241] 3-[3-trifluoromethylphenoxy]benzyl

[0242] 10-undecenyl

[0243] 4-cyclohexylbutyl

[0244] 4-phenyl-2-fluorobenzyl

[0245] 7-hexadecynyl

[0246] 3-[cyclopentyl]propyl

[0247] 4-[2-methylphenyl]benzyl

[0248] 4-[phenylazo]benzyl

[0249] 4-[4-flurophenyl]benzyl

[0250] 3-nitro-4-[4-nitrophenyl]benzyl

[0251] 3-nitro-4-[2-nitrophenyl]benzyl

[0252] 9-decenyl

[0253] 4-[3,4-dimethoxyphenyl]benzyl

[0254] 4-[4-trifluromethylphenyl]benzyl

[0255] 5-hexenyl

[0256] 4-[2-thienyl]benzyl

[0257] 4-[6-phenylhexyloxy]benzyl

[0258] 9,10-dihydro-2-phenantrene methyl

[0259] 4-[3,4-dimethylphenyl]benzyl

[0260] 4-[4-methylphenyl]-2-methylbenzyl

[0261] 4-[3-phenylpropyloxy]benzyl

[0262] 4-[3-methylphenyl]benzyl

[0263] 4-[4-methylphenyl]-3-methylbenzyl

[0264] 4-[4-pentenyloxy]benzyl

[0265] 4-[1-heptynyl]benzyl

[0266] 3-[4-t-butyl-phenylthio]benzyl

[0267] 4-[4-chlorophenyl]benzyl

[0268] 4-[4-bromophenyl]benzyl

[0269] 4-[4-cyanophenyl]benzyl

[0270] 4-[1-nonynyl]benzyl

[0271] 4-[11-tridecynyloxy]benzyl

[0272] 12-phenyldodecyl

[0273] 6-phenyl-5-hexynyl

[0274] 11-phenyl-10-undecynyl

[0275] 4-[2-methylphenyl]-3-methylbenzyl

[0276] 3-[2-thienyl]-2-thienylmethyl

[0277] 4-[benzyloxymethyl]cyclohexylmethyl

[0278] 4-[4-chlorophenoxy]benzyl

[0279] 4-[benzyl]cyclohexylmethyl

[0280] 4-benzoylbenzyl

[0281] 4-[phenoxymethyl]benzyl

[0282] 4-[4-chlorobenzyl]benzyl

[0283] In another preferred embodiment of the present invention, the glucose residue attached to A4 is modified to bear a substituent, which may be any of the hydrophobic substituents as described above as well as polar substituents. Thus, the preferred compounds of the present invention encompass compounds in which the vancosamine residue is N-substituted with a hydrophobic substituent and the glucose residue is modified to bear a substituent other than hydroxyl. Preferably, it is the C6 position of the glucose residue that is modified to bear a substituent other than hydroxyl as described above. Thus, in particularly preferred compounds of the present invention, the vancosamine residue is N-substituted with a hydrophobic substituent and the glucose residue is also modified at the C6 position to bear a substituent other than hydroxyl. Where the vancosamine residue is N-substituted with a hydrophobic substituent and the glucose residue is also modified to bear a substituent other than hydroxyl, it is preferred that glucose residue attached to A4 is substituted with a polar substituent. Examples of preferred substituents on the glucose residue, and in particular at the C6 position of the glucose residue include, but are not limited to, mesitylenesulfonyl; 2-thio-6-azathymine; 2-thio-4-hydroxy-6-methylpyrimidine; 2-thio-5-amino-1,3,4-thiadiazole; 2-thio-4-amino-3-hydrazino-1,2,4-triazole; 2-thio-4-hydroxy-6-methylpyrimidine; 2-thio-6-azathymine; iodo; amino; azido; bromo; hydrazino; iminotriphenylphosphoranyl; S-3-amino-5-mercapto-1,2,4-triazolyl; N-2-quinoxalinyl-Vancosamine. It is to be understood that the C6 position can also be modified to bear any of the hydrophobic substituents as described above. Thus, where the C6 position of the glucose residue attached directly to A4 is modified to bear a hydrophobic substituent as described above, it is not necessary that the vancosamine residue attached to the glucose residue also bear a hydrophobic substituent as well. However, it is possible that both the glucose and vancosamine glycosidic groups at the A4 position can be modified to bear a hydrophobic substituent.

[0284] The invention is not intended to be limited to the embodiments described above. Thus, beneficial effects of at least some of the A1 substituent replacements on a dalbaheptide in which A1 is not covalently linked to A3 would be expected to apply generally to glycopeptide derivatives containing at least one hydrophobic group on a sugar covalently bonded to the peptide.

[0285] The compounds of the present invention can be prepared by removing the terminal N-substituted leucine residue from a compound of the formula N-substituted leucyl-A2-A3-A4-A5-A6-A7 wherein at least one of the glycosidic groups attached to any of A2-A7 bears a hydrophobic substituent to form the compound des-N-substituted-leucyl-A2-A3-A4-A5-A6-A7 bearing a free amino group at A2 and then attaching the group A1 to the free amino group at A2 to form a compound of the present invention. The N-substituted leucine residue is preferably N-methyl leucine. The terminal N-substituted leucine residue can be removed by any conventional process for removing a terminal amino acid from an oligopeptide or polypeptide. One conventional method to remove a terminal amino acid is Edman degradation. This method is described in the literature and can be readily employed to remove a terminal N-substituted leucine residue in a process of making the compounds of the present invention. As applied to the method of forming the des-N-methyl leucyl compounds of the present invention, Edman degradation involves the reaction of the amino group of the terminal N-methyl leucine residue with phenyl isothiocyanate in a suitable solvent. An intermediate thiourea compound is formed, and the N-methyl leucine residue splits off from the thiourea as a phenylthiohydantoin, resulting in the corresponding des-N-methyl leucine compound.

[0286] Thus, the N-methyl leucine residue can be removed from the compound N-methyl leucyl-A2-A3-A4-A5-A6-A7 by reacting this compound with phenylisothiocyanate in a suitable organic solvent, preferably a pyridine-water solvent, more preferably, a 1:1 pyridine-water solvent at a temperature of about 50° C. This reaction generates the corresponding thiourea, which is then treated with TFA-CH2Cl2 to yield the des-N-methyl leucyl compound, to which the group A1 can optionally be attached as described in more detail below.

[0287] The present inventors have also discovered a modified Edman degradation procedure by which an N-terminal amino acid residue can be removed from a polypeptide or an oligopeptide. This procedure involves reacting the N-terminal amino acid residue on the polypeptide or oligopeptide with phenylisothiocyanate in a pyridine-water-triethylamine solvent medium. The pyridine-water-triethylamine solvent medium preferably comprises pyridine-water-triethylamine in a ratio of 10:10:1 by volume. The reaction is conducted for about 20 to 60 minutes, with 60 minutes preferred. While not wishing to be bound by any particular theory, it is believed that triethylamine is a key reagent in this modified Edman degradation protocol. It is believed that the triethylamine catalyzes the in situ conversion of the initially formed thiourea to the final product.

[0288] In the context of the present invention, the modified Edman degradation process described above can be applied to remove the terminal N-substituted leucine residue from the compound N-substituted leucyl-A2-A3-A4-A5-A6-A7 wherein at least one glycosidic group attached to any of A2-A7 bears a hydrophobic substituent to yield the corresponding desleucyl compound bearing a free amino group at A2 which can then be reacted with the organic group A1 to yield the compounds of the present invention. Thus, the compound N-substituted leucyl-A2-A3-A4-A5-A6-A7, wherein the N-substituted leucine residue is preferably N-methyl leucine, is reacted with phenylisothiocyanate in a pyridine-water-triethylamine solvent medium, preferably a 10:10:1 pyridine-water-triethylamine solvent medium and at a temperature of about 50° C. This method yields the corresponding des-N-methyl leucyl compound in one step in nearly quantitative yield. The desired des-N-methyl leucine compound can be precipitated from DMF by adding excess of 20% ethyl acetate-hexane. The resulting product is then suitably pure for a subsequent optional step of attaching the group A1 to the free amino group at A2 on the des-N-methyl leucine compound to form a compound of the present invention.

[0289] Any conventional method can be employed to attach the group A1 to the free amino group at A2 after removal of the terminal N-substituted leucine residue. Such methods of coupling amino groups to other organic groups are well known to the ordinarily skilled chemist. In the preferred compounds of the present invention the group A1 is attached to the free amino group at A2 by forming an amide linkage. Thus, a carboxylic acid or other amine-reactive compound can be reacted with the free amino group at A2 to form preferred compounds of the present invention. It is also possible to attach the organic group to the free amino group at A2 by reductive alkylation or other common methods of functionalizing amino groups. It may be desirable in some cases when attaching the group A1 to the free amino group at A2 to suitably protect free amino groups at other positions in the intermediate compound so as to selectively attach the A1 group to the free amino group at A2. Such methods of selectively protecting free amino groups and selectively removing the protective groups are well known to the ordinarily skilled chemist. Suitable protecting groups for free amino groups include 9-fluorenylmethyloxycarbonyl (Fmoc), benzyloxycarbonyl (CBZ), tert-butyloxycarbonyl (t-Boc), and allyloxycarbonyl (alloc).

[0290] Preferably, the terminal N-substituted leucine residue is removed from a dalbaheptide wherein at least one of the groups A2-A7 is linked via a glycosidic bond to one or more glycosidic groups each having one or more sugar residues and wherein at least one of the sugar residues is modified to bear at least one hydrophobic substituent as described above. Thus, for example, N-methyl leucine can be removed from the dalbaheptide vancomycin in which the vancosamine residue is N-substituted with a hydrophobic substituent and which optionally may also be substituted on the glucose residue, preferably at the C6 position thereof, with either a hydrophobic or, more preferably, a polar substituent as described above. Also, N-methyl leucine can be removed from the dalbaheptide vancomycin in which the C6 position is modified to bear a hydrophobic substituent and in which the vancosamine nitrogen optionally also bears a hydrophobic substituent. The modified des-N methyl leucyl vancomycin can then be reacted to attach the A1 substituent to form the compounds of the present invention.

[0291] An N-substituted vancomycin glycopeptide can be prepared by attaching a hydrophobic substituent to the amino group on the vancosamine residue by reductive alkylation or other conventional methods for functionalizing an amino group. Thus for example, an aldehyde can be reacted with vancomycin in a suitable organic solvent, followed by reduction of the aldehyde carbonyl group with a suitable reducing agent followed by conventional separation and purification, which may involve recrystallization and/or reverse phase chromatographic techniques as are well known to the ordinarily skilled chemist. In some cases it may be desirable to selectively protect the amino group in the N-methyl leucine residue prior to the reductive alkylation of the vancosamine nitrogen. Any amino protecting group may be employed and conventional methods of removing the amino protecting group may be employed to remove the protective group after performing the reductive alkylation at the vancosamine nitrogen. Other methods of coupling free amino groups to organic substituents can also be employed to attach the hydrophobic substituent the vancosamine amino group. Thus, reductive alkylation is merely a preferred method of attaching the hydrophobic group to this position of the vancosamine residue, and other methods will be apparent to the person having ordinary skill in the art.

[0292] As discussed above, in addition to modifying a glycosidic group to bear at least one hydrophobic substituent as described above in connection with the vancosamine nitrogen, it may also be desirable to modify another glycosidic group to bear a hydrophobic or polar substituent. In fact, any glycosidic group attached to any of the amino acid residues A2-A7 can be modified to bear a hydrophobic substituent in accordance with the present invention. Thus, any of the glycosidic groups in, e.g., the glycopeptide antibiotics vancomycin, eremomycin, chloroeremomycin, and β-avoparcin can be modified to bear at least one hydrophobic substituent. Thus, the present invention is not to be construed as limited to the preferred compounds which comprise a hydrophobic substituent at the vancosamine nitrogen and, optionally, a substituent at the C6 position of the glucose residue directly attached to amino acid A4 in vancomycin.

[0293] It is to be understood that where vancomycin is modified to bear a substituent at the C6 position of the glucose residue directly attached to A4, this substituent may be hydrophobic or polar, however, it is preferred that where the vancosamine nitrogen position is modified to bear a hydrophobic substituent, the C6 position, when modified, will preferably be modified to bear a polar substituent. Where the C6 position of glucose attached to A4 of vancomycin is to be modified to bear a substituent, the following strategy may be employed to introduce a suitable set of protecting groups onto vancomycin and to differentiate the C6 hydroxyl group of the glucose residue on A4 of vancomycin from all other hydroxyl groups. This strategy may be generally employed to modify any selected hydroxyl group of a glycopeptide antibiotic which need not be limited to vancomycin, although C6 modification of the glucose residue attached to A4 of vancomycin is preferred. Thus, the following method is particularly suitable for modification of, e.g., a primary hydroxyl group on any glycosidic group attached to any of the amino acid residues A2-A7. Thus, while the foregoing method is described in reference to the primary hydroxyl group at the C6 position of the glucose residue directly attached to A4 of vancomycin, it is to be understood that the synthetic method described below is applicable to any similarly reactive hydroxyl group on any glycosidic group attached to any of the amino acid residues A2-A7. A schematic which generally illustrates the modification of the C6 position of vancomycin is shown below:

[0294] Protection of both amines by similar groups requires using excess acylation reagent while selective protection of the N-methyl leucine residue is known, allowing selective functionalization of the vancosamine amine group. See Pavlov et al., J. Antibiotics, 1993, 46, 1731, incorporated herein in its entirety. Selectively introducing the mesitylenesulfonyl group at the glucose C6 position differentiates this position from the other hydroxyl groups and allows further reaction to displace the mesitylenesulfonyl group, affording may derivatives. A variety of functional groups are introduced at the glucose C6 position by using common methods for nucleophilic displacement of primary arylsulfonyl groups directly, or by further synthetic modification of initial displacement products, including azido and iodo groups. For example, the iodo group is displaced by a variety of nucleophiles to produce additional C6 derivatives. A preferred nucleophile is a thiol compound, especially a heterocyclic thiol. Modification of an azido group at the C6 position is performed, e.g., by reducing the azido group to an amino group, which in turn is functionalized by means of reductive alkylation, nucleophilic substitution, or other amino-group reactions well known to those skilled in the art. In a preferred embodiment of the invention, an azido group is partially reduced by reaction with a phosphine compound to produce an iminophosphorane. In a preferred method of modifying the C6 position, or other similarly reactive hydroxyl group on a glycosidic group, the C6 position is modified to bear a free amino group by displacing the mesitylenesulfonyl group with an azido group which is then reduced to the free amino group. The free amino group at the C6 position can then be further modified to bear, e.g., a hydrophobic substituent by reacting the free amino group in a manner similar to that described above with respect to attaching a hydrophobic substituent to the vancosamine nitrogen.

[0295] In the process described above, the vancosamine amino group, the N-methyl leucine amino group and the carboxyl group at A7 of vancomycin are suitably protected. Then, the C6 hydroxyl is substituted with a mesitylenesulfonyl group which, as described above can be further displaced, e.g., by nucleophilic displacement to afford many derivatives. While the above method has been described in connection with attachment of mesitylenesulfonyl group at the C6 position, it is to be understood that after suitably protecting the glycopeptide starting compound, the C6 hydroxyl group can be reacted with any compound that will attach a good leaving group to the C6 position. The leaving group may then be displaced by a subsequent reaction, e.g., by nucleophilic displacement, and further derivatization may then be performed at the C6 position yielding many derivatives. The groups protecting the vancosamine amino group, the N-methyl leucine amino group and the carboxyl group at A7 can be removed in a conventional manner.

[0296] Where the C6-substituted vancomycin analog is to be further substituted with a hydrophobic substituent at the vancosamine nitrogen, the protecting groups at the vancosamine amino group, at the carboxyl group at A7 and at the N-methyl lecuine amino group of the C6-substituted vancomycin are removed. The vancosamine nitrogen is then substituted with the hydrophobic substituent in the manner described above, e.g., by performing a reductive alkylation reaction. The N-methyl leucine residue is removed from this compound and the group A1 is attached to the free amino group at A2 as described above. The resulting compound, may then be further modified at the C6 position as described above in connection with the C6 derivatization, thus affording many derivatives. The resultant product, after separation and purification will thus have a hydrophobic substituent at the vancosamine nitrogen position and will have been modified to bear a group other than hydroxyl at the C6 position of the glucose residue, and will also have an organic group other than N-methyl leucine attached to the amino acid A2.

[0297] Preferably, the N-substituted leucine residue is removed from a compound in which one or more of the glycosidic groups attached to one of the amino acid residues A2-A7 is already modified to bear the at least one hydrophobic substituent. However, it is also possible to modify the glycosidic group either prior to or after removal of the N-substituted leucine residue. Thus, it is possible to attach one or more glycosidic groups onto a glycopeptide antibiotic, pseudoaglycone or aglycone bearing a terminal N-substituted leucine residue and then modifying the glycosidic group to bear the at least one hydrophobic substituent. Furthermore, the modification of the glycosidic group can also be conducted either prior to or after attachment of the A1 group upon removal of the terminal N-substituted leucine residue.

[0298] The glycosidic group can be attached to any reactive hydroxyl group in a glycopeptide antibiotic, aglycone or pseudoaglycone. Preferably the glycosidic group is attached to an aglycone or to a pseudoaglycone. Where the glycosidic group is attached to an aglycone, it is preferable to attach a second glycosidic group to the previously attached glycosidic group, which results in a disaccharide group attached to one of the amino acid residues in the aglycone. Preferably, the sequential attachment of glycosidic groups is performed at the A4 position of an aglycone, however, it is to be understood that this method can be generally applied to any of the amino acid residues forming an aglycone, pseudoaglycone or glycopeptide antibiotic. The glycosidic groups can be attached to any of the reactive hydroxyl groups in glycopeptide antibiotics, aglycones or pseudoaglycones. These reactive hydroxyl groups are generally phenolic hydroxyl groups, benzylic hydroxyl groups or aliphatic hydroxyl groups. Thus, a glycosidic group can be introduced at any of such hydroxyl groups as desired. Moreover, as discussed above, a glycosidic group can also be attached to a previously attached glycosidic group, which results in a disaccharide group attached to one of the amino acid residues in the aglycone, pseudoaglycone or glycopeptide antibiotic. Any hydroxyl group on the glycopeptide antibiotic, aglycone or pseudoaglycone to which a glycosidic group is not desired to be attached can be suitably protected. Also, the glycosidic group itself may be suitably protected so that the desired glycosidic bond to the glycopeptide antibiotic, aglycone or pseudoaglycone is formed.

[0299] Thus, a glycopeptide antibiotic having a terminal N-substituted leucine residue can be prepared by (a) selecting: (i) an aglycone that is soluble in one or more organic solvents, is derived from a glycopeptide antibiotic, and which aglycone has exactly one free phenolic hydroxyl group; and (ii) a protected first glycosyl donor; (b) allowing a non-enzymatic glycosylation reaction to proceed in an organic solvent such that a first glycosidic bond is formed, which links said free phenolic hydroxyl group to the anomeric carbon of the first glycosyl donor to provide a pseudoaglycone having a protected first glycosyl residue; (c) selectively removing one protecting group from the first glycosyl residue to provide a pseudoaglycone bearing exactly one free hydroxyl group on the first glycosyl residue; (d) selecting a second protected glycosyl donor; and (e) allowing a non-enzymatic glycosylation reaction to proceed in an organic solvent such that a second glycosidic bond is formed which links said free hydroxyl group on the pseudoaglycone to the anomeric carbon of the second glycosyl donor. Any of the glycosidic groups on the resultant compound can be modified to bear the at least one hydrophobic substituent in accordance with the preferred methods as described above. Any glycosidic group can be attached to an aglycone, pseudoaglycone or glycopeptide antibiotic in the foregoing manner. Thus, it may be desirable to attach a glycosidic group bearing a free amino group to an aglycone, pseudoaglycone or glycopeptide antibiotic as described above. A hydrophobic substituent can then be attached to the free amino group to produce a compound having a glycosidic group bearing at least one hydrophobic substituent in accordance with the present invention. Attachment of a glycosidic group with a free amino group is advantageous because it may avoid the necessity of functionalizing the glycosidic group to bear an amino group prior to attaching the hydrophobic substituent thereto.

[0300] It is apparent that the method described above can be modified by starting with a pseudoaglycone and then attaching another glycosidic group thereto. Thus it is to be understood that the method of suitably protecting and deprotecting hydroxyl groups can be generally applied to selectively attach a glycosidic group to any desired hydroxyl group on an aglycone, pseudoaglycone or glycopeptide antibiotic, any of which may be modified to bear a hydrophobic substituent on a glycosidic group.

[0301] Where it is desired to attach glycosidic groups to an aglycone, pseudoaglycone or glycopeptide antibiotic, all reactive functional groups on any of these starting compounds are suitably protected. Thus, amine, carboxylic acid, phenolic and benzylic hydroxyl groups, e.g., need to be protected to avoid their participation in the reaction that attaches the glycosidic group. The protecting groups are suitably chosen so as to render the protected compounds soluble in the reaction medium. The protecting groups may remain on the final compound, but are preferably removed by exposure to acidic or basic conditions, catalytic hydrogenation, or light or other conventional methods for removing protecting groups. Any conventional protecting groups for the functional groups mentioned above may be employed. When the aglycone, pseudoaglycone or glycopeptide antibiotic is or is derived from vancomycin, it is preferred that the protecting groups are as follows: carboxybenzyl (CBz) on the amino nitrogen, a benzyl ester group; benzyl, allyl or methyl phenolic ethers on the phenolic hydroxyls of A5 and A7 , and acetates on the aliphatic hydroxyls. Removal of the protective groups can be accomplished by methods well known to the ordinarily skilled organic chemist. Thus, when it is desired to remove protecting groups from any of the compounds of this invention, their removal is accomplished using methods well known to those skilled in the art. The preferred method for removal of protecting groups is as follows. Aloc groups on amines, and allyl esters or allyl ethers are removed by using Pd(O) mediated reactions, e.g., [Ph3P]2Pd(II)Cl2 and Bu3SnH in 1:1 acetic acid:DMF. Acetate protecting groups are removed using hydrazine in THF/methanol. The use of protecting groups to protect any group which might otherwise be reactive under a particular set of reaction conditions is well known to the ordinarily skilled artisan. As will be apparent to the ordinarily skilled artisant, any such conventional protecting groups and the methodologies employed therewith can be used in the present invention.

[0302] The suitably protected aglycone, pseudoaglycone or glycopeptide antibiotic is glycosylated via a non-enzymatic reaction in an organic solvent with a variety of glycosyl donors, thereby forming a glycosidic bond between the aglycone, pseudoaglycone, glycopeptide antibiotic and the glycosyl donor. Preferably, the glycosyl donors are activated monosaccharide anomeric sulfoxides which are fuctionalized at the 6 position or elsewhere. These sulfoxide donors are differentially protected so as to allow for selective deprotection of a single hydroxyl after formation of the glycosidic bond. The single hydroxyl can then be the reactive site for forming another glycosidic bond with a glycosidic group. Suitable protecting groups to allow for this selective deprotection include, but are not limited to, the 2,2-dimethyl acetoacetate group, the 4-azidobutyryl group and any other groups which can be removed in the presence of other protecting groups.

[0303] Glycosidic groups can also be removed from a glycopeptide antibiotic or pseudoaglycone. Thus, a glycosidic group can be removed from a glycopeptide antibiotic by (a) selecting a glycopeptide antibiotic that is soluble in one or more organic solvents; (b) contacting the glycopeptide antibiotic with a Lewis acid, and allowing a degradation reaction to proceed such that a sugar residue is removed, producing a pseudoaglycone having exactly one free hydroxyl group on a remaining sugar residue of the pseudoaglycone; a glycosidic group can then optionally be attached to the free hydroxyl group on the pseudoaglycone by the subsequent steps of (c) selecting a protected glycosyl donor; and (d) allowing a non-enzymatic glycosylation reaction to proceed in an organic solvent such that a glycosidic bond is formed which links the free hydroxyl group of the remaining sugar residue on the pseudoaglycone to the anomeric carbon of the glycosyl donor. Thus, the foregoing method can be applied to removal of a glycosidic group, e.g., from a glycopeptide antibiotic having a disaccharide attached to A4. The glycopeptide antibiotic bearing a disaccharide at A4 is treated with a Lewis acid in an organic solvent to remove a sugar residue from the disaccharide group. The Lewis acid is preferably boron trifluoride, preferably as a complex with diethyl ether. When the glycopeptide antibiotic having a disaccharide group at A4 is vancomycin, it is preferred that allyloxycarbonyl (aloc) groups are present on the amines of A1 and the vancosamine residue, acetates on the aliphatic hydroxyl groups, allyl phenyl ethers on the phenolic hydroxyls, and an allyl or o-nitrobenzyl ester on the A7 terminal carboxyl, where a solid-phase synthesis is employed, the o-nitrobenzyl ester is preferred. A degradation reaction then proceeds which remove a glycosidic group to produce a pseudoaglycone in which all reactive functional groups (amine, carboxylic acid, phenols, and benzylic alcohols) are suitably protected except for a hydroxyl group on the remaining glycosidic group attached to residue A4 which is where another glycosidic group can optionally be attached.

[0304] Pharmaceutical formulations of the compounds of the present invention are also a part of the present invention, as well as the use of the compounds and formulations thereof to treat infectious diseases in mammals, preferably humans, comprising administering an amount of the compound of the present invention or a pharmaceutically acceptable salt or ester thereof to a mammal, the amount being effective to treat the infectious disease.

[0305] Thus, the compounds of the present invention, or pharmaceutically acceptable salts or esters thereof can be formulated for any conventional means of delivery, including oral or parenteral delivery for the therapeutical or prophylactic treatment of infectious diseases, preferably bacterial diseases. The bacterial diseases which may be therapeutically or prophylactically treated with the compounds and/or formulations of the present invention include those caused by Gram-positive and/or Gram-negative microorganisms.

[0306] The compounds of the present invention may be administered separately or in combination with any other drug or therapeutic agent. Examples of other therapeutic agents and/or drugs that can be administered with the compounds and/or formulations of the present invention include, but are not limited to, beta lactam antibiotics, such as penems, penams, cephems, carbapenems, oxacephems, carbacephems, and monobactams, or other antibiotics such as cycloserine and fosfomycin. The other therapeutic agent need not be an antibiotic.

[0307] The compounds and/or formulations are administered to the mammal in a therapeutically effective amount, which amount is effective to treat, prevent, mitigate and/or alleviate the infectious disease. Thus, the compound of the present invention can be administered to the mammal, preferably a human, in an amount ranging from about 0.5 to about 2 grams per day. The compounds and/or formulations of the present invention can be administered in a single daily dosage or in multiple doses per day. Other periodic treatment protocols may also be adopted. Thus, the treatment protocol may require administration over extended periods of time, e.g., for several days or for from about one to six weeks. The therapeutically effective amounts of the compound of the invention discussed above are merely exemplary. Thus, the amount per administered dose or the total amount administered will depend on such factors as the nature and severity of the infection, the age and general health of the patient, the tolerance of the patient to the compounds and/or formulations of the present invention and the microorganism or microorganisms involved in the infection.

[0308] In the pharmaceutical formulations of the present invention, the compound can be admixed with any conventional pharmaceutical carriers and/or excipients and can be formulated for immediate or sustained release. Other time-release profiles, such as combinations of immediate and sustained release are also possible. Thus, the compound of the present invention can be admixed with conventional pharmaceutical carriers and excipients and used in the form of tablets, capsules, caplets, elixirs, suspensions, syrups, wafers and the like. The compounds of the present invention can also be formulated for topical administration. Typical excipients and/or carriers include, but are not limited to corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, and alginic acid. Disintegrators commonly used in the formulations of this invention include croscarmellose sodium, microcrystalline cellulose, corn starch, sodium starch glycolate and alginic acid. Tablet binders that can be included are acacia, methylcellulose, polyvinylpyrrolidone, hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose. Lubricants that can be used include magnesium stearate or other metallic stearates, stearic acid, silicone fluid, talc, waxes, oils and colloidal silica. Flavoring agents such as peppermint, oil of wintergreen, cherry flavoring or the like can also be used. It may also be desirable to add a coloring agent to make the dosage form more esthetic in appearance or to help identify the product. Tablets may be coated to facilitate swallowing or to modify release of the active compound, or some combination of these.

[0309] The compounds and/or formulations can also be administered intravenously or intramuscularly. For intravenous (IV) use, a water-soluble form of the compound is preferably dissolved in one of the commonly used intravenous fluids, and administered by infusion. Such fluids as, for example, physiological saline, Ringer's solution or 5% dextrose can be used. For intramuscular preparations, a sterile formulation of a suitable salt or ester form of the compound of the present invention, for example the hydrochloride salt form can be dissolved and administered in a pharmaceutical diluent such as water-for-injection, physiological saline or 5% glucose solution. A suitable insoluble form of the compound may be prepared and administered as a suspension in an aqueous base or a pharmaceutically acceptable oil base, such as an ester of a long chain fatty acid such as ethyl oleate.

[0310] For oral use, a sterile formulation of a suitable salt or ester form of the compound, for example, the hydrochloric acid salt, formulated in a diluent such as distilled or deionized water is particularly useful. Alternatively, the unit dosage form of the compound can be a solution of the compound, preferably in its salt or ester from, in a suitable diluent in sterile, hermetically sealed ampoules.

EXAMPLES

[0311] The present invention will now be described with reference to the specific examples below, to which the present invention is not to be construed as limited to.

Example 1

p-chlorobiphenyl vancomycin

[0312] The structural formula of p-chlorobiphenyl vancomycin is shown below:

[0313] To a solution of vancomycin hydrochloride (20 mg; 13 μmoles) in 1.5 mL DMF was added diisopropylethylamine (11 μL, 65 μmoles) and 4,4′-chlorobiphenylaldehyde (280 μL of a 10 mg/mL solution in DMF; 13 μmoles). The reaction mixture was stirred at 60° C. for half an hour. Sodium cyanoborohydride (77 μL of a 0.5M solution in DMF) was added, and the system was stirred at 60° C. for 1 hour.

[0314] The reaction mixture was cooled to room temperature and diluted with 25 mL of ethyl ether. The precipitate was collected and purified by reverse phase HPLC:

[0315] HPLC Conditions for Product Analysis:

1Column:Phenomenex C18 column, 21.2 × 250 mmFlow:8 mL/minMobile Phase:B: acetonitrile4: 10 mM ammonium acetate, pH 5.2Program: 0 min 30% B 0.1 min 30% B 25 min 55% B (linear gradient) 35 min 90% B (linear gradient)35.5 min 90% B (linear gradient) 45 min 30% B

Example 2

Des-leucyl p-chlorobiphenyl vancomycin (TS-518)

[0316] The structural formula of des-leucyl-p-chlorobiphenyl vancomycin is shown below:

[0317] Under an argon atmosphere, p-chlorobiphenyl vancomycin (130 mg, 7.8 μmoles) was dissolved in 3.4 mL of a 10:10:1 mixture of freshly distilled pyridine, distilled water and triethylamine (99%). Sonication was used to promoted total dissolution. To the colorless solution was added phenyl thioisocyanate (11 μL, 90.1 μmoles), and the system was kept at 50° C. for 20 minutes. The slightly yellow solution was transferred to a separatory funnel, diluted with 10 mL of 10:10:1 pyridine-water-triethylamine solution, and washed with 10 mLof 10% ethyl acetate-hexane. The top layer was discarded. The yellowish bottom layer was transferred to a round-bottom flask, diluted with 5 mL of 2-butanol and concentrated to dryness. The residue was azeotroped twice with toluene. The resulting solid was dissolved in minimal amount of DMF (3 mL) and the product was precipitated by adding a large volume of 50% ethyl acetate-hexane (40 mL). The precipitate was collected by filtration, washed with methylene chloride (3×10 mL) and dried under vacuum to afford a nearly quantitative yield of des-leucyl p-chlorobiphenyl vancomycin (off-white solid). The product may be further purified by flash chromatography on silica gel (3:3:2 ethyl acetate-ethanol-water).

[0318] HPLC Conditions for Product Analysis:

2Column:Nucleosil 4 C18 100 A (250 × 4.6 mm)Flow:0.75 mL/minMobile Phase:B: acetonitrile5: 10 mM ammonium acetate, pH 5.2Program: 0 min 25% B 0.1 min 25% B 20 min 40% B (linear gradient) 30 min 90% B (linear gradient)30.1 min 25% B (linear gradient) 40 min 25% BRetention time11.6 min.of product:

Example 3

[0319] Compounds were typically prepared in batches of 48. To each of 48 test tubes was added the appropriate carboxylic acid (0.77 mmoles). Bis-(6-carboxy-HOBT)-N-(2-aminoethyl)-aminomethyl polystyrene resin (1.56 mmole/g; purchased from NovaBiochem; 1.19 g) was suspended in 28 ml of amino-free DMF using mild, yet thorough, stirring. An aliquot of the suspension (500 μl) was added to each test tube, followed by 500 μL of amino-free DMF and 100 uL of a solution of 1,3-diisopropyl-carbodiimide in DMF (prepared by adding 750 μoL of 1,3-diisopropyl-carbodiimide to 5 mL of amine-free DMF). The test tubes were shaken on an orbital shaker at 350 rpm for one hour. The supernatant was removed by filtration and discarded. The resin was washed with 2 mL of amine-free DMF (6×).

[0320] Des-(N-methyl-leucyl)-p-chlorobiphenyl-vancomycin (1.5 g) was dissolved in 50 mL of amine-Free DMF using sonication. An aliquot of this solution (1 mL) was added to each test tube, followed by 1 mL of amine-Free DMF. The test tubes were shaken on an orbital shaker at 350 rpm for one hour. The supernatant of each reaction mixture was transferred to the well of a labeled 48-well plate. The resin was washed with 1 mL of amine-free DMF (2×) and the washings were combined with the supernatant. In the cases where the carboxylic acid contained an Fmoc group, a 20% solution of piperidine in DMF (1 mL) was added to the corresponding well.

[0321] The plates were then dried in a centrifugal evaporator. The residues were treated with 5 mL of DMSO (molecular biology grade) and sonicated until total dissolution. The resulting solutions were used as such for analytical analysis and biological screening.

[0322] In the structural formulae in Table I below, “X” designates the —COOH group in the compound that is reacted with the free amino group at A2, forming the new amide linkage.

[0323] The antibacterial activity of each of the compounds against the bacterial strains E. faceium (ATCC 49624), S. epidermidis (ATCC 12228), S. aureus (ATCC 29213), E. faecalis (CL 4877) and E. faecalis (ATCC 292121) was tested. Each of the compounds was screened in a 96 will agar array format. Antibacterial activity was referenced to the zones of inhibition observed for p-chlorobiphenylvancomycin. The MIC's of p-chlorobiphenylvancomycin against resistant isolates were approximately 6 ug/ml. A zone score of 2 was assigned when the zone of inhibition for a given compound was equal to the zone generated by the delivery of a 1 mg/ml stock solution of p-chlorobiphenylvancomycin. A zone score of 1 was assigned if the area of the zone was 25% of the area of the zone generated by a 1 mg/ml stock solution of p-chlorobiphenylvancomycin. Likewise a zone score of 3 was assigned if the zone size was 4 times the size of the zone generated by a 1 mg/ml stock solution of p-chlorobiphenylvancomycin. Similarly, a zone score of 4 was assigned if the zone size was 16 times the size of the zone generated by a 1 mg/ml stock solution of p-chlorobiphenylvancomycin and a zone score of 5 was assigned if the zone size was 64 times the size of the zone generated by a 1 mg/ml stock solution of p-chlorobiphenylvancomycin. The screening data for each of the compounds is presented in Table I, below.

3TABLE ISide Chains attached to Free Amino Group at A2 of des-leucyl-p-chlorobiphenyl vancomycin and Biological Screening ResultsE. faeciumS. epidermidisS. aureusE. faecalisE. faecalisCmpd.ATCCATCCATCCCLATCCNo.Reagent Name4962412228292134877292121

ALA244412

ASN(TRT)111213

ASP(OTBU)132214

CHA132215

CIS233216

CYC(MMT)111217

PHE132218

SAR344229

SER(TRT)1111110

THI2323111

THR(TRT)1112112

TRP1224113

CYS(TRT)1111114

D-CYS(TRT)1112115

D-MET4552416

D-PHE3543317

D-SER(TBU)5555518

2323119

TYR(TBU)1323120

VAL2434121

L-(+)-LACTIC ACID1222122

MYR-GLY1223123

(R,S)-2-CARBOXY MORPHOLINE1222124

4-PIPERAZIN-1-YL ACETIC ACID2234125

D-TRP2422226

GLU(OBZL)1322127

GLN(TRT1113128

GLY2432229

HIS(TRT)1111130

HYP(TBU)2435131

(3S,4S)-4-AMINO- 3-HYDROXY-6- METHYLTHIO- HEXANOIC ACID1221132

3-AMINO-1- CARBOXY METHYL- PYRIDIN-2-ONE2322133

4-(2- AMINOETHYL)-1- CARBOXY- METHYL- PIPERAZINE DIHYDRO CHLORIDE2322134

2-CARBOXY METHYL- PIPERAZINE2345135

ALA-ALA1222136

ALA-GLY2445137

ILE1335138

LEU2421139

LYS(DDE)1322140

LYS(MTT)1112141

MET2432142

NLE2445143

ALLO-THR2342144

ASN(GLCNAC(AC) 3-BETA-D)1111145

ASN(TMOB)1222146

BETA, BETA- DIMETHYL-D- CYS(ACM)4552447

BETA-ALA2432148

CYS(2-HYDROXY ETHYL)1222149

CYS(ACM)1422150

CYS(ME)2412151

D-ALA4552452

D-ALLO-THR3441153

D-ASN1242154

D-CIS-HYP1222155

D-CYS(ACM)3442156

D-DPR(DDE)2433157

D-GLN2433258

D-HIS1112159

D-ISO ASPARAGINE1222160

DL-ISOSER1112161

D-LYS (CARBAMYL)4553462

D-ORN (CARBAMYL)4544463

D-SER3422164

D-THR4542265

GAMMA-ABU1222166

GLN(TMOB)1222167

GLY-GLY-GLY1224168

GLY-GLY2335169

GLY-PRO-HYP1112170

GLY-VAL1112171

HIS1112172

HYP2232173

L-ASPARAGINE1111174

L-ISO ASPARAGINE1221175

L-LYS(BIOTIN)2212276

LYS(AC)2211177

LYS(BIOTINYL- EPSILON- AMINOCAPROYL)1115178

LYS(CARBAMYL)1221179

LYS(FOR)2325180

LYS(ME)3 CHLORIDE2425181

MET(O)2235182

MET(O2)2334183

ORN(PYRAZINLY CARBONYL)2324184

PEN(ACM2221185

PRO-GLY2222186

PRO-PRO1222187

SER(AC)1222188

SER2222189

THR2212190

N-ALPA-L- ARGININE1122191

N-ALPHA-L- GLUTAMINE1235192

N-ALPHA-N-BETA- ALOC-L-DIAMINO PROPIONIC ACID1332193

N-ALPH-N- GAMMA-ALLOC- L-DIAMINO BUTYRIC ACID1223194

(2-CARBOXY ETHYL) DIMETHYL SULFONIUM CHLORIDE1222195

(3-ACETYL-2- METHYL-5-OXO-2- ACETIC ACID1113196

(S)-(−)-4-OXO-2- AZETIDINE CARBOXYLIC ACID1112197

[3-METHYOXY CARBONYL)-2- METHYL-5-OXO-2- PYRROLIN-4- YL]ACETIC ACID1111198

100

1-(AMINO CARBONYL)-1- CYCLOPROPANE CARBOXYLIC ACID1122199

101

1-ACETYL PIPERIDINE-4- CARBOXYLIC ACID12311100

102

2-(2-METHOXY ETHOXY)ACETIC ACID11231101

103

2-[2-(2-METHOXY ETHOXY) ETHOXY]ACETIC ACID11121102

104

2-ACETAMIDO ACRYLIC ACID11121103

105

2-PYRAZINE CARBOXYLIC ACID11121104

106

3,4-DIACETAMIDO BENZOIC ACID11221105

107

3-AMINO PYRAZINE-2- CARBOXYLIC ACID11121106

108

3-HYDROXY PROPIONIC ACID12221107

109

4-ACETAMIDO BUTYRIC ACID11221108

110

4-(NITRO BENZOYL- GLYCYL- GLYCINE11241109

111

5-AMINOOROTIC ACID11211110

112

AC-ALA11211111

113

AC-ARG11121112

114

AC-D-ALA11241113

115

AC-D-ASN11121114

116

AC-DL-LYS(AC)11121115

117

AC-D-MET11221116

118

AC-D-PRO12221117

119

ACETOXYACETIC ACID12211118

120

ACETYL-DL- CARNITINE HYDROCHLORIDE12241119

121

ACETYL-L- CARNITINE- HYDROCHLORIDE12221120

122

AC-GLY-GLY12221121

123

AC-HYP11221122

124

AC-LEU-GLY11111123

125

AC-LYS(AC)11211124

126

AC-MET(O)11241125

127

AC-THR11121126

128

ALLANTOIC ACID11121127

129

ARABIC ACID11121128

130

ARABINIC ACID11211129

131

BETAINE HYDROCHLORIDE12121130

132

BICINE11221131

133

BOC-ALA-GLY- GLY11221132

134

BOC-ALA-GLY- SAR11221133

135

BOC-ASN11121134

136

BOC-ASP-NH212321135

137

BOC-D-ASN12231136

138

BOC-D-GLN12141137

139

BOC-GLU-NH211221138

140

BOC-GLY-ARG11121139

141

BOC-GLY-GLY- GLY11221140

142

BOC-GLY-GLY11121141

143

BOC-L-GLUTAMINE11221142

144

BOC-MET(O)21221143

145

BOC-MET(O2)11211144

146

CACOTHELINE11121145

147

CREATINE11121146

148

D-(−)-QUINIC ACID11121147

149

D-(+)- GALACTURONIC ACID MONOHYDRATE11121148

150

D-ALPHA- GALACTURONIC ACID11121149

151

D-CARNITINE HYDROCHLORIDE21121150

152

D-GLUCURONIC ACID11111151

153

DL-CARNITINE HYDROCHLORIDE22421152

154

DL-GLYCERIC ACID11121153

155

DL-PYRO GLUTAMIC ACID11111154

156

D-PYRO GLUTAMIC ACID11111155

157

D-SACCHARIC ACID 1,4- LACTONE11121156

158

D-SACCHARIC ACID 3,5- LACTONE11111157

159

GLUCONIC ACID11221158

160

GLYCOLIC ACID12221159

161

GLYOXYLIC ACID11121160

162

GUANIDOACETIC ACID11111161

163

HIPPURYL-GLY- GLY-OH11121162

164

HYDANTOIC ACID11121163

165

HYDANTOIN-5- ACETIC ACID11211164

166

LACTOBIONIC ACID11111165

167

L-ARGININIC ACID11121166

168

L-BETA-IMIDAZO LELACTIC ACID11111167

169

L-CARNITINE HYDROCHLORIDE11111168

170

L-DIHYDRO OROTIC ACID11121169

171

L-PYRO GLUTAMIC ACID11121170

172

MALEAMIC ACID12121171

173

METHANE SULFONYL ACETIC ACID11121172

174

N-(ACETO ACETYL) GLYCINE11121173

175

N,N-DIMETHYL SUCCINAMIC ACID11111174

176

N-ACETYL-DL- ALANINE11121175

177

N-ACETYL-DL- METHIONINE12211176

178

N-ACETYL-DL- PROLINE12221177

179

N-ACETYL-DL- PROPARGYL- GLYCINE11221178

180

N-ACETYL-DL SERINE11121179

181

N-ACETYL GLYCINE11121180

182

N-ACETYL-L- GLUTAMINE11111181

183

N-ACETYL-L- HISTIDINE12221182

184

N-ACETYL-L- METHIONINE12121183

185

N-ACETYL-L- PROLINE12221184

186

N-ALPHA- ACETYL-L- ARGININE DIHYDRATE11121185

187

N-ALPHA- ACETYL-L- ASPARAGINE11111186

188

BETA-GUANIDINO PROPIONIC ACID11111187

189

N-ALPHA- CARBAMYL-L- ARGININE11221188

190

N-ALPHA- CARBOETHOXY- L-ASPARAGINE11221189

191

N-CARBOMOYL MALEAMIC ACID11221190

192

N-CARBAMYL- ALPHA-AMINO- ISOBUTYRIC ACID11231191

193

N-CARBAMYL-DL- ALPHA-AMINO-N- BUTYRIC ACID11131192

194

N-CARBAMYL-L- HISTIDINE HYDROCHLORIDE11121193

195

N-FORMYL GLYCINE12321194

196

N-FORMYL-L- ALANINE11221195

197

N-FORMYL-L- HISTIDINE12221196

198

N-FORMYL-L- METHIONINE11231197

199

NICOTINURIC ACID11221198

200

OROTIC ACID12231199

201

OXALYL MONOGUANYL HYDRAZIDE11221200

202

OXAMIC ACID11221201

203

SHIKIMIC ACID11221202

204

SUCCINAMIC ACID12221203

205

SUCCINIC ACID 2,2-DIMETHYL HYDRAZIDE11211204

206

SUCCINIC SEMIALDEHYDE11221205

207

SULFOACETIC ACID11221206

208

DL-2-UREIDO PROPIONIC ACID12221207

209

THYMINE-1- ACETIC ACID12431208

210

URACIL-5- CARBOXYLIC ACID11221209

211

Z-ALA-GLY-GLY12221210

212

Z-BETA-ALA-GLY- GLY12221211

213

Z-GLN-GLY12221212

214

Z-GLY-GLN12221213

215

Z-GLY-GLY-ALA12221214

216

Z-GLY-GLY-GLY- GLY (SEQ ID NO. 1)11221215

217

Z-GLY-GLY-GLY11221216

218

DELTA-VAL11211217

219

D-ARG23321218

220

ACPC11211219

221

DELTA-ABU11221220

222

ALA24411221

223

3-UREIDO PROPIONIC ACID11221222

224

CARBOXY METHOXYL AMINE HEMIHYDRO CHLORIDE11221223

225

BOC-GLN-GLN-OH11221224

226

(−)-2-OXO-4 THIAZOLIDINE- CARBOXYLIC ACID12221225

227

(ETHYLTHIO) ACETIC ACID13421226

228

(METHYLTHIO) ACETIC ACID12411227

229

(R)-(−)-5-OXO-2- TETRAHYDRO FURAN CARBOXYLIC ACID12211228

230

1H-TETRAZOLE-1- ACETIC ACID12121229

231

1-METHYL PYRROLE-2- CARBOXYLIC ACID12221230

232

2-(4-CHLORO PHENYLTHIO) NICOTINIC ACID11221231

233

2-(TRIFLUORO METHYL) PROPENOIC ACID11221232

234

2,2-BIS(HYDROXY METHYL) PROPIONIC ACID11221233

235

2,4,5-TRICHLORO PHENOXY ACETIC ACID11221234

236

2,4-DICHLORO PHENYLACETIC ACID12221235

237

2,4-DIHYDROXY BENZOIC ACID11111236

238

2,4-DIMETHOXY BENZOIC ACID11221237

239

2-AMINO NICOTINIC ACID12421238

240

2-FLUORO BENZOIC ACID12211239

241

2-FURAN GLYOXYLIC ACID11211240

242

2-FUROIC ACID12211241

243

2-HYDROXY NICOTINIC ACID11121242

244

2-KETOBUTYRIC ACID11121243

245

2-METHYL PYRAZINE-5- CARBOXYLIC ACID11221244

246

3-(2-FURYL) ACRYLIC ACID12231245

247

3-(3,4- DIMETHOXYL PHENYL) PROPIONIC ACID12221246

248

3-(PHENYL SULFONYL) PROPIONIC ACID24531247

249

3-(TRIFLUORO METHYL) BENZOIC ACID12321248

250

3-(TRIFLUORO METHYLTHIO) BENZOIC ACID11221249

251

3,3,3-TRIFLUORO PROPIONIC ACID13421250

252

3,4,5- TRIACETOXY BENZOIC ACID12311251

253

3,4-DICHLORO CINNAMIC ACID11121252

254

3,4-DIFLUORO PHENYLACETIC ACID12411253

255

3,4-DIMETHOXY BENZOIC ACID12221254

256

3,5-BIS (TRIFLUORO METHYL) BENZOIC ACID11221255

257

3,5-DIHYDROXY BENZOIC ACID11221256

258

3-AMINO-1,2,4- TRIAZOLE-5- CARBOXYLIC ACID11221257

259

3-AMINOBENZOIC ACID12221258

260

3-ETHOXY PROPIONIC ACID12221259

261

3-FLUORO BENZOIC ACID12311260

262

3-FUROIC ACID12311261

263

3-HYDROXY BENZOIC ACID12221262

264

3-HYDROXY BUTYRIC ACID12221263

265

3-METHYL THIOPROPIONIC ACID14511264

266

4-(METHYL SULFONYL) BENZOIC ACID12201265

267

4-ACETAMIDO BENZOIC ACID11221266

268

4-ACETOXY BENZOIC ACID12221267

269

4-ACETYL BENZOIC ACID22411268

270

4-AMINO BENZOIC ACID11121269

271

4-CARBOXY BENZENE SULFONAMIDE12221270

272

4-FLUORO BENZOIC ACID12221271

273

4-HYDROXY BENZOIC ACID12241272

274

4-IMIDAZOLE CARBOXYLIC ACID12221273

275

4-METHOXY CINNAMIC ACID12331274

276

4-AMINO SALICYLIC ACID12221275

277

5-METHYL ISOXAZOLE-4- CARBOXYLIC ACID12221276

278

6-HYDROXY NICOTINIC ACID12311277

279

6-HYDROXY PICOLINIC ACID12321278

280

6-OXOHEPTANOIC ACID11221279

281

ACETIC ACID12321280

282

ANTHRANILIC ACID12321281

283

BENZOTRIAZOLE- 5-CARBOXYLIC ACID12221282

284

COUMALIC ACID11221283

285

PYRAZINE-2,3- DICARBOXYLIC ACID MONOAMIDE12241284

286

D-DESTHIO BIOTIN12321285

287

DIFLUOROACETIC ACID12221286

288

DL-2-HYDROXY- N-BUTYRIC ACID12221287

289

ETHOXY ACETIC ACID12221288

290

FUMARIC ACID MONETHYL ESTER12421289

291

GLYOXYLIC ACID SEMICARBAZONE12211290

292

HEPTAFLUORO BUTYRIC ACID12221291

293

INDOLE-2- CARBOXYLIC ACID12221292

294

ISONICOTINIC ACID12421293

295

ITACONIC ACID MONOMETHYL ESTER12221294

296

LACTIC ACID12321295

297

LEVULINIC ACID12421296

298

MALEIC ACID MONOETHYL ESTER12321297

299

MALEIC ACID MONOMETHYL ESTER11221298

300

2,3-DIHYDRO-3- OXOPYRIDAZINE- 6-CARBOXYLIC ACID11221299

301

MAYBRIDGE BTB 0931612241300

302

MAYBRIDGE KM 0150212321301

303

MAYBRIDGE KM 0600012221302

304

3-(4-CHLORO BENZENE SULPHONYL) BUTYRIC ACID13431303

305

METHOXY ACETIC ACID12441304

306

MONO-METHYL GLUTARATE12431305

307

MONO-METHYL SUCCINATE12431306

308

N-[3-(2-FURYL ACRYLOYL)]- GLYCINE12321307

309

NICOTINIC ACID13421308

310

NICOTINIC ACID N-OXIDE12421309

311

N0METHYL MALEAMIC ACID12321310

312

PENTAFLUORO PHENYLACETIC ACID12421311

313

PERFLUORO PENTANOIC ACID11211312

314

PICOLINIC ACID12221313

315

PICOLINIC ACID N-OXIDE12221314

316

PYRUVIC ACID12221315

317

SALICYLIC ACID11121316

318

TETRAHYDRO-2- FUROIC ACID12231317

319

TETRAHYDRO-3- FUROIC ACID13421318

320

THIOPHENE-2- ACETIC ACID12431319

321

THIOPHENE-2- CARBOXYLIC ACID12321320

322

THIOPHENE-3- ACETIC ACID12321321

323

UROCANIC ACID12221322

324

HSE(ME)12221323

325

L-THREONINE MONOHYDRATE12221324

326

N-ACETYL-DL- HISTIDINE HYDRATE12221325

327

N-FORMYL-DL- ALANINE12321326

328

N-FORMYL-DL- METHIONINE12221327

329

OXAMIC ACID12331

[0324] In Table II, below, MIC (minimum inhibitory concentration) values of certain compounds of the present invention are provided for the bacterial strains E. faecium ATCC 49624, E. faecium CL 4931, E. faecalis ATCC 29212, E. Faecalis CL 4877, S aureus ATCC 29213, and S. Aureus ATCC 33591. The minimum inhibitory concentrations (MIC) of test compounds were determined using bacteria grown in brain heart infusion media (BHI) supplemented with 0.1% casamino acids. Logarithmically growing cells were diluted to approximately 5×105 CFU/ml and subjected to test compounds solubilized and serially diluted in DMSO. A 5% final DMSO concentration had no affect on cell viability or killing. After 18 hours at 37° C., the OD600 was determined by reading the ninety-six well microtiter plates on a microplate reader. For a given concentration, an MIC determination was made if:

[OD600 Control−OD600 Test Conc.]/[OD600 Control−OD600 Media]×100≧90%

4

TABLE II

MIC Values of Compounds of the Invention Against Selected Baterial Strains

330

E. faecium

E. faecalis

S. aureus

Cmpd

ATCC
CL
ATCC
CL
ATCC
ATCC

No.
R1
R2
Reagent Name
49624
4931
29212
4877
29213
33591

0
OH

331

Vancomycin (Vancosamine sugar is unsubstituted in this compound only.)
1.25
>250
3.12
>250
1.25
2.5

1
OH

332

L-ALA
0.78
25
1.56
25
0.78
0.78

7
OH

333

L-PHE
0.12
6.25
0.78
6.25
0.39
0.39

11
OH

334

L-THR(TRT)
3.12
6.25
3.12
3.12
3.12
3.12

12
OH

335

L-TRP
0.39
6.25
1.56
3.12
0.78
0.78

15
OH

336

D-MET
0.12
12.5
0.25
12.5
0.25
0.12

16
OH

337

D-PHE
0.12
12.5
0.25
12.5
0.25
0.25

17
OH

338

D-SER(TBU)
0.062
6.25
0.25
3.91
0.062
0.12

18
OH

339

D-THR(TBU)
0.25
25
1.56
25
0.78
0.78

20
OH

340

L-VAL
0.25
12.5
0.78
3.12
0.25
0.39

21
OH

341

L-(+)-LACTIC ACID
3.12
25
6.25
25
1.56
3.12

24
OH

342

PIPERAZIN-1-YL ACETIC ACID HYDRATE
1.56
25
1.56
6.25
0.78
0.78

25
OH

343

D-TRP
0.25
12.5
0.39
6.25
0.25
0.25

26
OH

344

L-GLU(OBZL)
1.56
>25
1.56
12.5
0.78
0.78

30
OH

345

L-HYP(TBU)
0.78
25
3.12
25
1.56
1.56

34
OH

346

4-CARBOXY METHYL- PIPERAZINE
0.78
25
3.12
6.25
0.78
1.56

36
OH

347

L-ALA-GLY
0.12
12.5
0.78
6.25
0.39
0.39

37
OH

348

L-ILE
0.25
12.5
0.78
3.12
0.39
0.39

38
OH

349

L-LEU
0.78
12.5
1.56
12.5
0.78
0.78

41
OH

350

L-MET
0.25
12.5
0.78
12.5
0.39
0.39

42
OH

351

L-NLE
0.39
12.5
0.78
6.25
0.39
0.39

56
OH

352

D-DPR(DDE)
0.12
12.5
0.78
12.5
0.25
0.25

56-3
OH

353

D-DPR(DDE) Deprotect/Rearrange
0.12
6.25
0.25
6.25
0.25
0.25

61
OH

354

D-LYS (CARBAMYL)
0.12
12.5
0.25
12.5
0.12
0.12

62
OH

355

D-ORN (CARBAMYL)
0.12
12.5
0.25
12.5
0.25
0.25

63
OH

356

D-SER
0.78
25
1.56
12.5
0.78
0.78

64
OH

357

D-THR
0.39
25
1.56
12.5
0.78
0.78

67
OH

358

GLY-GLY-GLY
0.78
12.5
1.56
6.25
0.78
0.78

68
OH

359

GLY-GLY
0.39
12.5
1.56
6.25
0.78
0.78

75
OH

360

L-LYS(BIOTIN)
0.39
25
3.12
25
1.56
1.56

79
OH

361

L-LYS(FOR)
0.78
25
3.12
26
1.56
1.56

81
OH

362

L-MET(O)
0.78
25
1.56
25
0.78
1.56

82
OH

363

L-MET(O2)
0.39
25
1.56
12.5
0.78
0.78

83
OH

364

L-ORN(PYRAZINYL CARBONYL)
0.25
25
0.78
12.5
0.39
0.78

88
OH

365

L-SER
0.78
12.5
1.56
25
0.78
1.56

89
OH

366

L-THR
0.78
>25
3.12
12.5
1.56
1.56

91
OH

367

N-ALPHA-L- GLUTAMINE
0.78
25
1.56
12.5
0.78
0.78

100
OH

368

2-(2-METHOXY ETHOXY)ACETIC ACID
0.78
6.25
1.56
6.25
0.25
0.78

108
OH

369

4-NITROBENZOYL- GLYCYL-GLYCINE
1.56
>25
3.12
>25
0.78
0.78

135
OH

370

BOC-D-ASN
0.39
>25
3.12
12.5
1.56
1.56

136
OH

371

BOC-D-GLN
0.78
>25
1.56
12.5
0.78
0.78

188
OH

372

N-ALPHA- CARBOETHOXY-L- ASPARAGINE
0.062
6.25
0.39
3.12
0.39
0.39

257
OH

373

3-AMINOBENZOIC ACID
0.39
25
1.56
12.5
0.78
0.78

287
OH

374

ETHOXYACETIC ACID
1.56
25
3.12
12.5
1.56
3.12

303
OH

375

METHOXYACETIC ACID
1.56
25
3.12
12.5
1.56
1.56

328
OH

376

D-ME-VAL
0.12
12.5
0.39
12.5
0.25
0.25

329
OH

377

D-ME-LEU
0.031
6.25
0.12
6.25
0.25
0.12

330
OH

378

L-ME-ILE
0.12
12.5
0.78
3.12
0.25
0.25

331
OH

379

L-ME-SER(BZL)
0.39
12.5
1.56
6.25
0.78
0.78

332
OH

380

L-CYS(BZL)
1.56
25
3.12
12.5
1.56
1.56

333
OH

381

L-CYS(TBU)
1.56
25
3.12
25
1.56
1.56

334
OH

382

D-CYS(TBU)
0.12
25
0.78
12.5
0.25
0.25

335
OH

383

D-ILE
0.12
12.5
0.39
12.5
0.12
0.25

336
OH

384

D-LEU
0.12
6.25
0.25
12.5
0.25
0.25

337
OH

385

D-NVA
0.062
12.5
0.25
12.5
0.12
0.12

338
OH

386

D-SER(BZL)
0.25
12.5
1.56
25
1.56
0.39

339
OH

387

D-VAL
0.12
12.5
0.25
12.5
0.25
0.25

340
OH

388

L-ME-VAL
0.39
12.5
1.56
25
0.39
0.39

341
OH

389

L-NVA
0.78
12.5
1.56
25
0.78
0.78

342
OH

390

L-SER(BZL)
0.39
25
1.56
12.5
0.78
0.78

343
OH

391

L-SER(TBU)
0.39
25
3.12
12.5
1.56
0.78

344
OH

392

L-THR(TBU)
0.78
25
3.12
12.5
1.56
1.56

345
OH

393

L-CYS(STBU)
0.39
25
3.12
12.5
0.78
0.78

517
OH

394

TS5017 (Cl-Biphenyl Vancomycin)
0.062
12.5
0.25
12.5
0.25
0.25

518
OH
X—H
TS5018 (Des-Lecyl
1.56
12.5
3.12
25
3.12
6.25

Cl-Biphenyl Vanc)

519
I

395

6′-Deoxy-6′-Iodo
0.78
>25
3.12
>25
1.56
3.12

520
I

396

6′-Deoxy-6′-Iodo- ChloroBiphenyl
0.12
12.5
0.78
12.5
0.78
0.78

521
NH2

397

6′-Deoxy-6′-Amino TS1017
0.12
12.5
0.78
6.25
0.78
0.78

522
OH
X—H
Des-Lecyl
>25
>25
>25
>25
>25
>25

Vancomycin

523
NH2

398

6′-Deoxy-6′-Amino TS5017
0.12
6.25
0.25
6.25
0.78
0.39

524
OH

399

D-SER(ET)
0.25
25
1.56
12.5
0.39
0.39

525
OH

400

D-SER(ME)
0.25
25
1.56
25
0.39
0.39

526
OH

401

N-Methy-L- SER(TBU)
0.25
25
0.78
6.25
0.39
0.39

527
OH

402

N-Methyl-D-SER(ET)
0.25
25
1.56
12.5
0.39
0.39

528
OH

403

D-SER(ISOPROPYL)
0.12
25
0.78
12.5
0.25
0.25

529
OH

404

L-SER(ISOPROPYL)
0.12
25
0.78
12.5
0.25
0.25

554
OH

405

L-HIS
0.39
12.5
3.12
25
1.56
0.78

555
OH

406

D-HIS
0.39
12.5
1.56
12.5
0.78
0.39

556
OH

407

D-GLY-(2-Pyridyl)
0.39
25
0.78
25
0.78
0.39

557
OH

408

L-Phe(3-Nitro-2- Hydroxy)
0.39
25
1.56
25
1.56
1.56

[0325]

	Number	Date	Country
Parent	09540761	Mar 2000	US
Child	10361603	Feb 2003	US

Desleucyl glycopeptide antibiotics and methods of making same

Information

Publication Number

Date Filed

Date Published

Inventors

CPC

US Classifications

International Classifications

Abstract

Description

Claims

Provisional Applications (1)

Continuations (1)