structure & modifications

Vitamin B12

Mikoaj Chromiski Group XV Institute of Organic Chemistry PAS

 Introduction Structure Nomenclature (not only) according to IUPAC Possible modifications Modifications of carboxyl groups Modifications on C-10 Modificatons on Ribose-O 5 Functionalisation of Co atom

General information
water soluable Key role in the normal functioning of the brain and nervous system anf formation of blood Affects DNA synthesis and regulations 2mg/day for adult concentration in blood : 165-680 ng/l Sources in nutrition: meat (liver), fish, eggs, milk

Why it is interesting for chemists?

Drug design, drug delivery Electrochemistry Catalysis

Enzymology

Some History
Discovery : 1948 by Folkers at Merk Structure: very looooong process First total synthesis accomplished 1973 Woodward Eshenmoser

H2 2NOC

CONH2 2

H2 2NOC H2 2NOC

N N HN N CN N NH Co Co
N HN N NH N N

CONH2 2

Structure
O N

CONH2 2

NH

HO O
P O

-O O

HO
Molecular Weight Molecular Formula 1355.36[g/mol] C63H88CoN14O14P

IUPAC Name: cobalt(3+);[(2R,3S,4R)-5-(5,6-dimethylbenzimidazol-1-yl)4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2R)-1-[3-[(1R,2R,3R,5Z,7S,10Z,12S,13S,15Z,17S,18S,19R)2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)3,5,8,8,13,15,18,19-octamethyl-2,7,12,17-tetrahydro-1H-corrin-24-id-3-yl] propanoylamino]propan-2-yl] phosphate; cyanide cyanocobalamin CN-Cbl Cyanocob(III)alamin Vitamin- B12 V2876_SIAL a (5,6Dimethylbenzimidazolyl)cyanocobamine

Nomenclature
H2NOC H2NOC H2NOC N O NH

CONH2

N CN N Co N

CONH2

CONH2 N N

HO O
P -O O O HO

5 3 2 4

7 6

AN
1 19 21 24

N N
14

B
9 11

22 10

18

D
17

NH
16 15

23

C
13

12

b
H2NOC

c
CONH2

a
H2NOC

H2NOC

N CN N Co N N

CONH2

CONH2

O NH

N N

HO O
P O

-O O

HO

ribose 5'

b
L1

N Co N N N
L2

H2NOC

CONH2

(ligand in Cob) cobalamine if L = CN :

CONH2

H2NOC H2NOC

N L N Co N N

cyanocobalamine (vit. B12) or CN-Cbl if L = H2O: aquacobalamine (vit. B12 a) or (aq)Cbl

CONH2 O NH N N

HO O
P -O O O HO

H2NOC

CONH2

H2NOC H2NOC

NL N Co N N

CONH2

Cobyrinic acid (Cby) for example

CONH2

HOOC

(CN)MeCby Me(aq)Cby

H2NOC

CONH2

H2NOC H2NOC

Cobinic acid (Cbi)

NL N Co N N CONH2

for example (CN)MeCbi (CN)(2-OAcBu)Cbi

CONH2
O NH RO

for more complex ligands, we use specific abbreviations ( for example aminoacids abbreviations)

Some simplifications
H2NOC CONH2 H2NOC H2NOC

N L N Co N N

CONH2

L
L

=
CONH2

or
Co

Co N OH

O NH

N N

HO O
P -O O O HO

hydrolysis/conjugation

H2NOC

CONH2
hydrolysis/conjugation

H2NOC H2NOC

N CN N Co N N

CONH2

Modifications
CONH2 O NH N N

hydrolysis/conjugation

HO O
P -O O O HO

lactonisation/reduction/conjugation

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

CONH2

CONH2 O NH N N

HO O
P -O O O HO

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

nitration/reducrion/conjugation

CONH2

CONH2 O NH N N

HO O
P -O O O HO

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

CONH2

CONH2 O NH N N

HO O
P -O O O HO

O
direct conjugation

conjugation on cobalt

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

CONH2

CONH2 O NH N N

HO O
P -O O O HO

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

CONH2

CONH2
hydrolysis/conjugation

O NH

N N

HO O
P -O O O HO

Modification of carboxylic groups

H2NOC CONH2 H2NOC H2NOC N CN N Co N N ROOC CONH2 COOR

ROH/H2SO4
CONH2

ROOC ROOC N CN N Co N CN N COOR

KCN

O NH

N N

COOR O

HO O
P -O O O HO

ROOC

Murakami, Hisaeda et al. Bull.Chem. Soc. Jpn. 1983, 56, 3642 Murakami, Hisaeda et al. Bull.Chem. Soc. Jpn. 1984, 57, 2091 Murakami, Hisaeda et al. Bull.Chem. Soc. Jpn. 1984, 57, 405

(CN)2Cob(III)C1ester (CN)2Cob(III)C2ester (CN)2Cob(III)C3ester (CN)2Cob(III)C4ester Yields up to 85%

ROOC

COOR

ROOC

COOR

ROOC ROOC N CN N Co N CN N COOR

HClO4 aq

ROOC N ROOC N

CN N Co N H2O

COOR

ClO4-

COOR COOR

ROOC ROOC

ROOC

COOR

ROOC

NaBH4
ROOC

N Co N

N COOR N

ClO4-

COOR

ROOC

[Cob(II)ester] ClO4

H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

CONH2

0,4M HCl
CONH2 O NH N N

HO O
P -O O O HO

Russel-Jones et al. Bioconjugate Chem. 1995, vol 6, 34 Anton et al. J. Am. Chem. Soc., 1980, 120, 2215 Russel-Jones et al. Bioconjugate Chem. 1996, vol 7, 217

HOOC

CONH2

H2NOC

H2NOC CONH2 H2NOC N CN N Co N N COOH H2NOC

CONH2

H2NOC H2NOC

N CN N Co N N

H2NOC CONH2 H2NOC

N CN N Co N N

CONH2

CONH2 O NH HO O P -O O O HO N N O O NH HO O P -O O O HO N N O

CONH2 O NH HO O P -O O O HO N N O

COOH

17%

16%

6%

H2NOC H2NOC H2NOC

CONH2 N CNN Co N N

H2NOC H2NOC

CONH2 N CNN Co N N

CONH2

H2N EDAC

H2NOC

CONH2

COOH O NH N

+
pH 6.d buffer NH2
O NH N N HO O O P O -O O HO CONH

HO O O P O -O O HO

H2N

65%

Russel-Jones et al. Bioconjugate Chem. 1995, vol 6, 34

O
H2NOC H2NOC H2NOC CONH2 N CNN Co N N H2NOC H2NOC CONH2 H2NOC

O
N CNN Co N N CONH2

NBS, AcOHaq
CONH2

CONH2 O NH N N HO O O P O -O O HO

O NH

HO O P O
-O O

N O HO

100%

Keese et al. Helvetica Chimica Acta, 1998, vol.81, 1105 Russel-Jones et al. Bioconjugate Chem. 1996, vol 7, 217

O O
N

B
CONH2

H2N

NH2

reflux O N H 70% NH2

OH
N

B
CONH2

Russel-Jones et al. Bioconjugate Chem. 1996, vol 7, 217

O
H2NOC H2NOC H2NOC

O
N CNN Co N N

O
MeOOC CONH2 MeOOC MeOOC

O
N CNN Co N CNN COOMe

MeOH/H2SO4
CONH2 O NH N

HO O O P O -O O HO

COOMe

MeOOC

70%

Keese et al. Helvetica Chimica Acta, 1998, vol.81, 1105

MeOOC MeOOC MeOOC

OH

O
MeOOC

NaBH4

N CNN Co N CNN

COOMe

COOMe

MeOOC MeOOC

O
N CNN Co N CNN COOMe

MeOOC

O
COOMe MeOOC

OH Zn/AcOH
MeOOC MeOOC N CNN Co N CNN

MeOOC

COOMe

80%
COOMe

MeOOC
Murakami, Hisaeda et al. Chemistry Letters, 1992, 909 Keese et al. Helvetica Chimica Acta, 1998, vol.81, 1105

O OH
N

O 1. Cl O

O CCl3
N

B
COOMe

N H B
COOMe

NH2

H2N

NHBoc

Et3N, DCM 2. CF3COOH

33% COOH HOOC N N 56% O N H

N
COOH

COOH

COOH

N O O O

O NH

B
COOMe

Scheffold et al. Helvetica Chimica Acta, 1997, vol 80, 1183

O O OH
N

O B

O O EDC, DMAP O O
COOMe N

B
COOMe

+
OH

O O 65%

Keese et al. Helvetica Chimica Acta, 1998, vol.81, 1105

Si(OMe)3
MeOOC COOH

+
MeOOC

HN

MeOOC N MeOOC N

Si(OMe)3
CN N Co H2O N COOMe

ClO4-

H2N DEPC

MeOOC N MeOOC N

CN N Co CN N

COOMe

COOMe COOMe

MeOOC MeOOC

O O O

P
CN

MeOOC

OH 1.
N CNN Co N CNN

COCl OMe

MeOOC MeOOC

MeO O C O

MeOOC MeOOC

COOMe

2. NaBH4
COOMe

MeOOC

N N Co N N

COOMe

68%
COOMe

MeOOC

MeOOC

Murakami, Hisaeda et al. Chemistry Letters, 1992, 909

Nitration at C - 10
MeOOC COOMe MeOOC COOMe

MeOOC MeOOC N CN N Co N CN N

COOMe

1. NO2BF3 2. NaBH4

MeOOC MeOOC N CN N Co N CN N

COOMe

NH2

COOMe

COOMe

MeOOC

MeOOC

13%

Murakami et al., J. Chem. Soc. Perkin Trans 2, 1995, 1175

MeOOC

COOMe

O O
MeOOC MeOOC N CN N Co N CN N COOMe

COOMe

O O O
N N

COOMe

OHC

B O N O C

NH2

O O O O
COOMe

COOMe

MeOOC

56%

Hisaeda et al. Tetrahedron Letters, 2003, 44, 6421

Combibation of lactone and C-10 approach

O
MeOOC

MeOOC MeOOC N CN N Co N CN N

COOMe

H N O
COOMe

MeOOC
Murakami et al., J. Chem. Soc. Perkin Trans 2, 1995, 1175

5 O Ribose modifications
H2NOC CONH2 H2NOC CONH2 H2NOC H2NOC N CN N Co N N H2NOC CONH2 H2NOC N CN N Co N N

CONH2

CDT

CONH2 O NH N N O NH O N N

CONH2

HO O
P -O O O HO

HO O
P -O O O

O O

O
McEwan et al. Bioconjugate Chemistry, 1999, vol. 10, 1131

N N

CN
H2NOC CONH2

GlyOMe

Co N 95% O O N H OH O

H2NOC H2NOC

N CN N Co N N

CONH2

CONH2 O NH N N

CN Co diamine N O O N H 65-98% NH2 n

HO O
P -O O O

O O

N N

Thanks for attention!