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Université René Descartes – Paris 5 UFR Biomédicale des Saints-Pères Ecole Doctorale du Médicament. Strategic investigations for the design of a library of liposidomycins analogs, natural antibiotics dedicated to the MraY translocase. Maryon GINISTY Direction : Pr. Yves Le Merrer - PowerPoint PPT PresentationTRANSCRIPT
Université René Descartes – Paris 5 UFR Biomédicale des Saints-PèresUFR Biomédicale des Saints-Pères
Ecole Doctorale du MédicamentEcole Doctorale du Médicament
Strategic investigations for the Strategic investigations for the design of a library of design of a library of
liposidomycins analogs, natural liposidomycins analogs, natural antibiotics dedicated to the MraY antibiotics dedicated to the MraY
translocasetranslocase
Maryon GINISTY
Direction : Pr. Yves Le Merrer
Laboratoire de Chimie et Biochimie Pharmacologiques et ToxicologiquesDirection : Dr. Daniel Mansuy - UMR 8601 – CNRS45, rue des Saints-Pères - 75270 Paris Cedex 06- France
2
ANTIBACTERIAL RESISTANCE : ANTIBACTERIAL RESISTANCE : A MAJOR OBSTACLE FOR A MAJOR OBSTACLE FOR
ANTIBIOTHERAPYANTIBIOTHERAPY 1940’s : development of penicillin and appearance of the concept of antibiotics
« Agents with specific antibacterial action and with toxicity selectively directed against bacteria in low concentrations »
► Bacteriostatic effect (decrease or stop of bacterial growth)
► Bactericid effect (destruction of bacteria)
● Complexity and adaptability of bacterial world
► Therapeutic failure► Development of a large number of antibiotics classified according to various
criteria : site of action, origin, administation route, structure
⇒ Eight major families : -lactams, aminosides, macrolides, sulfamides, poly- et glyco-peptides, cyclins, (fluoro)quinolons…
3
● Two types of resistance :
► Natural resistance (intrinsic property related to the bacterial genetic program)
► Acquired resistance (property resulting from genetic modifications of the bacterial cells)
● Five major mechanisms of resistance :
- Overproduction of antibiotic target- Metabolic bypass of inhibited reaction - Inactivation of antibiotic by enzymatic modification - Modification of target eliminating or reducing antibiotic
binding to target
RESISTANT STRAINS AND MECHANISMSRESISTANT STRAINS AND MECHANISMS
⇒ Resistant strain : strain able to develop in the presence of an antibiotic concentration notably higher than that which inhibits development of other strains of same species
antibiotic « modifying »enzyme
modified antibiotic
antibiotic
X
modifiedreceptor
antibiotic
receptor resistancegene
pump
- Decrease of cellular permeability to antibiotic
4
⇒ Four sites of action specific to procaryote bacterial cells
- ribosomes responsible for protein synthesis
- metabolism of nucleic acids⇒ inhibition of DNA synthesis⇒ inhibition of DNA transcription into messenger RNA
- oxydoreduction (5-nitro-imidazoles) via formation of superoxides and nitro radicals responsible of irreversible damage on bacterial DNA- cell wall biosynthesis
SITES OF ACTION OF ANTIBIOTICS AND SITES OF ACTION OF ANTIBIOTICS AND POTENTIAL TARGETSPOTENTIAL TARGETS
Gram (-) cell
Gram (+) cell lipopolysaccharide
periplasm
cytoplasmic membranecytoplasmic membrane
external membrane
peptidoglycancytoplasmcytoplasm
mRNA
ribosomeDNA
DNA-gyrase
RNA-polymerasemRNA
Bacterial wall
N
N
R
CH3O2N
5-nitro-imidazoles
BACTERIA
aminoacid
5
OHN
O
NH2
D-Cycloserin
PERIPLASM
ANTIBIOTICS AND BACTERIAL
PEPTIDOGLYCAN BIOSYNTHESIS
UDP-GlcNAc
UDP-GlcNAc-enolpyruvate
UDP-MurNAc
UDP-MurNAc-L-Ala
UDP-MurNAc-dipeptide
UDP-MurNAc-tripeptide
PEP
NADPH
L-Ala D-Ala
D-Glu L-Glu
meso-A2pm
D-Ala-D-Ala D-Ala
MurA
MurB
MurC
MurD
MurE
MurF
Alr
MurI
Ddl
-O2C OPO32-
PEP
PO32-
O
Fosfomycin
UDP-MurNAc-pentapeptide
bacitracin
vancomycinmoenomycinpenicillin
cephalosporin
D-cycloserin
D-cycloserin
fosfomycin
tunicamycinmuraymycinmureidomycinliposidomycin
MraY
O
O-
NH3+
D-Alanine
CYTOPLASM
UMP
PiUDP-GlcNAc
UDP
BacA MurG
PBPs
PBPs
Lipid I
Lipid II
AcceptorPolymer
Peptidoglycan
Undecaprenyl-PP
Undecaprenyl-P
MEMBRANE
N-acetylmuramic acid
N-acetylglucosamine
tétrapeptide
pentapeptide
O
O
NHCOCH3
OO
O
NHCOCH3
CHH3C
C
NH
O
tetrapeptide
N-acetylmuramic acid N-acetylglucosamine
6
INHIBITORS AND NATURAL SUBSTRATE INHIBITORS AND NATURAL SUBSTRATE OF MraY TRANSLOCASEOF MraY TRANSLOCASE
OO
OH
HO
NH
CH2
OOH
NHN
O
OO
OHOHO
HOAcHN
HO OH
Tunicamycins
n
A : n=9
B : n=10
C : n=8
D : n=11
OOH2N
HO OCH3
O
HO OH
NH
N
O
O
CO2HHN
HN
NH
HNHO2C
O
OOR
HN
NH
HN
O
Muraymycins
A1: R=COC11H22N(OH)C(NH2)=NH
A3: R=COC11H22NHC(NH2)=NH
C1: R=H
NHN
O
O
O
OH
O
HN
CH3
N
NH
R1
O
HO
HN
ONH
R2
HN
O
R3
OH
O
H3C
R1=H, glycinylR2= CH3S(CH2)2-R3= m-hydroxyphenyl
Mureidomycins
O
NHN
O
ON
N
CH3
HO2C
O
O
CH3R
O
O
HO2C
O
O
O
NH2
A: R=
B: R=
C: R=
Liposidomycins
OH OH
OHHO3SOO
OH
HOO
AcHN
O
pentapeptide-HNO
PO
PO
-O O
O
HO OH
N
NH
O
O
O O-
Me
UDP-Mur-NAc-pentapeptide
7
N
N
CH3
HO2C O
CH3
12
345
67
N
N
CH3
HO2C O
CH3
12
345
67
O O
OHH O3S O
NH2
O O
OHH O3S O
NH2
NHN
O
O
O
HO OH
H1'
2'3'4'
STRUCTURE OF LIPOSIDOMYCINSSTRUCTURE OF LIPOSIDOMYCINS
A: R=
B: R=
C: R=
O
R
O
O
HO2C
O
5'NHN
O
O
O
HO OH
H1'
2'3'4'
S
S
S S
8
N
NCH3
CH3O
PhCOO
O
EtON Boc
CH3
PhCOO
O
EtON H
CH3
PhCOO
O
EtON
N CH3
H3C O
PhCOO
O
EtO
Z
N
N CH3
H3C O
PhCOO
O
EtO
O
Z
SYNTHETIC APPROACHES DESCRIBED IN SYNTHETIC APPROACHES DESCRIBED IN LITTERATURELITTERATURE
OHCacrolein
N
HO
O
CH3Z
N-Z-sarcosine
EtO2C NBoc
CH3
N-Boc-sarcosine ethyl ester
⇖
⇖⇖
1,4-diazepan-2-one moiety
N
NCH3
CH3O
PhCOO
O
EtO 1 23
456
7
Knapp et coll. Tetrahedron Lett. 1992, 33, 5485.Knapp et coll. J. Org. Chem. 2001, 66, 5822.
9
BnO
NH
OMPM
OTBDMSO
OBn
R
N3BnO
NH
OH
OTBDMSO
OBn
R
N3BnO
NH
O
OTBDMSO
OBn
R
N3
Ribosyl-diazepanone Isono et coll. Heterocycles 1992, 34, 1147.
OMPM
OTBDMSN3
BnO
(S1)
HO2C
N3 OBn
(S2)
O OCH3
O O
OH
OH
OMPM
OH
RHO
OtBuOOH, Ti(OiPr)4
L-DET
78%
OMPM
OH
OD-DET
tBuOOH, Ti(OiPr)4
88%
OMPM
OHN3
HO
50%
NaN3
RHO
N3 OH
RHO
NaN3
O OCH3
O O
R :
NH
N
O
BnO OBn
TBDMSO O OCH3
O O
O OCH3
O O
OHC
DCC / HOBt
NH
N
O
BnO OBn
TBDMSO O OCH3
O O
Pd/C, H2
36%
SYNTHETIC APPROACHES DESCRIBED SYNTHETIC APPROACHES DESCRIBED IN LITTERATUREIN LITTERATURE
10
O O
OTBDMSO
HO2C
OH
N3
H O O
OTBDMSO
HO2C
OH
N3
H
NHCH3
PhCOO
CO2Et
O O
OTBDMSO
OH
NH2
O
O NCH3PhCOO
CO2EtEEDQH
NHCH3
PhCOO
CO2Et
O O
OTBDMSO
OH
N3
O
NCH3PhCOO
CO2EtEEDQH
NHCH3
PhCOO
CO2Et
O O
OTBDMSO
OH
N3
O
O NCH3PhCOO
CO2EtEEDQH
Nucleosidyl-diazepanone Knapp et coll. Org. Lett. 2002, 4, 603.
O O
OHO
O H
OH
O O
OTBDMSO
HO
O
O
OTBDMSO
N
N
OH3C
CH3
EtO2C
OHH
O
O
OHHO
NH
N
O
ON
N
OH3C
CH3
EtO2C
OHdeprotection, acylation
thenglycosylation
1/ Oxidation
2/ NaN3
1/ Ozonolysis
2/ Azidereduction
3/ Reductive amination
SYNTHETIC APPROACHES DESCRIBED SYNTHETIC APPROACHES DESCRIBED IN LITTERATUREIN LITTERATURE
11
O
NH
N
O
O
OO
O O
NH
N
O
O
OO
MeO2C
O
BocHN
O
O
O
NH
N
O
O
OO
CbzHN
H
O
TBSO
TBDPSO
O
O
HN
O
BocHN
O
O
O
NH
N
O
O
OO
CbzHN
H
O
TBSO
TBDPSO
H2C
O
HN
O
BocHN
O
O
O
NH
N
O
O
OO
CbzHN
H
O
HO2C
O
NH
N
O
O
OO
CbzHN
H
OH
MeO2C
N
NHO
HO2C
Me
Me
O
O
NH
N
O
O
OHHO
H
O
O
H2NHO
HO
1 Wittig
Nucleosidyl-ribosyl-diazepanone Angew. Chem. Int. Ed. 2005, 44, 1854.
N
NHO
HO2C
Me
Me
O
O
NH
N
O
O
OHHO
H
O
O
H2NHO
HO
N
NHO
HO2C
Me
Me
O
O
NH
N
O
O
OHHO
H
O
O
H2NHO
HO
2 glycosylation
O
N3
O O
F
N
NHO
HO2C
Me
Me
O
O
NH
N
O
O
OHHO
H
O
O
H2NHO
HO
3 peptide couplingTBSO
NHMe
OTBDPS
H2C
N
NHO
HO2C
Me
Me
O
O
NH
N
O
O
OHHO
H
O
O
H2NHO
HO
reductiveamination
4
SYNTHETIC APPROACHES DESCRIBED SYNTHETIC APPROACHES DESCRIBED IN LITTERATUREIN LITTERATURE
12O
HO
OH
AcHNO
pentapeptide-HN
OO-O
PO O
O-O
PO
Lipid I
8
N
N
OHO2C
O
O
R
O
HO2C
O
O
HO OH
NH
N
O
O
OO(SO3)H
OH
H2NLiposidomycins
OHO
OH
AcHNO
pentapeptide-HN
OO-O
PO O
O-O
PO
O
HO OH
NH
N
O
O
UDP-N-acetyl-muramoylpentapeptide
STRUCTURE-ACTIVITY RELATIONSHIP : STRUCTURE-ACTIVITY RELATIONSHIP : DEVELOPMENT OF A NEW DEVELOPMENT OF A NEW
PHARMACOPHOREPHARMACOPHORE
O
OP
O-O- 8
Undecaprenylphosphate
OHO
OH
AcHNHO
O OHN
R
O
HOOH OH
O
HO OH
NH
N
O
O
Tunicamycins
MraY
13
N
N
O
CH3
HO2C
OO
OHHO
O
NH2
H3C
O
HO OH
NHN
O
ON
N
O
CH3
HO2C
OO
OHHO
O
NH2
H3C
O
HO OH
NHN
O
ON
N
O
CH3
HO2C
OO
OHHO
O
NH2
H3C
O
HO OH
NHN
O
ON
N
O
CH3
HO2C
OO
OHHO
O
NH2
H3C
O
HO OH
NHN
O
O
O
O
HO
HO
NH2
O
HO OH
NHN
O
O
O
O
HO
HO
NH2
O
HO OH
NHN
O
O
O
OHHO
O
NH2
**
NH
N
O
R2
R3O
R4O**
NH
*
N
R1
O
R2
R3O
R4O
O
OHHO
O
NH2
**
NH
N
O
R2
R3O
R4O
O
OHHO
O
NH2
NH
N
O
R2
O
O
O
O
HO
HO
NH2
O
HO OH
NHN
O
O
STRUCTURE-ACTIVITY RELATIONSHIP: STRUCTURE-ACTIVITY RELATIONSHIP: DEVELOPMENT OF A NEW DEVELOPMENT OF A NEW
PHARMACOPHOREPHARMACOPHORE
Target scaffold
R2= NHN
O
O
Pharmacophore structure
Structure of natural molecules
14
SCAFFOLD RETROSYNTHESISSCAFFOLD RETROSYNTHESIS
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'2'3'
4'5'
1''
H2N OH
PO
amino-dihydroxy-butane
Y
NH2HO2C
OHL-Serine
O
PO OP
H2N1
23
45
X
5-amino-ribose
O
HO OH
OHO
HO
L-ascorbic acid
O
HO OH
HO1
23
45
OH
D-ribose
P: protecting groupsX: halogen
N-ALKYLATION
O-GLYCOSYLATION
PEPTIDECOUPLING
⇒
15
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
NH2
PO
H2N
O OH
HO
L-serine
HO
amino-dihydroxy-
butane
Y
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
AO
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
STRATEGIES TOWARDS SCAFFOLD STRATEGIES TOWARDS SCAFFOLD SYNTHESISSYNTHESIS
O
PO OP
H2N
O O
H2N 1'
2'3'
4'5' A
HOB
HN
NH
O
PO
OH
PO
1
2
3
4
5
67
C B
NH2
POamino-dihydroxy-butane
HO Y
O
PO OP
H2N1
23
45
X
5-amino-ribose
A
O
PO OP
H2N
1
23
45
X
5-amino-ribose
H2N
O OH
HO L-serine
GLYCOSYLATION
N-ALKYLATION
PEPTIDE COUPLING
N-ALKYLATION
PEPTIDE COUPLING
GLYCOSYLATION
16
O O
P2O OP2
R1HN
NHR3
R2O2C
O O
P2O OP2
R1HN
NHR3
R2O2Caminedeprotection
FunctionalizedL-serinyl-O-ribofuranoside
O O
P2O OP2
R1HN
NHR3
R2O2Caminedeprotection
FunctionalizedL-serinyl-O-ribofuranoside
O O
P1O OP1
P1O
NHR3
R2O2C
aminedeprotection
O O
P1O OP1
P1O
NHR3
R2O2C
O
P1O OP1
R1HN
O
NHN3
HO2C
HO
TBDPSO
O
P1O OP1
R1HN
O
NHN3
HO2C
HO
TBDPSO
ACCESS TO THE SCAFFOLD BY « CHAIN ACCESS TO THE SCAFFOLD BY « CHAIN EXTENSION »EXTENSION »
STEPS AND PRECURSORS
O
P2O OP2
R1HN
O
NH
HN
HO
O
TBDPSO
O
HO OH
H2N
O
NR2
HN
R1O
O
HO
scaffold
O O
P2O OP2
R1HN
NH2
R2O2C
N3
PO
Oazido-epoxideO O
P1O OP1
P1O
NHR3
R2O2C
functionalizationof ribosyl moiety
N-alkylation
OO
P1O OP1
P1O
functionalization of ribosyl moiety
NHN3
R2O2C
POPO
O O
P1O OP1
P1O
NH2
R2O2C
N3
PO Oazido-époxyde
N-alkylation
⇗
⇘
O X
P2O OP2
R1HN
NHR3
R2O2Cglycosylation
OH
prefunctionalizedribofuranose
O X
P1O OP1
P1O
NHR3
R2O2CglycosylationOH
O X
P1O OP1
P1O
NHR3
R2O2CglycosylationOH
STRATEGY 1
STRATEGY 1
STRATEGY 2
17
OOH
"donor" sugarO
OR
OX
H+
R-OH
acceptor
X= Br, Cl, F, SR
= OCOR, O3SR, OP(OR)2, OPO2-OR'.
R-OH (acceptor)
Substitution via activation of anomeric position
Direct acid-catalyzed substitution
GLYCOSYLATION STEPX= Br, Cl, F
ACCESS TO THE SCAFFOLD BY « CHAIN ACCESS TO THE SCAFFOLD BY « CHAIN EXTENSION »EXTENSION »
18
⇒ Tricky step :
- for the formation of O-glycosidic derivatives, less known than that of N-glycosidic analogs
- in the particular case of threonyl and serinyl acceptors
"H"
O O
PO OP
NHR2
YOR1
O
H
HO
NHR2
OR1
O
O
PO OP
Y
"H"
O O
PO OP
NHR2
YOR1
O
H
HO
NHR2
OR1
O
O
PO OP
Y
ELABORATION OF ELABORATION OF OO-GLYCOSYLATION STEP (1) -GLYCOSYLATION STEP (1)
O X
P1O OP1
Y
O O
P1O OP1
Y
CO2P2
NHP3HOCO2P2
NHP3+
R
R = H : L-serineR = CH3 : L-threonine
R
O O
PO OP
R2HN H
YOR1
O
B
O O
PO OP
NHR2
YOR1
OO O
PO OP
Y
NHR2
OR1
O
H2OO OH
PO OP
Y
Basic lability
Acid lability
O X
P1O OP1
Y
O OR
P1O OP1
Y+ R-OH
19
O
OPO
R
O
PO
H
O X
P1O OP1
Y
O O
P1O OP1
Y
CO2tBu
NHP3HOCO2tBu
NHP3+
P3 = Boc, Cbz, Fmoc
O X
P1O OP1
Y
O O
P1O OP1
Y
CO2tBu
NHP3HOCO2tBu
NHP3+
O X
P1O OP1
Y
O O
P1O OP1
Y
CO2tBu
NHP3HOCO2tBu
NHP3+
⇒ Success of the reaction and control of stereochemistry depending on three principal factors :
- nature of glycosylation activator
- nature of activation in anomeric position
-nature of the C-2 substituent of the ribose controling - or -selective introduction of serine (anchimeric assistance)
activator
O
OPO
R
O
PO
X O
O
O
H
PO
PO
RX
R'-OH
H
O
ROCO
OR'
PO
PO
anomer
ELABORATION OF ELABORATION OF OO-GLYCOSYLATION STEP (2) -GLYCOSYLATION STEP (2)
20
O OP1
P2O OP2
Y
O
P2O OP2
Y
Xglycosylation
reagent
HO CO2tBu
NHP3
OO
P2O OP2
Y
CO2tBu
NHP3
R1= HR2 = Bn
Koenigs-Knorr method
R1= AcR2 = Ac, Bz, Bn
O R
O O
Y
R = OH, FY = N3, PhtN, NHZ
R1, R2 = H
O X
RO OR
RO
R = Ac X = Br, ClR = Bz X = BrR = Bn X = Br
ACTIVATION IN ANOMERIC POSITIONACTIVATION IN ANOMERIC POSITION
O OR1
R2O OR2
R2O
X = Cl
SOCl2, DCM,
0°C to RT
X = Br
TMS-Br, DCM, -40°C to RT.
STRATEGY 1 O F
BnO OBn
BnO
DAST, THF, -30°C to RT, 1h.95%
(= 99/1)
STRATEGY 2
O OAc
AcO OAc
N3
21
O OMe
O O
N3
HN3* (2,8M), PPh3,DIAD, THF, 0°C.
98%
O OMe
O O
HO
Me2CO, MeOH, HCl(g),RT.
75%
O OMe
O O
PhtN
PhtNK, HMPT,120°C, 24h.
74%
O OMe
O O
TsO
TsCl, Et3N,DMAP, CH2Cl2,
0°C to RT.
84%
O OMe
O O
HO
1/ DOWEX 50W-H+, MeOH, 65°C, 17h.2/ Ac2O, pyridine, RT, 2h.
O OMe
AcO OAc
N3
O OH
O O
ZHN
a 10%
O OMe
O O
ZHN
ZCl, DIEA,CH2Cl2, RT,
2h.100 %
Pd/C, H2, EtOH, CHCl3, TA, 1h30.
98%
O OMe
O O
ClH3N
Pd/C, H2, EtOH, CHCl3, RT, 1h30.
98%
FORMATION OF PREFUNCTIONALIZED FORMATION OF PREFUNCTIONALIZED RIBOFURANOSIDESRIBOFURANOSIDESO OH
HO OH
HO D-Ribose
* 2 NaN3 + H2SO4 2 HN3 + Na2SO4H2O
0°C
50%(= 3,4)
O OAc
AcO OAc
N3
AcOH, Ac2O,H2SO4,RT, 2h.
35%
O OH
O O
N3
40%a
O OH
O O
PhtN
a 25%
a : 1/ H2SO4 (0,1N), 65°C, 4h; 2/ Me2C(OMe)2, CSA, Me2CO, 50°C, 30 min.
22
R= Boc, Cbz, FmocR= BocR= CbzR= Fmoc, Cbz
SYNTHESIS OF SYNTHESIS OF LL-SERINYL ACCEPTORS-SERINYL ACCEPTORS
BnOCO2H
NHBoc
BnOCO2tBu
NHBoc
Cl3C OtBu
NH
BF3.OEt2,cyclohexane, CH2Cl2,
TA, 17 h.
96%
HOCO2tBu
NHBoc
H2, Pd(OH)2/ C,
EtOH abs., AcOH,TA, 72 h.
99%
HOCO2H
NH2
L-serine
HOCO2tBu
NHR
N-carbamoyl-L-serine tert-butyl ester
HOCO2H
NHR
tBu-Br, BnEt3NCl,
K2CO3, CH3CN,
50°C, 24h.
88%
HOCO2tBu
NHZR= Cbz
Cl3C OtBu
NH
AcOEt,cyclohexane,
20°C, 24h.
84%
HOCO2tBu
NHFmoc
R= Fmoc
23
O OP2
OP1P1O
Y
O X
OP1P1O
Y
O O
OP1P1O
Y
CO2tBu
NHP3
HOCO2tBu
NHP3
activation glycosylation STRATEGY 1 (riboses not functionalized )
STRATEGIE 2 (prefunctionalized riboses)
PP11 PP22 YY ActivationActivation XXX = Br, Cl, FX = Br, Cl, F
ZZ
AgOTf, DCM, -15°C, 15 h.AgOTf, DCM, -15°C, 15 h.
: 100%: 100% 3232
ZZ : 100%: 100% 9292
BocBoc 0,30,3 6969BocBoc 0,30,3 6969
AcAc AcAc OAcOAc
TMSBr, DCM, -40°C à TATMSBr, DCM, -40°C à TA
BrBr
BzBz AcAc OBzOBz BrBr
BnBn AcAc OBnOBn BrBr
CMeCMe22 HH NN33
DAST, THF, -30°C à TA, DAST, THF, -30°C à TA,
2h.2h.
FF
CMeCMe22 HH NN33 FF
BnBn HH OBnOBn FF
O OH
OO
Y
O F
OO
Y
O O
OO
Y
CO2tBu
NHP
DAST, THF, -30°C à TA, 1h.
HO CO2tBu
NHP
activateur
SnCl2/ AgClO4
BocBoc
SnClSnCl22, AgClO, AgClO44, -15°C à TA, , -15°C à TA,
48 à 72 h.48 à 72 h.
2,152,15 6464
FmocFmoc 1,71,7 100100
BocBoc 1,21,2 4444
HOCO2tBu
NHP
O OAc
RO OR
RO
O
RO OR
ROX O O
RO OR
RO
CO2tBu
NHP
activator
SELECTION OF ACTIVATORS AND OPTIMIZATION SELECTION OF ACTIVATORS AND OPTIMIZATION OF GLYCOSYLATION CONDITIONSOF GLYCOSYLATION CONDITIONS
Ginisty M., Gravier-Pelletier C., Le Merrer Y., Tetrahedron: Asymmetry 2004, 15, 189-193.Ginisty M., Gravier-Pelletier C., Le Merrer Y., Tetrahedron: Asymmetry 2006, 17, 142-150 .
Hg(CN)2
AgClO4
TMS-OTf BF3.OEt2
AgOTf SnCl2/ AgClO4
PP33 GlycosylationGlycosylationRatio Ratio
))Yield Yield (%)(%)
24
O O
P1O OP1
P1O
NHR3
R2O2C
O O
P2O OP2
R1HN
NH2
R2O2C
N3
PO O
azido-epoxide
O X
P2O OP2
R1HN
NHR3
R2O2C
glycosylation OH
prefunctionalizedribofuranose
O X
P1O OP1
P1O
NHR3
R2O2C
glycosylationOH
O O
P2O OP2
R1HN
NHR3
R2O2C
functionalizedL-serinyl-O-ribofuranoside
O O
P1O OP1
P1O
NHR3
R2O2C
O
P1O OP1
R1HN
O
NHN3
HO2C
HO
TBDPSO
O O
P1O OP1
P1O
NH2
R2O2C
N3
PO Oazido-epoxide
⇗
⇘
O X
P1O OP1
P1O
NHR3
R2O2Cglycosylation OH
STRATEGY 1
STRATEGY 1
STRATEGY 2
aminedeprotection
aminedeprotection
functionalization of ribosyl moiety
ACCESS TO THE SCAFFOLD BY « CHAIN ACCESS TO THE SCAFFOLD BY « CHAIN EXTENSION »EXTENSION »
25
PhtNK, DMF,
160°C, 12h.O O
O O
PhtN
tBuO2C
NHZ
1'
2'3'
4'5'
1 2 3
O OHO
HO OH
1'
2'3'
4'5'
12
3tBuO2C
NHR
CH3C(OMe)2, MeOH, APTS, (CH3)2CO. O O
HO
O O
1'
2'3'
4'5'
12
3tBuO2C
NHR
O O
AcO OAc
AcO
tBuO2C
NHZ
O O
HO OH
HO
tBuO2C
NHZK2CO3, MeOH/ H2O,
TA, 1h.
70%
1'
2'3'
4'5'
12 3 1
2 3
1'
2'3'
4'5'
O O
O O
HO
tBuO2C
NHZ
1'
2'3'
4'5'
1 2 3
O O
BzO OBz
BzO
tBuO2C
NHZ
O O
HO OH
HO
tBuO2C
NHZK2CO3, MeOH/ H2O,
TA, 1h.
1'
2'3'
4'5'
12 3 1
2 3
1'
2'3'
4'5'
O O
BzO OBz
ZHN H
BzOOtBu
O
O O
BzO OBz
BzO
NHZ
OtBu
O
+O OBnO
BnO OBn
1'
2'3'
4'5'
12
3tBuO2C
NHBoc
O OHO
HO OH
1'
2'3'
4'5'
12
3tBuO2C
BocHN
Pd(OH)2/C, H2, EtOH abs., CH3CO2H,
TA, 24h.
100%
TsCl, DMAP, Et3N, CH2Cl2,
0°C to TA, 6h.
81%
O O
O O
TsO
tBuO2C
NHZ
1'
2'3'
4'5'
1 2 3
O O
HO OH
HO
NHR3
R2O2C
1'
2'3'
4'
5'
12 3
FUNCTIONALIZATION OF RIBOSYL MOIETYFUNCTIONALIZATION OF RIBOSYL MOIETY
O O
P2O OP2
R1HN
NHR3
R2O2C
1'
2'3'
4'5'
1 32
O O
P2O OP2
HO
NHR3
R2O2C
1'
2'3'
4'
5'
12 3
O O
P1O OP1
P1O
NHR3
R2O2C
1'
2'3'
4'5'
12 3
functionalizationat C-2’ and C-3’
positions
substitutionof the
5’-OH function deprotection
C-2’ and C-3’protection
substitutionof the5’-OH
function
P1 = Ac, Bz, BnP2 = C(CH3)2
OMe
X
MeO
MeOH
O OH
BzO OBz
BzO
MeOCO2tBu
NHZ
major product
+R = Z 82%
Boc -
26
O O
P2O OP2
R1HN
NH2
R2O2C
O
P1O OP1
R1HN
O
NHN3
HO2C
HO
TBDPSO
O O
P1O OP1
P1O
NHR3
R2O2C
O X
P2O OP2
R1HN
NHR3
R2O2C
glycosylation OH
prefunctionalizedribofuranose
O X
P1O OP1
P1O
NHR3
R2O2C
glycosylationOH
O O
P2O OP2
R1HN
NHR3
R2O2C
functionalizedL-serinyl-O-ribofuranoside
O O
P1O OP1
P1O
NHR3
R2O2C
O O
P1O OP1
P1O
NH2
R2O2C
⇗
⇘
O X
P1O OP1
P1O
NHR3
R2O2Cglycosylation OH
STRATEGY 1
STRATEGY 1
STRATEGY 2
aminedeprotection
aminedeprotection
functionalizationof the ribosyl moiety
N3
PO O
azido-epoxide
N3
PO Oazido-epoxide
ACCESS TO THE SCAFFOLD BY « CHAIN ACCESS TO THE SCAFFOLD BY « CHAIN EXTENSION »EXTENSION »
27
O O
O O
H2N
tBuO2C
NHFmoc
1'2'3'
4'5'
21
3
O O
O O
H2N
tBuO2C
NH2
1'2'3'
4'5'
21
3
HCO2NH4
Pd/C 10%
MeOH, TA.
O O
O O
N3
NH2
tBuO2C
N3
PO
O
O O
O O
N3
N3
HO
PO
tBuO2C
NH
azidoreduction
O O
O O
H2N
tBuO2C
NHFmoc
1'2'3'
4'5'
21
3
O O
O O
Y
tBuO2C
NHFmocprotection
of C5'-amino group
1'2'3'
4'5'
21
3
O O
RO OR
RO
tBuO2C
NHZ
O O
RO OR
RO
tBuO2C
NH2
1'2'3'
4'5'
21
3
1'2'3'
4'5'
21
3
AMINE DEPROTECTIONAMINE DEPROTECTION
STRATEGY 1
H2, Pd(OH)2/ C, CH3CO2H, EtOH abs., RT, 24h.
H2, Pd(OH)2/ C, CH3CO2H, EtOH abs., RT, 48h.
H2, Pd black, CH3CO2H, RT, 48h.
R = Ac, Bz
X
STRATEGY 2
O O
O O
N3
tBuO2C
NHFmoc
1'2'3'
4'5'
21
3
Y= PhtN-, ZHN-
O O
O O
Y
tBuO2C
H2N
1'2'3'
4'5'
21
3deprotection
of C2-amino group
X
28
O OY
O O
tBuO2C
NH2
1'2'
4'5'
1 2 3
A
B
3'
Powerful glycosylation conditions for the diastereoselective formation of Powerful glycosylation conditions for the diastereoselective formation of serinyl-5’-amino-serinyl-5’-amino--D-ribofuranoside derivatives-D-ribofuranoside derivatives
⇒ ⇒ unfinished strategy because of difficult functionalization of the unfinished strategy because of difficult functionalization of the ribosyl moiety and amine deprotection. ribosyl moiety and amine deprotection.
PerspectivePerspective : : ⇒ ⇒ strategy 2 : glycosylation of 2,3-strategy 2 : glycosylation of 2,3-OO-isopropyliden--isopropyliden-DD-ribofuranoside -ribofuranoside derivatives differently derivatives differently NN-protected, -protected, whose synthesis was already carried out. whose synthesis was already carried out.
. .
O OMeN3
O O
123
45O OH
HO
HO OH
123
45
functionalizationof the
ribosyl moiety O XY
O O
123
45
ACCES TO THE SCAFFOLD BY « CHAIN ACCES TO THE SCAFFOLD BY « CHAIN EXTENSION » : CONCLUSION AND EXTENSION » : CONCLUSION AND
PERSPECTIVESPERSPECTIVES
1) glycosylation
2) aminedeprotection
HOCO2tBu
NHFmoc
123
Y = PhtN-, ZHN-X = activated group
29
ACCESS TO THE SCAFFOLD BY DIRECT COUPLINGACCESS TO THE SCAFFOLD BY DIRECT COUPLING
O
PO OP
HN
NH
O
PO
O
PO
H2N
1
2
3
45
67
1'
2'3'
4'5'
1''
C B
A
O
PO OP
H2N
O O
H2N 1'
2'3'
4'5' A
HO
HN
NH
O
PO
OH
PO
1
2
3
4
5
67
C B
O
PO OP
H2N1'
2'3'
4'5'
X
5'-amino-ribose
NH2
PO
H2N
O OH
HO
L-sérine
HO
amino-dihydroxy-
butane
Y
O
PO OP
H2N
1'
2'3'
4'5'
X
5'-amino-ribose
NH2
O OH
HO L-serine
GLYCOSYLATION
N-ALKYLATION
PEPTIDECOUPLING
GLYCOSYLATION
NH2
POamino-dihydroxy-butane
HO Y
30
1,4-diazépan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
1,4-diazépan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
1,4-diazepan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
NH2
PO
amino-butanol
PO
Y
O OHO
HO
HO OHL-ascorbic acid
NH
H2N
O
PO
OH
1
2
34
5
6 7
Y
PO
N-alkylation
peptidecoupling
N-alkylationNH2
CO2H
NHPO
PO
OH
12
34
5
6
7
peptidecoupling
H2N
O
OH
HO
L-serine
CAG STRATEGY
ACGSTRATEGY
ACCESS TO THE SCAFFOLD BY DIRECT COUPLINGACCESS TO THE SCAFFOLD BY DIRECT COUPLING
31
CO2Et
OHO
O OTBDPS
N3O
O
azido-acetonide
FORMATION OF NFORMATION OF N11-C-C22 LINKAGE BY PEPTIDE LINKAGE BY PEPTIDE COUPLINGCOUPLING
- FIRST STEP OF THE CAG STRATEGY -- FIRST STEP OF THE CAG STRATEGY -
SYNTHESIS OF AMINO-BUTANOL PRECURSORS
O
OTBDPS
NH2
OTBDPS
NH2O
O
NH2TBDPSO
EtO2C
amino-epoxide
amino-acetonide
amino-ester
OTBDPS CO2Et
OTBDPSN3
TBDPSO
azido-ester
O
OTBDPS
N3
azido-epoxide
CO2Et
OTBDPSO
O
O
OHHO
OHO
HO
L-ascorbic acid3,4-O-methylethyliden-L-threonine ethyl ester
32
N3 OP
YPO 123
4
+ -1) H2O2, H2O, K2CO3,
0°C, 2h.2) EtI, CH3CN,
85°C, 12h.
78% O OH
CO2EtO
O OH
CO2EtO
23
14O
HO OH
OHO
HO
O
HO OH
OO
O
Me2C(OMe)2, Me2CO, HCl(g), RT, 12h.
94%
L-ascorbic acid
CO2Et
OTBDPSN3
TBDPSO
OTBDPS
N3O
O
PO OP
OPPO 123
4
PO X
OPPO 1234
SN en C2
X OP
OPPO 1234
SN en C3
180° rotation
introduction in C3 position
of the azido group
introduction of the azido
group in C2 position
O N3
OP123
4
N3 OP
OPO
R
1
23
4
P : protecting groupR = OEt, H
introduction of an electrophilic group
in C4 position
Introduction of an electrophilic group
in C1 position
180° rotation
FORMATION OF NFORMATION OF N11-C-C22 LINKAGE BY PEPTIDE LINKAGE BY PEPTIDE COUPLINGCOUPLING
- FIRST STEP OF THE CAG STRATEGY -- FIRST STEP OF THE CAG STRATEGY - SYNTHESIS OF AMINO-BUTANOL PRECURSORS
33NH2O
O OTBDPSPPh3, H2O, THF,
RT, 72h.
78%
O N3
O OTBDPS
TBDPSCl, ImH,DMF,
0°C to RT, 12h.
100 %
O OTBDPS
CO2EtO
O OH
CO2EtO CO2Et
OTBDPSN3
TBDPSO1
234
TFA, H2O, THF, 0°C, 3h.
74%
HO OTBDPS
CO2EtHO TBDPSCl, ImH, DMF, 0°C to RT.
74%
HO OTBDPS
CO2EtTBDPSO 1
234
N3 OTBDPS
CO2EtTBDPSO
1)Tf2O, 2,6-lutidine,CH2Cl2, -78°C.
2) NaN3, DMF,0°C to RT, 12h.
88%
1
234
HCO2NH4, Pd/ C 10%, MeOH, RT, 2h.
98%H2N OTBDPS
CO2EtTBDPSO1
234
O OH
CO2EtO
O N3
O OTBDPS1
23
4LiAlH4, THF,
0°C
98%
O OH
O OH TBDPSCl, ImH, DMF, 0°C to RT.
80%
O OH
O OTBDPS1)Tf2O, 2,6-lutidine,
CH2Cl2, -78°C.
2) NaN3, DMF,0°C to RT, 12h.
90%
O N3
O OTBDPS
H2N OTBDPS
CO2EtTBDPSO1
234
NH2O
O OTBDPSPPh3, H2O, THF,
RT, 72h.
78%
N3HO
HO OTBDPSTFA, THF, H2O,
0°C, 2h.
55%N3O
OTBDPS
PPh3, DIAD,130°C,
0,1 mmHg
78%
Mitsunobu reaction
1) MeC(OMe)3, PPTS, CH2Cl22) AcBr, Et3N, CH2Cl2
3) K2CO3, MeOH.
87%
Sharpless epoxidation
PPh3, H2O, THF,RT, 72h.
87% NH2O
OTBDPS
SYNTHESIS OF AMINO-BUTANOL PRECURSORS
FORMATION OF NFORMATION OF N11-C-C22 LINKAGE BY PEPTIDE LINKAGE BY PEPTIDE COUPLINGCOUPLING
- FIRST STEP OF THE CAG STRATEGY -- FIRST STEP OF THE CAG STRATEGY -
34
P1 P2
Couplingreagent
Fmoc Bn
PyBOP78 %
HBTU49 %
Z tBu PyBOP80 %
Boc Bn PyBOP99 %
PEPTIDE COUPLING : SUBSTRATES AND PRODUCTSPEPTIDE COUPLING : SUBSTRATES AND PRODUCTS
"AMINO" PRECURSORS
NH2
CO2EtTBDPSO
TBDPSO
NH2
O
TBDPSO
O
NH2
O
TBDPSO
amino-ester
amino-acetonide
amino-epoxide
OP2HO2C
P1HN
L-SERINE
NH
NHFmocCO2Et
OOBn
TBDPSO
TBDPSO
PEPTIDECOUPLING
100 %
NH
NHFmoc
OOBn
O
TBDPSO
O
100 %
NH
NHP1
OOP2
O
TBDPSO
23
12'
1'
3'
4'
a : PyBOP, DIEA, CH2Cl2
b : HBTU, DIEA, DMF
a
a
a or b
* 25 % of epimerization in C2 position
*
COUPLINGPRODUCTS
YIELD
35
1,4-diazepan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
NH2
PO
amino-butanol
PO
Y
NH
H2N
O
PO
OH
1
2
34
5
6 7
Y
PO
N-alkylation
peptidecoupling
N-alkylationNH2
CO2H
NHPO
PO
OH
12
34
5
6
7
peptidecoupling
H2N
O
OH
HO
L-serine
CAG STRATEGY
ACG STRATEGY
ACCESS TO THE SCAFFOLD BY DIRECT COUPLINGACCESS TO THE SCAFFOLD BY DIRECT COUPLING
36
1,4-diazepan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
NH2
PO
amino-butanol
PO
Y
NH
H2N
O
PO
OH
1
2
34
5
6 7
Y
PO
N-alkylation
peptidecoupling
N-alkylationNH2
CO2H
NHPO
PO
OH
12
34
5
6
7
peptidecoupling
H2N
O
OH
HO
L-sérine
CAG STRATEGY
ACG STRATEGY
INTRAMOLECULAR PATHWAY
INTERMOLECULAR PATHWAY
ACCESS TO THE SCAFFOLD BY DIRECT COUPLINGACCESS TO THE SCAFFOLD BY DIRECT COUPLING
37
NH
H2N
OPOPO
R
R= O, O-(SO2)-O
NHPPO
RH2N
OPO
PO
NH
H2N
OPOPO
O
NHPPO
OH2N
OPO
PO
FORMATION OF NFORMATION OF N44-C-C55 LINKAGE BY LINKAGE BY NN-ALKYLATION-ALKYLATION
H2N
OPO
PONHP
PO
PO
Y
NH
H2N
OPO
PO
PO
Y
Y= Br, OTs
NH
NH2CHO
OPO
PO
PO
NHP
CHOPO
PO
NH2
OPO
PO
Reductive amination
INTRAMOLECULAR PATHWAY (CAG Strategy)
INTERMOLECULAR PATHWAY (ACG Strategy)
38
Cl3C-C(NH)-OtBu,cyclohexane, CH2Cl2,
50°C, 3h.
100 %
DBU, THF, RT, 2h.
100 %
BnOCO2H
NHFmoc
BnOCO2tBu
NHFmoc
BnOCO2tBu
NH2
PP11 PP22 Opening conditionsOpening conditions YieldYield
HH H.HClH.HCl
ttBuOH, NaH, 100°CBuOH, NaH, 100°C
CsCs22COCO33, DMF, 65°C, DMF, 65°C
CsCs22COCO33, DMF, 110°C, DMF, 110°C--
BnBn TBDMSTBDMS
ttBuOH, NaH, 100°CBuOH, NaH, 100°C
CsCs22COCO33, DMF, 110°C, DMF, 110°C
MeOH, EtMeOH, Et33N, 60°CN, 60°C
Yb(OTf)Yb(OTf)33, (Et, (Et33N), DCM, TAN), DCM, TA
--
ttBuBu BnBn Yb(OTf)Yb(OTf)33, DCM, RT, 7 days, DCM, RT, 7 days 65 %65 %
N3CO2P1
NHHO
OP2
TBDPSO
N3CO2tBu
NHHO
OBn
TBDPSON3
TBDPSO
O
tBuO2C
H2N
OBn
NH
H2N
OPOPO
O
NHPPO
OH2N
OPO
PON3
TBDPSO
OH2N
OP2
OP1O
FORMATION OF NFORMATION OF N44-C-C55 LINKAGE BY LINKAGE BY NN-ALKYLATION :-ALKYLATION : NUCLEOPHILIC ATTACK OF AN ACTIVATED PRIMARY NUCLEOPHILIC ATTACK OF AN ACTIVATED PRIMARY
CARBONCARBON INTERMOLECULAR PATHWAY
BnOCO2H
NHBoc
BnOCO2H
NH3Cl
HCl 3M/AcOEt, RT, 1h.
100 %
HOCO2Bn
NH3Cl
TBDMSOCO2Bn
NH2
TBDMS-Cl, ImH, DMF,0°C to RT, 15h.
70%
TBDMSOCO2Bn
NHCHO
25%
+
PP11 PP22
HH H.HClH.HCl
BnBn TBDMSTBDMS
ttBuBu BnBn
39
NH
NHFmoc
OBnOTBDPSO
O piperidine,DMF, RT
65% NH
NH
OBn
OTBDPSO
HO
INTRAMOLECULAR PATHWAY
deprotection
XNH
H2N
OBnOTBDPSO
O
Acid conditions :Yb(OTf)3, (Et3N), CH2Cl2, RT, 6 days.LiNTf2, CH2Cl2, RT, 48h.
Basic conditions :Cs2CO3, DMF, RT to 110°C, 20h.tBuOH, NaH, 100°C.
« Neutral » conditions :MeOH, RT, 15 days.iPrOH, RT, 18h.iPrOH, 50°C, 4 days.
X
epoxide opening
FORMATION OF NFORMATION OF N44-C-C55 LINKAGE BY LINKAGE BY NN-ALKYLATION :-ALKYLATION : NUCLEOPHILIC ATTACK OF AN ACTIVATED PRIMARY NUCLEOPHILIC ATTACK OF AN ACTIVATED PRIMARY
CARBONCARBON
40
« -stacking »interactions
O
Si
O
HN
OO
NH2H
Primary carbon atomof epoxide ring
Amine functioninvolved in
epoxide ring opening
MOLECULAR MODELING OF AMINO-EPOXIDEMOLECULAR MODELING OF AMINO-EPOXIDE
« -stacking » interactions
41
O CO2P
NHPO
O
OP
N3
CHOPO
PO
CO2P
H2N
OP
L-serineazido-aldehyde
1
3
2
4
1
23
4N3
CHOPO
POazido-aldehyde
1
23
4
OTBDPSO
OLiBH4, MeOH,Et2O, 0°C, 4h.
83%
HO
NH
NH2CHO
OPO
PO
PO
NHP
CHOPO
PO
NH2
OPO
PO
Reductive amination
NH2CO2P
NHPO
PO
OP21
34
NN-ALKYLATION BY REDUCTIVE AMINATION-ALKYLATION BY REDUCTIVE AMINATION
NH
NHPO
PO
OP
O
target diazepanone
O
CHOPO
O4
3
21
acetonide-aldehyde
CO2P
H2N
OP
L-serine
reductive amination
reductive amination
functionalizationof the diol moiety
functionalization of the diol moiety
peptidecouplingCO2Et
OTBDPSO
O(ClCO)2, DMSO, Et3N,
CH2Cl2, -78°C, 2h.
93%
CO2Et
OTBDPSN3
OTBDPS DIBAL-H (1M in toluene),CH2Cl2, -78°C, 2h.
96%
SYNTHESIS OF PRECURSORS INVOLVED IN REDUCTIVE AMINATION
1/ Aldehyde derivatives
P= TBDPS
O
CHOPO
O4
3
21
acetonide-aldehyde
42
YYARAR
8484
5858
2727
4747
YYARAR
8484
5858
2727
4747
YYZClZCl
8484
9595
YYTFATFA
7676
8383
O CO2R1
NRTBDPSO
OBn
O
O
NRTBDPSO
OBn
O
CO2R1
FUNCTIONALIZATION OF THE DIOL FUNCTIONALIZATION OF THE DIOL MOIETYMOIETY
Cl3C-C(NH)-OtBu,cyclohexane, CH2Cl2,
50°C, 3h.
100 %
BnOCO2tBu
NHFmoc
H2N
OBn
L-serine
R1O2C
N3
CHOTBDPSO
TBDPSO azido-aldehyde
1
23
4
NN-ALKYLATION BY REDUCTIVE AMINATION-ALKYLATION BY REDUCTIVE AMINATION
TFA, CH2Cl2,RT, 30 min.
100 %
TFA, CH2Cl2,RT, 30 min.
100 %
BnBr, K2CO3,DMF, RT, 3h
100 %
BnOCO2Bn
NHBoc
EtI, Cs2CO3,CH3CN,
reflux, 1h30
100 %
BnOCO2Et
NHBoc
Aldehyde Aldehyde derivativederivative
RR11
BnBn
ttBuBu
ttBuBu
EtEt
O OP
CHOOBnO
CO2Bn
NH2.CF3CO2H
BnOCO2Et
NH2.CF3CO2H
BnOCO2tBu
NH2
reductive amination
BnOCO2H
NHBoc
2/ Serinyl derivatives
N3CO2R1
NHTBDPSO
TBDPSO
OBn21
34
O
CHOTBDPSO
O4
3
21
acetonide-aldehyde
H2N
OBn
L-serineR1O2C
reductive amination
R = H
R = Z
ZCl, K2CO3, DMF, TA, 1h.
OH CO2R1
NZTBDPSO
OBn
R'O
TFA, H2O, THF,
0°C, 3h.
R’ = H
R’ = TBDPS
TBDPSCl, ImH, DMF,
0°C to RT, 15h.
1/ Tf2O, 2,6-lutidine, CH2Cl2, -78°C, 2h.
2/ NaN3, DMF, 0°C to RT, 15h.
N3 OP
CHOPO
YYN3N3
8585
--
1/ Step 12/ NaBH3CN, EtOH abs., 18 h.
1/ Step 12/ NaBH3CN, EtOH abs., 18 h.
BnOCO2H
NHFmoc
DBU, THF, RT, 2h.
100 %
SYNTHESIS OF PRECURSORS INVOLVED IN REDUCTIVE AMINATION
Step 1 : reductiveStep 1 : reductiveaminationamination
DIEA, DCM, 4Ả molecular DIEA, DCM, 4Ả molecular sieves, RT, 15h.sieves, RT, 15h.
Ti(OTi(OiiPr)Pr)44, DCM, RT, 3h, DCM, RT, 3h
Ti(OTi(OiiPr)Pr)44, DCM, RT, 3h, DCM, RT, 3h
YYTBDPSTBDPS
9494
9090
43
1,4-diazepan-2-one
NH
NH
O
PO
POC B
OH1 2
3
45
6
7
NH2
PO
amino-butanol
PO
Y
NH
H2N
O
PO
OH
1
2
34
5
6 7
Y
PO
N-alkylation
peptidecoupling
N-alkylationNH2
CO2H
NHPO
PO
OH
12
34
5
6
7
peptidecoupling
H2N
O
OH
HO
L-serine
CAG STRATEGY
ACG STRATEGY
ACCESS TO THE SCAFFOLD BY DIRECT COUPLINGACCESS TO THE SCAFFOLD BY DIRECT COUPLING
44
NH2CO2H
NR2R3O
OBn
TBDPSO
N3CO2R1
NR2R3O
OBn
TBDPSO
N3CO2R1
NR2R3O
OBn
TBDPSO
NH2CO2R1
NR2R3O
OBn
TBDPSO
NH2CO2H
NHR3O
OBn
TBDPSO
NH
NHR3O
OBn
TBDPSO O
Target diazepanone
NH2CO2R1
NR2R3O
OBn
TBDPSON CO2R1
NR2R3O
OBn
TBDPSO Bu3P
RR11 RR22 RR33 Azido reductionAzido reduction YielYieldd
DeprotectionDeprotection YielYieldd
PeptidePeptide
CouplingCouplingYielYiel
dd
BnBn ZZ TBDPSTBDPS HH22, Pd/C 10 %, MeOH, AcOEt, RT, 24h., Pd/C 10 %, MeOH, AcOEt, RT, 24h. DCC, HOBt, DCM, RTDCC, HOBt, DCM, RT --
NH
NHR3O
OBn
TBDPSO O
Target diazepanone
NH2CO2H
NR2R3O
OBn
TBDPSO
EtEt HH TBDPSTBDPS 1 1 nnBuBu33P, toluene, RT (3h) to reflux (5h)P, toluene, RT (3h) to reflux (5h) 2 2 TFA, THF, HTFA, THF, H22O, RT, 15h.O, RT, 15h. --
NH
NHR3O
OBn
TBDPSO O
azido reduction
FORMATION OF NFORMATION OF N11-C-C22 LINKAGE BY PEPTIDE COUPLING LINKAGE BY PEPTIDE COUPLING
deprotection
peptide coupling
deprotectionAND
1 2
X
X
RR11 RR22 RR33 FormationFormation
BnBn ZZ TBDPSTBDPS Reductive aminationReductive amination
EtEt HH TBDPSTBDPS Reductive aminationReductive amination
ttBuBu HH HH Epoxide ring openingEpoxide ring opening
ttBuBu HH TBDPSTBDPS Reductive aminationReductive amination
R2 = H
ttBuBu HH HH
HCOHCO22NHNH44, Pd/C, MeOH, , Pd/C, MeOH,
RT, 20 minRT, 20 min
5151
TFA, DCM, RT, 20h.TFA, DCM, RT, 20h.
100100
DCC/ HOBt, DIEA, DCC/ HOBt, DIEA, DCM/ DMF, RT, 24h.DCM/ DMF, RT, 24h. PyBOP, DIEA, DCM, PyBOP, DIEA, DCM, RT, 24hRT, 24h
--
ttBuBu HH TBDPSTBDPS 7171 100100
EDCI/ HOBt, DIEA, EDCI/ HOBt, DIEA, DCM/ DMF, RT, 24h.DCM/ DMF, RT, 24h. DCC, DIEA, DCM, RT.DCC, DIEA, DCM, RT.
--
45
NH
O
O
HO
H2N
O
SiO
Si
H
« hydrophobic site »
-stacking interaction
-stacking interaction
acid function involved inpeptide coupling
amine function involvedin peptide coupling
O O
H
NH
O HO
NH2
O
Si
H hydrophobicinteractions
MOLECULAR MODELING OF THEMOLECULAR MODELING OF THE « COMPLEX AMINO-ACIDS » « COMPLEX AMINO-ACIDS »
bis-O-silylated compound
Mono-O-silylated compound
-stackinginteraction
46
OP
NH2CO2H
NPPO
PO
OP
NR
NPPO
PO
O
OP
NH
NHP
O
R
PO
O
OHHO
O
NH2
NH
N
O
R2
O
O
O
OHHO
O
NH2
NH
N
O
R2
O
O
O
OHHO
O
NH2
NH
N
O
R2
O
O
O
OHHO
O
NH2
NH
N
O
R2
O
OX
O
OPPO
O
N3
PO
PHN
O
O-GLYCOSYLATION
PEPTIDE COUPLING
N-ALKYLATION
⇒ RING CLOSURE ?
CONCLUSION AND PERSPECTIVESCONCLUSION AND PERSPECTIVES
1
5
2
34
6
7
⇒ HOAt
47
NH2CO2H
NHHO
OBn
PO
NH
NHHO
OP
POO
NH
NHHO
O
POO
O
OO
NHP
NH
NR1R2O
O
R3OO
O
HOOH
NH2
N3
PO
OH2N
OBn
OtBuON3CO2tBu
NHHO
OBn
PO
F
O
OO
NHP
R1, R2, R3 =O
OHAHO
OH
NHN
O
O
A = OH, NH2, NHAc...
(CH2)n CH3
(CH2)n CO2H
TOWARDS A NEW FAMILY OF POTENTIAL TOWARDS A NEW FAMILY OF POTENTIAL ANTIBIOTICSANTIBIOTICS
N-Alkylation of L-serine tert-butyl ester
+ Intramolecular peptide Coupling
+ O-Glycosylation of diazepanone heterocycle
Ribosyl-diazepanone scaffold
+ R1/ R2/ R3
Family of powerful MraY inhibitors
⇒ Biologic evaluation (Laboratoire des Enveloppes Bactériennes etAntibiotiques – Dr D. Blanot – Dr. D. Mengin-Lecreulx)