The Tetracyclines Baran Lab D. W.

Lin
1
Cl
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
oxytetracycline
(terramycin)
1949
chlortetracycline
(aureomycin)
1948
The tetracycline family of antibiotics
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
tetracycline
(achromycin)
1953
Cl
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
HO
demethylchlortetracycline
(declomycin)
1957
OH O HO O
OH
O
H
N
NMe
2
OH
OH
H H
Me
HO
rolitetracycline
(reverin)
1958
OH O HO O
OH
O
H
N
NMe
2
OH
OH
H H
Me
HO
limecycline
(tetralysal)
1961
Cl
OH O HO O
OH
O
H
N
NMe
2
OH
H H
Me
HO
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
methacycline
(rondomycin)
1965
clomocycline
(megaclor)
1963
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
doxycycline
(vibramycin)
1967
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
minocycline
(minocin)
1972
NMe
2
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
t-butylglycylamidominocycline
(tigilcycline)
1993
(Phase III clinical trials in progress)
N HN
CO
2
H H
2
N
4
OH
OH
N
H
O
H
N
Me
Me
Me
The natural products
Semisynthetic derivatives on the market
Cl
OH O HO O
OH
O
NH
2
NMe
2
OH Me
HO
A B C D
1
2
3
4
5
6
7
8
9
10 11 12
Notation
OH
Me OH NMe
2
I. Chopra, M. Roberts. Microbiol. Mol. Biol. Rev. 2001, 65, 232.
4a 5a
12a
The Tetracyclines Baran Lab D. W. Lin
2
Cl
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
terramycin
aureomycin
Discovery and The Dawn of Semisynthetic Antibiotics
OH
Benjamin Duggar
University of Missouri
Bayer Pharmaceuticals
The first tetracycline antibiotic
discovered, aureomycin was
isolated in 1948 from a Missouri
soil sample by Lederle
Laboratories. The Lederle team
was led by Benjamin Duggar - a
consultant who was a 73-year-old
emeritus professor of botany who
had recently retired from the
University of Missouri! As Jie
Jack Li cracks, "Your greatest
discovery could happen in your
retirement."
About Lederle Labs:
Lederle Labs was founded in
1902 in an old farmhouse on the
Pearl River in New York.
Aureomycin was one of many
lifesaving products developed by
Lederle, including vaccines for
polio and smallpox. It is now a
part of Wyeth Pharmaceuticals.
R. B. Woodward
The Nobel Prize Committee
About the same time as the Lederle discovery of aureomycin, Pfizer was
scouring the globe for new antibiotics. Soil samples were collected from
jungles, deserts, mountaintops, and oceans. But ultimately terramycin was
isolated in 1949... from a soil sample collected on the grounds of a factory in
Terre Haute, Indiana, owned by Pfizer!
From the beginning, terramycin was a molecule enveloped in controversy. It
was the subject of the first mass-marketing campaign by a modern
pharmaceutical company. Pfizer advertised the drug heavily in medical
journals, eventually spending twice as much on marketing as it did to discover
and develop terramycin. Still, it turned Pfizer - then a small company - into a
pharmaceutical giant.
Pfizer and R.B. Woodward collaborated to determine the structure of
terramycin, succeeding for the most part in 1952 (JACS 1953, 75, 5455). The
stereochemistry at C
4a
was revised after X-ray crystallography and NMR
studies in the 1960's (JACS 1965, 87, 134; JACS 1963, 85, 851).
Ebay
Ad for aureomycin as additive in cattle feed
J. J. Li. "A History of Drugs and Their Discoverers." Pfizer Intranet Magazine. March-April 2004.
The Tetracyclines Baran Lab D. W. Lin
3
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
Cl
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
H
2
, Pd/C
MeOH/
dioxane
Big Pharma Behaving Badly: In 1955, Conover
discovered that hydrogenolysis of aureomycin gives a
deschloro product that is just as active as the original
product. This proved for the first time that chemically-
modified antibiotics could have biological activity. Within a
few years, a number of semisynthetic tetracyclines had
entered the market, and now most antibiotic discoveries are
of novel active derivatives of older compounds.
Conover's discovery, however, provoked further controversy
for tetracycline. Pfizer became embroiled in a patent
dispute with American Cyanamid, which owned the rights to
aureomycin (the starting material for Conover's procedure
to make tetracycline). Pfizer and American Cyanamid
eventually settled the dispute out of court when they
realized that neither company held truly exclusive rights to
the drug, and agreed to cooperate on selling the drug in
order to drive off competitors trying to enter the tetracycline
market. At one point, Pfizer employed a private decective
to tap the phones of Bristol-Meyers, a competitor seeking to
enter the tetracycline market! Bristol-Meyers agreed to
overlook this brazen act in exchange for a share of the
tetracycline market. Eventually five companies colluded in
order to maintain artificially high prices for tetracycline.
However, the Federal Trade Commission stepped in after
several years, finding Pfizer and company guilty of patent
fraud and anti-trust violations, and broke up the monopoly.
H
Conover, L.H. 1955. U.S. Patent No. 2,699,054.
Now, Back to Actual Science
Biosynthesis and Biological Activity: Tetracyclines are polyketide antibiotics,
biosynthesized in a fashion similar to that of fatty acids, erythromycin and a host of
other antibiotics. Tetracyclines are produced naturally by Streptomyces
aureofaciens (T. Nakano, et al. Biosci. Biotechnol. Biochem. 2004, 68, 1345.).
Tetracyclines bind to the bacterial ribosome, preventing the binding of aminoacyl-
tRNA to the ribosomal A site. This prevents bacterial protein translation (I. Chopra,
M. Roberts. Microbiol. Mol Biol. Rev. 2001, 65, 232.).
The Challenge to Synthetic Chemists: Muxfeldt and colleagues outline the basic
obstacles to achieving a total synthesis of any of the natural tetracyclines:
Stereochemical Complexity. There are up to five contiguous asymmetric centers
(terramycin) which must be established.
Chemical Sensitivity. For the 6-methyl-6-hydroxy tetracyclines, mild acid rapidly
catalyzes dehydration, ketalization and a retro-aldol to produce the lactone below.
Mildly basic conditions results in deprotonation of the C5 and C6 hydroxyls,
initiating a cascade of events which leads to decomposition of the molecule.
Finally, the C4 stereocenter is readily epimerized upon exposure to acetic acid or
aqueous buffers.
U.S. Federal Trade Commission, "Anticipating the 21st Century: Competition
Policy in the New High-Tech, Global Marketplace".
M. Mintz. "Golden Ox of Antitrust." The Nation 14 April 1969, Vol. 208, Issue 15.
pp. 467-468.
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
OH
acid
OH OH O O
OH
O
NH
2
NMe
2
OH
H
Me
O
OH O HO O
OH
O
NH
2
NMe
2
OH
H H
Me
HO
OH
acid
base
HO
COOH
Me
O
COOH
H. Muxfeldt, et al. J. Am. Chem. Soc. 1979, 101, 689.
About.com
Lloyd Conover
Pfizer
Legal issues aside, for this discovery Lloyd Conover is now in the American
Inventors' Hall of Fame, alongside Thomas Edison and the Wright brothers.
The Tetracyclines Baran Lab D. W. Lin
4
Woodward's First Total Synthesis of a Biologically-Active Tetracycline, 6-
Demethyl-6-deoxytetracycline.
L.H. Conover, et al. J. Am. Chem. Soc. 1962, 84, 3222. (Initial communication)
R.B. Woodward. Pure Appl. Chem. 1963, 6, 561. (A personal account)
J.J. Korst, et al. J. Am. Chem. Soc. 1968, 90, 439. (Full article)
CO
2
Me
OMe OMe
O O
OMe
O
OMe
NaH
DMF
O
OMe
O
OMe
NaH
DMF
Cl
(i)
(ii)
O
OMe
Triton-B
dioxane-MeOH
50-70
o
C
88%
OMe
O
CO
2
Me
CO
2
Me
CO
2
Me
55%
OMe
O
CO
2
Me
CO
2
Me
(i) AcOH/
H
2
SO
4
(ii) H
2
SO
4
MeOH/CHCl
3
44%
OMe
CO
2
Me
(i) H
2
, 200 psi
Pd/C
AcOH, 30
o
C
CO
2
Me
(ii) H
2
SO
4
MeOH/CHCl
3
93%
(i) NaOH
H
2
O
100
o
C
(ii) I
2
, AcOH;
then Cl
2
in AcOH
(iii) HF, neat
63% over 3 steps
CO
2
H
O OMe
Cl
Chlorination blocks the para
position, forcing condensation onto
the more hindered ortho position.
H
2
SO
4
MeOH/CHCl
3
reflux
66%
CO
2
Me
O OMe
Cl
CO
2
Me
O OMe
Cl
O O
MeO OMe
then NaH (4 eq.),
then MeOH (1 eq.),
rt --> 80
o
C
45%
(2 eq.)
CO
2
Me
O OMe
Cl
OH
OH
MeOH is essential to suppressing the kinetically
favorable intramolecular condensation and
permitting the intermolecular condensation with
the oxalate prior to formation of the desired
tricycle. In the absence of MeOH, Woodward
observed formation of the intramolecular product
first, followed by condensation onto the oxalate
to form the five-membered ring shown:
OH OMe
Cl
intermolecular condensation outcompetes intramolecular!
O
CO
2
Me
OH
CO
2
Me
O OMe
Cl
OH
OH
AcOH/HCl
H
2
O
90
o
C
73%
O OMe
Cl
OH
O
H CO
2
n
Bu
O
O OMe
Cl
OH
O
CO
2
n
Bu
O OMe
Cl
OH
OH
NMe
2
CO
2
n
Bu
H H
(i) NHMe
2
, -10
o
C;
(ii) NaBH
4
, -70
o
C
69%
Mg(OMe)
2
toluene
reflux
52%
The thermodynamically more favorable diastereomer is formed
exclusively in this step, with the carboxyamino substituent assuming
an equatorial position and thus establishing the cis relationship of
the bridgehead hydrogens. Ketone reduction is also stereoselective.
The Tetracyclines Baran Lab D. W. Lin
5
O OMe
Cl
OH
OH
NMe
2
CO
2
n
Bu
H H
TsOH
toluene
reflux
90% O OMe
Cl
OH
H H
O
NMe
2
O
Zn dust
formic acid
1 min
81%
H
2
, Pd/C
Et
3
N
91%
O OMe OH
NMe
2
CO
2
H
H H
(i)
O
O
Cl
(ii)
EtO
O O
NH
t
Bu
Mg(OEt)
O OMe OH
NMe
2
CO
2
H
H H
Cl
O OMe OH
NMe
2
H H
OH
EtO
2
C
O
NH
t
Bu
NaH
DMF/MeOH
120
o
C
15 min
OH OMe OH
NMe
2
H H
O
OH
CONH
t
Bu
A No acylation of the enols by the
chloroformate was observed.
O OH OH
NMe
2
H H
OH
O
CONH
2
OH
O
2
CeCl
3
DMF-MeOH
glycine-NaOH buffer,
pH = 10
15 min
Mixture of epimers at C
4
CaCl
2
BuOH-H
2
O, pH = 8.5
ethanolamine buffer
reflux, 10 min
6% over 2 steps,
10% recovered SM
O OH OH
NMe
2
H H
OH
O
CONH
2
OH
Thermodynamic equilibration
to the desired epimer.
6-desmethyl-6-deoxytetracycline
This was the first total synthesis of a tetracycline with all the requisite
functionality for full antibiotic activity. Note, however, that this is not the total
synthesis of a tetracycline natural product. Substituents at the C
6
position are
missing.
O OMe O
NMe
2
H H
O
O
N
t
Bu
O
EtO
Observe the classic Woodward master
stroke. Despite the presence of four
enolates, we observe only one of two
plausible intramolecular condensation
events. The other event is impossible since
the enolate double bond cannot rotate to
bring the amide into position for cyclization.
48% HBr
20 min
OH OH OH
NMe
2
H H
O
OH
CONH
2
15% bsm from A;
30% of A recovered
OH OMe OH
NMe
2
H H
O
OH
CONH
t
Bu
6
This was a difficult step to optimize - Woodward himself noted dryly that "the case
at hand was by no means the smoothest we had encountered." Competitive
hydroxylation at C
11a
was also observed, as well as rapid decomposition of the
product under prolonged reaction conditions, forcing Woodward and colleagues
to halt the reaction prematurely.
The Tetracyclines Baran Lab D. W. Lin
6
Shemyakin: The First Total Synthesis of a Tetracycline Natural Product
A.I. Gurevich, et al. Tet. Lett. 1967, 8, 132.
M.N. Kolosov, S.A. Popravko, M.M. Shemyakin. Lieb. Ann. 1963, 668, 86.
B.-M.G. Gaveby, J.C. Huffmann, P. Magus. J. Org. Chem. 1982, 47, 3779.
Note that the Lieb. Ann. reference cites a number of obscure Russian journals.
The JOC reference, however, illustrates Shemyakin's approach to the tricyclic
precursor produced below.
Me
H
HO
O OH OBn
O
2
N
O
OEt
Et
3
NH
+
THF
Me
H
HO
O OH OBn
CO
2
Et
NO
2
HCl
EtOH
OH O OBn
CO
2
Et
NO
2
Me
Zn
dust
AcOH
OH O OBn
CO
2
Et
NH
2
Me
OMe O OBn
CO
2
Et
NPhth Me
N
O
O
O
OEt
(i)
(ii) MeI, Ag
2
O
H
H H
H
(i) 0.1 N KOH, THF-H
2
O
N
O
O
O
OEt
(ii)
OMe O OBn
CO
2
H
NPhth Me
H
PCl
5
in DMF, then
EtO
O O
NH
2
Mg(OEt)
OMe O OBn
NPhth Me
H
OH
EtO
2
C
CONH
2
Notice Shemyakin adopting the Woodward approach to ring A.
O
O OH
OAc
+
BF
3
.
OEt
2
O
O OH OAc
H
H
86%
LiAl(O
t
Bu)
3
H
64%
O
OH OH OAc
H
H
BnBr
K
2
CO
3
54%
O
OH BnO OAc
H
H
MeMgBr
6 eq
74%
OH
OH BnO OAc
H
H Me
KOH/MeOH
85%
OH
OH BnO OH
H
H Me
Jones reagent
60%
Me
H
HO
O OH OBn
The Tetracyclines Baran Lab D. W. Lin
7
S
O
Na
+
DMSO
OMe O OBn
Me
H
HN
HO
2
C
OH
OH
CONH
2
(i) HBr-AcOH
(ii) MeI in THF
OH O OH
Me
H
NMe
2
OH
OH
CONH
2
This intercepts a degradation product which had
previously been elaborated into tetracycline.
O
2
over Pt
Et
3
N
THF
rt, 8 hr
OH O OH
Me
H
NMe
2
OH
O
CONH
2
OH
A.I. Gurevich, M.G. Karapetyan,
M.N. Kolosov. Khim. Prirodn.
Soedin., Akad. Nauk Uz.SSR
1966, 141.
(i) O
2
, h
3,4-benzopyrene (cat.)
benzene
(ii) H
2
, Pd/C
O OH OH
H
NMe
2
OH
O
CONH
2
OH
M. Schach von Wittenau. J. Org. Chem. 1964, 29, 2746.
H
Me
HO
tetracycline
Mechanism? Answer on Slide 16.
OMe O OBn
NPhth Me
H
OH
EtO
2
C
CONH
2
The Tetracyclines Baran Lab D. W. Lin
8
Muxfeldt's Total Synthesis of 6-Desmethyl-6-deoxytetracycline
H. Muxfeldt, W. Rogalski. J.Am. Chem. Soc. 1965, 87, 933. (Communication)
H. Muxfeldt, E. Jacobs, K. Uhlig. Chem. Ber. 1962, 95, 2901. (Prep of precursors)
Cl
OMe
Br
NaOMe
MeOH
Cl
OMe
CO
2
Me
CO
2
Me
CO
2
Me
(i) NaOH
(ii) pyrolysis,
160
o
C
85%
over three steps
Cl
OMe
CO
2
H
CO
2
H
H
3
PO
4
O
CO
2
H
Cl
OMe
80
o
C
quant.
CO
2
Me
Cl
MeO
O O
Cl
MeO
O O
OH
Cl
MeO
O O
CN
Cl
MeO
O O
CHO
(i) H
2
SO
4
MeOH
95%
(ii) ethylene glycol
TsOH, benzene
91%
LiAlH
4
benzene-Et
2
O
0
o
C
94%
(i) MsCl
pyridine
97%
(ii) NaCN
NaI (cat.)
DMF-H
2
O
92%
Li(EtO)
3
AlH
benzene-Et
2
O
0
o
C
64%
HCl
THF
C
6
H
5
O
N
H
CO
2
H
Pb(OAc)
4
(cat.)
Ac
2
O
Cl
MeO
N
O
Ph
O
O
Cl
MeO
N
O
Ph
O
O O
MeO
2
C
O O
NH
t
Bu
Cl
MeO
N
O
Ph
O
NaH (2 eq.)
THF-Et
2
O
35
o
C, 24 hr
MeO
2
C
O
CONH
t
Bu
O
Cl
MeO
N
O
Ph
O
MeO
2
C
O
CONH
t
Bu
O
Cl
MeO
N
O
MeO
2
C
O
CONH
t
Bu
O
Ph
O
Cl
MeO
N
O
MeO
2
C
O
CONH
t
Bu
O
O
Ph
Cl
MeO
HN
O
MeO
2
C
O
CONH
t
Bu
O
Now the stage is set for the second cyclization in this magnificent transformation.
Only one equivalent of NaH used so far!
O
Ph
taut.
taut.
MeO
2
C CO
2
Me
CO
2
Me
The Tetracyclines Baran Lab D. W. Lin
9
Cl
MeO
HN
O
MeO
2
C
O
CONH
t
Bu
O
O
Ph
NaH
Cl
MeO
NHBz
O O
CONH
t
Bu
O
MeO
O
Cl
MeO
NHBz
O O
CONH
t
Bu
O
O
Cl
MeO
NHBz
O O
CONH
t
Bu
OH
OH
82% from the starting aldehyde
isolated as mixture of epimers
at C
4
Muxfeldt thus effects the construction of the A and B rings in a single step! The only
problem, unfortunately, is the failure to control C
4
stereochemistry.
(i) Me
3
OBF
4
(ii) HBr/AcOH,
100
o
C
Cl
OH
NH
2
O O
CONH
2
OH
OH
H
2
, Pd/C, H
2
CO
Et
3
N, MeOH
OH
NMe
2
O O
CONH
2
OH
OH
OH
NMe
2
O O
CONH
2
OH
OH
OH
6-Demethyl-6-deoxytetracycline
Here they intercept an intermediate from the Woodward synthesis. They also report
hydroxylation with O
2
over platinum (Angew. Chem. Intl. Ed. Eng. 1962, 1, 157).
(i) deprotects the benzoyl amide; (ii) deprotects the remaining functional groups.
Muxfeldt's Last Hurrah: Total Synthesis of Terramycin
H. Muxfeldt, et al. J. Am. Chem. Soc. 1968, 90, 6534.
H. Muxfeldt, et al. J. Am. Chem. Soc. 1979, 101, 689.
Terramycin is a much more difficult target than the prototypical tetracyclines
discussed previously - Woodward and Muxfeldt avoided many of the problems
outlined earlier with by targeting a structure without the troublesome C
5
and C
6

substituents, while Shemyakin targeted a tetracycline which did not have the
C
6
hydroxyl. Here Muxfeldt and colleagues (including a young Edwin Vedejs!)
tackle those problems head-on! Sadly, this is reported in a posthumous
communication from the Muxfeldt laboratories.
OH
O
O
(i) Ac
2
O, H
2
SO
4
0
o
C
83%
(ii) 1-acetoxy-
butadiene
benzene, reflux
60%
AcO
O
O
H
H
OAc (i) MeMgI,
-65
o
C
82%
(ii) NaOH
84%
HO
OH
O
H
H
OH
Me
AcO
O
O
H
H
O
Me
(i) acetone,
CuSO
4
84%
(ii) Ac
2
O,
NaOAc
95%
Me Me
KClO
3
OsO
4
(cat.)
50
o
C
89%
AcO
O
O
H
H
O
Me
Me Me
OH
OH
Mixture of cis-diols
Pb(OAc)
4
40
o
C
AcO
O
O
H
H
O
Me
Me Me
O
O
The Tetracyclines Baran Lab D. W. Lin
10
AcO
O
O
H
H
O
Me
Me Me
O
O
AcO
O
O
H
H
O
Me
Me Me
CHO
DBU-AcOH,
piperidine
(cat.)
xylenes
reflux
52% over
two steps
(i) O
3
-50
o
C
(ii) H
2
O
68%
AcO
O
O
CHO
H
H
O
Me
Me Me
CHO
O HO
O
O
CHO
H
O
Me
Me Me
Mixture of C
5
epimers
2 : 1
CH
2
Cl
2
:
0.5 N NaCO
3
in H
2
O
85%
H
N
(i)
(ii) NaH, then
MOMCl
90%
MOMO
O
O
H
O
Me
Me Me
N
silica gel,
deactivated
91%
70-80%
MOMO
O
O
CHO
H
O
Me
Me Me
N
S
O
Ph
Pb
2
(OH)(OAc)
3
77%
MOMO
O
O
H
O
Me
Me Me
The thermodynamically more favorable epimer is obtained exclusively.
S
N
O
Ph
B
Preparation of C:
MeO
O O O
OMe
NH
3
MeOH
33%
MeO
O NH
2
O
NH
2
conc. HCl
MeO
O O O
NH
2
CHCl
3
60%
C
Coupling of B and C:
MeO
O O O
NH
2
MOMO
O
O
H
O
Me
Me Me
S
N
O
Ph
BuLi (1.0 eq), -78
o
C,
then
THF, reflux
2 h
MOMO
O
O
H
O
Me
Me Me
OH
H
O
CONH
2
OH
HN
S
Ph
H
MOMO
O
O
H
O
Me
Me Me
MeO
2
C
O
CONH
2
OH
HN
S
Ph
H
27%
+
Mixture of diastereomers at C
4
, C
4a
Once again, Muxfeldt employs his beautiful method for forming the A and B
rings in a single step. And once again, there is little stereocontrol - all four
possible epimers at C
4
and C
4a
are formed in solution. Fortunately, the desired
diastereomer readily crystallizes. The reason for employing the thiazolanone
rather than the oxazolanone employed before will become clear shortly...
The Tetracyclines Baran Lab D. W. Lin
11
MOMO
O
O
H
O
Me
Me Me
OH
H
O
CONH
2
OH
HN
S
Ph
H
HO
O
O
H
O
Me
Me Me
OH
H
O
CONH
2
OH
HN
S
Ph
H
9 : 1
AcOH :
H
2
O
reflux,
6 min
90%
No epimerization at C
4
observed!
HO
OH
O
H
OH
Me
OH O
CONH
2
OH
HN
S
Ph
H
OH (i) P(OEt)
3
, NaH, O
2
DMF-THF-H
2
O
15 min
(ii) 0.01 N HCl
in MeOH
rt, 1.5 h
(i) hydroxylates the molecule; (ii) cleaves the acetonide. Unfortunately,
hydroxylation occurs principally at C
11a
. In a fortunate accident, however, it turned
out that the acetonide could not be cleaved unless the C
12a
hydroxyl was present.
Thus separation of the desired deprotected product from the undesired major
product was quite facile by polyamide chromatography.
(i) MeI in THF
(ii) 0.17 N HCl
in THF-H
2
O, 1.5 h
Cl
Me
2
SO
4
(i-Pr)
2
NEt
23% from
thioamide
HO
OH
O
H
OH
Me
OH O
CONH
2
OH
NMe
2
H
OH
terramycin
While the oxazolone substrate could also be carried to this step, the resulting
benzoyl amide could not be deprotected at this stage, nor could any other
amide devised, without decomposition. By contrast, deprotection conditions
for the thioamide proved to be sufficiently gentle.
HO
O
O
H
O
Me
Me Me
OH
OH
O
CONH
2
OH
HN
S
Ph
H
12%
desired C
12a

hydroxylated
product
47%
C
11a

hydroxylated
byproduct
14% recovered SM
HO
OH
O
H
OH
Me
OH O
CONH
2
OH
HN
S
Ph
H
OH
HO
OH
O
H
OH
Me
OH O
CONH
2
OH
NH
3
H
OH
+
+
This concludes an elegant synthesis which assembles the A and B rings in a
single step. Unfortunately, Muxfeldt and colleagues never satisfactorily address
the issues of controlling the C
4
and C
4a
stereocenters, nor do they improve
upon Woodward's solution to the C
12a
hydroxylation problem.
The Tetracyclines Baran Lab D. W. Lin
12
O OH
O
Me
O O
OAllyl
CHO
piperidine (11 eq)
AcOH (40 eq)
mol. sieves
benzene
0
o
C --> rt
2.5 h
97%
O OH
Me
O
O
O
OAllyl
45% overall yield from start
of the synthetic sequence!
O
N
MeO
2
C
OBn
Me
2
N
, then
NaHMDS, then
the above tricycle
O OH
Me
O
O
O
OAllyl
N
O
NMe
2
H
OBn
MeO
2
C
Here Stork exploits the stereochemistry of the tricycle to
direct conjugate addition to the more accessible face.
Observe that the C
5a
and C
4a
stereocenters are now set.
Pd(PPh
3
)
4
95% from
the tricycle
O OH
Me
O
O
N
O
NMe
2
H
OBn
MeO
2
C
5a
4a
5a
4a
Bu
3
SnOCH
3
60
o
C
97%
O OH
Me
OH
H H
5a 4a
MeO
2
C
N
O
OBn
MeO
2
C
NMe
2
TMSCN
KCN
18-crown-6
OTMS OTMS
H H
MeO
2
C
N
O
OBn
MeO
2
C
NMe
2
Me
O
This protects the C
6
and
C
10
hydroxyls, and sets
the stage for the
remaining cyclization
steps.
Stork: Controlling the C
4
, C
4a
Stereocenters
G. Stork, et al. J. Am. Chem. Soc. 1996, 118, 5304.
Here Gilbert Stork and colleagues take a completely different approach in order
to achieve stereocontrol at the C
4
and C
4a
centers.
O
O
O
O
(i) MeMgBr
78%
100%
(ii)
100%
O
OH OH
OH
Br
OEt
Br
N,N-dimethyl-
aniline,
98%
OH
Me
O OH
O
Me
OEt
Br
n-Bu
3
SnH,
AIBN

90%
O OH
Me
O
OEt
HS SH
BF
3
.
OEt
2
0
o
C, 15 min
88%
O OH
OH
Me
S
S
HO
O O
O
92%
O OH
O
Me
S
S
O O
OAllyl
Stork postulates a ketene intermediate
formed from the mixed anhydride.
TFA anhydride, then
, then
the dithiane
(i) PhI(OTFA)
2
,
MeOH
92%
(ii) 5% aq. HCl
quant.
O OH
O
Me
O O
OAllyl
CHO
Transketalization, followed
by hydrolysis to aldehyde.
This sets the C
6
stereocenter. Now watch Stork use this
stereocenter to bootstrap his way through the molecule...
Mild reagent for
lactone cleavage
The Tetracyclines Baran Lab D. W. Lin
13
OTMS OTMS
H H
MeO
2
C
N
O
BnO
MeO
2
C
NMe
2
Me
O
KH
(25 eq)
-78 --> 0
o
C,
3 h, then
0 --> 50
o
C,
30 min
OTMS OTMS
H H
MeO
2
C
N
O
BnO
NMe
2
Me
O
O
O O
H H
N
O
BnO
NMe
2
Me
O
O
O OH
H H
N
O
BnO
NMe
2
Me
OH
OH
OH
O
MeO
59%
The protecting group scheme permits formation of the A ring first, followed by in
situ deprotection and cyclization of the B ring to complete the basic tetracycline
framework. Previous studies had indicated that failure to protect the C
11
ketone
resulted in formation of a BCD tricycle for which conditions to complete A ring
cyclization could not be found.
H
2
Pd black
94%
O OH
H H
NMe
2
Me
OH
OH
OH
CONH
2
OH
12a-deoxytetracycline
A Note on C
12a
Hydroxylation: This intercepts an intermediate which has
been hydroxylated at the C
12a
position according to literature reports,
completing in principle the formal synthesis of tetracycline. However, Stork
and colleagues were unable to successfully apply any of the C
12a

hydroxylation methods previously reported. The presence of the C
4

dimethylamino substituent seems to interfere with the hydroxylation. Clearly
a satisfactory solution to the C
12a
hydroxylation problem is still needed...
The Tetracyclines Baran Lab D. W. Lin
14
Tatsuta: Asymmetric Total Synthesis of Natural (-)-Tetracycline
K. Tatsuta, et al. Chem. Lett. 2000, 647.
Here Tatsuta and colleagues not only produce an asymmetric total synthesis, but
they also take a very different approach to the synthetic problem, constructing
the A and B rings first and exploiting the carbohydrate chiral pool for starting
materials. And as a bonus, they solve the C
12a
hydroxylation problem as well!
O
HO
OMe
CbzHN
OTBS
BnO
O
OMe
CbzHN
OTBS
BnO
H
2
C
(i) DMSO
DCC, Py-TFA
97%
(ii) Ph
3
PCH
3
Br
BuLi/THF,
-78
o
C --> rt
91%
(i) HCl-MeOH
93%
NO
2
SeCN
(ii)
PBu
3
(cat.)
90%
O
OMe
CbzHN
Se
BnO
H
2
C
NO
2
BH
3
.
THF,
0 --> 45
o
C;
then H
2
O
2
,
NaOH/THF
69%
O
OMe
CbzHN
CH
2
BnO
HO
(i) BnBr
BaO/Ba(OH)
2
84%
(ii) HgCl
2
THF-H
2
O
67%
OH
CbzHN
O
BnO
BnO
OH
CHO
CbzHN
CH
2
BnO
BnO
(i) MsCl, Et
3
N
0
o
C, 15 min
82%
(ii) DBU, -30
o
C
quant.
In addition to eliminating to the enone, (ii) also
epimerizes to the thermodynamic diastereomer.
O
NHCbz
BnO
OBn
OTMS
OH
t
Bu
t
Bu
Me
170
o
C
72%
OH O
H
NHCbz
OBn
OBn
CrO
3
, H
2
SO
4
0
o
C, 10 min
85%
O OH
H
NHCbz
OBn
OBn
H
O
Me
O
LDA, -40
o
C,
15 min
80%
O OH
H
NHCbz
OBn
OBn OH
OMe
H
OH
Me
OMe
-30
o
C
10 min
90%
SOCl
2
Et
3
N
O OH
H
NHCbz
OBn
OBn OH OMe
Me
(i) BBr
3
-78
o
C
15 min
(ii) H
2
, Pd/C
Boc
2
O, Et
3
N
92% over
two steps
O OH
H
NHBoc
OH
OH OH OMe
Me
O OMe
H
NHBoc
OH
OH OH OMe
Me
Attempts to directly oxidize this 1,3-diol to the 1,3-dicarbonyl failed, requiring
the following detour of sequential alcohol oxidations.
TMSCHN
2
i-Pr
2
NEt
72%
The Tetracyclines Baran Lab D. W. Lin
15
O OMe
H
NHBoc
OH
OH OH OMe
Me
Br
2
(Bu
3
Sn)
2
O
mol. sieves
-78
o
C, 15 min
85%
O
H
NHBoc
OH
OH OMe
Me
O
Br
OMe
Dess-Martin
periodinane
Zn, AcOH
2 min
O
H
NHBoc
O
OH OMe
Me
OMe OH
H
Dess-Martin
periodindane
15 min
91%
O
H
NHBoc
O
OH OMe
Me
O
Br
OMe
15 min
62% over
two steps
O
H
NHBoc
O
OH OMe
Me
OMe
OH
O O
NTs
B
TsN
Cl
Ph Ph
Et
3
N
-78
o
C, 30 min
60%
O
H
NHBoc
OH
OH OMe
Me
O
O
OH
(i) H
2
NOH
.
HCl
Et
3
N, 30 min
O
N N
N
N
(ii)
80%over 2 steps
Mechanism?
Here Tatsuta et al. employ DMDO to achieve the desired
hydroxylation. They also achieve enantioselectivity by
exploiting the chiral boron catalyst which Corey developed
for enantioselective aldols and Diels-Alder reactions. Note
the fantastic yield!
N
OH
H
N
O
H
O
N
N
N
O
H
NH
2
OH
OH OMe
Me
O
CN
OH
O
H
NMe
2
OH
OH OMe
Me
O
CONH
2
OH
(i) H
3
PO
4
100
o
C,45 min
68%
(ii) H
2
CO,
HCOOH
80
o
C, 1 h
80%
O
H
NMe
2
OH
OH OH
Me
O
CONH
2
OH
BBr
3
0 --> rt
88%
O
2
, h
meso-tetraphenyl-
porphyrin (cat.)
10 min
75%
O
H
NMe
2
OH
O OH O
CONH
2
OH
Me
OOH
Here Tatstuta et al apply a protocol developed by Wassermann, Lu and
Scott for hydroxylating anhydrotetracyclines. Provide a mechanism for this
reaction, and rationalize the stereospecific nature of this reaction.
H. Wassermann, T.-J. Lu, A.I. Scott. J. Am. Chem. Soc. 1986, 108, 4237.
The Tetracyclines Baran Lab D. W. Lin
16
O
H
NMe
2
OH
O OH O
CONH
2
OH
Me
OOH
H
2
/Pt
62%
OH
H
NMe
2
OH
O OH O
CONH
2
OH
Me
OH
H
(-)-tetracycline
O
O
H
H
H
H
NMe
2
O
H
O
N
H
O
H
OH
H
O O
O
O
H
H
NMe
2
O
H
O
N
H
O
H
OH
H
H
H
O
H
Wassermann, Lu and Scott invoke a formal ene reaction. The orbital
alignment requirements dictate that only the axial hydrogen can participate
in the reaction, inducing hydroperoxidation on the upper face of the
molecule and thus ensuring the proper stereochemistry at C
6
.
O
H
There are many elegant features to this synthesis. Tatsuta and colleagues
mimic Stork's Diels-Alder approach to establishing stereochemistry, but employ
it to define the troublesome C
4a
stereocenter immediately. They construct the
central tetracycline scaffolding in just three steps from simple precursors. And
they solve the C
12a
hydroxylation problem with a very mild oxidant in the
presence of a chiral catalyst, and introduce the C
6
hydroxyl stereospecifically
at a very late stage of the synthesis.
Myers' Rapid Asymmetric Access to Analogs of Tetracycline
M.G. Charest, C.D. Lerner, J.D. Brubaker, D.R. Siegel, A.G. Myers. Science 2005,
308, 395.
In an extraordinary report, Myers and colleagues present a highly efficient and
enantioselective method for accessing the tetracyclines.
CO
2
H
A. eutrophus B9
75%, >95% ee
CO
2
H
OH
OH
mCPBA
EtOAc
83%
CO
2
H
OH
OH
O
This bacterial-catalyzed reaction
can be run on a 90 g scale!
1. TMSCHN
2
2. TBSOTf, Et
3
N
OTBS
TBSO
CO
2
Me
O
N
O
NMe
2
OBn
Li
THF, -78
o
C
OTBS
TBSO
O
O
O
N
NMe
2
OBn
70%
73%
1. LiOTf,
toluene, 60
o
C
2. TFA, CH
2
Cl
2
60%
N
O
OH
TBSO
TBSO
N
O
Me
Me
Here Myers closes the ring and sets the C
4
amine stereochemistry. Myers
compares this key ring-closing step to a Sommelet-Hauser rearrangement, where
the amine initially undergoes an intramolecular S
N
-prime epoxide ring opening,
followed by ylide formation and finally a [2,3] sigmatropic rearrangement. TFA
selectively deprotects the allylic alcohol. Notice the remarkable yield so far!
N
O
OH
O
BnO
OBn
TBSO
HO
H
NMe
2
12a
A B
21% over 7 steps
Notice how Myers begins with installation of the
troublesome C
12a
hydroxyl group, and then
proceeds to build the molecule around it!
The Tetracyclines Baran Lab D. W. Lin
17
N
O
OH
O
OBn
TBSO
HO
H
NMe
2
PPh
3
, DEAD;
then
O
2
S
N
H
NH
2
NO
2
N
O
OH
O
OBn
TBSO
H
NMe
2
1. HCl, MeOH
2. IBX, DMSO
3. TBSOTf, 2,6-lutidine
74%
66%
N
O
OTBS
O
OBn
O
H
NMe
2
A
B
N
O
OH
O
OBn
TBSO
HO
H
NMe
2
A
Now Myers takes his key intermediate A and converts it into two fragments: B,
which will go on to form C
6
-deoxy analogs of tetracycline, and C, which will go
on to form analogs with the normal C
6
-oxygenation.
1. CBr
4
, PPh
3
2. PhSH, Et
3
N
87%
N
O
OH
O
OBn
TBSO
PhS
H
NMe
2
O
N
S
O
2
Me Me
Cl
Cl
1.
2. P(OMe)
3
, MeOH
70
o
C
N
O
OH
O
OBn
TBSO
H
NMe
2
OH
76%
1. BnO
2
CCl, DMAP
2. TBAF, HOAc
3. IBX, DMSO
4. TBSOTf, Et
3
N
85%
N
O
OTBS
O
OBn
O
H
NMe
2
BnO
2
CO
C
With these fragments in hand, Myers now can install the C and D rings, and he
proceeds to do so in a fashion that allows for analogs of tetracycline with deep-
seated structural modifications to the D ring.
Me
CO
2
Ph
BocO
LDA, TMEDA,
THF, -78
o
C;
then C,
-78
o
C -> 0
o
C
79%
N
O
OTBS
O
OBn
OH
H
NMe
2
O
O
H
Me
O
OBn
OH
1. HF, MeCN
2. H
2
, Pd, THF/MeOH
90%
O OH
H
NMe
2
OH
O
H
Me
OH
OH
O
NH
2
OH
(-)-doxycycline
18 steps, 8.3%
The Tetracyclines Baran Lab D. W. Lin
18
With this strategy, Myers and colleagues are able to synthesize a number of
remarkable analogs of tetracycline:
OBoc
CO
2
Ph
Me
O OH
H
NMe
2
O
H
Me
OH
OH
O
NH
2
OH
B
N
OBn
CO
2
Ph
Me Me
B
O OH
H
NMe
2
HN
O
H
OH
O
NH
2
OH
O
(-)-6-deoxytetracycline
N
CO
2
Ph
Me
B
O OH
H
NMe
2
N
O
H
OH
O
NH
2
OH
CH
2
Br
CO
2
Ph
B
O OH
H
NMe
2
O
H
OH
O
NH
2
OH
OMe
CO
2
Ph
CH
2
Br
B
O OH
H
NMe
2
O
H
OH
O
NH
2
OH
OMe
The Tetracyclines Baran Lab D. W. Lin
19
Addendum: Tetracycline Tidbits
D.H.R. Barton spent over a decade tinkering with tetracycline, but never completed
a total synthesis of the molecule. Over the course of this work, however, he
discovered some interesting chemistry (naturally).
Photocyclizations of acetals onto enones:
In his book Reason and Imagination, Barton concludes his chapter on the
tetracyclines with the following perspective on the role of academic research in
synthetic chemistry today:
"Just as the studies on the bitter principles [a class of natural products]
convinced me that X-ray crystallography was a superior procedure for structure
determination, the major effort on tetracycline synthesis convinced me that this
sort of work should be left to Industrial friends who have the money and the
resources to finish any multi-step synthesis, if it is economically justified. So it
is the originality in the reactions and the reagents and any new principles
that finally justify academic effort in synthesis. We are far away from the
Woodwardian dogma of completely planned synthesis."
O
O
H
O
Ph
O
h
ArCO
2
H
O
Ph
O
O O
H H
O
O
H
O
Ph
O
O
O
O
Ph
OH
Barton and colleagues (including a young Steven Ley!) also discovered the
utility of phenylseleninic anhydride for the deprotection of dithianes. This led to
their applying this reagent in a variety of transformations:
D.H.R. Barton, et al. J. Chem. Soc. Perkin Trans. I 1976, 503.
AcO
Me
H
O
Me
R
R'
PhSe
O
O
SePh
O
AcO
Me
H
O
Me
R
R'
D.H.R. Barton, D.J. Lester, S.V. Ley. J. Chem. Soc. Perkin Trans. I 1980, 2209.