WO2009023160A2

WO2009023160A2 - Novel inhibitors of bacterial sortase enzymes and methods of using the same

Info

Publication number: WO2009023160A2
Application number: PCT/US2008/009577
Authority: WO
Inventors: Sadanandan E. Velu; Sthanam V.L. Narayana; Aaron L. Lucias
Original assignee: The Uab Research Foundation
Priority date: 2007-08-11
Filing date: 2008-08-11
Publication date: 2009-02-19
Also published as: WO2009023160A3; WO2009023160A8

Abstract

The present disclosure describes novel small-molecule inhibitors of bacterial sortases. The identified inhibitors may be used in the methods of treatment and prevention described in the present disclosure.

Description

Novel Inhibitors of Bacterial Sortase Enzymes and Methods of Using the Same

Inventors Sadanandan E. VeIu, Sthanam V. L. Narayana and Aaron L. Lucius

The present application claims the benefit of U.S. provisional application serial no. 60/955,353, filed 8-11-2007.

FIELD OF THE DISCLOSURE The present disclosure relates generally to inhibitors of bacterial virulence and infectivity.

Specifically, the present disclosure relates to compounds having the formula I (including compounds having the formula Ia, Ib and Ic) through XII and the identification of such compounds as inhibitors of bacterial sortase enzymes. The present disclosure also relates to methods of using the compounds disclose herein, pharmaceutical compositions comprising the compounds disclosed herein and methods for producing the compounds disclosed herein.

BACKGROUND

Bacteria can cause serious or fatal infections in humans. Gram-positive bacteria capable of causing serious or fatal infections in humans include, but are not limited to, Staphylococcus, Streptococcus, Enterococcus, Bacillus, and Listeria. Infections caused by these pathogens are particularly severe and difficult to treat in immunologically compromised patients and patients with pre-existing diseases and/or conditions. These include patients suffering from infection with the Human Immunodeficiency Virus (HIV), the virus that causes AIDS, as well as patients given immune-suppressive agents for treatment of cancer or autoimmune diseases. Gram-negative bacteria can also give rise to serious or fatal infections in humans. A unique characteristic of bacterial pathogens, including Gram-positive and Gram- negative bacteria, is their surface display of proteins anchored to the cell wall. In fact, many of these molecules are known to be involved in increasing bacterial virulence which mediates the ability of bacterial cells to infect cells in a susceptible host. Thus, a possible disruption in this anchoring process of these surface proteins may prove to be an effective treatment against bacteria infection and a mechanism to decrease bacterial virulence.

Gram-positive bacteria are formidable human pathogens. For example, Staphylococci are responsible for more than a million hospital-acquired bacterial infections every year. In addition, they are to blame for a number of minor infections ranging from pustules to boils in regular day- to-day life, and for a significant number of serious infections like endocarditis, osteomyelitis, and septic arthritis (Petti, et al., Cardiol Clin 2003, 21, (2), 219-33, vii; Lobati, et al., Infection 2001, 29, (6), 333-6; Ish-Horowicz, et al., Pediatr Infect Dis J 1992, 11, (2), 82-7). Streptococcus pneumoniae is responsible for a major portion of 3 million deaths worldwide of children from pneumonia and meningitis and for a large number of deaths from pneumonia that occurs in elderly individuals. They are also responsible for a major percentage of ear infections, sinusitis, and additional severe invasive infections in asplenic persons and those with sickle cell disease. Compounding their importance is the recent steep increase in multi-drug resistance found in bacteria and the potential of 'drug resistant bacteria' to be used in biowarfare and bioterrorism (Pappas, et al., Infect Dis Clin North Am 2006, 20, (2), 395-421, x; Clarke, et al., Br J Biomed Sci 2005, 62, (1), 40-6). Hospital strains of S. aureus are usually resistant to a variety of different antibiotics. A few strains are resistant to all clinically useful antibiotics except vancomycin, and vancomycin-resistant strains are increasingly reported (Mohr, et al., Clin Infect Dis 2007, 44, (12), 1536-42; Webster, et al., Diagn Microbiol Infect Dis 2007, 57, (2), 177-81; Weigel, et al., Antimicrob Agents Chemother 2007, 51, (1), 231-8). Methicillin resistant S. aureus (MRSA) is widespread and most methicillin-resistant bacterial strains are also resistant against multiple antibiotics, necessitating the need for identifying new therapeutic agents (Baggett, et al. J Infect Dis 2004, 189, (9), 1565-73; Deresinski, S., Clin Infect Dis 2005, 40, (4), 562-73; Salerno, et al., Ann Thorac Surg 2007, 83, (5), 1888-91). Ideally, such agents would not promote the emergence of resistant strains or do so to a lesser degree than other currently used drugs.

New approaches for the prevention and treatment of bacterial infections and reduction of bacterial virulence require greater understanding of the molecular structure and mechanisms of the chosen intervention targets and of the pathogenic role played by the target in the infection process. Bacterial infections are very complex and involve the action of a large sophisticated arsenal of virulence factors, many of which are surface-bound or secreted. Surface proteins often fall into one of four functional categories: microbial adhesion to host tissues, protection from host defense mechanisms, acquisition of nutrients for bacterial growth, and secretion of toxins and invasins. These virulence factors carry out important roles in the infection process. The possibility of human interventions interrupting this process through interference with virulence is a possibility, but not yet a reality. Gram-positive bacteria such as, but not limited to, Staphylococci, are endowed with a multitude of cell-wall anchored proteins that serve as an interface between the microbe and its host. Bacterial sortases are cysteine transpeptidases that participate in secretion and anchoring of many cell wall proteins by a mechanism highly conserved in almost the entire class of Gram-positive bacteria. Sortases and sortase homologs have also been identified in certain Gram-negative bacteria and appear to use the same mechanism. Sortases, because of their control over the cellular location of multiple virulence factors, is an attractive potential target for interrupting bacterial virulence and thereby reducing the infectivity of bacteria towards a host. Surface proteins can be attached to bacterial surface in one of several fashions (Navarre, et al., Microbiol MoI Biol Rev 1999, 63, (1), 174-229; Cossart, et al., Proc Natl Acad Sci US A 2000, 97, (10), 5013-5). Proteins that are covalently attached to the cell wall share conserved regions known as the "sorting signal" or cell wall anchors (Fischetti, V. et al., MoI Microbiol 1990, 4, (9), 1603-5; 15; Schneewind, et al., Cell 1992, 70, (2), 267-81). Sortases from Gram- positive bacteria are currently grouped into four, sometimes five, families designated Sortase (Srt) A, SrtB, SrtC (Family 3), and SrtD (Families 4 and 5), with each family recognizing a slightly different sorting signal. The sortases found in gram-negative bacteria have been grouped into another separate family (Family 6). The sorting signal for SrtA includes a conserved amino acid motif, usually LPXaaTG (SEQ ID NO: 1), where Xaa can. be any one of the naturally occurring L-amino acids, while the sorting signal for SrtB includes the conserved amino acid motif NPQTXaaN (SEQ ID NO: 2), where Xaa can be any one of the naturally occurring L- amino acids. Additional sorting signals are also known to be present in Gram-positive bacteria.

In terms of the mechanism of covalent attachment of the cell surface proteins, precursor proteins are directed into a secretory pathway by their N-terminal signal peptides (Schneewind, et al., Cell 1992, 70, (2), 267-81; Navarre, et al., MoI Microbiol 1994, 14, (1), 115-21; Navarre, et al.,. J Biol Chem 1998, 273, (44), 29135-42; Perry, et al., J Biol Chem 2002, 277, (18), 16241- 8; Schneewind, et al., Science 1995, 268, (5207), 103-6; Schneewind, et al., Embo J 1993, 12, (12), 4803-11; Ton-That, et al., J Biol Chem 1997, 272, (35), 22285-92; Ton-That, et al., J Biol Chem 1998, 273, (44), 29143-9). They are translocated across the membrane and the signal peptide is cleaved (Schneewind, et al., Cell 1992, 70, (2), 267-81; Navarre, et al., MoI Microbiol 1994, 14, (1), 115-21; Navarre, et al., J Biol Chem 1998, 273, (44), 29135-42; Perry, et al., J Biol Chem 2002, 277, (18), 16241-8; Schneewind, et al., Science 1995, 268, (5207), 103-6; Schneewind, et al., Embo J 1993, 12, (12), 4803-11). Then, the C-terminal sorting signal retains the protein in the secretory pathway. The enzyme sortase acts at this point to cleave the protein at a point in the sorting signal (for example, SrtA cleaves between the threonine (T) and the glycine (G) of SEQ ID NO: 1) (Navarre, et al., MoI Microbiol 1994, 14, (1), 115-21). The end residue of the sorting signal is then covalently linked to a cell wall component of the bacteria. The bacterial cell wall target can vary depending on the bacterial species and the sortase involved. For example, in the reaction catalyzed by SrtA in Staphylococcus, the carboxyl group of the Thr is then amide-linked to the amino group of the pentaglycine cross bridges in the staphylococcal cell wall (Perry, et al, J Biol Chem 2002, 277, (18), 16241-8). This transpeptidation is also carried out by sortase enzyme (see FIG. 1).

Thus, sortase is an enzyme that recognizes a specialized cell wall "sorting signals" in proteins destined for surface attachment and covalently links them to the bacterial cell wall (Navarre, et al., Microbiol MoI Biol Rev 1999, 63, (1), 174-229). Sortase-defective strains of various pathogens were shown to be faulty in the display of surface proteins and are less virulent (Jonsson, et al., Microbes Infect 2003, 5, (9), 775-80; Mazmanian, et al., Proc Nαtl Acαd Sci US A 2000, 97, (10), 5510-5). In SrtA gene knock-out mutants of Staphylococci, surface protein precursors were found to be accumulated in the cytoplasm or to be mis-attached to the cell membrane and cell wall (mis-sorted phenotype) (Mazmanian, et al., Proc Natl Acad Sci U S A 2000, 97, (10), 5510-5). In a number of studies, individual sortase genes have been deleted and the loss-of-sortase function resulted in less virulence in several animal models of disease (S. aureus, S. gordonii, Listeria, group A streptococci, S. pneumoniae) (Jonsson, et al, Microbes Infect 2003, 5, (9), 775-80; Barnett, et al., J Bacteriol 2002, 184, (8), 2181-91 ; Bierne, et al., MoI Microbiol 2002, 43, (4), 869-81 ; Bolken, et al., Infect Immun 2001, 69, (1), 75-80; Garandeau, et al., Infect Immun 2002, 70, (3), 1382-90; Jonsson, let al., J Infect Dis 2002, 185, (10), 1417-24).

Two sortases from Staphylococcus aureus, SrtA and SrtB have been cloned, sequenced, expressed and purified (Mazmanian, et al., Science 1999, 285, (5428), 760-3; Mazmanian, et al., Proc Natl Acad Sci U S A 2002, 99, (4), 2293-8). S. aureus SrtA is the best-characterized enzyme from the family of bacterial sortases. SrtA is a 206-amino acid protein with a cysteine residue at position 184, which was shown to be critical for its function (Ton-That, et al, Proc Natl Acad Sci USA 1999, 96, (22), 12424-9). SrtB, exhibiting 23% primary sequence homology with SrtA, is the second identified S. aureus sortase. SrtA is responsible for anchoring all LPXTG- containing surface proteins including CIfA, CIfB, FnbpA, FnbpB, SdrC, and Cna. The amino acid sequence of SrtA (SEQ ID NO: 3) and SrtB (SEQ ID NO: 4) are shown in FIGS. IB and 1C, respectively. Despite an overall homology of 20-30% among different bacterial sortases, the homology in the active site regions is significantly higher.

All sortases share various degrees of homology to the archetypal sortase, SrtA of S. aureus; however, all known SrtA homologs (all sortases) contain structurally conserved active- site residues of histidine, arginine, and cysteine (SrtA residues Hisl20, Argl97, and Cysl84) (Marraffmi, et al., Microbiol MoI Biol Rev 2006, 70, (1), 192-221). This absolute conservation includes the SrtB family, which has the most divergent consensus sorting signal of all the sortase families (Maresso, et al., J Bacteriol 2006, 188, (23), 8145-52). The absolute conservation of these residues in the active-site, along with data demonstrating that the sulfhydryl moiety in the active-site is required for the transpeptidation reaction suggests that compounds interacting with or blocking access to the Cysl84 residue (or appropriate cysteine residue in the SrtA homologs) would inhibit the enzymatic activity of any SrtA homolog. Furthermore, the close proximity of the absolutely conserved Argl97 residue to the Cysl84 residue, along with data demonstrating that arginine to alanine substitutions at the 197 residue in S. aureus greatly impaired transpeptidation activity (Marraffini, et al., J Biol Chem 2004, 279, (36), 37763-70) suggests that compounds interacting with or blocking access to the Argl97 residue (or appropriate arginine residue in the SrtA homologs) would inhibit the enzymatic activity of any SrtA homolog. Sortases are virtually universal among Gram-positive bacteria. However, some examples are also found in limited number of archaeal and Gram-negative bacteria (Pallen, et al., Trends Microbiol 2001, 9, (3), 97-102). Due to their universal presence in Gram-positive bacteria and other bacterial species, sortases are attractive pharmacotherapeutic targets (Cossart, et al., Proc Natl Acad Sci U S A 2000, 97, (10), 5013-5). The sorting signal of proteins (SEQ ID NO: 1) recognized by the main sortase is nearly identical among Gram-positive bacteria (Pallen, et al., Trends Microbiol 2001, 9, (3), 97-102; Comfort, et al., Infect Immun 2004, 72, (5), 2710-22).

Altering the targeting and/ distribution of virulence factors, such as, but not limited to, surface proteins, as opposed to bacterial growth is a novel approach for reducing bacterial virulence and infectivity towards a host. As more and more pathogens become resistant to antibiotics, altering the targeting and/or distribution of cell surface proteins on the cell wall offers a novel strategy against bacterial infections. Additionally, sortase inhibitors have the potential to be powerful therapeutic agents when used in conjunction with existing traditional antibiotics as a combination therapy since these function by a complementary mechanism.

Currently there have only been a few reports of specific sortase inhibitors (Kang, et al., Biol Pharm Bull 2006, 29, (8), 1751-5; Kim, et al., Biotechnol Biochem 2003, 67, (11), 2477-9; Kim, et al., Biosci Biotechnol Biochem 2002, 66, (12), 2751-4; Scott, et al., Biochem J 2002, 366, (Pt 3), 953-8). Recently, Oh et al. identified a small molecule reversible inhibitor with a low micromolar ICs₀ value by structurally modifying a lead compound identified by random screening of a group of small molecules (Oh, et al., J Med Chem 2004, 47, (10), 2418-21). None of the existing reports have utilized the structural information of the sortase active site for the design of their inhibitors.

Therefore, there remains a need in the art to identify new inhibitors of the bacterial sortase enzymes. The crystal structures of recombinant S. aureus SrtA_Δ59 and SrtB_Δ30 were recently determined (Zong, et al., J Biol Chem 2004, 279, (30), 31383-9; Zong, et al., Structure 2004, 12, (1), 105-12). SrtA_Δ59 and SrtB_Δ30 are fully active variants of SrtA and SrtB with 59 and

30 residues shorter from the NH-terminus, respectively. The present disclosure described the identification of several new low micromolar sortase inhibitors identified through structure based inhibitor design studies. The present disclosure also described pharmaceutical compositions containing these inhibitors methods of using the identified inhibitors and pharmaceutical compositions.

BRIEF DESCRIPTION OF THE FIGURES

FIG. IA illustrates the reaction catalyzed by the S. aureus Sortase A.

FIG. IB shows the amino acid sequence of the Sortase A enzyme from S. aureus. FIG. 1C shows the amino acid sequence of the Sortase B enzyme from S. aureus.

FIG. 2 shows the structure of compound 1.

FIG. 3 shows a FlexX model of compound 1 in the SrtA^sg active site.

FIG. 4 shows the interactions of compound 1 with amino acid residues in the SrtA_Λ59 active site.

FIG. 5 shows an exemplary synthesis of compounds 1, 13, 12a and 12b. FIG. 6 shows an exemplary synthesis of compounds 14, 15, 16 and 17.

FIG. 7 shows an exemplary synthesis of compounds 18a, 18b, 19a and 19b.

FIG. 8 shows an exemplary synthesis of compound 20.

FIG. 9 shows an exemplary synthesis of compounds 21-24.

FIG. 10 shows an exemplary synthesis of compounds 25-28. FIG. 11 shows an exemplary synthesis of compounds 29-30.

FIG. 12 shows an exemplary synthesis of compounds 31-34.

FIG. 13 shows an exemplary synthesis of compounds 43-46.

FIG. 14 shows an exemplary synthesis of compounds 51-56.

FIG. 15 shows an exemplary synthesis of compound 57.

DETAILED DESCRIPTION

Definitions

As used in this specification, the followings words and phrases have the meanings as defined below, unless otherwise limited in specific instances, either individually or as part of a larger group.

As used herein, the term "alkyl", whether used alone or as part of a substituent group, includes straight hydrocarbon groups comprising from one to twenty carbon atoms. Thus the phrase includes straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like. The phrase also includes branched chain isomers of straight chain alkyl groups, including but not limited to, the following which are provided by way of example: -CH(CHj)₂, -CH(CH₃)(CH₂CH₃), -CH(CH₂CH₃)₂, -C(CH₃)₃, - C(CH₂CH₃)₃, -CH₂ CH(CH₃)₂₎ -CH₂CH(CH₃)(CH₂CH₃), -CH₂CH(CH₂CH₃);,, -CH₂C(CH₃)₃, - CH₂C(CH₂CH₃);,-, -CH(CH₃)CH(CH₃)(CH₂CH₃), -CH₂CH₂CH(CH₃)₂, CH₂CH₂CH(CH₃)(CH₂CH₃), -CH₂CH₂CH(CH₂CH₃)_Z, -CH₂CH₂C(CH₃)₃,

CH₂CH₂C(CH₂CH₃)₃, -CH(CH₃)CH₂CH(CH₃)₂, -CH(CH₃)CH(CH₃)CH(CH₃)CH(CH₃)₂, -CH(CH₂ CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃), and others. The phrase also includes cyclic alkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl and such rings substituted with straight and branched chain alkyl groups as defined above. The phrase also includes polycyclic alkyl groups such as, but not limited to, adamantyl norbornyl, and bicyclo[2.2.2]octyl and such rings substituted with straight and branched chain alkyl groups as defined above.

As used herein, the term "alkenyl", whether used alone or as part of a substituent group, includes an alkyl group having at least one double bond between any two adjacent carbon atoms. As used herein, the term "alkynyl", whether used alone or as part of a substituent group, includes an alkyl group having at least one triple bond between any two adjacent carbon atoms.

As used herein, the term "unsubstituted alkyl", "unsubstituted alkenyl", and "unsubstituted alkynyl" refers to alkyl, alkenyl and alkynyl groups that do not contain heteroatoms. The phrase "substituted alkyl", "substituted alkenyl", and "substituted alkynyl" refers to alkyl, alkenyl and alkynyl groups as defined above in which one or more bonds to a carbon(s) or hydrogen(s) are replaced by a bond to non-hydrogen or non-carbon atoms such as, but not limited to, a halogen atom, such as F, Cl, Br, and I; and oxygen atom in groups such as carbonyl, carboxyl, hydroxyl groups, alkoxy groups, aryloxy groups, and ester groups; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, enamines, imines, oximes, hydrazones, and nitriles; a silicon atom in groups such as in trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. Other alkyl groups include those in which one or more bonds to a carbon or hydrogen atom is replaced by a bond to an oxygen atom such that the substituted alkyl group contains a hydroxyl, alkoxy, aryloxy group, or heterocyclyloxy group. Still other alkyl groups include alkyl groups that have an amine, alkylamine, dialkylamine, arylamine, (alkyl)(aryl)amine, diarylamine, heterocyclylamine, (alkyl)(heterocyclyl)amine, (aryl)(heterocyclyl)amine, or diheterocyclylamine group.

As used herein, the term "unsubstituted aryl" refers to monocyclic or bicyclic aromatic hydrocarbon groups having 6 to 12 carbon atoms in the ring portion, such as, but not limited to, phenyl, naphthyl, anthracenyl, biphenyl and diphenyl groups, that do not contain heteroatoms. Although the phrase "unsubstituted aryl" includes groups containing condensed rings such as naphthalene, it does not include aryl groups that have other groups such as alkyl or halogen groups bonded to one of the ring members, as aryl groups such as tolyl are considered herein to be substituted aryl groups as described below. Unsubstituted aryl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound, however.

As used herein, the term "substituted aryl group" has the same meaning with respect to unsubstituted aryl groups that substituted alkyl groups had with respect to unsubstituted alkyl groups. However, a substituted aryl group also includes aryl groups in which one of the aromatic carbons is bonded to one of the non-carbon or non-hydrogen atoms described above and also includes aryl groups in which one or more aromatic carbons of the aryl group is bonded to a substituted and/or unsubstituted alkyl, alkenyl, or alkynyl group as defined herein. This includes bonding arrangements in which two carbon atoms of an aryl group are bonded to two atoms of an alkyl, alkenyl, or alkynyl group to define a fused ring system (e.g. dihydronaphthyl or tetrahydronaphthyl). Thus, the phrase "substituted aryl" includes, but is not limited to tolyl, and hydroxyphenyl among others. Substituted aryl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound, however.

As used herein, the term "unsubstituted aralkyl" refers to unsubstituted alkyl, alkenyl or alkyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl, alkenyl or alkyl group is replaced with a bond to an unsubstituted or substituted aryl group as defined above. For example, methyl (CH₃) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a phenyl group, such as if the carbon of the methyl were bonded to a carbon of benzene, then the compound is an unsubstituted aralkyl group (i.e., a benzyl group). Unsubstituted aralkyl groups may be bonded to one or more carbon atom(s), oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parent compound, however.

As used herein, the term "substituted aralkyl" has the same meaning with respect to unsubstituted aralkyl groups that substituted aryl groups had with respect to unsubstituted aryl groups. However, a substituted aralkyl group also includes groups in which a carbon or hydrogen bond of the alkyl, alkenyl or alkyl part of the group is replaced by a bond to a non-carbon or a non-hydrogen atom.

As used herein, the term "unsubstituted heterocyclyl" refers to both aromatic and nonaromatic ring compounds including monocyclic, bicyclic, and polycyclic ring compounds such as, but not limited to, quinuclidyl, containing 3 or more ring members of which one or more is a heteroatom such as, but not limited to, N, O, and S. Although the phrase "unsubstituted heterocyclyl" includes condensed heterocyclic rings such as benzimidazolyl, it does not include heterocyclyl groups that have other groups such as alkyl or halogen groups bonded to one of the ring members, as compounds such as 2-methylbenzimidazolyl are "substituted heterocyclyl" groups as defined below. Examples of heterocyclyl groups include, but are not limited to: unsaturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, dihydropyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl; saturated 3 to 8 membered rings containing 1 to 4 nitrogen atoms such as, but not limited to, pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl; condensed unsaturated heterocyclic groups containing 1 to 4 nitrogen atoms such as, but not limited to, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl; unsaturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, oxazolyl, isoxazolyl, oxadiazolyl; saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, but not limited to, morpholinyl; unsaturated condensed heterocyclic groups containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example, benzoxazolyl, benzoxadiazolyl, benzoxazinyl (e.g. 2H-l,4-benzoxazinyl etc.); unsaturated 3 to 8 membered rings containing 1 to 3 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, thiazolyl, isothiazolyl, thiadiazolyl (e.g. 1,2,3- thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.); saturated 3 to 8 membered rings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, thiazolodinyl; saturated and unsaturated 3 to 8 membered rings containing 1 to 2 sulfur atoms such as, but not limited to, thienyl, dihydrodithiinyl, dihydrodithionyl, tetrahydrothiophene, tetrahydrothiopyran; unsaturated condensed heterocyclic rings containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to, benzothiazolyl, benzothiadiazolyl, benzothiazinyl (e.g. 2H-l,4-benzothiazinyl, etc.), dihydrobenzothiazinyl (e.g. 2H-3,4- dihydrobenzothiazinyl, etc.), unsaturated 3 to 8 membered rings containing oxygen atoms such as, but not limited to furyl; unsaturated condensed heterocyclic rings containing 1 to 2 oxygen atoms such as benzodioxolyl (e.g. 1,3-benzodioxoyl, etc.); unsaturated 3 to 8 membered rings containing an oxygen atom and 1 to 2 sulfur atoms such as, but not limited to, dihydrooxathiinyl; saturated 3 to 8 membered rings containing 1 to 2 oxygen atoms and 1 to 2 sulfur atoms such as 1 ,4-oxathiane; unsaturated condensed rings containing 1 to 2 sulfur atoms such as benzothienyl, benzodithiinyl; and unsaturated condensed heterocyclic rings containing a sulfur atom and 1 to 2 oxygen atoms such as benzoxathiinyl. Heterocyclyl group also include those described above in which one or more S atoms in the ring is double-bonded to one or two oxygen atoms (sulfoxides and sulfones). For example, heterocyclyl groups include tetrahydrothiophene, tetrahydrothiophene oxide, and tetrahydrothiophene 1,1 -dioxide. Preferred heterocyclyl groups contain 5 or 6 ring members.

As used herein, the term "substituted heterocyclyl" refers to an unsubstituted heterocyclyl group as defined above in which one of the ring members is bonded to a non-hydrogen atom such as described above with respect to substituted alkyl groups and substituted aryl groups. Examples, include, but are not limited to, 2-methylbenzimidazolyl, 5-methylbenzimidazolyl, 5- chlorobenzthiazolyl, 1 -methyl piperazinyl, and 2-chloropyridyl among others.

The phrase "unsubstituted heterocyclylalkyl" refers to unsubstituted alkyl, alkenyl or alkynyl groups as defined above in which a hydrogen or carbon bond of the unsubstituted alkyl, alkenyl or alkynyl group is replaced with a bond to a substituted or unsubstituted heterocyclyl group as defined above. For example, methyl (-CH₃) is an unsubstituted alkyl group. If a hydrogen atom of the methyl group is replaced by a bond to a substituted or unsubstituted heterocyclyl group, such as if the carbon of the methyl were bonded to carbon 2 of pyridine (one of the carbons bonded to the N of the pyridine) or carbons 3 or 4 of the pyridine, then the compound is an unsubstituted heterocyclylalkyl group.

The phrase "substituted heterocyclylalkyl" refers to substituted alkyl, alkenyl or alkynyl groups as defined above in which a hydrogen or carbon bond of the substituted alkyl, alkenyl or alkynyl group is replaced with a bond to a substituted or unsubstituted heterocyclyl group as defined above. However, a substituted heterocyclylalkyl group also includes groups in which a non-hydrogen atom is bonded to a heteroatom in the heterocyclyl group of the heterocyclylalkyl group such as, but not limited to, a nitrogen atom in the piperidine ring of a piperidinylalkyl group.

The phrase "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine. The term "pharmaceutically acceptable salts" is^v meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, oxalic, maleic, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galacrunoric acids and the like (see, for example, Berge, S. M., et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

The term "prodrug" is meant to include functional derivatives of the compounds disclosed which are readily convertible in vivo into the required compound. Thus, in the methods of treatment of the present disclosure, the term "administering" shall encompass the treatment of the various disease states/conditions described with the compound specifically disclosed or with a prodrug which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985. The terms "prevent", "preventing", "prevention" "suppress", "suppressing" and

"suppression" as used herein refer to administering a compound prior to the onset of clinical symptoms of a disease state/condition so as to prevent any symptom, aspect or characteristic of the disease state/condition. Such preventing and suppressing need not be absolute to be useful.

The terms "treat", "treating" and "treatment" as used herein refers to administering a compound after the onset of clinical symptoms of a disease state/condition so as to reduce or eliminate any symptom, aspect or characteristic of the disease state/condition. Such treating need not be absolute to be useful.

The term "in need of treatment" as used herein refers to a judgment made by a caregiver that a patient requires or will benefit from treatment. This judgment is made based on a variety of factors that are in the realm of a caregiver's expertise, and may include the knowledge that the patient is ill as the result of a disease state/condition that is treatable by a compound or pharmaceutical composition of the disclosure.

The term "in need of prevention" as used herein refers to a judgment made by a caregiver that a patient requires or will benefit from prevention. This judgment is made based on a variety of factors that are in the realm of a caregiver's expertise, and may include the knowledge that the patient may become ill as the result of a disease state/condition that is treatable by a compound or pharmaceutical composition of the disclosure.

The term "individual" or "patient" as used herein refers to any animal, including mammals, such as, but not limited to, mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, or humans. The term may specify male or female or both, or exclude male or female; and

The term "therapeutically effective amount", in reference to the treating, preventing or suppressing of a disease state/condition, refers to an amount of a compound either alone or as contained in a pharmaceutical composition that is capable of having any detectable, positive effect on any symptom, aspect, or characteristics of the disease state/condition. In one embodiment, a therapeutically effective amount is a tumor growth inhibiting amount. Such effect need not be absolute to be beneficial. Summary of the Disclosure The present disclosure provides compounds of the general formula I (including compounds having the formula Ia, Ib and Ic) through XII, specifically compounds 1-8, 12-34 and 43-67. In one embodiment, these compounds are useful as inhibitors of bacterial sortase enzymes. In a particular embodiment, these compounds are useful as inhibitors of bacterial SrtA, SrtB, SrtC and/or SrtD, or homologs thereof. In a further particular embodiment, the compounds are useful as inhibitors of bacterial SrtA. The compounds of the present disclosure have utility in treating and/or preventing a variety of disease states and/or conditions involving bacterial infection and virulence, wherein said bacteria causing or contributing to the bacteria infection and virulence express a sortase enzyme.

In an alternate embodiment, the present disclosure provides a method of treating a disease state and/or condition in a subject which is caused by, at least in part, bacterial infection and virulence, the method comprising the administration to said subject of a therapeutically effective amount of a compound of the present disclosure or a pharmaceutical composition containing a compound of the present disclosure. In a further alternate embodiment, the present disclosure provides a method of preventing or suppressing a disease state and/or condition in a subject which is caused by, at least in part, bacterial infection and virulence, the method comprising the administration to said subject of a therapeutically effective amount of a compound of the present disclosure or a pharmaceutical composition containing a compound of the present disclosure.

In yet a further alternate embodiment, the present disclosure provides for a method of reducing or eliminating bacterial virulence in a subject, the method comprising the administration to said subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutical composition containing a compound of the present disclosure. In still a further embodiment, the present disclosure provides for methods of treating bacterial infection in a subject, the method comprising the administration to said subject of a therapeutically effective amount of a compound of the present disclosure or a pharmaceutical composition containing a compound of the present disclosure.

In still a further embodiment, the present disclosure provides for methods of preventing or suppressing bacterial infection in a subject, the method comprising the administration to said subject of a therapeutically effective amount of a compound of the present disclosure or a pharmaceutical composition containing a compound of the present disclosure.

Other embodiments and aspects of the invention will be apparent according to the description provided below. Sortase Inhibitors

This present disclosure provides compounds of the general formula I (including compounds having the formula Ia, Ib and Ic) through XII and pharmaceutical compositions comprising the same or pharmaceutically acceptable salts thereof, or esters thereof, or prodrugs thereof and tautomers and polymorphic variants of any of the foregoing. In one embodiment, the compounds disclosed are useful as inhibitors of bacterial sortase enzymes.

With regard to compounds having the general formula (I), the structure below is provided:

(D

Where

Ring Z is selected from substituted or unsubstituted aryl or substituted or unsubstituted heterocycle; Ring Y is selected from substituted or unsubstituted aryl or substituted or unsubstituted heterocycle;

Ring X is optional and when present is selected from substituted or unsubstituted aryl or substituted or unsubstituted heterocycle;

B is selected from S or O or NRi, where Ri is H or a substituted or unsubstituted alkyl; Li and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and

L in Li and L₂ is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl; and

The linking group I, as indicated, is a C₂ to C₅ substituted or unsubstituted alkyl, C₂ to C₅ substituted or unsubstituted alkenyl containing a carbon-carbon double bond (in either the cis or trans configuration) or C₂ to C₅ substituted or unsubstituted alkynyl containing a carbon-carbon triple bond; in one embodiment, the linking group I is unsubstituted; in an alternate embodiment, the linking group I is a C₂ unsubstituted alkenyl with the carbon-carbon double bond in the trans configuration;

In a specific embodiment, ring Z is a substituted or unsubstituted 5 membered ring, including but not limited to, heterocylic compounds containing oxygen (such as furan), sulfur

(such as thiophene) or nitrogen (such as pyrrole) or a substituted or unsubstituted 6 member ring, including but not limited to, phenyl, heterocylic compounds containing oxygen (such as pyran), sulfur (such as thiapyrane) or nitrogen (such as pyridine) phenyl, or selected from substituted or unsubstituted phenyl, benzothiophene and indole and ring X is morpholine. In a specific embodiment of compounds having the general formula (I), compounds of the structure Ia are provided:

(Ia)

Where

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NRj, where R₁ is H or a substituted or unsubstituted alkyl;

E is selected from the group consisting of: CH₂, O and N;

Li and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and L in Li and L₂ is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl; The linking group I, as indicated, is a C₂ to C₅ substituted or unsubstituted alkyl, C₂ to C₅ substituted or unsubstituted alkenyl containing a carbon-carbon double bond (in either the cis or trans configuration) or C₂ to C₅ substituted or unsubstituted alkynyl containing a carbon-carbon triple bond; in one embodiment, the linking group I is unsubstituted; in an alternate embodiment, the linking group I is a C₂ unsubstituted alkenyl with the carbon-carbon double bond in the trans configuration;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORn, COORi₄, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl.

R₆ through R₈ are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl.

Ri₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₅Ri₆; Rn is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl. Ri₅ and Ri₆ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl.

In a specific embodiment, R₂ through R₅ are each independently selected from the group consisting of: H, COR_J3, COOR_H and substituted or unsubstituted alkyl. In a further specific embodiment, R₂ through R₅ are each independently selected from the group consisting of: COOCH₃, COOH, CHO, CH₂OH and CONH₂.

In a specific embodiment, R₂ is selected from the group consisting of: H, COR1₃,

COOR_H and substituted or unsubstituted alkyl while R₃ through R₈ are H, I is a C₂ alkenyl, B is

NH and 1 and 2 = 0. In a further specific embodiment, R₂ is selected from the group consisting of: COOCH₃, COOH, CHO, CH₂OH and CONH₂, while R₃ through R₈ are H, I is a C₂ alkenyl, B is NH and 1 and 2 = O.

In certain specific embodiment of the structure of compound Ia, the structures below are provided:

A is S, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 1.

A is S, B is NH, E is O, R₂ is COOCH₃, R₃ through R₈ are H, Lj and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 12a.

A is O, B is NH, E is O, R₂ is COOCH₃, R₃ through R₈ are H, L₁ and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 12b.

A is O, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 13.

A is S, B is NH, E is O, R₂ is CH₂OH, R₃ through R₈ are H, L₁ and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 18a.

A is O, B is NH, E is O, R₂ is CH₂OH, R₃ through R₈ are H, L₁ and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 18b.

A is S, B is NH, E is O, R₂ is CHO, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 19a.

A is O, B is NH, E is O, R₂ is CHO, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 19b.

A is S, B is NH, E is O, R₂ is CONH₂, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 20.

A is S, B is NH, E is O, R₂ is COOCH₃, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkynyl and the compound has the structure of compound 21.

A is S, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₂ alkynyl and the compound has the structure of compound 22.

A is S, B is NH, E is O, R₂ is COOCH₃, R₃ through R₈ are H, L₁ and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the cis configuration and the compound has the structure of compound 23.

A is S, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, L₁ and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the cis configuration and the compound has the structure of compound 24.

A is S, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, Lj and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 29.

A is S, B is NH, E is CH₂, R₂ is COOCH₃, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₃ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 31.

A is O, B is NH, E is CH₂, R₂ is COOCH₃, R₃ through R₈ are H, Lj and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 32.

A is S, B is NH, E is CH₂, R₂ is COOH, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 33.

A is S, B is NH, E is O, R₂ is COOCH₂CH₃, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 43.

A is O, B is NH, E is O, R₂ is COOCH₂-C₆Hi₂, R₃ through R₈ are H, Lj and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 44.

A is O, B is NH, E is O, R₂ is COOCH₂CH₂CH₃, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 45.

A is O, B is NH, E is O, R₂ is COOCH₂CH₂CH₂CH₃, R₃ through R₈ are H, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 46.

A is S, B is NH, E is O, R₂ is COOCH₃, R₃ through R₈ are H, Li is not present (i.e., 1 is equal to O) and L₂ is CH₃, the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 47.

A is S, B is NH, E is O, R₂ is COOH, R₃ through R₈ are H, Li is not present (i.e., 1 is equal to O) and L₂ is CH₃, the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 48.

In another specific embodiment of compounds having the general formula (I), compounds of the structure Ib are provided:

(Ib)

Where

A is selected from the group consisting of: S, CH₂, O and N; B is selected from the group consisting of: S, O and NRi, where Ri is H or a substituted or unsubstituted alkyl;

E is a heteroatom selected from the group consisting of: CH₂, O and N;

F is a single, optional N heteroatom; when present, F may be in a para, meta or ortho position with respect to the point of attachment; Li and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and

L in Li and L₂ is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORn, COOR₁₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

R₉ through R)₃ are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl; provided that when F is present at a given position (i.e., para, meta or ortho position), one of the R₉ through Ri₃ substituent groups will be absent;

R₁₄ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR₁₆R₁₇;

R₁₅ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl;

Rj₆ and Ri₇ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl.

In a specific embodiment, R₂ through R₅ are each independently selected from the group consisting of: H, CORi₄, COOR₁₅ and substituted or unsubstituted alkyl. In a further specific embodiment, R₂ through R₈ are each independently selected from the group consisting of:

COOCH₃, COOH, CHO, CH₂OH and CONH₂.

In a specific embodiment, R₂ is selected from the group consisting of: H, CORi₄,

COORi₅ and substituted or unsubstituted alkyl, while R₃ through R₅ and R₉ through Ri₃ are H, I is a C₂ alkenyl, B is NH and 1 and 2 = O. In a further specific embodiment, R₂ is selected from the group consisting of: COOCH₃, COOH, CHO, CH₂OH and CONH₂, while R₃ through R₅ and

R₉ through R_i3 are H, I is a C₂ alkenyl, B is NH and 1 and 2 = O.

In certain specific embodiment of the structure of compound Ib, the structures below are provided (see Table 2): F is N and is in the para position, B is NH, E is O, R₂ is COOCH₃, R₃ through R₅ and R₉,

Rio, Ri₂ and Ri₃ are H, Rn is absent, Lj and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 51.

F absent, B is NH, E is O, R₂ is COOCH₃, R₃ through R₅ and R₉ through R₁₃ are H, Li and L₂ are not present (i.e., 1 and 2 each equal 0), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 52.

F is N and is in the meta position, B is NH, E is O, R₂ is COOCH₃, R₃ through R₅ and R₉, Rn, Ri₂ and Rj₃ are H, R₁₀ is absent, L₁ and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 55.

F is N and is in the meta position, B is NH, E is O, R₂ is COOH, R₃ through R₅ and R₉, Rn, Ri₂ and Rj₃ are H, Ri₀ is absent, Li and L₂ are not present (i.e., 1 and 2 each equal O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 56.

In another specific embodiment of compounds having the general formula (I), compounds of the structure Ic are provided:

(Ic)

Where

A is selected from the group consisting of: S, CH₂, O and N;

L₁ is an optional linking groups, where 1 is selected from 0 or 1 and L in L₁ is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl;

The linking group I, as indicated, is a C₂ to C₅ substituted or unsubstituted alkyl, C₂ to C₅ substituted or unsubstituted alkenyl containing a carbon-carbon double bond (in either the cis or trans configuration) or C₂ to C₅ substituted or unsubstituted alkynyl containing a carbon-carbon triple bond; in one embodiment, the linking group I is unsubstituted; in an alternate embodiment, the linking group I is a C₂ unsubstituted alkenyl with the carbon-carbon double bond in the trans configuration; R₂ through R₆ are each independently selected from the group consisting of: H, OH, CORi₀,

COORi _l, NR]₄R]₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, C₁ to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl; R₇ through R₉ are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl;

Ri₀ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR]₂R]₃;

Rn is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl;

Ri₂ and R]₃ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl; and Ri₄ and R₁₅ are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl and substituted or unsubstituted aralkyl.

In a specific embodiment, R₂ through R₆ are each independently selected from the group consisting of: H, OH, CORio, COORn, NR_J4R_{I 5} and substituted or unsubstituted alkyl. In a further specific embodiment, R₂ through R₆ are each independently selected from the group consisting of: NHCH₂-Phe, NH-Phe, N(CH₂-Phe)₂, N(CH₂CH₂OH)₂, COOCH₃, COOH, CHO, CH₂OH and CONH₂.

In a specific embodiment, R₂ and R₃ are selected from the group consisting of: H, CORi₀, COORi i, NR₁₄Ri₅ and substituted or unsubstituted alkyl, while R₃ through R₉ are H, I is a C₂ alkenyl, B is NH and 1 and 2 = 0. In a further specific embodiment, R₂ and R₃ are selected from the group consisting of: NH-CH₂-Phe, NH-Phe, N-(CH₂-Phe)₂, N-(CH₂CH₂OH)₂, COOCH₃, COOH, CHO, CH₂OH and CONH₂, while R₄ through R₉ are H, I is a C₂ alkenyl, B is NH and 1 and 2 = 0.

In certain specific embodiment of the structure of compound Ib, the structures below are provided (see Table 2):

A is S, B is NH, R₂ is NH-CH₂-Ph, R₃ is COOCH₃, R₄ through R₉ are H, Li is not present (i.e., 1 equals O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 58.

A is S, B is NH, R₂ is NH-Ph, R₃ is COOCH₃, R₄ through R₉ are H, Li is not present (i.e., 1 equals O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 60.

A is S, B is NH, R₂ is N-(CH₂-Ph)₂, R₃ is COOCH₃, R₄ through R₉ are H, L₁ is not present (i.e., 1 equals 0), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 61.

A is S, B is NH, R₂ is N-(CH₂-Ph)₂, R₃ is COOH, R₄ through R₉ are H, Li is not present (i.e., 1 equals O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 62.

A is S, B is NH, R₂ is N-(CH₂CH₂OH)₂, R₃ is COOH, R₄ through R₉ are H, L, is not present (i.e., 1 equals O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 63.

A is S, B is NH, R₂ is N-(CH₂-Phe)₂, R₃ through R₉ are H, Lj is not present (i.e., 1 equals O), the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 67.

With regard to compounds having the general formula II through VIII, the following structures are provided:

With regard to compounds having the general formula (XII), the structure below is provided:

Where

A is selected from the group consisting of: S, CH₂, O and NRi₈, where Ri₈ is H or a substituted or unsubstituted alkyl

R₁ through R₉ are each independently selected from the group consisting of: H, OH, NRi₆R₁₇,

CORi₂, COOR₁₃, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, C₁ to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

R₁₀ and Rn are each independently selected from the group consisting of: H substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl; R₁₂ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR₁₄Rj ₅;

Ri₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl; Ri₄ and Rn are each independently selected from the group consisting of: H, O and substituted or unsubstituted alkyl.

In a specific embodiment, Ri through R₉ are each independently selected from the group consisting of: H, NRi₆Ri₇, COR₁₂, COORi₃ and substituted or unsubstituted alkyl. In a further specific embodiment, Ri is NO₂ or NH₂. In a specific embodiment, Ri is selected from the group consisting of: H, NRi₆Ri₇,

COR)₂, COOR]₃ and substituted or unsubstituted alkyl, while R₂ through R₉ are at H, Rj₀ through

Rn are CH₃, A is O and I is a C₂ alkenyl. In a further specific embodiment, Ri is NO₂ or NH₂, while R₂ through R₉ are at H, Ri₀ through Ri 1 are CH₃, A is O and I is a C₂ alkenyl.

In certain specific embodiment of the structure of compound XII, the structures below are provided:

A is O, Ri is NO₂, R₂ through R₉ are H, the linking group I is a C₂ alkenyl with the carbon-carbon double bond in the trans configuration and the compound has the structure of compound 57.

Any of the foregoing compounds may be provided as pharmaceutically acceptable salts, esters, prodrugs, tautomers or polymorphic variants. Compound Identification

The structures of compound 1 (FIG. 2), compound 57 (Table 2) and compounds 2-8 (Table 1) were identified as inhibitor of bacterial sortase enzymes by in-silico virtual screening against the S. Aureus SrtAΔS9 enzyme. Virtual screening of commercial small molecule libraries from Maybridge and ChemBridge (-150,000 compounds) against the S. aureus SrtA_Λ59 active site were conducted using FlexX software package integrated in SYBYL 7.0. FlexX is a fast, flexible docking method that uses an incremental construction algorithm to place ligands into the active site of the receptor. FlexX predicts the geometry of the complex as well as the free energy of binding for a given protein and a ligand. The high resolution X-ray crystal structure of both the apo enzyme and its complex with the peptide substrate were available, thus giving a clear description of the substrate binding site. In this case, FlexX is an ideal tool for screening large libraries of ligands to find leads for drug design. FlexX has the feature of allowing conformational flexibility of the ligand in complementing the active site, when active site is used as a constraint, while docking. FlexX allows complete specification of the receptor active site, including oxidation states, metal ions, side chain protonation states and automatic ligand positioning, while docking. FlexX allows the ability to dock and score small combinatorial or related libraries of compounds to identify lead structural scaffolds.

In FlexX docking, the enzyme active site environment is identified by a receptor description file (RDF). A RDF for SrtA_Δ59 was created using the receptor PDB file, the "active site file" which is constituted of all residues within 6.5 A distance from the center of the active site and a "pocket file" which is composed of a few hand-picked residues that are directly interacting with the substrate peptide. 2D compound data sets from commercial sources were transformed into three dimensional molecular structures using COMPARE program. All compounds were generated in the protonated state that can be assumed under physiological conditions. The following limits were applied for the virtual screening: molecular weight >150 <500, ClogP <5. In addition, compounds bearing metal ions were omitted from the data set. All calculations were performed on Silicon Graphics Octane (2.2 GHz RlOOOO processor) computer. Docking was carried out to obtain a population of possible conformations and orientations of the inhibitors in the active site. For each inhibitor, docking runs were performed with a maximum allowed number of 10 poses. Docking position of highest scoring conformation is individually analyzed. The structures of top scoring compounds were manually examined for their drug like properties based on Lipinski's rule⁴⁵ and for synthetic versatility.

The best scoring compounds from the in-silico virtual screen were purchased and screened against the SrtA_Λ59 protein by monitoring the effect of the compounds on the steady state cleavage of a model substrate peptide using a FRET assay, from which IC₅₀ values were determined (see experimental section). Nine compounds, compound 1 (FIG. 2), compound 57 (Table 2) and compounds 2-8 (Table 1) exhibited in vitro inhibition of the catalytic activity of the SrtA_Δ59 enzyme with IC₅₀ values ranging from about 31 to 400 μM. These compounds were obtained in milligram quantities from commercial sources. The most active compounds identified from this virtual screening were compound 1 (FIG. 2) which had an IC₅₀ value of 75 μM, compound 57 (Table 2) which had an ICs₀ value of 31 μM and compound 2 (Table 1) which had an IC₅₀ value of 97 μM. The IC₅₀ values of compounds 3 - 8 are given in Table 1.

A FlexX docking model of inhibitor 1 in SrtAΛ₅9 active site showed that the inhibitor fits well in the active site with multiple contacts (Figure 3). Inhibitor 1 has a three-ring structure. The left end is a thiophene ring (ring A), the middle ring is phenyl ring (ring B) and the right end is a morpholine ring (ring C). Ring A and B are connected by an acrylamido linkage in which the double bond has trans stereochemistry. Middle ring B bears a carboxylic acid group at a position meta to the point of attachment of acrylamido linkage. Ring C is connected to the middle ring B at the ortho position to COOH group through the morpholine ring N atom.

FlexX docking model of inhibitor 1 revealed several interactions of the inhibitor within the active site (Figure 4). Morpholine ring oxygen atom has a hydrogen bonding interaction with amide backbone NH of Trpl94. The cysteine residue (Cysl84) is at 2.94A distance from the morpholine ring carbon atom next to the oxygen. If the morpholine ring is appropriately substituted with a hydrogen bond acceptor or donor the modification may result in additional hydrogen bonding interactions. The carboxylic acid on the middle phenyl ring has a direct salt bridge interaction with the guanidine side chain of Argl97. The amide NH group of the linker has a hydrogen bonding interaction with the carboxylic acid side chain of GIu 105 and the amide carbonyl oxygen of the linker has a hydrogen bonding interaction with the OH side chain of Serl lό. The thiophene ring (ring A) of inhibitor 1 is in close proximity to Glnl72 (3.2A) and Asnl 14 (3.1 A). Appropriate substitution to the thiophene ring may result in additional hydrogen bonding interactions with Glnl72 and Asnl 14. Besides, a wide range of hydrophobic residues (Val201, Phe200, Ilel99, Leul69, Ilel58, VaIl 68, VaIl 64, Leul79, Ilel l5, Leul81, Ilel82, Ilel 17, Leu97 and PhelO3) surround the middle phenyl ring B on either side. The structural features of the lead compound along with its critical interactions with active site residues and the presence of key active site amino acid residues in close proximity to the inhibitor provide us plenty of opportunities for SAR studies and lead optimization (FIG. 4).

FlexX docking model of inhibitor 57 also revealed several interactions of the inhibitor within the active site. The carbonyl group present in the molecule showed an interaction with an arginine residue (Arg 197) that is conserved in the active site of sortase family enzymes. The nitro group present in the molecule showed critical interactions with several residues in the active site (Asp 170, GIu 171, GIn 172 and GIn 178). Furthermore, the nitrogen of dimethylamino group was found to be in close proximity to Serl 16 and Glul05. Synthesis and Biological Activity

Based on these initial studies, structure-activity relationship (SAR) studies were carried out. These studies were initially conducted using compound 1 and compound 57 as model compounds.

For SAR studies a general method for the synthesis of compound 1 was developed. The synthetic procedures for synthesis of the various analogs described were modeled around this synthesis. Chemistry employed for the synthesis of compound 1 is outlined in Scheme 1 (FIG. 5).

Commercially available methyl 2-(N-morpholino)-5-nitrobenzoate (9) was reduced using hydrogen in the presence of PaVC in anhydrous ethyl acetate to afford the corresponding amino compound 10. The compound 10 was coupled with commercially available trøns-3-(thiophene- 2-yl)acrylic acid (lla) in the presence of ethyl(N,N-dimethylaminopropyl)carbodiimide (EDAC) and iVyV-dimethylaminopyridine (DMAP) in 1 ,2-dichloroethane to form the amide compound 12a. Basic hydrolysis of the ester methyl group present in compound 12a afforded the compound 1 as a white crystalline solid. The synthesized product was found to be exactly identical to the compound obtained from commercial sources according to ¹H-NMR, ¹³C-NMR, MS and enzyme inhibition data.

Derivatives of compound 1 were synthesized in order to derive preliminary structure activity relationship data. The activities of the derivatives against the activity of the SrtAΔ₅9 protein were determined using the FRET assay as described. Structures and IC₅₀ values of the newly synthesized derivative compounds are given in Table 2.

Compound 13 is a furan analog of compound 1. Design of this structure was inspired by the comparison of activities of thiophene and furan compounds 3 and 6 (Table 1). These two compounds have the same structure except that compound 3 contained a furan ring and compound 6 contained a thiophene ring. Furan compound 3 is more than two fold more active than the thiophene compound, 6. Therefore, a furan substitution of the thiophene ring of compound 1 may increase its activity. Compound 13 was synthesized by a similar procedure as described for the synthesis of compound 1 (Scheme 1, FIG. 5). However, in the synthesis of compound 13, a trøM_?-3-(furan-2-yl) acrylic acid (lib) instead of trøns-3-(thiophene-2-yl) acrylic acid (Ha) was used. Compound 10 was coupled with commercially available trans-3- (furan-2-yl) acrylic acid (lib) in the presence of EDAC and DMAP in 1 ,2-dichloroethane to form the amide compound 12b. Basic hydrolysis of the ester methyl group present in compound 12b afforded compound 13.

Compound 13 along with the synthetic intermediate methyl esters 12a and 12b were evaluated for their inhibitory activity using the SI±AΛ₅₉ protein. Compound 13 did not show an enhancement of activity as expected. Instead compound 13 was found to be less active compared to compound 1 in the enzymatic assay (ICs₀ = 181 μM). However, the methyl ester intermediates (12a and 12b) showed improved inhibition as compared to corresponding acid derivatives 1 and 13. The methyl ester derivative of thiophene compound (12a) showed an IC₅₀ value of 71 μM and methyl ester derivative of furan compound (12b) showed an IC_5O value of 58 μM.

Compounds 14-17 were synthesized to evaluate the importance of the double bond present in compounds 1 and 13 for their inhibitory activity. Compounds 14 and 16 are the saturated analogs of the carboxylic acid derivatives 1 and 13, respectively. Compounds 15 and 17 are the saturated analogs of the methyl ester derivatives 12a and 12b, respectively. The SrtA_Λ59 protein assay showed that all four saturated compounds 14-17 were inactive up to a concentration of 600 μM, indicating that the double bond is necessary for the activity of the disclosed compounds. Synthesis of compounds 14 - 17 is outlined in Scheme 2 (FIG. 6).

Hydrogenation of the thiophene derivatives 1 and 12a using Pd/C as catalyst was not effective, possibly due to the catalyst poisoning effect of the thiophene ring present in these molecules. Use of a stronger catalyst like Pd black resulted in the hydrogenation of 1 and 12a to corresponding hydrogenated compounds 14 and 15. Hydrogenation of 13 and 12b were carried out under H₂ in the presence of Pd/C catalyst to afford compounds 16 and 17.

Several additional derivatives of compound 1 were created by incorporating different substituents in the place of the carboxylic acid group of compounds 1. Derivative included the following substitutions: CH₂OH (18a), CHO (19a) or CONH₂ (20). Substitution with CH₂OH and CHO groups did not result in a major change in the activity (18a, IC₅₀ = 73 μM and 19a, IC_5O ⁼ 77 μM), while substitution with CONH₂ group resulted in a decrease in activity as compared to compound 1 (20, IC₅₀ = 105 μM). Derivatives of the furan compound 13 were also created by incorporating different substituents in the place of the carboxylic acid group of compounds 13. Derivative included the following substitutions: CH₂OH (18b) and CHO (19b). These compounds showed improved inhibition as compared to parent furan compound 13. Compound 18b showed an ICs₀ value of 111 μM and compound 19b showed an IC₅₀ value of 107 μM. Synthesis of compounds 18a, 18b, 19a and 19b is outlined in Scheme 3 (FIG. 7). Compounds 12a and 12b were reduced using DIBAL in a mixture of anhydrous dichloromethane and THF to afford the alcohol derivatives 18a and 18b. Oxidation of alcohols using PCC in anhydrous THF afforded the aldehydes 19a and 19b. Synthesis of amide compound 20 is outlined in Scheme 4 (FIG. 8). Compound 20 was prepared from compound 1 by treatment with SOCl₂ followed by ammonia.

Synthesis of compounds 21-24 is outlined in Scheme 5 (FIG. 9). These compounds were designed to evaluate the nature of the linking group I on the activity of the compounds. Generally, the compounds containing triple bonds showed less activity than compounds containing double bonds. Compounds 21 and 22 showed IC₅₀ values of 165 and 183 μM, respectively (Table 2). Compounds containing a double bond in the cis configuration were more effective, with compounds 23 and 24 showing ICs₀ values of 61 and 154 μM, respectively (Table 2). The synthesis started from the known 3-(thiophen-2-yl)propiolic acid 35. The acid 35 was coupled with the amino compound 36 in the presence of EDAC and DMAP in 1,2- dichloroethane afforded the alkyne ester 21. The ester group present in compound 21 was hydrolyzed using NaOH in MeOH to afford the acetylenic acid 22. The partial hydrogenation of the acetylenic ester 21 using Lindlar Pd catalyst did not work as expected possibly due to the catalytic poisoning effect of the thiophene ring already present in the molecule. However, reduction worked smoothly to a cis double bonded product 23 when it was hydrogenated with H₂ in the presence of Pd/C in ethyl acetate. The ester group present in compound 23 was hydrolyzed using NaOH in MeOH to afford the cis acid 24.

Synthesis of compounds 25-28 is outlined in Scheme 6 shown in FIG. 10. These compounds were designed to evaluate substitutes at the at the nitrogen group. Generally, replacement of the H with a methyl group reduced the effectiveness of the compounds (IC₅₀ values fro compounds 25-28 were over 500 μM, Table 2). Compound 37 and 38 were methylated using methyl iodide in the presence of NaH in DMF to afford the N-methylated esters 25 and 26. Basic hydrolysis of 25 and 26 using IN. NaOH in MeOH afforded the final acid products 27 and 28, respectively.

Synthesis is of compound 29-30 is outlined in Scheme 7 shown in FIG. 11. These compounds were designed to evaluate the nature of the linking group I on the activity of the compounds. Compounds 29-30 contain a C3 alkenyl as the linking group as compared to a C₂ alkenyl linking group in the previous compounds. Generally, the compounds containing a C₃ alkenyl as the linking group I showed less activity than compounds containing a C₂ alkenyl as the linking group I. Compounds 29 and 30 showed IC₅₀ values of about 250 μM and greater (Table 2). Reductive amination of the known aldehyde 39 with the amine 36 in the presence of NaCNBH₃ and ZnCl₂ in methanol afforded the ester compound 29. Basic hydrolysis of compound 29 using NaOH in MeOH afforded the final acid product 30.

Synthesis of the compounds 31-34 is outlined in Scheme 8 shown in FIG. 12. These compounds were designed to evaluate substitution of the morpholine group the activity of the compounds. Compounds 31-34 contain a piperidine group rather than a morpholine group present in the previous compounds. Generally, the compounds containing a piperidine group showed similar activity to compounds containing a morpholine group. Compounds 31-34 showed IC₅₀ values of 92 μM, 131 μM, 181 μM and 463 μM, respectively (Table 2). Commercially available methyl 2-fluoro-5-nitro benzoate (40) was treated with piperidine in DMF to afford the compound 41, which upon hydrogenation using H₂ in the presence of Pd/C gave the amine 42. Compound 42 was coupled with trøn.s-3-(thiophene-2-yl) acrylic acid or trø«s-3-(furan-2-yl) acrylic acid in the presence of EDAC and DMAP in 1 ,2-dichloroethane to form the amides compound 31 or 32. Basic hydrolysis of the ester methyl group present in compound 31 and 32 afforded the compounds 33 and 34.

Synthesis of compounds 43-46 is outlined in Scheme 9 shown in FIG. 13. These compounds were designed to evaluate substitution of the compounds at the R₂ position on the activity of the compounds. Compounds 43-46 contain various alkyl and aralkyl substituents at the R₂ position in place of the esters and acids generally present on the parent compounds. Generally, the compounds 43-46 showed similar IC₅₀ values to previous compounds (see for example, 18a,b and 19a,b). Compounds 43-46 showed IC₅₀ values of 95 μM, 104 μM, 80 μM and 78 μM, respectively (Table 2). Compound 13 (1 mmol) was alkylated with alkyl bromides, R-Br (1.2 mmol) in the presence Of K₂CO₃ (2 mmol) in anhydrous N,N-dimethylformamide (5 mL) at room temperature for 12 h. TLC examination (50% EtOAc in CHCl₃) showed that the reaction is complete. The solvent was completely removed under high vacuum and the residue obtained was dissolved in EtOAc (20 mL). The solution was washed with IM. NaHCO₃ (2 x 15 mL), water (2 x 15 mL), brine (1 x 15 mL) and dried over anhydrous Na₂SO₄. The drying agent was filtered off and the solvent was evaporated to obtain the crude product. It was purified by column chromatography over silica gel using EtOAc / hexanes (1 :1) as eluent to afford the pure products 43-46.

Synthesis of compounds 51-56 is outlined in Scheme 10 shown in FIG. 14. These compounds were designed to evaluate substitution of the compounds at the position of the 5- membered (left most) ring on activity of the compounds. Compounds 51-56 contain benzyl and piperidine in place of thiophene and furan groups generally present on the parent compounds. Generally, the compounds 51-56 showed similar IC₅₀ values to previous compounds. Compounds 51-56 showed IC₅₀ values of 208 μM, 245 μM, >300 μM, n.d.,204 μM and 242 μM, respectively (Table 2). To a solution of 10 (1 mmol) in 1 ,2-dichloroethane (20 mL) aryl acrylic acid (1.2 mmol), EDAC (2 mmol), and DMAP (0.1 mmol) were added and stirred for 16 h at room temperature. TLC analysis (50% EtOAc in CHCl₃) revealed that the reaction is complete. The reaction mixture was diluted with 1 ,2-dichloroethane (50 mL) and the solution was washed with saturated NaHCO₃ solution (2 x 25 mL), water (1 x 25 mL), brine (1 x 25 mL) and dried over anhydrous Na₂SO₄. The drying agent was filtered off and the solvent was concentrated in vacuo to afford the crude product which was purified by column chromatography over silica gel using EtOAc / hexanes (1 : 1) as eluent to afford the pure products 51, 52 and 55. Compounds 51, 52 and 55 (1 mmol) in a mixture of THF (3 mL) and MeOH (3 mL) were treated with 3N. NaOH solution (2.5 mL) at room temperature and the resulting mixture was refluxed gently for 45 min. TLC analysis (50% EtOAc in CHCl₃) revealed that the reaction is complete. The solvent was completely removed in vacuo and the residue obtained was taken up in water (3 mL). The resulting mixture was cooled to 0 °C and acidified to pH ~2 by carefully adding 3N HCl. The resulting white solid was filtered and dried under vacuum to furnish pure compounds 53, 54 and 56.

Synthesis of compound 57 is outlined in Scheme 11 showin in FIG. 15. This compounds represents a new strucutral formula for sortase inhibitors different from the strcuture of the compounds in formulas Ia-Ic. Commercially available 4-hydroxy acetophenone (68) is refluxed with 4-chloronitrobenzene in the presence of KOH in DMF afforded the ether derivatice 69. Coupling of ether 69 with N,N-dimethylformamide dimethyl acetal in DMF afforded the compound 57.

Compounds 58, 60-63 and 67 are presented in Table 2. These compounds were generated as a result of the SAR studies discussed above. Compounds 58, 60-63 and 67 are represented by the general formula Ic and share structural similarity to the general formulas provided in structures Ia and Ib. Specifically, these compounds provide additional variation along the phenyl group to provide configurations similar to the morpholine and piperidine groups present in many of the previous compounds. Compounds 59, 64 and 66 also share similarity with the compounds discussed above in that they share a thiophene group linked to an N group via a C₂-C₃ alkenyl linkage containing at least one double bond. Although compounds 58-64 and 66- 67 have not yet been tested against sortase enzymes in vitro, they can be expected to display inhibitory activity based on the similarity to the compounds disclosed herein. Pharmaceutical Compositions and Modes of Administration

The present disclosure provides compounds of the general formula (I), (Ia), (Ib), (Ic) and II-XII and pharmaceutical compositions comprising the same. In a specific embodiment, the present disclosure provides compounds 12-13, 18-20, 23-24, 31-32 and 43-46 and pharmaceutical compositions comprising the same. In one embodiment, these compounds are useful as inhibitors of bacterial sortase enzymes. In a particular embodiment, these compounds are useful as inhibitors of bacterial SrtA, SrtB, SrtC and/or SrtD or homologs thereof. In a further particular embodiment, the compounds are useful as inhibitors of bacterial SrtA. The compounds of the present disclosure have utility in treating and/or preventing a variety of disease states and/or conditions involving bacterial infection and virulence, wherein said bacteria causing or contributing to the bacteria infection and virulence express a sortase enzyme.

The present disclosure provides for a pharmaceutical composition comprising a pharmaceutically effective amount of at least one compound of the present disclosure as described herein. Such pharmaceutical compositions may be used in the manufacture of a medicament for treating and/or preventing a disease or condition in which it is desirable to inhibit involving bacterial infection and virulence. Such pharmaceutical compositions and medicaments may also comprise a pharmaceutically acceptable carrier. The compounds of the disclosure are useful in both free form and in the form of pharmaceutically acceptable salts.

The pharmaceutically acceptable carriers described herein, including, but not limited to, vehicles, adjuvants, excipients, or diluents, are well-known to those who are skilled in the art. Typically, the pharmaceutically acceptable carrier is chemically inert to the active compounds and has no detrimental side effects or toxicity under the conditions of use. The pharmaceutically acceptable carriers can include polymers and polymer matrices.

The compounds described in the instant disclosure can be administered by any conventional method available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in combination with additional therapeutic agents.

The compounds described are administered in therapeutically effective amount. The therapeutically effective amount of the compound and the dosage of the pharmaceutical composition administered will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration, the age, health and weight of the recipient; the severity and stage of the disease state or condition; the kind of concurrent treatment; the frequency of treatment; and the effect desired. A daily dosage of active ingredient can be expected to be about 0.001 to 1000 milligrams (mg) per kilogram (kg) of body weight. In one embodiment, the total amount is between about 0.1 mg/kg and about 1000 mg/kg of body weight; in an alternate embodiment between about 1.0 mg/kg and about 100 mg/kg of body weight; in yet another alternate embodiment between 0.1 mg/kg and about 30 mg/kg of body weight. The above described amounts may be administered as a series of smaller doses over a period of time if desired. As would be obvious, the dosage of active ingredient may be given other than daily if desired.

The total amount of the compound administered will also be determined by the route, timing and frequency of administration as well as the existence, nature, and extent of any adverse side effects that might accompany the administration of the compound and the desired physiological effect. It will be appreciated by one skilled in the art that various conditions or disease states, in particular chronic conditions or disease states, may require prolonged treatment involving multiple administrations.

Dosage forms of the pharmaceutical compositions described herein (forms of the pharmaceutical compositions suitable for administration) contain from about 0.1 mg to about 500 mg of active ingredient (i.e. the compounds disclosed) per unit. In these pharmaceutical compositions, the active ingredient will ordinarily be present in an amount of about 0.5-95% weight based on the total weight of the composition. Multiple dosage forms may be administered as part of a single treatment. The active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups and suspensions. It can also be administered parenterally, in sterile liquid dosage forms. The active ingredient can also be administered intranasally (nose drops) or by inhalation via the pulmonary system, such as by nebulizers, propellant based metered dose inhalers or dry powders inhalation devices. Other dosage forms are potentially possible such as administration transdermally, via a patch mechanism or an ointment.

Formulations suitable for oral administration can consist of (a) liquid solutions, such as a pharmaceutically effective amount of the compound dissolved in diluents, such as water, saline, or orange juice; (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined pharmaceutically effective amount of the active ingredient, as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; and (e) suitable emulsions. Liquid formulations may include diluents, such as water and alcohols, for example, ethanol, benzyl alcohol, propylene glycol, glycerin, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch. Tablet forms can include one or more of the following: lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers. Lozenge forms can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acadia, emulsions, and gels containing, in addition to the active ingredient, such carriers as are known in the art.

Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain anti -oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the patient, and aqueous and non- aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The compound can be administered in a physiologically acceptable diluent in a pharmaceutically acceptable carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol, isopropanol, or hexadecyl alcohol, glycols, such as propylene glycol or polyethylene glycol such as poly(ethyleneglycol) 400, glycerol ketals, such as 2,2-dimethyl-l,3-dioxolane-4-methanol, ethers, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

Oils, which can be used in parenteral formulations include petroleum, animal, vegetable, or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral. Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters. Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents such as, for example, dimethyldialkylammonium halides, and alkylpyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene polypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl .beta.-aminopropionates, and 2-alkylimidazoline quaternary ammonium salts, and (e) mixtures thereof.

The parenteral formulations typically contain from about 0.5% to about 25% by weight of the active ingredient in solution. Suitable preservatives and buffers can be used in such formulations. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations ranges from about 5% to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol.

Pharmaceutically acceptable excipients are also well-known to those who are skilled in the art. The choice of excipient will be determined in part by the particular compound, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of the pharmaceutical composition of the present invention. The following methods and excipients are merely exemplary and are in no way limiting. The pharmaceutically acceptable excipients preferably do not interfere with the action of the active ingredients and do not cause adverse side-effects. Suitable carriers and excipients include solvents such as water, alcohol, and propylene glycol, solid absorbants and diluents, surface active agents, suspending agent, tableting binders, lubricants, flavors, and coloring agents.

The compounds of the present disclosure, alone or in combination with other suitable components, can be made into aerosol formulations to be administered via inhalation. These aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, and nitrogen. Such aerosol formulations may be administered by metered dose inhalers. They also may be formulated as pharmaceuticals for non-pressured preparations, such as in a nebulizer or an atomizer.

The formulations can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient, for example, water, for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets. The requirements for effective pharmaceutically acceptable carriers for injectable compositions are well known to those of ordinary skill in the art. See Pharmaceutics and Pharmacy Practice, J.B. Lippincott Co., Philadelphia, Pa., Banker and Chalmers, Eds., 238- 250 (1982) and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., 622-630 (1986).

Formulations suitable for topical administration include pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, as well as creams, emulsions, and gels containing, in addition to the active ingredient, such carriers as are known in the art.

Additionally, formulations suitable for rectal administration may be presented as suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases. Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulas containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate.

One skilled in the art will appreciate that suitable methods of administering a compound of the present invention to an patient are available, and, although more than one route can be used to administer a particular compound, a particular route can provide a more immediate and more effective reaction than another route. Methods of Treatment and Prevention

In one embodiment, the teachings of the present disclosure provide for the use of a compound of the present disclosure or pharmaceutical compositions containing a compound of the present disclosure in a method of treating a bacterial infection or a disease state and/or condition which is caused by, at least in part, bacterial infection and/or virulence. In one embodiment, such disease states and conditions include, but are not limited to, hospital-acquired bacterial infections, endocarditis, osteomyelitis, septic arthritis pneumonia, toxic shock syndrome, skin infections, meningitis, anthrax, B. cereus food poisoning, tetanus, clostridial myonecrosis, pneumonia, toxic shock syndrome, skin infections, meningitis, ear infections, strep throat, rheumatic fever, scarlet fever, puerperal fever, listeriosis, perinatal septicemia, encephalitis, intrauterine infections sinusitis and severe invasive infections; however, other disease states and conditions may be treated as would be obvious to one of ordinary skill in the art.

The method of treatment comprises the steps of providing such composition or pharmaceutical composition and administering such compound or pharmaceutical composition in a therapeutically effective amount to inhibit or reduce bacterial infection and/or virulence in a patient in need of such treatment. One or more compounds of the present disclosure may be used or the pharmaceutical composition may contain one or more compounds of the present disclosure. By decreasing, at least in part, the activity of a bacterial sortase enzyme, the compounds of the present disclosure reduce the ability of the bacteria to infect the cells of the subject and/or reduce the virulence of the bacteria towards the host. Furthermore, the viability of the bacteria is also reduced. As a result, the disease state and/or condition which is caused by, at least in part, bacterial infection and/or virulence is thereby treated in the subject. In an alternate embodiment, the teachings of the present disclosure provide for the use of a compound of the present disclosure or pharmaceutical compositions containing a compound of the present disclosure in a method of preventing or suppressing a bacterial infection or a disease state and/or condition which is caused by, at least in part, bacterial infection and/or virulence. In one embodiment, such disease states and conditions include, but are not limited to, hospital- acquired bacterial infections, endocarditis, osteomyelitis, septic arthritis pneumonia, toxic shock syndrome, skin infections, meningitis, anthrax, B. cereus food poisoning, tetanus, clostridial myonecrosis, pneumonia, toxic shock syndrome, skin infections, meningitis, ear infections, strep throat, rheumatic fever, scarlet fever, puerperal fever, listeriosis, perinatal septicemia, encephalitis, intrauterine infections sinusitis and severe invasive infections; however, other disease states and conditions may be treated as would be obvious to one of ordinary skill in the art.

The method of preventing or suppressing comprises the steps of providing such compound or pharmaceutical composition and administering such compound or pharmaceutical composition in a therapeutically effective amount to inhibit or reduce bacterial infection and/or virulence in a patient in need of such preventing or suppressing. One or more compounds of the present disclosure may be used or the pharmaceutical composition may contain one or more compounds of the present disclosure. By decreasing, at least in part, the activity of a bacterial sortase enzyme, the compounds of the present disclosure reduce the ability of the bacteria to infect the cells of the subject and/or reduce the virulence of the bacteria towards the host. Furthermore, the viability of the bacteria is also reduced. As a result, the disease state and/or condition which is caused by, at least in part, bacterial infection and virulence is thereby prevented or suppressed in the subject.

In a specific embodiment, the disease state or condition caused, at least in part, by a bacterial infection or virulence is endocarditis. Endocarditis may be related infection of the endocardium and/or the heart valves by bacteria. Staphylococcus aureus is a common cause of endocarditis. Through inhibition of the sortase enzyme, such as, but not limited to SrtA, the ability of bacteria, such as, but not limited to, Staphylococcus aureus, to bind to and infect the endocardium and/or heart valves may be reduced; furthermore, pre-existing bacterial infections may be eliminated. The foregoing leads to a decrease in disease. In yet another alternate embodiment, the present disclosure provides for the use of a compound of the present disclosure or pharmaceutical compositions containing a compound of the present disclosure in methods of reducing or eliminating bacterial virulence in a subject in need of such reduction. The method of reducing or eliminating comprises the steps of providing such compound or pharmaceutical composition and administering such compound or pharmaceutical composition in a therapeutically effective amount to reduce or eliminate bacterial virulence in a patient in need of such reducing or eliminating. One or more compounds of the present disclosure may be used or the pharmaceutical composition may contain one or more compounds of the present disclosure. By reducing or eliminating, at least in part, the activity of a bacterial sortase enzyme, the compounds of the present disclosure reduce the virulence of the bacteria towards the host. As a result, health of the subject is improved.

In yet a further alternate embodiment, the present disclosure, provides for the use of a compound of the present disclosure or pharmaceutical compositions containing a compound of the present disclosure in methods of treating bacterial infection.

The method of treatment comprises the steps of providing such composition or pharmaceutical composition and administering such compound or pharmaceutical composition in a therapeutically effective amount to inhibit or reduce bacterial infection in a patient in need of such treatment. One or more compounds of the present disclosure may be used or the pharmaceutical composition may contain one or more compounds of the present disclosure. By decreasing, at least in part, the activity of a bacterial sortase enzyme, the compounds of the present disclosure reduce the ability of the bacteria to infect the cells of the subject an/or reduce the virulence of the bacteria towards the host. Furthermore, the viability of the bacteria is also reduced. As a result, the bacterial infection is thereby treated in the subject. In still a further alternate embodiment, the present disclosure provides for the use of a compound of the present disclosure or pharmaceutical compositions containing a compound of the present disclosure in methods of preventing or suppressing bacterial infection.

The method of preventing or suppressing comprises the steps of providing such compound or pharmaceutical composition and administering such compound or pharmaceutical composition in a therapeutically effective amount to inhibit or reduce bacterial infection in a patient in need of such preventing or suppressing. One or more compounds of the present disclosure may be used or the pharmaceutical composition may contain one or more compounds of the present disclosure. By decreasing, at least in part, the activity of a bacterial sortase enzyme, the compounds of the present disclosure reduce the ability of the bacteria to infect the cells of the subject an/or reduce the virulence of the bacteria towards the host. Furthermore, the viability of the bacteria is also reduced. As a result, the bacterial infection is thereby prevented or suppressed in the subject.

In the foregoing methods of use, the bacteria may be any bacteria or other organism that contains and expresses a sortase enzyme, hi one embodiment, the sortase enzyme is SrtA, SrtB, SrtC, and/or SrtD, and/or homologues of any of the foregoing. In a specific embodiment, the sortase enzyme is SrtA. In another embodiment, the bacteria is Staphylococcus aureus. Furthermore, the bacteria may one that is resistant to a commonly used antibiotic, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococcus (VRE), vancomycin-intermediate Staphylococcus aureus (VISA) and vancomycin-resistant Staphylococcus aureus (VRSA). The bacteria may be Gram-positive bacteria, Gram-negative bacteria or combinations of the foregoing that expresses a sortase enzyme; in one embodiment, the bacteria is a Gram-positive bacteria that expresses the sortase enzyme. Representative bacteria that have been documented to express a sortase enzyme, include, but are not limited to, Acidothermus cellulolyticus, Actinomyces naeslundii, Actinomyces viscosus, Arthrobacter aurescens, Arthrobacter sp. FB24, Bacillus anthracis, Bacillus cereus, Bacillus clausii, Bacillus halodurans, Bacillus licheniformis, Bacillus subtilis, Bacillus thuringiensis, Bacillus weihenstephanensis, Bifidobacterium longum, Bradyrhizobium japonicum, Chloroflexus aurantiacus, Clostridium acetobutylicum, Clostridium difficile.Clostridium limosum, Clostridium perfringens, Clostridium tetani, Congregibacter litoralis, Corynebacterium diphtheriae, Corynebacterium efficiens, Corynebacterium glutamicum, Corynebacterium jeikeium, Desulfitobacterium hafniense, Enterococcus faecalis, Enterococcus faecium, Exiguobacterium sibiricum, Exiguobacterium sp. 255-15, Frankia sp. CcB, Fran/da sp. EAN] pec, Geobacillus kaustophilus, Hahella chejuensis, Kineococcus radiotolerans, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus sakei, Lactobacillus salivarius, Lactococcus lactis, Leifsonia xyli, Leuconostoc mesenteroides, Listeria innocua, Listeria monocytogenes, Listeria welshimeri, Marinobacter hydrocarbonoclasticus, Nocardioides sp. JS614, Oceanobacillus iheyensis, Pediococcus pentosaceus, Pseudoalteromonas atlantica, Pseudomonas mendocina, Rhodobacter sphaeroides, Rubrobacter xylanophilus, Saccharophagus degradans, Shewanella amazonensis, Shewanella baltica, Shewanella frigidimarina, Shewanella oneidensis, Shewanella sp. ANA-3, Shewanella sp. MR-4, Shewanella sp. MR-7, Shewanella sp. W3-18-1, Solibacter usitatus, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus gordonii, Streptococcus mutans, Streptococcus oralis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus sanguinis, Streptococcus suis, Streptococcus thermophilus, Streptomyces avermitilis, Streptomyces coelicolor, Symbiobacterium thermophilum, Thermobifida fusca and Tropheryma whipplei. Representative other organisms that have been demonstrated to contain a sortase enzyme include, but are not limited to, Methanothermobacter thermautotrophicus. However, any bacteria or organism that expresses a sortase enzyme may be subject to the methods of the present disclosure.

Furthermore, the methods of use described herein may further comprise the administration of a therapeutically effective amount of one or more additional agents to further improve the efficacy of the methods described herein. The additional agents may be administered at the same time or a different time than a compound of the present disclosure. In one embodiment, the additional agent has a mechanism of action that is distinct from the compounds of the present disclosure (i.e., the second compound is not an inhibitor of a sortase enzyme). As the mechanisms of action the additional agent and the compounds of the present disclosure are different, the combination of the compounds of the present disclosure and the additional agent are expected to be effective in the methods described herein. The identity and nature of the additional agent may be determined by the healthcare provider based on his/her knowledge of the subject's condition.

In one embodiment, the additional agent is an antibiotic. Any antibiotic or antibiotic combinations may be used as the additional agent. As the compounds of the present disclosure operate in a manner that is different and distinct from the mechanisms of action of currently known antibiotics, the combination of the compounds of the present disclosure and antibiotics are expected to be effective in the methods described herein. Representative antibiotics may be selected from the classes of aminoglycosides, ansamycins, carbacephem, carbapenems, cephalosporins, glycopeptides, macrolides, monobactams, penicillins, polypeptides, quinolones, sulfonamides and/or tetracyclines; other classes of antibiotics may also be used. Exemplary antibiotics include, but are not limited to, Amikacin, Gentamicin, Kanamycin, Neomycin, Netilmicin, Streptomycin, Tobramycin, Geldanamycin, Herbimycin, Loracarbef, Ertapenem, Imipenem/Cilastatin, Meropenem, Cefadroxil, Cefazolin, Cephalexin, Cefaclor, Cefamandole, Cefoxitin, Cefprozil, Cefuroxime, Cefixime, Cefdinir, Cefditoren, Cefoperazone, Cefotaxime, Cefpodoxime, Ceftazidime, Ceftibuten, Ceftizoxime, Ceftriaxone, Cefepime, Teicoplanin, Vancomycin, Methicillin, Azithromycin, Clarithromycin, Dirithromycin, Erythromycin, Roxithromycin, Troleandomycin, Aztreonam, Amoxicillin, Ampicillin, Azlocillin, Carbenicillin, Cloxacillin, Dicloxacillin, Flucloxacillin, Mezlocillin, Nafcillin, Penicillin, Piperacillin, Ticarcillin, Bacitracin, Trimethoprim Colistin, Polymyxin B, Ciprofloxacin, Enoxacin, Gatifloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Norfloxacin, Ofloxacin, Trovafloxacin, Mafenide, Prontosil, Sulfacetamide, Tobramycin Sulfamethizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole, Trimethoprim, Trimethoprim-Sulfamethoxazole (Co-trimoxazole) (TMP-SMX), Demeclocycline, Doxycycline, Minocycline, Oxytetracycline, Tetracycline, Arsphenamine, Chloramphenicol, Clindamycin, Ethambutol, Fosfomycin, Fusidic acid, Furazolidone, Isoniazid, Linezolid, Metronidazole, Mupirocin, Nitrofurantoin, Platensimycin, Pyrazinamide, Quinupristin/Dalfopristin, Rifampin, Spectinomycin and/or Telithromycin. In a specific embodiment, the antibiotic is methicillin and/or vancomycin. In this embodiment, the co-administration of the foregoing agents with a sortase inhibitor of the present disclosure increases the efficacy of the foregoing antibiotics against their bacterial targets.

The foregoing description illustrates and describes the compounds of the present disclosure. Additionally, the disclosure shows and describes only certain embodiments of the compounds but, as mentioned above, it is to be understood that the teachings of the present disclosure are capable of use in various other combinations, modifications, and environments and is capable of changes or modifications within the scope of the inventive concept as expressed herein, commensurate with the above teachings and/or the skill or knowledge of the relevant art. The embodiments described hereinabove are further intended to explain best modes known of practicing the invention and to enable others skilled in the art to utilize the invention in such, or other, embodiments and with the various modifications required by the particular applications or uses of the invention. Accordingly, the description is not intended to limit the invention to the form disclosed herein. The claim below is appended for purposes of foreign priority only, if required. All references cited herein are incorporated by reference as if fully set forth in this disclosure. METHODS

Fluorescence Resonance Energy Transfer (FRET^") assay

To determine IC₅₀ values each potential inhibitor, the previously published fluorescenceresonance energy transfer (FRET) assay was used. Kruger, R. G.; Dostal, P.; McCafferty, D. G., Development of a high-performance liquid chromatography assay and revision of kinetic parameters for the Staphylococcus aureus sortase transpeptidase SrtA. Anal Biochem 2004, 326, (1), 42-8; Ton-That, H.; Mazmanian, S. K.; Faull, K. F.; Schneewind, O., Anchoring of surface proteins to the cell wall of Staphylococcus aureus. Sortase catalyzed in vitro transpeptidation reaction using LPXTG peptide and NH(2)-Gly(3) substrates. J Biol Chem 2000, 275, (13), 9876-81. This FRET assay employs the use of the donor and quencher pair, EDANS and Dabcyl, respectively. The peptide Dabcyl-QALPETGEE-EDANS was purchased from a commercial source. The EDANS fluorophore has an excitation wavelength, λ_ex>d = 336 nm and an emission maximum, λ_emjd = 490 nm. The fluorescence emission spectra for EDANS overlaps very well with the absorption spectra of Dabcyl, where Dabcyl has an absorption maximum, λ_max,_a = 472 nm. As such, when the two molecules are spatially close, as they are in the intact peptide, the efficiency of transfer from EDANS to Dabcyl is high. Upon excitation of EDANS little emission from the donor is observed because most of the energy from fluorescence is transferred to the dabcyl acceptor and lost through non radiative decay pathways. In contrast, upon cleavage the newly formed fragments will diffuse apart and the two peptide fragments will become spatially separated and the FRET efficiency will diminish. Thus, the emission from the EDANS fluorophore will precipitously increase. Because of these properties, this FRET pair has been successfully employed to monitor SrtA catalyzed peptide cleavage.

Traditionally, this assay is performed in a fluorometer under steady-state or multiple turnover conditions, where steady-state conditions are defined as the substrate being in large excess over the enzyme. In this experimental design a constant enzyme concentration is titrated with increasing concentrations of the peptide substrate. The EDANS fluorophore is excited at 350 nm and fluorescence from EDANS is observed at 495 nm over a period of time. To reduce the correction for the inner filter effect the reaction was monitored at an emission of 590 nm. At the beginning of the observation, fluorescence is low due to a high FRET efficiency, but, as the peptide is cleaved the fluorescence increases linearly. The slope of this linear increase is taken to be the initial velocity, V₀. In principle, the initial velocity as a function of peptide concentration will exhibit Michaelis-Menten kinetics, i.e. a hyperbolic dependence on substrate concentration, and can be analyzed to yield the steady-state kinetic constants K_n, and k_cat. Because of the large inner filter correction at high concentrations of substrate, the effect of inhibitors on V₀ was monitored at select enzyme and substrate concentration. These experiments are performed by mixing 5 μM SrtA with 20 μM Dabcyl-EDANS labeled peptide in a 0.5 cm cuvette. This mixture is excited at λ_ex = 350 nm and fluorescence is observed at λ_em = 590 nm. From this experiment, a time course of fluorescence increase as a function of time is acquired. The slope of this line is considered the maximum velocity in the absence of inhibitor. The identical experiment is then performed in the presence of increasing concentrations of inhibitor and the slopes are compared until a 50 % decrease in the slope is observed. The concentration of inhibitor where 50 % of this maximum velocity is observed is taken to be the IC₅₀. Each experiment was repeated at least three times to ensure the reproducibility and calculate standard deviation of the reported IC₅₀ values. General Methods for Synthesis

Solvent evaporations were carried out in vacuo with a rotary evaporator. Analytical samples were prepared by drying the samples in vacuo (0.2 mmHg) in an Abderhalden drying apparatus over P₂O₅ and ethyl acetate at reflux temperature. Thin layer chromatography (TLC) was performed on silica gel plates with fluorescent indicator (Whatmann, silica gel, UV254, 25 μm plates). Spots were visualized by UV light (254 and 365 ran). AU analytical samples were single spots on TLC in at least two different solvent systems. Purification by column and flash chromatography was carried out using 'BAKER' silica gel (40 μm) in the solvent systems indicated. The amount (weight) of silica gel for column chromatography was in the range of 50- 100 times the amount (weight) of the crude compounds being separated. Melting points were determined on a Mel-Temp II melting point apparatus and are uncorrected. Proton nuclear magnetic resonance (¹H NMR) and carbon nuclear magnetic resonance (¹³C NMR) spectra were recorded on a Brucker DPX-300 spectrometer using TMS as internal standard. The values of chemical shifts (δ) are given in ppm and coupling constants (J) in Hz. The chemical shift values are reported as parts per million (ppm) relative to tetramethylsilane as internal standard. Mass spectra were recorded on a Micromass Platform LCC instrument. Elemental analyses were performed by Atlantic Microlab, Norcross, Georgia and the results indicated by symbols for the elements were within ±0.4% of theoretical values. Anhydrous solvents used for reactions were purchased in Sure-Seal™ bottles from Aldrich Chemical Company. Other reagents were purchased from Aldrich, Lancaster or Acros chemical companies and used as received. Methyl 5-amino-2-morpholinobenzoate (Compound 10)

To a solution of Methyl 5-nitro-2-morpholinobenzoate 9 (1.0 g, 3.76 mmol) in EtOAc (35 mL) 10% PdVC (100 mg) was added and stirred under an atmosphere of H₂ from a balloon (~1 atm) for 12 h. TLC analysis (1 :1 EtOAcZCHCl₃) revealed that the reaction was complete. The catalyst was removed by filtration through a celite bed and the filtrate was concentrated on vacuum to furnish 10 as a solid (0.88 g, 99 %): mp 121 - 122 ⁰C; ¹H NMR (CDCl₃) δ 2.91 - 2.97 (m, 4H), 3.60 (bs, 2H), 3.79 - 3.85 (m, 4H), 3.87 (s, 3H), 6.78 (dd, IH, J₁ = 3.0 Hz, J₂ = 8.7 Hz), 6.95 (d, IH, J = 8.4 Hz) and 7.05 (d, IH, J = 2.7 Hz); ¹³C NMR (CDCl₃) δ 51.7, 53.3, 67.1, 116.7, 118.8, 120.8, 127.0, 141.9, 143.7 and 168.0; MS (ES) m/z 237 (M + H); Anal. (C₁₂H₁₆N₂O₃) C₅ H₅ N. Methyl 5-((EV3-(thiophen-2-yl)acrylaniido)-2-rnorpholinobenzoate (Compound 12a) To a solution of 10 (0.430 g, 1.82 mmol) in 1 ,2-dichloroethane (20 mL) 3-(2- thienyl)acrylic acid (0.337 g, 2.18 mmol), EDAC (1.12 g, 5.82 mmol), and DMAP (0.022 mg, 0.182 mmol) were added and stirred for 16 h at room temperature. TLC analysis (1 :1 EtOAc/CHCl₃) revealed that the reaction was complete. The reaction mixture was diluted with dichloromethane (50 mL) and the organic layer was washed with saturated NaHCO₃ solution (2 x 25 mL), water (1 x 25 mL), brine (1 x 25 mL) and dried over anhydrous Na₂SO₄. The drying agent was filtered off and the solvent was concentrated in vacuo to afford the crude product which was crystallized form a mixture of CHCl₃ and hexanes to furnish 12a as a yellow solid (0.62 g, 91%): mp 196 - 197 ⁰C; ¹H NMR (CDCl₃) δ 3.00 - 3.05 (m, 4H), 3.83 - 3.88 (m, 4H), 3.89 (s, 3H), 6.31 (d, IH, J = 15.3 Hz), 7.03 - 7.08 (m, 2H), 7.24 - 7.28 (m, 2H), 7.33 - 7.38 (m,lH), 7.80 (d, IH, J = 8.7 Hz), 7.83 - 7.90 (m, IH) and 7.92 (d, IH, J = 2.3 Hz); ¹³C NMR (CDCl₃) δ 52.2, 53.1, 67.2, 1 19.3, 119.8, 123.2, 124.6, 124.9, 127.9, 128.2, 130.9, 132.5, 135.1, 139.8, 149.0, 163.7 and 167.5; MS (ES) m/z 373 (M + H); Anal. (Ci₉H₂₀N₂O₄S) C, H, N. Methyl 5-((iπ-3-(furan-2-y0acrylamido)-2-morpholinobenzoate (Compound 12b) Compound 12b was synthesized, following a similar procedure as the one used for preparation of 12a starting from compound 10 (0.426 g, 1.8 mmol), 3-(2-furyl)acrylic acid (0.298 g, 2.16 mmol), EDAC (1.10 g, 5.76 mmol), and DMAP (0.022 mg, 0.182 mmol) to obtain the pure product (0.639 g, 99 %): mp 172 - 173 °C; ¹H NMR (CDCl₃) δ 2.98-3.07 (m, 4H), 3.82-3.88 (m, 4H), 3.89 (s, 3H), 6.42 (d, IH, J = 15.0 Hz), 6.48 (dd, IH, J₁ = 1.8 Hz, J₂ = 3.3 Hz), 6.60 (d, IH, J = 3.6 Hz), 7.04 (d, IH, J = 9.0 Hz), 7.40 (bs, IH), 7.46 (d, IH, J = 1.5 Hz), 7.52 (d, IH, J = 15.0 Hz), 7.82 ( d, IH, J= 7.2 Hz) and 7.91 (s, IH); ¹³C NMR (CDCl₃) δ 52.1, 53.0, 67.1, 112.3, 114.4, 118.3, 119.6, 123.2, 124.6, 128.8, 132.7, 144.2, 148.8, 151.1 (2C), 164.1 and 167.6; MS (ES) m/z 357 (M + H); Anal. (C₁₉H₂₀N₂O₅) C, H, N. 5-((E)-3-(Thiophen-2-yl)acrylamido)-2-morpholinobenzoic acid (Compound 1) To a solution of compound 12a (0.300 g, 0.81 mmol) in a mixture of THF (1.5 mL) and methanol (1.5 mL), 3N. NaOH solution (2.5 mL) was added at room temperature and the resulting mixture was refluxed gently for 45 min. TLC analysis (1 :1 ΕtOAc/CHCl₃) revealed that the reaction was complete. The solvent was completely removed in vacuo and the residue obtained was taken up in water (3 mL). The resulting mixture was cooled to 0 ⁰C and acidified carefully with 3N HCl (-2.5 mL). The resulting white solid was filtered and dried under vacuum to furnish pure compound 1 (0.283 g, 98%): mp 278-279 °C; ¹H NMR (DMSO-d₆) δ 3.00-3.09 (m, 4H), 3.74-3.85 (m, 4H), 6.55 (d, IH, J = 15.3 Hz), 7.14 (dd, IH, J, = 3.6 Hz, J₂ = 5.1 Hz), 7.47 (d, IH, J = 3.3 Hz), 7.67 (s, IH), 7.68 (d, IH, J = 3.0 Hz), 7.77 (d, IH, J = 15.3 Hz), 7.99 (dd, IH, J₁ = 3.6 Hz, J₂ = 5.1 Hz), 8.29 (d, IH, J= 2.7 Hz) and 10.4 (bs, IH); ¹³C NMR (DMSO- d₆) δ 52.7, 66.3, 120.3, 120.9, 123.8, 124.0, 125.4, 128.5, 128.7, 131.6, 133.7, 137.8, 139.6, 145.2, 163.4 and 166.4; MS (ES) m/z 357 (M- H); Anal. (Ci₈H₁₈N₂O₄S) C, H, N. 5-((E)-3-(Furan-2-yl)acrylamido)-2-moφholinobenzoic acid (Compound 13)

Compound 13 was synthesized following a similar procedure as the one used for preparation of compound 1 starting from 12b (0.200 g, 0.57 mmol) and 3N NaOH solution to afford the pure product as a white solid (0.192 g, 99%): mp 281-282 °C; ¹H NMR (DMSO-d₆) δ 3.01-3.11 (m, 4H), 3.75-3.86 (m, 4H), 6.59 (d, IH, J = 15.3 Hz), 6.60-6.69 (m, IH), 6.87 (d, IH, J = 3.6 Hz), 7.41 (d, IH, J = 15.6 Hz), 7.68 (d, IH, J = 9.0 Hz), 7.84 (d, IH, J = 1.5 Hz), 8.00 (dd, IH, J₁ = 2.7 Hz, J₂ = 8.7 Hz), 8.29 (d, IH, J = 2.7 Hz) and 10.4 (bs, IH); ¹³C NMR (DMSO-d₆) δ 52.8, 66.4, 112.5, 115.0, 118.9, 120.9, 123.8, 124.0, 125.4, 127.8, 137.8, 145.2, 145.4, 150.8, 163.6 and 166.4; MS (ES) m/z 341 (M - H); Anal. (C₁₈H₁₈N₂O₅) C, H, N. 5-(3-(Thiophen-2-yl)propanamido)-2-morpholinobenzoic acid (Compound 14)

A solution of compound 1 (0.071 g, 0.20 mmol), ammonium formate (0.188 g, 2.97 mmol) and Pd black (40 mg) in a mixture of methanol (4 mL) and EtOAc (3 mL) was refluxed for 6 h. TLC analysis (1 :1 EtOAc/CHCl₃) and mass spectral data revealed that the reaction was complete. The catalyst was removed by filtration through a celite bed, washed with a mixture of methanol and CHCl₃, and the filtrate was concentrated under vacuum to afford the crude product. The crude product was purified by washing thoroughly with water to remove the inorganic salts, filtered and dried under vacuum to furnish pure product 14 (0.049 g, 69%): mp 226 - 227 °C; ¹H NMR (DMSO-d₆) δ 2.68 (t, 2H, J= 7.5 Hz), 3.13 (t, 2H, J= 7.5 Hz), 2.99 - 3.08 (m, 4H), 3.74 - 3.86 (m, 4H), 6.86 - 6.97 (m, 2H), 7.30 (dd, IH, J, = 1.0 Hz, J₂ = 4.9 Hz), 7.65 (d, IH, J= 8.4 Hz), 7.90 (dd, IH, J₁ = 2.7 Hz, J₂ = 8.7 Hz), 8.21 (d, IH, J = 2.4 Hz) and 10.2 (bs, IH); ¹³C NMR (DMSO-d₆) δ 24.9, 38.0, 52.7, 66.3, 120.8, 123.7, 123.8, 123.9, 124.7, 125.2, 126.9, 137.7, 143.4, 145.0, 166.3 and 170.1; MS (ES) m/z 361 (M + H); Anal. (Ci₈H₂₀N₂O₄S) C, H, N. Methyl 5-(3-(thiophen-2-yl)propanamido)-2-morpholinobenzoate (Compound 15)

A solution of compound 12a (0.100 g, 0.27 mmol), ammonium formate (0.255 g, 4.05 mmol) and Pd black (50 mg) in methanol (5 mL) was refluxed for 6 h. TLC analysis (1 :1 EtOAc/CHCl₃) and mass spectrum revealed that the reaction was complete. The catalyst was removed by filtration through a celite bed, washed with chloroform, and the filtrate was concentrated under vacuum. The residue obtained was dissolved in CH₂Cl₂ (20 mL) and water (15 mL) was added. The organic layer was separated and washed with saturated NaHCO₃ solution (2 x 15 mL), water (2 x 15 mL), brine (1 x 15 mL) and dried (Na₂SO₄). The drying agent was filtered off and the solvent was removed under vacuum to obtain the crude product. This crude product was purified by column chromatography over Si gel (4 x 2 cm) using 1 :5 EtOAc/CHCl₃ as eluent to afford the pure compound 15 (0.082 g, 82 %): mp 115 - 116 "C; ¹H NMR (CDCl₃) δ 2.70 (t, 2H, J = 7.5 Hz), 2.97 - 3.05 (m, 4H), 3.27 (t, 2H, J = 7.2 Hz), 3.81 - 3.87 (m, 4H), 3.88 (s, 3H), 6.86 (dd, IH, J₁ = 0.9 Hz, J₂ = 3.3 Hz), 6.90 - 6.96 (m, IH), 7.01 (d, IH, J= 9.0 Hz), 7.11 (bs, IH), 7.14 (dd, IH, J₁ = 1.2 Hz, J₂ = 5.1 Hz), 7.64 (dd, IH, J₁ = 2.7 Hz, J₂ = 8.7 Hz) and 7.76 (d, IH, J = 8.7 Hz); ¹³C NMR (CDCl₃) δ 25.5, 38.9, 52.0, 52.9, 67.0, 119.5, 123.2, 123.4, 124.6, 124.7 (2C), 126.8, 132.3, 143.0, 148.6, 167.4 and 170.1. MS (ES) m/z 375 (M + H). Anal. (Ci₉H₂₂N₂O₄S-O^SH₂O) C, H, N. 5-(3-(Furan-2-yl)propanamido)-2-morpholinobenzoic acid (Compound 16)

To a solution of compound 13 (0.075 g, 0.22 mmol) in EtOAc (15 mL) 10% Pd/C (20 mg) was added and stirred under a hydrogen atmosphere from balloon (~1 atm) for 25 min. TLC analysis (1 :1 EtOAc: CHCl₃) and ¹H NMR of aliquot revealed that the reaction was complete. (Note: Prolonged reaction times lead to hydrogenation of furan ring). The catalyst was removed by filtration through a celite bed and the filtrate was concentrated on vacuum to afford the crude product which was purified by column chromatography over Si gel (4 x 2 cm) using EtOAc as eluent to afford pure compound 16 as a white solid (0.066 g, 88 %): mp 230 - 231 °C; ¹H NMR (DMSO-d₆) δ 2.65 (t, 2H, J= 7.5 Hz), 2.93 (t, 2H, J= 7.3 Hz), 3.00-3.07 (m, 4H), 3.73-3.83 (m, 4H), 6.11 (dd, IH, J₁ = 1.2 Hz, J₂ = 2.1 Hz), 6.34 (dd, IH, J₁ = 1.8 Hz, J₂ = 3.0 Hz), 7.51 - 7.52 (m, IH), 7.65 (d, IH, J = 8.7 Hz), 7.89 (dd, IH, J₁ = 2.7 Hz, J₂ = 8.7 Hz), 8.21 (d, IH, J = 2.7 Hz) and 10.2 (bs, IH); ¹³C NMR (DMSO-d₆) δ 23.2, 34.5, 52.8, 66.4, 105.2, 1 10.4, 120.8, 123.8, 123.9, 125.3, 137.8, 141.5, 145.0, 154.4, 166.4 and 170.2; MS (ES) m/z 343 (M - H); Anal. (C₁₈H₂₀N₂O₅) C, H, N. Methyl 5-(3-(furan-2-yl)propanamido)-2-morpholinobenzoate (Compound 17)

Compound 17 was synthesized, following procedure used for the preparation of compound 16 starting from compound 12b (0.136 g, 0.38 mmol) in EtOAc (15 mL) and 10% Pd/C (20 mg) under a hydrogen atmosphere (0.106 g, 77 %): mp 119 - 120 °C; ¹H NMR (CDCl₃) δ 2.69 (t, 2H, J = 7.5 Hz), 2.97-3.03 (m, 4H), 3.07 (t, 2H, J = 7.3 Hz), 3.81-3.87 (m, 4H), 3.88 (s, 3H), 6.07 (d, IH, J= 3.3 Hz), 6.27-6.32 (m, IH), 7.01 (d, IH, J= 9.0 Hz), 7.16 (bs, IH), 7.32 (d, IH, J = 1.2 Hz), 7.65 (dd, IH, J₁ = 2.7 Hz, J₂ = 8.7 Hz) and 7.78 (d, IH, J = 2.4 Hz); ¹³C NMR (CDCl₃) δ 23.7, 35.4, 52.0, 52.9, 67.1, 105.5, 110.2, 119.5, 123.2, 124.6, 124.7, 132.3, 141.1, 148.7, 154.0, 167.5 and 170.3; MS (ES) m/z 357 (M - H). Anal. (Ci₉H₂₂N₂O₅O^SH₂O) C, H, N.

(EVN-(3-(Hvdroxymethyl)-4-moφholinophenyl)-3-(thiophen-2-vDacrylamide (Compound 18a) To a solution of compound 12a (0.500 g, 1.34 mmol) in a mixture of anhydrous CH₂Cl₂ (35 mL) and THF (5 mL), DIBAL-H (6.7 mL, IM in dichloromethane, excess) was added and stirred at 0 °C for 1 h. Then it was allowed to attain room temperature at which it was stirred for an additional 2 h. TLC analysis (1 :1 EtOAc/CHCl₃) revealed that the reduction was complete. The reaction mixture was cooled to 0 °C and was quenched by careful addition of methanol (3 mL) and IN. HCl (0.5 mL). The solvent was removed under reduced pressure and the residue obtained was dissolved in CHCl₃ (300 mL) and washed with water (2 x 75 mL), brine (1 x 150 mL) and dried (Na₂SO₄). The drying agent was filtered off and the solvent was removed under vacuum to afford the crude product which was purified by flash column chromatography on silica gel (20 x 3 cm) using EtOAc/hexanes (3: 1) as eluent to furnish compound 18a as a yellow solid (0.354 g, 77 %): mp 184 - 185 °C; ¹H NMR (DMSOd₆) δ 2.79 - 2.83 (m, 4H), 3.65 - 3.75 (m, 4H), 4.54 (d, 2H, J- 5.1 Hz), 5.12 (t, IH, J= 5.4 Hz), 6.58 (d, IH, J = 15.0 Hz), 7.02 (d, IH, J = 8.7 Hz), 7.13 (dd, IH, J₁ = 3.6 Hz, J₂ = 5.1 Hz), 7.42 (d, IH, J= 3.3 Hz), 7.60-7.74 (m, 4H) and 10.1 (bs, IH); ¹³C NMR (DMSOd₆) δ 52.7, 58.5, 66.6, 118.1, 119.0, 119.1, 121.1, 128.3, 128.4, 131.1, 132.7, 135.0, 137.1, 139.8, 145.6 and 162.9; MS (ES) m/z 343 (M - H); Anal. (C₁₈H₂₀N₂O₃S) C₅ H₅ N. (E)-3-(Furan-2-vπ-N-(3-(hvdroxymethyl)-4-morpholinophenvπacrylamide (Compound 18b).

Compound 18b was synthesized, following the procedure used for preparation of 18a, by the reaction of 12b (0.500 g, 1.40 mmol) and DIBAL-H (7.02 mL, IM in dichloromethane) in dichloromethane (35 mL) and THF (5 mL) for 2 h at room temperature to furnish pure product (0.372 g, 81 %): mp 182 - 183 ⁰C; ¹H NMR (DMSO - d₆) δ 2.74-2.83 (m, 4H), 3.65-3.74 (m, 4H)₅ 4.54 (d, 2H, J = 5.1 Hz), 5.12 (t, IH, J = 5.4 Hz)₅ 6.58-6.67 (m, 2H), 6.82 (d, IH₅ J = 3.3 Hz), 7.02 (d, IH, J= 8.7 Hz), 7.35 (d, IH, J= 15.3 Hz), 7.61-7.71 (m, 4H)₅ 7.81 (d, IH₅ J= 1.5 Hz) and 10.1 (bs, IH); ¹³C NMR (DMSO - d₆) δ 53.1, 59.0, 67.1, 113.0, 114.8, 118.7, 119.4, 119.6, 120.1, 127.3, 135.4, 137.5, 145.5, 146.0, 151.4 and 163.5; MS (ES) m/z 327 (M - H); Anal. (C₁₈H₂₀N₂O₄) C₅ H₅ N. (E)-N-(3-Formyl-4-morpholinophenyl)-3-(thiophen-2-yl)acrylamide (Compound 19a)

To a solution of compound 18a (0.500 g, 0.44 mmol) in anhydrous THF at 0 ⁰C, pyridinium chlorochromate (0.188 g, 0.87 mmol) was added and stirred for 30 min and then at room temperature for 1 h. TLC analysis (1 :1 ΕtOAc/CHCl₃) revealed that the reaction was complete. The solvent was removed under vacuum and the crude product obtained was purified by column chromatography over Si gel using EtOAc/CHCl₃ (1 :4) as the eluent to afford pure 19a (0.107 g, 72 %): mp 218 - 219 °C; ¹H NMR (DMSO) δ 2.94-3.03 (m, 4H), 3.72-3.82 (m, 4H)₅ 6.53 (d, IH, J= 15.3 Hz), 7.11-7.17 (m, IH), 7.25 (d, IH, J= 8.7 Hz)₅ 7.45 (d, IH₅ J= 3.3 Hz), 7.66 (d, IH, J= 5.1 Hz), 7.74 (d, IH₅ J= 15.6 Hz)₅ 7.88 (dd, IH, J₁ = 2.4 Hz, J₂ = 8.7 Hz), 8.06 (d, IH, J = 2.7 Hz)₅ 10.2 (s, IH) and 10.3 (bs, IH). ¹³C NMR (CD₃COCD₃) δ 55.3, 67.4, 120.1, 121.0, 121.4, 126.8, 128.7, 129.1, 130.1, 131.7, 134.6, 136.1, 141.0, 152.5, 164.2 and 191.0. MS (ES) m/z 341 (M - H). Anal. (Ci₈Hi₈N₂O₃S) C, H, N. (E)-N-(3-Formyl-4-moφholinophenyl)-3-(fiiran-2-vπacrylamide (Compound 19b)

Compound 19b was synthesized, following the procedure used for preparation of 19a, by the reaction of 18b (0.188 g, 0.57 mmol) and pyridinium chlorochromate (0.247 g, 1.15 mmol) in anhydrous THF (10 mL) to furnish the pure product (0.115 g, 61 %): mp 231 - 232 ⁰C. ¹H NMR (DMSO) δ 2.94-3.03 (m, 4H), 3.73-3.82 (m, 4H), 6.53-6.65 (m, 2H), 6.86 (d, IH, J= 3.6 Hz), 7.24 (d, IH, J = 9.0 Hz), 7.38 (d, IH, J = 15.6 Hz), 7.82 (d, IH, J= 1.5 Hz), 7.89 (dd, IH, Ji = 2.5 Hz, J₂ = 8.8 Hz), 8.06 (d, IH, J = 2.7 Hz), 10.2 (s, IH) and 10.3 (bs, IH). ¹³C NMR (DMSO) δ 54.1, 66.2, 112.6, 114.8, 119.0, 119.1, 120.2, 126.0, 127.4, 128.2, 134.6, 145.3, 150.9, 151.2, 163.4 and 190.7. MS (ES) m/z 325 (M - H). Anal. (Ci₈Hi₈N₂O₄) C, H, N. 5-((E)-3-(Thiophen-2-yl)acrylamido)-2-morpholinobenzamide (Compound 20)

To a solution of compound 1 (0.050 g, 0.14 mmol) in anhydrous dichloromethane (4 mL) at room temperature a mixture of thionyl chloride (0.2 mL) and anhydrous DMF (0.2 mL) was added and the resulting mixture was stirred for 4 h at room temperature. The solvent was removed under reduced pressure. The resulting gum was dissolved in anhydrous CH₂Cl₂ (3 mL) and excess 50% aqueous ammonium hydroxide (3 mL) was added to achieve basic pH (-12) and stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the crude compound was purified by flash column chromatography (20 x 1 cm) on silica gel using EtOAc/CHCl₃ (1 :1 ) as eluent to furnish the pure amide 20 (0.013 g, 26 %): ¹H NMR (DMSO- d₆) δ 2.86-2.94 (m, 4H), 3.71-3.78 (m, 4H), 6.55 (d, IH, J= 15.3 Hz), 7.14 (dd, IH, J₁ = 3.6 Hz, J₂ = 5.1 Hz), 7.20 (d, IH, J = 8.7 Hz), 7.45 (d, IH, J = 3.0 Hz), 7.57 (bs, IH), 7.66 (d, IH, J =

5.1 Hz), 7.73 (d, IH, J = 15.6 Hz), 7.84 (dd, IH, J₁ = 2.7 Hz, _J2 = 8.7 Hz), 7.99 (d, IH, J = 2.7 Hz), 8.64 (bs, IH) and 10.2 (bs, IH). ¹³C NMR (DMSO-d₆) δ 52.9, 66.4, 120.4, 120.8, 121.1, 122.0, 128.5 (2C), 129.3, 131.3, 133.1, 135.1, 139.7, 146.2, 163.1 and 167.7; MS (ES) m/z 356 (M - H). Ethyl 5-((E)-3-(furan-2-yl)acrylamidoV2-moφholinobenzoate (Compound 43)

¹H-NMR (CDCl₃): δ 1.38 (t, 3H, J = 7.1 Hz ), 3.01 (t, 4H, J = 4.2 Hz), 3.84 (t, 4H, J =

4.2 Hz), 4.37 (q, 2H, J = 7.1 Hz), 6.38-6.47 (m, 2H), 6.58 (d, IH, J = 3.2 Hz), 7.03 (d, IH, J = 8.7 Hz), 7.45 (s, IH), 7.51 (d, IH, J = 15.1 Hz), 7.63(s, IH), 7.86 (m, 2H); ¹³C NMR (CDCl₃) δ

14.3, 53.0, 61.2, 67.1, 112.3, 114.6, 118.2, 119.8, 122.9, 124.4, 125.4, 129.0, 132.6, 144.3,

148.7, 151.2, 163.9, 167.3; MS (ES^") m/z 369 (M -H).

Benzyl 5-((E)-3-(raran-2-vl)acrvlamidoV2-morpholinobenzoate (44) ¹H-NMR (CDCl₃): δ 2.96 (t, 4H, J = 4.5 Hz), 3.71 (t, 4H, J = 4.5 Hz), 5.34 (s, 2H), 6.40 (d, IH, J = 15.3 Hz), 6.38-6.49 (m, IH), 6.61 (d, IH, J = 3.3 Hz), 7.05 (d, J = 8.7 Hz, IH), 7.32-7.44 (m, 4H), 7.45-7.7.56 (m, 4H), 7.81 (s, IH) 7.90 (bs, IH); MS (ES⁺) m/z 433 (M +H). Propyl 5-((EV3-(faran-2-yl)acrylamido)-2-morpholinobenzoate (Compound 45) 1H-NMR (CDCl₃): δ 0.99 (t, 3H, J = 7.4 Hz), 1.74 (sextet, 2H, J = 7.2 Hz ), 2.98 (bs,

4H), 3.82 (bs, 4H), 4.21 (t, 2H, J = 6.4 Hz), 6.35 - 6.60 (m, 3H), 6.98 (d, IH, J = 8.8 Hz), 7.39 (s, IH), 7.48 (d, IH, J =15.2 Hz), 7.86 (d, IH, J = 8.2 Hz), 7.95 (s, IH), 8.54 (s, IH); ¹³C NMR (CDCl₃) δ 11.0, 22.4, 53.4, 67.2, 67.5, 112.7, 114.8, 119.0, 120.1, 123.3, 124.8, 125.8, 129.1, 133.4, 144.7, 148.9, 151.6, 164.7, 168.0; MS (ES^') m/z 383 (M -H). Butyl 5-((E)-3-(faran-2-yl)acrylamido)-2-morpholinobenzoate (Compound 46)

¹H-NMR (CDCl₃): δ 0.94 (t, 3H, J = 7.5 Hz), 1.43 (sextet, 2H, J = 7.5 Hz), 1.70 (quintet, 2H, J = 7.5 Hz), 2.97 (bs, 4H), 3.82 (bs, 4H), 4.26 (t, 2H, J = 6.8 Hz), 6.39-6.58 (m, 3H), 6.98 (d, IH, J = 8.7 Hz), 7.28 (s, IH), 7.49 (d, IH, J = 15.3 Hz), 7.89 (m, 2H), 8.47 (bs, IH); ¹³C NMR (CDCl₃) δ 14.2, 19.6, 31.1, 53.4, 65.5, 67.5, 112.7, 114.7, 119.1, 120.1, 123.3, 124.9, 125.9, 129.0, 133.5, 144.6, 148.9, 151.6, 164.8, 168.0; MS (ES^') m/z 397 (M-H). Methyl 5-(cinnamamido)-2-moφholinobenzoate (Compound 52)

¹H-NMR (CDCl₃): δ 3.03 (t, 4H, J = 4.5Hz), 3.86 (t, 4H, J = 4.5Hz), 3.89 (s, 3H), 6.52 (d, IH, J = 15.3 Hz), 7.06 (d, IH, J= 8.7Hz), 7.35-7.41 (m, 4H), 7.52-7.56 (m, 2H), 7.76(d, IH, J = 15.3Hz), 7.93 (bs, IH); ¹³C NMR (CDCl₃) δ 52.2, 53.0, 67.1, 119.7, 121.0, 123.5, 124.5, 124.9, 127.9, 128.8,129.9, 132.8, 134.6, 142.1, 148.9, 164.7, 167.6.

Methyl 5-((E)-3-(pyridin-3-yl)aciΥlamido)-2-morpholinobenzoate (Compound 55)

¹H-NMR (CDCl₃): δ 3.00 (t, 4H, J = 4.5 Hz ), 3.84 (t, 4H, J = 4.5 Hz), 3.87 (s, 3H), 6.69 (d, IH, J = 15.6 Hz ), 7.03 (d, IH, J = 8.7 Hz), 7.29-7.38 (m, IH), 7.67-7.79 (m, 2H), 7.86 (d, IH, J = 7.2 Hz ), 8.00 (s, IH), 8.44 (s, IH), 8.59 (s, IH), 8.77 (s, IH); ¹³C NMR (CDCl₃) δ 52.2, 53.0, 67.1, 119.8, 123.1, 123.4, 123.9, 124.8, 130.7, 132.4, 134.9, 138.5, 148.9, 149.1, 150.4, 163.3, 167.5; MS (ES⁺) m/z 368 (M+H). Methyl 5-((E)-3-(pyridin-4-yl)acrylamido)-2-morpholinobenzoate (Compound 51)

¹H-NMR (DMSOd₆): δ 2.91 (t, 4H, J = 4.5 Hz), 3.69 (t, 4H, J = 4.5Hz), 3.82 (s, 3H), 7.04 (d, IH, J = 15.9 Hz), 7.13 (d, IH, J = 9 Hz), 7.57 (d, IH, J = 15.9 Hz), 7.65 (d, IH, J = 5.1 Hz), 7.80 (dd, IH, J₁ = 2.4 Hz, J₂ = 8.7 Hz), 8.02 (d, IH, J = 2.7 Hz), 8.31 (s, IH), 8.68 (d, 2H, J = 4.2 Hz) ; ¹³C NMR (DMSOd₆) δ 52.5, 53.0, 66.8, 79.6, 120.2, 122.1, 122.6, 124.0, 125.0, 127.8, 133.6, 137.6, 143.6, 148.1, 149.8, 163.0, 167.9; MS (ES⁺) m/z 368 (M⁺H). 5-(Cinnamamido)-2-morpholinobenzoic acid (53) ¹H-NMR (DMSO-d₆): δ 3.05 (t, 4H, J = 4.2 Hz), 3.80 (t, 4H, J = 4.2 Hz), 6.82 (d, IH, J = 15.9 Hz), 7.38-7.51 (m, 3H), 7.57-7.72 (m, 4H), 8.03 (dd, IH, J₁ = 2.4Hz, J2, = 8.7 Hz), 8.31 (d, IH, J = 2.7 Hz ), 10.50 (s, IH).

5-((E)-3-(Pyridin-3-vDacrylamido)-2-morpholinobenzoic acid (Compound 56) 1H-NMR (DMSOd₆): δ 3.09 (t, 4H, J = 4.5 Hz ), 3.81 (t, 4H, J = 4.5 Hz), 7.14 (d, IH, J

= 15.9 Hz), 7.71 (d, IH, J = 2.7 Hz), 7.75 (d, IH, J = 9.9 Hz ), 7.91 (dd, IH, J₁ = 5.4 Hz, J₂ = 8.1Hz), 8.05 (dd, IH, J₁ = 2.7 Hz, J₂ = 9.0 Hz), 8.37 (d, IH, J = 2.4 Hz ), 8.53 (d, IH, J = 7.0 Hz ), 8.81 (dd, IH, J₁ = 1.5 Hz, J₂ = 5.4 Hz), 9.07 (d, IH, J = 1.8 Hz ), 10.91 (s, IH; ¹³C NMR (DMSO-d₆) δ 53.3, 66.8, 121.7, 124.2, 124.7, 125.8, 126.7, 127.0, 133.3, 135.6, 138.0, 140.2, 144.8, 145.5, 145.7, 163.3, 166.9; MS (ES⁺) m/z 354 (M+H).

5-((E)-3-(Pyridin-4-yDaciylamido)-2-morpholinoberizoic acid (Compound 54)

¹H-NMR (DMSOd₆): δ 3.06 (bs , 4H), 3.80 (bs, 4H), 7.28 (d, IH, J = 16.2 Hz), 7.65- 7.76 (m, 2H), 8.05 (bs, 3H), 8.34 (s, IH), 8.87 (d, 2H, J = 5.4 Hz), 10.94 (s, IH). (E)-I -(4-(4-nitrophenoxy)phenyl)-3-(dimethylamino)prop-2-en-l -one (Compound 57) 1H-NMR (CDCl₃): δ 2.96 (s, 3H), 3.17 (s, 3H), 5.70 (d, IH, J = 12.3 Hz), 7.05-7.13 (m,

4H), 7.84 (d, IH, J = 12.3Hz), 7.98 (d, 2H, J = 8.7 Hz), 8.22 (d, 2H, J = 9.3 Hz; ¹³C NMR (CDCl₃) δ 38.0, 46.0, 117.9 (2C), 120.4, 126.6 (2C), 130.3, 137.9, 143.0, 155.1, 157.2, 163.3; MS (ES⁺) m/z 313 (M+H).

Table 1

Compd. Structure No. IC₅₀ (μM)^fl

NH, _>N'^Nγ^NH_

NH, 275 ± 11.8

H₂N^ΛN'^N<

" IC₅₀Valueswere determined by fluorescence resonance energy transfer (FRET) assay Measurements were carried out in triplicate and average value with standard deviation is reported. Table 2: S. aureus SrtAa₅9 inhibition of the synthesized compounds.

aIC₅₀Valueswere determined by fluorescence resonance energy transfer (FRET) assay. Measurements were carried out in triplicate and average value with standard deviation is reported.

Claims

CLAIMSWhat is claimed:

1. A method of inhibiting a bacterial sortase in a subject, the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NRi, where Ri is H or a substituted or unsubstituted alkyl;

E is selected from the group consisting of: CH₂, O and N;

L₁ and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and L is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

I is linking group and is a C₂ to C₅ alkenyl containing a single carbon- carbon double bond or C₂ to C₅ alkynyl containing a single carbon-carbon triple bond;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORi₃, COORi₄, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclyl alkyl ; R₆ through Rg are each independently selected from the group consisting of: H, OH, and substituted or unsubstituted alkyl;

R₁₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR₁₅R₁₆;

R_H is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl; and

R₁₅ and R₁₆ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

2. A method of inhibiting a bacterial sortase in a subject, the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NR₁, where Ri is H or a substituted or unsubstituted alkyl;

E is a heteroatom selected from the group consisting of: CH₂, O and N;

F is a single, optional N heteroatom; when present, F may be in a para, meta or ortho position with respect to the point of attachment; Li and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and L is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl; I is linking group and is a C₂ to C₅ alkenyl containing a single carbon- carbon double bond or C₂ to C₅ alkynyl containing a single carbon-carbon triple bond;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORi₄, COORi₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl ;

R₉ through Rn are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl;, provided that, when F is present at a given position, the respective R₉-R₁₃ substituent group will be absent

Ri₄ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₆Ri₇;

Ri₅ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl; and

Ri₆ and Ri₇ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

3. A method of inhibiting a bacterial sortase in a subject, the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

Li is an optional linking groups, where 1 is selected from 0 or 1 and L is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

R₂ through R₆ are each independently selected from the group consisting of: H, OH; CORi₀, COOR_n, NRi₄Ri₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

R₇ through R₉ are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl;

Rio is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR]₂R)₃;

Ri₂ and Rj₃ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl; and R_H and R₁₅ are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl and substituted or unsubstituted aralkyl, or a pharmaceutically acceptable salt thereof.

4. A method of inhibiting a bacterial sortase in a subject, the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

R₁ through R₉ are each independently selected from the group consisting of: H, OH, NR]₆R₁₇, CORi₂, COORi₃, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

Rio and Rn are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

Ri₂ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₄Ri_S;

R₁₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl; and

R_H through R₁₇ are each independently selected from the group consisting of: H, O and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

5. A method of inhibiting a bacterial sortase in a subject, the method comprising the steps of administering to the subject a compound having a structure selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

6. The method of any one of claims 1-5 where the sortase enzyme is SrtA, SrtB, SrtC, SrtD or a homolog of any of the foregoing.

7. The method of claim 6 where the sortase enzyme is SrtA.

8. The method of any one of claims 1-5 where said inhibition reduces, at least in part, the ability of a bacterium to infect a cell of the subject or reduces, at least in part, virulence of a bacterium towards the subject.

9. The method of claim 8 where said bacterium is a Gram-positive bacterium.

10. The method of claim 8 where said bacterium is Staphylococcus aureus.

11. The method of claim 8 where said bacterium is a Gram-negative bacterium.

12. The method of claim 8 where the bacterium is resistant to one or more antibiotics.

13. The method of claim 8 where the bacterium is a methicillin-resistant Staphylococcus aureus, a vancomycin-resistant enterococcus, a vancomycin-intermediate Staphylococcus aureus or a vancomycin-resistant Staphylococcus aureus.

14. The method of any one of claims 1-5 where said inhibition reduces, at least in part, a bacterial infection.

15. The method of any one of claims 1 -5 where said inhibition reduces, at least in part, a disease state or condition caused by a bacterial infection.

16. The method of claim 15 where the disease state or condition is selected from the group consisting of: endocarditis, osteomyelitis, septic arthritis pneumonia, toxic shock syndrome, skin infections, meningitis, anthrax, B. cereus food poisoning, tetanus, clostridial myonecrosis, pneumonia, toxic shock syndrome, skin infection, meningitis, ear infection, strep throat, rheumatic fever, scarlet fever, puerperal fever, listeriosis, perinatal septicemia, encephalitis, intrauterine infection, sinusitis, severe invasive infection, and a combination of the foregoing.

17. The method of claims 15 where the disease state or condition is endocarditis.

18. The method of any one of claims 1 -5 further comprising administering a secondary antibiotic.

19. A method of reducing bacterial infection or bacterial virulence in a subject the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

E is selected from the group consisting of: CH₂, O and N;

Li and L₂ are optional linking groups, where 1 and 2 are independently selected from 0 or 1 and L is each independently selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORi₃, COORi₄, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

R₆ through R₈ are each independently selected from the group consisting of: H, OH, and substituted or unsubstituted alkyl; Ri₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR₁₅Ri₆;

Ri ₅ and Ri₆ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

20. A method of reducing bacterial infection or bacterial virulence in a subject bacterial virulence in a subject the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NRj, where Ri is H or a substituted or unsubstituted alkyl;

E is a heteroatom selected from the group consisting of: CH₂, O and N;

F is a single, optional N heteroatom; when present, F may be in a para, meta or ortho position with respect to the point of attachment;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, CORi₄, COORi ₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Cj to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclyl alkyl;

R₉ through Rn are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl, provided that, when F is present at a given position, the respective R₉-R₁₃ substituent group will be absent;

Ri₄ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRj₆Ri₇;

Ri₆ and Rj₇ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

21. A method of reducing bacterial infection or bacterial virulence in a subject the method comprising the steps of administering to the subject a compound having the structure:

wherein:

A is selected from the group consisting of: S, CH₂, O and N; B is selected from the group consisting of: S, O and NRi, where R₁ is H or a substituted or unsubstituted alkyl; Li is an optional linking groups, where 1 is selected from 0 or 1 and L is selected from substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

R₂ through R₆ are each independently selected from the group consisting of: H, OH; CORi₀, COORn, NRi₄R₁₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl;

Rio is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₂Ri₃;

Ri₂ and Ri₃ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl; and

Ri₄ and Rj₅ are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl and substituted or unsubstituted aralkyl, or a pharmaceutically acceptable salt thereof.

22. A method of reducing bacterial infection or bacterial virulence in a subject, the method comprising the steps of administering to the subject a compound having the structure:

where:

A is selected from the group consisting of: S, CH₂, O and N; I is linking group and is a C₂ to C₅ alkenyl containing a single carbon- carbon double bond or C₂ to C₅ alkynyl containing a single carbon-carbon triple bond;

R₁ through R₉ are each independently selected from the group consisting of: H, OH, NR₁₆Rj₇, COR₁₂, COORi₃, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl ;

Ri₀ and Rn are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl and substituted or unsubstituted alkynyl;

Ri₂ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₄Ri₅;

Ri₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl; and

Ri₄ and Ri₇ are each independently selected from the group consisting of: H, O and substituted or unsubstituted alkyl, or a pharmaceutically acceptable salt thereof.

23. A method of reducing bacterial virulence in a subject the method comprising the steps of administering to the subject a compound a structure selected from the group consisting of:

or a pharmaceutically acceptable salt thereof 24. The method of any one of claims 19-23 where the administration decreases, in whole or part, the activity of a bacterial sortase enzyme.

25. The method of claim 24 where the bacterial sortase enzyme is SrtA, SrtB, SrtC, SrtD or a homolog of any of the foregoing.

26. The method of claim 24 where the bacterial sortase enzyme is SrtA.

27. The method of claim 24 where said decrease in activity reduces, at least in part, the ability of a bacterium to infect the cells of the subject.

28. The method of any one of claims 19-23 where the bacterial infection or virulence is caused, at least in part, by a Gram-positive bacterium.

29. The method of claim 28 where said Gram-positive bacterium is Staphylococcus aureus.

30. The method of any one of claims 19-23 where the bacterial infection or virulence is caused, at least in part, by a Gram-negative bacterium.

31. The method of any one of claims 19-23 where the bacterial infection or virulence is caused, at least in part, by an antibiotic resistant bacterium.

32. The method of claim 31 where the antibiotic resistant bacterium is a methicillin-resistant Staphylococcus aureus, a vancomycin-resistant enterococcus, a vancomycin-intermediate Staphylococcus aureus or a vancomycin-resistant Staphylococcus aureus.

33. The method of any one of claims 19-23 further comprising administering a secondary antibiotic.

34. A compound of the formula Ia:

(Ia)

wherein:

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NRi, wherein R] is H or a substituted or unsubstituted alkyl; E is selected from the group consisting of: CH₂, O and N; Li and L₂ are optional linking groups, wherein 1 and 2 are independently selected from 0 and 1, and L is independently selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl; I is a linking group, selected from the group consisting of a C₂ to C₅ alkenyl containing a single carbon-carbon double bond and a C₂ to C₅ alkynyl containing a single carbon-carbon triple bond;

R₂ through R₅ are each independently selected from the group consisting of: H, OH,

COR₁₃, COORi₄, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, C₁ to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl, wherein R₁₃ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NR₁₅Ri₆; wherein R₁₄ is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, and substituted or unsubstituted aralkyl; wherein R₁₅ and R₁₆ are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl; and R₆ through R₈ are each independently selected from the group consisting of: H, OH, and substituted or unsubstituted alkyl; provided that whenever A is S, R₂ cannot be COOH when B is NH, E is O, I is a C₂ alkenyl in trans configuration, Li and L₂ are both O, and R₃ through R₈ are H; or a pharmaceutically acceptable salt thereof.

35. A compound according to claim 34, wherein R₂ is COOCH₃, or a pharmaceutically acceptable salt thereof.

36. A compound according to claim 35, wherein A is S, or a pharmaceutically acceptable salt thereof.

37. A compound according to claim 36, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both O, or a pharmaceutically acceptable salt thereof.

38. A compound according to claim 36, wherein: B is NH, E is O, I is a C₂ alkynyl, R₃ through R₈ are H, and 1 and 2 of L₁ and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

39. A compound according to claim 36, wherein: B is NH, E is O, I is a C₂ alkenyl in the cis configuration, R₃ through R₈ are H, and 1 and 2 of Lj and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

40. A compound according to claim 36, wherein: B is NH, E is O, I is a C₃ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

41. A compound according to claim 36, wherein: B is NH, E is CH₂, 1 is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

42. A compound according to claim 36, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, 1 of Li is 0, 2 of L₂ is 1 and L in L₂ is CH₂, or a pharmaceutically acceptable salt thereof.

43. A compound according to claim 35, wherein A is O, or a pharmaceutically acceptable salt thereof.

44. A compound according to claim 43, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Lj and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

45. A compound according to claim 43, wherein: B is NH, E is CH₂, 1 is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

46. A compound according to claim 34, wherein R₂ is COOH, or a pharmaceutically acceptable salt thereof.

47. A compound according to claim 46, wherein A is S, or a pharmaceutically acceptable salt thereof.

48. A compound according to claim 47, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Lj and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

49. A compound according to claim 47, wherein: B is NH, E is O, I is a C₂ alkenyl in the cis configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

50. A compound according to claim 47, wherein: B is NH, E is CH₂, 1 is a C₂ alkenyl in the trans configuration, R₃ through Rg are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

51. A compound according to claim 47, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, 1 Of L₁ is 0, 2 of L₂ is 1 and L in L₂ is CH₂, or a pharmaceutically acceptable salt thereof.

52. A compound according to claim 46, wherein A is O, or a pharmaceutically acceptable salt thereof.

53. A compound according to claim 52, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

54. A compound according to claim 34, wherein: R₂ is C(=O)H, or a pharmaceutically acceptable salt thereof.

55. A compound according to claim 54, wherein: A is S, or a pharmaceutically acceptable salt thereof.

56. A compound according to claim 55, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

57. A compound according to claim 54, wherein: A is O, or a pharmaceutically acceptable salt thereof.

58. A compound according to claim 57, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

59. A compound according to claim 34, wherein: R₂ is CH₂OH, or a pharmaceutically acceptable salt thereof.

60. A compound according to claim 59, wherein: A is S, or a pharmaceutically acceptable salt thereof.

61. A compound according to claim 60, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

62. A compound according to claim 59, wherein: A is O, or a pharmaceutically acceptable salt thereof.

63. A compound according to claim 62, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

64. A compound according to claim 34, wherein: R₂ is C(=0)NH₂, or a pharmaceutically acceptable salt thereof.

65. A compound according to claim 64, wherein: A is S, or a pharmaceutically acceptable salt thereof.

66. A compound according to claim 65, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 are both 0, or a pharmaceutically acceptable salt thereof.

67. A compound according to claim 34, wherein: R₂ is C(=O)-ORi₄, wherein Ri₄ is selected from a group consisting Of C₂H₅, C₃H₇, C₄Hg, and C₇H₇, or a pharmaceutically acceptable salt thereof.

68. A compound according to claim 67, wherein: A is O, or a pharmaceutically acceptable salt thereof.

69. A compound according to claim 68, wherein: B is NH, E is O, I is a C₂ alkenyl in the trans configuration, R₃ through R₈ are H, and 1 and 2 of Li and L₂ are both 0, or a pharmaceutically acceptable salt thereof.

70. A compound according to claim 69, wherein: R]₄ is C₂H₅, or a pharmaceutically acceptable salt thereof.

71. A compound according to claim 69, wherein: Ri₄ is CH₂-C₆Hs, or a pharmaceutically acceptable salt thereof.

72. A compound according to claim 69, wherein: Ri₄ is C₃H₇, or a pharmaceutically acceptable salt thereof.

73. A compound according to claim 69, wherein: Rj₄ is C₄H₉, or a pharmaceutically acceptable salt thereof.

74. A compound of the formula Ib:

(Ib)

wherein:

A is selected from the group consisting of:, S, CH₂, O and N;

B is selected from the group consisting of: S, O and NR₁, wherein Ri is H or a substituted or unsubstituted alkyl;

E is selected from the group consisting of: CH₂, O and N;

L] and L₂ are optional linking groups, wherein 1 and 2 are independently selected from 0 and 1, and L is independently selected from the group consisting of: substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, and substituted or unsubstituted alkynyl;

I is a linking group, selected from the group consisting of a C₂ to C₅ alkenyl containing a single carbon-carbon double bond and a C₂ to C₅ alkynyl containing a single carbon-carbon triple bond;

R₂ through R₅ are each independently selected from the group consisting of: H, OH, COR]₄, COORi₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl, wherein Ri₄ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRj₆Ri₇; wherein Ru is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, and substituted or unsubstituted aralkyl; wherein Rj₆ and Rp are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl; and

R₉ through Ri₃ are each independently selected from the group consisting of: H, OH, and substituted or unsubstituted alkyl, provided that when F is present at a given position, the respective R₉ through Rj₃ substituent group will be absent; or a pharmaceutically acceptable salt thereof.

75. A compound according to claim 74, wherein: B is NH, E is O, and I is a C₂ alkenyl in the trans configuration, or a pharmaceutically acceptable salt thereof.

76. A compound as in claim 75, wherein R₂ is COOH, or a pharmaceutically acceptable salt thereof.

77. A compound according to claim 76, wherein: F is absent, 1 and 2 of Li and L₂ are both O,

R₃ through R₅ and R₉ through Ri₃ are H, or a pharmaceutically acceptable salt thereof.

78. A compound according to claim 76, wherein: F is present in the meta position, 1 and 2 of Li and L₂ are both 0, Ri₀ is absent, R₃ through R₅, R₉ and Rn through R₁₃ are H, or a pharmaceutically acceptable salt thereof.

79. A compound according to claim 76, wherein: F is present in the para position, 1 and 2 of Li and L₂ are both 0, Rn is absent, R₃ through R₅, R₉ through Ri₀ and Ri₂ through Ri₃ are H, or a pharmaceutically acceptable salt thereof.

80. A compound as in claim 75, wherein R₂ is COOCH₃, or a pharmaceutically acceptable salt thereof.

81. A compound according to claim 80, wherein: F is absent, 1 and 2 of Li and L₂ are both 0,

R₃ through R₅ and R₉ through Rj₃ are H, or a pharmaceutically acceptable salt thereof.

82. A compound according to claim 80, wherein: F is present in the meta position, 1 and 2 of Li and L₂ are both 0, Ri₀ is absent, R₃ through R5, R9 and Rn through R_]3 are H, or a pharmaceutically acceptable salt thereof.

83. A compound according to claim 80, wherein: F is present in the para position,l and 2 of Li and L₂ are both 0, Rn is absent, R₃ through R₅, R₉ through Rj₀ and R₎₂ through Rj₃ are H, or a pharmaceutically acceptable salt thereof.

84. A compound of the formula Ic: (Ic)

Wherein

A is selected from the group consisting of: S, CH₂, O and N;

B is selected from the group consisting of: S, O and NRi, wherein Rj is H or a substituted or unsubstituted alkyl;

Li is an optional linking groups, where 1 is selected from 0 or 1 and L is selected from the group consisting of substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl;

I is a linking group, s^*eje^j^|frpm the group consisting of a C₂ to C₅ alkenyl containing a single carbon-carbon double bond in either the cis or trans configuration and a C₂ to C₅ alkynyl containing a single carbon-carbon triple bond; and

R₂ through R₆ are each independently selected from the group consisting of: H, OH, CORio, COORi i, NR)₄Ri₅, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, Ci to C₇ alcohols, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aralkyl and substituted or unsubstituted heterocyclylalkyl, wherein R^ is selected from the group consisting of: H, substituted or unsubstituted alkyl, and NRi₂Ri₃, wherein Rn is selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heterocyclylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl and substituted or unsubstituted aralkyl, wherein R]₂ and Rn are each independently selected from the group consisting of: H and substituted or unsubstituted alkyl, and wherein Ri₄ and R)₅ are each independently selected from the group consisting of: H, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl and substituted or unsubstituted aralkyl; and

R₇ through R9 are each independently selected from the group consisting of: H, OH and substituted or unsubstituted alkyl; or a pharmaceutically acceptable salt thereof.

85. A compound as in claim 84, wherein R₃ is COOCH₃, or a pharmaceutically acceptable salt thereof.

86. A compound as in claim 85, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is NH-CH₂-C₆H₅, R₄ through R₉ are H, and 1 of Li is 0, or a pharmaceutically acceptable salt thereof.

87. A compound as in claim 85, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is NH-C₆H₅, R₄ through R₉ are H, and 1 of Li is 0, or a pharmaceutically acceptable salt thereof.

88. A compound as in claim 85, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is N-(CH₂-C₆H₅)₂, R₄ through R₉ are H, and 1 of Li is 0, or a pharmaceutically acceptable salt thereof.

89. A compound as in claim 84, wherein: R₃ is COOH, or a pharmaceutically acceptable salt thereof.

90. A compound as in claim 89, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is NH-C₆H₅, R₄ through R₉ are H, and 1 of Lj is 0, or a pharmaceutically acceptable salt thereof.

91. A compound as in claim 89, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is N-(CH₂-C₆Hs)₂, R₄ through R₉ are H, and 1 of Li is 0, or a pharmaceutically acceptable salt thereof.

92. A compound as in claim 89, wherein: A is S, B is NH, I is a C₂ alkenyl in the trans configuration, R₂ is N-(C₂H₄OH)₂, R₄ through R₉ are H, and 1 of Li is 0, or a pharmaceutically acceptable salt thereof.