Total Synthesis of
7-Acetamidoquinoline-5,8-dione-2-carboxylic Acid
An Honors 499 Thesis
by
Michele Rae Etling
Mentor
Dr. Mohammad Behforouz
Ball State University
Muncie, IN
June 1, 1998
-
Graduation date: May 9, 1998
.
1
,i
.
.'
;
t"--, ,
I. Abstract
This thesis describes work done as part of an ongoing research project in the synthesis
oflavendamycin analogs, including various quinoline-5,8-diones. These smaller compounds
form two of the five rings found in the structure oflavendamycin, a potent antibiotic and
antitumor drug. The thrust of our research project is to find a compound with high antitumor
activity that also has low toxicity and sufficient solubility to make it suitable for clinical
applications.
-
The compound described in this thesis is a novel carboxylic acid derived from
compounds previously synthesized as part of Dr. Behforouz's research work. Although this
acid, due to its poor solubility, would not be suitable for practical applications, it is the hope
of our group that this compound may be used as an intermediate in producing other
compounds that would selectively enhance solubility and antitumor activity while decreasing
cytotoxicity.
Due to time constraints none of the acid derivatives (esters and amides) could be
prepared and purified prior to this writing. However, work on the project is ongoing by
several members within the research group.
2
Table of Contents
I. Abstract .......................................................................................... 1
II. Acknowledgements ......................................................................... 3
III. Background Information ................................................................. 4
IV. Synthesis ofQuinoline-5,8-diones .................................................. 6
V. Biological Activity .......................................................................... 9
VI. Synthetic Methods ......................................................................... 10
VII. Experimental. ................................................................................. 13
A. General Information ................................................................... 13
B. Solvent Purification ................................................................... 13
C. Procedures ................................................................................. 14
8-Hydroxy-2methyl-5,7-dinitroquinoline (2) ..................... 14
5,7-Diacetamido-2-methyl-8-acetoxyquinoline (3) ............ 14
7-Acetamido-2-methylquinoline-5,8-dione (4) .................. 15
7-Acetamido-2-formylquinoline-5,8-dione (5) .................. 15
7-Acetamidoquinoline-5,8-dione-2-carboxylic Acid (6) .... 16
N-Succinimide Ester .................................. 17
n-Butyl Amide ........................................... 17
Appendix A
NMR and IR Spectra .................................................. 18
Appendix B
Research Presentations ............................................... 23
Appendix C
Works Cited ............................................................... 24
3
--
II. Acknowledgements
I would like to thank first Dr. Mohammad Behforouz, a man whom I have come to
admire for his knowledge in many areas of life. He has taught me not only how to be a good
chemist, but to remember that the task at hand is not the most important thing in the world. I
am grateful for the opportunities he has given me to learn and to grow.
Mrs. Wen Cai has been a great help to all of us in the research group. I would like to
thank her for sharing her time, knowledge, and experience in helping me complete this work.
In the past two years she has provided me invaluable help in literally every step of this work.
I also thank the undergraduate and graduate students with whom I worked on a daily
basis. Our conversations eased my frustrations, gave me ideas for getting through the tough
spots, and helped the time pass - sometimes too quickly - while the TLC's were developing.
To the faculty and staff of the Ball State Chemistry Department, I owe you all many
thanks. Your willingness to share your time and expertise with all the students in the
department has made a huge difference. You are all teachers, in the greatest sense of the
word. Special thanks goes to Dr. Bruce Storhofffor letting me take up space in his lab. In
addition, I want to thank the Ball State Honors College for inspiring this work, and all my
professors in all of my classes who have challenged and inspired me over the past four years.
Much gratitude is owed to our funding sources for this ongoing project, including the
National Institutes of Health, the American Cancer Society, and Eli Lilly.
Finally, and most importantly, I offer my greatest thanks to my God and Savior, who
has made everything possible. Much thanks also for my parents, Richard and Karen Etling,
who have taught me and helped me so much through the years. I couldn't have done it
without you.
4
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III. Background Information
The quinoline-5,8-dione structure forms the AB ring portion of the antibiotic
lavendamycin. Lavendamycin, first isolated in 1981 at Bristol Laboratories, was found in the
fermentation broth of the bacterium Streptomyces lavendulae. 1 It has been shown to be quite
similar in both structure and biological activity to another compound known as streptonigrin,
previously isolated from S. .f/occulus. 2 ,3 Both lavendamycin and streptonigrin contain a
quinoline-5,8-dione moiety. Both have exhibited potent antitumor activity, however they also
have exhibited a high degree of cytotoxicity, making them unusable for clinical
applications. 4,5,6 Lavendamycin also has a low solubility in water, making it even more
impractical. l
o
o
'Z
10·
eOOH
¢o
o
OMe
Quinoline-5,8-clione
Although no single mechanism has been able to fully explain the cytotoxicity of these
and other quinones, several proposed mechanisms explain certain aspects of their biological
activity. One mechanism deals with the effects of quinone radicals on the electron transport
system in the mitochondria. 7 Several studies have observed the cleavage of single-stranded
DNA. 3,8,9,lO These studies correlate the biological activity to the reduction potential of the
quinones, especially to the formation of quinone free radicals. There is also evidence that
5
lavendamycin methyl ester and some of its analogs have an inhibitory effect on cdc25a
phosphatase, an essential component of cell cycle control. 11
Streptonigrin has been shown to have a much greater cytotoxic effect than many of
the quinoline-5,8-diones that have been tested? Lavendamycin, although less potent than
K
streptonigrin, has shown promising antitumor activity against ras transformed oncogenic
tumors. 11,12 Preliminary studies by our research group have indicated that the smaller
quinolinedione analogs are less potent than lavendamycin itself However, some of our
compounds have shown promise in reduced cytotoxicity to normal cells, while maintaining
some level of antitumor activity.
After the 1981 discovery of lavendamycin, several research groups started work on
methods to synthesize the entire pentacyclic structure. The first reported synthesis of
lavendamycin methyl ester resulted from work by Kende et al at the University of
Rochester. 14, 15 Their method involved a Friedlander condensation to form the AB ring
portion, followed by a Bischler-Napieralski cyclodehydration to produce the pentacyclic
product. Other methods involved a Pictet-Spengler condensation to join a quinoline analog
and f3-methyl tryptophan, forming the main skeleton. These methods involved many steps
with overall yields of 1% or less.
The first short, practical synthesis oflavendamycin methyl ester was devised by the
Behforouz group at Ball State University and reported in 1993. 16 This five-step synthesis
used a novel azadiene in a Diels-Alder reaction to form the AB ring system, followed by an
oxidation and a Pictet-Spengler condensation to give the pentacyclic product, with an overall
yield of33%. In 1996, Behforouz's group reported a more practical five-step synthesis that
increased the overall yield of lavendamycin to nearly 40%.12,17
6
While research is ongoing to study the structure-activity relationships of various
lavendamycin analogs, the Behforouz group is also looking at the AB ring system, the
quinoline-5,8-diones. While the work in lavendamycin analogs continues, many of the
bicyclic analogs have been synthesized to test for biological activity. Even though most of
the quinolinediones tested so far are less potent than lavendamycins, early indications show
that, with appropriate side group additions and modifications, it may be possible to produce a
quinoline-5,8-dione derivative that may have practical clinical applications. 1l , 13
Based on the structure-activity relationships in lavendamycin analogs, several
predictions have been made concerning the probable activity of different quinolinediones.
By adding or altering side groups at strategic locations, we hope to increase antitumor
activities while maintaining low cytotoxicity toward normal cells. In addition, we predict
that certain polar side groups will increase the solubility of the compounds, making them
more usable in clinical applications. 1l
Among those modifications predicted to increase the usefulness of our compounds
are the introduction of various esters, amides, and other acid derivatives at the C-2 position.
The C-2 carboxylic acid, a necessary functional group for these derivatives, has not
previously been synthesized in this class of compounds.
IV. Synthesis of Quinoline-5,8-diones
The synthesis of the bicyclic quinoline-5,8-dione skeleton was worked out as part of
the ongoing effort to synthesize numerous analogs of lavendamycin. Kende and Ebetino of
the University of Rochester were the first to publish a method for synthesis in 1984, using a
Friedlander condensation to produce the AB ring system as part of the synthesis of
7
lavendamycin methyl ester. 14 Boger obtained a quinoline-5,8-dione in 1985, using 8hydroxyquinoline as a starting compound. 19 In 1993, Behforouz's group reported the
synthesis of a 7-acetaminoquinoline-5,8-dione as part of a concise lavendamycin methyl ester
synthesis. 16 This method used a Diels-Alder condensation to of a bromo quinone and a novel
azadiene to form the quinolinedione AB rings (see Scheme 1). In two steps, the Behforouz
procedure produced a quinoline-5,8-dione with over 45% yield. This dione was then
functionalized to an aldehyde to facilitate formation of the pentacyc1e.
Scheme 1
'
+-
SiONH 2
,
+
~CHCI
r.t.
2
~
2 ..
0
~+
O-Si+
I
--
heat
se0 2
CH 3
0
ACHNYsr
0
..
AcHN
+
0
0
CSH5C1
n
0
..
AcHN
HO
0
In 1996, the Behforouz group reported a second short synthesis of lavendamycin
methyl ester with a 40% overall yield. 17 This method uses 8-hydroxyquinaldine (8-hydroxy2-methylquinoline) as the starting material for the AB ring system (see Scheme 3). This
procedure is much simpler in practice than the 1993 method, and gives comparable yields for
the quinolinedione. It is this procedure which was employed in this thesis to produce the
compounds described herein.
As part of the synthesis of compounds in the quinoline-5,8-dione class, we wanted to
make a group of esters and ami des at the 2-C position. This required, as a preparatory step,
-
the functionalization of this side group to an acid. Behforouz's group had formed an
8
aldehyde at this position, but had not been able to oxidize the group further. 16. 17 The
problem we had was that, in the strong acidic or basic environments required by most strong
oxidizing agents, the acylamino group in the 7-C position was easily hydrolyzed, giving an
amine product. Several attempts were made to find an oxidizing agent which was strong
enough to fully functionalize the acid at C2, but with a mild environment to protect the
existing amide at C7.
As it turned out, our group actually found two methods for making the acid (refer to
Scheme 2). The first procedure was determined by Darric Baty and involved simply
increasing the running time and amount of oxidizing agent in the existing procedure for
making the aldehyde?O This one-step reaction oxidized the methyl group directly, instead of
isolating the aldehyde. Unfortunately, though the presence of the desired acid was
confirmed, the product was difficult to purify and the reaction in general was cumbersome.
Scheme 2
o
5e02 reflux. 2-4 days
RHN
..
RHN
o
5eo~
16-20 hrs
RHN
C
II
.....OH
o
o
~.rt
CHO
90 min
The second method determined to form the acid is described in this thesis. It was
inspired by work from Banerjee et at and involved the use of sodium perborate to further
oxidize the aldehyde to a carboxylic acid. 21 This turned out to be a relatively simple reaction,
giving a good yield and a relatively pure acid product as a precipitate.
9
Once the acid was obtained, we continued our research to find a method of forming
different acid derivatives at this position. We intended to make the succinimide ester as an
intermediate to other esters and amides?2 Unfortunately, the polar compound was not
soluble in many organic solvents, and it was difficult to find a suitable solvent which would
not interfere with the reaction. DMF was the only suitable solvent available, but its very high
boiling point made the isolation ofthe product difficult. It was decided to continue with the
next step without the isolation of the succinimide ester intermediate, adding an amine directly
to the reaction mixture to form the amide. Due to constraints oftime, we were unable to
completely purify the product from this reaction, but we believe the compound does contain
the desired amide, with an amino group at the C-7 position, as shown by IH NMR.
v.
Biological Activity
The development of concise, high-yield methods for the production oflavendamyacin
and its analogs has made it possible to study the relationship of chemical structures in these
compounds to their biological activity. For several years, the Behforouz group has been
conducting ongoing structure-activity relationship studies on lavendamycin and a variety of
its analogs. These and studies by other groups have indicated that it may be possible to
achieve a potent antitumor drug using derivatives of the AB ring system, the quinoline-5,8diones.2, 11
-
10
-
VI. Synthetic Methods
The synthesis of the bicyc1ic quinoline-5,8-dione skeleton was worked out as part of
the ongoing effort to synthesize numerous analogs oflavendamycin. Behforouz's 1996
procedure for the synthesis of lavendamycin methyl ester involved making the AB ring
system from a quinoline derivative and functionalizing the aldehyde before undergoing a
condensation with a tryptophan to form the pentacyc1ic product. 17
The quinoline-5,8-dione was prepared according to the following process (refer to
Scheme 3). From Aldrich, 8-hydroxyquinaldine was purchased. A nitration was performed
in a mixture of concentrated nitric and sulfuric acids, yielding a dinitro product 2. As the
starting compound had an activating hydroxyl group in the 8 position, the nitro groups were
--
added to the ortho- and para- positions at 7 and 5. The reaction is quite exothermic, and
produces large quantities of NO gas; the reaction was kept under a hood, and in an ice bath to
keep the reaction temperature below 50°C.
Scheme 3
H2 /Pd-C 5% ..
cone HCI
Acetic Anhydride ..
Na 2S0 4 ' NaOAc
CH3
CH3
OH
NHAe
..
glacial acetic acid
CH 3
AeHN
OAe
3
AeHN
CH 3
o
AeHN
4
Following the nitration, 2 was reduced to give the amino product. The reaction
-
utilized a Parr hydrogenator at an initial H2 pressure of 43 psi over 24 hours, with palladiumon-charcoal as a catalyst. Since the electron donating effects of the amino groups on the
11
aromatic ring made the resulting amino compound sensitive to air oxidation, hydrochloric
acid was included in the hydrogenation mixture to protect compound by converting the
reduced amino groups to ammonium chloride salts. This salt was not isolated, but was
placed directly into solution with sodium sulfite, an antioxidant, and sodium acetate, which
acted as a base. The slow addition of acetic anhydride yielded crystals of the diacetamido 3.
This substitution reaction can be explained by the nucleophilic attack of the amino groups on
the carbonyl carbons of the anhydride.
Next, the diacetamido compound 3 was oxidized with potassium dichromate in a
solution of glacial acetic acid to give the quinolinedione 4. The aqueous reaction mixture
was extracted with dichloromethane. The organic layers were neutralized with 5% sodium
bicarbonate, dried with magnesium sulfate, and rotary evaporated to yield the yellow dione.
The 7-acetamido-2-methylquinoline-5,8-dione 4 was then oxidized to the aldehyde 5.
The reaction occurred in a wet solution of 1,4-dioxane under argon with selenium dioxide as
the oxidizing agent. The mechanism of this reaction is such that the selenium dioxide and
water react first, making a reactive selenic acid, which then selectively oxidizes only the
methyl group of the quinone to an aldehyde.
Two methods were employed to obtain the carboxylic acid 6, one by further oxidation
of the aldehyde in 5, the other by oxidizing the methyl group of 4 directly (see Scheme 2).
The first method turned out to be the most practical. Aldehyde 5 was treated with sodium
perborate in glacial acetic acid, yielding a precipitate of the 2-carboxylic acid 6 (refer to
Scheme 4).
The low solubility of the acid in suitable solvents made the formation of esters and
amides from 6 difficult. The acid was combined with N-hydroxysuccinimide in dry DMF,
12
and dicyc1ohexy1carbodiimide was used to facilitate the dehydration to the ester 7. We were
unable to successfully isolate the product from the DMF, so the reaction proceeded with the
direct addition of dry n-butylamine. We had hoped to obtain the amide product 8a, but we
Scheme 4
0
NaB0 4 , rt
glacial acetic aCid-
CH
II
0
AcHN
0
AcHN
0
6
C
II
0
+HO-N~
... OH
DMF, cold
7
urea
0
-
0
0
I~
h
N
AcHN
o
""""
Dee
7
o
~
C
___ /
............, ............,
... N~
II
8a, R=Ac 0
8b,R=H
believe that the basic conditions introduced with the addition of the amine caused the
aminolysis of the 7-acetamido group, yielding 8b. Pure product has not yet been obtained,
despite multiple attempts at column chromatography and other methods.
Due to time constraints, the author of this thesis was unable to obtain any purified
derivatives from the acid. However, work on this project is ongoing, and these derivatives
will likely be obtained in the near future.
13
-
VII. Experimental
A. General Information
Reagents: 8-hydroxyquinaldine, 5% palladium-on-charcoal, selenium dioxide, Nhydroxysuccinimide, and N-dicyclohexylcarbodiimide were purchased from Aldrich.
Sodium perborate was purchased from Mallinckrodt. All other reagents and solvents
were purchased from Fisher.
Solvents: All solvents used were reagent grade, with two exceptions. DMF was first dried
over molecular sieves, and 1,4-dioxane was dried according to method below.
Melting Points: All melting points were determined using a Thomas-Hoover capillary
melting point apparatus and are uncorrected.
Nuclear Magnetic Resonance Spectra: IHNMR Spectra were recorded on a Varian
Gemini 200 Spectrometer. Data for compounds 1-5 from thesis of A S. Rose;
spectra were taken in CDCh. Due to solubility, spectra of6, 8 taken in DMSO.
Infrared Spectra: FT-IR spectra were recorded on a Perkin-Elmer Spectrum 1000 FT-IR
Spectrometer with compounds suspended in a KEr mull wafer.
B. Solvent Purification
1,4-dioxane had to be purified due to its tendency to polymerize. The solvent was
refluxed over a large amount ofKOH, then decanted to remove the residue that formed. It
was then dried by refluxing with sodium spheres for 1-2 hours (until the spheres appeared
mirror-like), and then adding benzophenone to the mixture. A deep blue color from the
-
benzophenone indicated dryness, and the solvent was then distilled.
14
-
c.
Procedures
Preparation of 8-Hydroxy-2-methyl-5,7-dinitroquinoline (2)
In a 500-mL Erlenmeyer flask, concentrated sulfuric (30 mL) and concentrated nitric
(70 mL) acids were combined while stirring on an ice bath. While this acidic mixture was
kept at 20-30°C, 8-hydroxyquinaldine (1, 10.Og, 0.063mol) was added in small amounts over
15 minutes. Once the addition was completed, the reaction was allowed to stir an additional
3 hours. The mixture was then poured into a l-L beaker containing 600 mL ice cold water.
A bright yellow solid formed instantly and was then filtered off. The solid was allowed to
dry on the filter paper, then removed and mixed with ethanol (95%,200 mL), refiltered, and
washed with diethyl ether. When dry, the yield was 8.30g (53%) of product 2. mp.293296°C. lR NMR (CDCh): 0 9.65 (lR, d, J=9.lHz, C-4H), 9.20 (lR, s, C-6H), 8.13 (lR, d,
-
.
J=9.lHz, 3-CH), 2.93 (3R,
S,
C-2CH3) .
Preparation of 5,7-Diacetamido-2-methyl-8-acetoxyquinoline (3)
In a thick-walled 500 mL hydrogenation flask, concentrated hydrochloric acid (26
mL) and water (200 mL) were combined with finely ground 8-hydroxy-2-methyl-5,7dinitroquinoline (2; l2.00g, 0.048 mol). To the mixture, 4g of 5% palladium-on-charcoal
was added, and the flask was quickly placed on a Parr hydrogenator and shaken overnight, at
H2 pressure starting at 43 psi and falling to 22 psi. The next day, the flask was removed from
the hydrogenator and the reddish mixture was quickly filtered to remove the palladium-ocharcoal. The cake was rinsed with water (2 x 25 mL), and the filtrate was moved to a 500
mL round-bottom flask. To the dark red solution, sodium sulfite (8g) and sodium acetate
-
(lOg) were added and stirred magnetically, then 40 mL acetic anhydride was slowly poured
in. The flask was then placed on a rotary evaporator, and the solution was evaporated until
15
-
the first few crystals appeared. The flask was then cooled on an ice bath and stirred rapidly
while 40 rnL of acetic anhydride was added dropwise. The crystals that formed were then
filtered off and rinsed with water to give a pale yellow to white solid. The filtrate was once
again rotary evaporated to about
v.. its previous volume.
The flask was cooled and stirred
rapidly while adding an additional 80 rnL of acetic anhydride. The remaining precipitate was
filtered, rinsed with water, and combined with the first filtrate for drying, yielding 11.69g of
product 3 (77%).
Preparation of 7-Acetamido-2-methylquinoline-5,8-dione (4)
In alL Erlenmeyer flask equipped with a magnetic stir bar, mixed 240 mL glacial
acetic acid with 5,7-diacetamido-2-methyl-8-acetoxyquiniline (3, 6.3g, 0.020 mol). To this
-
suspension, a mixture of potassium dichromate (17.6 g, .06 mol) in water (200 rnL) was
added. The mixture stirred at room temperature for 24 hours, and was then added to a 2 L
separatory funnel containing 900 rnL of water. The solution was extracted with
dichloromethane (5 x 200 mL); the combined organic extracts were then washed with 5%
aqueous sodium bicarbonate (3 x 300 rnL) and dried over magnesium sulfate. After filtering,
the organic extracts were rotary-evaporated to dryness, then thoroughly dried on a vacuum
pump to yield 2.34g (51%) of yellow product 4. mp.216-219°C. lRNMR (CDCh): 08.41
(IR, s, C-NH), 8.32 (IR, d, J=8.1Rz, C-4H), 7.92 (IR, s, C-6H), 7.57 (1H, d, J=8.1Hz, C3H), 2.78 (3R, s, C-2CH3), 2.33 (3H, s, NHCOC~).
Preparation of 7-Acetamido-2-formylquinoline-5,8-dione (5)
In a 100 mL flask equipped with column, argon flow, and magnetic stir bar, placed 7acetamido-2-methylquinoline-5,8-dione (4, 1. 15g, 4.7 mmol) with 1.08g selenium dioxide,
17.5 mL purified, dried l,4-dioxane, and 0.625 mL of water. The mixture was heated slowly
16
-
to reflux over a two hour period, then was allowed to reflux overnight at 115°C. After 16
hours, the progress of the reaction was checked by TLC (1: 1 CH2Ch I EtOAc), and hourly
after that until reaction was complete (17-20 hours total). When reaction was complete,
added an additional 15 mL purified dioxane and continued refluxing for 15 minutes. The
mixture was vacuum filtered while hot to remove the selenium. The metal precipitate was
washed with 50 mL boiling dichloromethane, and the filtrates saved. The filter cake was
removed and repeatedly washed with hot dichloromethane to obtain as much product as
possible. The filtrates were then combined and rotary evaporated to dryness. The yellow-tan
solid was dried on a vacuum pump to yield 0.84 g (69%) of aldehyde 5. mp.220-223°C. IH
NMR (CDCh): 010.31 (1H, s, ArCHO), 8.64 (1H, d, J=7.7Hz, C-4H), 8.46 (lH, s, C-7NH),
8.34 (lH, d, J=7.7Hz, C-3H), 8.07 (lH, s, C-6H), 2.37 (3H,
-
S,
COCH3).
Preparation of 7-acetamidoquinoline-5,8-dione-2-carboxylic acid (6).
In a 50-mL pear- or round bottom flask equipped with a magnetic stir bar, 5 mL of
glacial acetic acid was heated to 90°C on an oil bath. To the warm acid, the aldehyde 5
(488mg, 2 mmol) was added and allowed to stir 15 minutes. Sodium perborate (1.54g, 10
mmol) was added to the suspension in small portions over 30 minutes, and an additional5mL
of acetic acid was added as the mixture thickened. The mixture was allowed to stir for an
hour, then 10 mL of water was added and allowed to stir a few minutes. The flask was then
removed from the oil bath and the reaction was vacuum filtered to obtain the yellow
precipitate. The filtrate was then rotary evaporated to about 113 original volume, then
acidified (pH 2) with sulfuric acid. The crystals which formed were filtered, added to the
-
first filter cake, and dried to give 300mg (57.7%) of acid 6. mp.248-250°C. IHNMR
17
-
C-3H), 7.78 (IH, s, C-6B), 2.27 (3H, s, COCHJ). FT-IR{KBr): 3278, 3221,1764,1682,
1646, 1525, 1357, 1307, 1196 cm- I .
Preparation of N-succinimido 7-acetamidoquinoline-5,8-dione-2-carboxylate (7)
In a dry 50-rnL round bottom flask equipped with a stir bar and an argon balloon,
placed 7-acetamidoquinoline-5,8-dione-2-carboxylic acid (6; 130mg, 0.5 mmol) and 5 rnL of
DMF. The mixture was heated at 70°C and stirred until the solution cleared, using additional
DMF as needed (maximum of 10 rnL). The solution was cooled under argon flow and placed
in an ice bath. N-hydroxysuccinimide (60mg) and 110 mg dicyclohexylcarbodiimide were
added, and the reaction was allowed to stir overnight. The stirrer was then stopped and the
flask was cooled in a fresh ice bath to allow the precipitate to settle out. The mixture was
-
then vacuum filtered to remove the urea. Attempts to isolate the succinimide ester 7 by
removing the DMF gave very low yield, so the reaction proceeded directly to the next step.
Preparation of N-Butyl 7-acetamidoquinoline-5,8-dione-2-carboxamide (8)
To the filtrate solution containing 7, dried, distilled n-butylamine (54 J,JL, .55 mmol)
was added using a Hamilton micropipette. The addition of the base caused an aminolysis at
the 7-acetamido group, indicated by the red color of the final product. The flask was placed
on a vacuum-distillation unit to allow room-temperature removal of the DMF. The dried
residue contained a mixture which was difficult to purify. The major compound appears to
be 8b, the 7-amino product. Column separation did not fully purify, and NMR is very
complex. IR (KBr): 3288,2959,2931,2870, 1717, 1676, 1608, 1581, 1527, 1257 cm- I .
18
Appendix A:
NMR and IR Spectra
Index of Spectra
7 -Acetamidoquinoline-5 ,8-dione-2-carboxylic Acid (6)
IH NMR (DMSO) .................................................................................... 19
FT-IR (KBr mull) , .................................................................................... 20
7-Aminoquinoline-5,8-dione-2-carboxyl n-Butyl Amide (8b)
IHNMR (DMSO) .................................................................................... 21
FT-IR (KBr mull) ..................................................................................... 22
.-
--
.-
L
:c
w()
)=0
:CZ
o
I
o
I
o=()
'0:c
-
-
20
72.4l
70
I
68
FT-IR (KBr mull)
66
64
o
62
H
60
58
o
56
6
%T 54
52
50
48
46
44
421
40
38
1
I
37.1+1--------------~--------------~----------~--,_--------------,_----------~
4000.0
3000
1500
2000
1000
600.0
cm-l
_ _ c:\pel_data\etling\acid1.sp
(
(
(
.-
....
0
:i
I
IV
I
Z
0
0
~
"......,
tJ
~
Ct:l
I
0=0
0
'-'
I
\
ZI
<
-
-
N
22
75.4
75
74
FT-IR (KBr mull)
73
I
72
71~
H
H2N
70
0
8b
69
%T 68
67
66
65
64
63
62
61.3
4000.0
3000
2000
1500
1000
600.0
cm-l
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23
Appendix B:
Research Presentations
The author has presented all or part of the work described in this thesis on two
occasions: once for the Ball State Chemistry Department, and once at the 113th Annual
Indiana Academy of Sciences meeting on October 31, 1997 at Saint Joseph's College .
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.
24
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Appendix c:
Works Cited
l. Doyle, T. W.; Balitz, D. M.; Grulich, R E.; Nettleton, D. E.; Gould, S. 1.; Tann, C. H.;
Meows, A E. "Structure Detennination ofLavendamycin: A New Antibiotic From
Streptomyces lavendulae." Tetrahedron Letters. 1981,22,4595
2. Boger, D. L.; Yasuda, M.; Mitscher, L.; Drake, S. D.; Kitos, P. A "Streptonigrin and
Lavendamycin Partial Structures. Probes for the Minimum, Potent Phannacophore of
Streptonigrin, Lavendamycin, and Synthetic Quinoline-5,8-diones." Journal of
Medicinal Chemistry. 1987,30, 1918 .
3. Lown,1. W.; Sim, S.-K "Studies Related to Antitumor Antibiotics. Part VIII. Cleavage
of DNA by Streptonigrin Analogues and Relationship to Antineoplastic Activity."
Canadian Journal ofBiochemistry. 1976,54,446.
4. Balitz, D. M.; Bush, 1. A; Bradner, W. T.; Doyle, T. W.; O'Herron, F. A; Nettleton, D.
E. "Isolation ofLavendamycin: A New Antibiotic from Streptomyces lavendulae." The
Journal ofAntibiotics. 1982,35,259.
-
5. Hackethal, C. A; Golbey, R B.; Tan, C. T. C.; Kamofsky, D. A; Burchenal, J. H.
"Clinical Observation on the Effects of Streptonigrin on Patients with Neoplastic
Disease." Antibiot. Chemother. 1961, 11, 178.
6. Wilson, W. L.; Labra, C.; Barrist, E. "Preliminary Observation on the Use of
Streptonigrin as an antitumor Agent in Human Beings." Antibiot. Chemother. 1961,11,
147.
7.
Bachur, N. R.; Gordon, S. L; Gee, M. V. "A General Mechanism for Microsomal
Activation of Quinone Anticancer Agents to Free Radicals." Cancer Research. 1978,
38, 1745.
8.
Shaikh,l A; Johnson, F.; GroHman, A P. "Streptonigrin 1: Structure-Activity
Relationships among Simple Bicyclic Analogues. Route Dependence of DNA
Degradation on Quinone Reduction Potential." Journal ofMedicinal Chemistry. 1987,
29,1329.
9. Cone, R; Hasan, S. K; Lown, 1. W.; Morgan, A R "The Mechanism of the
Degradation of DNA by Streptonigrin." Canadian Journal ofBiochemistry. 1976,54,
219.
-
10. Lown, 1. \V.; Sim, S.-K; Chen, H.-H. "Hydroxyl Radical Production by Free and DNABound Aminoquinone Antibiotics and its Role in DNA Degradation. Electron Spin
Resonance Detection of Hydroxyl Radicals and by Spin Trapping." Canadian Journal of
Biochemistry. 1978, 56, 1042.
11. Behforouz, M. Grant Proposal to the National Institutes of Health, 1997, unpublished
25
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12. Behforouz, M.; Merriman, R L. "Lavendamycin Analogs and Methods of Making and
Using Them." United States Patent 5,525,611. June 11, 1996.
13. Behforouz, M.; Merriman, R L. "Methods of Using Lavendamycin Analogs." United
States Patent 5,646,150. July 8, 1997.
14. Kende, AS.; Ebetino, F. H. "The Regiospecific Total Synthesis ofLavendamycin
Methyl Ester." Tetrahedron Letters. 1984,25,923.
15. Kende, AS.; Ebetino, F. H; Battista, R; Lorah, D. P.; Lodge, E. "New Tactics in
Heterocyclic Synthesis." Heterocycle. 1984,21,91.
16. Behforouz, M.; Gu, Z.; Cai, W.; Hom, M. A; Ahmadian, M. "A Highly Concise
Synthesis ofLavendamycin Methyl Ester." The Journal a/OrganiC Chemistry. 1993,
58, 7089.
17. Behforouz, M.; Haddad, J.; Cai, W. Arnold, M. A; Mohammadi, F; Sousa, A C.; Hom,
M. A "Highly Efficient and Practical Syntheses ofLavendamycin Methyl Ester and
Related Novel Quinolindiones." The Journal a/Organic Chemistry. 1996,61,6552.
-
18. Boger, D. L.; Duff, S. R; Panek, M.; Yasuda, M. "Inverse Electron Demand Diels-Alder
Reactions of Heterocyclic Azadienes. Studies on the Total Synthesis ofLavendamycin:
Investigative Studies on the Preparation of the DCE p-Carboline Ring System and
ABQuinoline-5,8-quinone Ring System." The Journal a/Organic Chemistry. 1985,50,
5782.
19. Behforouz, M.; Haddad, 1; Cai, W.; Gu, Z. "Chemistry of Quinoline-5,8-diones." The
Journal a/Organic Chemistry. 1998,63,343.
20. Baty, Darric. Lab notes, unpublished.
21. Banerjee, A; Hazra, B.; Bhattacharya, A; Banerjee, S.; Banerjee G. C.; Sengupta, S.
"Novel Applications of Sodium Perborate to the Oxidation of Aromatic Aldehydes, aHydroxycarboxylic Acids, 1,2-Diketones, a-Hydroxyketones, 1,2-Diols and Some
Unsaturated Compounds." Synthesis. 1989, 765.
22. Anderson, G. W.; Zimmerman, 1 E.; Callahan, F. M. "The use of Esters ofNHydroxysuccinimide in Peptide Synthesis." . 1964,8x, 1839.
23. Rose, A S. "Total Synthesis of7-N-Acetyldemethyllavendamycin sec-Butyl Ester and 7N-Acetyldemethyllavendamycin sec-Butyl Ester." 1997. Honors 499 Thesis.