US 20070134729A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0134729 A1 Christensen et al. (43) Pub. Date: Jun. 14, 2007

(54) NOVELANTIBACTERIAL AGENTS Publication Classification (51) Int. Cl. (76) Inventors: Burton G. Christensen, Alamo, CA A6II 38/16 (2006.01) (US); Edmund J. Moran, San A6II 3L/43 (2006.01) Francisco, CA (US); John H. Griffin, A6II 3/545 (2006.01) Atherton, CA (US); J. Kevin Judice, El A6II 3 L/407 (2006.01) Granada, CA (US); YongOi Mu, Los (52) U.S. Cl...... 435/7.1: 514/8: 514/192: 514/200; Altos, CA (US); John L. Pace, San 514/210.09: 514/210.15 Anselmo, CA (US); Mathai Mammen, (57) ABSTRACT Redwood City, CA (US); James Aggen, This invention relates to novel multibinding compounds San Francisco, CA (US) (agents) that are antibacterial agents. The multibinding com pounds of the invention comprise from 2-10 ligands Correspondence Address: covalently connected by a linker or linkers, wherein each of said ligands in their monovalent (i.e., unlinked) state have THERAVANCE, INC. the ability to bind to a an enzyme involved in 901 GATEWAY BOULEVARD biosynthesis and metabolism, a precursor used in the Syn SOUTH SAN FRANCISCO, CA 94080 (US) thesis of the bacterial cell wall and/or the bacterial cell surface thereby interfere with the synthesis and/or metabo lism of the cell wall. In particular the multibinding com (21) Appl. No.: 11/603,287 pounds of the invention comprise from 2-10 ligands (22) Filed: Nov. 21, 2006 covalently connected by a linker or linkers, wherein each of said ligands has a ligand domain capable of binding to Related U.S. Application Data binding proteins, a transpeptidase enzyme, a Sub strate of a transpeptidase enzyme, a beta-lactamase enzyme, (63) Continuation of application No. 09/457,926, filed on pencillinase enzyme, cephalosporinase enzyme, a transgly Dec. 8, 1999, which is a continuation-in-part of coslase enzyme, or a transglycosylase enzyme Substrate; application No. 09/317,198, filed on May 24, 1999, Preferably, the ligands are selected from the beta lactam or now abandoned. glycopeptide class of antibacterial agents. Patent Application Publication Jun. 14, 2007 Sheet 1 of 25 US 2007/0134729 A1 ......

O-o-o-o O-o--N-o-O O s W R On 101 nu-0 W Of O Of 691 OH Patent Application Publication Jun. 14, 2007 Sheet 2 of 25 US 2007/0134729 A1

9

Owl, Owl, o, -o ?oNu-1a1nu-N o, Patent Application Publication Jun. 14, 2007 Sheet 3 of 25 US 2007/0134729 A1

". Patent Application Publication Jun. 14, 2007 Sheet 4 of 25 US 2007/0134729 A1

O S CH R-C-NH-CH-cfcC.B I A CH3 O=C-N-CH-COOH R WOWPROPRIETARY SIDE CHAIN NAAME

(O)-CH2 - Penicillin G (O-Octe - . Penicillin V C- OCH3 R Oxacilin - (R = R2 = H) Cloxacilin R2 "Yo-Yo, (R1 = Cl; R2 = H) (O) (R1 = R2 = Cl) {O) Nofcillin

R -O-H- NH2 (R = H) (R = OH)

CH- loor (R = H) Corbenicillin indanyl (R = 5-indanol) S H COOH FIG. 5 Patent Application Publication Jun. 14, 2007 Sheet 5 of 25 US 2007/0134729 A1

(O) ch pa

(r Piperacilin O

FIGURE5 CONTD Patent Application Publication Jun. 14, 2007 Sheet 6 of 25 US 2007/0134729 A1

FIG. 6A R.--NH 7

p COO COMPOUNO Cephen nucleus (IRADE NAMES) R R2 First-Generation - O (KEFLIN)Cephalothin SyCHl, -CH20cc 3. W= W-W (WGF,Cefozolin KEFZOL, others) TaiA" - - - wo CH2S-l Sct,

(KEFLEI,Cephalexin KEFLEX) (O)- CH- -CH3 NH2 (DURICEF,Cefoodroxid ULTRACEF) HO -O-GH - -CH3 NH2 Second-Generation S. (O) CH- - CH2S y (MANDOL ) OH CH O (MEFOXIN) f Srych, -- -CH2OC 2 NH2 CH- - Cl (CECLOR) NH2 O - O (KEF UROX, ZINACEF) -CH2OC Cefuroxime oxetil t N OCH NH2 (CEFTIN) 3. loracorbef #-F. CH- -Cl (CEFZIL) NH2 N-W (MONOCID) CH- --- Cl2S 1 w-N OH CHSO3 ? N-N (CEFOIAN)Cefoteton H2NCSY -CH2S1l, N Patent Application Publication Jun. 14, 2007 Sheet 7 of 25 US 2007/0134729 A1

1 R-(-NHJi2–N4 R2 COO nucleus COMPOUND (TRADE NAMES) R R Second-Generotion CONTINUED N-W (PRECEF) C-cle - Ches? N N CH2NH2 CH2COOH

Third-Generation W O Cefotoxime H wis D N -CH2OC < O (CLAFORAN) 2 SOCH CH

proxetilCefoodoxime H wsW N a -CH2OCH3 (VANTIN) 2 OCH

W HW 1s | N -H (CEFIZOX) 2 NO CH3

Ceftrioxone HW 1sW D N H3C :w-Ns-OH C (ROCEPHIN) 2 SOCH -CH2S1v-so N-N Cefoperozone HO -O- CH- l nk (CEFOBID) -CH2S NHCO CH

oryr") C2H5 FIG. 6B-1 Patent Application Publication Jun. 14, 2007 Sheet 8 of 25 US 2007/0134729 A1

COMPOUND (IRADE NAMES). R, R2 . Third-Generotion (continued) Ceftazidinein - ... Ji w O) (FORTAZ, others) HNS N SOC(CH )2COOH -CHN O - F.ourth-Generation

- 3. Cefepine H2 W lS1. N. -CH2N K OCH3. + Cefoxitin, a , has a -OCH3 group at position 7 of cephem nucleus. is the acetyloxyethyl ester of cefuroxime. ++ Loracarbe?, a carbocephem, has a carbon instead of sulfur at position 1 of cephem nucleus. # Cefoodoxime proxeul has a -COOCH(CH3)OCOOCH(CH3)2 group at position 4 of cephen nucleus,

FIG.6B-2 Patent Application Publication Jun. 14, 2007 Sheet 9 of 25 US 2007/0134729 A1

R3 WH S 5. Ah W. R Cephern O "S-4 -LINKERHR1C" O Cephen p O HO O p O OH YO

4. S K LINKER R3 -R his Cephem R 'i. O -LINKERH-R- O O $2.A.-o Cephem P HO- O O OH

FIG 7A Patent Application Publication Jun. 14, 2007 Sheet 10 of 25 US 2007/0134729 A1

CH3 O Carbopenem

Rik his # h is Cephen O 2 LINKERH -R N-Wall? . A O OH O f OH Penen Ho H H S H3C -LINKER-R-N-Wu: 4. Carbopenem OH Cephern O ^OH

H 'S O "y-r-W-1 Cephem Penen O O OH

H S. 'Y-R-LINKERHW RSW3H H S N O N N-NA R4 Penen HO O O O Cephern O O OH

FIG 7B Patent Application Publication Jun. 14, 2007 Sheet 11 of 25 US 2007/0134729 A1

gh

Carbopenem O O Carbopenem

R9

Penen HO O O Carbopenem

HO R 9 H3C R10. LINKER f R.'-SJ... Carbopenem O OH O f- OH Penon O O

S S H H SOH , W-Ré-R-LINER) LINKER (, -a, Penen OH HO O Penen O O

:Ss: LINKER-R "...itH H S Aenen OH O f- OH Penon O O SJf & -R-LINKER "),it is A m O Penam O O O f OH Penan O FIG. 8A Patent Application Publication Jun. 14, 2007 Sheet 12 of 25 US 2007/0134729 A1

Likewise: Likewise: Oxocephem-Cephem Irinem-Cephem Oxocephem-Cepham Irinem-Cepham -Penan Irinem-Penam Oxocephem-Penen Irinen-Penen Oxocephem-Carbopenem Irinem-Carbopenen Oxacephen-Carbocephem Irinem-Carbocephem Oxacephern-Irinem Trinern-Irinen : Oxacephen- Irinen-Monoboctorn Oxacephem-Monobactam" Irinem-Monobactam"

Likewise. Likewise: - Cephem Monobactorm-Cephem Carbocephem - Cepharm' Monobactam-Cepham Carbacephem -Penam Monobactorm-Penam Carbacephem -Renem Monobactam-Penen Carbocephem -Carbopenem Monobactom-Carbopenern Carbacephem -Carbocephem Monobactorn-Monoboctorn Corbacephem - Irinem Monobactam-Monobactam" Carbocephen -Monobactam Monobactam-Monobactam" Carbacephem -Monobactam" FIG. 8B Patent Application Publication Jun. 14, 2007 Sheet 13 of 25 US 2007/0134729 A1

R3 R h S O Cephem O "y-r-LIKERN WHIMe Glycopeptide O ^OH H2N () H O 3. H t S RW-y R4-LINKER R-NH OH G/ycopeptide fid Cephem O N N1 (Vancomycin) O ^OH NHMe O S H. H. Me Cephem i N R 3. -LINKERINKER R-N OH (Vancomycin)Glycopep tide NH2 - HO-Yo O

O H. H.g S Glycopeptide H R3-N P Cephem' Rio's LINKER1' 1 y)-N-1 epnem MeFHW O or OH O Me H. H. Glycopeptide R 3. S Cephem A

O O OH

O - p 3 H. H. S o Me??W .'R NLINKER R g"), -R? Glycopeptide O Cephem' (Vancomycin) NH2 O OH FIG. 9 Patent Application Publication Jun. 14, 2007 Sheet 14 of 25 US 2007/0134729 A1

O R-LINKER10 R NH2 ycopeptidetid MeFHW ( Voncornycinyc ) OH

O W-R6 LINKER R-WH 0 (Vanconycin)Glycopeptide Penen H NHMe

S. H R Me O va N-R LINKER-R-W Glycopeptide W O OH (Vancomycin) Penon HO O O NH2

Likewise Glycopeptide linked to these betalactam : Oxacephem- Irinem- Glycopeptide Antibiotic Carbacephen-Glycopeptide Antibiotic Monobactam- Glycopeptide Antibiotic Monobactam- Glycopeptide Antibiotic

FIG 10 Patent Application Publication Jun. 14, 2007 Sheet 15 of 25

-__^30/y-HN#30//--HN Patent Application Publication Jun. 14, 2007 Sheet 16 of 25

+?AN 9

W O SH·WH Patent Application Publication Jun. 14, 2007 Sheet 17 of 25 US 2007/0134729 A1 Patent Application Publication Jun. 14, 2007 Sheet 18 of 25 US 2007/0134729 A1 HHN-KO NLaHSS+õHNO Erso“T), •/O ·HS

Patent Application Publication Jun. 14, 2007 Sheet 20 of 25 US 2007/0134729 A1

(S 5 HOOO

S.

n n Patent Application Publication Jun. 14, 2007 Sheet 21 of 25 US 2007/0134729 A1

e HOOO!O•======2 N Patent Application Publication Jun. 14, 2007 Sheet 22 of 25 US 2007/0134729 A1 HŽojO Patent Application Publication Jun. 14, 2007 Sheet 23 of 25 US 2007/0134729 A1 Patent Application Publication Jun. 14, 2007 Sheet 24 of 25 US 2007/0134729 A1 FIGURE 22

f f CH3 N-O Chs N-O S1 N h She S1 N H Seo. )=n st OC DMF )=n N is 2 HN O O.D2- Ns.Y HN O O - s! O1 O O1 OH s 52 S.

f CH N-O l w TFA, anisole .. Nh 1) Hexadecanedioic acid y=n 2 HATU, collidine

HN O O 2-k -a-samaalaamria

Patent Application Publication Jun. 14, 2007 Sheet 25 of 25

s NZ- s L - O -/8WdO)LO/HO,09+ .N&o+ON&oO. {?OOJLO-##OOJLO -

1s2 m - 2 NZ-- as 69 A Y-K f O **…!Oºr(C)?**)OY`C) US 2007/0134729 A1 Jun. 14, 2007

NOVELANTIBACTERAL AGENTS 0008 Antibacterial agents have proved to be important weapons in the fight against pathogenic bacteria. However, BACKGROUND OF THE INVENTION an increasing problem with respect to the effectiveness of 0001) 1. Field of the Invention antibacterial agents relates to the emergence of strains of 0002 This invention relates to novel multibinding com bacteria that are highly resistant to such agents. It would pounds (agents) that are antibacterial agents. The multib therefore be highly desirable to find antibacterial agents that inding compounds of the invention comprise from 2-10 are active against a broad spectrum of bacteria, in particular ligands covalently connected by a linker or linkers, wherein resistant strains. It would also be advantageous to discover each of said ligands in their monovalent (i.e., unlinked) state antibacterial agents that demonstrate high activity and selec have the ability to bind to an enzyme involved in cell wall tivity toward their targets, and are of low toxicity. biosynthesis and metabolism, a precursor used in the Syn 0009. The multibinding compounds of the present inven thesis of the bacterial cell wall and/or the cell surface and tion fulfill this need. thereby interfere with the synthesis and or metabolism of the cell wall. Preferably, the ligands are selected from the beta SUMMARY OF THE INVENTION lactam and/or glycopeptide class of antibacterial agents. 0010 This invention relates to novel multibinding com 0003. The invention also relates to pharmaceutical com pounds (agents) that are antibacterial agents. The multib positions comprising a pharmaceutically acceptable excipi inding compounds of the invention comprise from 2-10 ent and a therapeutically effective amount of one or more ligands covalently connected by a linker or linkers, wherein compound(s) of the invention, methods of using Such com each of said ligands in their monovalent (i.e., unlinked) state pounds and methods of preparing Such compounds. have the ability to bind to an enzyme involved in cell wall 0004 2. Background biosynthesis and metabolism, a precursor used in the Syn thesis of the bacterial cell wall and/or the bacterial cell 0005 Bacteria possess a rigid outer layer, the cell wall. surface and thereby interfere with the synthesis and/or The cell wall maintains the shape of the microorganism metabolism of the cell wall. Preferably, the ligands are which has a high internal osmotic pressure. Injury to the cell selected from the beta lactam and glycopeptide classes of wall (e.g. by ) or inhibition of the cell walls antibacterial agents. formation leads to lysis of the cell. 0011. The invention also relates to pharmaceutical com 0006 The cell wall contains a chemically distinct com positions comprising a pharmaceutically acceptable excipi plex polymer “mucopeptide' (“murein”, “') ent and a therapeutically effective amount of one or more consisting of polysaccharides and a highly cross-linked compound(s) of the invention, methods of using Such com polypeptide. The polysaccharides comprise an alternating pounds and methods of preparing Such compounds. copolymer of the amino Sugars N-acetylglucosamine and N-acetylmuramic acid, the latter being found only in bac 0012. Accordingly, in one aspect, this invention provides teria. To the N-acetylmuramic residues are attached pen a multibinding compound of Formula (I): tapeptides. The polysaccharide backbone of the cell wall is (L)(X). (I) formed by oligomerization of disaccharide pentapeptide precursors (lipid intermediate II) and is catalyzed an enzyme wherein: known as transglycosylase. The final rigidity of the cell wall 0013 p is an integer of from 2 to 10; is imparted by cross-linking of the peptide chains as a result of transpeptidation reactions by several bacterial enzymes 0014 q is an integer of from 1 to 20; one of which is known as peptidoglycan transpeptidase. 0015 each ligand, L, comprises a ligand domain capable 0007 One method by which antibacterial agents exert of binding to penicillin binding proteins, a transpeptidase their antibacterial activity is by inhibiting the transglycosy enzyme, a Substrate of a transpeptidase enzyme, a beta lase enzyme, thus interfering with the penultimate step in the lactamase enzyme, pencillinase enzyme, cephalosporinase synthesis of the bacterial cell wall. Although not wishing to enzyme, a transglycoslase enzyme, or a transglycosylase be bound by theory, it is believed that a glycopeptide, for enzyme Substrate; and example Vancomycin, binds with high affinity and specific 0016 X is a linker that may be the same or different at ity to N-terminal sequences (L-lysyl-D-alanyl-D-alanine in each occurrence; and pharmaceutically acceptable salts Vancomycin sensitive organisms) of peptidoglycan precur thereof provided that: sors known as lipid intermediate II. By binding to and sequestering these precursors, Vancomycin prevents their 0017 (i) all the ligands in a multibinding compound of utilization by the cell wall biosynthesis machinery. In a Formula (I) cannot be either an optionally substituted formal sense, therefore, Vancomycin inhibits the bacterial glycopeptide antibiotic, or an aglycone derivative of an transglycosylase that is responsible for adding lipid inter optionally Substituted glycopeptide antibiotic; mediate II subunits to growing peptidoglycan chains. This 0018 (ii) when p is 2 and q is 1 then at least one of the step preceeds the cross-linking transpeptidation step which ligands is a beta lactam antibiotic; and is inhibited by beta lactam antibiotics. It is believed that the B-lactam antibiotics bind to certain cell receptors (the peni 0019 (iii) when p is 2, q is 1, and one of the ligands is cillin binding proteins, “PBPs) which catalyze the Vancomycin attached to a linker via the C terminus, then transpeptidation reaction and other cell wall metabolic pro the other ligand cannot be attached to the linker cesses. The incomplete cell wall likely serves as a substrate via acylation of its alpha amino group. for autolytic enzymes in the cell wall and results in lysis if 0020 Preferably, q is less thanp in the multibinding the environment is isotonic. compounds of this invention. US 2007/0134729 A1 Jun. 14, 2007

0021. In a second aspect, this invention provides a multi wherein: Pyigy compound of Formula (I): (L)(X), (I) 0034 R is substituted alkyl, aryl aralkyl, or heteroaryl wherein: wherein each of said Substituent optionally links (a) to a linker via a covalent bond or R is a covalent bond that links 0022 p is an integer of from 2 to 10; (a) to a linker; and 0023 q is an integer of from 1 to 20; 0035) R' and Rare, independently of each other, alkyl or 0024 each ligand, L. is a beta lactam antibiotic, an at least one of R' and R is a covalent bond linking (a) to a optionally Substituted glycopeptide antibiotic, or an agly linker; cone derivative of an optionally substituted glycopeptide 0036 (ii) a compound of formula (b): antibiotic;

0.025 X is a linker that may be same or different at each (b) occurrence provided that: R5 0026 (i) all the ligands in a multibinding compound of R3-CO-NH Q Np Formula (I) cannot be. either an optionally substituted glycopeptide antibiotic, or an aglycone derivative of an N 21 R4 optionally Substituted glycopeptide antibiotic; O 0027 (ii) when p is 2 and q is 1 then at least one of the COOH ligands is a beta lactam antibiotic; and 0028 (iii) when p is 2, q is 1, and one of the ligands is wherein: Vancomycin attached to a linker via the C terminus, then the other ligand cannot be cefalexin attached to the linker 0037 one of P and Q is O, S, or -CH and the other via acylation of its alpha amino group. is —CH2—, 0029 Preferably, q is less than p: 0038) R is substituted alkyl, heteroarylalkyl, aralkyl, heterocyclylalkyl, or - C(R)=NOR" (where R is aryl, 0030 each ligand that is a beta lactam antibiotic is heteroaryl, or substituted alkyl; and R is alkyl or substituted selected from the group consisting of , penams, alkyl) wherein each of said substituent optionally links (b) to , , , , and a linker or R is a covalent bond that links (b) to a linker; and monobactam ring systems; and 0039) R' is hydrogen, alkyl, alkenyl, substituted alk 0031 each ligand that is a glycopeptide antibiotic is enylene, Substituted alkyl, halo, heteroarylalkyl, heterocy selected from the group consisting of Actaplanin, Actinodi clylalkyl, - SR" (where R is aryl, heteroaryl, heterocyclyl, din, Ardacin, Avoparcin, AZureomycin, A477, A35512, or cycloalkyl) or —CHSR' (where R is aryl, heteroaryl, A40926, A41030, A42867, A47934, A80407, A82846, A83850, A84575, A84428, AB-65, Balhimycin, Chloroer heterocyclyl, or cycloalkyl) wherein each of said substituent emomycin, Chloroorienticin, Chloropolysporin, Decapla optionally links (b) to a linker or R' is a covalent bond that nin, N-demethylvancomycin, Eremomycin, Galacardin, links (b) to a linker; Helvecardin. IZupeptin, Kibdelin, LL-AM374, Mannopep 0040 Riis hydrogen, hydroxy, or alkoxy: tin, MM45289, MM47756, MM47761, MM47921, MM47766, MM55260, MM55266, MM55270, MM56579, 0041 (iii) a compound of formula (c): MM56598, OA-7653, Oreenticin, Parvodicin. Ristocetin, Ristomycin, Synmonicin, , UK-68597,

UK-69542. UK-72051, optionally substituted Vancomycin, (c) and aglycone derivatives thereof. 0032 More preferably, each ligand that is a beta lactam antibiotic is selected from the group consisting of 0033 (i) a compound of formula (a):

(a) wherein: 0.042 T is S or CH: 0.043 R is alkyl; 0044) W is O, S, OCH , or CH; and R is -(alky O COOH lene)-NHC(R)=NH where R is a covalent bond linking (c) to a linker; or - W R is a covalent bond that links (c) to a linker; US 2007/0134729 A1 Jun. 14, 2007

0045 (iv) a compound of formula (d): 0053 Even more preferably, each ligand that is a beta lactam antibiotic is selected from the group consisting of 0054 (i) a compound of formula (a):

(d) (a) O

R l NH S R1 R2

N O COOH wherein: wherein: 0046 R and R are alkyl: 0055 R is: 0047) R' is selected from the group consisting of hydro gen, alkyl, Substituted alkyl, halo, aryl, heteroaryl, hetero R3NC o R.YC o cyclyl, aralkyl, heteroaralkyl, heterocyclylalkyl or \ / CH- \ / OCH —CHPSR' (where R is aryl, heteroaryl, heterocyclyl, or cycloalkyl) wherein each of said substituent optionally links OCH (d) to a linker orat, least one of RandR' is a covalent bond R3 o SC 19 that links (d) to a linker; or \ / Ri'O ti 0048 RandR' together with the carbon atoms to which NHR18 they are attached form an aryl, heteroaryl, cycloalkyl, Sub stituted cycloalkyl, or heterocyclyl ring of 4 to 7 ring atoms R3 / wherein one of the ring atoms optionally links (d) to a linker; * \ O 0049 (v) a compound of formula (e):

R20 = R2 = H OCH5

(e) R20 = R2 = Cl R20 = Cl and R2 = H

COORI9 COOR17 RSC7 o

\ / NHCO wherein: V 0050) R' is -SOH or -(alkylene)-COOH: 0051) R' is alkyl, substituted alkyl, haloalkyl, alkoxy, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, Substi tuted cycloalkyl, or heterocyclyl wherein each of said sub stituent optionally binds (e) to a linker or R'' is a covalent K)--NHCO bond that links (e) to a linker; and V 0052) R' is alkyl, acyl, or COC(R)=N OR'S wherein R'' is aryl, heteroaryl which optionally links (e) to a linker, and R' is -(alkylene)-COOR wherein R' is C O hydrogen or optionally links (e) to a linker or R' is a covalent bond that links (e) to a linker, and pharmaceutically acceptable salts thereof; US 2007/0134729 A1 Jun. 14, 2007

where: 0059 (ii) a compound of formula (b)): (b) 0056) R'' is a covalent bond that links the (a) group to a linker; R3-CO-NH 0057 one of R'' and R is hydrogen and the other is a covalent bond that links the (a) group to a linker, and where: 0058) R' and Rare methyl: 0060 R and R are:

R3 RNIf& ) -- - CHOCOCH, S

Ns N-CH,- ic-C-en R18 )-i- —CH

—CH

NHRI9

R18SC SCH SCH NS O N o s / - ( )-- Ns N -CH3 N - N-CHSO N N

18 CHOCONHR'. CHOCOCH, R’s Qg \ CH S

RISSg

()--o NHR19 RIS &| ) CHOCONHR' O NOCH

R18SC SCH NS / N N-CHCOOH CH-NHR19 US 2007/0134729 A1 Jun. 14, 2007

-continued R3 R4 —CHOCOCH, —CHOCH, H N - \ NOCH R17NH S

H3C N OH H3C N OH N N1 n N1 n \ O R18NH - S \ X NO-R19 -CHS lsN O -S lsN O X = halo

Y R19

- N 2 Nn-R 19 HC Y R R S "NQ \ R NR), RINH18 -SN N No1 R19 R17 SCH V O CH- N NHCO N 1 N-R N

CCO C2H5 Y 21 NZ

O 1N Y R3SN3 R31 Y- NR), 2 1N R (Note: the R group in the left column is paired with the R' in the right column) wherein:

0061 n is 0 or 1; m is 1-5; Z is CH or N: Y is H or halo: together with the nitrogen atom to which they are attached R is alkyl: R'' is a covalent bond that links the (b) group to form a heterocycloamino group; and R, R- and R are a linker; one of R'' and R' is hydrogen or alkyl; R and independently alkyl wherein one of R. R. R. R. is a R" are, independently of each other, hydrogen or alcyl; or covalent bond that links the (b) group to a linker; US 2007/0134729 A1 Jun. 14, 2007

0062 (iii) a compound of formula (c): where: 0064) R' is a covalent bond that links (d) to a linker; 0065) one of R and R is hydrogen, alkyl substituted (c) alkyl, or aralkyl, and other is a covalent bond that hinks (d) to a linker; R is alkyl; or 0066 (v) a compound of formula (e):

(e)

CH wherein R is a covalent bond lining (c) to a linker;

0063 (iv) a compound of formula (d): CH3 NEC

(d) Y SRa NHR22 wherein one of R and R’ is hydrogen and the other links (d) to a linker, and 0067 X is selected from a compound of formula:. —X"—Z-(Y" Z), X"— where R is: wherein 0068 m is an integer of from 0 to 20; OH O 0069 X at each separate occurrence is selected from the R25

N bond where R is as defined below: N N R26 24 0070 Z at each separate occurrence is selected from the R Yes R24 group consisting of alkylene, Substituted alkylene, cycloalkylene, Substituted cylcoalkylene, alkenylene, Sub stituted alkenylene, alkynylene, Substituted alynylene, cycloalkenylene, Substituted cycloalkenylene, arylene, het N eroarylene, heterocyclene, or a covalent bond; R23 0071 each Y at each separate occurrence is selected O from the group consisting of —O—, —C(O)—, O OC(O) , C(O)O , NR S(O)n , C(O)NR' NR C(O)—, NRC(O)NR' , -> ls NR'C(S)NR' , C(=NR). NR , NR N- R25 C(=NR) , OC(O) NR' , NR' C(O)-O-, R24 N=C(X) NR , NR' CCX)=N R25 P(O)(OR)-O-, -O-P(O)(OR") , S(O)CR'R" , S(O), NR' —NR' S(O), . —S S , and a cova R25 N lent bond; where n is 0, 1 or 2; and R, R and R" at each separate occurrence are selected from the group consisting O)-ris,N ( of hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, alkenyl, Substituted alkenyl, cycloalkenyl, Sub R24 stituted cycloalkenyl, alkynyl, Substituted alkynyl, aryl, het R24 eroaryl and heterocyclic. O 0072. In a second aspect, the invention relates to a method of treatment of mammals having a disease state that N(CH3)2 is treatable by antibacterial agents, comprising administer N ing a therapeutically effective amount of a compound of Formula (I), or a mixture of compounds of Formula (I), R23 thereto. 0073. In a third aspect, the invention relates to a phar maceutical composition comprising a therapeutically effec US 2007/0134729 A1 Jun. 14, 2007

tive amount of one or more compounds of Formula (I) or a of the library of ligands identified in (a) with the linker or pharmaceutically acceptable salt thereof, admixed with at mixture of linkers identified in (b) under conditions wherein least one pharmaceutically acceptable excipient. the complementary finctional groups react to form a cova 0074. In a fourth aspect, this invention is directed to lent linkage between said linker and at least two of said general synthetic methods for generating large libraries of ligands; and diverse multimeric compounds which multimeric com 0085 (d) assaying the multimeric ligand compounds pro pounds are candidates for possessing multibinding proper duced in (c) above to identify multimeric ligand compounds ties. The diverse multimeric compound libraries provided by possessing multibinding properties. this invention are synthesized by combining a linker or linkers with a ligand or ligands to provide for a library of 0086) The preparation of the multimeric ligand com multimeric compounds wherein the linker and ligand each pound library is achieved by either the sequential or con have complementary functional groups permitting covalent current combination of the two or more stoichiometric linkage. The library of linkers is preferably selected to have equivalents of the ligands identified in (a) with the linkers diverse properties such as Valency, linker length, linker identified in (b). Sequential addition is preferred when a geometry and rigidity, hydrophilicity or hydrophobicity, mixture of different ligands is employed to ensure het amphiphilicity, acidity, basicity and polarization. The library erodimeric or multimeric compounds are prepared. Concur of ligands is preferably selected to have diverse attachment rent addition of the ligands occurs when at least a portion of points on the same ligand, different finctional groups at the the multimer compounds prepared are homomultimeric same site of otherwise the same ligand, and the like. compounds. 0075. In a fifth aspect, this invention is directed to 0087. The assay protocols recited in (d) can be conducted libraries of diverse multimeric compounds which multim on the multimeric ligand compound library produced in (c) eric compounds are candidates for possessing multibinding above, or preferably, each member of the library is isolated properties. These libraries are prepared via the methods by preparative liquid chromatography mass spectrometry described above and permit the rapid and efficient evaluation (LCMS). of what molecular constraints impart multibinding proper 0088. In one of its composition aspects, this invention is ties to a ligand or a class of ligands targeting a receptor. directed to a library of multimeric ligand compounds which 0.076 Accordingly, in one of its method aspects, this may possess multivalent properties which library is prepared invention is directed to a method for identifying multimeric by the method comprising: ligand compounds possessing multibinding properties which 0089 (a) identifying a ligand or a mixture of ligands method comprises: wherein each ligand contains at least one reactive function 0.077 (a) identifying a ligand or a mixture of ligands ality; wherein each ligand contains at least one reactive function ality; 0090 (b) identifying a library of linkers wherein each linker in said library comprises at least two finctional groups 0078 (b) identifying a library of linkers wherein each having complementary reactivity to at least one of the linker in said library comprises at least two functional reactive functional groups of the ligand; and groups having complementary reactivity to at least one of the reactive functional groups of the ligand; 0091 (c) preparing a multimeric ligand compound library by combining at least two stoichiometric equivalents 0079 (c) preparing a multimeric ligand compound of the ligand or mixture of ligands identified in (a) with the library by combining at least two stoichiometric equivalents library of linkers identified in (b) under conditions wherein of the ligand or mixture of ligands identified in (a) with the the complementary functional groups react to form a cova library of linkers identified in (b) under conditions wherein lent linkage between said linker and at least two of said the complementary finctional groups react to formin a cova ligands. lent linkage between said linker and at least two of said ligands; and 0092. In another of its composition aspects, this invention is directed to a library of multimeric ligand compounds 0080 (d) assaying the multimeric ligand compounds pro which may possess multivalent properties which library is duced in (c) above to identify multimneric ligand.com prepared by the method comprising: pounds possessing multibinding properties. 0093 (a) identifying a library of ligands wherein each 0081. In another of its method aspects, this invention is directed to a method for identifying multimeric ligand ligand contains at least one reactive functionality; compounds possessing multibinding properties which 0094 (b) identifying a linker or mixture of linkers method comprises: wherein each linker comprises at least two functional groups 0082 (a) identifying a library of ligands wherein each having complementary reactivity to at least one of the ligand contains at least one reactive functionality; reactive finctional groups of the ligand; and 0.083 (b) identifying a linker or mixture of linkers 0095 (c) preparing a multimeric ligand compound wherein each linker comprises at least two functional groups library by combining at least two stoichiometric equivalents having complementary reactivity to at least one of the of the library of ligands identified in (a) with the linker or mixture of linkers identified in (b) under conditions wherein reactive flinctional groups of the ligand; the complementary functional groups react to form a cova 0084 (c) preparing a multimeric ligand compound lent linkage between said linker and at least two of said library by combining at least two stoichiometric equivalents ligands. US 2007/0134729 A1 Jun. 14, 2007

0096. In a preferred embodiment, the library of linkers molecular constraints imparting multibinding properties to employed in either the methods or the library aspects of this the multimeric compound or compounds found in said first invention is selected from the group comprising flexible iteration; linkers, rigid linkers, hydrophobic linkers, hydrophilic link ers, linkers of different geometry, acidic linkers, basic link 0105 (f) evaluating what molecular constraints imparted ers, linkers of different polarization and or polarizability and enhanced multibinding properties to the multimeric com amphiphilic linkers. For example, in one embodiment, each pound or compounds found in the second collection or of the linkers in the linker library may comprise linkers of iteration recited in (e) above: different chain length and/or having different complemen 0106 (g) optionally repeating steps (e) and (f) to further tary reactive groups. Such linker lengths can preferably elaborate upon said molecular constraints. range from about 2 to 100 A. 0.107 Preferably, steps (e) and (f) are repeated at least 0097. In another preferred embodiment, the ligand or two times, more preferably at from 2-50 times, even more mixture of ligands is selected to have reactive functionality preferably from 3 to 50 times, and still more preferably at at different sites on said ligands in order to provide for a least 5-50 times. range of orientations of said ligand on said multimeric ligand compounds. Such reactive functionality includes, by BRIEF DESCRIPTION OF THE DRAWINGS way of example, carboxylic acids, carboxylic acid halides, carboxyl esters, amines, halides, pseudohalides, isocyan 0.108 FIG. 1 illustrates examples of multibinding com ates, vinyl unsaturation, ketones, aldehydes, thiols, alcohols, pounds comprising 2 ligands attached in different formats to anhydrides, boronates, and precursors thereof. It is under a linker. stood, of course, that the reactive functionality on the ligand 0.109 FIG. 2 illustrates examples of multibinding com is selected to be complementary to at least one of the pounds comprising 3 ligands attached in different formats to reactive groups on the linker so that a covalent linkage can a linker. be formed between the linker and the ligand. 0110 FIG. 3 illustrates examples of multibinding com 0098. In other embodiments, the multimeric ligand com pounds comprising 4 ligands attached in different formats to pound is homomeric (i.e., each of the ligands is the same, a linker. although it may be attached at different points) or hetero meric (i.e., at least one of the ligands is different from the 0.111 FIG. 4 illustrates examples of multibinding com other ligands). pounds comprising >4 ligands attached in different formats to a linker. 0099. In addition to the combinatorial methods described herein, this invention provides for an iterative process for 0112 FIGS. 5, 6A, and 6B disclose some representative rationally evaluating what molecular constraints impart compounds of formula (a) and (b). multibinding properties to a class of multimeric compounds 0113 FIGS. 7-10 disclose examples of multibinding or ligands targeting a receptor. Specifically, this method compounds comprising 2ligands attached in different for aspect is directed to a method for identifying multimeric matS. ligand compounds possessing multibinding properties which method comprises: 0114 FIGS. 11-23 illustrate synthesis of compounds of Formula (I). 0100 (a) preparing a first collection or iteration of mul timeric compounds which is prepared by contacting at least DETAILED DESCRIPTION OF THE two stoichiometric equivalents of the ligand or mixture of INVENTION ligands which target a receptor with a linker or mixture of linkers wherein said ligand or mixture of ligands comprises Definitions at least one reactive functionality and said linker or mixture of linkers comprises at least two functional groups having 0115 This invention is directed to multibinding com complementary reactivity to at least one of the reactive pounds that are antibacterial agents and pharmaceutical functional groups of the ligand wherein said contacting is compositions containing Such compounds. When discussing conducted under conditions wherein the complementary Such compounds, and compositions the following terms functional groups react to form a covalent linkage between have the following meanings unless otherwise indicated. said linker and at least two of said ligands; Any undefined terms have their art recognized meanings. 0101 (b) assaying said first collection or iteration of 0.116) The term “alkyl refers to a monoradical branched multimeric compounds to assess which if any of said mul or unbranched saturated hydrocarbon chain preferably hav timeric compounds possess multibinding properties; ing from 1 to 40 carbon atoms, more preferably 1 to 1 carbon atoms, and even more preferably 1 to 6 carbon atoms. This 0102 (c) repeating the process of (a) and (b) above until term is exemplified by groups such as methyl, ethyl, n-pro at least one multimeric compound is found to possess pyl, iso-propyl. n-butyl, iso-butyl, n-hexyl, n-decyl, tetrade multibinding properties; cyl, and the like. 0103 (d) evaluating what molecular constraints imparted 0117 The term “substituted alkyl” refers to an alkyl multibinding properties to the multimeric compound or group as defined above, having from 1 to 5 substituents, and compounds found in the first iteration recited in (a)-(c) preferably 1 to 3 substituents, selected from the group above; consisting of alkoxy, Substituted alkoxy, cycloalkyl, Substi 0104 (e) creating a second collection or iteration of tuted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, multimeric compounds which elaborates upon the particular acyl, acylamino, acyloxy, amino, Substituted amino, ami US 2007/0134729 A1 Jun. 14, 2007 noacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halo 0.124. The term “substituted alkoxy' refers to the groups gen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thio substituted alkyl-O substituted alkenyl-O substituted aryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, cycloalkyl-O-, Substituted cycloalkenyl-O-, and Substi thioalkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, tuted alkynyl-O where substituted alkyl, substituted alk heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, enyl, Substituted cycloalkyl, Substituted cycloalkenyl and alkoxyamino, nitro, guanidine. —C(=NRa)NHR' (where substituted alkynyl are as defined herein. R and Rare independently selected from hydrogen, alkyl, 0.125 The term “alkenyl refers to a monoradical of a aryl, aralkyl, heteroaryl, or heteroaralkyl, NHSONHR branched or unbranched unsaturated hydrocarbon group (where R is hydrogen, alkyl, aryl, aralkyl, heteroaryl, or preferably having from 2 to 40 carbon atoms, more prefer heteroaralkyl) —SO-alkyl, -SO-substituted alkyl, - SO aryl, SO-heteroaryl, -SO-alkyl, -SO-substituted ably 2 to 10 carbon atoms and even more preferably 2 to 6 alkyl, -SO2-aryl and 13 SO-heteroaryl. This term is carbon atoms and having at least 1 and preferably from 1-6 exemplified by groups such as hydroxymethyl, hydroxy sites of vinyl unsaturation. Preferred alkenyl groups include ethyl, hydroxypropyl, 2-aminoethyl, 3-aminopropyl. ethenyl ( CH=CH-), n-propenyl.( CH-CH=CH-), iso 2-methylaminoethyl, 3-dimethylaminopropyl, 2-sulfona propenyl ( C(CH)=CH), and the like. midoethyl, 2-carboxyethyl, and the like. 0.126 The term “substituted alkenyl refers to an alkenyl group as defined above having from 1 to 5 substituents, and 0118. The term “al kylene' refers to a diradical of a preferably 1 to 3 substituents, selected from the group branched or unbranched saturated hydrocarbon chain, pref consisting of alkoxy, Substituted alkoxy, cycloalkyl, Substi erably having from 1 to 40 carbon atoms, more preferably 1 tuted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, to 10 carbon atoms and even more preferably 1 to 6 carbon acyl, acylamino, acyloxy, amino, Substituted amino, ami atoms. This term is exemplified by groups such as methylene noacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halo (—CH2—), ethylene (-CH2CH2—), the propylene isomers gen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thio (e.g., —CH2CH2CH2— and —CH(CH)CH2—) and the aryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, like. thioalkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, 0119) The term “substituted alkylene' refers to: heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, 0120 (a) an alkylene group, as defined above, having alkoxyamino, nitro. —SO-alkyl, -SO-substituted alkyl, from 1 to 5 substituents, and preferably 1 to 3 substituents, —SO-aryl, -SO-heteroaryl, —SO-alkyl, -SO-substi Selected from the group consisting of alkoxy, Substituted tuted alkyl, -SO-aryl and —SO-heteroaryl. alkoxy, cycloalkyl, Substituted cycloalkyl, cycloalkenyl, 0127. The term “alkenylene' refers to a diradical of a Substituted cycloalkenyl, acyl, acylamino, acyloxy, branched or unbranched unsaturated hydrocarbon group amino, Substituted amino, aminoacyl, aminoacyloxy, preferably having from 2 to 40 carbon atoms, more prefer oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, ably 2 to 10 carbon atoms and even more preferably 2 to 6 thioketo, carboxyl, carboxylalkyl, thioaryloxy, thiohet carbon atoms and having at least 1 and preferably from 1-6 eroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, Substi sites of vinyl unsaturation. This term is exemplified by tuted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, groups such as ethenylene (-CH=CH-), the propenylene heterocyclic, heterocyclooxy, hydroxyamino, isomers (e.g., —CH-CH=CH , —C(CH)=CH , and alkoxyamino, nitro. —SO-alkyl, -SO-substituted alkyl, the like. —SO-aryl, -SO-heteroaryl, -SO-alkyl, -SO-substi 0128. The term “substituted alkenylene' refers to an tuted alkyl, -SO-aryland—SO-heteroaryl. Addition alkenylene group as defined above having from 1 to 5 ally, Such Substituted alkylene groups include those where substituents, and preferably from 1 to 3 substituents, 2 Substituents on the alkylene group are fused to form one selected from the group consisting of alkoxy, Substituted or more cycloalkyl, Substituted cycloalkyl, cycloalkenyl, alkoxy, cycloalkyl, Substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heterocyclic or heteroaryl Substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, groups fused to the. alkylene group. Preferably such fused Substituted amino, aminoacyl aminoacyloxy, oxyaminoacyl, groups contain from 1 to 3 fused ring structures; azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, 0121 (b) an alkylene group as defined above wherein one carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocy or more carbons atoms, is replaced by oxygen, Sulfur, and clooxy, thiol, thioalkoxy, Substituted thioalkoxy, aryl, ary —NR— where R is hydrogen, substituted alkyl, loxy, heteroaryl, heteroaryloxy, heterocyclic, heterocy cycloalkyl, alkenyl cycloalkenyl, alkynyl, aryl, heteroaryl clooxy, hydroxyamino, alkoxyamino, nitro. —SO-alkyl, and heterocyclic. —SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, 0122) The term "alkaryl' or “aralkyl refers to the groups —SO-alkyl, -SO-substituted alkyl, -SO2-aryl and -alkylene-aryl and -substituted alkylene-aryl where alky —SO-heteroaryl. Additionally, such substituted alkenylene lene, substituted alkylene and aryl are defined herein. Such groups include those where 2 substituents on the alkenylene alkaryl groups are exemplified by benzyl, phenethyl and the group are fused to form one or more cycloalkyl, Substituted like. cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, aryl, het 0123 The term “alkoxy” refers to the groups alkyl-O , erocyclic or heteroaryl groups fused to the alkenylene group. alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alky 0129. The term “alkynyl refers to a monoradical of an nyl-O-, where alkyl, alkenyl, cycloalkyl, cycloalkenyl, and unsaturated hydrocarbon preferably having from 2 to 40 alkynyl are as defined herein. Preferred alkoxy groups are carbon atoms, more preferably 2 to 20 carbon atoms and alkyl-O- and include, by way of example, methoxy, even more preferably 2 to 6 carbonatoms and having at least ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec 1 and preferably from 1-6 sites of acetylene (triple bond) butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the unsaturation. Preferred alkynyl groups include ethynyl like. (—C CH), propargyl (—CH2C=CH) and the like. US 2007/0134729 A1 Jun. 14, 2007

0130. The term “substituted alkynyl refers to an alkynyl alkyl, Substituted alkyl, alkenyl, Substituted alkenyl, aryl, group as defined above having from 1 to 5 substituents, and heteroaryl and heterocyclic are as defined herein. preferably 1 to 3 substituents, selected from the group consisting of alkoxy, Substituted alkoxy, cycloalkyl, Substi 0.137 The term “aminoacyloxy' or “alkoxycarbony tuted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, lamino” refers to the group —NRC(O)OR where each R is acyl, acylamino, acyloxy, amino, Substituted amino, ami independently hydrogen, alkyl, Substituted alkyl, aryl, het noacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halo eroaryl, or heterocyclic wherein alkyl, substituted alkyl, gen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thio aryl, heteroaryl and heterocyclic are as defined herein. aryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, 0.138. The term “acyloxy' refers to the groups alkyl thioalkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, C(O)O substituted alkyl-C(O)O cycloalkyl-C(O)O— heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, substituted cycloalkyl-C(O)O , aryl-C(O)O—, heteroaryl alkoxyamino, nitro. —SO-alkyl, -SO-substituted alkyl, C(O)O—, and heterocyclic-C(O)O— wherein alkyl, substi —SO-aryl, -SO-heteroaryl, -SO-alkyl, -SO-substi tuted alkyl, cycloalkyl, Substituted cycloalkyl, aryl, het tuted alkyl, -SO-aryl, and —SO-heteroaryl. eroaryl, and heterocyclic are as defined herein. 0131 The term “alkynylene' refers to a diradical of an 0.139. The term “aryl” refers to an unsaturated aromatic unsaturated hydrocarbon preferably having from 2 to 40 carbocyclic group of from 6 to 20 carbon atoms having a carbon atoms, more preferably 2 to 10 carbon atoms and single ring (e.g., phenyl) or multiple condensed (fused) rings even more preferably 2 to 6 carbon atoms and having at least (e.g., naphthyl or anthryl). The aryl group may optionally be 1 and preferably from 1-6 sites of acetylene (triple bond) fuised to a heterocyclic or cycloalkyl group. Preferred unsaturation. Preferred alkynylene groups include ethy aryls.include phenyl, naphthyl and the like. Unless other nylene ( -C=C- ), propargylene (—CH2C=C ) and the wise constrained by the definition for the aryl substituent, like. Such aryl groups can optionally be substituted with from 1 to 5 suhstituents, preferably 1 to 3 substituents, selected 0132) The term “substituted alkynylene' refers to an from the group consisting of acyloxy, hydroxy, thiol, acyl, alkynylene group as defined above having from 1 to 5 alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituents, and preferably 1 to 3 substituents, selected substituted alkyl, substituted alkoxy, substituted alkenyl, from the group consisting of alkoxy, Substituted alkoxy, substituted alkynyl, substituted cycloalkyl, substituted cycloalkyl, Substituted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, amino, Substituted amino, aminoacyl, acy cycloalkenyl, acyl, acylamino, acyloxy, amino, Substituted lamino, Sulfonylamino, alkaryl, aryl, aryloxy, azido, car amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, boxyl, carb6Xylalkyl, cyano, halo, nitro, heteroaryl, het cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxy eroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, lalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, oxyacylanino, thioalkoxy, Substituted thioalkoxy, thioary thiol, thioalkoxy, Substituted thioalkoxy, aryl, aryloxy, het loxy, thioheteroaryloxy, —SO-alkyl, -SO-substituted eroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, alkyl, -SO-aryl, -SO-heteroaryl,- —SO-alkyl, - SO hydroxyamino, alkoxyamino, nitro. —SO-alkyl, -SO-Sub substituted alkyl, -SO-aryl, -SO-heteroaryl and triha stituted alkyl, —SO-aryl, -SO-heteroaryl, -SO-alkyl, lomethyl. Preferred aryl substituents include alkyl, alkoxy, —SO-substituted alkyl, -SO-aryland —SO-heteroaryl halo, cyano, nitro, trihalomethyl, and thioalkoxy. 0133) The term “acyl refers to the groups HC(O) , alkyl-C(O)—, substituted alkyl-C(O)—, alkenyl-C(O)— 0140. The term “aryloxy' refers to the group aryl-O substituted alkenyl-C(O)—, cycloalkyl-C(O)—, substituted wherein the aryl group is as defined above including option cycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted ally Substituted aryl groups as also defined above. cycloalkenyl-C(O)—, aryl-C(O)—, heteroaryl-C(O)— and 0141 The term "amino” refers to the group —NH. heterocyclic-C(O)— where alkyl, substituted alkyl...alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, 0142. The term “substituted amino” refers to the group —NRR where each R is independently selected from the cycloalkenyl, Substituted cycloalkenyl, aryl, heteroaryl and group consisting of hydrogen, alkyl, Substituted alkyl, acyl, heterocyclic are as defined herein. cycloalkyl, Substituted cycloalkyl, alkenyl, Substituted alk 0134) The term “acylamino” or “aminocarbonyl refers enyl, cycloalkenyl, Substituted cycloalkenyl, alkynyl. Sub to the group —C(O)NRR where each R is independently stituted alkynyl, aryl, heteroaryl and heterocyclic provided hydrogen, alkyl, Substituted alkyl, aryl, heteroaryl, hetero that both Rs are not hydrogen. cyclic or where both R groups are joined to form a hetero cyclic group (e.g., morpholino) wherein alkyl, Substituted 0143) The term “carboxyalkyl or “alkoxycarbonyl alkyl, aryl, heteroaryland heterocyclic are as defined herein. refers to the groups ' C(O)O-alkyl”, “ C(O)O-substi tuted alkyl”, “ C(O)O-cycloalkyl”, “ C(O)O-substituted 0135 The term "sulfonylamino” refers to the group cycloalkyl”, “ C(O)C-alkenyl', '-C(O)O-substituted alk —NRSOR" where R is hydrogen, alkyl, substituted alkyl, enyl”, “ C(O)O-alkynyl' and “ C(O)O-substituted alky aralkyl, or heteroaralkyl, and R is alkyl, substituted alkyl, nyl' where alkyl, substituted alkyl, cycloalkyl, substituted amino, or Substituted amino wherein alkyl, Substituted alkyl, cycloalkyl, alkenyl, Substituted alkenyl, alkynyl and Substi aralkyl, heteroaralkyl and Substituted amino are as defined tuted alkynyl are as defined herein. herein. 0144. The term “cycloalkyl refers to cyclic alkyl groups 0136. The term “aminoacyl refers to the group —NR of from 3 to 20 carbon atoms having a single cyclic ring or C(O)R where each R is independently hydrogen, alkyl, multiple condensed rings, said cycloalkyl group may option Substituted alkyl, alkenyl, Substituted alkenyl, amino, Sub ally be fused to an aryl or heteroaryl group. Such cycloalkyl stituted amino, aryl, heteroaryl, or heterocyclic wherein groups include, by way of example, single ring structures US 2007/0134729 A1 Jun. 14, 2007

Such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy. and the like, or multiple ring structures such as adamantanyl. Such heteroaryl groups can have a single ring (e.g., pyridyl and the like. or furyl) or multiple condensed rings (e.g., indolizinyl or 0145 The term “substituted cycloalkyl” refers to benzothienyl). Preferred heteroaryls include pyridyl, pyrro cycloalkyl groups having from 1 to 5 substituents, and lyl and furyl. preferably 1 to 3 substituents, selected from the group 0150. The term “heteroaryloxy” refers to the group het consisting of alkyl, Substituted alkyl, alkenyl, Substituted eroaryl-O-. alkenyl, alkoxy, Substituted alkoxy, cycloalkyl, Substituted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, acyl, 0151. The term “heterocycle” or “heterocyclyl refers to acylamino, acyloxy, amino, Substituted amino, aminoacyl, a monoradical Saturated or unsaturated group having a single aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, ring or multiple condensed rings, from 1 to 40 carbon atoms hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioary and from 1 to 10 hetero atoms, preferably 1 to 4 heteroat loxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thio oms, selected from nitrogen, Sulfur, phosphorus, and/or alkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, oxygen within the ring and further wherein one, two, or three heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, of the ring carbon atoms may optionally be replaced with a alkoxyamino, nitro. —SO-alkyl, 13 SO-substituted alkyl, carbonyl group (i.e., a keto group). The heterocycle group —SO-aryl, -SO-heteroaryl, -SO-alkyl, -SO-substi may optionally fused to an aryl or heteroaryl ring. Unless tuted alkyl, -SO-aryl and —SO-heteroaryl. otherwise constrained by the definition for the heterocyclic Substituent, Such heterocyclic groups can be optionally 0146 The term “cycloalkenyl refers to cyclic alkenyl substituted with 1 to 5, and preferably 1 to 3 substituents, groups of from 4 to 20 carbon atoms having a single cyclic selected from the group consisting of alkyl, acyloxy, alkyl, ring and at least one point of internal unsaturation. Examples Substituted alkyl, alkoxy, Substituted alkoxy, cycloalkyl, of Suitable cycloalkenyl groups include, for instance, Substituted cycloalkyl, cycloalkenyl, Substituted cycloalk cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-enyl and the enyl, acyl, acylamino, acyloxy, amino, Substituted amino, like. aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, 0147 The term “substituted cycloalkenyl refers to halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, cycloalkenyl groups having from 1 to 5 substituents, and thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, preferably 1 to 3 substituents, selected from the group thioalkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, consisting of alkyl, substituted alkyl, alkenyl, substituted heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkenyl, alkoxy, Substituted alkoxy, cycloalkyl, Substituted alkoxyamino, nitro. —SO-alkyl, -SO-substituted alkyl, cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, acyl, —SO-aryl, -SO-heteroaryl, -SO-alkyl, -SO-substi acylamino, acyloxy, amino, Substituted amino, aminoacyl, tuted alkyl, -SO-aryl and —SO-heteroaryl. Such hetero aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, cyclic groups can have a single ring or multiple condensed hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioary rings. Preferred heterocyclics include morpholino, piperidi loxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thio nyl, and the like. alkoxy, Substituted thioalkoxy, aryl, aryloxy, heteroaryl, 0152 Examples of heteroaryls and heterocycles include, heteroaryloxy, heterocyclic, heterocyclooxy. but are not limited to, pyrrole, thiophene, furan imidazole, hydroxyamino, alkoxyamino, nitro. —SO-alkyl, -SO-Sub pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, stituted alkyl, -SO-aryl, -SO-heteroaryl, -SO-alkyl, indolizine, isoindole, indole, indazole, purine, quinolizine, —SO-substituted alkyl, -SO-aryl and —SO-heteroaryl. isoquinoline, quinoline, phthalazine, naphthylpyridine, qui 0148. The term “halo” or “halogen” refers to fluoro, noxaline, quinazoline, cinnoline, pteridine, carbazole, car chloro, bromo and iodo. boline, phenanthridine, acridine, phenanthroline, isothiaz ole, phenazine, isoxazole, phenoxazine, phenothiazine, 014.9 The term "heteroaryl refers to an aromatic group imidazolidine, imidazoline, pyrrolidine, piperidine, pipera of from 1 to 15 carbonatoms and 1 to 4 heteroatoms selected Zine, indoline, morpholine, tetrahydrofuiranyl, tetrahy from oxygen, nitrogen and Sulfur within at least one ring (if drothiophene, and the like as well as N-alkoxy-nitrogen there is more than one ring). The heteroaryl ring may containing heterocycles. optionally be fused to a cycloalkyl or heterocyclyl ring. Unless otherwise constrained by the definition for the het 0.153 “Heterocycloamino” means a saturated, unsatur eroaryl Substituent, Such heteroaryl groups can be optionally ated, bridged or unbridged, monovalent cyclic group of 4 to substituted with 1 to 5 substituents, preferably 1 to 3 8 ring atoms, wherein at least one ring atom is N and Substituents, selected from the group consisting of acyloxy, optionally contains one or two additional ring heteroatoms hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, selected from the group consisting of N, O, or S(O)n (where cycloalkyl, cycloalkenyl, substituted alkyl, substituted n is an integer from 0 to 2), the remaining ring atoms being alkoxy, substituted alkenyl, substituted alkynyl, substituted C, where one or two Catoms may optionally be replaced by cycloalkyl, Substituted cycloalkenyl, amino, Substituted a carbonyl group. The heterocycloamino ring may be fused amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, to a cycloalkyl, aryl or heteroaryl ring, and it may be carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl, het optionally substituted with one or more substituents, pref eroaryloxy, heterocyclic, heterocyclooxy, aminoacyloxy, erably one or two substituents, selected from alkyl, substi oxyacylamino, thioalkoxy, Substituted thioalkoxy, thioary tuted alkyl, cycloalkyl, aryl, araiyl, heteroaryl, het loxy, thioheteroaryloxy, —SO-alkyl, -SO-substituted eroaralkyl, halo, cyano, acyl, amino, Substituted amino, alkyl, -SO-aryl, -SO-heteroaryl, —SO-alkyl, - SO acylamino, —OR (where R is hydrogen, alkyl, alkenyl, substituted alkyl, -SO-aryl, -SO-heteroaryl and triha cycloalkyl, acyl, aryl, heteroaryl, aralkyl, or heteroaralkyl). lomethyl. Preferred heteroaryl substituents include alkyl, or —S(O)nR where n is an integer from 0 to 2 and R is US 2007/0134729 A1 Jun. 14, 2007 hydrogen (provided that n is 0), alkyl, alkenyl, cycloalkyl, tuted cycloalkyl amine, trisubstituted cycloalkyl amines, amino, heterocyclo, aryl, heteroaryl, aralkyl, or het cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalk eroaralkyl). More specifically the term heterocycloamino enyl) amines, Substituted cycloalkenyl amiines, disubsti includes, but is not limited to, pyrrolidino, piperidino, mor tuted cycloalkenyl amine, trisubstituted cycloalkenyl pholino, piperazino, indolino, quinuclidine, or thiomor amines, aryl amines, diaryl amines, triaryl amines, het pholino, and the derivatives thereof. eroaryl amiines, diheteroaryl amines, triheteroarylainines, heterocyclic amnines, diheterocyclic amiines, triheterocy 0154) The term "heterocyclooxy' refers to the group clic amines, mixed di- and tri-amines where at least two of heterocyclic-O . the substituents on the amine are different and-are selected 0155 The term “thioheterocyclooxy” refers to the group from the group consisting of alkyl, Substituted alkyl, alk heterocyclic-S-. enyl, Substituted alkenyl, cycloalkyl, Substituted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, aryl, heteroaryl, het 0156 The term "oxyacylamino” or “aminocarbonyloxy” erocyclic, and the like. Also included are amiines where the refers to the group —OC(O)NRR where each R is indepen two or three Substituents, together with the amino nitrogen, dently hydrogen, alkyl Substituted alkyl, aryl, heteroaryl, or form a heterocyclic or heteroaryl group. Examples of Suit heterocyclic wherein alkyl, substituted alkyl aryl, heteroaryl able amines include, by way of example only, isopropy and heterocyclic are as defined herein. lamine, trimethyl amine, diethyl amine, tri(iso-propyl) 0157 The term “spiro-attached cycloalkyl group' refers amine, tri(n-propyl) amine, ethanolamine, 2-dimethylami to a cycloalkyl group joined to another ring via one carbon noethanol, tromethamine, lysine, arginine, histidine, caf atom common to both rings. feine, procaine, hydrabamine, choline, betaine, ethylenedi amine, glucosamine, N-alkylglucamines, theobromine, 0158. The term “thiol” refers to the group -SH. purines, piperazine, piperidine, morpholine, N-ethylpiperi 0159. The term “thioalkoxy” or “alkylthio” refers to the dine, and the like. It should also be understood that other group —S-alkyl. carboxylic acid derivatives would be useful in the practice of this invention, for example, carboxylic acid amides, includ 0160 The term “substituted thioalkoxy” refers to the ing carboxamides, lower alkyl carboxamides, dialkyl car group —S-Substituted alkyl. boxamides, and the like. 0161 The term “thioaryloxy' refers to the group aryl-S 0166 Pharmaceutically acceptable acid addition salts wherein the aryl group is as defined above including option may be prepared from inorganic and organic acids. Salts ally Substituted aryl-groups also defined above. derived from inorganic acids include hydrochloric. acid, 0162 The term “thioheteroaryloxy' refers to the group hydrobromic acid, Sulfuric acid, nitric acid, phosphoric acid, heteroaryl-S wherein the heteroaryl group is as defined and the like. Salts derived from organic acids include acetic above including optionally Substituted aryl groups as also acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, defined above. mtalic acid, malonic acid, Succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cin 0163 As to any of the above groups which contain one or namic acid, mandelic acid, methanesulfonic acid, ethane more Substituents, it is understood, of course, that Such Sulfonic acid, p-toluene-Sulfonic acid, Salicylic acid, and the groups do not contain any Substitution or Substitution pat like. terns which are sterically impractical and/or synthetically non-feasible. In addition, the compounds-of this invention 0.167 The term “pharmaceutically-acceptable cation' include all stereochemical isomers arising from the Substi refers to the cation of a pharmaceutically-acceptable salt. tution of these compounds. 0.168. The term “protecting group' or “blocking group' 0164. The term “pharmaceutically-acceptable salt” refers refers to any group which when bound to one or more to salts which retain the biological effectiveness and prop hydroxyl, thiol, amino or carboxyl groups of the compounds erties of the multibinding compounds of this invention and (including intermediates thereof) prevents reactions from which are not biologically or otherwise undesirable. In many occurring at these groups and which protecting group can be cases, the multibinding compounds of this invention are removed by conventional chemical or enzymatic steps to capable of forming acid and/or base salts by virtue of the reestablish the hydroxyl, thiol, amino or carboxyl group presence of amino and/or carboxyl groups or groups similar (See., T. W. Greene and P. G. H. Wuts, “Protective Groups thereto. in Organic Synthesis", 2" Ed.). The particular removable blocking group employed is not critical and preferred 0165 Pharmaceutically-acceptable base addition salts can removable hydroxyl blocking groups include conventional be prepared from inorganic and organic bases. Salts derived Substituents such as allyl, benzyl, acetyl, chloroacetyl, from inorganic bases, include by way of example only, thiobenzyl, benzylidine, phenacyl, t-butyl-diphenylsilyl and Sodium, potassium, lithium, ammonium, calcium and mag any other group that can be introduced chemically onto a nesium salts. Salts derived from organic bases include, but hydroxyl functionality and later selectively removed either are not limited to, salts of primary, secondary and tertiary by chemical or enzymatic methods in mild conditions com amines, such as alkyl amines, dialkyl amines, trialkyl patible with the nature of the product. Preferred removable amines, Substituted alkyl amines, di(Substituted alkyl) thiol blocking groups include disulfide groups, acyl groups, amines, tri(Substituted alkyl)amines, alkenyl amines, dialk benzyl groups, and the like. enyl armines, trialkenyl amines, Substituted alkenyl amines, di(substituted alkenyl) amines, tri(substituted alkenyl) 0.169 Preferred removable amino blocking groups amines, cycloalkyl amines, di(cycloalkyl) amines, tri(cy include conventional Substituents such as t-butyoxycarbonyl cloalkyl) amines, Substituted cycloalkyl amines, disubsti (t-BOC), benzyloxycarbonyl (CBZ), fluorenylmethoxy-car US 2007/0134729 A1 Jun. 14, 2007 13 bonyl (FMOC), allyloxycarbonyl (ALOC), and the like which can be removed by conventional conditions compat ible with the nature of the product. 0170 Preferred carboxyl protecting groups include esters Such as methyl, ethyl, propyl, t-butyl etc. which can be removed by mild conditions compatible with the nature of the product. 0171 The term “optional” or “optionally” means that the 0.174 The B-lactam antibiotics are classified into the Subsequently described, event, circumstance or Substituent , , carbapenems, oxacephems, car may or may not occur, and that the description includes bacephems, and and include drugs such as instances where said event or circumstance occurs and Penicillin G, Penicillin V. Methicillin, , Cloxacil instances where it does not. lin, Dicloxacillin, , Ampicillin, Amoxicillin, Car benicillin, Carbenicillin indanyl. Ticarcillin, Mezlocillin, 0172 The term “ligand’ or “ligands' as used herein Cephalothin, , Cephalexin, denotes a compound that is a binding partner for penicillin , Cefamandole, Cefoxitin, Cefaclor, Cefuroxime, binding proteins, a pencillinase enzyme, a cephalosporinase Cefuroxime axetil, , Cefonicid, , Cefo enzyme, a transpeptidase enzyme, a Substrate of a transpep ranide, , proxetil, Ceftizoxime, tidase enzyme, a beta-lactamase enzyme, a transglycoslase , , , Imi enzyme, or a transglycosylase enzyme Substrate and is , , , , L-695256, bound thereto by complementarity. The specific region or GV-143253, Sanifitrinem, Fropenem, Lactivicin, BO-2727, regions of the ligand that is (are) recognized by the penicillin MEN-10700, Ro-48-8724, Cefosilis, SB-216477, S-4661, binding proteins, a pencillinase enzyme, a cephalosporinase GG-326, BLA-857, PGE-8335534, PGE-542860, enzyme, a transpeptidase enzyme, a Substrate of a transpep LB-10522, GV 129606, BO-2052A, CS-834, MK-826, tidase enzyme, a beta-lactamase enzyme, a transglycoslase YH-1226, YM-40220, MDL-63908, FCE251.99, enzyme, or a transglycosylase enzyme substrate is desig , TOC-50, TOC-39, TOC-29, E-1 101, nated as the “ligand domain'. A ligand may be either capable Sulopenem, DU-6681, MC-02479, , Carunonam, of binding to its target by itself, or may require the presence Ro-25-0534, SUN-A-0026, WS-1358A, Ro-25-1132, CG|B- of one or more non-ligand components for binding (e.g., 57701,CGP -37697A, TMA-230, Syn-2190, , CS-834, DWB-204, DX-8739, CS-976, CKD-529, Ca", Mg" or a water molecule is required for the binding ER-35786, DZ-2640, 4-AAZ, KR-21012, RO-25-0993, of a ligand to various ligand binding sites). Examples. of DA-1211, BMS-181139, J-11225, L-786392, DK-35C, ligands useful in this invention are described herein. Those Ro-25-6833, S-1090, E-1 101, FK-518, DP-736, , skilled in the art will appreciate that portions of the ligand LY-215891, R0-09-1428, , , KST structure that are not essential for specific molecular recog 150185, Ro-09-1227, , , , nition and binding activity may be varied Substantially, LB-10522, Cefcanel, BRL-57342, Ce?prirome, YH-1226, replaced or substituted with unrelated structures (for , CKD-604, KST-150288, , Ro-24-8138, example... with ancillary groups as defined below) and, in FK-312, , RU-59863, , FR-193879, Some cases, omitted entirely without affecting the binding FK-041, , CP-6679, R0-63-9141, CFC-240, Cefpi interaction. The primary requirement for a ligand is that it mizole, Cefnminox, , CP-0467, PGE-7119699, has a ligand domain as defined above. It is understood that RO-48-8391, AM-1817, AM-1732, MC-02002, BO-1341, the term ligand is not intended to be limited to compounds BK-218, Ro-25-4835, RO-25-2016, YM-40220, Ro-23 known to be usefull in binding to penicillin binding proteins, 9424, LY-206763, CR-240, YH-1266, MC-02331, Ro-44 a pencillinase enzyme, a cephalosporinase enzyme, a 3949, MC-02306, Ro-25-7103, BMS-180680. Preferred transpeptidase enzyme, a Substrate of a transpeptidase B-lactam antibiotics are Amoxicillin, Nafcillin, Cefadroxil, enzyme, a beta-lactamase enzyme, a transglycoslase Ceftriaxone, Cefaclor, Aztreonam, Ceftazidime, , enzyme, or a transglycosylase enzyme Substrate (e.g., Meropenem, Ritipenem, Ceftazidine, Pipericillin, Clauv known drugs). Those skilled in the art will understand that linic acid, Cefepime, Cefoxitin, Cefotaximne, , the term ligand can equally apply to a molecule that is not Lefluzidine and derivatives thereof normally associated with penicillin binding proteins, a trans 0.175. The glycopeptide antibiotics are characterized by a glycoslase enzyme, or a taansglycosylase enzyme substrate multi-ring peptide core and at least one Sugar attached at binding properties. In addition, it should be noted that various sites, of which Vancomycin is an important example. ligands that exhibit marginal activity or lack useful activity Examples of the glycopeptide class of ligands included in as monomers can be highly active as multivalent compounds this definition may be found in "Glycopeptides Classifica because of the benefits conferred by multivalency. The term tion, Occurrence, and Discovery” by Rao, R. C. and Cran “ligand’ or “ligands' as used herein is intended to include dall, L. W., (Drugs and the Pharmaceutical Sciences' Vol. the racemic forms of the ligands as well as individual 63, edited by Ramakrishnan Nagarajan, published by Marcal enantiomers and diasteromers and non-racemic mixtures Dekker, Inc.) which is hereby incorporated by reference. thereof. Disclosed are glycopeptides identified as Actaplanin, Acti nodidin, Ardacin, Avoparcin, AZureomycm, A477, A35512, 0173 The term “B-lactamantibiotic” refers to antibiotics, A40926, A41030, A42867, A47934, A80407, A82846, having a B-lactam ring core which can be depicted as A83850, A84575, A84428, AB-65, Balhimycin, Chloroer follows: emomycin, Chloroorientiein, Chloropolysporin, Decapla US 2007/0134729 A1 Jun. 14, 2007

nin, N-demethylvancomycin. Eremomycin, Galacardin, Chem. Soc., Perkin Trans. I (1989), (12), 2335-9, and “The Helvecardin. IZupeptin, Kibdelin, LL-AM374, Mannopep Edman degradation of Vancomycin:preparation of Vancomy tin, MM45289, MM47756, MM47761, MM47921, cin hexapeptide', Booth, P. M., Stone, D.J. M. Williams, D. MM47766, MM55260, MM55266, MM55270, MM56579, H., Univ. Chem. Lab., Cambridge, UK., J. Chem. Soc., MM56598, OA-7653, Oreenticin, Parvodicin. Ristocetin, Chem. Commun. (1987), (22), 1694-5. Ristomycin, Synrmonicin, Teicoplanin, UK-68597, 0.178 "Optionally substituted Vancomycin with respect UK-69542. UK-72051, Vancomycin, and the like. Another to the multibinding agents of the invention refers to Vanco preferred class of ligands is the general class of glycopep mycin in which the hydroxy group at any position, the R tides disclosed above on which the Sugar moiety is absent. position the carboxyl groups at the C position, or the amine For example removal of the disaccharide moiety appended groups at the Vor N position that are not attached to the to the phenol on Vancomycin (as shown below as Formula linker X may or may not be substituted by various groups. II) by mild hydrolysis gives Vancomycin aglycone. A further Such groups include: R' where R at each occurrence is preferred class are glycopeptides that have been further chosen from alkyl, alkyl optionally interrupted by 1-5 appended with additional saccharide residues...especially atoms-chosen from O, S, or NR where R is alkyl, aryl, aminoglycosides, in a manner similar to Vancosamine. or heteroaryl, all of which are optionally. Substituted, 0176) “Vancomycin refers to the antibacterial compound haloalkyl, alkenyl, alkynyl, alkylamino, alkylaminoalkyl, whose structure is reproduced below as Formula Ia. cycloalkyl, alkanoyl, aryl, heteroaryl, heterocyclic, addi

Formula Ia

R

0177) “Optional” or “optionally” means that the subse tional saccharide residues, especially ammoglycosides, all quently described event or circumstance may or may not of which are optionally substituted as defined above; and: occur, and that the description includes instances where said NRWd in which R and R are independently hydrogen, event or circumstance occurs and instances in which it does alkyl, alkenyl, alkynyl, cycloalkyl, alkanoyl, aryl, het not. For example. "Optionally substituted glycopeptide' eroaryl, arylalkyl, or heteroarylalkyl, or R and R' when with respect to a compound of Formula (I) refers to a ligand taken together with the nitrogen atom to which they are as defined above in which those positions that are not linked to X may or may not be substituted by various groups as attached represent a heterocyclic group, quarternary alkyl defined below. The term also includes those instances in and aryl ammonium compounds, pyidinium ions, Sulfonium which one amino acid of the basic core structure is replaced ions, and the like, all of which are optionally substituted as by another amino acid, for example as described in “Prepa defined above. An example of a preferred C substitution is ration and conformational analysis of Vancomycin hexapep dimethylaminopropylamino and glucosamino; and example tide and aglucovancomycin hexapeptide', by Booth, P. M., of a preferred V substitution is alkyl, for example n-decyl, Williams, D. H., Univ. Chem. Lab., Cambridge, UK., J. or alkylaminoalkyl, for example n-decylaminoethyl. US 2007/0134729 A1 Jun. 14, 2007

0179 “Optionally substituted Vancomycin aglycone' with respect to the multibinding agents of the invention refers to Vancomycin aglycone in which the hydroxy group at any position, particularly the hydroxy group at the O position, the R position, the carboxy groups at the C position, or the amine group at the N position, that are not attached to the linker X may or may not be substituted by various groups —R" as defined above. 0180 “Transglycosylase enzyme Substrate” as used herein denotes the molecular target of the transglycosylase when only one ligand is interacting with a ligand binding enzyme. The substrate binds to the enzyme and eventually site. Examples of univalent interactions are depicted below. results in the synthesis of the bacterial cell wall. The action 0183 The term “multivalency” as used herein refers to of this enzyme is inhibited by a ligand domain that binds to the concurrent binding of from 2 to 10 linked ligands (which the enzyme itself and/or the enzyme Substrate. A ligand Such may be the same or different) and two or more correspond as Vancomycin binds to this Substrate and in effect “seques ing ligand binding sites which May be the same or different. ters' the substrate to prevent its recognition by the enzyme 0.184 For example, two ligands connected through a and Subsequent use in the construction of the bacterial cell linker that bind concurrently to two ligand binding sites wall. There is also a growing feeling that some glycopep would be considered as bivalency; three ligands thus con tides or derivatives thereof may directly bind to and inhibit nected would be an example of trivalency. An example of the transglycolase. trivalent binding, illustrating a multibinding compound bearing three ligands versus a monovalent binding interac 0181. The term “multibinding compound or agent” refers tion, is shown below: to a compound that is capable of multivalency, as defined below, and which has 2-10 ligands covalently bound to one or more linkers. In all cases, each ligand and linker in the multibinding compound is independently selected Such that ((- - - (e. the multibinding compound includes both symmetric com univalent interaction pounds (i.e., where each ligand as well as each linker is identical) and asymmetric compounds (i.e., where at least one of the ligands is different from the other ligand(s) and/or s at least one linker is different from the other linker(s)). Multibinding compounds provide a biological and/or thera peutic effect greater than the aggregate of unlinked ligands trivalent interaction equivalent thereto which are made available for binding. That is to say that the biological and/or therapeutic effect of the ligands attached to the multibinding compound is greater 0185. It should be understood that not all compounds that contain multiple copies of a ligand attached to a linker or to than that achieved by the same amount of unlinked ligands linkers necessarily exhibit the phenomena of multivalency, made available for binding to the ligand binding sites i.e., that the biological and/or therapeutic effect of the (receptors). The phrase “increased biological or therapeutic multibinding agent is greater than the sum of the aggregate effect” includes, for example: increased affinity, increased of unlinked ligands made available for binding to the ligand selectivity for target, increased specificity for target, binding site (receptor). For multivalency to occur, the increased potency, increased efficacy, decreased toxicity, ligands that are connected by a linker or linkers have to be improved duration of activity or action, increased ability to presented to their ligand binding sites by the linker(s) in a kill cells Such as fungal pathogens, cancer cells, etc., specific manner in order to bring about the desired ligand decreased side effects, increased therapeutic index, orienting result, and thus produce a multibinding event. improved bioavailibity, improved pharmacokinetics, 0186 The term “potency” refers to the minimum con improved activity spectrum, and the like. The multibinding centration at which a ligand is able to achieve a desirable biological or therapeutic effect. The potency of a ligand is compounds of this invention will exhibit at least one and typically proportional to its affinity for its ligand binding preferably more than one of the above-mentioned affects. site. In some cases, the potency may be non-linearly corre lated with its affinity. In comparing the potency of two drugs, 0182. The term “univalency” as used herein refers to a e.g., a multibinding agent and the aggregate of its unlinked single binding interaction between one ligand as defined ligand, the dose-response curve of each is determined under herein with one ligand binding site as defined herein. It identical test conditions (e.g., in an in vitro or in vivo assay, should be noted that a compound having multiple copies of in an appropriate animal model). The finding that the multi a ligand (or ligands) exhibit univalency binding agent produces an equivalent biological or thera US 2007/0134729 A1 Jun. 14, 2007 peutic effect at a lower concentration than the aggregate 0197) The term “therapeutically effective amount” refers unlinked ligand is indicative of enhanced potency. to that amount of multibinding compound which is sufficient to effect treatment, as defined above, when administered to 0187. The term “selectivity” or “specificity” is a measure a mammal in need of Such treatment. The therapeutically of the binding preferences of a ligand for different ligand effective amount will vary depending upon the Subject and binding sites (receptors). The selectivity of a ligand with disease condition being treated, the weight and age of the respect to its target ligand binding site relative to another Subject, the severity of the disease condition, the manner of ligand binding site is given by the ratio of the respective administration and the like, which can readily be determined values of K (i.e., the dissociation constants for each ligand by one of ordinary skill in the art. receptor complex) or, in cases where a biological effect is observed below the Y, the ratio of the respective ECso's 0198 The term “linker, identified where appropriate by (i.e., the concentrations that produce 50% of the maximum the symbol X refers to a group or groups that covalently response for the ligand interacting with the two distinct attaches from 2 to 10 ligands (as identified above) in a ligand binding sites (receptors)). manner that provides for a compound capable of multiva lency. Among other features, the linker is a ligand-orienting 0188 The term “ligand binding site' denotes the site on entity that permits attachment of at least two copies of a a penicillin binding proteins, a transpeptidase enzyme, peni ligand (which may be the same or different) thereto. In some cillinase enzyme, cephalosporinase enzyme, beta lactamase cases, the linker may itself be biologically active. The term enzyme, a transpeptidase enzyme Substrate, a transglycosy “linker” does not, however, extend to cover solid inert lase enzyme and/or transglycosylase enzyme Substrate that Supports Such as beads, glass particles, fibers, and the like. recognizes a ligand domain and provides a binding partner But it is understood that the multibinding compounds of this for the ligand. The ligand binding site may be defined by invention can be attached to a solid support if desired. For monomeric or multimeric structures. This interaction may example, Such attachment to Solid Supports can be made for be capable of producing a unique biological effect, for use in separation and purification processes and similar example, agonism, antagonism, and modulatory effects, or it applications. may maintain an ongoing biological event, and the like. 0199 The extent to which multivalent binding is realized 0189 It should be recognized that the ligand binding sites depends upon the efficiency with which the linker or linkers of the enzyme or the receptor that participate in biological that joins the ligands presents these ligands to the array of multivalent binding interactions are constrained to varying available ligand binding sites. Beyond presenting these degrees by their intra- and inter-molecular associations. For ligands for multivalent interactions with ligand binding example, ligand binding sites may be covalently joined to a sites, the linker or linkers spatially constrains these interac single structure, noncovalently associated in a multimeric tions to occur within dimensions defined by the linker or structure, embedded in a membrane or polymeric matrix, linkers. Thus, the structural features of the linker (valency, and so on and therefore have less translational and rotational geometry, orientation, size, flexibility, chemical composi freedom than if the same structures were present as mono tion, etc.) are features of multibinding agents that play an mers in Solution. important role in determining their activities. 0190. The term “inert organic solvent” or “inert solvent” 0200. The linkers used in this invention are selected to means a solvent, which is inert under the conditions of the allow multivalent binding of ligands to the ligand binding reaction being described in conjunction therewith including, sites of an enzyme involved in cell wall biosynthesis and by way of example only, benzene, toluene, acetonitrile, metabolism, a precursor used in the synthesis of the bacterial tetrahydrofuran, dimethylformamide, chloroform, methyl cell wall and/or the cell surface, whether such sites are ene chloride, diethyl ether, ethyl acetate, acetone, methyl located interiorly, both interiorly and on the periphery of the ethyl ketone, methanol, ethanol, propanol, isopropanol, t-bu enzyme structure, or at any intermediate position thereof. tanol, dioxane, pyridine, and the like. Unless specified to the 0201 In the FIGS. 9, 10, 14-16, glycopeptides are contrary, the solvents used in the reactions described herein depicted in a simplified form as a shaded box that shows are inert solvents. only the carboxy terminus, labeled C), the Sugar amine 0191 The term “treatment” refers to any treatment of a terminus (e.g., Vancosamine), labeled IV), and the “non pathologic condition in a mammal, particularly a human, sugar amino terminus, labeled N as follows: and includes: 0192 (i) preventing the pathologic condition from occur ring in a Subject which may be predisposed to the condition H O but has not yet been diagnosed with the condition and, N accordingly, the treatment constitutes prophylactic treatment R1 O for the disease condition; N C 0193 (ii) inhibiting the pathologic condition, i.e., arrest V ing its development; 0194 (iii) relieving the pathologic condition, i.e., causing regression of the pathologic condition; or where R is hydrogen (as N-desmethylvancomycin) or 0.195 (iv) relieving the conditions mediated by the patho methyl (as in Vancomycin). logic condition. 0202) It can be seen by way of exemplification that one 0196. The term “pathologic condition which is modu class of multivalent compounds that fall within the scope of lated by treatment with a ligand’ covers all disease states (i.e., pathologic conditions) which are generally acknowl the definition of Formula (I) include compounds wherein the edged in the art to be usefully treated with a ligand that is an glycopeptide ligand is connected by one or more linkers at antibacterial agent, and those disease states which have been the C. V. or N terminus. found to be usefully treated by a specific multibinding 0203 Another class of multivalent compounds that fall compound of our invention. within the scope of the definition of Formula (I) include US 2007/0134729 A1 Jun. 14, 2007

compounds where the aglycone derivatives of glycopep wherein: tides. depicted as a triangle that shows only the carboxyl terminus, labeled C, the aglycone hydroxy terminus 0208 R is substituted alkyl, aryl, aralkyl, or heteroaryl labeled 9), and the “non-sugar amino terminus, labeled N) wherein each of said Substituent optionally links (a) to a as follows: linker via a covalent bond or R is a covalent bond that links OH (a) to a linker; and O 0209) R' and Rare, independently of each other, alkyl or at least one of R' and R is a covalent bond linking (a) to a linker; 0210 (ii) a compound of formula (b): where R is hydrogen (as in N-desmethylvancomycin agly (b) cone) or methyl (as in Vancomycin aglycone) wherein the R5 aglycone derivatives ligand is connected by one or more Q linkers at the C. V. or N terminus. R3-CO-NH Np 0204 Athird class of compounds falling within the scope N 21 R4 of the invention are those in which the glycopeptides, or O aglycone derivatives thereof, are linked via the R position. Reaction Schemes that exemplify this linking strategy depict COOH the ligands in a simplified form as above, i.e., as a shaded box in which the carboxyl terminus is labeled C), the Vancosamine amino terminus is labeled IV), and the “non wherein: sugar amino terminus is labeled N), with the addition of the R position as a resorcinol derivative as shown below: 0211 one of P and Q is O, S, or —CH2— and the other O is —CH2—, 0212 R is substituted alkyl, heteroarylalkyl, aralkyl, N R1 OH heterocyclylalkyl, or - C)=NOR7 (where R is aryl, C heteroaryl, or substituted alkyl; and R is alkyl or substituted alkyl) wherein each of said substituent optionally links (b) to HO VNH a linker or R is a covalent bond that links (b) to a linker; and 0213) R' is hydrogen, alkyl, alkenyl, substituted alk enylene, Substituted alkyl, halo, heteroarylalkyl, heterocy R clylalkyl, - SR" (where R is aryl, heteroaryl, heterocyclyl, OH or cycloalkyl) or —CHSR' (where R is aryl, heteroaryl, heterocyclyl, or cycloalkyl) wherein each of said substituent where R is hydrogen or methyl. optionally links (b) to a linker or R' is a covalent bond that links (b) to a linker; PREFERRED EMBODIMENTS 0214) R is hydrogen, hydroxy, or alkoxy; 0205 While the broadest definition of this invention is set forth in the Summary of the Invention, certain compounds of 0215 (iii) a compound of formula (c): Formula (I) are preferred. (A) One preferred group of compounds is a multibinding compound of Formula (II): L X L (II) wherein: 0206 L' is a beta lactam antibiotic is selected from the group consisting of 0207 (i) a compound of formula (a):

wherein: (a) O 0216) T is S or CH:

- NH 0217 R is alkyl; 0218 W is O, S, OCH , or CH; and R is -(alky lene)-NHC(Rb=NH where R is a covalent bond linking (c) COOH to a linker; or - W R is a covalent bond that links (c) to a linker, US 2007/0134729 A1 Jun. 14, 2007

0219 (iv) a compound of formula (d): 0228 (i) a compound of formula (a):

(a) O (d)

- NH S RI R2

N O COOH

wherein: wherein: 0229) R is: 0220 R and R are alkyl;

7 o 7 o 0221) R' is selected from the group consisting of hydro R YC R YC gen, alkyl, Substituted alkyl, halo, aryl, heteroaryl, hetero cyclyl, aralkyl, heteroaralkyl, heterocyclylalkyl or \ / CH- \ / OCH —CHSR' (where R' is aryl, heteroaryl, heterocyclyl, or OCH cycloalkyl) wherein each of said substituent optionally links (d) to a linker or at least one of RandR' is a covalent bond R3 o that links (d) to a linker; or SC ro-()-- \ / NHRloss

0222 RandR' together with the carbonatoms to which they are attached form an aryl, heteroaryl, cycloalkyl, Sub stituted cycloalkyl, or heterocyclyl ring of 4 to 7 ring atoms wherein one of the ring atoms optionally links (d) to a linker; O 0223 (v) a compound of formula (e):

OCH5 R20 = R2 = H R20 = R2 = Cl (e) R20 = Cl and R2 = H RIS o YC CH- Cy CH \ / S COORI9 COOR17 R3SC o

\ / NHCO V wherein: 0224) R' is -SOH or -(alkylene)-COOH: S- O C V 0225) R' is alkyl, substituted alkyl, haloalkyl, alkoxy, SOCH3 aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, Substi R. o tuted cycloalkyl, or heterocyclyl wherein each of said sub YC stituent optionally binds (e) to a linker or R'' is a covalent N / NHCO bond that links (e) to a linker; and V 0226) R' is alkyl, acyl, or COC(R)=N OR' wherein R'' is aryl, heteroaryl which optionally links (e) to a linker, and R' is -(alkylene)-COOR' wherein R' is CCO hydrogen or optionally links (e) to a linker or R' is a covalent bond that links (e) to a linker, preferably 0227 L' is selected from the group consisting of. US 2007/0134729 A1 Jun. 14, 2007 19 where: 0233 (ii) a compound of formula (b): (b) 0230) R'' is a covalent bond that links the (a) group to a linker; R3-CO-NH

0231 one of R' and R' is hydrogen and the other is a covalent bond that links the (a) group to a linker, and where: 0232) R' and k are methyl: 0234 Rand Rare:

R3 R 4 RN/ffly-a- - CHOCOCH S

NN-CH,- N ic-C-en

R18SC —CH

NHR19 ro-( )-i- —CH

SCH SCH NS NS ? O / Ns N -CH3 N - N-CHSO N N CHOCONHR'. CHOCOCH,

NHR19

R18 t CHOCONHR'

O NOCH

RISSC SCH y \ CH2 o N / N N-CHCOOH

US 2007/0134729 A1 Jun. 14, 2007

wherein: -continued 0235 n is 0 or 1; mis 1-5; Z is CH or N: Y is H or halo: R is alkyl: R'' is a covalent bond that links the (b) group to a linker; one of R' and R' is hydrogen or alkyl; R' and Rare, independently of each other, hydrogen or alkyl; or together with the nitrogen atom to which they are attached form a heterocycloamino group; and R, R and R are independently alkyl wherein one of R. R. R. R. is a covalent bond that links the (b) group to a linker; where: 0236 (iii) a compound of formula (c): 0238) R' is a covalent bond that links (d) to a linker; 0239 one of R and R is hydrogen, alkyl, substi (c ) tuted alkyl, or aralkyl, and other is a covalent bond that links (d) to a linker, R is alkyl; or sch.cit.NE-NH 0240 (v) a compound of formula (e): Rb

COOH

(e) wherein R is a covalent bond linking (c) to a linker; CH3 0237 (iv) a compound of formula (d): CH NEC (d)

SR

NHR22

wherein one of R and R’ is hydrogen and the other links (d) to a linker; 024.1 L is an optionally substituted Vancomycin which OH O is linked to a linker via any hydroxyl group, carboxyl group or amino group; and R25 0242 X is a linker and is selected from a compound of O-tyN ty--- formula: N R26 k NR25 R24 wherein ty-or 0243 m is an integer of from 0 to 20; N 0244 X at each separate occurrence is selected from the group consisting of —O— —S— —NR—, —C(O)—, R23 C(O)O , OC(O) , C(O)NR-, - NRC(O) , O C(S), —C(S)O , —C(S)NR— —NRC(S)—, or a covalent O bond where R is as defined below: 0245 Z at each separate occurrence is selected from the s l group consisting of alkylene, Substituted alkylene, N- N R25 cycloalkylene, Substituted cylcoalkylene, alkenylene, Sub R24 stituted alkenylene, alkynylene, Substituted alkynylene, R25 cycloalkenylene, Substituted cycloalkenylene, arylene, het R25 N eroarylene, heterocyclene, or a covalent bond; NHSO1 W 0246 each ya at each separate occurrence is selected N from the group consisting of —O—, —C(O)—, OC(O) , C(O)O , NR S(O)n , R24 C(O)NR' NR'C(O) , NRC(O)NR' , R24 NR'C(S)NR' , C(=NR). NR , NR C(=NR) , OC(O) NR' , NR' C(O)-O-, US 2007/0134729 A1 Jun. 14, 2007 22

N=C(X) NR NR C(XY N , alkyl) wherein each of said substituient optionally links (b) P(O)(OR") O - O P(O)(OR") , S(O)CR'R" , to a linker or R is a covalent bond that links (b) to a linker; S(O), NR' —NR' S(O), . —S S , and a cova and lent bond; where n is 0, 1 or 2; and R, R and R'' at each separate occurrence are selected from the group consisting 0254) R' is hydrogen, alkyl, alkenyl, substituted alk of hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted, enylene, Substituted alkyl, halo, heteroarylalkyl, heterocy cycloalkyl, alkenyl, Substituted alkenyl, cycloalkenyl, Sub clylalkyl, - SR" (where R is aryl, heteroaryl, heterocyclyl, stituted cycloalkenyl, alkynyl, Substituted alkynyl, aryl, het or cycloalkyl) or —CHSR' (where R is aryl, heteroaryl, eroaryl and heterocyclic; and pharmaceutically acceptable heterocyclyl, or cycloalkyl) wherein each of said substituent salts thereof provided that when L is vancomycin attached optionally links (b) to a linker or R' is a covalent bond that to a linker via the C terminus, then L cannot be cefalexin links (b) to a linker; attached to the linker via acylation of its alpha amino group; and pharmaceutically acceptable salts thereof. 0255 R is hydrogen, hydroxy, or alkoxy; (B) Another more preferred group of compounds is a multi 0256 (iii) a compound of formula (c): binding compound of Formula (III): L X L (III)

(c) wherein: 0247 ligands, L and L', are a beta lactam antibiotic and are independently selected from the group consisting of: 0248 (i) a compound of formula (a):

(a) O wherein:

- NH RI 0257) T is S or CH: 0258 R is alkyl; 0259 W is O, S, OCH , or CH; and R is -(alky O COOH lene)-NHC(R)=NH where R is a covalent bond linking (c) to a linker; or - W R is a covalent bond that links (c) to a linker; wherein: 0260 (iv) a compound of formula (d):. 0249 R is substituted alkyl, aryl, aralkyl, or heteroaryl wherein each of said Substituent optionally. links (a) to a linker via a covalent bond or R is a covalent bond that links (d) (a) to a linker, and 0250) R' and Rare, independently of each other, alkyl or at least one of R' and R is a covalent bond linking (a) to a linker; 0251 (ii) a compound of formula (b):

(b) wherein: R5 R3-CO-NH Q Np 0261) R' and R* are alkyl:

N 0262) R' is selected from the group consisting of hydro 21 R4 gen, alkyl, Substituted alkyl, halo, aryl, heteroaryl, hetero O cyclyl, aralkyl, heteroaralkyl, heterocyclylalkyl or COOH —CHSR' (where R is aryl, heteroaryl, heterocyclyl, or cycloalkyl) wherein each of said substituent optionally links wherein: (d) to a linker or at least one of RandR' is a covalent bond that links (d) to a linker; or 0252 one of P and Q is O, S, or —CH2— and the other 0263. RandR' together with the carbonatoms to which is —CH2—, they are attached form an aryl, heteroaryl, cycloalkyl, Sub 0253) R is substituted alkyl, heteroarylalkyl, aralkyl, stituted cycloalkyl, or heterocyclyl ring of 4 to 7 ring atoms heterocyclylalkyl, or - C )=NOR7 (where R is aryl, wherein one of the ring atoms optionally links (d) to a linker; heteroaryl, or substituted alkyl; and R is alkyl or substituted O US 2007/0134729 A1 Jun. 14, 2007

0264 (v) a compound of formula (e): -continued OCH R3YC o (e) \ / R19 O ti NHR18 OCH

R13 {?/ \ wherein:

0265 R'' is -SOH or -(alkylene)-COOH: R20 = R2 = H OCHs R20 = R2 = Cl 0266) R' is alkyl, substituted alkyl, haloalkyl, alkoxy, R20 = Cl and R2 = H aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, Substi tuted cycloalkyl, or heterocyclyl wherein each of said sub RIS o stituent optionally binds (e) to a linker or R' is a covalent YC CH- CH \ / S bond that links (e) to a linker; and COORI9 COOR17

0267) R' is alkyl, acyl, or COC(R)=N OR'S 7 o wherein R'' is aryl, heteroaryl which optionally links (e) to RSC a linker, and R' is -(alkylene)-COOR' wherein R' is N / NHCO hydrogen or optionally links (e) to a linker or R' is a V covalent bond that links (e) to a linker, preferably S- O 0268 L and L are independently selected from the C V group consisting of SOCH 0269 (i) a compound of formula (a): RSC7 o \ y NHCO (a) V O ul CH R NH S 3 CH3 CCO

N CH5 O COOH where: 0271) R' is a covalent bond that. links the (a) group to a linker, one of R' and R' is hydrogen and the other is a wherein: covalent bond that links the (a) group to a linker, 0270 R is: 0272 (ii) a compound of formula (b):

(b) R3-CO-NH

US 2007/0134729 A1 Jun. 14, 2007 25

-continued R3 R? Y R19 S \ SAS

R18NH -S Nt 2

19 NS-R'O HC Y R R S "NQ \ R NR), RINH18 -SN N No1 R19 R SCH O CH- N / NHCO NSN R N

CCO C2H5 Y 21 NZ -SS \ Ns S R18NH N N No1 R19 Y R30 R31 Y--1N SN3Y(R32), 21 S 1N R33

(Note: the R group in the left column is paired with the R' in the right column) wherein: 0276 (iii) a compound of formula (c):

0274) n is 0 or 1; m is 1-5; Z is CH or N: Y is H or halo: R is alkyl: (c)

0275 R7 is a covalent bond that links the (b) group to a linker; one of R'' and R' is hydrogen or alkyl; Rand R' are, independently of each other, hydrogen or alkyl; or together with the nitrogen atom to which they are attached form a heterocycloamino group; and R, R and R are independently alkyl wherein one of R. R. R. R. is a 0277 wherein R is a covalent bond linking (c) to a covalent bond that links the (b) group to a linker; linker; US 2007/0134729 A1 Jun. 14, 2007 26

0278 (iv) a compound of (c) formula (d): 0281 (v) a compound of formula (e):

(d) (e)

SRa CH3

CH3 NEC

where R is: YNHR22

OH O R25 wherein one of R and R’ is hydrogen and the other links O-ty ty--- (d) to a linker, and N R26 24 0282 X is a linker is selected from a compound of R Yes R24 formula:

ty-or wherein N 0283 m is an integer of from 0 to 20; R23 0284 X at each separate occurrence is selected from the O group consisting of —O— —S— —NR—, —C(O)—, C(O)O , OC(O) , C(O)NR-, - NRC(O) , O C(S), —C(S)O , —C(S)NR— —NRC(S)—, or a covalent N-r ls R25 bond where R is as defined below: R24 0285 Z at each separate occurrence is selected from the group consisting of alkylene, Substituted alkylene, R25 cycloalkylene, Substituted cylcoallylene, alkenylene, Substi R25 N tuted alkenylene, alkynylene, Substituted alkynylene, NHSO1 W cycloalkenylene, Substituted cycloalkenylene, arylene, het N eroarylene, heterocyclene, or a covalent bond; R24 0286 each Y at each separate occurrence is selected R24 from the group consisting of —O—, —C(O)—, OC(O) , C(O)O , NR S(O)n , O C(O)NR' , NRC(O), NRC(O)NR' , NR'C(S)NR' , C(=NR). NR , NR N(CH3)2 C(=NR) , OC(O) NR' , NR' C(O)-O-, N N=C(X) NR , NR C(X)=N , R23 - P(O)(OR) O , - O -P(O)(OR") — S(O)CR'R" , S(O) NR' , – NR S(O), , —S–S , and a covalent bond; where n is 0, 1 or 2; and R, R" and R' at each separate occurrence are selected from the where: group consisting of hydrogen, alkyl, Substituted alkyl, 0279 R is a covalent bond that liniks (d) to a linker; cycloalkyl, Substituted cycloalkyl, alkenyl, Substituted alk 0280 one of R and R is hydrogen, alkyl, substi enyl, cycloalkenyl, Substituted cycloalkenyl, alkynyl. Sub tuted alkyl, or aralkyl, and other is a covalent bond that stituted alkynyl, aryl, heteroaryland heterocyclic; and phar links (d) to a linker; R is alkyl; or maceutically acceptable salts thereof. US 2007/0134729 A1 Jun. 14, 2007 27

0287. Within the above more preferred groups, an even more preferred group of compounds is that wherein: 0288) L, L, and L are independently selected from the group consisting of

US 2007/0134729 A1 Jun. 14, 2007 28

-continued

NHMe US 2007/0134729 A1 Jun. 14, 2007 29

-continued O HO N-1a1n-1-1-1N1-S-1a H

NHMe and

\ HN-H

O

n1n N1 H v

NHMe wherein the atom carrying the bond with the dashed line indicates the point of attachment of the ligand to the linker; -continued and

0289 the linker is selected from the group consisting of: US 2007/0134729 A1 Jun. 14, 2007 30

-continued -continued

-V HN Y -V HN

N H s

X w

T NH a X

NHX a

US 2007/0134729 A1 Jun. 14, 2007 31

-continued -continued

chiral

chiral

N-CH

a 4. A. a

8 A. / K HC-N

O

N-CH x s a x 8 w 8 US 2007/0134729 A1 Jun. 14, 2007 32

-continued -continued

US 2007/0134729 A1 Jun. 14, 2007 33

-continued -continued

HN

O

NH2

'ss O NH

US 2007/0134729 A1 Jun. 14, 2007 34

-continued -continued

w O w F w p w P w F - OH

w HN O NH w 'nu-1S-1N1-1N1 ? A w H ?

w w w F

HN N H HN A.

OH

LINKERS DERIVED FROM AMINOALDEHYDES A~~~~ Ny

^-n-SS US 2007/0134729 A1 Jun. 14, 2007

-continued -continued O Ny O H

NH ? ', ?

O F O '', N.' 0290 Representative compounds of the invention are , ? shown in the table below:

N 0291 (I) Compounds of Formula (III) wherein the ? ligands are selected from a compound of formula (b) and are O y linked to a linker, X. via the R group and where P, Q. R. and R and are as defined below are:

COOH O R4

P 5 s 1. -Q S NHCO-X-CONE R Q P

N 4 R O COOH

Cpd No Linker X

US 2007/0134729 A1 Jun. 14, 2007 36

-continued

COOH

O N 1. R4 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O

US 2007/0134729 A1 Jun. 14, 2007 37

-continued

COOH

O N 1. R4 R5 o1 P -Q S NHCO-X-CONH R5 P -

N O COOH

N US 2007/0134729 A1 Jun. 14, 2007 38

-continued

COOH

O N 1. R4 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O

10

11

12

US 2007/0134729 A1 Jun. 14, 2007 39

-continued

COOH

O N 1. R4 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O

13

14

US 2007/0134729 A1 Jun. 14, 2007 40

-continued

COOH

O N 1. R4 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O

15

16 US 2007/0134729 A1 Jun. 14, 2007 41

-continued

O N 1 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O COOH cro r O s CC CC - - 18 SOro, O O r O s CC CC - - 19 s H NH SO O O O CC CC - - US 2007/0134729 A1 Jun. 14, 2007 42

-continued

COOH

O N 1. R4 R5 o1 P 1Q S NHCO-X-CONH R5 P

N O

21

US 2007/0134729 A1 Jun. 14, 2007 44

-continued

COOH

s P R5 NHCO-X-CONHS o1 1. Q a.S R5

P

4 R O COOH

21 —CH2-1-methyl- H -CH2- S 1H-tetrazol-5- ylsulfanyl 22 -CH2-1-methyl- H -CH2- S 1H-tetrazol-5- ylsulfanyl

0292 Compounds of Formula (III) wherein the ligands are selected from a compound of formula (1) and are linked to a linker, X, via the R group and where P, Q, R, and R' and are as defined below are:

5 5 RCONH R Q Q R SNHCOR P P

N 2 X S N O O COOH COOH

Cpd No Linker X P Q R3 R 1 --- O (2-aminothiazol-4-yl)-methoxyiminomethyl H -CH2— S O l N N

21 N US 2007/0134729 A1 Jun. 14, 2007 45

III. Other compounds of the invention are:

HO O

O NN. 21 N NH O

2 S N N=( CH O H S

N - \N N N -N | N O s1N O CH /

- O y \ N S Nu S N H N s1 O-N

)= O DC 21 O CH / HN O N P- NSSa US 2007/0134729 A1 Jun. 14, 2007 46

-continued

General Synthetic Scheme like. Such materials may be characterized using conven 0293 Compounds of this invention can be made by the tional means, including physical constants and spectral data. methods depicted in the reaction schemes shown below. 0296. Furthermore, it will be appreciated that where 0294 The starting materials and reagents used in prepar typical or preferred process conditions (i.e., reaction tem ing these compounds are either available from commercial peratures, times, mole ratios of reactants, solvents, pres suppliers such as Aldrich Chemical Co., (Milwaukee, Wis., Sures, etc.) are given, other process conditions can also be USA), Bachem (Torrance, Calif., USA), Emka-Chemie, or used unless otherwise stated. Optimum- reaction conditions Sigma (St. Louis, Mo., USA) or are prepared by methods may vary with the particular reactants or solvent used, but known to those skilled in the art following procedures set such conditions can be determined by one skilled in the art forth in references such as Fieser and Fieser’s Reagents for by routine optimization procedures. Organic Synthesis, Volumes 1-15 (John Wiley and Sons, 0297 Additionally, as will be apparent to those skilled in 1991); Rodd's Chemistry of Carbon Compounds, Volumes the art, conventional protecting groups may be necessary to 1-5 and Supplementals (Elsevier Science Publishers, 1989), prevent certain functional groups from undergoing undes Organic-Reactions, Volumes 140 (John Wiley and Sons, ired reactions. The choice of a Suitable protecting group for 1991), March's Advanced Organic Chemistry, (John Wiley a particular functional group as well as Suitable conditions and Sons, 4th Edition), and Larock’s Comprehensive for protection and deprotection are well known in the art. Organic Transformations (VCH Publishers Inc., 1989). For example, numerous protecting groups, and their intro 0295) The starting materials and the intermediates of the duction and removal, are described in T. W. Greene and G. reaction may be isolated and purified if desired using M. Wuts, Protecting Groups in Organic Synthesis, Second conventional techniques, including but not limited to filtra Edition, Wiley, New York, 1991, and references cited tion, distillation, crystallization, chromatography, and the therein. US 2007/0134729 A1 Jun. 14, 2007 47

0298 These schemes are merely illustrative of some methods by which the compounds of this invention can be TABLE I synthesized, and various modifications to these schemes can be made and will be suggested to one skilled in the art Representative Complementary Binding Chemistries having referred to this disclosure. First Reactive Group Second Reactive Group Linkage carboxyl amine amide Sulfonyl halide amine Sulfonamide hydroxyl alkyl/aryl halide ether Preparation of a multibinding compound of hydroxyl isocyanate urethane Formlula (I) amine epoxide 3-hydroxyamine amine alkyl/aryl halide alkylamine 0299. In general, a multibinding compound of Formula hydroxyl carboxyl ester amine aldehyde/NaCNBH amine (I) can be prepared as illustrated and described in Scheme A hydroxylamine Sulfonyl halide Sulfonamide below. aldehyde amine/NaCHBH amine aldehyde amine/NaCHBH amine 0300. A multibinding compound of Formula (I) can be amine isocyanate 8 prepared by covalently attaching the ligands, L, to a linker, X, as shown in Scheme A below. 0304 By way of example, reaction between a carboxylic acid of either the linker or the B-lactam and a primary or secondary amine of the B-lactam or the linker in the presence Scheme A of Suitable, well-known activating agents such as dicyclo Method (a) hexylcarbodiimide, results in formation of an amide bond FG covalently linking the B-lactam to the linker, reaction 21 + FG2-X-FG2 - - L X L between an amine group of either the linker or the B-lactam Method (b) and a sulfonylbhalide of the B-lactarn or the linker, in the FG presence of a base Such as triethylamine, pyridine, an the L1 + FG2-X-FG2PG - - like results in formation of a sulfonamide bond covalently FG linking the B-lactam to the linker, and reaction between an d eprotect 2 Li 1 alcohol or phenol group of either the linker or the B-lactam L-X-FG2PG L-X-FG4 - > and an alkyl or aryl halide of the f-lactam or the B-lactarn intermediate in the presence of a base Such as triethylamine, pyridine, and (II) the like, results in formation of an ether bond covalently L-X-L linking the B-lactam to the linker. 0305 Any-compound which is an antibacterial agent can be used as a ligand in this invention. Typically, a compound 0301 In method (a), a multibinding compound of For selected for use as a ligand will have at least one functional mula (I) is prepared in one step, by covalently attaching the group, Such as an amino, hydroxyl, thiol or carboxyl group ligands, L, to a linker, X, where FG' and FG represent a and the like, which allows the compound to be readily functional group Such as halo, pseudohalides, boronates, coupled to the linker. Compounds having Such functionality amino, hydroxy, thio, aldehyde, ketone, carboxy, carboxy are either known in the art or can be prepared by routine derivatives such as acid halide, ester, amido, and the like. modification of known compounds using conventional This method is preferred for preparing compounds of For reagents and procedures. mula (I) where both the ligands are identical. 0306 Linkers can be attached to different positions on the ligand molecule to achieve different orientations of the 0302) In method (b), the compounds of Formula (I) are ligand domains, and thereby facilitate multivalency. While a prepared in a stepwise manner by covalently attaching one number of positions on the ligands are synthetically practical equivalent of a ligand, L, with a ligand X where FG' and for linking, it is:preferred to preserve those ligand Substruc FG' represent a functional group as defined above, and tures which are most important for ligand-receptor binding. FGPG is a protected finctional group to give an interme 0307. It will be apparent to one skilled in the art that the diate of formula (II). Deprotection of the second functional above chemistries are not limited to preparing multibind group on the ligand, followed by reaction with a ligand L. ingbinding compounds of Formula (I) and can be used to which may be same or different than ligand L, then pro prepare tri-, tetra-, etc., multibinding compounds of Formula vides a compound of Formula (I). This method is suitable for (I). preparing compounds of Formula (I) where the ligands are 0308 The linker is attached to the ligand at a position that the non-identical. retains ligand domain-ligand binding site interaction and specifically which permits the ligand domain of the ligand to 0303. The ligands are covalently attached to the linker orient itself to bind to the ligand binding site. Such positions using conventional chemical techniques providing for cova and synthetic protocols for linkage are well known in the art. lent linkage of the ligand to the linker. Reaction chemistries The term linker embraces everything that is not considered resulting in Such linkages are well known in the art and to be part of the ligand. involve the use of complementary functional groups on the 0309 The relative orientation in which the ligand linker and ligand as shown in Table I below. domains are displayed derives from the particular point or US 2007/0134729 A1 Jun. 14, 2007 48 points of attachment of the ligands to the linker, and on the ing diagram, although clearly the framework is a three framework geometry. The determination of where accept dimensional array in practice: able Substitutions can be made on a ligand is typically based on prior knowledge of structure-activity relationships (SAR) of the ligand and/or congeners and/or structural information about ligand-receptor complexes (e.g., X-ray crystallogra phy, NMR, and the like). Such positions and the synthetic methods for covalent attachment are well known in the art. Following attachment to the selected linker (or attachment to a significant portion of the linker, for example 2-10 atoms of the linker), the univalent linker-ligand conjugate may be 0 1 2 3 4 5 6 78 tested for retention of activity in the relevant assay. 0310. The linker, when covalently attached to multiple 0314. Each dot is either an atom, chosen from carbon, copies of the ligands, provides a biocompatible, Substan hydrogen, oxygen, nitrogen, Sulfur, phosphorus, or halogen, tially non-immunogenic multibinding compound. The bio logical activity of the multibinding compound is highly or the dot represents a point in space (i.e., an absence of an sensitive to the Valency, geometry, composition, size, flex atom). AS is apparent to the skilled artisan, only certain ibility or rigidity, etc. of the linker and, in turn, on the overall atoms on the grid have the ability to act as an attachment structure of the multibinding compound, as well as the point for the ligands, namely, C, O, N, S and P. presence or absence of anionic or cationic charge, the 0315 Atoms can be connected to each other via bonds relative hydrophobicity/hydrophilicity of the linker, and the (single, double or triple bonds with acceptable resonance like on the linker. Accordingly, the linker is preferably and tautomeric forms), with regard to the usual constraints chosen to maximize the biological activity of the multib of chemical bonding. Ligands may be attached to the frame inding compound. The linker may be chosen to enhance the work via single, double or triple bonds (with chemically biological activity of the molecule. In general, the linker acceptable tautomeric and resonance forms). Multiple ligand may be chosen from any organic molecule construct that groups (2 to 10) can be attached to the framework such that orients two or more ligands to their ligand binding sites to the minimal, shortest path distance between adjacent ligand permit multivalency. In this regard, the linker can be con groups does not exceed 100 atoms. Preferably, the linker sidered as a “framework on which the ligands are arranged connections to the ligand is selected Such that the maximum in order to bring about the desired ligand-orienting result, spatial distance between two adjacent ligands is no more and thus produce a multibinding compound. than 100 A. 0311 For example, different orientations can be achieved 0316. An example of a linker as presented by the grid is by including in the framework groups containing mono- or shown below for a biphenyl construct. polycyclic groups, including aryl and/or heteroaryl groups, or structures incorporating one or more carbon-carbon mul tiple bonds (alkenyl, alkenylene, alikyyl or alkynylene 8 7 ...... H H H groups). Other groups can also include oligomers and poly 6 ------mers which are branched- or straight-chain species. In 5 ------4 preferred embodiments, rigidity is imparted by the presence 3 of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, hetero 2. H 1 cyclic, etc.). In other preferred embodiments, the ring is a six O or ten member ring. In still further preferred embodiments, H H. H. C the ring is an aromatic ring Such as, for example, phenyl or naphthyl. 0312 Different hydrophobic/hydrophilic characteristics 0317 Nodes (1,2), (2,0), (4.4), (5.2), (4.0), (6.2), (7,4), of the linker as well as the presence or absence of charged (9.4), (10.2), (9,0), (7.0) all represent carbon atoms. Node moieties can readily be controlled by the skilled artisan. For (10.0) represents a chlorine atom. All other nodes (or dots) example, the hydrophobic nature of a linker derived from are points in space (i.e., represent an absence of atoms). hexamethylene diamine (HN(CH2)NH) or related 0318 Nodes (1.2) and (9.4) are attachment points. polyamines can be modified to be substantially more hydro Hydrogen atoms are affixed to nodes (2.4), (4.4), (4.0), (2.0), philic by replacing the alkylene group with a poly(oxyalky (7.4), (10.2) and (7.0). Nodes (5.2) and (6.2) are connected lene) group Such as found in the commercially available single bond. “Jeffamines. 0319. The carbon atoms present are connected by either 0313 Different frameworks can be designed to provide a single or double bonds, taking into consideration the preferred orientations of the ligands. Such frameworks may principle of resonance and/or tautomerism. be represented by using an array of dots (as shown below) wherein each dot may potentially be an atom, such as C, O. 0320 The intersection of the framework (linker) and the N. S. P. H. F. Cl, Br, and F or the dot may alternatively ligand group, and indeed, the framework (linker) itself can indicate the absence of an atom at that position. To facilitate have many different bonding patterns. Examples of accept the understanding of the framework structure, the frame able patterns of three contiguous atom arrangements are work is illustrated as a two dimensional array in the follow shown in the following diagram: US 2007/0134729 A1 Jun. 14, 2007 49

-continued O O O CCC NCC OCC SCC PCC | CCN NCN OCN SCN PCN CCO NCO OCO SCO PCO Ns-n1 N1 in N1 Ne-in-1 CCS NCS OCS SCS PCS O O CCP NCP OCP SCP PCP O CNC NNC ONC SNC PNC CNN NNN ONN SNN PNN S S S CNO NNO ONO SNO PNO N1 n1 N1 n1 N1 n1 CNS NNS ONS SNS PNS O CNP NNP ONP SNP PNP COC NOC OOC SOC POC O us S COO NON OON SON PON N1 n1 No N1 Ne-in-1 COC NOO OOO SOO POO O COP NOP OOS SOS POS OOP SOP POP S C CSC NSC N-1 n1 N1 n1 No1 no-1 CSN NSN OSC SSC PSC | CSO NSO OSN SSN PSN CSS NSS OSO SSO PSO O CSP NSP OSS SSS PSS OSP SSP PSP IS N J) N-N-1 CPC NPC N-sn 1 N CPN NPN OPC SPC PPC S C C CPO NPO OPN SPN PPN N1 no-1 N1 ns1 N1 no-1 CPS NPS OPO SPO PPO CPP NPP OPS SPS PPS OPP SPP PPP

0321) One skilled in the art would be able to identify bonding patterns that would produce multivalent com P P pounds. Methods for producing these bonding arrangements N N -Ns Ne-in-1O N1inO 1 are described in March, “Advanced Organic Chemistry, 4th N Edition, Wiley-Interscience, New York, N.Y. (1992). These O N arrangements are described in the grid of dots shown in the N )— scheme above. All of the possible arrangements for the five No-N-1 NN most preferred atoms are shown. Each atom has a variety of O acceptable oxidation states. The bonding arrangements underlined are less acceptable and are not preferred. The identification of an appropriate framework geometry 0322 Examples. of molecular structures in which the and size for ligand domain presentation are important steps above bonding patterns could be employed as components in the construction of a multibinding compound with of the linker are shown below. enhanced activity. Systematic spatial searching strategies can be used to aid in the identification of preferred frame works through an iterative process. FIG. 1 illustrates a usefull Strategy for determining an optimal framework dis play orientation for ligand domains. Various other strategies are known to those skilled in the art of molecular design and N-S-N-N-N-seO r r) can be used for preparing compounds of this invention. C 0323. As shown in FIG. 1 , display vectors around HN1 n1 similar central core structures such as a phenyl structure No1 C n1 N1 C n1 (Panel A) and a cyclohexane structure (Panel B) can be es varied, as can the spacing of the ligand domain from the core O structure (i.e., the length of the attaching moiety). It is to be No1 n1 N-N1 N1's 1 noted that core structures other than those shown here can be O O used for determining the optimal framework display orien tation of the ligands. The process may require the use of N1 n1 N N1 No N1 multiple copies of the same central core structure or com O O binations of different types of display cores. 0324. The above-described process can be extended to N-1N1ul N1 C & 1 N1ls n-1 trimers (FIG. 2) and compounds of higher valency (FIGS. 3 and 4). US 2007/0134729 A1 Jun. 14, 2007 50

0325 Assays of each of the individual compounds of a which allows their covalent attachment to the linker. Other collection generated as described above will lead to a subset lipophilic groups useful with the linkers of this invention of compounds with the desired enhanced activities (e.g., include fatty acid derivatives which do not form bilayers in potency, selectivity, etc.). The analysis of this Subset using aqueous medium until higher concentrations are reached. a technique such as Ensemble Molecular Dynamics will provide a framework orientation that favors the properties 0331. Also within the scope of this invention is the use of desired. A wide diversity of linkers is commercially avail ancillary groups which result in the multibinding compound able (see, e.g., Available-Chemical Directory (ACD)). Many being incorporated or anchored into a vesicle or other of the linkers that are suitable for use in this invention fall membranous structure such as a liposome or a micelle. The into this category. Other can be readily synthesized by term “lipid refers to any fatty acid derivative that is capable methods well known in the art and/or are described below. of forming a bilayer or a micelle such that a hydrophobic 0326 Having selected a preferred framework geometry, portion of the lipid material orients toward the bilayer while the physical properties of the linker can be optimized by a hydrophilic portion orients toward the aqueous phase. varying the chemical composition thereof. The composition Hydrophilic characteristics derive from the presence of of the linker can be varied in numerous ways to achieve the phosphate, carboxylic, Sulfate, amino, Sulfhydryl, nitro and desired physical properties for the multibinding compound. other like groups well known in the art. Hydrophobicity could be conferred by the inclusion of groups that include, 0327. It can therefore be seen that there is a plethora of but are not limited to, long chain Saturated and unsaturated possibilities for the composition of a linker. Examples of aliphatic hydrocarbon groups of up to 20 carbon atoms and linkers include aliphatic moieties, aromatic moieties, steroi Such groups Substituted by one or more aryl, heteroaryl, dal moieties, peptides, and the like. Specific examples are cycloalkyl, and/or heterocyclic group(s). Preferred lipids are peptides or polyamides, hydrocarbons, aromatic groups, phosphglycerides and sphingolipids, representative ethers, lipids, cationic or anionic groups, or a combination examples of which include phosphatidylcholine, phosphati thereof. dylethanolamine, phosphatidylserine, phosphatidylinositol, 0328 Examples are given below, but it should be under phosphatidic acid, palmitoyleoyl phosphatidylcholine, lyso stood that various changes may be made and equivalents phosphatidylcholine, lysophosphatidyl-ethanolamine, may be. Substituted without departing from the true spirit dipalmitoylphosphatidylcholine, dioleoylphosphatidylcho and scope of the invention. For example, properties of the lirne, distearoyl-phosphatidylcholine or dilinoleoylphos linker can be modified by the addition or insertion of phatidylcholine could be used. Other compounds lacking ancillary groups into or onto the linker, for example, to phosphorus, Such as sphingolipid and glycosphingolipid change the solubility of the multibinding compound (in families are also within the group designated as lipid. water, fats, lipids, biological fluids, etc.), hydrophobicity, Additionally, the amphipathic lipids described above may be hydrophilicity, linker flexibility, antigenicity, stability, and mixed with other lipids including triglycerides and sterols. the like. For example, the introduction of one or more 0332) The flexibility of the linker can be manipulated by poly(ethylene glycol) (PEG) groups onto or into the linker the inclusion of ancillary groups which are bulky and/or enhances the hydrophilicity and water solubility of the rigid. The presence of bulky or rigid groups can hinder free multibinding compound, increases both molecular weight rotation about bonds in the linker or bonds between the and molecular size and, depending on the nature of the linker and the ancillary group(s) or bonds between the linker unPEGylated linker, may increase the in vivo retention time. and the functional groups. Rigid groups can include, for Further PEG may decrease antigenicity and potentially example, those groups whose conformational lability is enhances the overall rigidity of the linker. restrained by the presence of rings and/or multiple bonds 0329 Ancillary groups which enhance the water solubil within the group, for example, aryl, heteroaryl, cycloalkyl, ity/hydrophilicity of the linker and, accordingly, the result cycloalkenyl, and heterocyclic groups. Other groups which ing multibinding compounds are useful in practicing this can impart rigidity include polypeptide groups such as oligo invention. Thus, it is within the scope of the present inven or polyproline chains. tion to use ancillary groups such as, for example, Small 0333 Rigidity can also be imparted electrostatically. repeating units of ethylene glycols, alcohols, polyols (e.g., Thus, if the ancillary groups are either positively or nega glycerin, glycerol propoxylate, saccharides, including tively charged, the similarly charged ancillary groups will mono-, oligosaccharides, etc.), carboxylates (e.g., Small force the presenter linker into a configuration affording the repeating units of glutamic acid, acrylic acid, etc.), armines maximum distance between each of the like charges. The (e.g., tetraethylenepentamine), and the like) to enhance the energetic cost of bringing the like-charged groups closer to water solubility and/or hydrophilicity of the multibinding each other will tend to hold the linker in a configuration that compounds of this invention. In preferred embodiments, the maintains the separation between the like-charged ancillary ancillary group used to improve water Solubility/hydrophi groups. Further ancillary groups bearing opposite charges licity will be a polyether. will tend to be attracted to their oppositely charged coun 0330. The incorporation of lipophilic ancillary groups terparts and potentially may enter into both inter- and within the structure of the linker to enhance the lipophilicity intramolecular ionic bonds. This non-covalent mechanism and/or hydrophobicity of the multibinding compounds will tend to hold the linker into a conformation which allows described herein is also within the scope of this invention. bonding between the oppositely charged groups. The addi Lipophilic groups useful with the linkers of this invention tion of ancillary groups which are charged, or alternatively, include, by way of example only, aryl and heteroaryl groups bear a latent charge when deprotected, following addition to which, as above, may be either unsubstituted or substituted the linker, include deprotectation of a carboxyl, hydroxyl, with other groups, but are at least Substituted with a group thiol or amino group by a change in pH, oxidation, reduction US 2007/0134729 A1 Jun. 14, 2007

or other mechanisms known to those skilled in the art which each occurrence, as can X. However, a trimer can also be a result in removal of the protecting group, is within the scope radial multibinding compound comprising three ligands of this invention. attached to a central core, and thus represented as (L)-X. where the linker X could include, for example, an aryl or 0334 Rigidity may also be imparted by internal hydrogen cycloalkyl group. Illustrations of trivalent and tetravalent bonding or by hydrophobic collapse. compounds of this invention are found in FIGS. 2 and 3 0335 Bulky groups can include, for example, large respectively where, again, the shaded circles represent atoms, ions (e.g., iodine, Sulfur, metal ions, etc.) or groups ligands. Tetravalent compounds can be represented in a containing large atoms, polycyclic groups, including aro linear array, e.g... matic groups, non-aromatic-groups and structures incorpo rating one or more carbon-carbon multiple bonds (i.e., alkenes and alkynes). Bulky groups can also include oligo in a branched array, e.g., mers and polymers which are branched- or straight-chain species. Species that are branched are expected to increase the rigidity of the structure more per unit molecular weight gain than are straight-chain species. 0336. In preferred embodiments, rigidity is imparted by the presence of cyclic groups (e.g., aryl, heteroaryl, cycloalkyl, heterocyclic, etc.). In other preferred embodi (a branched construct analogous to the isomers of butane ments, the linker comprises one or more six-membered —n-butyl, iso-butyl, sec-butyl, and t-butyl) or in a tetrahe rings. In still further preferred embodiments, the ring is an dral array, e.g., aryl group Such as, for example, phenyl or naphthyl. L. L. 0337. In view of the above, it is apparent that the appro N / priate selection of a linker group providing Suitable orien X. tation, restricted/unrestricted rotation, the desired degree of hydrophobicity/hydrophilicity, etc. is well within the skill of the art. Eliminating or reducing antigenicity of the multib inding compounds described herein is also within the scope where X and L are as defined herein. Alternatively, it could of this invention. In certain cases, the antigenicity of a be represented as an alkyl, aryl or cycloalkyl derivative as multibinding compound may be eliminated or reduced by above with four (4) ligands attached to the core linker. use of groups such as, for example, poly(ethylene glycol). 0342. The same considerations apply to higher multib 0338. As explained above, the multibinding compounds inding compounds of this invention containing 5-10 ligands described herein comprise 2-10 ligands attached to a linker as illustrated in FIG. 4 where, as before, the shaded circles that attaches the ligands in Such a manner that they are represent ligands. However, for multibinding agents presented to the enzyme for multivalent interactions with attached to a central linker Such as aryl or cycloalkyl, there ligand binding sites thereon/therein. The linker spatially is a self-evident constraint that there must be sufficient constrains these interactions to occur within dimensions attachment sites on the linker to accommodate the number of defined by the linker. This and other factors increases the ligands present; for example, a benzene ring could not biological activity of the multibinding compound as com directly accommodate more than 6 ligands, whereas a multi pared to the same number of ligands made available in ring linker (e.g., biphenyl) could accommodate a larger monobinding form. number of ligands. 0339. The compounds of this invention are preferably 0343 Certain of the above described compounds may represented by the empirical Formula (L)(X), where L. X, alternatively be represented as cyclic chains of the form: p and q are as defined above. This is intended to include the several ways in which the ligands can be linked together in order to achieve the objective of multivalency, and a more / LN detailed explanation is described below. X X 0340. As noted previously, the linker may be considered N 1-1 as a framework to which ligands are attached. Thus, it should be recognized that the ligands can be attached at any Suitable position on this framework, for example, at the and variants thereof termini of a linear chain or at any intermediate position. 0341 The simplest and most preferred multibinding 0344 All of the above variations are intended to be compound is a bivalent compound which can be represented within the scope of the invention defined by the Formula as L-X-L, where each L is independently a ligand which (L)(X), may be the same or different and each X is independently the 0345 Additionally, the linker moiety can be optionally linker. Examples of such bivalent compounds are provided Substituted at any atom therein by one or more alkyl, in FIG. 1 where each shaded circle represents a ligand. A Substituted alkyl, cycloalkyl, Substituted cycloalkyl, alkenyl, trivalent compound could also be represented in a linear Substituted alkenyl, cycloalkenyl, Substituted cycloalkenyl, fashion, i.e., as a sequence of repeated units L-X-L- alkynyl, Substituted alkynyl, aryl, heteroaryl and heterocy X-L, in which L is a ligand and is the same or different at clic group. US 2007/0134729 A1 Jun. 14, 2007 52

0346. In view of the above description of the linker, it is low solubility, hydrophobicity, hydrophilicity) may be ratio understood that the term “linker when used in combination nally modulated in multibinding forms, providing com with the term “multibinding compound” includes both a pounds with physical properties consistent with the desired covalently contiguous single linker (e.g., L X—L) and utility. multiple covalently non-contiguous linkers (L-X-L- Orientation: Selection of Ligand Attachment Points and X-L) within the multibinding compound. Linking Chemistry Combinatorial Libraries 0351. Several points are chosen on each ligand at which 0347 The methods described above lend themselves to to attach the ligand to the linker. The selected points on the combinatorial approaches for identifying multimeric com ligand/linker for attachment are functionalized to contain pounds which possess multibinding properties. complementary reactive functional groups. This permits probing the effects of presenting the ligands to their recep 0348 Specifically, factors such as the proper juxtaposi tor(s) in multiple relative orientations, an important multi tion of the individual ligands of a multibinding-compound binding design parameter. The only requirement for choos with respect to the relevant array of binding sites on a target ing attachment points is that attaching to at least one of these or targets is important in optimizing the interaction of the points does not abrogate activity of the ligand. Such points multibinding compound with its target(s) and to maximize for attachment can be identified by structural information the biological advantage through multivalency. One when available. For example, inspection of a co-crystal approach is to identify a library of candidate multibinding structure of a protease inhibitor bound to its target allows compounds with properties spanning the multibinding one to identify one or more sites where linker attachment parameters that are relevant for a particular target. These will not preclude the enzyme: inhibitor interaction. Alterna parameters include: (I) the identity of ligand(s), (2) the tively, evaluation of ligand/target binding by nuclear mag orientation of ligands, (3) the Valency of the construct, (4) netic resonance will permit the identification of sites non linker length, (5) linker geometry, (6) linker physical prop essential for ligand/target binding. See, for example, Fesik, erties, and (7) linker chemical functional groups. et al., U.S. Pat. No. 5,891,643. When such structural infor 0349 Libraries of multimeric compounds potentially mation is not available, utilization of structure-activity rela possessing multibinding properties (i.e., candidate multib tionships (SAR) for ligands will suggest positions where inding compounds) and comprising a multiplicity of Such Substantial structural variations are and are not allowed. In variables are prepared and these libraries are then evaluated the absence of both structural and. SAR information, a via conventional assays corresponding to the ligand selected library is merely selected with multiple points of attachment and the multibinding parameters desired. Considerations to allow presentation of the ligand in multiple distinct relevant to each of these variables are set forth below: orientations. Subsequent evaluation of this library will indi cate what positions are Suitable for attachment. Selection of ligand(s): 0352. It is important to emphasize that positions of 0350 A single ligand or set of ligands is (are) selected for attachment that do abrogate the activity of the monomeric incorporation into the libraries of candidate multibinding ligand may also be advantageously included in candidate compounds which library is directed against a particular multibinding compounds in the library provided that Such biological target or targets. The only requirement for the compounds bear at least one ligand attached in a manner ligands chosen is that they are capable of interacting with the which does not abrogate intrinsic activity. This selection selected target(s). Thus, ligands may be known drugs, modi derives from, for example, heterobivalent interactions fied forms of known drugs, Substructures of known drugs or within the context of a single target molecule. For example, substrates of modified forms of known drugs (which are consider a receptor antagonist ligand bound to its target competent to interact with the target), or other compounds. receptor, and then consider modifying this ligand by attach Ligands are preferably chosen based on known favorable ing to it a second copy of the same ligand with a linker which properties that may be projected to be carried over to or allows the second ligand to interact with the same receptor amplified in multibinding forms. Favorable properties molecule at sites proximal to the antagonist binding site, include demonstrated safety and efficacy in human patients, which include elements of the receptor that are not part of appropriate PK/ADME profiles, synthetic accessibility, and the formal antagonist binding site and/or elements of the desirable physical properties such as solubility, log P. etc. matrix Surrounding the receptor Such as the membrane. However, it is crucial to note that ligands which display an Here, the most favorable orientation for interaction of the unfavorable property from among the previous list may second ligand molecule with the receptor/matrix may be obtain a more favorable property through the process of achieved by attaching it to the linker at a position which multibinding compound formation; i.e., ligands should not abrogates activity of the ligand at the formal antagonist necessarily be excluded on Such a basis. For example, a binding site. Another way to consider this is that the SAR of ligand that is not sufficiently potent at a particular target so individual ligands within the context of a multibinding as to be efficacious in a human patient may become highly structure is often different from the SAR of those same potent and efficacious when presented in multibinding form. ligands in momomeric form. A ligand that is potent and efficacious but not of utility because of a non-mechanism-related toxic side effect may 0353. The foregoing discussion focused on bivalent inter have increased therapeutic index (increased potency relative actions of dimeric compounds bearing two copies of the to toxicity) as a multibinding compound. Compounds that same ligand joined to a single linker through different exhibit short in vivo half-lives may have-extended half-lives attachment points, one of which may abrogate the binding/ as multibinding compounds. Physical properties of ligands activity of the monomeric ligand. It should also be under that limit their usefulness (e.g. poor bioavailability due to stood that bivalent advantage may also be attained with US 2007/0134729 A1 Jun. 14, 2007

heterodimeric constructs bearing two different ligands that Linker Geometry and Rigidity: bind to common or different targets. For example, a 5HT receptor antagonist and a bladder-selective muscarinic M. 0358. The combination of ligand attachment site, linker antagonist may be joined to a linker through attachment length, linker geometry, and linker rigidity determine the points which do not abrogate the binding affinity of the possible ways in which the ligands of candidate multibind monomeric ligands for their respective receptor sites. The ing compounds may be displayed in three dimensions and dimeric compound may achieve enhanced affinity for both thereby presented to their binding sites. Linker geometry receptors due to favorable interactions between the 5HT and rigidity are nominally determined by chemical compo ligand and elements of the M receptor proximal to the formal Mantagonist binding site and between the M ligand sition and bonding pattern, which may be controlled and are and elements of the 5HT receptor proximal to the formal systematically varied as another spanning function in a 5HT, antagonist binding site. Thus, the dimeric compound multibinding array. For example, linker geometry is varied may be more potent and selective antagonist of overactive by attaching two ligands to the ortho, meta, and para bladder and a Superior therapy for urinary urge incontinence. positions of a benzene ring, or in cds- or trans-arrangements at the 1.1 - vs. 1.2- vs. 1.3- vs. 1.4- positions around a 0354) Once the ligand attachment points have been cho cyclohexane core or in cis- or trans-arrangements at a point sen, one identifies the types of chemical linkages that are possible at those points. The most preferred types of chemi of ethylene unsaturation. Linker rigidity is varied by con cal linkages are those that are compatible with the overall trolling the number and relative energies of different con structure of the ligand (or protected forms of the ligand) formational states possible for the linker. For example, a readily and generally formed, stable and intrinsically inocu divalent compound bearing two ligands joined by 1.8-octyl ous under typical chemical and physiological conditions, linker has many more degrees of freedom, and is therefore and compatible with a large number of available linkers. less rigid than a compound in which the two ligands are Amide bonds, ethers, amines, carbamates, ureas, and Sul attached to the 4.4 positions of a biphenyl linker. fonamides are but a few examples of preferred linkages. Linker Physical Properties: Linkers: Spanning Relevant Multibinding Parameters Through Selection of Valency, Linker Length, Linker Geom 0359 The physical properties of linkers are nominally etry, Rigidity, Physical Properties, and Chemical Functional determined by the chemical constitution and bonding pat Groups terns of the linker, and linker physical properties impact the 0355. In the library of linkers employed to generate the overall physical properties of the candidate multibinding library of candidate multibinding compounds, the selection compounds in which they are included. A range of linker of linkers employed in this library of linkers takes into compositions is typically selected to provide a range of consideration the following factors: physical properties (hydrophobicity, hydrophilicity, amphiphilicity, polarization, acidity, and basicity) in the Valency: candidate multibinding compounds. The particular choice of 0356. In most instances the library of linkers is initiated linker physical properties is made within the context of the with divalent linkers. The choice of ligands and proper physical properties of the ligands they join and preferably juxtaposition of two ligands relative to their binding sites the goal is to generate molecules with favorable PK/ADME permits such molecules to exhibit target binding affinities properties. For example, linkers can be selected to avoid and specificities more than Sufficient to confer biological those that are too hydrophilic or too hydrophobic to be advantage. Furthermore, divalent linkers or constructs are also typically of modest size such that they retain the readily absorbed and/or distributed in vivo. desirable biodistribution properties of small molecules. Linker Chemical Functional Groups: Linker Lengh 0360 Linker chemical functional groups are selected to 0357 Linkers are chosen in a range of lengths to allow be compatible with the chemistry chosen to connect linkers the spanning of a range of inter-ligand distances that encom to the ligands and to impart the range of physical properties pass the distance preferable for a given divalent interaction. Sufficient to span initial examination of this parameter. In some instances the preferred distance can be estimated rather precisely from high-resolution structural information Combinatorial Synthesis of targets, typically enzymes and soluble receptor targets. In other instances where high-resolution structural information 0361 Having chosen a set of n ligands (n being deter is not available (such as 7TM G-protein coupled receptors), mined by the sum of the number of different attachment one can make use of simple models to estimate, the maxi points for each ligand chosen) and m linkers by the process mum distance between binding sites either on adjacent outlined above, a library of (n!)m candidate divalent multi receptors or at different locations on the same receptor. In binding compounds is prepared which spans the relevant situations where two binding sites are present on the same target (or target Subunit for multisubunit targets), preferred multibinding design parameters for a particular target. For linker distances are 2-20 A, with more preferred linker example, an array generated from two ligands, one which distances of 3-12 A. In situations where two binding sites has two attachment points (A1, A2) and one which has three reside on separate (e.g., protein) target sites, preferred linker attachment points (B 1, B2, B3) joined in all possible distances are 20-100 A, with more preferred distances of combinations provide for at least 15 possible combinations 30-70 A. of multibinding compounds: US 2007/0134729 A1 Jun. 14, 2007 54

0362. When each of these combinations is joined by 10 untagged libaries of candidate multivalent compounds by different linkers, a library of 150 candidate multibinding methods known in the art such as those described by compounds results. Hindsgaul, et al., Canadian Patent Application No. 2,240, 325 which was published on Jul. 11, 1998. Such methods 0363 Given the combinatorial nature of the library, com couple frontal affinity chromatography with mass spectros mon chemistries are preferably used to join the reactive copy to determine both the structure and relative binding functionalities on the ligands with complementary reactive affinities of candidate multibinding compounds to receptors. functionalities on the linkers. The library therefore lends itself to efficient parallel synthetic methods. The combina 0367 The process set forth above for dimeric candidate torial library can employ solid phase chemistries well known multibinding compounds can, of course, be extended to in the art wherein the ligand and/or linker is attached to a trimeric candidate compounds and higher analogs thereof. solid support. Alternatively and preferably, the combinato rial libary is prepared in the solution phase. After synthesis, Follow-up Synthesis and Analysis of Additional Array(s): candidate multibinding compounds are optionally purified 0368 Based on the information obtained through analy before assaying for activity by, for example, chromato sis of the initial library, an optional component of the graphic methods (e.g., HPLC). process is to ascertain one or more promising multibinding “lead compounds as defined by particular relative ligand Analysis of Array by Biochemical, Analytical, Pharmaco orientations, linker lengths, linker geometries, etc. Addi logical, and Computational Methods: tional libraries can then be generated around these leads to 0364 Various methods are used to characterize the prop provide for further information regarding structure to activ erties and activities of the candidate multibinding com ity relationships. These arrays typically bear more focused pounds in the library to determine which compounds possess variations in linker structure in an effort to further optimize multibinding properties. Physical constants such as solubil target affinity and/or activity at the target (antagonism, ity under various solvent conditions and logD/clogD values partial agonism, etc.), and/or alter physical properties. By are determined. A combination of NMR spectroscopy and iterative redesign/analysis using the novel principles of computational methods is used to determine low-energy multibinding design along with classical medicinal chemis conformations of the candidate multibinding compounds in try, biochemistry, and pharmacology approaches, one is able fluid media. The ability of the members of the library to bind to prepare and identify optimal multibinding compounds to the desired target and other targets is determined by that exhibit biological advantage towards their targets and as various standard methods, which include radioligand dis therapeutic agents. placement assays for receptor and ion channel targets, and 0369 To further elaborate upon this procedure, suitable kinetic inhibition analysis for many enzyme targets. In vitro divalent linkers include, by way of example only, those efficacy, such as for receptor agonists and antagonists, ion derived from dicarboxylic acids, disulfonylhalides, dialde channel blockers, and antimicrobial activity, are determined. hydes, diketones, dihalides, diisocyanates.diamines, diols, Pharmacological data, including oral absorption, everted gut mixtures of carboxylic acids, Sulfonylhalides, aldehydes, penetration, other pharmacokinetic parameters and efficacy ketones, halides, isocyanates, amines and diols. In each case, data are determined in appropriate models. In this way, key the carboxylic acid, sulfonylhalide, aldehyde, ketone, halide, structure-activity relationships are obtained for multibinding isocyanate, amine and diol functional group is reacted with design parameters which are then used to direct future work. a complementary functionality on the ligand to form a 0365. The members of the library which exhibit multib covalent linkage. Such complementary functionality is well inding properties, as defined herein, can be readily deter known in the art as illustrated in the following table: mined by conventional methods. First those members which exhibit multibinding properties are identified by conven Complementary Binding Chemistries tional methods as described above including conventional 0370 assays (both in vitro and in vivo). 0366 Second, ascertaining the structure of those com pounds which exhibit multibinding properties can be accom plished via art recognized procedures. For example, each First Reactive Group Second Reactive Group Linkage member of the library can be encrypted or tagged with carboxyl amine amide Sulfonyl halide amine Sulfonamide appropriate information allowing determination of the struc hydroxyl alkyl/aryl halide ether ture of relevant members at a later time. See, for example, hydroxyl isocyanate urethane Dower, et al., International Patent Application Publication amine epoxide 3-hydroxyamine amine alkyl/aryl halide alkylamine No. WO 93/06121; Brenner, et al., Proc. Natl. Acad. Sci., hydroxyl carboxyl ester USA, 89:5181 (1992); Gallop, et al., U.S. Pat. No. 5,846, amine aldehyde/NaCNBH amine 839; each of which are incorporated herein by reference in hydroxylamine Sulfonyl halide Sulfonamide its entirety. Alternatively, the structure of relevant multiva aldehyde amine/NaCHBH amine lent compounds can also be determined from soluble and US 2007/0134729 A1 Jun. 14, 2007 55

X-418) that can be used to prepare linkers incorporated in -continued the multibinding compounds of this invention utilizing the First Reactive Group Second Reactive Group Linkage chemistry described above. For example, 1,10-decanedicar aldehyde amine/NaCHBH amine boxylic acid, X1, can be reacted with 2 equivalents of a amine isocynate le:8 ligand carrying an amino group in the presence of a coupling reagent Such as DCC to provide a multibinding compound of 0371) The following table illustrates, by way of formula (I) wherein the ligands are linked via 1,10-de examples, starting materials (identified as X-1 through canediamido linking group.

Diacids X-1 X-2 OH

HO C H3

HO cO X-3 X-4 O CHOH

O S - ( O y U/ S / OH O HO H3C

CH X-5 X-6 O OH

HO O CH3

OH

X-9 X-10 OH OH OH OH N < x^. HO CH

N HO HO O X-11 X-12 HO O OH

HO CH3 X-14 OH

US 2007/0134729 A1 Jun. 14, 2007

-continued

X-28 X-29 O O O

HO OH HO OH O O O

X-30 X-31 HO O O

O n HO--- S Ns O CH OH OH

X-32 X-33 CHChiral Chiral

O O O O

O O ---,s OH HO---,s OH

O O

X-34 X-35 Chiral O O O OH HO HO OH O C HC CH3 X-36 X-37 O OH O

HO ),F O HO O F OH X-38 X-39 HO O OH

M CH3 US 2007/0134729 A1 Jun. 14, 2007

-continued

X-40 X-41 OH O M CH3 Chiral O O OH O O O O OH OH O C H3 V N OS 1

HC1 N NCH,

X-42 X-43 O O OH OH --~~~~s. O

OH

X-44 X-45 HO HO O CH3 O O HO

H3C O OH O

X-46 X-47 Chiral OH O u O HO 8. CH3 HO S) O S O

H3C OH X-48 X-49 O OH O CH O

OH US 2007/0134729 A1 Jun. 14, 2007

-continued X-50 X-51 O OH

HO O

O X-52 X-53 Chiral HN Chiral OH N 2s, O O O 1.

O OH HO O HO X-54 X-55 Chiral O "V HO --- Snu1a-1\ S O O . OH OH HO X-56 X-57 O O OH OH O O OH OH CH, O O 1. O HC O OH

HC

X-58 X-59 V Chiral \ y OH HO . O O

X-61 X-60 O OH HO HO--~- r N--- O S HO

X-62 X-63

Chiral H3C CH3 O OH

HO S S O

US 2007/0134729 A1 Jun. 14, 2007 61

-continued

X-78 O OH O

OH

X-79 O

HO CH

OH

O

X-8O X-81 Chiral O Chiral H3 C N " ul O N CH3 O OH r C H3 O

OH OH

X-82 Chiral O OH

HO HO OH

O HO N-N-N Sr- OH O O X-84 O

OH

HO O X-85 O

O HO

OH US 2007/0134729 A1 Jun. 14, 2007

-continued X-86 X-87 CH Chiral H

O CH3 O HO 2. No, OH & 4, a. OH O H

HO O X-88 "Sr. X-89 O C NNus. O X-90 X-91 O O Ps lu O O n-rr HO O

O

X-92 X-93 O Chiral O OH

OH HO r O S

CH3 OH X-94 X-95 O O

HO OH

X-96 X-97 9 F F F F F F F F F F OH OH

OH US 2007/0134729 A1 Jun. 14, 2007

-continued X-98 X-99 H3C CH3 H3C CH3 HO OH

O O

X-100 X-101

X-102 X-103 F F F F F F F F HO OH

F F F F F F )

X-104 X-105 O OH -- S N1 NullsO

X-106 X-107 O

N OH

OH OH N OH O O

C X-108 X-109 O OH

OH HO

OH O

C US 2007/0134729 A1 Jun. 14, 2007

-continued X-110 X-111 Chiral O O O N OH HO 2. OH C OH N

O O

Br X-112 X-113 O Chiral O

HO OH OH O

X-114 X-115 Chiral

OH

OH

O

X-116 X-117 Chiral O

HO N O O Y O - N - - OH Ó

O OH O

O X-118 X-119 OH HO O N OH O so e ... / OH O X-12O X-121 O OH O OH US 2007/0134729 A1 Jun. 14, 2007

-continued

X-122 X-123 O O

HO N N-/ OH

O O

X-124 X-125 O O O O O HO OH OH NX=o OH HN

X-126 X-127 OH HO CHChiral

O O O

O N OH N

O O

HO O

X-128 X-129 O OH OH HO--~~s. HO 1N1 NullsO O X-130 X-131 O H

OH O H. CH ''', OH HO O H HO O X-132 O OH O

OH

OH

US 2007/0134729 A1 Jun. 14, 2007 68

-continued X-160 X-161 ro X-162 X-163

X-164 X-165

X-166 X-167

X-168 X-169

X-170 X-171

X-172 X-173

US 2007/0134729 A1 Jun. 14, 2007 70

-continued X-193 X-194 Br Br

X-195 X-196 CH

X-197 X-198 HO OH Br Br

X-199 Br

Br CHS 5 X-204 C

Cl X-205 Nu-1N1\-1N1 Br N-1-1N1 Br X-208 HC Br H3CNo Br HN Br Br X-209 X-210 "N-1s-1N C C

X-211 X-212 N-1N1 S-1-1N1 Br N-- Br CH3 X-213 X-214 C