synthesis of vitamins & cofactors - uni konstanz · 2019-10-07 · thiamine synthesis of...
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17.07.2014, Darius Schwazrer
Synthesis of Vitamins & CofactorsGaich Group Seminar
Definition
Cofactor/Coenzyme: A nonprotein component of enzymes is called the cofactor. If the cofactor is organic, then it is called a coenzyme. Coenzymes are relatively small molecules compared to the protein part of the enzyme. Many of the coenzymes are derived from vitamins
Protein + Cofactor/Coenzyme = Holoenzyme Proteinpart of Enzyme = Apoenzyme Vitamin:
1912 Kazimierz Funk defined that the essential substances contain an amine function are called Vita = life; amine
!Essential, organic compounds which are not synthesized in the human or animal organism or formed only in insufficient amounts . Must be regularly consumed with diet
Vitamin A & ß-Carotene
Retinol acetate (BASF)
3
ß-Carotoene (BASF)
Me
MeMe
O
Me
MeMe Me Me
OAc
Retinoacetate (4)
ClMg
Me
MeMe OH
Me
Me
MeMe Me
PPh3
HSO4
1. THF
98 %
2. PPh3, H2SO4, i-PrOH, Heptane
95 %
Me
AcOO
3. BuLi, THF, - 50 °C
80 %1 2 3
c. KOAc, MeOH, rflx. then KOH, H2O, (90 %)d. Acetone, MnO2 (85 %)e. EtOAc, Lindlar, H2 (95 %)f. Benzene, Iodine, hexane, hv (?? %)
BrMgC CMgBrO
a. Et2Othen 10% H2SO4
52 %
b. PBr3, (CH2OH)2
75 %
OHHO
CH2Br
BrH2C
85 6 7
Me
MeMe Me
PPh3
OO
2x
DMF, NH3, thenH2O2 isomerization
Me
MeMe Me Me
Me Me
Me
Me Meβ-Carotin (9)
80 %
8
3
Mildner; Weedon Journal of the Chemical Society, 1953 , 3294, Inhoffen; v.der Bey Justus Liebigs Annalen der Chemie, 1953 , 583.
Ascorbic Acid
4
Vitamin C synthesis by Reichstein process
step 3+4 can be replaced by oxidation with Air & PtO
Reichstein synthesis was optimized so that each step gives over 90 % yield !Overall yield from D-glucose is now 60 % !Direct synthesis via microalgae possible, but yield and concentration are too low
OOH
OH
OH
OHHO
D-glucose (10)
O
OHOO
Me
Me
O O
Me Me
OHO
OH
OH
OHHOOH
OH
OH
OH
OHHO 37 %
3. Acetone,H2SO4
AcetobactersuboxydanspH 4-6, 30-35 °CH2, RaNi
quant. >90 %
11 12 13
OH
OHOHO
O
HO
OHO
OH
OH
OHHO
O
4.NaOCl, NiCl, > 93 %5. H2O, H+, 82 %
14
6. HCl, PhMe, Δ
> 60 %
Ascorbic acid (15)
T. Reichstein, A. Güssler Helv. Chim. Acta 1934, 17, 311; Bullmanns Enzykl. Tech. Chem., 3. Ed., Vol 18, 233 ff.
Ascorbic Acid
Metabolism of Vitamin C
5
Industrial use • Inhibition of nitrosamine in
cured meat• Improvement of flour quality• Increasing of clarity of wine &
beer• protecting fruits from browning
Economic aspects • production >
600 kt/a• cost ~3000 €/t
Biological activity • anti oxidant• radical scavenger• electron donator for 8
enzymes(hydroxylases)
• prevents from scurvy
HO
OHHO O
O
OH O
OHHO O
O
OHO
OHHO O
O
OHO
OHHO O
O
O
OH
HOO
OOHO
OH
-H+
+H+
-e- -H+
+H+
pka = -0.86pka = 4.1
-e-
O
OHHO O
O
O
+H2O
-H2O
+H2O
-H2O HO
OHHO O
O
OHOHOH
15
16 17 18
Thiamine
Synthesis of Vitamin B1 Step 1. Synthesis of 22
6
22 and 23 commercially available
Step 2. Condensation & formation of thiazolium ring
Me
N
NH2
CN CN
Me
N
NH
CN
CN
a. i-PrOH, max. 40 °C
87 %
b.Na(powdered), PhEt, Styrol & PhNHMe in THF over 2 h, rt, then rflx, 20, then H2O
77 %
N
NH
Me
NH2
CNc. MeOH, NH3, RaCo,
110 °C 60 atm, 13 h
50 %
N
NMe
NH2
NH2
19 20 21 22
N
NMe
NH2
NH2
O
Cl
AcO
O
SH
AcO1. KSH, MeOH,
0 °C
74 %
N
NMe
NHN2. 80 °CMe2N
OMe
OMe
89 %
3. HCO2H, < 40 °C
89 % OHS
N+
Me
NH2NMe
NCl-
Thiamine (26)
2324
22 25
A. Edenhofer, H. Spielberg, W. E. Oberhänsli Helv. Chim. Acta 1975, 58, 1230; P. Contant, L. Forzy, U. Hengartner, G. Moine Helv. Chim. Acta 1990, 73, 1300.
Thiamine
Biological activity • Thiaminediphosphate (TDP) is the active form• Essential for carbohydrate & energy metabolism
• Present in pyruvat dehydrogenase, oxoglutarate dehydrogenase,transketolase, pyruvate decarboxylase, 2-hydroxyphytanoyl-CoA lyase.
7
Deficiency • Beriberi —> effects peripheral nervous system; cardiovascular system,• Polyneuropathy• Anemia• many more
N
S
RMe
PPO
HN
S
RMe
PPO
N
S
RMe
PPO
MeHO
OOH
N
S
RMe
PPO
OH
Me
CoAS Ph
O
N
S
RMe
PPO
OH
Me
O
Ph
MeO
PhO
Me
OHO
O
27 29
Biotin
Vitamin B7 Synthesis by Hoffmann-La Roche
8
US 2 489 232 (Roche; 1949; appl. 1946); US 2 489 233 (Roche; 1949; appl. 1946), US 2 489 234 (Roche; 1949; appl. 1946), US 2 489 235 (Roche; 1949; appl. 1946), US 2 489 236 (Roche; 1949; appl. 1946), US 2 489 237 (Roche; 1949; appl. 1946), US 2 489 238 (Roche; 1949; appl. 1946), US 2 519 720 (Roche; 1950; appl. 1948), US 3 740 416 (Roche; 1973; CH-prior. 29.11.1969).
HO2C
CO2H BnN NBn
O
HO2C CO2H
BnN NBn
O
HO2C CO2R
BnN NBn
O
RO2C CO2H
BnN NBn
S O
O1. Br2, H2O, 90 %2. BnNH2, MeOH,
90 %3. COCl2, KOH
5. LiBH4, THF6. AcSK, DMF,
150 °C
> 90 %
4. PhH, C6H12O, Δ, then(+)-Ephidrine, iPrOH,80 °C
2 steps
30 3132
33
34
OH
NHO
HHN
H S
OBnN NBn
S
O
OMe
BnN NBn
S
O
Br
7. then AcOH, Δ
8. Ra/Ni, H2 40 atm.35 °C, MeOH
9. 48% HBr,95 °C, 2 h
82 % 90 %
10. Malonic acid diethyl ester,NaOMe, PhMe, 80 °Cthen, 48% HBr, rflx.
(25% over all)
35 36
Biotin (37)
ClMg OMe
Biotin
Biological activity • present in carboxylases (pyruvat-, acetyl-CoA-, propionyl-CoA-, methylcrotonyl-
CoA-carboxylase)• catalyzes carboxylation as a CO2 carrier• Formation of malonyl-CoA from acetyl-CoA
9
Biotin deficiency causes Neurological symptoms (depression, lethargy, etc.)
ATP HCO3- HN NH
S
O
CO2H4
+ ADP + OP
HO
O
OHOH
O
N NH
S
O
CO2H4
HO
O
CoAS
OO
SCoA
O
HO
Biotin+
37
3839
Cobalamine
Vitamin B12 • Synthesis by fermentation• No industrial total synthesis available
• Total synthesis with 113 steps, 12 PhD, 91 Postdocs,17 years
• optimized to 70 steps• cobalt can be complexed by CN-, SO32-, OH-, halides • 10t/a; market volume 77*106 EUR!Biological activity • catalyzes 1,2 H shift• Coenzyme for Methionin-synthase & Methylmalonyl-CoA-mutase• Hydroxy-cobalamine —> treatment of cyanide poisoning• deficiency causes anaemia
10
Crowfoot-Hodgkin, D. et al. Nature, 1956, 178, 64; Woodward, R.B. Pure & Appl. Chem. 1968, 17, 519; Woodward, R.B. Pure & Appl. Chem. 1971, 25, 283; Woodward, R.B. Pure & Appl. Chem. 1973, 33, 145; Eschenmoser, A.; Wintner, C.E. Science 1977, 196, 1410; Woodward, R.B. In Vitamin B12, Proceed. 3rd European Symposium on Vitamin B12 and Intrinsic Factor, Zagalak, B.; Friedrich, W., Eds., Walter de Gruyter: Berlin, 1979, P.37; Benfey, O. T.; Morris, P. Robert Burns Woodward: Architect and Artist in the World of Molecules. Philadelphia: Chemical Heritage Foundation, 2001.Nicolaou, K.C.; Sorenson, E.J.; Classics in Total Synthesis: Targets, Strategies, Methods. Weinheim, Germany: VCH, 1996.
N N
N N
Me
Me
Me
Me
H2NOC
H2NOC
H
H2NOCMe
MeMe
CONH2
Me CO2NH2
NHO
OMe PO
OO O
HO N
NMe
Me
CONH2
HO
CoCN
40
Calcitriol
Synthesis of Vitamin D3
11
MeMe
Me
HOH
H
H
OH1. Ac2O, Pyridine,
100 °C, 17 h
96 %
N
NO
O
MeMe
Br
Br
2. NaHCO3,Hexane,20 min.
0.5 Eq.
N
Me
MeMe
3. Xylene, Δ2 eq.
then pTSA, Dioxane, 70 °C, 30 min., 50 %
MeMe
OH
3
MeMe
Me
AcOH
H
H
OAc
MeMe
OAc
3 MeMe
Me
AcOH H
OAc
MeMe
OAc
3
41 4243
5. -15 °C KOH, MeOH, THF78 % Diacetate, 16 % Monoacetate
6. 450 W Hg, -5 °C, THF, hexane, 20 %25 % unconverted
7. KOH, MeOH, 30 h, quant.
MeMe
Me
HOH
OH
MeMe
OH
3
MeMe
H
MeMe
OH
8. Δ, Dioxane, 30 min
50 %
3
OHHO
Calcitriol (45)
44
US 3 993 675 (Hoffmann La Roche Inc.; 23.11.1976; prior 12.11.1973; 24.02.1975.
Calcitriol
Biological activity of Vitamin D3
• hormonally active vitamin• increases the Ca2+ concentration in the blood, by
increasing the uptake in the gastrointestinal tract• decreases the probability to get some of up to 17
different cancer diseases Hypervitaminose causes calcification of tissues and arteries !!
Production & prize • No t/a given• cost 50 $/kg
12
MeMe
H
MeMe
OH
3
OHHO
Calcitriol (45)
Folic Acid
Vitamin B9 synthesis
13
E. L. Bennett J. Am. Chem. Soc., 1952, 74 (9), pp 2420; S. Nishigaki, K. Ogiwara, F. Yoneda Chemical & Pharmaceutical Bulletin 1971, 19, 418, Wailer, C.W. et al.: J. Am. Chem. Soc. (JACSAT) 70, 19 (1948). Hultquist, M.E. et al.: J. Am. Chem. Soc. (JACSAT) 70, 23 (1948); Angier, R.B. et al.: J. Am. Chem. Soc. (JACSAT) 70, 25 (1948).; Boothe, J.H. et al.: J. Am. Chem. Soc. (JACSAT) 70, 27 (1948). US 2 436 073 (American Cyanamid; 1948; appl. 1945)., US 2 442 836 (American Cyanamid; 1948; appl. 1945).; US 2 443 078 (American Cyanamid; 1948; appl. 1945).; US 2 443 165 (American Cyanamid; 1948; appl. 1946).; US 2 444 002 (American Cyanamid; 1948; appl. 1946).; US 2 472 482 (American Cyanamid; -1949; appl. 1947).; US 2 477 426 (American Cyanamid; 1949; appl. 1948). US 2 547 501 (American Cyanamid; 1951; appl. 1946). US 2 599 526 (American Cyanamid; 1952; appl. 195 1).; US 2 719 157 (Shionogi; 1955; appl. 1951). US 2 956 057 (Kongo Kagaku Kabushiki Kaisha; 1960; J-prior. 1955).
Folic acid is metabolized to „THF“ what is the active compound
NH
H2N NH2
CN
CO2EtN
N NH2H2N
OH
N
N NH2H2N
OHH2N
2. H2SO4/H2O, NaNO2 5 °C,then room temperature
82%
3. MeOH, Pd/C, H21. MeOH, Δ, quant.
46 47 4849
NH
O
HO
OOH
OH2NCH2ClO
CHCl2
O OH
NH
O
NH
HN
O
N
H2N NH
NH
O
OH4. Dioxane, H2O,
NaHSO3, pH 4-55. AcOH, 2-mercapto- ethanol NaCNBH3,
85 %N
N NH2H2N
OHH2N
49 50 51 Tetrahydrofolic acid (52)
RNH
HN
O
N
H2N NH
NH
PLP, Serine
PLP, Glycine
Michael-Additionthen Retro-Mannich N
O
N
H2N NH
NH
HNR
NO
N
H2N NH
NH
NR
"THF" 5253 54
Folic Acid
Biological activity of „THF“
14
Economic aspects • 400 t/a available as pure
crystals• cost 36 - 77 €/kg• used for feed enrichment
Formyl transfer in the Histidine biosynthesis
Methyl transfer in the SAM biosynthesis
Deficiency • anaemia• neural tube defects• depression
Folic acid & derivatives (antifolates) are used in anti cancer therapy Acts as an antidepressant
NO
N
H2N NH
NH
NR
NADH + H+ NO
N
H2N NH
NH
NHRMe
NAD+
Homocysteine,Methioninesynthase "THF"
NH2
SO
OH
Methionine (56)5455
NADH
NAD+ NO
N
H2N NH
NH
NR
H2OHN
O
N
H2N NH
NH
NROHC
Histidine &
THF
NO
N
H2N NH
NH
NR
54 57 58
Nicotinamide
Vitamin B3 synthesis (Lonza)
15
active coenzyme
redox reactions using NAD+ & NADPH
CONH2
NO
OHOH
OPO
OO
PO
OO
ONN
OH
OX
H2N X=H NAD+ (63)X= PO3
2- NADP+ (64)
O
O
O
Me
Me Me
1. (NH4)3PO4, H2O,200 °C, 15 bar
68 %
NMe
Me
N
CO2H
2. HNO3, Δ6-8 Mpa
N
CONH2
3. NH3, > 200 °Cneat
96 % 85 %
Nicotinamide (62)Niacine (61)59 60
R
H2NOC
N-O
O
O-
OO
Malate (65)NAD+ (63)
H B-
H
Hmalate
dehydrogenase
R
H2NOC
N
NADH (66)
H H
-OO
O-
OO
Oxaloacetate (67)
R
H2NOC
N-O2C
O
ß-Aspartat-semialdehyde (68)
NADP+ (64)
HB
Hhomoserine
dehydrogenase
R
H2NOC
N
NADPH (64)
H H
NH3+
-O2C
OH
NH3+
Homoserine (69)
DOS 2 046 556 (Lonza; appl. 22.09.1970; CH-prior. 24.09.1969); US 2 905 688 (Abbott; 1959; appl. 1954); US 2 280 040 (SMA Corp.; 1942; appl. 1939); US 2 314 843 (American Cyanamid; 1943; appl. 1941); US 2 993 051 (Cowles Chem. Comp.; 18.07.1961; appl. 26.02.1958).
Nicotinamide
16
Economic aspects • world capacity >>22000
t/a• cost 10-100 $/kg• mainly produced in China
Vitamin B3 deficiency • Pellagra
diarrhea, dermatitis, dementia, death
• Mental confusion• Ataxia• Dilated
cardiomyopathy• peripheral neuritis• etc.
R
H2NOC
N
NADP+
H
R
H2NOC
N
NADPH
HR H
H3C O
HSH3C O
HR HS
H BBH
• re- (pro R) & si- (pro S) specific enzymes• stereospecific dehydrogenation also possible
Pyridoxine
Vitamin B6 synthesis
17
O
O-
NH3+
RO
O-
NH3+
RO
O-
NH3+
R
L-amino acid L-amino acid D-amino acid
PLP
Transamination
Decarboxylation Isomerization Retro-Aldol
1. EtOH, HCl, 35 °C,red. pressure
NH2
MeO
OH
80 %2. formamide,
slow to 105 °C,45 min
HN
MeO
OEt
CHO
3. P2O5, CHCl3,then KOH,MeOH, H2O
40 - 60 % N
O
Me
OEt80 %
O OMe Me
4.then HCl(aq)
180 °C
Me
NHOHO
OHPyridoxine (73)70 71 72
OH
OHOH
N Me NH
Me
OH
OOP
O
O
O
PyridoxalphosphatePLP (74)
Pyridoxine (73)
O
O-
NH3+
RO
O-
O
-OO
ketoglutarate (76)
O
O-
O
RO
O-
O
-ONH3
+
glutamate (78)
PLP
75 77
O
O-
NH3+
R
NH3+
RCO2
PLP
75 79
O
O-
NH3+
OH
R R
O O
O-
NH3+
PLP
80 81 82
Review: H. König, W. Böll Chem. -Ztg 1976, 100, 105; E. E. Harris et. al. J. Org. Chem. 1962, 27, 1962; DAS 1 470 022 (Merck & Co; appl. 10.05.1962; USA pripr. 15.05.1961, 16.01.1962); US 3 227 721 (Merck & Co; 04.01.1966; prior. 15.05.1961, 16.01.1962, 24.01.1965); US 3 227 724 (Merck & Co 04.01.1966; prior. 15.05.1962; 16.01.1962, 16.06.1964).
Pyridoxine
Mechanism for the decarboxylation reaction
18
NH
Me
O
NOiP H
(CH2)4EnzCO2-R2
NH2
NH
Me
O
NHOiP H
(CH2)4EnzHN
R2 CO2-
NH
Me
O
NOiP H
CO2-R2
Enz(CH2)4NH2
NH
Me
O
NOiP H
CO2-R2
NH
Me
O
NOiP H
R2
NH
Me
O
NOiP H
R2
NH
Me
O
NOiP H
(CH2)4Enz
R2 NH2
regeneration by transimination
H B B-
Economic aspects • world capacity
>>4100t/a• cost 50$/kg• mainly produced in
China
Pyridoxine deficiency causes • anaemia• nerve damage• skin problems
Used for treatment of Wilson’s & rheumatic diseases carpal tunnel syndrome, autism, schizophrenia, mental retardation, etc.
K. H. Blässler, Int. J. Vitam. Nutr. Res. 1988, 58, 105; A. H. Merill, J. M. Henderson Annu. Rev. Nutr. 1987, 7, 137.
Riboflavin
Vitamin B2 synthesis
19
An efficient & cheap synthesis of D-ribose (90) is essential for the industrial riboflavin production
Me
Me
NH2 OHOH
OH
OHO 1. MeOH, rflx., then
RaNi/H2 25 atm. Me
Me
HN OH
OH
OH
OH
70 %
83 8485
NO2
NN
Cl Me
Me
HN
Rib.
NN
PhNO2
2. EtOH, 10 °C
92 %
3. Barbituric acid, AcOH, rflx. 5 h
67 %
OH
Me
NNH
O
NOH
HOOH
N O
Me
Riboflavin (87)
86
OOH
OH
OH
OHHO
D-Glucose (10)
a. KOH, O2
OK
O
OH
OH
OHHO
b. CaCl2c. H2O, 140 °C
d. H2SO4f. Na/HgOCa0.5
O
OH
OH
OHHO
O
OH
OH
OHHO
8889 90
IJS 2 384 105 (Roche; 1945; appl. 1943).; Tishler, M. et al.: J. Am. Chem. Soc. (JACSAT) 69, 1487 (1947).; US 2 350 376 (Merck & Co.; 1944; appl. 1941).; US 2 370 093 (Merck & Co.; 1945; appl. 1941). similar processes: Tishler, M. et al.: J. Am. Chem. Soc. (JACSAT) 67, 2165 (1945).; US 2 807 61 1 (Merck & Co.; 1957; appl. 1955).
Riboflavin
Biological activity
20
Me
NNH
O
NR
N O
Me
Me
NH
NH
O
NR
HN O
MeSred Sox
Economic aspects • world capacity 2400t/a• cost 45-54 €/kg• produced by HLR,
BASF, Takeda, ADM
Deficiency • atrophic glossitis• seborrheic dermatitis• eye problems
(all in all nothingserious)
• Most reactions followa radical mechanism
• Is able to form Cl+species with oxygen &chloride
Me
NNH
O
NR
N O
Me
B HS-SH
R2
Me
NH
NH
O
NR
N O
Me
SSH R2B-
HB
Me
NH
NH
O
NR
HN O
Me
S SR2
Methoxatin (non vitamin)
Synthesis of Methoxatin • 1st. Synthesis by Corey in 50 mg scale
21P. Martin, E. Steiner, K. Auer, T, Winkler Helv. Chim. Acta 1993, 76, 1667.
NH2
OMe
NO2
NHCHOOMe
NH2
NHCHOOMe
N2BF4
NHCHOOMe
NN
Me CO2EtCOMe
1. Ac2O, HCO2H, 35 °C (92 %)2. Pd/C, H2, DMF, 35 °C, 2 bar (80 %)
3. EtOH, HCl, NaNO2,then HBF4, EtOH
4. Ethyl-2-methyl acetoacetate, NaOAc, EtOH
91 92 93 94
NHCHOOMe
HNN
Me CO2Et
NHCHOOMe
NHEtO2C
OMe
NHEtO2C
NH
CO2MeHO
CO2Me5. Ph 6.5-5.2 6. HCO2H, 80 °C
7. HCl, Acetone 80 °C (85 %)8. 2-oxoglutaconic acid-dimethylester, CH2Cl2 (73 %)
70 % 62 %
95 96 97
N
HN
HO2C
CO2H
OO
CO2H
9. Cu(OAc)2, CH2Cl2, HCl/Air (95 %)10. CAN, H2O, MeCN (69 %)11. LiOH, H2O, THF; then H2SO4 (73 %)
(13 % over all)
98
Synthesis of Vitamins & Cofactors
22
Thank you for your attention
Synthesis of Vitamins & Cofactors
23
General Literatur Ullmann's Encyclopedia of Industrial Chemistry, 2005 Wiley VCH Verlag GmbH & Co. KGaA, Weinheim (10.1002/14356007.a27_443) Pharmaceutical Substances: Syntheses, Patents, Applications, 4th Ed. 2001 Stuttgart, Thieme, ISBN-13: 978-3135584058. !Naturstoffe der chemischen Industrie, 1. Auflage 2007, Elsevier GmbH, München; ISBN-13: 978-3-8274-1614-8. !Handbook of Vitamins, 4th Ed. 2007, Taylor & Francis Group, LLC; ISBN-13: 978-0-8493-4022-2.
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