EP1147129A1 - Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface - Google Patents

Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface

Info

Publication number
EP1147129A1
EP1147129A1 EP00904285A EP00904285A EP1147129A1 EP 1147129 A1 EP1147129 A1 EP 1147129A1 EP 00904285 A EP00904285 A EP 00904285A EP 00904285 A EP00904285 A EP 00904285A EP 1147129 A1 EP1147129 A1 EP 1147129A1
Authority
EP
European Patent Office
Prior art keywords
compound
group
heterocyclic
alkyl
hydrogen
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00904285A
Other languages
German (de)
English (en)
Other versions
EP1147129A4 (fr
Inventor
Daniel Kahne
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Princeton University
Original Assignee
Princeton University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Princeton University filed Critical Princeton University
Publication of EP1147129A1 publication Critical patent/EP1147129A1/fr
Publication of EP1147129A4 publication Critical patent/EP1147129A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
    • C07K9/006—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure
    • C07K9/008—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure directly attached to a hetero atom of the saccharide radical, e.g. actaplanin, avoparcin, ristomycin, vancomycin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to saccharide compounds having transglycosylase inhibitory activity linked to non-saccharide compounds that bind to molecules located at the bacterial cell surface (e.g., cell-surface peptides or proteins).
  • the compounds of this invention are useful as antibacterial agents.
  • Peptidoglycan synthesis in bacteria is known to proceed in stages, the last of which involves transglycosylation of the disaccharide building blocks and cross-linking of the peptide chains attached thereto.
  • Compounds that inhibit any stage of peptidoglycan synthesis are potentially useful as antibacterial agents.
  • the only recognized commercialized class of compounds that inhibit the transglycosylation step comprises moenomycin and its derivatives.
  • a disaccharide fragment of moenomycin inhibits transglycosylase activity. This disaccharide, as shown below,
  • the glycopeptide antibacterial agents are believed to inhibit peptidoglycan synthesis by functioning as peptide binders. These compounds bind D-alanyl-D-alanine, preventing transpeptidation by sequestering the peptide substrates.
  • 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.
  • 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.
  • the aromatic side-chains of amino acids 2, 4, and 6 are fused together through ether linkages.
  • the side-chains of amino acids 5 and 7 are joined via a carbon-carbon bond.
  • Amino acids 1 and 3 are leucine and asparagine, respectively.
  • glycopeptide antibacterial agents are similar to vancomycin in that they have 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 vancomycin and some other glycopeptides is linked through its vicinal hydroxyl position to the amino sugar, L-vancosamine.
  • the sugars have been separately removed from vancomycin, and it has been found that the presence of both sugars enhances the activity of this class of antibacterial agents.
  • glycopeptide antibacterial agents In addition to the glycopeptide antibacterial agents, other compounds are known that bind to molecules located at the cell surface.
  • Compounds that bind to Lipid II include nisin, mersacidin, actagardine, and other antibiotics, as well as ramoplanin.
  • Compounds that bind to proteins located at the cell surface include the beta lactams and related antibiotics such as cephalosporins, carbapenems, and imipenems.
  • Other compounds thought to bind to molecules located at the bacterial cell surface include daptamycin and bacitracin. There has been no suggestion that linking these molecules or portions thereof to saccharide compounds having transglycosylase inhibitory activity would improve antibacterial efficacy.
  • This invention is directed to a compound which comprises: (i) a saccharide compound having transglycosylase inhibitory activity; and (ii) a second compound that is capable of binding a protein or enzyme involved in cell wall biosynthesis; a precursor used in cell wall biosynthesis; and/or the cell wall surface.
  • the saccharide compound is linked, directly or through a linker (e.g., a difunctional linker), to the second compound.
  • the second compound is a non-saccharide compound, provided that: when the non-saccharide compound is a hexapeptide or a heptapeptide and the saccharide compound does not contain a phosphate or phosphonate ester, then the saccharide compound is not linked directly to the non-saccharide compound through a glycosidic linkage at A4.
  • the saccharide compound is a disaccharide comprising two hexose residues joined by an alpha glycosidic linkage, and the non- saccharide compound is a peptide.
  • this invention is directed to a glycopeptide in which a disaccharide comprising two hexose residues joined by an alpha glycosidic linkage is linked to a peptide directly through a non-glycosidic linkage, or through a difunctional linker.
  • the peptide has the formula A ⁇ -A 2 -A 3 -A 4 -A 5 -A 6 -A , in which each dash represents a covalent bond; wherein the group Ai comprises a modified or unmodified -am ino acid residue, hydrogen, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic- carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl, arylsulfonyl, guanidinyl, carbamoyl, or xanthyl; wherein each of the groups A 2 to A 7 comprises a modified or unmodified -amino acid residue, whereby (i) the group Aj is linked to an amino group on the group A 2 , (ii) each of the groups A 2 , ⁇ and A 6 bears an aromatic side chain, which aromatic side chains are cross
  • R Y 2 Y ⁇ is bonded to a ring carbon atom adjacent to the alpha glycosidic linkage;
  • Rj, R 2 and R 3 are independently hydrogen, alkyl, aryl, aralkyl, alkylsulfpnyl, arylsulfonyl, aralkylsulfonyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-alkyl, heterocyclic- carbonyl or heterocyclic-alkyl-carbonyl;
  • t, R 5 , R$ and R 7 are independently hydrogen, or a hydroxyl, amino or thiol protecting group;
  • Wi, W 2 , W 3 and W 4 are independently O, NH or S;
  • Re is hydrogen, hydroxyl or a hydroxyl protecting group;
  • k, m, n, p and r are independently 0 or 1;
  • Xi is a single bond, O, NR
  • R. is not hydrogen; when Y 2 is C(O)O, C(O)S, C(S)O, C(S)S or C(NRj 2 )O, then R 2 is not hydrogen; and when Z 2 is C(O)O, C(O)S, C(S)O, C(S)S or C(NR ⁇ 2 )O, then R 3 is not hydrogen.
  • a saccharide compound exhibiting transglycosylase inhibitory activity e.g., the vancomycin disaccharide bearing a lipid-like substituent on the vancosamine nitrogen and analogs and derivatives thereof
  • a linker to non-saccharide compounds that bind to molecules located at a bacterial cell surface (e.g., the vancomycin aglycone).
  • the lipid-like substituent comprises a biaryl moiety in which the aromatic groups are joined directly or via a substituted or unsubstituted linker comprising one or more atoms, including heteroatoms.
  • One preferred biaryl moiety is a chlorophenylbenzyl group.
  • covalent attachment of the disaccharide is accomplished directly or via an amine (or diamine) linker to a carboxyl group of an aglycone.
  • "unnatural" aglycones are utlized, such as certain peptide-binding dyes, e.g., rhodamine and the like.
  • a disaccharide coupled to a peptide- binding dye, e.g., rhodamine, via a linker provides rhodamine conjugates 5a and 5b. It is contemplated that such rhodamine conjugates could target peptides found, for example, in the developing bacterial cell wall. In doing so, the rhodamine conjugate would bring the inhibitory disaccharide moiety closer to enzymes involved in cell wall biosynthesis.
  • This invention is also directed to a chemical library comprising a plurality of these compounds.
  • the invention is further directed to a method for preparing compounds in which a functionalized dalbaheptide aglycone is linked to a disaccharide transglycosylase inhibitor.
  • Figure 1 is a graph showing the effects on macromolecular synthesis in Bacillus megaterium MB410 of known antibacterial agents.
  • Figure 2 is a graph showing the effect of compound 6a on synthesis of RNA, DNA, protein and peptidoglycan in comparison with the effects of vancomycin and ampicillin.
  • Figures 3 A and 3B are graphs showing the activity of compound 6a in ether-treated bacteria, and the site of inhibition of peptidoglycan synthesis.
  • Figure 4 is a table presenting results obtained on selected compounds disclosed herein for synthesis of RNA, DNA, protein and peptidoglycan, and for the site of inhibition of peptidoglycan synthesis in ether treated bacteria.
  • Figure 5 is a table presenting results obtained on selected compounds disclosed herein for synthesis of RNA, DNA, protein and peptidoglycan, and for the site of inhibition of peptidoglycan synthesis in ether treated bacteria.
  • Figure 6 is a table presenting results obtained on selected compounds disclosed herein for synthesis of RNA, DNA, protein and peptidoglycan, and for the site of inhibition of peptidoglycan synthesis in ether treated bacteria.
  • Figure 7 is a table presenting results obtained on selected known substances of RNA, DNA, protein and peptidoglycan, and for the site of inhibition of peptidoglycan synthesis in ether treated bacteria.
  • Figure 8 is a table presenting the MIC values of selected conjugates of the invention.
  • Figure 9 exhibits a preferred synthetic scheme for the preparation of rhodamine linked disaccharides (or rhodamine conjugates) of the invention.
  • Figure 10 is a table presenting the MIC values of selected rhodamine conjugates of the invention.
  • alkyl refers to an acyclic or non-aromatic cyclic group having from one to twenty carbon atoms connected by single or multiple bonds.
  • An alkyl group may be substituted by one or more of halo, hydroxyl, protected hydroxyl, amino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, COOH, aroyloxy, alkylamino, dialkylamino, trialkylarnmonium, alkylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, heterocyclic, CONH 2 , CONH-alkyl, CONH-aryl, CONH-aralkyl, CON(alkyl) 2 , COO-aralkyl, COO-aryl, COO-heterocyclic, COO-alkyl or phosphonium
  • 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, hydrazino, alkylhydrazino, arylhydrazino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, aroyloxy, alkylamino, dialkylamino, trialkylarnmonium, alkylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, COO-alkyl, COO-aralky
  • 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 of alkyl, aryl, aralkyl, halo, hydroxyl, protected hydroxyl, amino, hydrazino, alkylhydrazino, arylhydrazino, nitro, cyano, alkoxy, aryloxy, aralkyloxy, aroyloxy, alkylamino, dialkylamino, trialkylammonium, alkylthio, alkanoyl, alkanoyloxy, alkanoylamido, alkylsulfonyl, arylsulfonyl, aroyl, aralkanoyl, COO-alkyl, COO-aralkyl, COO-aryl, COO-heterocyclic, CONH 2 , CONH-alkyl, CONH-aryl, CONH-aralkyl, CON(alkyl) 2 or phosphonium substituted by any combination of alkyl, aryl
  • alkoxy refers to groups derived from bonding an oxygen atom to an alkyl, aryl or aralkyl group, respectively.
  • alkanoyl refers to groups derived from bonding a carbonyl to an alkyl, aryl or aralkyl group, respectively.
  • heterocyclic-alkyl and “heterocyclic-carbonyl” refer to groups derived from bonding a heterocyclic group to an alkyl or a carbonyl group, respectively.
  • heterocyclic-alkyl-carbonyl refers to a group derived from bonding a heterocyclic-alkyl group to a carbonyl group.
  • hydroxyl protecting group refers to a group bonded to a hydroxyl group which is easily removed to regenerate the free hydroxyl group by treatment with acid or base, by reduction, or by exposure to light.
  • Exemplary hydroxyl protecting groups include, without limitation, acetyl, chloroacetyl, pivaloyl, benzyl, benzoyl, p-nitrobenzoyl, tert-butyl-diphenylsilyl, allyloxycarbonyl and allyl.
  • amino protecting group and thiol protecting group refer to groups bonded to an amino or thiol group, respectively, which are easily removed to regenerate the free amino or thiol group, respectively, by treatment with acid or base, by reduction, or by exposure to light.
  • Exemplary amino protecting groups include, without limitation, Fmoc, CBz, aloe and alkanoyl and alkoxycarbonyl groups.
  • Exemplary thiol protecting groups include, without limitation, alkanoyl and aroyl groups.
  • a “difunctional linker” is a group with two points of attachment, one of which is attached to the non-saccharide compound and the other to the saccharide compound.
  • the attachment to the non-saccharide compound joins the difunctional linker to, e.g., a hydroxyl, amino or carboxyl group on the non-saccharide compound.
  • the attachment to the saccharide compound joins the difunctional linker to a hydroxyl, amino or carboxyl group on the saccharide compound.
  • Exemplary linkers are derived from dicarboxylic acids, including carboxyl-substituted amino acids, which form ester or amide linkages at the points of attachment, diols, which form ethers or ester linkages at the points of attachment, or from compounds of mixed functionality, e.g., hydroxy acids.
  • Other exemplary linkers are derived from reductive alkylation with a compound bearing an aldehyde group and a carboxyl or hydroxyl group, or from alkylation of a hydroxyl or amino group with an ⁇ -halocarboxylate and subsequent esterification at the carboxyl group.
  • a “glycopeptide” is a compound comprising a peptide linked to at least one carbohydrate.
  • peptide refers to a substance containing from 2 to 20 amino acid residues linked by peptide bonds.
  • protein refers to a substance containing more than 20 amino acid residues linked by peptide bonds.
  • aglycone is the result of removing the carbohydrate residues from a glycopeptide, leaving only a peptide core.
  • a “pseudoaglycone” is the result of removing only one of two sugar residues of a disaccharide residue linked to residue A A of a glycopeptide.
  • a pseudoaglycone comprises an aglycone in which A is linked to a monosaccharide residue.
  • 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 -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 antibacterial agents.
  • 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 acid 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., vancomycin and teicoplanin.
  • 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.
  • DMF N,N-dimethylformamide
  • THF tetrahydrofuran
  • THF trifluoroacetic acid
  • EtOAc ethyl acetate
  • MeOH methanol
  • MeCN acetonitrile
  • Tf ' trifluoroacetyl group
  • DMSO dimethyl sulfoxide
  • DTBMP 2,6-di-tert-butyl-4-methylpyridine
  • DIEA diisopropylethylamine
  • AH in structural formulas refers to the allyl group
  • Fmoc refers to 9-fluorenylmethyloxycarbonyl
  • HBt 1-hydroxybenzotriazole and "OBt” to the 1- oxybenzotriazolyl group
  • PyBOP refers to be
  • a compound which comprises: (i) a saccharide compound having transglycosylase inhibitory activity; and (ii) a non-saccharide compound that is capable of binding a molecule located at a bacterial cell surface, the saccharide compound being linked directly, or indirectly through a linker, to the non-saccharide compound, provided that when the non-saccharide compound is a hexapeptide or a heptapeptide and the saccharide compound does not contain a phosphate or phosphonate ester, then the saccharide compound is not covalently bound directly to the non- saccharide compound via a glycosidic linkage.
  • Non-saccharide compounds according to the invention include, but are not limited to, "natural" and unnatural" aglycones.
  • the unnatural aglycone can be selected from peptide-binding dyes.
  • compounds are provided, which comprise: (i) a saccharide compound having transglycosylase activity; and (ii) a non-saccharide compound that is capable of binding a cell-surface peptide or protein.
  • the saccharide compound is linked directly, through a glycosidic or non-glycosidic linkage, or indirectly, through a difunctional linker, to the non-saccharide compound; provided that: when the non-saccharide compound is a hexapeptide or a heptapeptide, e.g., the peptide cores of the naturally occurring glycopeptide antibacterial agents, and the saccharide compound does not contain a phosphate or phosphonate ester, then the saccharide compound is not linked directly to the non-saccharide compound through a glycosidic linkage.
  • Non-glycosidic linkages include, without limitation, ether linkages between non-anomeric saccharide hydroxyl groups and hydroxyl groups on the non-saccharide compound, and ester linkages between saccharide hydroxyl groups and carboxyls on the non-saccharide compound or between saccharide carboxyls and hydroxyl groups on the non-saccharide compound.
  • association constant For the purposes of this invention, to be considered capable of binding a cell-surface peptide or protein, a compound must have an association constant of at least 10 3 .
  • Methods for determining association constants by NMR, fluorescence, uv or CD techniques are well known in the art. See, e.g., D.H. Williams et al. (1991).
  • Suitable saccharide compounds include, without limitation, moenomycin, the moenomycin disaccharide fragment shown hereinabove, other moenomycin derivatives having transglycosylase activity, and a disaccharide comprising two hexose residues joined by an alpha glycosidic linkage.
  • the saccharide compound is a disaccharide comprising two hexose residues joined by an alpha glycosidic linkage.
  • Suitable non-saccharide compounds that bind cell-surface molecules include, without limitation, synthetic receptors capable of binding cell-surface peptides, e.g., those reported in Still (1996), and natural peptides such as the dalbaheptides that are capable of binding cell- surface peptides or proteins, including but not limited to the terminal D-alanyl-D-alanine or D-alanyl-D-lactate units of immature cell-surface peptidoglycan.
  • the non-saccharide compound is a peptide. There is no fixed upper limit on the number of amino acid residues in the peptide because only that portion of the peptide which acts as a binding site is significant in determining activity of the compounds.
  • Suitable non-saccharide compounds that bind cell-surface proteins include, without limitation, non-saccharide inhibitors of penicillin binding proteins or other enzymes displayed on the bacterial surface.
  • the cell-surface peptide comprises D- alanyl-D-alanine or D-alanyl-D-lactate, preferably as a terminal unit, i.e., the non-saccharide compound is capable of binding D-alanyl-D-alanine or D-alanyl-D-lactate.
  • this invention is directed to a glycopeptide in which a disaccharide comprising two hexose residues joined by an alpha glycosidic linkage is linked to a peptide directly through a non-glycosidic linkage, or through a difunctional linker.
  • the peptide has the formula in which each dash represents a covalent bond; wherein the group Ai comprises a modified or unmodified -am ino acid residue, hydrogen, alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic- carbonyl, heterocyclic-alkyl, heterocyclic-alkyl-carbonyl, alkylsulfonyl, arylsulfonyl, guanidinyl, carbamoyl, or xanthyl; wherein each of the groups A 2 to A 7 comprises a modified or unmodified -amino acid residue, whereby (i) the group Ai is linked to an amino group on the group A 2 , (ii) each of the groups A 2 , t and A 6 bears an aromatic side chain, which aromatic side chains are cross-linked together by two or more covalent bonds, and (iii) the group A 7 bears a terminal
  • the disaccharide comprises two hexose residues joined by an alpha glycosidic linkage. At least one of the hexose residues is substituted by a lipid group, i.e., an organic functional group having from 2-30 carbon atoms, preferably 2-20 carbon atoms and may also contain heteroatoms.
  • the lipid group may be linear, branched, or cyclic, and may include aliphatic, aromatic and/or heterocyclic groups. A number of substituents can also be present on the hexose rings, in particular the ring not bearing the lipid group.
  • the disaccharide thus substituted possesses transglycosylase inhibitory activity.
  • the disaccharide compound has the formula (II)
  • R 2 Y 2 Y ⁇ is bonded to a ring carbon atom adjacent to the alpha glycosidic linkage;
  • Rj, R 2 and R 3 are independently hydrogen, alkyl, aryl, aralkyl, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-alkyl, heterocyclic- carbonyl or heterocyclic-alkyl-carbonyl;
  • t , R 5 , R$ and R are independently hydrogen, or a hydroxyl, amino or thiol protecting group;
  • Wj, W 2 , W 3 and W are independently O, NH or
  • Xi is a single bond, O, NR 9 or S
  • X 2 is O, NR ]2 , S, C(O)O, C(O)S, C(S)O, C(S)S, C(NR 12 )O or C(O)NR 12
  • Y x is a single bond, O, NR ]0 or S
  • Y 2 is O, NR , S, C(O)O, C(O)S, C(S)O, C(S)S, C(NR, 3 )O or C(O)NR 13
  • Z x is a single bond, O, NR n or S
  • Z 2 is O, NR 14 , S, C(O)O, C(O)S, C(S)O, C(S)S, C(NR M )O or C(O)NR, 4 ;
  • 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, alkyloxycarbonyl, aralkyloxycarbonyl, aryloxycarbonyl, alkylsulfonyl, arylsulfonyl, heterocyclic, heterocyclic-alkyl or heterocyclic- carbonyl substituents.
  • Examples of preferred protecting groups include acetyl, allyloxycarbonyl (aloe), 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 A 2 , At and A 6 , may be cross- linked through their aromatic substituent groups.
  • residues A 2 to A 7 of the glycopeptide are linked sequentially by peptide bonds and are cross-linked as in a dalbaheptide, as defined hereinabove.
  • the preferred glycopeptides thus have a peptide core in which the residues are linked as in the natural glycopeptide antibacterial agents, including, e.g., vancomycin, teicoplanin, ristocetin, avoparicin and chloroeremomycin. Substitution of different amino acids at A 3 is permitted, as are modified amino acid residues at all positions, as described hereinabove.
  • residue Ai is an -amino acid, which may be substituted on the terminal amino group by alkyl, aryl, aralkyl, alkanoyl, aroyl, aralkanoyl, heterocyclic, heterocyclic-carbonyl, heterocyclic alkyl, alkylsulfonyl, arylsulfonyl, guanidinyl, carbamoyl, or xanthyl, and the structures and interconnections of Ai to A are those of vancomycin, i.e., the glycopeptide has the heptapeptide core of vancomycin, subject to the amino acid modifications and substitutions on Ai and A 7 described hereinabove.
  • R Y 2 Y ⁇ group is attached to the anomeric position of a monosaccharide and the alpha glycosidic linkage is attached to the 2-position of the same monosaccharide. It is further preferred that Wi, W 2 and W 3 are O. It is also preferred that at least one substituent on the disaccharide, not including the non-saccharide compound, is not hydroxyl, amino, protected hydroxyl or protected amino. In one embodiment of the invention, it is preferred that R 8 is hydrogen and p is 0, further preferred that k is 1 and m is 0, still further preferred that r is 1, and most preferred that Xi is a single bond and X 2 is NR ⁇ .
  • Zi is a single bond
  • Z 2 is O, S or NR ⁇ 4
  • R t , R 5 and Re are hydrogen
  • X x is a single bond
  • X 2 is NR 12
  • Yi is a single bond
  • Y 2 is O.
  • X ⁇ X 2 R ⁇ and a CH 3 group are both attached to the 3 -position of a monosaccharide.
  • the disaccharide is derived from the disaccharide component of vancomycin, which has a glucose residue attached through its 2- position to a vancosamine residue.
  • Examples of such disaccharides are shown below in Scheme 1.
  • the vancosamine residue may lack the methyl group geminal to the amine, as in compound 11.
  • Compounds 11, 6a and 6c are substituted with an N-4-(4-chlorophenyl)benzyl substituent on the vancosamine nitrogen, while compound 6b has an n-decyl substituent on the vancosamine nitrogen.
  • Compounds 11, 6a and 6b have an equatorial 2,6- dimethoxyphenyl substituent on the glucose anomeric hydroxyl, while compound 6c has an axial methoxy substituent.
  • the compounds of formula (II) are prepared by allowing a first monosaccharide having the formula
  • R 2 Y 2 Y ⁇ is bonded to a ring carbon atom adjacent to a free hydroxyl group; and none of R 2 Y 2 Y ⁇ , WiRt, W 2 R 5 and Z ⁇ Z 2 R 3 is a free hydroxyl, amino or thiol group; to react with a second monosaccharide having the formula
  • Ar is an aryl group and none of R «, R ⁇ W 3 and XjX 2 R ⁇ is a free hydroxyl, amino or thiol group; and an activating agent; via a glycosylation reaction in which an alpha glycosidic linkage is formed between the first monosaccharide and the second monosaccharide.
  • XjX 2 R ⁇ substituent after deprotection is an amino or alkylamino group, i.e., when Xi is a single bond, X 2 is NR 12 and Ri is hydrogen
  • the disaccharide is contacted with an alkylating agent capable of reacting with the amino or alkylamino group to produce an alkylated substituent.
  • suitable alkylating agents include, without limitation, alkyl halides, alkyl sulfonate esters, and aldehydes or ketones under reactive amination conditions.
  • the anomeric aryl sulfoxide group is activated by contacting it with an organic acid anhydride which will react with the sulfoxide.
  • the organic acid anhydride may be an anhydride of a sulfonic acid, of two different sulfonic acids or of a sulfonic acid and a carboxylic acid.
  • the preferred organic acid anhydride is trifluoromethanesulfonic anhydride (trifiic anhydride, Tf 2 O).
  • a non-nucleophilic mild base is also added to the reaction mixture.
  • Suitable non-nucleophilic mild bases include, but are not limited to, porphyrins, 2,6-dialkylanilines, acetamides, 2,6-dialkylpyridines and co-solvents such as ethyl acetate or ethers.
  • the preferred base is 2,6-di-tert-butyl-4-methylpyridine (DTBMP).
  • a partially protected glucose, la or lb, having one free hydroxyl group is allowed to react with a hexose bearing an anomeric sulfoxide substituent, 2, in the presence of Tf O to produce glycosylated product 3a or 3b, respectively.
  • the ⁇ -thiophenoxy substituent in 3b is converted to an ⁇ -methoxy substituent by treatment with mercury(II) trifluoroacetate and DTBMP to give 3c.
  • Treatment of 3a or 3c with hydrazine gives the partially deprotected product 4a or 4b, respectively.
  • Hydrogenation of 4a or 4b gives completely deprotected product 5a or 5b, respectively.
  • chlorobiphenyl aldehyde Reaction with 4-(4- chlorophenyl)benzaldehyde ("chlorobiphenyl aldehyde") or 1 -decanal under conditions effective for reductive animation gives products 6a-6c, as shown.
  • This approach may be used to introduce a variety of X ⁇ X 2 R ⁇ and R 2 Y 2 Y 1 substituent groups at the vancosamine nitrogen and at the glucose anomeric carbon.
  • Scheme 3 shows the reaction of a partially protected glucose la with a hexose bearing an anomeric sulfoxide substituent 7.
  • compound 7 is a desmethylvancosamine derivative.
  • the same sequence of reactions carried out in Scheme 2 produces compound 11, a desmethyl derivative of compound 6a.
  • Particular preferred compounds of this invention are those derived from the desmethyl vancomycin disaccharide and substituted on the C-6 position of the glucose residue, as well as on the vancosamine nitrogen.
  • Derivatives at the C-6 position are produced from intermediates having a mesitylenesulfonyl group at the C-6 position and a protected vancosamine nitrogen.
  • a method for functionalizing the C-6 position is described in copending application Serial No. 09/115,667, titled “Glycopeptide Antibiotics, Combinatorial Libraries of Glycopeptide Antibiotics and Methods of Producing Same," filed July 14, 1998, and which is incorporated herein by reference.
  • ⁇ Z 2 R 3 substituent groups are introduced at the glucose-6 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.
  • the iodo group is displaced by a variety of nucleophiles to produce additional C ⁇ -derivatives.
  • a preferred nucleophile is a thiol compound, especially a heterocyclic thiol.
  • Modification of an azido group at the 6-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.
  • a disaccharide is attached, directly or through a linker, to any one of several positions on the non-saccharide compound.
  • the positions of attachment include benzylic hydroxyl groups, phenolic hydroxyl groups, the terminal group on residue A , and the amino group of Ai or A 2 .
  • Ai is hydrogen and one carboxyl group of a protected dicarboxylic acid is allowed to react with the terminal amino group of A 2 .
  • the disaccharide is allowed to react through a hydroxyl or amino group to form an ester or amide linkage, respectively.
  • This linkage is glycosidic or non-glycosidic, depending on the position of the hydroxyl or amino group on the disaccharide.
  • This strategy is shown in Figure 12 of copending application, serial no. 09/115,667, titled “Glycopeptide Antibiotics, Combinatorial Libraries of Glycopeptide Antibiotics and Methods of Producing Same," filed July 14, 1998, and which is incorporated herein by reference.
  • the disaccharide is linked directly to the t phenolic hydroxyl group through a non-glycosidic linkage.
  • Disaccharides are also attached via linkers, for example, by forming an ester, amide or thioester of a carboxylic acid on a linker group attached to a peptide with a free hydroxyl, amino or thiol, respectively, on the saccharide compound.
  • Disaccharides are also attached by reaction of haloalkyl groups, or other groups containing a leaving group, with a nucleophilic group on the peptide, e.g., an unprotected amino or hydroxyl group.
  • the peptide has the structure of the vancomycin aglycone.
  • a sugar bearing a haloalkyl group is coupled to the A 4 phenolic hydroxyl group of an aglycone by displacement of halide. This method is illustrated in Example 22.
  • a sugar bearing one free amino group e.g., an aminoalkyl group
  • the chemical library of compounds of this invention is prepared to explore the effects on biological activity of varying the substituents on different parts of the molecule.
  • at least two steps are performed, each of which introduces a substituent group.
  • a combinatorial format is established in which many different predetermined substituent groups are introduced independently at each of at least two positions on the molecule, resulting in a library containing a large number of substituted compounds, wherein each possible combination of the predetermined substituent groups is represented.
  • One strategy for combinatorial synthesis to produce the chemical library of this invention comprises sequential attachment to the non-saccharide compound of two or more sugar residues bearing the desired substituent groups via glycosylation reactions.
  • the glycosylation reactions are performed with glycosyl donors bearing an activated anomeric sulfoxide group, and more preferably, they are carried out on a polymeric support.
  • a strategy for constructing a library of substituted saccharides using solid-phase glycosylation reactions is described in Liang et al. (1996).
  • a suitable resin is a cross-linked polymer insoluble in the reaction solvent which is suitably functionalized for attachment, e.g., SASRTN (Wang's resin).
  • the differentially protected hydroxyl group on the attached sugar is deprotected.
  • this hydroxyl group is freed before attachment to the resin, since the hydroxyl group does not interfere with the coupling reaction.
  • the free hydroxyl group then serves as the nucleophile in a second glycosylation reaction.
  • the hydroxyl is glycosylated, preferably in a solid phase reaction, with a variety of azido sugars.
  • the azido groups are reduced and the resulting amino groups are then derivatized.
  • the solid phase portion of the library construction can be carried out using a parallel synthesis or a mix and split strategy.
  • the carbohydrate-modified glycopeptide derivatives would then be deprotected and cleaved from the resin. This set of compounds would then be assayed for peptide binding and anti- bacterial activity.
  • Another strategy for combinatorial synthesis to produce the chemical library of this invention comprises introduction of substituent groups at various positions on the non-saccharide compound.
  • the non-saccharide compound is a peptide, A ⁇ -A -A 3 -A 4 -A 5 -A 6 -A 7
  • the preferred positions for introduction of substituent groups are the terminal amino group of Ai or A 2 , the terminal carboxyl group of A 7 and the phenolic hydroxyl groups present on A 5 and A .
  • Substituent groups may be introduced directly at these positions or indirectly, via linkers, as described hereinabove.
  • EXAMPLE 1 3-(N-benzyloxy-carbonyloxy)-4-O-acetyl-2,3,6-trideoxy-3-C-methyl- ⁇ -L- lyxo-hexopyranosyl-(l— >2)-3,4,6-tri-O-benzyl- ⁇ -glucopyranoyl 2,6-dimethoxyphenol (3a).
  • the compound la (20 mg, 0.0315 mmol) and DTBMP (32 mg, 0.158 mmol) are azeotroped with toluene 3 times and then dissolved in 2 mL Et 2 O.
  • the reaction solution is cooled to - 78 C and 0.5 mL toluene is added.
  • Triflic anhydride (6 ⁇ L, 0.0347 mmol) is added to the reaction solution, and the sulfoxide 2 (28 mg, 0.0629 mmol) in 1 mL Et 2 O is added dropwise over 10 minutes.
  • the reaction is warmed up to 0°C in 1 hour and then quenched with 3 mL of saturated aqueous NaHCO 3 solution.
  • EXAMPLE 2 Phenyl 2-(3-N-Cbz-4-O-acetyl-2,3,6-trideoxy-3-C-methyl- ⁇ -L-lyxo- hexopyranosyl)-3,4,6 ⁇ tri-O-benzyl-l -thio- ⁇ -D-glucopyranoside (3 b).
  • EXAMPLE 4 3-(N-benzyloxy-carbonyloxy)-2,3,6-trideoxy-3-C-methyl- ⁇ -L-lyxo- hexopyranosyl-(l ⁇ 2)-3,4,6- -O-benzyl- ⁇ -glucopyranoyl 2,6-dimethoxyphenol (4a).
  • EXAMPLE 6 Vancosaminyl-(l-»2)- ⁇ -glucopyranosyl 2,6-dimethoxyphenol (5a).
  • EXAMPLE 8 2-(3-N-chlorobiphenyl-vancosaminyl)- ⁇ -D-glucopyranosyl 2,6- dimethoxyphenol (6a).
  • the solution is cooled back to room temperature, concentrated and purified by reverse-phase HPLC using a PHENOMENEX LUNA Ci8 column (21.2x250 mm), 5 ⁇ m particle, eluting with a 30 min. linear gradient of 20% acetonitrile/0.1% acetic acid in water to 70% acetonitrile/0.1% acetic acid in water; flow rate of 8 mL/min. and UN detection at
  • EXAMPLE 9 2-(3-N-decyl-vancosaminyl)- ⁇ -D-glucopyranosyl 2,6-dimethoxyphenol (6b).
  • n-decyl aldehyde 4 ⁇ L, 0.0215 mmol
  • NaCNBFLt 240 ⁇ L of 1M solution in THF, 0.24 mmol
  • the reaction is monitored by analytical HPLC using a PHENOMENEX PRODIGY 5 ⁇ m ODS(3) lOOA column (250x4.6 mm), eluting a linear gradient of 0.1% TFA in water to 70%
  • reaction mixture is quenched by addition of lOOmL dimethyl sulfide, and the mixture is extracted with 20 mL saturated aqueous NaHCO 3 solution. The aqueous layer is further extracted with CH 2 CI 2 (20
  • EXAMPLE 16 Process for Introducing a Linker at the N-methyl Leucine Position.
  • Vancomycin-HCl (497 mg, 0.335 mmol) is dissolved in 4 mL water, 4 mL distilled pyridine is added, and the mixture is stirred in a 40°C oil bath. To this solution is added phenylisothiocyanate (50 mg, 0.368 mmol). After stirring for 30 minutes the organic solvents are removed from the clear solution under reduced pressure, 100 mL water is added, and the solution is frozen and lyophilized to dryness. To the resulting powder is added 4 mL of CH 2 CI 2 and 4 mL of trifluoroacetic acid. This clear solution is stirred at room temperature for 3 minutes and then evaporated under reduced pressure to dryness.
  • Vancomycin hydrochloride (317 mg, 0.213 mmol) and glycine methyl ester hydrochloride (54 mg, 0.426 mmol) is dissolved in 2 mL DMSO and 2 mL DMF and stirred at 0°C.
  • Diisopropylethylamine (186 ⁇ L, 0.3195 mmol) is added to the reaction vessel via syringe followed by HOBt HBTU (710 mL 0.45M DMF solution, 0.319 mmol). The ice bath is removed after the addition is complete.
  • EXAMPLE 18 Process for Introducing a Linker at the Glucose C-6 Position.
  • MeO-gly-Vanco-Asp-CONH-glucosamine (I) Compound E (20 mg, 0.0119 mmol) and glucosamine.HCl (8 mg, 0.0358 mmol) are premixed and azeotroped with toluene 3 times, taken in 240 ⁇ L DMF and then cooled to 0°C. Diisopropylethylamine (21 ⁇ L, 0.119 mmol) is added to reaction vessel followed by HOBt (4.8 mg, 0.0357 mmol) and pyBOP (18 mg, 0.0358 mmol). After stirring for 15 minutes, the clear solution is suspended in 45 mL acetone and stirred, centrifuged, and decanted. The solid is dried under reduced pressure and purified by reverse-phase HPLC using a
  • EXAMPLE 20 Linking a Disaccharide to A ⁇ via a Linker.
  • the clear solution is suspended in 45 mL acetone, stirred, centrifuged, and decanted.
  • the solid is dried under reduced pressure and purified by reverse- phase HPLC using a PHENOMENEX LUNA C18 column (21.2 x 250 mm), 5 ⁇ m particle size, eluting with a 30 min. linear gradient of 20% CH 3 CN/0.1% trifluoroacetic acid in water to 100% acetonitrile/0.1% trifluoroacetic acid in water; flow rate of 7 mL/min. and ultraviolet (UV) detection at 285 nm.
  • the fractions containing the product are combined and evaporated to give 36 mg (80%) product I, 62% over 2 steps.
  • PHENOMENEX LUNA Cl ⁇ column (21.2 x 250 mm), 5 ⁇ m particle, eluting with 0.1% acetic acid in water for 5 minutes and then a 30 min. linear gradient of 0.1 % acetic acid in water to 40% acetonitrile/0.1% acetic acid in water; flow rate of 7 mL/min. and ultraviolet (UV) detection at 2 ⁇ 5 nm.
  • the fractions containing the product are combined and concentrated to give K as acetic acid salt (2.6 mg, 79%).
  • Tf 2 O 200 ⁇ L, 335 mg, 1.19 mmol
  • pyridine 100 ⁇ L, 97. ⁇ mg, 1.24 mmol
  • the reaction mixture is poured into saturated NaHCO 3 solution (10 mL).
  • the organic and aqueous layers are separated, and the aqueous layer is extracted with CH 2 C1 2 (3 x 5 mL).
  • the organic layers are combined and washed with IN HC1 (10 mL) and saturated NaHCO 3 (10 mL), dried over Na 2 SO 4 , filtered, and concentrated. Purification is accomplished by flash chromatography (7%
  • the ⁇ anomer of this product is prepared in exactly the same way; spectroscopic data for this compound are identical to those given for the anomer.
  • EXAMPLE 22 Linking a Disaccharide to A t via a Haloalkyl-Substituted Saccharide.
  • Compound 6a selectively inhibited peptidoglycan synthesis and RNA synthesis.
  • the inhibition of RNA synthesis is likely not to be a secondary effect of the inhibition of peptidoglycan synthesis because ampicillin had no effect on RNA synthesis.
  • Rifampicin did not inhibit peptidoglycan synthesis.
  • Vancomycin inhibited peptidoglycan synthesis and RNA synthesis.
  • Lipid intermediate I consists of bactoprenol MurNAc- pentapeptide.
  • Lipid intermediate II consists of bactoprenol-GlcNAc-MurNAc-pentapeptide.
  • ramoplanin is an inhibitor of the transferase step in stage II.
  • the compound inhibits incorporation into all three fractions.
  • Bambermycin is the only known inhibitor of the transglycosylase step and it inhibits inco ⁇ oration into the material retained by the PVDF filters and into the fraction that is insoluble in hot SDS but not into the butanol-soluble fractions.
  • Cefoxitin inhibits transpeptidation. It only inhibits inco ⁇ oration of [ 14 C]GlcNAc into the hot SDS-insoluble fraction.
  • Compound 6a is re-tested with ETB prepared from strain W7. The selectivity test with the known antibacterial agents confirmed that inhibition of stage II steps is observable with this strain. Again, compound 6a displays a pattern of inhibition that suggests inhibition of the transglycosylase step, as shown in Figure 3B.
  • the bromide (1.7 g, 6.2 mmol) is dissolved in DMF, NaN 3 (0.8 g) is added and the mixture heated to 80 C overnight.
  • the product is purified by flash chromatography (10%-40% EtOAc/ petroleum ether) to give 1.5 g of a pale brown oil.
  • 2-O-pivaloyl-3,4,6-tri-O-benzyl glucose sulfoxide (2) (2.1 g, 3.27 mmol) and DTBMP (2 g, 9.81 mmol) are azeotroped 3x with toluene, dissolved in 40 ml EtO Ac/3 ml CH 2 C1 2 and stirred over molecular sieves for lh. The mixture is then cooled to -78 C and Tf 2 O (330 ⁇ l, 1.96 mmol) is added. The mixture is warmed to -60 C, kept there for 15 min and cooled to -78 C.
  • the phenol is dissolved in 10 ml EtO Ac/1 ml CH 2 Cl 2 and added dropwise to the activated sulfoxide. The reaction is warmed to -50 C over 1.5h and quenched with Et 2 NH.
  • the product is partially purified by flash chromatography (10%-25% EtOAc/ petroleum ether). The semi-pure product is dissolved in 6 ml water/8 ml MeOH/ 12 ml THF (two layers). LiOH (1.5 g) is added and the mixture stirred at 35 C for 25h.
  • the product is purified by flash chromatography (35% Et 2 O/ petroleum ether) to give 1.6 g.
  • the sample is filtered through celite which is then washed with 200 ml MeOH.
  • the product is then concentrated and purified by reverse-phase HPLC using a Phenomenex LUNA Cl ⁇ column (21.2 x 250 mm, 5 ⁇ m particle size), eluting with a linear gradient (15% to ⁇ 0% CH 3 CN H 2 O with 0.1%
  • the solution is purified by reverse-phase HPLC using a Phenomenex LUNA Cl ⁇ column (21.2 x 250 mm, 5 ⁇ m particle size), eluting with a linear gradient (10% to 60% CH 3 CN/H 2 O with 0.1% TFA over 60 min, retention time 43 min) to give 3 mg of the desired product.
  • ESI-MS calculated for C 90 H 99 Cl 3 N ⁇ 0 O 26 .l ⁇ 40.6 [M + H] + : l ⁇ 42
  • the aglycone (8) is prepared as described by Nagarajan, R. and Schnabel, A. A. J. Chem. Soc, Chem. Commun. (1988) 1306-1307.
  • the vancosamine sulfide is prepared similarly to Thompson, C, Ge, M. and Kahne, D. J. Am. Chem. Soc. (1999) 121: 1237-1244.
  • EXAMPLE 26 Minimum Inhibitory Concentrations for Selected Compounds.
  • MIC values selected conjugates of the present invention are presented.
  • the disaccharide moiety is linked either to the carboxyl group of the residue A 7 or to the phenyl group of the residue t .
  • the reaction is concentrated in vacuo to a tan residue.
  • the crude residue is partitioned between CH 2 C1 2 and saturated NaHCO 3 (60 mL).
  • the cloudy aqueous layer is extracted thrice with CH 2 C1 2 (60 mL).
  • the organic layers are pooled, washed with brine, dried over Na 2 SO 4 , filtered, and concentrated in vacuo to yield a yellow oil.
  • the oil is applied to a silica flash column (5 x 12 cm) and eluted with 75%, 80%) and 90% diethylether/petroleum ether.
  • Fractions containing a 3:1 mixture of glucose and mannose are concentrated in vacuo to a white solid.
  • the white solid is dissolved in toluene with heating and allowed to cool slowly to room temperature, resulting in the formation of colorless crystals of 1 (576 mg, 1.56 mmol, 28%)
  • glucose 1 200 mg, 0.54 mmol
  • 2,6-di-tert-butyl-4-methyl-pyridine 336 mg, 1.63 mmol
  • the glucose and base are azeotroped thrice from toluene and dissolved in dry CH C1 2 (2 mL) and Et 2 O (6 mL).
  • Oven dried 4A molecular sieves are added, and the clear solution stirred at -78° C for 30 minutes.
  • Triflic anhydride 0.5 mL of a 1.31 M stock solution in CH 2 C1 2 ) is then added, yielding a cloudy solution.
  • the sulfoxide flask is washed once with CH 2 C1 2 (1 mL) and added dropwise to the glucose over 5 minutes.
  • the reaction is stirred for 30 minutes at - 74° C and then allowed to warm to -15° C over 2.5 hours. Most of the starting material is consumed once the reaction reaches -60° C.
  • the reaction is quenched by adding saturated NaHCO 3 to the reaction and warming to room temperature.
  • the reaction is filtered through a cotton plug directly into a separatory funnel and extracted thrice with CH 2 C1 2 .
  • the pooled organic layers are dried over Na 2 SO 4 , filtered, and concentrated in vacuo to a light yellow oil. Purified by silica flash chromatography (2 x)
  • Disaccharide 2 (143 mg, 0.22 mmol) is dissolved in dry DMF (2 mL) in a 25 mL round bottom flask. To the clear solution is added ⁇ a ⁇ 3 (50 mg, 0.77 mmol) and KI (24 mg, 0.14 mmol). The cloudy solution is stirred at 80° C under argon for 20 hours. The reaction is concentrated in vacuo and partitioned between H 2 O/CH 2 Cl 2 (10 mL). The aqueous layer is extracted thrice with CH 2 C1 2 (10 mL). The pooled organic layers are dried over Na 2 SO 4 , filtered, and concentrated in vacuo.
  • the Amberlite is removed by filtration, the filtrate neutralized with NHUOac (200 mg) and concentrated in vacuo to an oil.
  • the oil is applied to a flash silica column (2 x 12 cm) eluting with 10% MeOH/CH 2 Cl 2 to yield the deacetylated disaccharide (3 mg, 0.0 ⁇ 0 mmol, 72%o).
  • disaccharide 3 (11 mg, 0.023 mmol).
  • the disaccharide is azeotroped thrice from toluene.
  • the disaccharide is then dissolved in dry DMF (1 mL) and chilled to 0° C under argon.
  • Rhodamine B (11 mg, 0.023 mmol), HOBT (8.4 mg, 0.053 mmol), and TBTU (17 mg, 0.053 mmol) is added, and the reaction is stirred for 1 hour at 0° C.
  • N-methylmo ⁇ holine 13 ⁇ L is then added, and the reaction is allowed to warm to room temperature.
  • the reaction is concentrated in vacuo to a pu ⁇ le residue.
  • the residue is first applied to a LH-20 sephadex gel filtration column (2 x 18 cm) and eluted with MeOH to remove some of the unreacted Rhodamine B.
  • the crude residue is applied to a silica flash column (1.5 x 14 cm) and eluted with 6.5-8% MeOH/CH 2 Cl 2 .to yield 4 as a pu ⁇ le residue (6.5 mg, 0.0074 mmol, 32%).
  • disaccharide conjugate 4 36 mg, 0.04 mmol
  • dry DMF 1.0 mL
  • glacial AcOH 1.0 mL
  • PdCl 2 (PPh3) 2 11 mg, 0.015 mmol
  • Tributyltin hydride 0.5 mL is added in 0.1 mL portions every 10 minutes and then stirred for an additional 20 minutes.
  • the reaction is diluted with H 2 O (1.0 mL) and stored overnight in the refrigerator to promote tin salt precipitation.
  • the reaction is cooled to room temperature and filtered through a 0.2 ⁇ Nylon syringe filter.
  • the MeCN is removed in vacuo from the collected fractions, the sample frozen and lyophillized to yield 5 (4 mg, 0.004 mmol,
  • EXAMPLE 28 MIC Values for Rhodamine Conjugates.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne un composé comprenant: (i) un composé saccharide présentant une activité inhibitrice de transglycosylase, et (ii) un second composé capable de se lier à une protéine ou à une enzyme impliquée en biosynthèse de paroi cellulaire, à un précurseur utilisé en biosynthèse de paroi cellulaire, à la surface de la paroi cellulaire, ou à des combinaisons de ces possibilités. Le composé saccharide est lié, directement ou au moyen d'un lieur bifonctionnel, au composé non saccharide, à condition que lorsque le composé non saccharide est un hexapeptide ou un heptapeptide et que le composé saccharide ne contient ni un ester phosphate ni un ester phosphonate, le composé saccharide ne soit pas lié directement au composé non saccharide via une liaison glycosidique. Le composé non saccharide comprend à la fois des aglycones 'naturels' (des aglycones typiquement associés à un reste hydrate de carbone) et des aglycones 'non naturels' (des substances non typiquement associées à un reste hydrate de carbone). Des aglycones non naturels peuvent être choisis, par exemple, parmi des colorants qui se lient à des peptides.
EP00904285A 1999-01-12 2000-01-12 Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface Withdrawn EP1147129A4 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US11559699P 1999-01-12 1999-01-12
US115596P 1999-01-12
PCT/US2000/000651 WO2000042067A1 (fr) 1999-01-12 2000-01-12 Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface

Publications (2)

Publication Number Publication Date
EP1147129A1 true EP1147129A1 (fr) 2001-10-24
EP1147129A4 EP1147129A4 (fr) 2003-01-15

Family

ID=22362340

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00904285A Withdrawn EP1147129A4 (fr) 1999-01-12 2000-01-12 Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface

Country Status (3)

Country Link
EP (1) EP1147129A4 (fr)
AU (1) AU2606700A (fr)
WO (1) WO2000042067A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108409827A (zh) * 2018-05-21 2018-08-17 福州大学 一种从微藻藻渣中同步提取蛋白质和多糖的方法

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6710168B1 (en) 1999-05-19 2004-03-23 The Trustees Of The University Of Princeton Glycopeptide antibiotics, combinatorial libraries of glycopeptide antibiotics and methods of producing same
WO2000004044A1 (fr) 1998-07-14 2000-01-27 Princeton University Antibiotiques glycopeptidiques, bibliotheques combinatoires d'antibiotiques glycopeptidiques, et procedes de production correspondants
AU5029700A (en) 1999-05-19 2000-12-05 Merck & Co., Inc. Glycopeptide antibacterial compounds, compositions containing same and methods of using same
WO2001057071A2 (fr) * 2000-02-04 2001-08-09 Advanced Medicine, Inc. Derives glycopeptidiques
WO2001081373A2 (fr) * 2000-04-25 2001-11-01 Merck & Co., Inc. Composes antibacteriens glycopeptidiques et procedes pour les utiliser
BRPI0516657A (pt) 2004-11-29 2008-09-16 Univ Nagoya Nat Univ Corp derivados de monÈmero antibiótico de glicopeptìdeo
TW200808818A (en) 2006-05-26 2008-02-16 Shionogi & Co Glycopeptide antibiotic derivatives
RU2481354C2 (ru) 2007-12-26 2013-05-10 Шионоги Энд Ко., Лтд. Гликозилированные гликопептидные антибиотические производные
EP3049115A1 (fr) 2013-09-23 2016-08-03 Jawaharlal Nehru Centre For Advanced Scientific Research Conjugués de vancomycine-sucre et leurs utilisations
WO2018006063A1 (fr) * 2016-07-01 2018-01-04 Cidara Therapeutics, Inc. Composés et méthodes pour le traitement d'infections bactériennes

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5206405A (en) * 1988-08-20 1993-04-27 Hoechst Aktiengesellschaft 2-alkenyloxy-3-(phosphono)oxypropanoic acids derived from the cleavage of phosphoglycolipid antibiotics
US5602009A (en) * 1988-12-23 1997-02-11 The Salk Institute For Biological Studies Dominant negative chimeras of the steroid/thyroid superfamily of receptors
AU7193494A (en) * 1993-07-21 1995-02-20 Oxford Glycosystems Ltd Saccharides, their synthesis and use
US5840684A (en) * 1994-01-28 1998-11-24 Eli Lilly And Company Glycopeptide antibiotic derivatives
EP0901629A4 (fr) * 1996-03-21 2000-02-02 Univ Princeton Bibliotheque de ligands a base d'hydrates de carbone, dosage et procede correspondants
WO1999061583A2 (fr) * 1998-05-28 1999-12-02 Incara Pharmaceuticals Corp. Composes d'echafaudage a base d'hydrate de carbone, banques combinees, et leurs procedes de construction

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
No further relevant documents disclosed *
See also references of WO0042067A1 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108409827A (zh) * 2018-05-21 2018-08-17 福州大学 一种从微藻藻渣中同步提取蛋白质和多糖的方法

Also Published As

Publication number Publication date
WO2000042067A1 (fr) 2000-07-20
AU2606700A (en) 2000-08-01
EP1147129A4 (fr) 2003-01-15

Similar Documents

Publication Publication Date Title
Nicolaou et al. Solid‐and Solution‐Phase Synthesis of Vancomycin and Vancomycin Analogues with Activity against Vancomycin‐Resistant Bacteria
FI83524B (fi) Foerfarande foer framstaellning av farmakologiskt verkande n-/n-alkanoyl-s-(2,3-dialkanoyloxi-propyl) kysteinyl/-peptider.
US7331920B2 (en) Glycopeptide antibiotics, combinational libraries of glycopeptide antibiotics and methods of producing same
US5767254A (en) Method for making amino acid glycosides and glycopeptides
KR100930982B1 (ko) 세균 감염 치료용 달바반신 조성물
WO2000059528A1 (fr) Antibiotiques glycopeptidiques des-leucyl et procede de preparation
EP1147129A1 (fr) Saccharides portes par des composes se liant a des proteines ou des peptides cellulaires de surface
KR100242682B1 (ko) 항생물질 a 40926의 아미드 유도체
CA2337923A1 (fr) Peptides antibiotiques
WO1999061583A2 (fr) Composes d'echafaudage a base d'hydrate de carbone, banques combinees, et leurs procedes de construction
N Olsufyeva et al. Synthesis, properties, and mechanism of action of new generation of polycyclic glycopeptide antibiotics
Horvat et al. Synthesis and biological activity of [Leu5] enkephalin derivatives containing d‐glucose
EP0934076A4 (fr)
US20120252741A1 (en) Novel semi-synthetic glycopeptides as antibacterial agents
Preobrazhenskaya et al. Patents on glycopeptides of the vancomycin family and their derivatives as antimicrobials: January 1999–June 2003
Yan et al. Antibiotic activities and affinities for bacterial cell wall analogue of N-demethylvancomycin and its derivatives
WO2000041710A9 (fr) Antibiotiques a glycopeptide contenant un residu desmethylvancosamine, banques combinatoires et procedes de production associes
Horvat et al. Glycoconjugates of opioid peptides-III+. A novel regioselective synthesis of 6-O-peptidyl-d-glycopyranoses using unprotected sugars
WO2000042052A1 (fr) Disaccharides a liaison alpha non substitues
KR20030011306A (ko) 지질 ⅱ의 제조 방법
US6710168B1 (en) Glycopeptide antibiotics, combinatorial libraries of glycopeptide antibiotics and methods of producing same
JP3503066B2 (ja) 合成アグルコダルバヘプチド抗生物質
CA2372392A1 (fr) Antibiotiques de glycopeptide, bibliotheques de combinaisons d'antibiotiques de glycopeptides et procedes de production de ces derniers
WO2021205372A2 (fr) Lipopeptides antibactériens, composition pharmaceutique et composition cosmétique les comprenant, et leurs utilisations
JP2005529838A (ja) リピドｉｉの製造方法

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20010809

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

AX Request for extension of the european patent

Free format text: AL;LT;LV;MK;RO;SI

A4 Supplementary search report drawn up and despatched

Effective date: 20021202

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

RIC1 Information provided on ipc code assigned before grant

Free format text: 7C 07K 7/54 A, 7C 07K 9/00 B, 7A 61K 38/00 B, 7A 61K 38/12 B, 7A 61K 38/14 B, 7A 61K 38/16 B, 7G 01N 33/53 B, 7G 01N 33/543 B, 7G 01N 33/567 B, 7G 01N 33/566 B, 7C 07K 7/58 B, 7C 07H 3/04 B

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20040803