Baran Lab The Tetracyclines D. W.

Lin

Cl HO Me NMe2 OH NMe2
The tetracycline family of antibiotics H H H H
OH OH

The natural products H

N NH2
Cl HO Me NMe2 HO Me OH NMe2 OH OH
H H H H OH O HO O O OH OH O HO O O
OH OH
clomocycline methacycline
NH2 NH2 (megaclor) (rondomycin)
1963 1965
OH OH
OH O HO O O OH O HO O O Me OH NMe2 NMe2 NMe2
chlortetracycline oxytetracycline H H H H
OH OH
(aureomycin) (terramycin)
1948 1949
NH2 NH2
HO Me NMe2 Cl HO NMe2 OH OH
H H H H OH O HO O O OH O HO O O
OH OH
doxycycline minocycline
NH2 NH2 (vibramycin) (minocin)
1967 1972
OH OH
OH O HO O O OH O HO O O NMe2 NMe2
H H
tetracycline demethylchlortetracycline OH
(achromycin) (declomycin) O
H
1953 1957 Me N NH2
N
H OH
Me Me OH O HO O O
Semisynthetic derivatives on the market t-butylglycylamidominocycline
(tigilcycline)
1993
HO Me OH NMe2 HO Me OH NMe2
(Phase III clinical trials in progress)
H H H H
OH OH
H H Cl HO Me OH NMe2
N N
7 5
6 5a 4a 4 3 OH
OH OH 8
OH O HO O O N OH O HO O O HN D C B A 2
9 NH2
4
rolitetracycline limecycline 10 11 12 12a 1
(reverin) (tetralysal) H2N CO2H OH O HO O O
1958 1961 Notation

I. Chopra, M. Roberts. Microbiol. Mol. Biol. Rev. 2001, 65, 232. 1

 Baran Lab The Tetracyclines D. W. Lin

Discovery and The Dawn of Semisynthetic Antibiotics HO Me OH NMe2

H H
OH

NH2
OH
Cl HO Me NMe2 OH O HO O O
H H terramycin
OH

NH2
OH
OH O HO O O
aureomycin
Bayer Pharmaceuticals

Benjamin Duggar
University of Missouri

The first tetracycline antibiotic

discovered, aureomycin was
isolated in 1948 from a Missouri The Nobel Prize Committee
soil sample by Lederle R. B. Woodward
Laboratories. The Lederle team
was led by Benjamin Duggar - a About the same time as the Lederle discovery of aureomycin, Pfizer was
consultant who was a 73-year-old scouring the globe for new antibiotics. Soil samples were collected from
emeritus professor of botany who jungles, deserts, mountaintops, and oceans. But ultimately terramycin was
had recently retired from the isolated in 1949... from a soil sample collected on the grounds of a factory in
University of Missouri! As Jie Terre Haute, Indiana, owned by Pfizer!
Jack Li cracks, "Your greatest
discovery could happen in your
retirement." From the beginning, terramycin was a molecule enveloped in controversy. It
was the subject of the first mass-marketing campaign by a modern
About Lederle Labs: pharmaceutical company. Pfizer advertised the drug heavily in medical
Lederle Labs was founded in journals, eventually spending twice as much on marketing as it did to discover
1902 in an old farmhouse on the and develop terramycin. Still, it turned Pfizer - then a small company - into a
Pearl River in New York. pharmaceutical giant.
Aureomycin was one of many
lifesaving products developed by Pfizer and R.B. Woodward collaborated to determine the structure of
Lederle, including vaccines for terramycin, succeeding for the most part in 1952 (JACS 1953, 75, 5455). The
polio and smallpox. It is now a
stereochemistry at C4a was revised after X-ray crystallography and NMR
part of Wyeth Pharmaceuticals.
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. 2
 Baran Lab The Tetracyclines D. W. Lin

Big Pharma Behaving Badly: In 1955, Conover Now, Back to Actual Science
discovered that hydrogenolysis of aureomycin gives a
deschloro product that is just as active as the original Biosynthesis and Biological Activity: Tetracyclines are polyketide antibiotics,
product. This proved for the first time that chemically- biosynthesized in a fashion similar to that of fatty acids, erythromycin and a host of
modified antibiotics could have biological activity. Within a other antibiotics. Tetracyclines are produced naturally by Streptomyces
few years, a number of semisynthetic tetracyclines had aureofaciens (T. Nakano, et al. Biosci. Biotechnol. Biochem. 2004, 68, 1345.).
entered the market, and now most antibiotic discoveries are
of novel active derivatives of older compounds. Tetracyclines bind to the bacterial ribosome, preventing the binding of aminoacyl-
tRNA to the ribosomal A site. This prevents bacterial protein translation (I. Chopra,
Conover's discovery, however, provoked further controversy M. Roberts. Microbiol. Mol Biol. Rev. 2001, 65, 232.).
for tetracycline. Pfizer became embroiled in a patent
dispute with American Cyanamid, which owned the rights to The Challenge to Synthetic Chemists: Muxfeldt and colleagues outline the basic
aureomycin (the starting material for Conover's procedure obstacles to achieving a total synthesis of any of the natural tetracyclines:
to make tetracycline). Pfizer and American Cyanamid
eventually settled the dispute out of court when they Stereochemical Complexity. There are up to five contiguous asymmetric centers
realized that neither company held truly exclusive rights to (terramycin) which must be established.
the drug, and agreed to cooperate on selling the drug in
order to drive off competitors trying to enter the tetracycline Chemical Sensitivity. For the 6-methyl-6-hydroxy tetracyclines, mild acid rapidly
market. At one point, Pfizer employed a private decective catalyzes dehydration, ketalization and a retro-aldol to produce the lactone below.
to tap the phones of Bristol-Meyers, a competitor seeking to Mildly basic conditions results in deprotonation of the C5 and C6 hydroxyls,
enter the tetracycline market! Bristol-Meyers agreed to About.com initiating a cascade of events which leads to decomposition of the molecule.
overlook this brazen act in exchange for a share of the Finally, the C4 stereocenter is readily epimerized upon exposure to acetic acid or
Lloyd Conover aqueous buffers.
tetracycline market. Eventually five companies colluded in Pfizer
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. HO Me OH NMe2 Me NMe2
H H H
OH OH
Legal issues aside, for this discovery Lloyd Conover is now in the American acid
Inventors' Hall of Fame, alongside Thomas Edison and the Wright brothers. O
NH2 NH2
U.S. Federal Trade Commission, "Anticipating the 21st Century: Competition OH OH
Policy in the New High-Tech, Global Marketplace". OH O HO O O OH OH O O O

acid base
M. Mintz. "Golden Ox of Antitrust." The Nation 14 April 1969, Vol. 208, Issue 15.
pp. 467-468. O
Cl HO Me NMe2 H HO Me NMe2 HO Me OH NMe2
H H H H H H
OH OH OH
H2, Pd/C COOH
HO
NH2 NH2 Me
MeOH/ NH2 COOH
OH dioxane OH OH
OH O HO O O OH O HO O O OH O HO O O
Conover, L.H. 1955. U.S. Patent No. 2,699,054. H. Muxfeldt, et al. J. Am. Chem. Soc. 1979, 101, 689.

3
 Baran Lab The Tetracyclines D. W. Lin

Woodward's First Total Synthesis of a Biologically-Active Tetracycline, 6- O O

Demethyl-6-deoxytetracycline. Cl
Cl
MeO OMe
L.H. Conover, et al. J. Am. Chem. Soc. 1962, 84, 3222. (Initial communication) CO2Me
CO2Me (2 eq.)
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) then NaH (4 eq.),
then MeOH (1 eq.), OH
(i) O
rt --> 80 oC OMe O OH
OMe O
OMe O O O 45%
NaH
CO2Me intermolecular condensation outcompetes intramolecular!
DMF OMe OMe
(ii) O OMe Triton-B
dioxane-MeOH MeOH is essential to suppressing the kinetically
Cl Cl
favorable intramolecular condensation and
OMe OMe OMe O 50-70 oC permitting the intermolecular condensation with
NaH CO2Me
DMF 88% the oxalate prior to formation of the desired
55% tricycle. In the absence of MeOH, Woodward
observed formation of the intramolecular product OH
O O first, followed by condensation onto the oxalate O
CO2Me (i) AcOH/ OMe OH
CO2Me CO2Me
to form the five-membered ring shown:
H2SO4

(ii) H2SO4 Cl Cl O
CO2Me MeOH/CHCl3 CO2Me
CO2Me
OMe 44% OMe AcOH/HCl H CO2nBu

(i) H2, 200 psi (i) NaOH H2 O Mg(OMe)2

OH O
Pd/C CO2Me H2 O 90 oC toluene
AcOH, 30 oC 100 oC OMe O OH 73% OMe O OH reflux
52%
(ii) H2SO4 CO2Me (ii) I2, AcOH;
MeOH/CHCl3 then Cl2 in AcOH Cl CO2nBu Cl NMe2
93% OMe (iii) HF, neat H H
(i) NHMe2, -10 oC;
63% over 3 steps CO2nBu
(ii) NaBH4, -70 oC
Cl Cl
CO2H H2SO4 CO2Me
O 69% OH
MeOH/CHCl3 OMe O OH OMe O OH
reflux
66% The thermodynamically more favorable diastereomer is formed
OMe O OMe O exclusively in this step, with the carboxyamino substituent assuming
an equatorial position and thus establishing the cis relationship of
Chlorination blocks the para the bridgehead hydrogens. Ketone reduction is also stereoselective.
position, forcing condensation onto
the more hindered ortho position.

4
 Baran Lab The Tetracyclines D. W. Lin

NMe2
Cl NMe2 Cl NMe2 H H
H H H H NMe2
H H O
CO2nBu TsOH O 48% HBr
O
toluene 20 min CONH2
OH O
reflux CONHtBu
OH OH OH OH
OMe O OH 90% OMe O OH
OMe OH OH OH 15% bsm from A;
30% of A recovered
Zn dust
formic acid Cl NMe2 H2, Pd/C NMe2
H H Et3N H H
1 min O2
81% CO2H 91% OH
CeCl3
DMF-MeOH
glycine-NaOH buffer, CONH2
OH
OMe O OH pH = 10 OH O OH O
15 min Mixture of epimers at C4
(i) O
NMe2 NMe2
H H H H
OH This was a difficult step to optimize - Woodward himself noted dryly that "the case
CO2H O Cl at hand was by no means the smoothest we had encountered." Competitive
O hydroxylation at C11a was also observed, as well as rapid decomposition of the
(ii) O O
EtO2C product under prolonged reaction conditions, forcing Woodward and colleagues
OMe O OH OMe O OH NH
to halt the reaction prematurely.
EtO NHtBu t
Bu
Mg(OEt)
A No acylation of the enols by the
CaCl2 NMe2
chloroformate was observed.
6 H H
OH
NMe2
H H BuOH-H2O, pH = 8.5
NaH O ethanolamine buffer
reflux, 10 min CONH2
DMF/MeOH 6% over 2 steps, OH
O
120 oC EtO 10% recovered SM OH O OH O
15 min OMe O O NtBu 6-desmethyl-6-deoxytetracycline
O Thermodynamic equilibration
to the desired epimer.
NMe2 Observe the classic Woodward master
H H
O stroke. Despite the presence of four This was the first total synthesis of a tetracycline with all the requisite
enolates, we observe only one of two functionality for full antibiotic activity. Note, however, that this is not the total
plausible intramolecular condensation synthesis of a tetracycline natural product. Substituents at the C6 position are
CONHtBu events. The other event is impossible since missing.
the enolate double bond cannot rotate to
OMe OH OH OH bring the amide into position for cyclization.

5
 Baran Lab The Tetracyclines D. W. Lin

Shemyakin: The First Total Synthesis of a Tetracycline Natural Product

HO Me O HO Me NO2
A.I. Gurevich, et al. Tet. Lett. 1967, 8, 132. H Et3NH+ H H
M.N. Kolosov, S.A. Popravko, M.M. Shemyakin. Lieb. Ann. 1963, 668, 86. O 2N
OEt CO2Et
B.-M.G. Gaveby, J.C. Huffmann, P. Magus. J. Org. Chem. 1982, 47, 3779.
THF
Note that the Lieb. Ann. reference cites a number of obscure Russian journals.
The JOC reference, however, illustrates Shemyakin's approach to the tricyclic OBn O OH OBn O OH
precursor produced below.

O O Me NO2 Me NH2
H H H
Zn
BF3.OEt2 LiAl(OtBu)3H HCl CO2Et dust CO2Et
+ EtOH AcOH
86% 64%
H
OH O OH O OAc OBn OH O OBn OH O
OAc

O O (i) O (i) 0.1 N KOH, THF-H2O

H H MeMgBr
BnBr 6 eq O (ii) O
N O
K2CO3 74% OEt Me NPhth N
54% H
H H O OEt
OH OH OAc BnO OH OAc CO2Et
(ii) MeI, Ag2O O

OH OH
Me H Me H OBn OMe O
KOH/MeOH

85%
H
PCl5 in DMF, then
H
BnO OH OAc BnO OH OH O O
Me NPhth Me NPhth
H H
EtO NH2 OH
HO Me CO2H
H Mg(OEt)
Jones reagent EtO2C CONH2

60% OBn OMe O OBn OMe O

OBn O OH Notice Shemyakin adopting the Woodward approach to ring A.

6
 Baran Lab The Tetracyclines D. W. Lin

HO2C
Me NPhth O
H Na+
OH S Me HN
H
OH
DMSO
EtO2C CONH2
OBn OMe O CONH2
OBn OMe O OH

Me NMe2
H
OH
(i) HBr-AcOH O2 over Pt
(ii) MeI in THF Et3N
CONH2
THF
OH OH O OH rt, 8 hr
This intercepts a degradation product which had A.I. Gurevich, M.G. Karapetyan,
previously been elaborated into tetracycline. M.N. Kolosov. Khim. Prirodn.
Soedin., Akad. Nauk Uz.SSR
1966, 141.
Me NMe2 (i) O2, h
H
OH 3,4-benzopyrene (cat.)
benzene

CONH2
(ii) H2, Pd/C
OH
OH OH O O Mechanism? Answer on Slide 16.
M. Schach von Wittenau. J. Org. Chem. 1964, 29, 2746.

HO Me NMe2
H H
OH

CONH2
OH
OH O OH O
tetracycline

7
 Baran Lab The Tetracyclines D. W. Lin

Ph
Ph
Muxfeldt's Total Synthesis of 6-Desmethyl-6-deoxytetracycline
Cl N
O Cl N
H. Muxfeldt, W. Rogalski. J.Am. Chem. Soc. 1965, 87, 933. (Communication) O
H. Muxfeldt, E. Jacobs, K. Uhlig. Chem. Ber. 1962, 95, 2901. (Prep of precursors) HCl
O THF O
Cl Cl (i) NaOH
O O
(ii) pyrolysis, MeO
CO2Me CO2Me MeO O
Br NaOMe 160 oC
CO2Me
O O Ph
MeOH
CO2Me 85%
MeO2C CO2Me over three steps MeO2C Cl N
NHtBu O
OMe OMe taut.
NaH (2 eq.)
Cl Cl O
(i) H2SO4 THF-Et2O
CO2H CO2H MeOH MeO2C CONHtBu
H3PO4 35 oC, 24 hr
95% MeO O O
80 oC (ii) ethylene glycol
CO2H TsOH, benzene
quant. Ph Ph
OMe OMe O 91%
Cl N Cl N
O O
Cl Cl (i) MsCl O
CO2Me
pyridine O
LiAlH4 OH 97% CONHtBu CONHtBu
MeO2C MeO2C
benzene-Et2O (ii) NaCN MeO O O MeO O O
NaI (cat.)
O O 0 oC O O DMF-H2O
MeO MeO
94% 92% O O

Cl N Ph Cl HN Ph
Cl Cl O O O
taut.

CN Li(EtO) AlH CHO C6H5 N CO2H

3 H MeO2C CONHtBu CONHtBu
MeO2C
benzene-Et2O Pb(OAc)4 (cat.) MeO O O MeO O O
O O 0 oC O O Ac2O
MeO MeO
64%
Now the stage is set for the second cyclization in this magnificent transformation.
Only one equivalent of NaH used so far!

8
 Baran Lab The Tetracyclines D. W. Lin

Cl HN Ph Cl NHBz
Muxfeldt's Last Hurrah: Total Synthesis of Terramycin

O O H. Muxfeldt, et al. J. Am. Chem. Soc. 1968, 90, 6534.

NaH
H. Muxfeldt, et al. J. Am. Chem. Soc. 1979, 101, 689.

MeO2C CONHtBu MeO CONHtBu Terramycin is a much more difficult target than the prototypical tetracyclines
MeO O O MeO O O discussed previously - Woodward and Muxfeldt avoided many of the problems
O
outlined earlier with by targeting a structure without the troublesome C5 and C6
substituents, while Shemyakin targeted a tetracycline which did not have the
Cl NHBz Cl NHBz
C6 hydroxyl. Here Muxfeldt and colleagues (including a young Edwin Vedejs!)
O OH tackle those problems head-on! Sadly, this is reported in a posthumous
communication from the Muxfeldt laboratories.

CONHtBu CONHtBu
MeO O O O MeO O OH O O (i) Ac2O, H2SO4 O OAc (i) MeMgI,
H -65 oC
82% from the starting aldehyde 0 oC
isolated as mixture of epimers 83% 82%
at C4 (ii) 1-acetoxy- (ii) NaOH
Muxfeldt thus effects the construction of the A and B rings in a single step! The only butadiene 84%
H
problem, unfortunately, is the failure to control C4 stereochemistry. benzene, reflux AcO O
OH O
Cl NH2 60%
Me Me
OH
(i) Me3OBF4 H2, Pd/C, H2CO
OH OH (i) acetone, O O KClO3
Me H Me H
(ii) HBr/AcOH, Et3N, MeOH CuSO4 OsO4 (cat.)
CONH2
100 oC 84% 50 oC
OH O OH O
(ii) Ac2O, 89%
(i) deprotects the benzoyl amide; (ii) deprotects the remaining functional groups. NaOAc
H H
HO O 95% AcO O
NMe2 NMe2
Me Me Me Me
OH OH
O O O O
Me H Pb(OAc)4 Me H
CONH2 CONH2 OH O
OH 40 oC
OH O OH O OH O OH O
6-Demethyl-6-deoxytetracycline OH O
H H
AcO O AcO O
Here they intercept an intermediate from the Woodward synthesis. They also report Mixture of cis-diols
hydroxylation with O2 over platinum (Angew. Chem. Intl. Ed. Eng. 1962, 1, 157).

9
 Baran Lab The Tetracyclines D. W. Lin

Me Me Me Me Preparation of C:

O O DBU-AcOH, O O O O O NH3 O NH2 O

Me H piperidine Me H (i) O3
O (cat.) -50 oC MeO OMe MeOH MeO NH2
xylenes 33%
(ii) H2O
O reflux 68%
H CHO conc. HCl
52% over H O O O
AcO O two steps AcO O C
CHCl3
60% MeO NH2
(i) H
Me Me Me Me N Coupling of B and C: BuLi (1.0 eq), -78 oC,
2:1
O O CH2Cl2 : O O
then Me Me
Me H 0.5 N NaCO3 Me H Ph
in H2O 91% O O N
CHO CHO Me H
S
CHO 85% (ii) NaH, then
MOMCl
H O
AcO O O HO O 90%
Mixture of C5 epimers O O O
MOMO O
Me Me Me Me MeO NH2 THF, reflux
2h
O O O O
Me H silica gel, Me H
N deactivated Me Me S Me Me S
CHO
70-80%
O O HN Ph O O HN Ph
Me H H Me H H
OH OH
MOMO O MOMO O +
The thermodynamically more favorable epimer is obtained exclusively.
CONH2 MeO2C CONH2
Me Me H
S MOMO O OH O MOMO O O
O Ph Ph
O O N 27% Mixture of diastereomers at C4, C4a
N Me H
S
Once again, Muxfeldt employs his beautiful method for forming the A and B
Pb2(OH)(OAc)3 rings in a single step. And once again, there is little stereocontrol - all four
77% O possible epimers at C4 and C4a are formed in solution. Fortunately, the desired
diastereomer readily crystallizes. The reason for employing the thiazolanone
MOMO O
B
rather than the oxazolanone employed before will become clear shortly...

10
 Baran Lab The Tetracyclines D. W. Lin

Me Me S Me Me S S
9:1
O O HN Ph O O HN Ph OH OH HN Ph (i) MeI in THF OH OH NH3 Cl
Me H H AcOH : Me H H Me H H Me H H
OH H2 O OH
(ii) 0.17 N HCl
OH OH
in THF-H2O, 1.5 h
reflux,
CONH2 6 min CONH2 CONH2 CONH2
H 90% H OH OH
MOMO O OH O HO O OH O HO O OH O HO O OH O
No epimerization at C4 observed!
While the oxazolone substrate could also be carried to this step, the resulting
S
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.
OH OH HN Ph
Me H H
OH
12% OH OH NMe2
desired C12a Me H H
CONH2 Me2SO4 OH
hydroxylated
(i) P(OEt)3, NaH, O2 OH
HO O OH O product (i-Pr)2NEt
DMF-THF-H2O
15 min + 23% from CONH2
thioamide OH
HO O OH O
(ii) 0.01 N HCl Me Me S
in MeOH terramycin
rt, 1.5 h O O HN Ph
47%
Me H H
OH C11a
hydroxylated This concludes an elegant synthesis which assembles the A and B rings in a
byproduct single step. Unfortunately, Muxfeldt and colleagues never satisfactorily address
CONH2 the issues of controlling the C4 and C4a stereocenters, nor do they improve
OH
HO O OH O upon Woodward's solution to the C12a hydroxylation problem.

14% recovered SM
(i) hydroxylates the molecule; (ii) cleaves the acetonide. Unfortunately,
hydroxylation occurs principally at C11a. In a fortunate accident, however, it turned
out that the acetonide could not be cleaved unless the C12a hydroxyl was present.
Thus separation of the desired deprotected product from the undesired major
product was quite facile by polyamide chromatography.

11
 Baran Lab The Tetracyclines D. W. Lin

Stork: Controlling the C4, C4a Stereocenters O

O O
O OAllyl
G. Stork, et al. J. Am. Chem. Soc. 1996, 118, 5304. piperidine (11 eq)
O OAllyl AcOH (40 eq)
Me O
Here Gilbert Stork and colleagues take a completely different approach in order mol. sieves Me
to achieve stereocontrol at the C4 and C4a centers. CHO
benzene
O O OH 0 oC --> rt
Me
(i) MeMgBr 2.5 h
78% OH O
97% OH O
100% (ii) 45% overall yield from start
100% of the synthetic sequence!
OH O OH O OH O O
OBn
This sets the C6 stereocenter. Now watch Stork use this O
MeO2C OAllyl
stereocenter to bootstrap his way through the molecule...
N , then NMe2
OEt OEt Me2N O 4a
OEt O Me
O NaHMDS, then 5a H O
O n-Bu3SnH,
Br Me Me the above tricycle N Pd(PPh3)4
Br AIBN MeO2C
Br
N,N-dimethyl- 95% from
OBn
aniline, 90% the tricycle
OH O
98%
OH O OH O Here Stork exploits the stereochemistry of the tricycle to
direct conjugate addition to the more accessible face.
O O
Observe that the C5a and C4a stereocenters are now set.
, then
OH HO O O
Me OH NMe2
HS SH S TFA anhydride, then NMe2 Me H H
O 4a
Bu3SnOCH3 O
Me 5a 4a
BF3.OEt2 S the dithiane 5a H O N
0 oC, 15 min 92% 60 oC
N
88% OH O Stork postulates a ketene intermediate MeO2C
97% MeO2C MeO C
2
formed from the mixed anhydride. OH O OBn
OBn Mild reagent for
O O O O OH O lactone cleavage
(i) PhI(OTFA)2,
MeOH Me NMe2
O OAllyl O OAllyl H H This protects the C6 and
Me 92% Me
TMSCN O C10 hydroxyls, and sets
S
(ii) 5% aq. HCl CHO O N the stage for the
KCN remaining cyclization
S quant. 18-crown-6 MeO2C MeO C
2
steps.
Transketalization, followed OTMS OTMS OBn
OH O by hydrolysis to aldehyde. OH O

12
 Baran Lab The Tetracyclines D. W. Lin

KH Me NMe2 A Note on C12a Hydroxylation: This intercepts an intermediate which has

Me NMe2
H H (25 eq) H H been hydroxylated at the C12a position according to literature reports,
O O completing in principle the formal synthesis of tetracycline. However, Stork
O N -78 --> 0 oC, O N and colleagues were unable to successfully apply any of the C12a
3 h, then hydroxylation methods previously reported. The presence of the C4
MeO2C
MeO2C MeO C
2 0 --> 50 oC, BnO dimethylamino substituent seems to interfere with the hydroxylation. Clearly
OTMS OTMS BnO OTMS OTMS O
30 min a satisfactory solution to the C12a hydroxylation problem is still needed...

O NMe2 OH NMe2
Me H H Me H H
O O
N N

MeO
O O O O BnO OH O OH OH BnO
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 C11 ketone
resulted in formation of a BCD tricycle for which conditions to complete A ring
cyclization could not be found.

OH NMe2
H2 Me H H
OH
Pd black
94%
CONH2
OH O OH OH

12a-deoxytetracycline

13
 Baran Lab The Tetracyclines D. W. Lin

Tatsuta: Asymmetric Total Synthesis of Natural (-)-Tetracycline

CrO3, H2SO4
K. Tatsuta, et al. Chem. Lett. 2000, 647. OBn O NHCbz
H 0 oC, 10 min
OBn 85%
Here Tatsuta and colleagues not only produce an asymmetric total synthesis, but OTMS
they also take a very different approach to the synthetic problem, constructing OH
the A and B rings first and exploiting the carbohydrate chiral pool for starting BnO t t
Bu Bu H
materials. And as a bonus, they solve the C12a hydroxylation problem as well! NHCbz OH O OBn

(i) DMSO 170 oC

OTBS DCC, Py-TFA (i) HCl-MeOH Me 72%
OTBS
97% 93%
H 2C
O O (ii)
HO (ii) Ph3PCH3Br Me
BnO NO2
BuLi/THF, BnO
CbzHN CbzHN
OMe -78 oC --> rt NHCbz O Me OH NHCbz
OMe H H H
91% SeCN OBn OBn SOCl2
PBu3 (cat.) O Et3N
90% OMe
LDA, -40 oC, -30 oC
NO2 O OH OBn 15 min OMe OH O OH OBn 10 min
HO
(i) BnBr
80% 90%
BH3.THF, BaO/Ba(OH)2
Se 0 --> 45 oC; CH2 84%
O (i) BBr3
H 2C then H2O2, Me NHCbz Me NHBoc
O BnO (ii) HgCl2 H -78 oC H
NaOH/THF THF-H2O OBn OH
BnO 69% CbzHN 15 min
OMe 67%
CbzHN
OMe (ii) H2, Pd/C
Boc2O, Et3N
OMe OH O OH OBn 92% over OMe OH O OH OH
(i) MsCl, Et3N
BnO BnO two steps
0 oC, 15 min
CH2 O 82%
OH Me NHBoc
TMSCHN2 H
BnO BnO (ii) DBU, -30 oC i-Pr2NEt OH
CHO
CbzHN CbzHN quant.
OH 72%
In addition to eliminating to the enone, (ii) also
epimerizes to the thermodynamic diastereomer. OMe OH O OMe OH
Attempts to directly oxidize this 1,3-diol to the 1,3-dicarbonyl failed, requiring
the following detour of sequential alcohol oxidations.

14
 Baran Lab The Tetracyclines D. W. Lin

Me NHBoc Me NHBoc
H Br2 H
OH OH O
(Bu3Sn)2O
OH
Br
mol. sieves O N
N N
-78 oC, 15 min N N
OMe OH O OMe OH 85% OMe OH O O OMe
H
H
Me NHBoc
H
Dess-Martin O Zn, AcOH
periodinane 2 min (i) H3PO4 Me NMe2
Me NH2
15 min
Br H 100 oC,45 min H
OH OH
91% 68%
OMe OH O O OMe
(ii) H2CO,
CN CONH2
OH HCOOH OH
OMe OH O O 80 oC, 1 h OMe OH O O
Me NHBoc Me NHBoc
H Dess-Martin H 80%
O periodindane O

Me NMe2
15 min OH H
62% over BBr3 OH
O2, h
H
OMe OH O OMe OH two steps OMe OH O OMe
0 --> rt meso-tetraphenyl-
88% CONH2 porphyrin (cat.)
Cl
OH 10 min
B OH OH O O 75%
TsN NTs
O O Me NHBoc
H (i) H2NOH.HCl OOH NMe2
OH Me H
Ph Ph Et3N, 30 min
OH
O (ii) O
Et3N
-78 oC, 30 min OH CONH2
OMe OH O O N N N OH
60% N OH O O O

Here Tatsuta et al. employ DMDO to achieve the desired 80%over 2 steps
hydroxylation. They also achieve enantioselectivity by Here Tatstuta et al apply a protocol developed by Wassermann, Lu and
exploiting the chiral boron catalyst which Corey developed Mechanism? Scott for hydroxylating anhydrotetracyclines. Provide a mechanism for this
for enantioselective aldols and Diels-Alder reactions. Note reaction, and rationalize the stereospecific nature of this reaction.
the fantastic yield!
H. Wassermann, T.-J. Lu, A.I. Scott. J. Am. Chem. Soc. 1986, 108, 4237.

15
 Baran Lab The Tetracyclines D. W. Lin

Myers' Rapid Asymmetric Access to Analogs of Tetracycline

O O O
H O H M.G. Charest, C.D. Lerner, J.D. Brubaker, D.R. Siegel, A.G. Myers. Science 2005,
H
H H H 308, 395.
H
H
NMe2 NMe2 In an extraordinary report, Myers and colleagues present a highly efficient and
O O O O O O enantioselective method for accessing the tetracyclines.
H H H H
OH O H OH O H
mCPBA
H O H O A. eutrophus B9 EtOAc
N N 12a CO2H O CO2H
H H 83%
CO2H 75%, >95% ee OH OH
Wassermann, Lu and Scott invoke a formal ene reaction. The orbital OH OH
alignment requirements dictate that only the axial hydrogen can participate Notice how Myers begins with installation of the
in the reaction, inducing hydroperoxidation on the upper face of the This bacterial-catalyzed reaction
can be run on a 90 g scale! troublesome C12a hydroxyl group, and then
molecule and thus ensuring the proper stereochemistry at C6.
proceeds to build the molecule around it!

1. TMSCHN2 NMe2
OOH NMe2
Me H
OH NMe2 2. TBSOTf, Et3N THF, -78 oC
Me H H O
OH OH
H2/Pt 70% O NMe2 O N
TBSO CO2Me TBSO
O
CONH2 62% CONH2 OBn
OH OTBS N OTBS O
OH
OH O O O OH O OH O
Li 73%
(-)-tetracycline OBn

Me NMe2
1. LiOTf, Me H
There are many elegant features to this synthesis. Tatsuta and colleagues N O
mimic Stork's Diels-Alder approach to establishing stereochemistry, but employ toluene, 60 oC N O
it to define the troublesome C4a stereocenter immediately. They construct the B A N
2. TFA, CH2Cl2
central tetracycline scaffolding in just three steps from simple precursors. And 60% BnO HO
TBSO OH
they solve the C12a hydroxylation problem with a very mild oxidant in the OH TBSO O
OBn
presence of a chiral catalyst, and introduce the C6 hydroxyl stereospecifically TBSO O
21% over 7 steps
at a very late stage of the synthesis.
Here Myers closes the ring and sets the C4 amine stereochemistry. Myers
compares this key ring-closing step to a Sommelet-Hauser rearrangement, where
the amine initially undergoes an intramolecular SN-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!

16
 Baran Lab The Tetracyclines D. W. Lin

Now Myers takes his key intermediate A and converts it into two fragments: B, BnO2CO NMe2
which will go on to form C6-deoxy analogs of tetracycline, and C, which will go H
on to form analogs with the normal C6-oxygenation. O
N C
NMe2 NMe2
H H
O PPh3, DEAD; O O O
OBn
N N OTBS
then O2
HO With these fragments in hand, Myers now can install the C and D rings, and he
OH S NH2
OBn N OH OBn proceeds to do so in a fashion that allows for analogs of tetracycline with deep-
TBSO O H TBSO O
seated structural modifications to the D ring.
A NO2 74%
O

Me OBn
1. HCl, MeOH NMe2 Me O NMe2
H LDA, TMEDA,
2. IBX, DMSO THF, -78 oC; H H
3. TBSOTf, 2,6-lutidine O O
N B then C, N
CO2Ph
66% -78 oC -> 0 oC
OBn BocO 79% OBn
O O OH O OH O
OTBS OTBS
NMe2
H NMe2
H
O 1. CBr4, PPh3 Me OH NMe2
O
N 2. PhSH, Et3N H H
N 1. HF, MeCN OH
HO 2. H2, Pd, THF/MeOH
OH 87% PhS
OBn OH NH2
TBSO O OBn
TBSO O 90%
A OH
OH O OH O O

Me Me
Cl (-)-doxycycline
OH NMe2 1. BnO2CCl, DMAP 18 steps, 8.3%
1. Cl H
N 2. TBAF, HOAc
O 3. IBX, DMSO
S O N
O2 4. TBSOTf, Et3N

2. P(OMe)3, MeOH OH OBn

TBSO O 85%
70 oC
76%

17
 Baran Lab The Tetracyclines D. W. Lin

With this strategy, Myers and colleagues are able to synthesize a number of
remarkable analogs of tetracycline:

Me NMe2
H H
Me OH
B

CO2Ph NH2

OBoc OH
OH O OH O O
(-)-6-deoxytetracycline
NMe2
Me Me H H
B OH

N HN NH2
CO2Ph
OH
OBn O O OH O O

NMe2
H H
N Me N OH
B

NH2
CO2Ph
OH
O OH O O
NMe2
H H
CH2Br OH
B
NH2
CO2Ph
OH
O OH O O

NMe2
CH2Br H H
B OH

CO2Ph NH2
OMe OH
OMe O OH O O

18
 Baran Lab The Tetracyclines D. W. Lin

Addendum: Tetracycline Tidbits 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 spent over a decade tinkering with tetracycline, but never completed
a total synthesis of the molecule. Over the course of this work, however, he O O O O
discovered some interesting chemistry (naturally). R Me R
Me
PhSe SePh
O
Photocyclizations of acetals onto enones: Me R' Me R'

O O

h AcO AcO
H H H
H
ArCO2H
H D.H.R. Barton, D.J. Lester, S.V. Ley. J. Chem. Soc. Perkin Trans. I 1980, 2209.
O O O O
O O
Ph Ph
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]

O OH
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
H is the originality in the reactions and the reagents and any new principles
O O O that finally justify academic effort in synthesis. We are far away from the
O O O
Woodwardian dogma of completely planned synthesis."
Ph Ph

D.H.R. Barton, et al. J. Chem. Soc. Perkin Trans. I 1976, 503.

19