CA2214789A1

CA2214789A1 - Carbopeptoids and carbonucleotoids

Info

Publication number: CA2214789A1
Application number: CA 2214789
Authority: CA
Inventors: Kyriacos C. Nicolaou
Original assignee: Individual
Current assignee: Scripps Research Institute
Priority date: 1995-03-08
Filing date: 1996-03-08
Publication date: 1996-09-12
Also published as: EP0827406A4; AU5188296A; AU717099B2; EP0827406A1; WO1996027379A1

Abstract

Librairies are synthesized with oligomeric carbopeptoids and carbonucleotoids.
Carbopeptoids are oligosaccharides having carbohydrate subunits linked to one another by amide bonds. Carbonucleotoids are oligosaccharides having carbohydrate subunits linked to one another by phosphodiester bonds.
Carbopeptoid librairies may be fabricated using automated polypeptide synthesizers. Carbonucleotoid librairies may be fabricated using automated polynucleotide synthesizers.

Description

W 096/27379 PCTn~S9~J~3227 r2~RRo~~ oIDs AND ~'ARRr~cLl iO ~ S

Specification Field of thç Invention:
The invention relates to oligosaccharides and libraries incorporating oligosaccharide. More particularly, the invention relates to oligosaccharides and libraries of oligosaccharides which employ amide and/or phosphodiester linkages for joining adjacent carbohydrate subunits.

Backaround:
Carbohydrates are known to mediate many cellular recognition processes. Carbohydrates can serve directly as binding molecules and, in such instances, are essential to the recognition process. A review of the biological role of carbohydrates with respect to cellular recognition phenomena is provided by Sharon et al (Scienti~ic American, January 1993, 82). The emerging importance of glycobiology is further characterized by Mekelburger et al. (Angew. Chem. Int. Ed. Engl. 1992, 31, 1571) and by Dagani et al. (Chem. Eng. News, February 1, 1993, 28).
Dysfunctional mediation of cellular recognition processes can lead to disease states. If a cellular recognition process is mediated by an oligosaccharide, then an absence or excess of such oligosaccharide can lead to a dvsfunctional mediation of such process. The SUBSTITUTE SHEET(RULE 26) W O 96/27379 PC~rrUS96/03227 mediating oligosaccharide may be deficient or absent due to a deficiency of production or due to a high rate of catabolism. If rate of catabolism is excessive, then catabolically resistant analogs of the bioactive oligosaccharide may be preferred as drug candidates as compared to the native bioactive oligosaccharide.
Accordingly, what is needed is a library which includes analogs of known bioactive oligosaccharides.
Such a library may be usefully employed for screening drug candidates.
Central requirements ~or the design of libraries of oligosaccharide analogs include the following:
(a) A need to maximize the potential of the designed oligosaccharides as ligand and drug candidates;
(b) A need to capitalize on existing highly sophisticated technology directed to the synthesis of oligopeptides and oligonucleotides in order to facilitate the rapid and efficient design and construction of oligosaccharides; and (c) A need for ~lexibility with respect to synthesizing either single target molecules or large libraries of target molecules simultaneously.
Methodologies for synthesizing biopolymers are well developed for peptides, nucleic acids, and saccharides.
Segments of oligopeptides and of oligonucleotides can now be routinely synthesized both in solution and in the solid phase, manually and/or on automated systems. The synthesis of such structures is ~acilitated by the SUBSTITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
WO 96127379 PCrlUS96103227 availability of ef~icient techniques and sophisticated instrumentation for synthesizing peptide and phosphate bonds with high yields. The synthesis of oligope~tides and oligonucleotides is also facilitated by the absence of stereocenters in these linkages. In contrast, technology for the construction of oligosaccharides is comparatively less sophisticated and efficient.
Synthetic methods ~or constructing oligosaccharides give comparatively lower yields and are complicated by the two isomer possibilities (~ and $) in glycoside bond formation.
Techniques and chemical methods ~or simultaneously synthesizing multiple oligopeptides, e.g. 100-150 completely differe~t peptides having lengths of up to 20 amino acid residues, are reviewed by Jung, G. et al.
(Angew. Chem, Int . Ed . ~ngl . 1992, 31, 367-383 -incorporated therein by reference). Such techniques facilitate the construction of oligopeptide libraries.
Simon, et al (Proc, Natl. Acad. Sci. USA, 1992, 89, 9367-9371) disclose oligopeptide analogs in which amino acid side chain groups are attached not to conventional peptide backbone carbons but to peptide backbone nitrogens. Such analogs are termed peptoids. Simon also discloses the construction of peptoid libraries as a modular approach to drug discovery. Simon's oligopeptoids are shown by calculation to have greater conformational freedom as compared to conventional oligopeptides. Accordingly, oligopeptoids are thought to have greater potential as p~rm~ceutically useful binding ~1-SUBSTITUTE SHEET (RULE 26) W 096/27379 PCTrUS96/03227 ligands as compared to conventional oligopeptides having close sequence homology to such oligopeptoids.
Von Roedern et al. disclose a carbohydrate amino acid (Angew. Chem, Int . Ed . Engl . 1994, 31, 687-689).
Although von Roedern discloses that carbohydrate amino acids may be coupled to peptides, he does not disclose that they may also be polymerized so as to form oligosaccharides.

Summarv:
A first aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbopeptoids (CPD's).
Glycopeptoids are preferred carbopeptoids. Carbopeptoids and glcopeptoids are oligosaccharides which employ peptide-like amide bonds for linking the various carbohydrate subunits within an oligomer assembly. Amide bond formation may be achieved by employing oligopeptide synthesis technology and instrumentation. The method allows for the design and synthesis of specific compounds for biological and pharmacological investigations. The method also allows for the generation of libraries of compounds for biological and ph~rm~sological screening.
Conventional screening techniques employed with respect to peptide and peptoid libraries (Simon et al., supra) may also be employed with respect to carbopeptoid libraries. The design takes advantage of the multifunctionality of carbohydrate subunits to maximize SUBSTITUTE SHEET (RULE 26) , ~ CA 02214789 1997-09-0~
W~96/27379 PCT~US96J03~7 the binding properties of the molecules. The ease and high efficiency by which the peptide-like linkages can be constructed make the synthesis of these molecules a practical proposition. Furthermore, non-carbohydrate units may be inserted into the sequence making this approach even more flexible and versatile for the generation o~ new libraries o~ organic compounds.
More particularly, the invention is directed to a oligomeric carbopeptoid or glycopeptoid compound having carbohydrate amino acid subunits (CA's) or glycoside amino acid subunits (GA's) coupled to one another via an amide linkage. The amide linkage may be represented by the formula CAl-(CO-NH)-CA2. The amide linkage (CO-NH) includes a carbonyl carbon and an amido nitrogen. A
first carbohydrate amino acid subunit CAl or glycoside amino acid subunit GAl has an anomeric carbon bonded to the carbonyl carbon of the amide linkage. The anomeric carbon of the ~irst carbohydrate amino acid subunit CAl forms a C-glycosidic bond with the carbonyl carbon of the amide linkage and maintains the carbohydrate in a closed ring configuration. A second carbohydrate amino acid subunit C~2 has a non-anomeric carbon bonded to the amido nitrogen of the amide linkage. The second carbohydrate amino acid subunit CA2, like the ~irst amino acid subunit CAl, may include an anomeric carbon bonded to the carbonyl carbon o~ a second amide linkage linking the second carbohydrate amino acid subunit CA2 to a third carbohydrate amino acid subunit CA3, etc. In this instance, the anomeric carbon of the second SUBSTITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
W 096/27379 PCTrU5~G~'~3227 carbohydrate amino acid subunit CA2 forms a C-glycosidic bond with the carbonyl carbon of the amide linkage and maintains the carbohydrate in a closed ring configuration. On the other hand, if the second carbohydrate amino acid subunit C~2 is a t~rm; n~ ~
subunit, then its anomeric carbon may form a hemiacetal, a hemiketal, or a glycoside.
The invention is also directed to a process for synthesizing the above oligomeric carbopeptoid or glycopeptoid compound. The synthetic process involves the coupling of two or more carbohydrate amino acid subunits (Q's) or glycoside amino acid subunits (GA's) to one another by means of amide linkages.
The invention is also directed to libraries of oligomeric carbopeptoid or glycopeptoid compounds. Such libraries are employable for drug screening. Each oligomeric carbopeptoid or glydopeptoid compound includes at least two carbohydrate amino acid subunits (CA's) or glycoside amino acid subunits (GA's) coupled to one another via an amide linkage as indicated above. The invention is also directed to an improved process ~or synthesizing the above library of oligomers. The process employs an elongation step for coupling the subunits to one another to produce the oligomers. In the elongation step, two carbohydrate amino acid subunits (CA's) or glycoside amino acid subunits (GA's) are coupled to one another via an amide linkage as indicated above.
The invention is also directed to chemical intermediates for producing oligomeric carbopeptoids. A

SUBSTITUTE SHEET (RULE 2B) , , ~ CA 02214789 1997-09-0~
W 096127379 PCTrUS96/D3227 first chemical intermediate is a derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons. The anomeric carbon is substituted with a carboxyl radical. Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl, blocked amino, differentially protected amino, and hydrogen, with the proviso that at least one radical is a differentially protected amino. A second chemical int~rm~iate is a derived carbohydrate amino acid similar to the first except that the non-anomeric carbons are substituted with a radical selected ~rom the group consisting of blocked hydroxyl, blocked amino, unprotected amino, and hydrogen, with the proviso that at least one radical is an unprotected amino and at least one radical is a blocked hydroxyl or amino.
A second aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbonucleotoids (CND's).
Carbonucleotoids are oligosaccharides which employ oligonucleotide-like phosphate bonds for linking the various carbohydrate subunits within an oligomer assembly. Phosphate bond formation may be achieved by employing technology and instrumentation developed for oligonucleotide synthesis. The phosphate bonds employed within carbonucleotoids are convenient linkages for coupling these units. The ease and high efficiency by which the oligonucleotide-like linkages can be constructed make the synthesis of these molecules a SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~ ' W 096/27379 PCT~US96/03227 practical proposition.
The disclosed methods are characterized by their versatility and practicality. The methods may exploit conventional solid phase and automated synthesis techniques for producing carbopeptoids and carbonucleotoids in large scale.
More particularly, the second aspect of the invention is directed to an oligomeric carbonucleotoid molecule comprising carbohydrate C-glycoside subunits (CG's) coupled to one another via a phosphodiester linkage. The phosphodiester linkage may be represented by the structure: CG1-C1 -(O-PO(OH)-O)-CG2. The first carbohydrate C-glycoside subunit (CG1-C1') has an anomeric carbon forming a C-glycosidic bond wilh a carbon C1'. In turn the carbon C1' is bonded to the phosphodiester linkage. The second carbohydrate C-glycoside subunit CG2 has a non-anomeric carbon bonded to the phosphodiester linkage. The invention is also directed a process for synthesizing the oligomeric carbonucleotoid molecule.
The process employs a coupling step wherein two or more carbohydrate C-glycoside subunits (CG's) are coupled by means of a phosphodiester linkage as indicated above.
The second aspect of the invention is also directed to libraries of oligomeric carbonucleotoid molecules.
The libraries are employable for drug screening. Each oligomeric carbonucleotoid molecule including at least two carbohydrate C-glycoside subunits (CG's) coupled to one another by means of a phosphodiester linkage as indicated above. The invention is also directed to an SUBSTITUTE SH EET (RULE 26) . ' CA 02214789 1997-09-0~
W 096/27379 PCTnUS9~n3227 improved process for synthesizing a library of oligomers.
The process employs an elongation step wherein subunits are coupled to one another to produce the oligomers. The improvement is directed to the use of phosphodiester linkage linkages for linking the C-glycoside subunits as indicated above.
The second aspect of the invention is also directed to derived carbohydrate C-glycosides having an anomeric carbon and non-anomeric carbons. ~he ~n~iC carbon forms a C-glycosidic bond with carbon C1 . In turn, the carbon Cîl is bonded to an phosphoramidite. Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl, differentizlly protected hydroxyl, and hydrogen, with the proviso that at least one radical is a differentially protected hydroxyl. An alternative derived carbohydrate C-glycoside is similar to the above except that each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl, unprotected hydrox~rl, and hydrogen, with the proviso that at least one radical is an unprotected hydroxyl and at least one radical is a blocked hydroxyl.

SUBSTITUTE SHEET (RULE 26) W 096/27379 PCTrUS96/03227 H ~~, H ~~, H ~~ ' H ~~' H
... , NO-~ , N o_~ , N o_~ , N _o_ , N--I carbopeptoid (CPD) H2N ~COOH H2N O_ COOH H2N O_ COOH H2N O COOH

~0~ ~0~ ~0 1 1 l l 1 1 l l 1 1 ~ o ~', P-' ~~0_/--~'' 'P-' ~~0_~--~''''P-' ~~o_~--~' ~f '0' ~ ~0' ~J ~0~ ~ ~
H _ H H
I l carbonucleotoid (CND) HO OH HO o_ OH HO O_ OH HO o ~OH

Scheme 1. Designed carbopeptoids (CPD's) and carbonucleotoids (CND's) SUBSTITUTE SHEET (RULE 26) , , ~ CA 02214789 1997-09-0~
W~96/27379 PCTnUS96/03~27 Detailed ~escri~tion:
Retrosynthetic sch~s for carbopeptoids (compound I) and carbonucleotoids (compound II) are illustrated in Scheme 1.
The carbopeptoids (CPD's) are oligomers having repeating carbohydrate subunits linked to one another by means of amide linkage units. More particularly, the carbonyl carbon of each amide linkage unit is bonded to the anomeric carbon of a carbohydrate subunit.
Similarly, the amide nitrogen of the amide linkage unit is bonded to a no~-anomeric carbon. The retrosynthetic scheme suggests that the amide bond may be split and that the preferred starting materials are carbohydrate amino acids.
Carbonucleotoids (CND's) are oligosaccharides in which carbohydrate C-glycoside subunits (CG's) are linked to one another by means of phosphodies~er bonds. More particularly, the retrosynthetic scheme suggests that the phosphate group may be eliminated, yielding hydroxylated starting material.
Scheme 2 illustrates representative carbohydrate amino acid subunits (CA's) and carbohydrate C-glycoside subunits (CG's). Preferred carbohydrate amino acid subunits (CA's) include the ~ollowing:
D-glucose having an unprotected carboxyl at the anomeric C(1) position, an unprotec~ed amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having an unprotected carboxyl at the SUBSTITUTE SHEET (RULE 26) .

CA 02214789 1997-09-05 ' ' anomeric C(l) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-galactose having an unprotected carboxyl at the anomeric C(l) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;

SUBSTITUTE SH EET (RULE 26) CA 022l4789 l997-09-05 W O 96J27379 PC~rrJS961~3227 sH2N ~ ~ COOH ~ O ~ COOH ~ ~ r COOH ~ O ~ COOH

TESO ~ 'OTES TESO ~ ~OTESTESO ~ "OTES TESO ~ NHAc OTES OTES OTES OTES
D-glucose (a + 0 D-mannose (a + ~) D-galactose (a + ~) N-acetyl-D-glucosa 2 4 6 mine (a + p) H2N ~ ~ ~COOH H2N ~ ~COOH~ ~'COOH HOOC r ~ ~ O-NH2 TESO ~ OTES TESO . OTES TES "OTES TESO ~ OTES
OTES OTES OTES OTES
a - D- idose a - D- altrosea - D- gulose D-glucose (a + ~) HOOC~O~O-NH2 HOOC~O~O-NH2 HOOC~O~O-NH2 TESO' ~ OTES TESO ~ 'OTES TESO' ~ 'NHAc OTES OTES OTES
D- mannose (~ + 0 D- galactose (o~ + o N-acetyl- D-gl~ c~minp(c~+~) 18 20 22 H2N ~ COOH H2N ~ COOH

TESO OlES TESO OTES
D- ribose D- arabinose ~ ~ O NPr2 ~ ~ ~ NPr2 TESO' ~ 'OTES TESO ~ "OTES
OTES OTES
D-glucose (c~ + ~) D-galactose (a + ~) HO ~ ~ O~ ~NiPr2 ~ ~ ~ NPr2 TESO ~ OTES TESO' ~ NHAc OTES OTES
D-mannose (a + ,B) N-acetyl-D-glll- nc~min,~ + ~) Scheme 2. StrLlctu}es of carbohydrate amirloacids (CA's) and C-glycosides (CG's) SUBSTITlJTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096t27379 PCTrUS96/03227 N-acetyl-D-glucosamine having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, a blocked amino group at the C(2) position, and blocked hydroxyls at the C(3) and C(4) positions;
~-D-idose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
~-D-altrose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
~-D-gulose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
~-D-glucose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having an unprotected O-glycosidic amino at the ~n~m~ric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positionsi D-galactose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;

SUBSTITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
W096127379 PCT~S96103227 N-acetyl-D-glucosamine having an unprotected O-glycosidic amino at the ~nom~ric C(1) position, an unprotected carboxyl as the C(6) position, a blocked amino group at the C(2) position and blocked hydroxyls at the C(3) and C(4) positions;
D-ribose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions; and D-arabinose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions.

Preferred carbohydrate amino acid subunits (CA's) include the ~ollowing:
D-glucose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-galactose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions; and N-acetyl-D-glucosamine having a C(1) C1 -glycosidic SUBSTITUTE SHEET (RULE 26) W 096/27379 PCTrUS96/03227 carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position, a blocked amino at the C(2) position, and blocked hydroxyls at the C(3) and C(4) positions.

Scheme 3 outlines a preferred synthesis of suitably protected carbohydrate amino acid subunits (CA's) from D-glucose, i.e. compound 46.

SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096/27379 PCTnUS~GM3227 AcO~ ~ 1. TMSCN, SnCll ~OrCOOEt AcO' 'OAc 2. EtOH, H2S04 (cat) HO" y 'OH
OAc OH 38 1. PivC1, DMAP, pyridine y 2. Et3SiOTf, iPr2EtN, CH2C12 N3~ r 1 NaOEt, EtOH PivO--~OrCOOEt TESO" ~ "OTES 2. TDHPFA, DIAD, Ph3P TESO' ~ 'OTES
OTES (41~2) OTES

1. H~, Pd on C
~I

H2N ~ COOEt FMOCN ~ COOH
~~ ~ 1. NaOH, p-dioxane l r TESO' y 'OTES 2. FMOC-Cl, NaHCO3, H~O TESO ~ 'OTES
OTES OTES

Scheme 3. Svnthesis of glucose-derived carbohydrate amino acid (CA): 46 SUBSTITUTE SHEET (RULE 26) W096/27379 PCTrUS96103227 Scheme 4 outlines the synthesis of suitably protected carbohydrate amino acid subunits (CA's) ~rom N-acetyl-D-glucosamine, i.e. compound 62.

SUBSTITUTE SH EET (RULE 26) WO 96/27379 PCTtUS96~03227 AcO--~ r L TMSCN, SnCl4 ~OrCOOEt AcO' ~ 'NPhth 2. EtOH, H2SO~L (cat) HO' ~ 'NHAc 3. Ac2O, MeOH
OAc OH 54 1. PivCl, DMAP, pyridirle ~, 2. Et3SiOTf, 'Pr2EtN, CH2Ck N3~ r 1 NaOEt EtOH PivO~OrCOOEt TESO" I "NHAc 2. DPPA DIAD, Ph3P TESO" 'NHAc OTES (57-58) OTES

l.H2,PdonC

H2N ~~ COOEt Fl\/lOCN~O~ COOH
~ 1. NaOH p-dioxane ¦ ~
TESO' ~ "NHAc 2. FMOC-Cl, NaHCO3, H20 ~ 'NHAc OTES (61-62) OTES

Scheme 4. Synthesis of N-acetyl~ s~mine-derived amino acid (CA): 62 SUBSTITUTE SHEET (RULE 26) W096/27379 CA 02214789 1gs7-os-05 PCT~S96/03227 Scheme 5 summarizes the synthesis of hexamer 74, i.e glucose-glucosamine hetero carbopeptoid (CPD).

SIJBSTITUTE SHEET (RIJLE 26) CA 022l4789 l997-09-05 W096J27379 P~l~U~,G,'03227 o ~ z~ $

O =~ O t~ O Z~ O U . O =~ O

C/~ ~~ ~
O ~ ~ Z C

a ~
~ I 0=~ ~~

~ 0~ 0 ,_ ~ \ /

-- LL ~ G

X
-. _ ..

C~
U) SUBSTITUTE SHEET (RULE 26) W 096/27379 PCT~US96103227 0~0 ~~~ ~~~ ~~

~z Z r' =~Z G =~ ~=~

T ~ 0~0 ~ ~~ ~- =

~0 ~~~'~r ~ O,, ~ ~~ O = ~ o 0~ l o=~ O
~0 o~o 0~0 ~~0 T =~--0 ~--0 o~ T

0=~ 0 ~=~ ~

o \>_ O O '>--O
~/ ~ ~/ I

I I

5, _G
~ SUBST~TUTE Sl IEET (RULE 26) W096/27379 PCTn~S9~0322 Scheme 6 illustrates the constructio~ of suitably protected and activated C-glycoside subunits (CG's~
corresponding to glucose.

-SUBSTITl3TE SHEET (RULE 26) W 096/27379 PCTrU5~5J~227 ACO ~ ~ ~ scN~sncl4 HO ~ O r COOEt AcO~ ''OAc 2.EtOH,H2SO4(cat) HO~ 'OH
OAc OH 76 1.~im~th~xytritylrhl~n~, DMAP, DMF
~ 2. Et3SiOTf, iP}2EtN, CH2C12 DMTO ~ O ~ OH iBU2A1H,CH2Cl2 DMTO ~ ~ r COOEt TESO' ~ 'OTES TESO' 'OTES
OTES OTES
1. Et3SiCl, imid., DMF 78 OCH2CH2CN ~Cl3CCOOH, CH3NO2, THF

Cl N Pr2 \~
Pr2EtN, CH2Cl2 OCH2CH2CN

DMTO ~ ~ ~ ~P~Nipr2 DMTO ~ O ~ OTES

TESO~ y 'OTES TESO~' y 'OTES
OTES OTES

Scheme 6. Synthesis of glucose-derived C-glycoside (CG) SUBSTITUTE SHEET (RULE 26) W096/2~379 PCTrUS96103227 Scheme 7 illustrates the construction of suitably protected and activated C-glycoside subunits (CG's) corresponding to glucosamine.
-SUBSTITUTE SHEET (RULE 26) W096/27379 P~l/u~ 03227 AcO ~ ~ scN~sncl4 HO ~ O r COOEt AcO' ~ 3 Ac20, MeOH HO' 'NHAC
OAc OH 86 m~thnxytrityl chloride, DMAP, DMF
~ 2. Et3SiOTf, iPr2EtN, CH2Cl2 DMTO~ ~ OH iBU2AlH,CH2Cl2 DMTO ~ ~ r COOEt TESO' ~ 'NHAc TESO' ~ 'NHAc OTES OTES
~ 1. Et3SiCl, irnid., DMF 88 OCH2CH2CN ~CCOOH, CH3NO2, THF

Cl NiPr Pr2EtN, CH2Cl2 OCH2CH2CN

DMTO ~O~l ~O N Pr2 DMTO ~ OTES

TESO' ~ 'NHAc TESO' ~J 'NHAc OTES OTES

Scheme ~. Synthesis of N-acetylglllcos~mine-derived C-glycoside (CG) Scheme 8 summarizes the synthesis of hexamer 116, i.e.
glucose-glucosamine hetero carbonucleotoid (CND).

SUBSTITUTE SH EET (RULE 26) W096127379 PCTrUS96J03227 U ~ ~ ~ O ~ ~

o ~ ~ ~ C ~ ~ J ~ ~ ~ Z O llJ
1~ " O=~L--~ U ~ ~ ~--I --~ ~ U o=n--C O

1~ 0~0 ~ ~~~ -~ 0~0 O IO O ~1~ 0 0=~--O 0=~--O
0~

o I ~ llJ
O ~ O--~--O
O
- S ~~
0~ o O

x -U~
. _ s~:

" oo SUBSTITUTE SHEET (RULE 26) W 096/27379 PcT/u~ 2~7 ~_ 0~0 ~ ~0 "~ o=c---o o o ~o u~-u ~ - o~
O=l~O ~ ~ ~

~ o=~--o o=~--o ¢o=~L--o o o ~0 ~ ; O ~ ~Z~ O_ ~ o I UJ I ~
r I 0=~1--00= 1 --O
O O

CJ Or~--O O \~--0 ~ ¢~ ~ ~

SUBSTITUTE SHEET (RULE 26) WO 96/27379 PCTnUS~v~v3227 The chemistries illustrated in Schemes 5 and 8 for synthesizing heteroh~m~r CPD 74 and heterohexamer CND
116 can also be employed for synthesizing homohexamer CPD's 118 (glucose) and 120 (glucosamine) and homohexamer CND~s 122 (glucose) and 124 (glucosamine).

SUBSTITUTE SH EFT (RULE 26) W 096r27379 PCTrUS96/03227 O O ~1:

~\ I ~\ I
0~0 0~0 o ~ I0=~--0 O=tL--O
C,) ~0 ~ Z 1 ~

0~ 0 0~0 "'IO ~"'I

~ \ I o=~l_o o=~l_o 0~ 0~ 0~0 0~0 o=~_o -- 0=~--0 "
\z -- 2 Z ~2 0 0 C

~_0 =~}O y ~_ = ~_o o = ~ _ O

O ~ ~

~I ~J''~~=g--~ g ~
~ ~
A
o~o o~o O

SUBSTITUTE SH EET (RULE 26) CA 022l4789 l997-09-0~
W 096/27379 PCTrUS9G)v~227 In analogy with the construction o~ oligopeptide and oligonucleotide libraries, a oligosaccharide carbopeptoid (CPD) library may be constructed by per~orming using a split synthesis method o~ oligomerization as illustrated in Scheme 500 for carbopeptoids and Scheme 550 for carbonucleotoids. For example, the split synthesis may employ beads upon which to build the oligomers. Beads are aliquoted into each o~ a several reaction vessels, each reacrtion vessel cont~ini~g a different core molecule. The core molecules are then allowed to attach to the beads. The beads are washed, mixed with one another, and then re-aliquoted (split) into a second set of reaction vessels for addition o~ a second core molecule to the first added core molecule. The process is then reiterated until the oligomerization process is complete. The resultant library of oligosaccharides may then be screened using conventional methods developed ~or oligopeptide and oligonucleotide libraries. Screening an oligosaccharide library can lead to the identification of individual oligosaccharide components within the library having binding activity and/or bioactivity.
The above oligosaccharide libraries (CPD and CND) may be enlarged by introducing additional functionalities into the basic CA's and CG's.
' The above oligosaccharide libraries (CPD and CND) may be further enlarged by enlarging the pool of free functional groups on the CA's and CG's and employed this enlarged pools of CA's and CG's during the respective split synthesis processes.

SU BSTITUTE SH EET (RU LE 26) W096/27379 PCT/u~r~32~7 Scheme 20 illustrate a protocol published by Fuchs, E.F. et al. (~. Chem Ber. 1975, 108, 2254) for the synthesis of CA 45 and 46 from glucose pentaacetate.
Additionally, Scheme 20 illustrates a synthetic route for CG 82, also starting ~rom glucose pentaacetate The reagents and conditions for synthesizing CG 82 are provided as follows:

Steps (a)-(d): according to Fuchs (supra).
Step (e): (1) DMTCl, DMAP, Pyridine; room temperature.
(2) TESTfl; 0~C.
Step (f): DIBAL-H,=CH2Cl2; -78~C; and Step (g): (NCCH2CH2)(NiPr2)PCl, tetrazole, CH2Cl2-The reagents and conditions for synthesizing CA 46 from CA 45 are provided in Step M as follows:
Step (m): FMOC-Cl, K2CO3, THF, H2O; 0~C.

SUBSTITUTE SHEET (RULE 26) W 096/27379 ~-lIU~_5.'~3227 AcO ~ o ~ OAc HO ~ ~ ~ COOMe DMTO ~ O ~ COOMe AcO' ~ 'OAc HO' ~ "OH TESO' ~ 'OTES
OAcOH OTES
36~ 76 78 RNH~O CO2H ~o TESO' ~OTESDMTO ~ O~ ~ ~O NiPr2 DMTO ~O~ 'OH
45OTEHTESO' ~ "OTES ~ TESO' ~ 'OTES
r r ~ OTES OTES
~146: R=FMOC ~ ~ 80 Scheme 20. Conventional route to CAs and a variation for the synthesis of CG. Reagents and conditions: (a) - (d) corresp. Lit.: (e) (1) DMTCl, DMAP, Py; RT. (2) TESTfl; oor (f) DIBAL-H, CH2Cl2; -78 ~C. (g) (NCCH2CH2)(NzPr2)PCl, tetrazole CH~C12. (m~ FMOC-Cl, K2CO3, l~IF, H20; 0 ~C.

SUBSTITUTE SHEET (RULE 26) W 096/27379 PCTrUS96/03227 A synthetic route for producing C-glycosides (CG's) with B-configuration at the former anomer center is illustrated in Scheme 21. The starting material (compound 36) is commercially available. The reagents and conditions for synthesizing CG 181 and CG 185 are as follows:
Step (a): Co2(CO)8, HSiEt2Me, CO.
Step (b): (1) AcOH, H20, THF;
(2) RuC13, NalO4, CH3CN, H20, CCL4, room temperature;
Step (c): NaOMe, MeOH;
Step (d): (1) DMTCl, DMAP, Pyridine, room temperature;
(2) TESOTf;
Step (e): BH3-THF;
Step (f): (NCCH2CH2)(NiPr2)PCl, tetrazole, Ch2C12 i Step (g): (1) 1 equiv TsCl. base (2) TESOTf;
Step (h): NaN3;
Step (i): H2, Pd(OH)2-C;
Step (j): FMOC-Cl, base.

SUBSTITUTE SHEET (RULE 26) W 096127379 ~CTSUS96S032~7 AeO--~O~OACAcO~ ~1--OSiMe3 b ACO ~O~CO2H
AeO' ~ 'OAeAcO~ ~ 'R AeO' ~ 'R
OAe OAe OAe R = OAc or NPhth or NHAc c N ~ O

DMTO f ~~-- NiPr2 DMTO~O~--OH HO ~,O~CO2H
TESO' ~J 'R ~ TESO~ ~IJ 'R d,e HO' ~J 'R
OTES OTES OH
181 ~ 180 178 R = OTES or NPhth or NHAc R = OAc or NPhth or NHAc FMOCNH 1~~ CO2H N3~O~CO2H TsO I~O~CO2H
TESO' ~R TESo' ~ 'R TESO' ~ 'R
OTES OTES OTES

185 R = OTES or NPhth or NHAc Scheme 21. Synthesis of C-glycosides with ~-configuration at the former ,~nomPric center. Re~lgents and conditions: (a) Co2(CO)8, HSiEt2Me, CO. (b) (1) AcOH, H20, THF; (2) Rua3, NaIO4, CH3CN, H20, CCl4, RT. (c) NaOMe, MeOH. (d) (1) DMTCl, DMAP, Py, RT: (2) TESOTF. (e) BH3-THF. (fl (NCCH2CH3)(NiPr~)PCl, tetrazole, CH2Cl2. (g) (1) 1 equiv TsCl, base: (2) TESOTf. (H) NaN3. (i) H~, Pd(OH)2-C. (j) FMOC-Cl, base.

SUBSTITUTE SHEET (RULE 26~

W O 96/27379 PC~rrUS96/03227 BnO ~ .~ ~ a-b TsO ~ ~ ~ COzH
BnO' 'OBn 191-192 TESO' 'OTES TESO' 'OTES
OBn OTES OTES

a,~,h,i ef FMOCNH ~ O ~CO2H
DMTO ~ ~ ' ~ OH HO2C ~ ~ ' ~ NHFMOCTESO' ~ 'OTES
TESO' ~ 'OTES 201-204 TESO' ~ 'OTES OTES
OTES OTES
200 ~

Scheme 22. Svnthesis of C-glycosides with a-configuration at the former anomeric center. Reage~2ts nt2d CO?lditiO?25: (a) reductive debenzylation. (b) (1) 1 equiv TsC1, base; (2) TESOTf. (c) NaN3. (d) RuCl3, NaIO4, CH3CN, H20, CCl4.
(e) H2, Pd-C. (f) FMOC-Cl, base. (g) (1) DMTC1, DMAP, Py, RT; (2) TESOTf.
(h) (1) RuC13, NaIO4, CH3CN, H~O, CC14; (2) CH~N2. (i) DIBAL-H. (j) PPh3, DIAD, diphenyl phosphoryl azide (DPPA), THF. (k) KMnO4, t-BuOH, buffer.

SUBSTITUTE SHEET (RULE 26) W 096127379 P~-llu~ 3227 Synthetic routes for producing with C-glycosides with a-con~igurations at the ~ormer anomeric center, i.e.
CG 196 and CG 1204, are illustrated in Scheme 22. The common starting material for these synthetic routes (compound 190) is disclosed by Schmidt, R. R. et al.
(Liebigs Ann. Chem. 1987, 825). The reagents and conditions for the reactions lea~ing to CG 196 and CG 204 are as follows:
Step (a): reductive debenzylation;
Step (b): (1) e~uiv TsCl. base;
(2) TESOTf.
Step (c): NaN3.
Step (d): RuCl3, NalO4, CH3CN, H20, CCl4.
Step (e): H2, Pd-C.
Step (f): FMOC-Cl, base.
Step (g): (1) DMTCl, DMAP, Pyridine, room temperature;
(2) TESOTf.
Step (h): (1) RuCl3, NalO4, CH3CN, H20, CC14;
(~) CH2N2 Step (i): DI3AL-H.
Step (j): PPh3, DIAD, diphenyl phosphoryl azide (DPPA),THF.
Step (k): KMnO4, t-BuOH, buf~er.
Reactions for the development of the galactose derived C-glycoside 138 into protected CA's and diols is illustrate in Scheme 23. The common starting material for these synthetic routes (compound 138) is disclosed by SUBSTITUTE SHEET (RULE 26) W O 96127379 PC~rrUS96/03227 Petrus, L. et al. (Chem. zvesti. 1982, 3 6, 103) . The reagents and conditions required for the synthesis of compound 209, compound 2 14, compound 220, and compound 224 are indicated below:
Step (a): (1) 1.1 e~uivalent DMTCl, DMAP, Pyridine, 12 hour, 20~C;
(2) TesOTf, CH2, 0~C, 1 hour, 83%.
Step (b): (1) LAH, ether, reflux, 2 hour;
(2)FMOC-Cl, K2CO3, THF, H2O, 0~C, 1 hour, 55%;
Step (c): 10% HCOOH in CH2Cl2, 0~C, 2 minutes, 100%.
Step (d): RuCl3, NalO4, CH3CN, H2O, CCl4, 20~C, 10 minutes, 54%.
Step (e~: (1) 1 equiv. TsCl, base;
(2) TESOTf.
Step (f): NaN3.
Step (g): oxidative NEF.
Step (h): Pd-C, H2.
Step (i): FMOC-Cl, base.
Step (j): (1) 1 equiv. PivCl, base;
(2) TESOTf.
Step (k): (1) oxidative Nef; (2) CH2N2.
Step (l): DI~AL-H.
Step (m): DMTCl, DMAP, Pyridine.
Step (n): LAH.
Step (o): Nef reaction Step (p): LAH.

SUBSTITUTE SH EET ~RULE 26) WO 96/27379 PCI)US96/03227 HO2C~O~f N(H)FMOC a-d i FMOCNH l~orc02H

TESO~ 'OTES 206-209 ¦ ¦ 210-~14 TESO~ 'OTES
OTES O OTES
[~ HO~ ~ NO
HO~ 'OH
OH

DMTO~ ~--OH l l HO/~ ~ODMT
TESO~ 'OTES j-n TESO~ 'OTES
OTES a, o, p 215-2Z0 OTES
~ I

Scheme 23. Development of the galactose derived C-glycoside 138 to protected CAs and diols. Reagents and conditions: (a) (1) 1.1 equiv Dl~TCl, DMAP, Py, 12h, 20 ~C; (2) TesOTf, CH2Cl2, 0 ~C, lh, 83%. (b) (1) LAH, ether, reflux, 2h; (2) FMOC-Cl, K2C03, THF, H20, 0 ~C, lh, 55%. (c) 10% HCOOH ~n CH2Cl2 0 ~C, 2 min, 100%. (d) RuCl3, NaIO4, CH3CN, H2O, CCl4, 20 ~C, 10 min, 54%. (e) (1) 1 equiv TsCl, base; (2) TESOTf. (f) NaN3. (g) oxidative Nef. (h) Pd-C, H2. (i) FMOC-Cl, base. (j) (1) 1 equiv PivCl, base; (2) TESOTf. (k) (1) oxidative Nef; (2) CH2N2. (l) DIBAL-H. (m) DMTCl, DMAP, Py. (n) LAH. (o) Nef reac~ion. (p) LAH

SUBSTITUTE SHEET (RULE 26) W 096/27379 PCTrUS9GI'~32~7 An exemplary protocol ~or synthesizing a hexamer carbopeptoid (CPD 234) starting from galactose derived CA
214, glucosamine derived CA 62, and glucose derived CA, using standard methods ~or solid phase peptide synthesis is illustrated in Scheme 24 The reagents and condition ~or these reactions are as ~ollows:
Step 1: DCC, HOBT, Et3, DMF;
Step 2: Piperidine, DMF

SUBSTITUTE SHEFT (RULE 26) CA 022l4789 l997-09-05 W 096127379 PCTnUS9~03227 Q -~

O _ f O
~_ Cl (~3 ~ G ~ Z G
0~

I I

,_ @ o G

~ C ~
0~0 ~,:, 0~0 O G
C ~, ) Z O ~

,.

SUBSTITUTE SHEFT (RULE 26) W096/27379 P~lIU~,'/03227 Q ~ ~
O--/ ~ O--/ ~ 0=/ ~~ ~,~
c ~Z =~ ~ ~ 0~ ~

0~ ~0 0=~,~0 ==~
or~

cO~ o ~ 0~ < " ~ I ~;

~0 G
I

SUBSTITUTE SHEET (RULE 26) , W096~7379 ~lJU~J~3227 n~ lC D~ilnOL~S

Preparation of 37 A ~--1'~' ~C N

AcO' ~ OAc AcO

To a solution of ,B-D-Glucose pentaacetate 36 i n nitromethane from Aldrich company (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (lX), water (lX), brine (lX) and then dried over ma nesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 37 as a white solid (47%). scheme 3 step 1; scheme 9, step a.

SUBSTITUTE SH EET (RU LE 26) W096~7379 r~l/u~-5~o3227 Preparation of 38 HO--l--O~COOEt HO' (~ 'OH
OH

The crude product 37 is next dissolved in ethanol (0.15 M) and then concentrated H2S O4 (0.01 equivalents-catalytic) is added. The reaction mixture is heated to 85 ~C for eight hours.
The solution is next concentrated in vacuo and purification by 10flash column chromatography affords compound 38. scheme 3 step 2 Preparation of 39 PivO~o~cooE~
HO' ~ OH
OH

To a solution of 38 (1.0 equivalents) in pyridine (.10 Molar), is added trimethylacetyl chloride (pivaloyl chloride) (2.5 20equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium SUBSTITUTE StlEET tRULE Z6) W 096~ m 9 ~-llU' ~CJD3227 chloride (2X), copper sulfate (2X), brine ( lX), dried over MgSO4 and concentrated. Purification by flash column chromatography affords compound 39. scheme 3 step 1 Preparation of 40 PivO~T~O~COOEt TESO' ~ OTES
OTES

To a solution of 39 (1.0 equivalents) in methylene chloride (.10 Molar), is added diisopropylethylamine (3.3 equivalents) at 0 ~C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 40. scheme 3 step 2 SUBSTITUTE SHEET (RULF 26) W096~7379 ~ 3??7 Preparation of 41 HO--~O~COOEt TESO' ~ 'OTES
OTES
41.
s To a solution of 40 in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column chromatography affords compound 41. scheme 3 step 1 Preparation of 42 N3~0~COOEt TESO' ~ 'OTES
OTES

A solution of 41 (l.0 equivalents) in tetrahydrofuran (.18 M) is treated with DPPA (diphenylphosphorylazide, 2.0 equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD
(diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is heated to 80 ~C for 3 hours and then diluted with ether (2X) and SUBSTITUTE StlEET (RULE 26) W 096/27379 ~l~U'_ÇJ~

washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgSO4 and evaporated. Purification by flash column chromatography affords compound 42. scheme 3 step 2 Preparation of 44 H2N ~O~,~COOEt TESO' ~ 'OTES
OTES

10A solution of 42 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 ~C. The mixture is next exposed to 10% Pd/C
(.1 equivalents) and is then subsequently capped with a hydrogen balloon at l atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is 15subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 44. scheme 3 step 1 SUBSTITUTE SHE~T (RULE 26) CA 02214789 1997-09-0~
wog6n7379 ~/u.~'~03227 Preparation of 45 2 ~~ r TESO" ~ 'OTES
OTES

s A solution of 4 4 ( 1.0 equivalents) is dissolved in p-dioxanes (.1 M) and then exposed to a solution 3.0 Molar solution of sodium hydroxide ( 1.5 equivalents). The reaction is then stirred for 2 hours at 50 ~C and is subsequently diluted with ether and washed with a solution of NH4Cl (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 45. scheme 3 step Preparation of 46 To a solution of 45 (1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried ( M g S O 4 ) and concentrated. Purification by flash column SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
wo s6/2ms ~ V~ ?7 chromatography affords compound 46. scheme 3 step 2 Preparation of 48 r ACo--~o ~OAc AcO' ~ ' NPhth OAc Procedure as described in Methods in Carbohydrate chemistry, Whistler, R., II, 19 6 3, p. 327. A mixture of 80g anhydrous D-glucosamine hydrochloride or D-galactosamine hydrochloride from Aldrich chemical company, in 200 mL.
methanol and 20g Dowex 50 (H+) acidic resin, is stirred at the boiling point in a round bottom flask. After 24-hr. reaction time, the resin is removed by filtration and ished three times with 20 ml. of methanol. The filrate and ishings are combined and concentrated to about 125 ml by rotovap. The concentrate is allowed to cool to room temperature and the product crystallizes overnight.
To a solution of free amine, in chloroform (.5 M), is added phthalic anhydride ( 1.5 equiv.) and the reaction mixture is allowed to reflux at 70 ~C for 4 h. The product is then crystallized and carried onto the next step.
To a solution of triol in methylene chloride (.5 M), is added SUBSTITUTE SHEET (RULE 26) wog6r27379 ~ 1u~,5~'03n7 acetic anhydride (3.5 equiv.) and triethyl amine (3.5 equiv.) and the reaction mixture is allowed to stir at O ~C for 4 h. The product 4 8, is then crystallized or purified by flash column chromatography and carried onto the next step.

SUBSTITUTE StlEET (RULE 26) CA 022l4789 l997-09-0~
W 096/27379 1~-~J~6JD3227 Preparation of 50 Aco--~~~C N
AcO' ~ ' NPhth AcO

S To a solution of N-phthalamido-D-Glucosamine tetraacetae 48 in nitromethane (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the rem~ining trimethylsilyl cyanide. The mixture is evaporated and the rem~ining oil is resuspended in dichloromethane and washed with sodium acetate solution ( 1 X), water ( 1 X), brine ( 1 X) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 51) as a white solid (47%).
scheme 4 Preparation of 52 The crude product 50 is next dissolved in ethanol (0.15 M) and then concentrated H2S O4 (0.01 equivalents-catalytic) is added. The reaction mixture is heated to 85 ~C for eight hours.
,~ The solution is next concentrated in vacuo and purification by SUBSTITUTE SHEET (RULE 26) W 096127379 r~ 6/03Z27 flash column chromatography affords compound 52. scheme 4 Preparation of 54 HO--~O~cooEt HO' ~ NHAc A solution of 52 (1.0 equivalents) is dissolved in methanol (.1 M total). The reaction is then charged with acetic anhydride ( 1.1 equivalents) and is subsequently stirred for 2 hours at 30 ~C. The reaction is next diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated.
Purification by flash column chromatography affords compound 54. scheme 4 Preparation of 55 Pivo - ~o~cooEt HO' ~ NHAc OH

SUBSTITUTE SHEET ~RULE 26) W 0961273n ~ r J '03227 To a solution of 54 ( 1.0 equivalents) in pyridine (.10 Molar), is added trimethylacetylchloride (pivaloyl chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine ( 1 X), dried over MgSO4 and concentrated. Purification by flash column chromatography affords compound 55. schenne 4 Preparation of 56 Pivo - ~o~cooEt TESO' ~ 'NHAc OTES

To a solution of 55 (l.0 equivalents) in methylene chloride (. l0 Molar), is added diisopropylethylamine (2.2 equivalents) at 0 ~C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (2.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 56. scheme 4 SUBSTITUTE SHEET tRULE 26) W096127379 ~-l/U~961o3227 Preparation of 57 HO--~O~rcooEt TESO' ~ 'NHAc OTES

s To a solution of 56 in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column 10chromatography affords compound 57. scheme 4 Preparation of 58 N3~0~COOEt TESO' ~ NHAc OTES

A solution of 57 (1.0 equivalents) in tetrahydrofuran (.18 M) is treated with DPPA (diphenylphosphorylazide, 2.0 equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD
20(diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is SUE3STITUTE StlEET (RULE 26) W 096127379 ~-lIU~ 03227 heated to 80 ~C for 3 hours and then diluted with ether (2X) and washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgS O4 and evaporated. Purification by flash column chromatography affords compound 58. scheme 4 Preparation of 60 H2N ~OrCOOEt TESO' ~ NHAc OTES

A solution of 58 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 ~C. The mixture is next exposed to 10% Pd/C
(.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atm. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 60. scheme 4 SUBSTITUTE SH EET (RU LE 26~

WO 96/27379 ~- -lIU~;~ "03227 Preparation of 61 H2N ~~~,C O O H

TESO' ~ NHAc OTES

A solution of 6 0 ( 1.0 equivalents) is dissolved in p-dioxanes (.1 M) and then exposed to a solution 3.0 Molar solution of sodium hydroxide ( 1.5 equivalents). The reaction is then stirred for 2 hours at 50 ~C and is subsequently diluted with ether and washed with a solution of NH4Cl (3X), brine ( 1 X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 61. scheme 4 Preparaffon of 62 H

FMocN~~O~,~COOH
TESO' ~ 'NHAc OTES

To a solution of 61 (1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0 SUBSTITUTE SHEET (RULE 26) W096/27379 ~ lU~61o3227 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2~), brine ( lX) and then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography affords compound 62. scheme 4 Preparation of 63 FMOCN--~~~ 11 N ro~1~COOEt TESO"' ~/ 'OTESTESO ~/ 'NHAc OTES OTES

To a stirred solution of the acid 46 ( 1.0 equivalents) and the amine 6 0 ( 1 .1 equivalents) in dimethylformamide ( .10 Molar) at 25 ~C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Next dicyclohexylcarbodiimide (1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 6 3 .

SUBSTITUTE SHEET (RULE 26) W096~7379 ~1/U~6~'~32>7 scheme 5 step 1 Preparation of 64 H2N~ ~ I N--~o~COOEt TESO"'y 'OTESTESO ~ 'NHAc OTES OTES

To a stirred solution of 63 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents) . The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CuS O4 (2 X)7 water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 64. scheme 5 step 2 Preparation of 65 FMOCN~O~ N--~COOEt TESO' ' NHAc 'OTESTESO ' NHAc OTES OTES OTES

SUBSTITUTE SHEET (RULE 26) wos~nn ~1J~ D3227 To a stirred solution of the acid 62 ( 1.0 equivalents) and the amine 64 (1.1 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using S the acid 62 or intermixing with other acids including for example acid 46 to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme ~ ) to obtai7l large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours.
The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X).
The organic phase is dried over MgSO4 and then concentrated.
Purification by flash column chromatography affords compound 65. scheme 5 step 1 Preparation of 66 NH2 ~~ ll N/~O-- ll N l o~COOEt ESO '~J OTESTESO"'~"""N
OTES OTES OTES

To a stirred solution of 65 ( 1.0 equivalents) in SUBSTITUTE 5HEET (RULE 26) W 096~7379 r~~ o3227 dimethylformamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents) . The reaction is stirred for 1 hour and is then ~, diluted with ether, and washed with aqueous CuSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 66. Note: numerous iterations can be performed using variable length oligomers of 66 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme S). scheme ~ step 2 Preparation of 67 o o o MOCN--~O-- ll N~O-- --N--~O~ ~ N~O~cOoEt --~ TESO ~ 'NHAC TEso~--~J 'OTESTEso~ NHA
OTES OTES OTES OTES

To a stirred solution of the acid 46 (1 0 equivalents) and the amine 66 (1~1 equivalents) in dimethylformamide ( 10 Molar) at 25 ~C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 46 or intermixing with other acids including for example acid 62, to form successive oligomers where n=2 to infinit~ (a hexamer is shown in scheme 5 ) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 SUBSTITUTE S~EET (RULE 26) wa 96 m 379 ~-lJU~Jv3227 equivalents) is added and the reaction is stirred for 14 hours.
The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X).
The organic phase is dried over MgSO4 and then concentrated.
Purification by flash column chromatography affords compound 67. scheme 5 step 1 SUBSTITUTE SHEET (RULE 26) W096~7379 ~l/U~Ç~'03Z27 Preparation of 68 "
o o ,, H2N--~ H ~ --N--~o~ rl N l O~COOEt TESO ',J OTESTESO' ~ '''NHAcTESO'' 'lJ 'OTESTEso~"~ "'NHA

To a stirred solution of 67 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents) . The reaction is stirred for l hour and is then diluted with ether, and washed with aqueous CuSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 68. Note: numerous iterations can be performed using variable length oligomers of 68 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme ~ step 2 Preparation of 69 H ~ 1I N- ~ ~r ~ N ~ 1I N ~ COOEt TESO' 'NHAc 'OTESTESO' 'NHAcTESO "OTESTESO' 'NHAc OTES OTES OTES OTES OTES

A
To a stirred solution of the acid 62 ( 1.0 equivalents) and SUBSTtTUTE SH EET (RULE 26) CA 02214789 1997-09-OF, WO 96127379 ~-llU' ~ 13~7 the amine 6 8 ( 1.1 equivalents) in dimethylformamide (.10 ~, Molar) at 25 ~C, is added l-hydroxybenzotriazole (HOBT; 1.1 equivalents). No~e: numerous iterations can be performed using the acid 62, or intermixing with other acids including for example acid 46, to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours.
The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X).
The organic phase is dried over MgSO4 and then concentrated.
Purification by flash column chromatography affords compound 69. scheme 5 step 1 .

Preparation of 70 O o o o ~2N~~~L1~ 1 H--~ ~N ~O~N--~O~COOEt y TESff--~ 'OTESTESO" ~

To a stirred solution of 69 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents). The reaction is stirred for 1 hour and is then SUBSTITUTE SHEET(RUI_E 26) W 096~7379 ~ 5f'~ 7 diluted with ether, and washed with aqueous CuSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 70. Note: numerous iterations can be performed using variable length oligomers of 70 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme 5 step 2 Preparation of 71 FMOCN ~~~ ~ N ~ i--N--~r~~r~ O~COOEt TESO~y ~OTE~SO 'NHAcTESO" ~OTESTESO~ 'NHAcESO (,~lt~>lt~7C/~ ~ 'NHAc OTES OTES OTES OTES OTES OTES

To a stirred solution of the acid 46 ( 1.0 equivalents) and the amine 70 (1.1 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added l-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 46 or intermixing with other acids including for example acid 62, to form successive oligomers where n=2 to infirzity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours.
The mixture is diluted with ether, filtered and the filtrate is SUBSTITUTE SH EET (RULE 26~

WO961273n ~ U~_.'/03227 washed with aqueous NaHC03 (2X), water (2X), and brine (2X).
The organic phase is dried over MgS04 and then concentrated.
Purification by flash column chromatography affords compound 71. scheme 5 step 1 SUBSTITUTE SHEET (RULE 26) W096/27379 r~ ',5~03227 Preparation of 72 H2N~~N~--~N--~, r~ L N~ ~-- N ,~COOEt TESO' ""~OT~SO 'NHAcTESO" OTES TESO' 'NHAcESO ~ t~ t~ ' NHAc OTES OTES OTES OTES OTES OTES
s To a stirred solution of 71 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents) . The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CuSO4 (2X), water (2X)~ and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 72. Note: numerous iterations can be performed using variable length oligomers of 72 to form peptoid oligomers where ~=2 to infinity (a hexamer is shown in scheme 5). scheme ~ step 2 Preparation of 74 H2N r~O~ ~LNH ~ ~ N--~O~ O~N ~O~COOH
,, H H "'NHA HO'" '~; HO' ~ 'OH ~ OH HO' ~ 'NHAC

To a stirred solution of 72 ( 1.0 equivalents) in acetonitrile SUBSTITUTE SHEET (RULE 26) W096~7379 r~l/u~5~o3227 (.50 Molar) is added an HF pyridine solution (.50 M) from Aldrich chemical company. The reaction is allowed to stir for five hours and is then condensed. The crude 73 oligomer is then resuspended in p-dioxane (.50 Molar) to which is added a 3.0 Molar solution of NaOH (3.0 equivalents). The reaction is stirred for 1 hour at 50 ~C and is then quenched with aqueous NH4Cl (2X) and subsequently lyophilized. Purification by HPLC
chromatography affords compound 74. scheme Preparation of 76 HO--I~OvcOOEt HO' ~ 'OH
OH

To a solution of ,B-D-Glucose pentaacetate 36 i n nitromethane from Aldrich company (.13 Molar), is added trimethylsilylcyanide (3.0 equivalents) and then tin tetrachloride (.02 equivalents). Note: other pyranose sugars such as ,B-D-Mannose, ,B-D-Galactose pentaacetate and other lewis acids such as BF30Et~ may be used for alternative derivatives. The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the SU85TITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
W096/27379 ~-llU~r~r~7 rem~ining trimethylsilylcyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (lX), water (lX), brine (lX) and then dried over magnesium sulphate and concentrated. The crude product is next dissolved in ethanol (or methanol if the O-methyl glycoside is desired as in scheme 20), (0.15 M) and then concentrated H2 S O 4 (0.01 equivalents) is added. The reaction mixture is heated to 85 ~C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography affords compound 76. scheme 6; 76, scheme 20 (as the O-methyl glycoside).

Preparation of 78 ~MTO~ COOEt TESO' ~ 'OTES
OTES

To Tetrol 7 6 ( 1.0 equivalents) in pyridine ( .10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX) dried over MgSO4 and concentrated.

SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096/~7379 ~ ,.S'03227 Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (4.4 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (4.4 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 78, scheme 6; 78, scheme 20 (as the O-methyl glycoside).

Preparation of 80 ~MTO~ OH
TESO' ~ 'OTES
OTES

To a solution of 78 (1.0 equivalents) in methylene chloride (.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X)~
brine (lX) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords ..

SU~STITUTE SHEET (RULF 26) W096~7379 r~liU~3G/03227 compound 80. scheme 6 Preparation of 82 N(~Pr)2 ~MTO--~O O'P'O~--C N
TESO' ~ 'OTES
OTES

To a solution of 80 (1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nucl.
Acids Res. 1984, 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( lX) and is then dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 82 (66~o yield).
scheme 6 SUBSTITUTE SHEET (RULE 26) W096/27379 ~ 961n3227 Preparation of 84 ,.
H O~,l' 1~ OTFS
TESO' ~ 'OTES
OTES

To 80 (1.0 equivalents) in methylene chloride (.10 Molar) at 0 ~C, is added diisopropylethylamine ( 1.1 equivalents).
Subsequent addition of triethylsilyl trifluoromethanesulfonate (1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. The crude is then resuspended in nitromethane and exposed to 10% Cl3COOH (1.1 equivalents) in THF (.10 Molar). The reaction is stirred at 0 ~C for 2 hours and is then diluted with ether and washed with sodium bicarbonate (2X), brine ( lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 8 4 .
scheme 6 SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W 096/27379 ~-1/U~_5J03227 Preparation of 86 ~, Ho--~OrCO~Et HO' \~/ 'NHAc OH

5 To a solution of N-phthalamido-D-Glucosamine tetraacetate 48 in nitromethane (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the rem~ining oil is resuspended in dichloromethane and washed with sodium acetate solution (lX), water (lX), brine (lX) and then dried over magnesium sulphate and concentrated. The crude product is next dissolved in ethanol (0.15 M) and then concentrated H~SO4 (0.04 equivalents) is added. The reaction mixture is heated to 85 ~C for eight hours. The solution is next concentrated in vacuo and is then resuspended in methanol (.10 M) and acetic anhydride ( 1.1 equivalents) from Aldrich company is added in one step. After 2 hours, condensation and purification by flash column chromatography affords compound 86. scheme 7 SU BSTITUTE S H EET (RU LE 26) W~96~7379 ~ ,56J0322 Preparation of 88 )MTO--I' ~COOEt TESO' ~ NHAc OTES

To Triol 86 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine ( lX), dried over MgSO4 and concentrated. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 88. scheme 7 SUBSTITUTE SHEET (RULE 26) W 096/27379 ~1/US96103227 Preparation of 90 ~MTO~ OH
TESO~ ~ 'NHAc OTES
To a solution of 88 (1.0 equivalents) in methylene chloride (.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X), brine (lX) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 90. scheme 7 Preparation of 92 N(~Pr)2 ~MTO~ ~~O' O~
TESO' ~ 'NHAc OTES

To a solution of 90 ( 1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 SUBSTITUTE SHEET tRULE 26) CA 02214789 1997-09-0~
W096~7379 ~-lJU~,5Jo3227 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nucl.
Acids Res. 1984, 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( lX) and is then dried (MgS04 ) and concentrated.
Purification by llash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 92 (66% yield).
scheme 7 Preparation of 94 HO~ '~--OTES
TESO' ~ 'NHAc OTES

To 90 (1.0 equivalents) in methylene chloride (.10 Molar) at O ~C, is added diisopropylethylamine ( 1.1 equivalents).
Subsequent addition of triethylsilyl trifluoromethanesulfonate ( 1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried (MgS04) and concentrated. The crude is then resuspended in nitromethane and exposed to 10% Cl3COOH (1.1 equivalents) in THF (.lO
Molar). The reaction is stirred at O ~C for 2 hours and is then SUBSTITUTE SHEET (RUEE 26) CA 02214789 1997-09-0~
W 0961~7379 ~-1/U'-.,"03227 diluted with ether and washed with sodium bicarbonate (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 9 4 .
scheme 7 Preparation of 98 (homodimer scheme 8) To a solution of 94 (1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 ~C. Next, a solution of 82 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and I2 (4.0 e~uivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine ( 1 X) and dried (MgSO4) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1:1), (.01 M) and stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 98 (scheme 8).

SUBSTITUTE SHEET ~RULE 263 CA 02214789 1997-09-0~
W096~7379 ~ ,Cl03227 Preparation of 102 (heterotrimer scheme 8) To a solution of 9 8 ( 1 .0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 ~C. Next, a solution of 92 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine ( 1 X) and dried (MgSO4) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1: 1 ), (.01 M) and stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 102 (scheme 8).

Preparation of 106 (heterotetrarner scheme 8) To a solution of 102 (1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 ~C. Next, a solution of 8 2 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096~7379 ~1/U'~ 3227 with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine (lX) and dried (MgSO4) and concentrated.
Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3:1:1), (.01 M) and stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 106 (scheme 8).

Preparation of 110 (heteropentamer scheme 8) To a solution of 106 (1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company (10.0 equivalents) at 25 ~C. Next, a solution of 92 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an SUBSTITUTE SHEET (RULE 26) .

CA 02214789 1997-09-0~
WO 96/273'79 ~ u.,~OES03227 additional 5 minutes and is next diluted with ether and washed , with brine ( 1 X) and dried (MgS04 ) and concentrated.
Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3:1:1), (.01 M) and stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHC03 (3X), brine (lX) and dried (MgS04) and concentrated. Purification by flash column chromatography affords compound 110 (scheme 8).

Preparation of 114 (heterohexarner scheme 8) To a solution of 110 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 ~C. Next, a solution of ~ 2 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine (lX) and dried (MgS04) and concentrated.
Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3 :1:1), (.01 M) and stirred for 18 hours at 25 ~C. The reaction is SUE~STITUTE S~tEET (RULE 26) CA 02214789 1997-09-0~
W09612 m 9 ~-1/U~6/03227 then diluted with ether and washed with NaHCO3 (3X), brine (1~) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 114 (scheme 8).

Preparation of 116 (heterohexamer scheme 8) To a solution of 114 (1.0 equivalents) in methylene chloride (.10 M), is added a solution of HF-pyridine ( 1.0 M) at O
~C. The reaction is next stirred for an additional 30 minlltes and is next diluted with ether and washed with a saturated solution of sodium bicarbonate (3X), copper sulfate solution to remove the pyridine (2X) brine (lX), dried (MgS04) and concentrated.
Purification by flash column chromatography and then the product is resuspended in concentrated aqueous ammonium hydroxide and acetonitrile ( 1:1), (.1 M total). The reaction is then stirred for 2 hours at 50 ~C and is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 116 scheme 8.

SUBSTITUTE SHEET (RULE 26) ~ JU~.~~'03227 --8~ -ACo~r~r~AC a - d HO--rOrCOOMee g Bu~Me2SiO~rOl~OH
AcO"y 'OAc (126-130)HO' ~ 'OH (131-134) BnO''y 'OBn OAc OH OBn cl~P o~CN~ N(~Pr)2 1 0 Bu~Me2SiO~rO'I~OP O~CN HO ,O~
BnO' ~ 'OBnBnO'' y "OBn OBn OBn Ii R10--rO'I--O-P-O--~O~--OR
R20' y 'OR2 R20 Y oR2 k 140: R1 = SitBuMe2; R2 = Bn o C 142: R1 = H; R2 = Bn; R4 = N~3 ~li R o~~~o OpRo~o~ ~R3 ~o~
R20' ~ 'OR2 ~ R20' Y 'OR2 ~ RZO' Y ~oR2 k 144: R1 = SitBuMe2; R2 = Bn; R4 = Nphth C146 R1 = H; R2 = Bn; R4 = Nphth E~i R10 ~ OR3,~,o~ 0 OpRo3 ~0~ 0 p 0~~~~l~OR4 R2~'0R2 ~ R20 y 'OR2 ~ R20' Y 'OR2 ~ R20' Y ~oR2 k 148: R1 = Si~BuMe2; R2 = Bn; R3 = CH2CH2CN; R4 = Nphth 150 R1 = H; R2 = Bn; R3 = CH2CH2CN; R4 = Nphth _ 152: R1 = R3 = H; R2 = Bn; R4 = Nphth mL- 154: R1 = R2 = R3 = H; R4 = Nphth Scheme 9. Synthesis of a Carbonucleotoid SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0=.
W096/27379 ~ G~227 Preparation of 125 A O ~O~C N
AcO' ~ 'OAc OAc To a solution of ,~-D-Glucose pentaacetate in nitromethane from Aldrich company (.13 Molar), is added trimethylsilylcyanide (3.0 equivalents) and then borontrifluoride etherate (.02 equivalents). Note: other pyranose sugars such as ~-D-Mannose, ,B-D-Galactose 0 pentaacetate and other lewis acids such as SnCI4, may be used for alternative derivatives. The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilylcyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution ( lX), water ( lX), brine ( lX) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 125 (also 37) as a white solid (47%). scheme 9 step a SUBSTITUTE SHEET (RULE 26) W096~7379 ~ 56J03227 ~ -83-Preparation of 126 Ho ~O~C N

H O' ~ O H
OH

5To a solution of 12 5 in methanol (.13 Molar), is added sodium methoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The dark brown solution is then concentrated in vacuo to give a dark brown syrup of compound 126 which is carried on without purification 10as a crude oil for the next step. scheme 9 step b Preparation of 127 HO--rO~COOH
HO' ~,J 'OH
OH

The crude product 126 is dissolved in 25% NaOH (0.5 M) and heated at reflux for 18 hours (vigorous reflux is necessary).
Next, the solution is diluted with an addition of water (0.1 M) and to this solution is added Amberlite 112120 resin (H+-form) 20and is then stirred. The supernatant is then decanted and the SU~STITUTE SH EET (RULE 26) W096127379 ~-~/U~,''03227 resin is washed until the eluate is colorless. The eluate is then collected, condensed and azeotroped with MeOH which yields 127 as a crude, pale yellow syrup (47%).

Preparation of 130 HO~l~O~cOoMe HO' ~ 'OH
OH

The crude product 127 is next dissolved in methanol (0.15 M) and then concentrated HCl (0.01 equivalents) is added. The reaction mixture is heated to 85 ~C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography (silica, 20% methanol in ethyl acetate), affords compound 130 as a white solid (60% yield). scheme 9 step d Preparation of 131 TBDMso - l~o~cooMe H O' ~ 'O H
OH

SlJBSTITUTE SHEET (RULE 26) W096~7379 ~1J~6JD3227 To a solution of 130 ( 1.0 equivalents) in dimethylformamide (.23 Molar), is added imidazole (2.5 equivalents) at 0 ~C. Subsequent addition of tert-Butyl-dimethylsilylchloride (2.5 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate) affords compound 131 as a white solid (93% yield). scheme 9 step e Note: the molecule can be protected with other primary directing protecting groups such as DMT (dinzethoxytrityl), and TBDPS
tert-butyldiphenlysilyl, etc.

Preparation of 132 ButMe2s jo~r~~C~~Me BnO' ~ 'OBn OBn To a solution of 13 1 (1.0 equivalents) in dimethylformamide (.23 M), is added Ag2O (6.0 equivalents) at 25 ~C. Benzyl bromide (9.0 equivalents) is next added and the SUBSTITUTE 5HEET (RULE 26) CA 02214789 1997-09-0~
W096~7379 ~-lIU~,'/03227 reaction is allowed to stir for 20 hours. The reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 132 (83% yield). scheIne 9 step f Note: the choice of the protecting group ~s relative and the molecule can be protected with other protecting groups at C2, C3, C4, such as PMB (paramethoxvbenzyl), TES (triethvlsilyl), TBS (tertbutvldimethyls~lyl), etc.
Preparation of 134 ButMe2SiO~i ~--OH
BnO' ~ 'OBn OBn To a solution of 132 (l.0 equivalents) in tetrahydrofuran (.08 M), is added diisobutylaluminumhydride (DIBALH) (3.0 equivalents) at 0 ~C. The reaction is stirred for 1 hour and then quenched with methanol and diluted with ether. The reaction is next worked-up with ammonium chloride (2X), brine ( lX) and is then dried (MgS04) and concentrated. Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 134 (66% yield). scheme 9 step g SUBSTITUTE SHEET (RULE 26) w~ s6~2ms ~ u~g6/~3227 Preparation of 136 B~ 'fOB ~3 OBn To a solution of 134 (1 0 equivalents) in pyridine (10.0 equivalents), is added naphthoyl chloride (3.0 equivalents) from Aldrich company (3.0 equivalents) at 25 ~C. The reaction is stirred for 45 minutes and then diluted with ether and worked-up with a saturated solution of CuSO4 (2X), brine (lX) and is then dried (MgSO4) and concentrated. The crude product is then exposed to acetic acid/tetrahydrofuran/water (3 :1:1 ) at 25 ~C
and allowed to stir for l 5 hours. The reaction is then diluted with ether and worked-up with brine (2X) and is then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 136 (95% yield). Note: alternatively, one could originally protect the C7 position as a DMT (dimethoxytrityl) functionality and protect the Cl position as a TES (triethyl silyl) group.
Subsequent mild acid hydrolysis of the DMT group leads ~o the SIJ~STITUTE SHEET (RULE 26) W096/27379 P~l/u~ 3227 above compound with the TES group at ~he C1 position and a free hydroxvl at the C7 position. scheme 9 step h Preparation of 138 N(~Pr)2 ButMe2SiO~I' ~~O O~~
BnO' ~'OBn OBn To a solution of 134 (1.0 equivalents) in methylene chloride (. lO M), is added diisopropylethylamine (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984, 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( lX) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 138 (66% yield). scheme 9 step i It should be ~l oted that the oligomerization process as shown below in scheme 9, uses the same C-glycoside 138 in an iterative fashior2. The process can be SUBSTITUTE SHEET (RULE 26) Wo 96/27379 ~-llU!,,C~3227 extended however to include a pool of random or ordered C-glycosides as depicted i~ scheme 8.

Preparation of 140 ~ CN O
rBDMSO~ ~--P--0~ ~1 o l~
BnO' ~ 'OBn BnO' ~ 'OBn ~J
OBn OBn To a solution of 13 6 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole ~rom Aldrich company (10.0 equivalents) at 25 ~C. Next, a solution of 138 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and m-chloroperoxybenzoic acid (4.5 equivalents) is added.
The reaction is stirred for an additional S minutes and is next diluted with ether and washed with brine ( 1 X) and dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 140 (97% yield). scheme 9 step j Note the process can iterate as many times as possible to build large carbonucleotide libraries.

SlJBSTITUTE SH EET (RULE 26) CA 02214789 1997-09-0=.
W096~7379 ~ 96103227 Preparation of 142 ~ C N O

H ~--~~~1--~--P--~--1'~~-- ~
BnO' ~ 'OBn BnO' ~'OBn ~J
OBn OBn A solution of 14 0 ( 1.0 equivalents) in acetic acid-tetrahydrofuran-water (3 :1:1), (.01 M) is stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 142 (95% yield).
scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide libraries.

Preparation of 144 o~ CN o~,CN O
rB D M S O~ ~--P--O ~ ~--P--~ ~ ~ J~
BnO' ~ 'OBn BnO' ~ 'OBn BnO' ~ 'OBn ~J
OBn OBn OBn To a solution of 13 8 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company SUBSTITUTE SH EET (RU LE 26) wog6nn79 PCTnUS96)03227 _91 _ ( 10.0 equivalents) at 25 ~C. Next, a solution of 14 2 (3 .0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to O ~C and m-chloroperoxybenzoic acid (4.5 equivalents) is added.
The reaction is stirred for an additional 5 minutes and is next diluted with ether and washed with brine (lX) and dried ( M g S 0 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 144 (97% yield). scheme 9 step j Nole the 0 process can iterate as many times as possible to build large carbonucleotide libraries.

SUBSTITUTE SHEET (RULE 26) W 096/27379 ~ 103227 Preparation of 146 HO ~O--P--o ~~O--P--o~
Bn~ OBn BXJ'OBn BnO' \I~'OBn OBn OBn OBn s A solution of 14 4 ( 1.0 equivalents) in acetic acid-tetrahydrofuran-water (3:1:1), (.01 M total) is stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine ( lX) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 146 (95% yield). scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide libraries.
Preparation of 148 ~CN ~CN ~CN O
-BDMSO--~~~--O--P--0~~~~~0--P--O~~ ~~O--P--0~~~~~o~
BnO' I 'OBn BnO' I 'OBn BnO' ~ 'OBn BnO' ~ 'OBn b~J
OBn OBn OBn OBn To a solution of 13 g ( 1.0 equivalents) in methylene SUBSTITUTE SHFET (RULE 26) W096~7379 1~ 961o3227 -g3-chloride (.10 M), is added l-H-tetrazole from Aldrich company (10.0 equivalents) at 25 ~C. Next, a solution of 146 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 ~C. After 25 minutes, the mixture is cooled to 0 ~C and m-chloroperoxybenzoic acid (4.5 equivalents) is added.
The reaction is stirred for an additional 5 minutes and is next diluted with ether and washed with brine ( 1 X) and dried ( M g S 0 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 148 (97% yield). scheme 9 step j Note the process can iterate as many times as possible to b~ild large carbonucleotide libraries.

Preparation of 150 ~C~ ~ O~CN o ~o~--r-o~~o_r_o~ 1 5 0 A solution of 14 8 ( 1.0 equivalents) in acetic acid-tetrahydrofuran-water (3:1:1), (.01 M total) is stirred for 18 hours at 25 ~C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 150 (95% yield). scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide SUBSTITUTE SHEET (RULE 26) W096127379 ~1/U'~6'03227 libraries.

Preparation of 152 ~--O ~ o OH ~_o ~
onxl ~o~ nO ~ oun 1 5 2 A solution of 15 0 ( 1.0 equivalents) is dissolved in concentrated aqueous ammonium hydroxide and acetonitrile ( 1:1), (.1 M total). The reaction is then stirred for 2 hours at 50 ~C and is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 80% ethyl acetate in petroleum ether) affords compound 152 (88% yield).
scheme 9 step L

Preparation of 154 --~_~~~ O_~ o D "~ U
"~~ "0 ~0 o ~ ~ 1 5 4 A solution of 15 2 ( 1.0 equivalents) is dissolved in a mixture of ethanol-tetrahydrofuran-acetic acid (2:1:1), (.01 M
total) at 25 ~C. The mixture is next exposed to l O~o Pd/C ( 1.0 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is SUBSTtTUTE Slt EET (RULE 26) CA 02214789 1997-09-0~
W096/27379 ~ U~ 7 subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash colurnn chromatography (silica, 100% ethyl acetate in petroleum ether) affords compound 154 (78% yield). scheme 9 step m Preparation of 174 (R group = OTES, NPhth or NHAc) To a solution of tetraacetate derived from 36 or 48 (glucose or glucosamine derived) in methylene chloride (.1 molar) is added a 1.0 molar solution of Co2(CO)g (1.5 equivalents ) in methylene chloride and diethylmethylsilane ( 1.5 equivalents) at 0 ~C. To the stirring reaction mixture, a stream of carbon monoxide is bubbled at l ml per 10 seconds for 30 minutes. The reaction mixture is then quenched with water ( 1.5 equivalents), diluted with ether, washed with sodium bicarbonate (2x), brine (lx) and dried over magnesium sulfate. The crude is purified by column chromatography and affords product 174.

Preparation of 176 (R group = OTES, NPhth or NHAc) To a solution of compound 174 in acetonitrile/water (l:l ratio, .1 molar combined), is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) at 25 ~C and the muddy black mixture is allowed to stir for 1.5 h. The mixture is then diluted with ether (25 mL), washed with water (2X 5 .0 mL) and brine ( lX 5 mL). The SUBSTITUTE SHEET (RULE 26) WO96127379 ~ 32 aqueous layer is back extracted (2X), recombined, and the organic layer was then dried MgS04 and evaporated.
Purification by flash column chromatography yields the desired product 176.
s Preparation of 178 (R group = OTES, NPhth or NHAc) A solution of triacetate 17 6 ( 1.0 equiv.) in methanol (0.5 M), is treated with NaOMe (0.4 equiv.) and allowed to stir at 25 ~C for 24 h. The reaction mixture is then condensed and purified by flash column chromatography to afford compound 178.

Preparation of 180 (R group = OTES, NPhth or NHAc) To triol 178 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) ( 1.~ equivalents) at O
~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgSO4 and concentrated.
Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at O ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgS04) and SUBSTITUTE SHEET (RUI E 26) CA 02214789 1997-09-0~
W~ 96~7379 ~-1r~56/03227 concentrated. Purification by flash column chromatography affords the intermediate acid, which is then resuspended in THF
(1.0 M) and exposed to a 1.0 M solution of BH3-THF (1.5 equivalents) at 0 ~C for 1 hour. The reaction is then quenched with methanol for an additional hour and the crude is then diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried (MgSO4) and concentrated.
Purification by flash column chromatography affords the desired tetraprotected alcohol 180.
Preparation of 181 (R group = OTES, NPhth or NHAc) To a solution of 180 (1.0 equivalents) in methylene chloride (.10 M), is added tetrazole (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N,N-diisopropyl-chlorophosphoramidite ( 1.5 equiv.) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 198~, 12, 4539.
After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( lX) and is then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 181(66% yield). scheme 21 Preparation of 182 (R group = OTES, NPhth or NHAc) To a solution of triol 178 (.0 equiv.) in CH2Cl2 (.5 M) at 0 ~C, was SUBSTITUTE SHEET (RULE 26) W096~7379 r~l/

added triethylamine (1.2 equiv.), 4-DMAP (.10 equiv.) and then TOSCl ( 1.1 equiv.). The reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (lX 5 mL), back-extracted (2X), recombined, dried (MgSO4) and evaporated. The compound is purified by flash column chromatography and then a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (2.2 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (2.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords the protected tosylate/acid 182.

Preparation of 183 (1~ group = OTES. NPhth or NHAc) To a solution of triol 182 (.0 equiv.) in CH2C12 (.5 M) at 0 ~C, is added sodium-azide ( 1.2 equiv.) from Aldrich chemical company at 0 ~C. The reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (lX 5 mL), back-extracted (2X), recombined, dried (MgSO4) and evaporated.
The compound is purified by flash column chromatography and SUBSTITUTE SHEET (RULE 26) WO96~?379 ~ U~C~o3227 _99_ affords compound 183.

Preparation of 18~ (R group = OTES, NPhth or NHAc) A solution of 183 (1.0 equivalents) in ethanol (.01 M total) at 25 ~C is exposed to 10% Pd(OH)2-C (0.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere.
The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 185 scheme 21.

Preparation vf 191 H O ~ ~~
~ 'OH
OH

A solution of starting material 19 0 as disclosed by Schmidt, R. R. et al. (Liebigs Ann. Chem. 1 9 8 7, 825), ( 1.0 equivalents) is dissolved in a mixture of ethanol-tetrahydrofuran-acetic acid (2:1:1), (.01 M total) at 25 ~C. The " mixture is next exposed to 10% Pd/C ( 1.0 equivalents) and is SUBSTITUTE SHEET tRuLE 26) W096127379 ~ 96/03227 then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 100% ethyl acetate in petroleum ether) affords compound 191. scheme 22 step a SUE~STITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096~7379 r~l~U~36,'~3~27 Preparation of 192 r TsO ~O
TES~ ~ OTES
OTES

To a solution of 191 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added tosylchloride ( 1.2 equivalents) at 0 ~C. Subsequent addition of triethylamine ( 1.5 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated to afford the crude tosylate. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 192. scheme 22 step b SUI~STITUTE SHEET (RULE 26) CA 022l4789 l997-09-0=, W096~ m 9 ~ ,''03227 Preparation of 193 N3--~~

TESC~ ~ OTES
OTES

To a solution of 192 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added sodium azide from Aldrich chemical company ( 1.2 equivalents) at O ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with ammonium chloride (2X), brine ( lX) and then M g S 0 4 . The solution is then concentrated and purification by flash column chromatography affords compound 193. scheme 22 step c Preparation of 194 N3--~o~ ~CO2H

TESb~ ~ OTES
OTES

To solution of 193 in CCl4 (.33 M), CH3CN (.33 M) and water (.22 M) at O ~C is added RuCl3 (.03 equiv.) and NaI04 (4.0 equiv.) and the muddy black mixture is allowed to stir for 1.5 h.
The mixture is then diluted with ether (25 mL), washed with SU B STITUTE S H EET (RU LE 26) WO 9612m9 ~ 03227 water (2X 5 .0 mL) and brine ( lX 5 mL). The aqueous layer is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 194. scheme 22 step d Preparation of 196 FMOCNH ~O~ "CO2H

TESC~ 'jJ OTES
OTES

A solution of 194 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 ~C. The mixture is next exposed to 10% Pd/C
(.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine ( lX) and dried (MgSO4 ) and concentrated. Next, to a solution of crude amine (1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-C1, 1.2 equivalents) is followed by stirring for 2 hours SIJBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W 096/27379 PCTnUS96/03227 and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried ( M g S 0 4 ) and concentrated. Purification by flash column chromatography affords compound 196. scheme 22 steps e-f s SU~STITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096127379 ~ 96J03227 Preparation of 197 DMTo~O~.~

TESo'~f OTES
OTES

To Tetrol 191 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgSO4 and concentrated.
Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 197. scheme 22 step g S~J~STITUTE SHEET (RULE 26) CA 022l4789 l997-09-0=.
W096127379 ~ U~ 3227 Preparation of 198 DMTO ~o~ c02CH3 TESC~ ~J OTES
OTES

s To solution of 197 in CCl4 (.33 M), CH3CN (.33 M) and water (.22 M) at 0 ~C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for l.5 h.
The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine ( lX 5 mL). The crude is then resuspended in a mixture of methylene chloride/water ( l :1, . l M total) and diazomethane ( l .2 equivalents) is gradually dropped into the flask via an addition funnel at the rate of l drop/10 seconds. After complete addition the mixture is diluted lS with ether, washed with brine (2X) and the aqueous layer is back extracted (2X) recombined, and the organic layer is then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 198. scheme 22 step h SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096~7379 ~-~JU~ D3227 Preparation of 200 D MTO ~O~ ~C H20 H

TESC~ ~J OTES
OTES

To a solution of 198 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added a l.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X), brine (lX) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 200. scheme 22 step i lS Preparation of 201 DMTO--~O~ N3 TES~ ~ OTES
OTES

A solution of 2 0 0 ( 1.0 equivalents) in tetrahydrofuran (. l 8 M) is treated with DPPA (diphenylphosphorylazide, 2.0 SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096127379 ~ 03227 equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD
(diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is heated to 80 ~C for 3 hours and then diluted with ether (2X) and washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgSO4 and evaporated. Purification by flash column chromatography affords compound 201. scheme 22 step j Preparaffon of 202 DMTO--~O~ N H FMOC

TESC~ ~IJ OTES
OTES

A solution of 201 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 ~C. The mixture is next exposed to 10% Pd/C
(.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atrnosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine ( lX) and dried (MgSO4 ) and concentrated. Next, to a solution of crude amine (1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-CI, 1.2 equivalents) is followed by stirring for 2 hours SUBSTITUTE SHEET (RULE 26) W096~1379 r~Ju~5~D3227 and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( 1 X) and then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography affords compound 202. scheme 22 step e s Preparation of 204 HO2C----~O~ NHFMOC

TES~ ~ OTES
OTES

To asolution of 202 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added 10% HCOOH from Aldrich chemical company ( 1.2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium bicarbonate (2X), brine ( lX) and then MgSO4. The solution is then resuspended in t-BuOH (.10 M) and pH 7 buffer (.10 M) and is then exposed to KMnO4 ( 1.2 equivalents) for 2 hours at 0 ~C. The reaction mixture is next washed with sodium bicarbonate (2X), brine (lX) and then MgSO4. The organic layer is then concentrated and purified by flash column chromatography to afford compound 204. scheme 22 step k 5UBSTlTUTE SHEET (RULE 25~

CA 02214789 1997-09-0~
W096127379 ~ 7 Preparation of 206 DMTO--~O~--NO2 TESO~ OTES
OTES

To Tetrol 205 (1.0 equivalents) (as disclosed by Petrus, L.
et ahChem. zvesti. 1982, 36, 103) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgS04 and concentrated.
Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 206. scheme 23 step a SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~ -W~9612~379 ~JU~,~103227 _111_ Preparation of 207 DMTO~O~--NHFMOC
TESO~J OTES
OTES

Toa solution of 206 (1.0 equivalents) in diethylether (.08 M), is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 ~C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine (lX) and is then dried (MgS04) and concentrated. The crude mixture is resuspended in methylene chloride (.10 Molar) and to it is added sodium bicarbonate (2.0 equivalents) at O ~C.
Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 207. scheIne 23 step b 5U13STITUTE S~tE~T tRULE 26) CA 022l4789 l997-09-05 W096127379 P~ ,''03227 Preparation of 208 HO--I'O~--NHFMOC
TESO~ OTES
OTES

To a solution of 2 0 7 ( 1.0 equivalents) in methylene chloride (.10 Molar) is addedlO% HCOOH (1.1 equivalents). The reaction is stirred at 0 ~C for 2 minutes and is then diluted with ether and washed with sodium bicarbonate (2X), brine ( 1 X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 208. scheme 23 step c Preparation of 209 HOOC~O~ NHFMOC

TESO~ 'OTES
OTES

To solution of 208 in CC14 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 ~C is added RuC13 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine ( lX 5 mL). The aqueous layer SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W096~73~9 ~ 6103227 is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 209. schelme 23 step d Preparation of 210 TOSO--~O~--NO2 TESO~ OTES
OTES

To a solution of 205 (1.0 equivalents) in methylene chloride (.10 Molar) is added tosylchloride ( 1.2 equivalents) at 0 ~C. Subsequent addition of triethylamine ( 1.5 equivalents) is followed by stirrin~; for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated to afford the crude tosylate. Next a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with SUI~STITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W096~7379 ~ ''03227 ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 210. scheme 23 step e Preparation of 211 N3~ ~ NO2 TESO~ OTES
OTES

To a solution of 210 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added sodium azide from Aldrich chemical company ( 1.2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with ammonium chloride (2X), brine (lX) and then M g S O 4 . The solution is then concentrated and purification by flash column chromatography affords compound 211. sche~ne 23 step f SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W 096127379 l~ '3~103227 Preparation of 212 N3,~0rCO2H
TESO~ OTES
OTES

To solution of 211 in CCl4 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 ~C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5 .0 mL) and brine ( 1 X S mL) . The aqueous layer is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 212. scheme 23 step g Preparation of 213 H2N ~O~I~C02H

TESO~ OTES
OTES

A solution of 212 (l.0 equivalents) in ethanol (.01 M total) at 25 ~C is exposed to 10% Pd/C (0. l equivalents) and is then SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
WO961Zn79 ~l/U~,''03227 subsequently capped with a hydrogen balloon at 1 atmosphere.
The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (lX) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 213. scheme 23 step h Preparation of 214 FMOCH N ~O~CO2H
TESO~ 'OTES
OTES

Compound 213 is suspended in methylene chloride (.10 Molar) and to it is added sodium bicarbonate (2.0 equivalents) at 0 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( l X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 214. scheme 23 step i SUBSTITUTE S~EET (RULE 26) CA 022l4789 l997-09-0~
W096~7379 ~ ~9~D3227 Preparation of 215 P IVO--~O~--NO2 TESO~ OTES
OTES

To a solutionof 205 ( 1.0 equivalents) in pyridine (.10 Molar), is added trimethylacetyl chloride (pivaloyl chloride) (2.5 e~uivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgSO4 and concentrated. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 215. scheme 23 step j SlJBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W 096~7379 ~ '3~103227 Preparation of 216 PlVO--~O~C02M e TESO~ OTES
OTES

To solution of 215 in CCl4 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 ~C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5 .0 mL) and brine ( 1 X 5 mL) . The aqueous layer is back extracted (2X), recombined, and the organic layer is then dried MgSO4 and evaporated. The crude is then resuspended in a mixture of methylene chloride/water ( 1: l, .1 M total) and diazomethane ( 1.2 equivalents) is gradually dropped into the flask via an addition funnel at the rate of l drop/ l 0 seconds.
After complete addition the mixture is diluted with ether, washed with brine (2X) and the aqueous layer is back extracted (2X) recombined, and the organic layer is then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 216. scheme 23 step k SUBSTITUTE StlEET (RULE 26) W096~7379 ~-lJU'~ 3227 _1 19--Preparation of 217 .

Plvo - ~o~cH2oH
TESO~J OTES
OTES

To a solution of 216 ( 1.0 equivalents) in methylene chloride (. l 0 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( l .2 equivalents) at 0 ~C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X), brine (lX) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 217. scheme 23 step l Preparation of 218 PIVO--~CH20DMT
TESO OTES
OTES

To 217 (l.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) ( l .1 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with SUBSTITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
W096~ m g ~ 6tO3227 diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgSO4 and concentrated.
Purification by flash column chromatography affords compound 218. scheme 23 step m Preparation of 220 Ho - l~o~l - cH2oDMT
TESO~J 'OTES
OTES

To a solution of 218 (1.0 equivalents) in diethylether (.08 M), is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 ~C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine ( 1 X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 220. scheme 23 step n SUBSTITUTE S~l EET (RU LE 26) WO 96t2mg ~I_lJ~Y6J1~3227 Preparation of 221 D MT~X~~ N ~2 TESO 'OTES
OTES

To Tetrol 20~ (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 ~C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (lX), dried over MgSO4 and concentrated. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 ~C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 221. scheme 23 step a SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096127379 ~-l/U~ 03227 Preparation of 222 DMTO~O~C02H

TESO~J OTES
OTES

To solution of 221 in CC14 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 ~C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine (lX 5 mL). The aqueous layer is back extracted (2X)~ recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 222. scheme 23 step o.

Preparation of 224 TESOX~'OTES
OTES

To a solution of 222 ( 1.0 equivalents) in diethylether (.08 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
wo96~73n ~ U~6103227 M), is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 ~C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine ( 1 X) and Sis then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 224. scheme 23 step p Preparation of 216 To a stirred solution of the acid 214 ( 1.0 equivalents) in dimethylformamide (.10Molar) at 25 ~C, is added 1-hydroxybenzotriazole(HOBT; 1.1 equivalents). Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 1 hour in the presence of an appropriately 15substituted solid support (N-(2-Aminoethyl)-3-amino-propyl glass; aminopolystyrene resin; aminopropyl glass; isothiocyanato glass, all with or without a linker extending from the amino group on the support etc. from Sigma Company). The mixture is then diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated.

Preparation of 226; 228; 230 or 232 To a stirred solution of the acid 214; 62; 215 or 62 ( 1 . 0 SUI~STITUTE SHEET (RULE 26) CA 02214789 1997-09-0~
WO9612?379 PCTrUS96103227 equivalents) and the amine 216; 226; 228 or 230 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 ~C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether~ filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over M g S 0 4 and then concentrated. Purification by flash column chromatography and then reexposure of the intermediate compound (1.0 equivalents) in dimethyl-formamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CuSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated.
Purification by flash column chromatography affords compound 226; 228; 230 or 232, respectively. scheme 24 Preparation of 234 To a stirred solution of 232 ( 1.0 equivalents) in acetonitrile (.50 Molar) is added an HF pyridine solution (.50 M) from Aldrich chemical company. The reaction is allowed to stir for five hours and is then condensed. The crude 234 oligomer is then resuspended in p-dioxane (.50 Molar) to which is added a 3.0 Molar solution of NaOH (3.0 equivalents). The reaction is stirred SUBSTITUTE SHEET (RUEE 26) WO 96127'379 ~ U~56~03227 for 1 hour at SO ~C and is then quenched with aqueous NH4C l (2X) and subsequently Iyophilized. Purification by HPLC
chromatography affords compound 234. scheme 24 SUBSTITUTE SHEET (RUI_E 26) CA 02214789 1997-09-0~
WO 96127379 ~ l/U~ '??7 Preparation of Peptoid Combinatorial libraries (Scheme 500) A depiction of the generation of a combinatorial library for oligopeptoid compounds is shown in scheme 500. The example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itsel~ to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for peptoid synthesis. The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. ~ll supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).

SUI~STITUTE SHEET (RULE 26) wa 96~7379 ~-11u-_~D3227 cq ~:~~ 2i~ ~--~~ '~ \

~ ~o c~~~~ 1' 0~0 ~Z 0 , ~ ~O

O ~ ", ~ I ~

~a O ~ Q 5 ~ ~ ~

~ ~ ~ ~' ~

O ~, ~L I ~

~Q ~

O ~ I O

SUBSmU~E SHEET (RULE 26) CA 022l4789 l997-09-05 W096127379 ~ 96103227 o ~ Ga I ~

o~@ ~ d~ ' ~ ~

O C/~ o ~0 X
C~-O~ ~C~"_ E ~ E

E_ ~" 'Z ~ G;;l I ~

~ Z

C~-~ C~ ~ I . X x UJ n O C~ o C~O ~ ' X X 1~
_I O ~ ~ t7 - ' -- 0 LL

SUBSTITUTE SHEET (RULE 26) .

WO 96/27379 ~ iu~56JD3227 C ~
X X 1l \ Z
T O ~ U~ O=~U~

O~u z ~ I c Q \ ~ ~~

~ b ", ~ r n ~ ~ ~
I . X

~ ~ o C~ \ N
x m a ~ Q . a ~
rD ~ X ' N~
Q ~ _ c~ _ O L' D C~l I 11, C~ o ~ ---- u~
X
_~ aJ /
C., "_ ~I ~ /
l-- O ~
O ~ XX m m ~,. m Z

O ~ c ~n C~ . X
N X

SUBSmUTE SHEET (RULE 26) W 096127379 ~ 6/03227 Z Z

~ ~Z @ ~ . 8 ~

~~ ~ ~

o ~- 9 ~ ~ o ~1 ~o ~ 0 ~ ~~

d ~ o ~~ O ~ ~ o ~

~z ~ /
~0 ~ o ~u~ E ~ ~ O~O m a ~ o@ ~
o~ o ~0 I UJ

SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096~7379 ~-1J~3~D3227 ~ llJ ~ ~ ~ X _ I
O O 0 t ~~ t ~ ~ ~l \
cn O O ~ O. ~ ~ ~ o ~ ~L~ t Z i~/ ~ ~ ~ G~ I t ID Z ~0~-o ~ ~ = Z 0 C~ ~ ~ ,c ~
UJ~ N ~ C E tn ~~C~

,N5 1-- N N

~ O ~ ~, . ~q /

o-~L cn o ~ ~ ~ g N X I /
0~0 O ~J~

SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W 096/27379 l~l/u~,G1~227 N
N N N j~ I CL ~~

~W ~0 ~ ~ ~ X ~ ~

o d oO~ l 0 ~~~

~ ~ e~ o ~ ~ IL 0~
~ ~ Q E ~
UJ - 'S ~ _ _ \

~0 ~ ~ID XX ~, _- ~ ~ o~ Z O

' z 0 c~ o m t , x x a~ E
- c' o x u c ~0 _ _, ~ x, - ",'~ a~
DE r ~ Q G
~Z0~ - .~ t x xN 11 /

Q ~ ~ = ~ E-~: m O ~ --~ ~

~ Z 0 C) ~ E~ . X x ~
~--O ~ ~
~ 0 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W096~7379 ~ 6'C~?7 Preparation of Nucleotoid ~ombinatorial libraries (Scheme 550) A depiction of the generation of a combinatorial library for S oligonucleotoid compounds is shown in scheme 550. The example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itself to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for carbonucleotoid synthesis. The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support;
amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol~ bromide etc.).
Preparation of compound 2000 . To a solution of 7 6 ( 1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde 1.1 equiv), and the solution was exposed to ZnCl ( 1.1 equiv) at 25 ~C and allowed to stir for 2.5 hour. The solution is then SUBSTITUTE SHEET tRULE 26) CA 022l4789 l997-09-05 W096/2 m 9 ~ ,Cl03227 diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine ( 1 X) and then dried over MgS04. The compound is purified by flash column chromatography to yield the desired benzylidene.

SUBSTITUTE SHEET tRULE 26) -W096~7379 ~-1lu~Y6JD3227 1 Synthesis of a C-2 differentiated sugar HO~O C02Et 1. PhCHO, ZnCI, CH2C~Orco2Et HO J~OH ~ Ph O'~'OH
HO 2. Bu2SnO, MeOH OBn 76 then BnBr, CsF, DMF 2C00 3. NaCNBH3 (~ eq), DMF
(3 A sieves, TFA 10 eq) BnO~O o 4. PhCHO,ZnCI, CH2C12 BnO~l~O~I~oH
HO'~O Ph OBn 5. BnBr, NaH, THF

BnO~O~I - o 6. NBS, BaCO3 BnO~Ol~OH

OBn CCI4-C2H2Cl4~ H2O OBn 2030 7 o~CN

Cl' Ni(Pr)2 CH2CI2, (iPr)2EtN
o~CN
BnO~O~~O~ Ni(Pr)2 Bz = Benzoate BnO ~'OBz Bn = Benzyl ether OBn 20~0 Scheme 2000a SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 WO 96127379 r~~ 9~03227 2.Connection of the C-2 differentiated sugar to a solid support BnO ~ OH 1. NaH, THF BnO~o ~N~ Solid Support ¦
Bn~'OBz BnO 'OH ~
OBn 2. then add to OBn 2040 Br ~N~ Solid Support ¦ 2060 3. 1% NaOH, MeOH

Scheme 2000b SUBSTITUTE SHEET (RlJLE 26) CA 022l4789 l997-09-0~
W096/Z7379 ~llU~,~6103227 " The benzylidene is then azeotroped with benzene (2X 100 mL)and then dried overnight under vacuum over P2 O 5 . A mixture of benzylidene, dibutyl tin oxide ( 1.2 equiv.) and dry methanol (.25 M) are heated at reflux for 4 h until the solution became clear and homogeneous. (An automatic stirring apparatus may be necessary.) The solvent is next removed in vacuo to give a foamy white tin complex which was then azeotroped with benzene (2X) and dried (2 h to overnight) under vacuum over P2Os. Next, anhydrous DMF (.2M) is added to redissolve the tin complex and then CsF ( 1.2 equiv.) and finally Benzyl bromide (1.5 equiv.) are added and then heated (40 ~C) overnight. The clear solution is partially distilled under vacuum, (3.3 mm Hg, 75-100 ~C) to obtain 1/5 the original volume of solvent.
Reaction mixture was then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) to remove cesium salts. Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2000. For related chemistry see Nagashima, N.; Ohno, M. Chemistry Letters, Chem. Soc. of Japan 1987, 141.

S~JBSTITUTE SH EET (RULE 26) CA 022l4789 l997-09-0~
W 096~7379 ~ 61o3227 Preparation of compound 2010.
Procedure adopted from Johansson R.; Samuelsson; B. J. Chem.
Soc., Chem. Commun., 19 8 4, 201. To a solution of the benzylidene acetal ( 1 equiv) and sodium cyanoborohydride (5 equiv.) in DMF (.125 M) containing powedered 3 angtrsom molecular sieves is added trifluoroacetic acid ( 10 equiv) and the reaction is allowed to stir at 0 ~C until no starting material remains. Reaction mixture is then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2010.

Preparation of compound 2020.
To a solution of 2010 (1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCl ( 1.1 equiv) at 25 ~C and allowed to stir for 2.5 hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (lX) and then dried over MgSO4. The compound is purified by flash column chromatography to yield the desired benzylidene 2020.

SlJBSTITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
wa s6t2ms ~-h~u:3~6Jr~7 Preparation of compound ~030.
To a solution of alcohol 2020 (22.0 g, .1068 mol, 1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction iss cooled to 0 ~C and treated with benzyl bromide ( 1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (lX), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2030.

Preparation of compound 2040.
Procedure as adopted from Hanessian S.; Organic Synt~eses 19 8 7, 243. To a suspension containing 1.0 equivalent of benzylidene 2 0 3 0 in one molar carbon tetrachloride and 1,1 ,2,2-tetrachloroethane ( 1.5 equivalent) is added 1.2 equivalents of N-bromosuccinimide and 0.5 equivalents of barium carbonate. The resulting suspension is heated at the reflux temperature of the mixture with mechanical stirring for a period of 2.5 hour and filtered while hot. The solution is washed with water (3X), then dried over anhydrous sodium sulfate and evaporated. Purification by flash column chromatography yields SUBSTITUTE SH EET tRULE 26) CA 022l4789 l997-09-05 W096~7379 r~l/U~96/03227 tribenzyl ether 2040.

Preparation of compound 2050.
To a solution of 2040 (1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res.
1984, 12, 4539. After 15 minutes the reaction is SUBSTITUTE StlEET (RULE 26) CA 022l4789 l997-09-05 W096~7379 ~ /03227 -14t-Synthesis of a C1-C2-Phosophodiester oligomer using a solid support o~CN
Bno~ol - o~pNi(pr)2 BnO~O~--o~~lrN--¦ Solid Sup )ort BnO'~'OBz BnO

1. tetrazole, CH3CN
2. MCPBA, CH2Clz ~ 3. 1% NaOH, MeOH, 20OC

BnO~o ~N--¦ Solid Sup )ort BnO ~ O
o p_o~CN
Bno~~l~d BnO' i 'OBn OH

1. 2050. then tetrazole, CH3CN
2. MCPBA, CH2C12 ~ 3. 1% NaOH, MeOH, 20~C

BnO~O~_o ~N--¦ Solid Sup ~ort BnO' ~3 ~
O FI_o~CN
Bno~~l~d BnO'~3 BnO
O-F ~_o~CN
Bno~~~~d ,~BnO'~'OBn OH
I TERA TE
.. .

Scheme 2001 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W 096~7379 ~ U~ 3227 brine (lX) and is then dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 2050 (as shown in scheme 2000).
s Preparation of compound 2060 To a solution of alcohol 2040 (1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 ~C
and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached ( 1.0 equiv . ) and stirred for 2 hours. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 ~C and the support was washed with ethyl acetate, 1% NaOH in methanol (2X) to remove the benzoate and finally brine ( 1 X) to give 2 0 6 0 . The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amzno group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).

SUBSTITUTE SHEET (RULE 26 CA 022l4789 l997-09-05 W096~7379 ~ 0~ 7 1. Synthesis of a C-3 ~lir~ , ILidLed sugar 1~

HO~CO2Et 1. PhCHO. ZnCI. CH2C12 ~'oH

HO 2 NaCNBH3(5 eq), DMF 2070 76 (3 A sieves, TFA 10 eq) 3. TBDPSCI, Et3N, CH2C12 4. Bu2SnO, MeOH, BzBr, CsF, DMF
'1 BnO~OH 5. BnBr, NaH. THF BnO~OTBDPS

BzO 6. TBAF. THF OBz 2~90 2080 o~CN
7. ,p .
Cl N'(Pr)2 y CH2CI2, (iPr)2EtN

o~CN

BnO~rOrO~ N (Pr)2 Bz = benzoate BnO ~~'OBn Bn = benzyl BzO

Scheme 2002a SUBSTITUTE SHEET (RULE 26) _ _ , WO 96127379 ~ 961Q3227 Z.Connection of the C-3 .li~el ~l ILid~d sugar to a solid support BnO~OH 1. NaH, THF BnO~~o ~N--l SoGd Suppor BzO 2. then add to HO
2090 Br~,N--l Solid Suppo~ 2110 o 3. 1% NaOH, MeOH

S~heme 2002b SUBSTITUTE 5HEET (RUL~ 26) CA 022l4789 l997-09-0~
W096~7379 ~ 03227 Preparatfon of compound 2070 " To a solution of 76 (1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCl (1.1 equiv) at 25 ~C and allowed to stir for 2.5 hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (lX) and then dried over MgSO4. The compound is purified by flash column chromatography to yield the desired benzylidene.
Procedure adopted from Johansson R.; Samuelsson; B. J. Chem.
Soc., Chem. Commun., 19 8 4, 201. To a solution of the benzylidene acetal ( 1 equiv) and sodium cyanoborohydride (5 equiv . ) in DMF ( .125 M) containin~, powedered 3 angtrsom molecular sieves is added trifluoroacetic acid ( 10 equiv) and the reaction is allowed to stir at 0 ~C until no starting material remains. Reaction mixture is then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2070.

Preparation of compound 2080 To a solution of 2070 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added triethylamine ( 1.1 equivalents) at 0 ~C.

SU~STITUTE SHEET (RULE 26) , CA 022l4789 l997-09-0~
W 096r27379 PCTÇUS96/03227 Subsequent addition of tertbutyldiphenylsilylchloride ( 1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords the TBDPS ether which is subsequently carried on as follows:
The TBDPS ether is then azeotroped with benzene (2X 100 mL) and then dried overnight under vacuum over P2 O 5 . A mixture of benzylidene, dibutyl tin oxide ( 1.2 equiv.) and dry methanol (.25 M) are heated at reflux for 4 h until the solution became clear and homogeneous. (An automatic stirring apparatus may be necessary.) The solvent is next removed in vacuo to give a foamy white tin complex which was then azeotroped with benzene (2X) and dried (2 h to overnight) under vacuum over P2O5. Next, anhydrous DMF (.2M) is added to redissolve the tin complex and then CsF ( 1.2 equiv.) and finally Benzoyl bromide for the benzoate formation, ( 1.5 equiv.) are added and then heated (40 ~C) overnight. The clear solution is partially distilled under vacuum, (3.3 mm Hg, 75-100 ~C) to obtain 1/5 the original volume of solvent. Reaction mixture was then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) to remove cesium salts. Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W 096~7379 r~-l/U~6/03227 organic layer which was then dried over MgSO4 and evaporated.
Purification by flash column chromatography yields the desired benzyl ether 2080. For related chemistry see Nagashima, N.;
Ohno, M. Chemistry l etters, Chem. Soc. of Japan 1987, 141.
s lPreparation of compound 2090 To a solution of alcohol 2080 (1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over severaI portions. The reaction mixture is warmed to room temperature and stirred 1 h. Next, the reaction is cooled to 0 ~C
and treated with benzyl bromide ( 1.0 equiv.) and stirred for 1.5 h. The compound is then treated with tetrabutylammonium fluoride (2.0 equivalents) and allowed to stir for an additional 2 hours. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine ( lX), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2090.

Preparation of compound 2100 To a solution of 2090 (1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as SUE~STITUTE StlEET (RULE 26) CA 022l4789 l997-09-05 W096127379 PCT/u~ C~7 prepared from the procedures of Sinha et al. Nucl. Acids Res.
1984, 12, 4539. After 15 minutes the reaction is complete and .
is next diluted with ether and next washed with brine (lX) and is then dried (MgSO4) and concentrated. Purification by flash S column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 2100 (as shown in scheme 2002).

SUE~STITUTE SHE~:T (RUL~ 26) W096127379 ~ /U~961o3227 Synthesis of a C1-C3-Phosophodiester oligomer using a solid support o~CN
BnO~--o~PNi(Pr)2BnO'~-O~ o~~N--¦ Solid Supp )rt BnO 'OBnBnO'~'OBn ~
J 'BZO OH

1. tetrazole, CH3CN
2. MCPBA. CH2CI2 ~ 3. 1% NaOH, MeOH, 20~C

BnO~O~ - O - ~N-- Solid Supp )rt BnO'~'OBn ~
O=P--o~CN
BnO~O~
BnO'~f 'OBn OH

1. 2100, then tetrazole, CH3CN
2. MCPBA, CH2C12 ~ 3. 1% NaOH, MeOH, 20~C

BnO~~o ~N--¦ Solid Supp ~rt o O=P--o~CN
Bno~~~d BnO~ OBn O=P--o~CN
Bno~~l~d BnO'~'OBn OH /TERATE

Scheme 2003 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W096/27379 ~-1J~3G/03227 Preparation of compound 2110 To a solution of alcohol 2090 (1.0 equiv.) in THF ~0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred 1 h. Next, the reaction is cooled to 0 ~C
and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached ( 1.0 equiv.) and stirred for 2 hours. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 ~C and the support was washed with ethyl acetate, 1% NaOH in methanol (2X) to remove the benzoate and finally brine ( 1 X) to give 2110. The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support f eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
Preparation of compound 2120 To a solution of 7 6 ( 1 .0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCl ( 1.1 equiv) at 25 ~C and allowed to stir for 2.5 SUBSTITUTE SHEET (RULF 26) CA 022l4789 l997-09-05 W096~7379 ~-lIV~96/03227 hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (lX) and then dried over MgS04. The compound is J purified by flash column chromatography to yield the desired SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096/27379 PCT/u~ tt7 1. Synthesis of a C-4 differentiated sugar HO~CO2Et 1. PhCHO, ZnCI, CH2CI2 phl~'OBn 7H6O 2. BnBr, NaH. CH2C12 2120 3. NaCNBH3 (5 eq), DMF
(3 A sieves, TFA 10 eq) BnO~OTBDPS 4 TBDPSCI, Et3N, CH2CI2 gno~OTBDPS
BzO 'OBn HO 'OBn OBn 5. BzBr, NaH, THF OBn 6. TBAF, THF

BnO~OH 7- Cl'P ~ (P" BnO~ o~CN
OBn CH2CI~ Pr)2EtN OBn Bn = benzyl Scheme 2004a SU~STITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 wog6n7379 ~ U~C~3227 '~ 2.Connection of the C-4 differentiated sugar to a solid support BnO~rO~~OH 1. NaH, THF BnO~O~~o~~N--l Solid Supp )rt BzO y' OBn HO I 'OBn ~
OBn 2. then ~ 1 lo OBn 2150 Br ~N-- Solid Supp )rt 2170 3.1% NaOH, MeOH

Scheme 2004b SUSSTITUTE SHEET (RUL~ 26) CA 022l4789 l997-09-0~
W096/2n79 ~-l/U~/03227 benzylidene and carried on as follows:
To a solution of benzylidene (1.0 equiv.) in ~HF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 ~C
and treated with benzyl bromide ( 1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine ( lX), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2120.

Preparation of compound 2130 Procedure adopted from Johansson R., Samuelsson; B. J. Chem.
Soc., Chem. Commun., 19 8 4, 201. To a solution of the benzylidene acetal 2120 ( 1 equiv) and sodium cyanoborohydride (5 equiv.) in DMF (.125 M) containing powedered 3 angtrsom molecular sieves is added trifluoroacetic acid ( 10 equiv) and the reaction is allowed to stir at 0 ~C until no starting material remains. Reaction mixture is then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by SUBSTITUTE SHEET (RULE 26~

CA 022l4789 l997-09-0~
W096~7379 ~ Y6103227 flash column chromatography yields the desired benzyl ether 2130.

Preparation of compound 2140 To a solution of 2130 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added triethylamine ( 1.1 equivalents) at 0 ~C.
Subsequent addition of tertbutyldiphenylsilylchloride ( 1. l equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (lX) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords the TBDPS ether which is subsequently carried on as follows:
To a solution of TBDPS ether (1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred 1 h. Next, the reaction is cooled to 0 ~C
and treated with benzoyl bromide to afford benzoate formation ( 1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (lX), dried over M g S O 4 and evaporated. Purification by flash column chromato~raphy yields tribenzyl ether 2140.

,.

SUBSTiTUTE StlEET ~RULE 26) CA 022l4789 l997-09-0~
W096/27379 r~l/U~6/03227 Preparation of compound 2150 The compound 2140 is then treated with tetrabutylammonium fluoride (2.0 equivalents) in THF (.1 Molar) and allowed to stir for an additional 2 hours at 25 ~C. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (lX), dried over M g S O 4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2150.

Preparation of compound 2160 To a solution of 215 0 ( 1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 ~C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res.
1984, 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( l X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 30~o ethyl acetate in petroleum ether) affords compound 2160 (as shown in scheme 2004).

SUBSTITUTE SHEET (RULE 26) -CA 022l4789 1997-09-OF.
WO 96t2mg PCr/~S96~03227 Preparation of compound 2170 To a solution of alcohol 2150 (1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 ~C
and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached ( 1.0 equiv.) SU B STITI JTE SH EET (RU LE 26) CA 022l4789 l997-09-05 WO9612 m 9 ~tLllU~,''03n7 Synthesis of a C1-C4-Phosophodiester oligomer using a solid support o~CN
BnO~O~o' Ni(Pr)2BnO~~O~l o ~N--~ Solid Supp ~rt BzO' ~ 'OBnHO'~f'OBn ~
BnO BnO

I

1. tetrazole, CH3CN
2. MCPBA. CH2CI2 ~, 3. 1% NaOH, MeOH, 20~C

BnO ~O ~fN--¦ Solid Supp 7rt ~' 'OBn O
¦ BnO CN
BnO~O~I J
HO'~'OBn BnO

1. 2160, then tetrazole, CH3CN
2. MCPBA. CH2C12 ~, 3.1% NaOH, MeOH, 20~C

BnO~o~~N--¦ Solid Supp )rt ~' 'OBn ~
¦ BnO CN
BnO~O~--J
~;)'~'OBn I BnO
O=P--o~CN
BnO~O~--J
HO'~'OBn OBn tTERATE

Scheme 2005 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096~7379 ~ 7~61o3227 and stirred for 2 hours. A saturated solution of ammonium Y chloride (50 mL) is added dropwise to quench the reaction mixture at 0 ~C and the support was washed with ethyl acetate, 1% NaOH in methanol (2X) to remove the benzoate and finally brine (lX) to give 2170. The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as pllrchased from Sigma company. All supports may be with or without a linker extending from the amino group on the 0 support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide elc.).

Prepartion of compound 3010 Procedure as described in Methods in Carbohydrate chemistry, Whistler, R., II, 1963, p. 327. A mixture of 80 g anhydrous D-glucosamine hydrochloride or D-galactosamine hydrochloride from Aldrich chemical company, in 200 mL. methanol and 20g Dowex 50 (H+) acidic resin, is stirred at the boiling point in a round bottom flask. After 24-hr. reaction time, the resin is removed by filtration and ished three times with 20 ml. of methanol. The filrate and washings are combined and concentrated to about 125 ml by rotovap. The concentrate is allowed to cool to room temperature and the product crystallizes overnight and carried on as follows:

SUBSTITUTE Sl IEET (RULE 26) CA 022l4789 l997-09-05 W096127379 PCTnUS96/03227 The methyl glycoside is dissolved in chloroform (.S M) and to it, is added phthalic anhydride ( 1.5 equiv.) and the reaction mixture is allowed to reflux at 70 ~C for 4 h. The product SUBSTITUTE SHEET (RUI_E 26) CA 02214789 l997-09-05 W096~7379 PCTnUS96103227 1. C-2 differentiated amine derivative Ho'~,~r~H 1. MeOH, H+ HO~OrOMe 3- BnBr, NaH BnO~OrCO2Et HO ~ Y HO 4. TMSCN, SnC4 BnO

Glucose amine hydrochloride 3010 5. EtOH, H2SO4 (cat) 6. FMOC-CI, K2CO3, THF, H2O, 0 ~C.
~ 7. NaOEt, MeOH
BnO~O C02H
BnO'~XNHFMOC
BnO 3030 2. Connection of a C-2 dirr~ liaL~ amine sugar to a solid support BnO~O CO2H 1. DCC, HOBT, Et3N, DMF ~
r ~ BnO'~O~o_ Solid Su~ ~ort BnO' ~ NHFMOC
BnO I BnO' '~'NHz 3030 HO--lSolid Supp )rtBn3oo4o 2. Piperidine, DMF

Scheme 3000 SUE~STITUTE SH EET (RULE 26) CA 02214789 1997-09-0~
W 096~7379 ~ 1U~g6/03227 3010 is then crystallized and carried onto the next step.

Preparation of compound 3020 To a solution of alcohol 3010 (1.0 equiv.) in THF (0.5 M) at 0 ~C, is added NaH (3.3equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 ~C
and treated with benzyl bromide (3.3 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 ~C and the mixture was diluted with ethyl acetate, washed with water (2X), brine ( lX), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether and is carried on as follows:
To a solution of tribenzyl ether in nitromethane is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the rem~ining oil is resuspended in dichloromethane and washed with sodium acetate solution (lX), water (lX), brine (lX) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol is next dissolved in ethanol (0.15 M) and then concentrated H2SO4 (0.01 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W0961~7379 ~ C~03227 equivalents-catalytic) is added. The reaction mixture is heated to 85 ~C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography affords compound 3020 scheme 3000.

Prepartion of compound 3030 To a solution of 3020 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added potassium carbonate (2.0 equivalents) at 0 ~C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine ( lX) and then dried ( M g S 0 4 ) and concentrated. Purification by flash column chromatography affords product which is carried on as follows:
To a solution of ester in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column chromatography affords compound 3030 scheme 3000.
Preparation of compound 3040 To a stirred solution of the acid 3030 ( 1.0 equivalents) and the ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 ~C~ is added l-hydroxybenzotriazole (HOBT; 1.1 equivalents). Next SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W 096r27379 PCTIU~ ?7 dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 2 hours. The mixture is then exposed to the solid support and mixed for 24 hours. (The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass;
isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkvl chain with terminal carbon activated 0 as free alcohol, bromide etc.)J. The mixture is then diluted with ether, washed with aqueous NaHCO3 (2X), water (2X), and brine (2X) . Next, the compound/support ( 1.0 equivalents) in dimethyl-formamide (.10 Molar) at 25 ~C, is added piperidine ( 1.1 equivalents) . The support is stirred or exposed for 1 hour and is then diluted with ether, and washed with aqueous CuSO4 (2X), water (2X), and brine (2X). The final step affords compound 3040.

SUBSTITUTE SHEET (RULE 26) WO 96127379 ~ 3C~03227 _165-BnO O CO2H BnO O~'o{ Soiid Sui~ort Bn;~XNHFMOC Bn~NH2 BnO BnO

1. DCC, HOBT, Et3N, DMF
2. piperidine, DMF
o BnO~O~o{ Solid Sui~ort ~BnO'~'N
BnO~O~OHBnO O~ ~ ~OBn BnO'~'NHFMOC+H2N~OBn BnO OBn 1. DCC, HOBT, Et3N, DMF
~,~ 2. piperidine, DMF

BnO~O~ Solid Sui-4~ort OBnO- 'N
BnO,~,O~ OH BnO o~rO~--OBn BnO'JyL'NHFMOC N~. OBn BnOBnO~~O~OOBn 3030BnO~L NH2 BnO

1. DCC, HOBT, Et3N, DMF
~ 2. piperidine, DMF

/terate as many steps as needed Scheme 3001 SUBSTITUTE SHEET (RULE 26) CA 022l4789 l997-09-0~
W096~7379 r~ 96/03227 Physical Data for scheme 9.

Phosphoramidate 138 (2 diastereomers): IR, (neat) cm-l:
3089, 2964, 2927, 2856, 2253, 1497, 1455, 1396, 1363, 1253, 1184, 1156, 1094, 1028, 978, 876, 836, 779, 735, lH-NMR (400 MHz, C6D6): ~ 7.34 (m, 5 H, Ph), 7.14 (m, 10 H, Ph), 4.97 (m, 4 H, CH2Ph), 4.78 (m, 2 H, CH2Ph), 4.07-3.24 (m, 13 H, OCH, OCH2, CH2CN), 1.81 (m, 2 H, CH(CH3)2), 1.16 (m, 12 H, CH3CH), 1.03, 1.02 (2 s, 9 H, lBuSi), 0.20, 0.18, 0.16, 0.15, (4 s, 6 H, Me2Si);
HRMS: C43H6307N2PSi, Calc. (M+Cs+): 911.3197; ~ound:
911.3185.

Naphthoylester 135 IR, (neat) cm-l: 3494, 3062, 2919, 1716, 1630, 1600, 1454, 1355, 1284, 1228, 1197, 1091, 779, 736i lH-NMR (250 MHz, CDCI3): ~ 8.58 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7. 89 (m, 2 H, Ar), 7.59 (m, 2 H, Ar), 7.32 (m, 15 H, Ph), 4.95 (m, 3 H), 4.90 (d, ,;r=4 5 Hz, 1 H), 4.69 (m, 3 H), 4.52 (dd, J = 3.9, 12.0 Hz, 1 H), 3.91 (dd J = 2.6, 12.0, 1 H), 3.83 (d, ~J --8.3, 1 H), 3.70 (m, 4 H), 3.96 (m, 1 H), 2.25 (s, 1 H, OH). HRMS: C3gH3807 Calc. (M+Cs+): 751.1672i found:
751.1668.

Dimer 142 IR, (neat) cm-l: 3397, 3030, 2923, 2254, 1718, 1653, 1629, 1497, 1453, 1355, 1284, 1227, 1197, 1094, 1029, 780. lH-NMR (400 MHz, C6D6): ~ 8.82 (s, 1 H, Ar), 8.26 (d, 1 H, Ar), 7.72 (m, 1 H, Ar), 7.61 (m, H, Ar), 7.48 (m, 1 H, Ar), 7.37-6.95 (m, 32 H, Ar, Ph), 4.89-4.18 (m, 21 H, CH2Ph, CH2-Ar, -CH2CH2CN, CHCH2-Ar and CH20H), 3.95-3.45 (m, 13 H, CH- and CH2-sugar), 1.71 (s, 1 H, OH); HRMS: Cl70H7205NP calc. (M+H+):
1198.4718; found: 1198.4715.

SUBSTtTUTE SHEET (RULE 26) CA 022l4789 l997-09-05 W096127379 ~JU~ 3227 Te~ramer 150 IR, (neat) cm-l: 3420, 3064, 2924, 2255, 1721, 1497, 1455, 1357, 1278, 1028, 737. lH-NMR (400 MHz, CDCI3): ~ 8.41 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7.91 (m, 2 H, Ar), 7.55 (m, 2 H, Ar), 7.30 (m, 60 H, Ph), 4.93-4.05 (m, 39 H, CH2Ph, CH2-Ar, CH2CH2CN and CH2OH), 3.88-3.27 (m, 23 H, CH- and CH2-sugar), 2.58 (s, 1 H, OH). HRMS: Cl32Hl40O3lN3P3 Calc. (M+Cs+):
2488.7738; found: 2488.7758.
Tetramer 154 IR, (neat) cm-l: 3376, 2934, 1450, 1244, 1110, 1088. lH-NMR (400 MHz, D20): ~ 8.41 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7.91 (m, 2 H, Ar), 7.55 (m, 2 H, Ar), 4.93-4.05 (m, 4 H, CH2-Ar and CX2OH), 3.88-3.27 (m, 32 H, CH- and CH2-sugar); HRMS: C39H59O3lP3 Calc.
(M+H+): 1117.2331; ~ound: 1117.2350.

SUBSTITUTE S~tEET (RULE 26)

Claims

What is claimed is:

1. An oligomeric carbopeptoid compound comprising carbohydrate amino acid subunits (CA's) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen represented by the following formula:
CA1-(CO-NH)-CA2 wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith and CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.

2. In a process for synthesizing an oligomeric carbopeptoid compound, a coupling step wherein two or more carbohydrate amino acid subunits (CA's) are coupled by means of an amide linkage having a carbonyl carbon and an amido nitrogen for synthesizing said oligomeric carbopeptoid compound, said amide linkage being represented by a formula as follows:
CA1-(CO-NH)-CA2 wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.

3. A library of oligomeric carbopeptoid compounds employable for drug screening, each oligomeric carbopeptoid compound including at least two carbohydrate amino acid subunits (CA's) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen, said amide linkage being represented by the following formula:
CA1-(CO-NH)-CA2 wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.

4. An improved process for synthesizing a library of oligomers, the process employing an elongation step wherein subunits are coupled to one another to produce the oligomers, wherein the improvement comprises:
in said elongation step the oligomer includes at least two carbohydrate amino acid subunits (CA's) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen represented by the following formula:

CA1-(CO-NH)-CA2 wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.

5. A derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons, said anomeric carbon being substituted with a carboxyl radical, each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, blocked amino, differentially protected amino, and hydrogen, with the proviso that at least one radical is a differentially protected amino.

6. A derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons, said anomeric carbon being substituted with a carboxyl radical, each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, blocked amino, unprotected amino, and hydrogen, with the proviso that at least one radical is an unprotected amino and at least one radical is a blocked hydroxyl or amino.

7. An oligomeric carbonucleotoid molecule comprising carbohydrate C-glycoside subunits (CG's) coupled to one another via a phosphodiester linkage represented by the following structure:

CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;

CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.

8. In a process for synthesizing an oligomeric carbonucleotoid molecule, a coupling step wherein two or more carbohydrate C-glycoside subunits (CG's) are coupled by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:
CG1-C1'-(O-PO(OH)-O)-CG2 wherein.
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.

9. A library of oligomeric carbonucleotoid molecules employable for drug screening, each oligomeric carbonucleotoid molecule including at least two carbohydrate C-glycoside subunits (CG's) coupled to one another by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:

CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.

10. An improved process for synthesizing a library of oligomers, the process employing an elongation step wherein subunits are coupled to one another to produce the oligomers, wherein the improvement comprises:
in said elongation step the oligomer is a carbonucleotoid including at least two carbohydrate C-glycoside subunits (CG's) are coupled by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:
CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.

11. A derived carbohydrate C-glycoside having an anomeric carbon and non-anomeric carbons, said anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to an activated phosphite, each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, differentially protected hydroxyl, and hydrogen, with the proviso that at least one radical is a differentially protected hydroxyl.

12. A derived carbohydrate C-glycoside having an anomeric carbon and non-anomeric carbons, said anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to an activated phosphite, each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, unprotected hydroxyl, and hydrogen, with the proviso that at least one radical is an unprotected hydroxyl and at least one radical is a blocked hydroxyl.