Note: Descriptions are shown in the official language in which they were submitted.
WO 91/1605~ 2 ~ ~ 8 7 7 ~ PCI'/US91~02005
2 ~5H) -FlJ~AN0NE:8 ~UB81~TU~5!ED ~N T~ 5 AND OR IN T~}~ 4 ' -
PO8I~ION, A8 ANT~ TORY AGENTR
BAC~GRO~ND OF T~ ION
1. Field of the InventiQn
The present invention is directed to novel 2(5H)-
furanones substituted in the S position and or in the 4
position, which compounds are active as anti-inflammatory
agents. The present invention is also directed to
pharmaceutical compositions which comprise one or more of
10 the novel compounds of the invention, to the methods of
using these pharmaceutical compositions, and to the
chemical processes of making the novel compounds.
2. Brief Description of the Prior Art
Manoali~e is a compound isolated from a marine sponge
15 ~E. D. de Silva et al., ~etrahedron Letters 21:1611-1614
(1980)] which has anti-inflammatory, immunosuppressive and
analgesic properties. Manoalide (Co~poun~ 1) the
structure of which is shown below, includes a 5-hydroxy-
2(5H)-furanone moiety, attached in the 4-position of the
20 furanone ring to the rest of the molecule. Certain
analogs of manolide, such as -co-~no~ (Co~poun~ 2)
and dehydro-s-co-~a~o~lide (Compou~d 3) also have anti-
inflammatory activity. For further description of the
biological activity of ~anoalide and some of its
25 derivatives reference is made to United States Patent Nos.
4,447,445, 4,786,651, 4,789,749 and to European Patent
Application No. 0 133 376 (published on February 20,
1985).
. ~ - ~ ;- . .
WO91/16055 PCT/USsl/02~s _
2~g77~
S ~b~H
manoalldQ
dehydro-seco- ~OH
Compound 1 ~
Compound 3
~;~OH
seco manoalide p~--OH
0
Compound 2
Synthetic analogs of manoalide, particularly analogs
having various sub~tituents on the furanone moiety of
manoalide, are describ~d in s~veral applications for
United States Lett~r~ Patent by the same inventor as in
the pre~ent application, the following of which have been
allowed and ara expected to i~u~ as United States Letters
Patent:
Serial No. 259,225 filed on October 18, 1988;
Serial No. 281,126 ~iled on Dec~mb~r 7, 1988.
Publi~h~d Europo~n Patent Applicatlon No. 0 295 056 -
disclose~ 4 ~ub~tituted 5-hydroxy-2(SH)-furanones having
, . ... . ,.. .~ ,. .. - . . ., - ~ - ,. . , . . .,: . , -
WO91/16055 2 ~ 7 ~ 7 7 ~ PCT/USg1/02~5
anti-inflammatory, immunosuppressive and anti-
proliferative activity where the substituents in the 4
position are a variety 1-hydroxyalkyl, l-acyloxy-alkyl and
l-carbamoyloxy-alkyl groups.
United States Patent No. 4,855,320 discloses 5-
arylalkyl-4-alkoxy-2(5H)-furanones as anti-convulsive and
anti-epileptic agents.
Published European Patent Application No. 0 209 274
discloses 4-alkyl-5-hydroxy-2~5H)-furanones as anti-
10 inflammatory and anti-allergy agents.
Chemical Abstracts Volume 107 236559t (1987) .
discloses 4-acyloxy 5-hydroxy-2(5H)-furanones.
~MMARY OP T~E INV~NT~ON
The present invention covers compounds of ~or~ula 1,
and compound-~ of Formula 2
R~ ~ R~ R~ ~
For~ula 1 Yor~ul~ 2
- .. . . -. , : ,
WO9tJ1605~ 2 ~ ~ 8 ~ ~ ~ PCT/US91/02~5-_
~n which Rl is H, alkyl of l to 20 carbons, alkylene
having one or more double bonds, alkyne having one or more
triple bonds, arylalkyl, arylalkylene having one or more
double bonds or arylalkyne having one or more triple
bonds:
R2 is H, alkyl of l to 20 carbons, alkylene having one or
more double bonds, alkyne having one or more triple bonds,
arylalkyl, arylalkylene having one or more double bonds or
arylalkyne having one or more triple bonds;
10 R3 is H, alkyl of 1 to 20 ~arbons, arylalkyl, or halogene,
and
X is H or alkyl of l to 20 carbons, C0-S~, CO-O~
CO-NH-~, PO(OS~)2 or PO(OS ,)~ ,, where S ,independently
is ~, alXyl of 1 to 20 carbons, phenyl, or substituted
15 phenyl. However, in the compounds of the invention
illustrated by Formula 2, both ~ and ~3 cannot be
hydrogen.
The present invention also covers salts of the above-
defined compounds, formed with pharmaceutically acceptable
20 acids or bases, as applicable.
In a second aspect the present invention relates to
pharmaceutical formulations comprising one or more
compounds o~ Yor~ula 1 or of Yor~ula 2, or both, (or
pharmaceutically acceptable salts thereof) in ad~ixture -
25 with ~ pharmaceutically acceptable excipient, for the
purpo-~ of tre~ting certain conditions, ~yndro~es or
diseases in ma~mals, including humans. Th~ compound~ of
the invention have anti-infl~mmatory, i~munosuppressant
and anti-prolifQrative activity. ThQrQfor~, the compounds
30 are uBeful for treating in ~ammal8 (including human~)
inflam~ation, rh~u~toid arthriti~, o~teo~rthrlti~,
rheumatic carditis, ocular and dermal infl~mmatory
diseasQs~ autoi~mune diseas~. such a~ allergic di eases,
.. ~ . .. . . .. .
:. . ~
WO91/1605~ 2 ~ ~ 8 7 7 ~ PCT/US9t/02005
bronchial asthma and myasthenia gra~is, and for
suppressing unwanted immune responses and retarding
proliferation of cell.
. ~ R,CHO ~HC_Ca~Et E~ULl ~ R~ OH)C--CCOaEt
Formula 3 4 . Formula 4
1. H2 / Llndlar .
catalyst
2. acld
:
=
o~o~Rl
Formula 1
R2 ~ Ra = H
R-~ctlon Sch~m- 1
In still another a~pect, the present invention
relates to the processes of making the compounds of .
For~ula 1 and of ~o~ula 2. In general tercs, these
processes, shown in R-~ction ~oh-u- 1, 2, 3 ~n~ ~ comprise
the steps of reacting an aldehyde o~ ~o~ula 3 with a
.... ,, . ~ ........ .. . .
, ~ . . . . . . .
WO91/160S5 2 ~ ~ 3 ~ 7 ~ PCT/US91/02005 -
lithium or magnesium salt derived fro~ a propiolate ester, ,
such as ethyl propiolate (Co~poun~ 4), to yield an Rl-
substituted 4-hydroxy --ynoate of For~ula 4 (Rl defined as
in conn~ction with For~ula 1 and Fo~ula 2).
Hydroganation over a "poisoned" catalyst (Lindlar
catalyst) converts th~ acetylenic (triple) bond into an
olephinic (double) bond. This is followed by acid
catalyzed cyclization to yield compounds of For~ula 1
where R2 and ~3 ar~ hydrogen.
~c~ol~c=cc~ KOH , R,C~OH)C_CCO~N
Formub ~ Formub S
. :
Jon-~
r-~-nt
R,COC----CC02H
Formul~ 6
, H2 / Llndbr o~l~ly~t
¦ R~COCH=CHCO,H ¦
¦ cycll2~tlon
R~ccllon Sch-m 2
~,.
D,C;~R,
O 01~ :
Formub 2
R~ ~ R~ H ~ - -
... : : ~ . .. . - . :. : .~ ., , -
W091/16055 2 ~ 7 ~ 7 7 ~ PCT/US91/02~S
In order to obtain compounds of For3ul~ 2, as is
shown in Re~ct~o~ 80heme 2, the intermediate Rl-
substituted 4-hydroxy--ynoate of For~ula ~ is saponified
to obtain the corresponding free carboxylic acid of
~ormula 5, which is thereafter oxidized (typically with
Jones reagent) to yield the corresponding Rl-substituted
4-oxo-alkynoic acid of For~ul~ 6. The intermediate of
Formula 6 is hydrogenated over a "poisoned" catalyst,
typically Lindlar catalyst, to yield an alkenoic acid
intermediate which cyclizes, usually spontanously, to
provide compounds of Fo~mula 2 where R2, R3 and ~ are
hydrogen. As is shown in R-~otion 8ch~m- 2, the compounds
of Formu~a 2 can be reduced with a mild redu~ing agent,
such as sodium borohydride to provide the corresponding
compounds-of ~ormul~ l. A theoretical explanation for
this reaction (although the present inventors do not wish
to be bound by theory) is that the carbon in the 5-
position of the 5-hydroxy-2(SH)-furanone molecule is an
"aldehydic" carbon, ring closed with the carboxylic acid
group in the "2-position" of the ring, and that the
aldehydic carbon is reduced with 80diu~ borhydridQ to a
primary alcohol, which, thereafter., ring closes w~th the
carboxylic acid to for~ a lactone of Yor~ula ~.
Compounds Or ~or~ula 2 where the S substituent is
other than hydrogen, can be obtained by acylation,
carbamate formation (through rQaction with isocyanate)
phosphorylation and the like, in accordance with
procQdures which are within the sXill of the practicing
organic chemist.
.
' - - . . ' . ~
' :: '. ' . , ' ' ' . ' ~ , ' :
, , . -. . , : . . ~
' . : ,- . .. ' : ' ' .: . ' :
WO 91/16055 2 ~3 7 8 7 r~t ~ pcr/us9l/o2oo5 ~
R~COCH2R2 ~CHOCO2H acld ~¦ R~coCR2=cHco2H ¦
Formula 7 ~ Formula 8
S ~ ''
/ heat
~/
10 ~, , XO~( ~, NllrH~ D~, ;
Formula 2 Formula 2 Formula 1
X = alkyl R3 = X = H R3 = H
R3 = H
Br2 . ,
L~R H2O / acid ~ ~ 1
20 o ox O OH
Formula 2 Formula 2
X = alkyl
Reactlon Scheme 3
WO91/16055 2 ~3 7 g ~ 7 ~ PCT/US91/02005
In order to obtain compounds of Formula 2 where
neither R~ nor ~2 are hydrogen, a ketone of Formul~ 7
(which bears the desired Rl and R2 groups) is condensed
under acidic condition with glyoxylic acid (Compoun~ S),
as is shown in ~eAct~o~ 8chQ~- 3. An inter~ediate
condensation product of Formula 8 usually is not isolated
because it cyclizes during the condition of the
condensation reaction to form the furanones of For~ul~ 2
where the Rl and R2 groups are derived from the starting
ketone of For~ula 7 and where S is hydrogen. Reduction of
the 5-hydroxy-2(5H)-furanone compound of For~uls 2 (S is
~) with sodium borohydride, as shown in ~e~ctlon 8ch-~e 3
results in "removal" of the 5-hydroxy group and yields the
corresponding compound of For~ula l.
Referring still to R-actlon 8c~ 3, compounds of
Formul~ 2 where Rl is hydrogen, are obtained when the
starting material of Formul- 7 is an aldehyde rather than
a ketone, (in Formula 7 Rl-H). In this case, the
condensation reaction with glyoxylic acid (Co~pound 5) is
usually performed in the presence of morpholine
hydrochloride. An alkyl substitutent for the 5-hydroxy
function (in Formula 2 ~alkyl) is introduced into the
molecule by reacting the 5-hydroxy-2(5H)-furanone
derivative of For~ul- 2 with the appropriate alcohol (SO~)
in the presence of acid. The resulting 5-alkoxy-2(5H)-
furanone derivative Or Tor~ul~ 2 where R3 i5 hydrogen, is
brominated to yield the corresponding 3-bromo-5-alkoxy-
2(5H)-furanone derivative Or For~ula 2. The alkoxy
substituent can be replaced wit~ OH by reaction with
agueous acid.
.: . . . . . ~ - -
.
2 ~ ~ 8 7 7 ~L
WO 91/160S5 PCI-/US91/02005 _
R~ ~ CH5Cil,1 BuLI ,~c~CR,'
Formula 9 Formula 10 Formula 11
RlMgBr H2
~ ~ ~ ~
R~ R2 R3 R2
O~OH O~OH
ls Formula 2 Formula 2
Na B H4 NaBH4
,,
~ ~
Formula 1 Formula
:
Reaction Scheme 4
W O 91/16055 2 ~ r~ ~ 7 7 ~ PC~r/US91/02005
Re~ct~on 8c~me ~ summarizes a reaction sequence, in
which compounds of the invention can be prepared where
neither R2 nor R3 are hydrogen. In accordance with this
scheme, a di-substituted maleic acid anhydride (For~ul~ 9)
S is reacted with the lithium salt of an alkyne (Forsul~ 10)
to provide a 5-alkynyl substituted 5-hydroxy-2(5H)-
furanone derivative (Formul~ 11). In Formul~ 10 -CC-R~
symbolizes such a precursor of the group Rl which is
readily converted by hydrogenation and or other reactions
within the skill of the practicing organic chemist, into
the group R~ defined in connection with Formula 1 and -
Formul~ 2. Reactlon 8c~-m- ~ show~ a hydrogenation step
in which the olephinic bond of the intermediate of Formul~
11 is partially or fully saturated to provide compounds of
Formul- 2. Reduction of the compounds of Fon~ul~ 2 with
sodium borohydride provides compounds of Formula:l.
Another process of preparing compounds of the
invention where neither R2 nor ~3 are hydrogen, involves
the react~on a di-substituted maleic acid anhydride of
Formula 9 with a Grignard reagent of the formula Rl-MgBr
to provide compounds of ror~ul~ 2 where S is hydrogen.
Reduction of the~e compounds with sodiu~ borohydride,
yields compounds of Yor~ul~ ~.
In anothe~ general aspect, the processes leading to
the compounds of the invention ~ay involve performance of
routin~ chemical reactions (~uch as e~terification,
saponlfication of esters, oxidation of alcohols to ketones
or aldehydes, formation o~ acetals, ketal , and lactones
from aldehydes or keton~s, and th~ like) which are well
known to the practicing synthetlc organic chemist.
When it i5 desired to ~ubstitut~, for example
acylate, the 5-hydroxy function of th~ compound3 of
Formula 2, an S group (as defined in connection with
.: :
.- . . ~ . .
. , .
. . , . ~ .. ,; .. .
2~7877P
W091/16055 PCT/US91/02~5.
these formulas) can be introduced into the
5-hydroxy-2(5H)-furanone compounds by conventional means.
As it was illustrated in connection with R-~ct~o~ schQme
3, introduction of an ~ alkyl group into the compounds of
Formul~ 2 in reality is formation of an acetal, which can
be accomplished by treating the respecting
5-hydroxy-2(SH)-furanone with an alcohol 20H in the
presence of acid.
General Emb~dim~
10 Definitions
The terms "ester", "amine", "amide", "ether"
"acetal", "lactone" and all other terms and terminology
used here, (unless specifically defined in the present
description) refer to and cover any compounds falling
within the respective term as that term is classically
used in organic chemistry.
Unless specifically noted otherwise, preferred esters
are derived fro~ the saturated aliphatic alcohols or acids
of ten or.fewer carbon atoms or from the cyclic or
saturated aliphatic cyclic alcohols and acids of 5 to 10
carbon atoms. Particularly preferred aliphatic esters are
those derived from lower alkyl acids or alcohol~. Also
preferred are the phenyl or lower alkylphenyl esters.
The term ~alkyl" as used in the pre6ent description
and claim~ includes straight chain alkyl.group~, branched
chain alkyl group~, cycloalkyl groups, alkyl .ubstituted
cycloalkyl groups, and cycloalkyl ~ubstituted alkyl
groups. Unles~ the nu~ber of carbons i~ otherwise
specified, ~10WQr alkyl~ means the ~ormer bro~d
definition of "al~yl" groups ~ut with tha re~triction that
~he group has 1 to 6 carbon atoms.
The term "long chain alkyl" ~l~o ~e~n~ the for~er
broAd def~nition o~ "alkyl" groups ~ut with the
. . . . . . .
.
'
WO91/16055 2 ~ 7 o 7 7 ~ PCT/US91/02005
restriction that the group has no less than 4 carbon
atoms, and no more than approximately 2~ carbon atoms.
Unless specifically noted otherwise, preferred amides
are the mono- and di-substituted amides derived from the
saturated aliphatic radicals of ten or fewer carbon atoms,
or the cyclic or saturated aliphatic-cyclic radicals of 5
to lO carbon atoms.
Some of the compounds of the invention (~or~ula 1)
contain a non-equilibrating chiral center in the 5
position of the furan ring. Other compounds of the
invention may contain one or more additional chiral
centers. Accordingly, the compounds of the invention may
be prepared as mixtures of enantiomeric compounds (where
the enantiomers ~ay or ~ay not be present in equal
amounts) or ~s optically pure enantiomers. When there is
more than one chiral center, the compounds of the
invention may also be prepared as mixtures of
diastereomers, or as pure diastereomers, and each
diastereomer itself may be a mixture of enantiomers in
1:1, or other, ratios. Alternatively, each diastereomeric
compound may be sterically and optically pure. However,
all of the abovo-noted form~, including optically pure
enantiomers and mixtur~s thereof, a~ well as all
diasterQomers, are within scope of the pre3ent invention.
Som~ of the compound~ of the invention, for example
tho~e which contain olephinic doubl~ bond~ in the side
chain~, may have Çi~ and tran~ storQoi~om~rfi. The scope
of the invention includes both pur~ ster~oisomers as well
a~ mixtures thereof.
A pharmacQuticalIy acc-pt~bl~ salt may be prepared
for ~ny compound of this invention having a functionality
capable of forming such salt, for ex~mle an acid or an
amine functionality. A pharmac~utically acceptable salt
-
- .. . . : .
. :
2 ~ 7 ~
W O 91/16055 PC~r/US91/02005
may be any salt which retains the activity of the parent
compound and does not impart any deleterious or untoward
effect on the subject to which it is administered and in
the context in which it is administered.
Such a salt may be derived from any organic or
inorganic acid or base. The salt may be a mono or
polyvalent ion. Of particular interest where the acid
function is concerned are the inorganic ions, sodium,
potassium, calcium, and magnesium. Organic amine salts
may be made with amines, particularly ammonium salts such
as mono-, di- and trialkylamines or ethanol amines. Sal~s
may also be formed with caffeine, tromethamine and similar
molecules. Where there is a nitrogen suf~iciently basic
as to be capable of forming acid addition ~alts, such may
be formed with any inorganic or organic acids or
alkylating agsnt such as methyl iodide. Preferred salts
are those for~ed with inorganic acids such as hydrochloric
acid, sulfuric acid or pho~phoric acid. Any of a number
of simple org~nic acid~ such as mono-, di- or tri-acid may
also bQ usQd.
The prefQrred compounds of the present invention are,
with referencQ to ror~ula 2 and with re~p~ct to the OS
substituent in thQ 5-position of the furanone moiety,
tho~e where thQ sub~tituent is hydroxy, ~thoxy or
25 ac~tyloxy (~ i~ H, or CH30 or CH3CO). -
Wlth r sp~ct to the Rl susbtituQnt in th~ 5 position
of th~ 2(5H)-furanone molecule, th~ prQferred compounds in
accordance with ~he pr~sent invention are those where ~1
is hydrogen, long chain alkyl, or ~rylalkyl. Compounds
ar~ particularly pr-ferr~d in thi8 rQg~rd whQre th~ ~1
group i~ long chain alkyl which i~ fitraight chain-d, or
where the ~ i8 ~rylalkyl containing a straight ~lXyl
chain of 3 carbons.
-: ., , : . ... ,-.. , ... .. ,. .... , ~.. - .. ,~: - ... .. ~ . -
WO91/16055 2~7~77 1 PCT/US91/02005
With respect to the ~2 substituent in the 4-position
of the 2(~H)-furanone molecule, the compounds of the
invention are preferred where ~2 is hydrogen, or alkyl
group, particularly straight chain alkyl
With respect to position 3 of the 2(5~)-furanones of
the invention, compounds are preferred where ~3 is H,
methyl or bromo
The most preferred compounds of the invention are
those listed just below with reference to Formul~ 1 or
10 Formul~ 2
Formula 1, compoun~ 6 ~1=C~3(CH2)8~ R2-H~ and R3 H;
Formula 2 Co~poun~ 7 R1=CH3(CH2)g~ R2 H, R3 H
S-H
Formul~ 1, compoun~ 8 ~1=(CH2)3-c6H5~R2=cH3~ and
15 R3-C~3;
' P ~ 9 R1 H~ R2-CH3(CH2)7~ R3=Br and
SSH, and
~or~ul~ ', Co~poun~ lO Rl'H~ R23C~3(CH2)7 and R3 H
The compounds of the present inv~ntion are useful in
pharmaceutical compositions to produce anti-inflammatory,
immunosuppres~ant and anti-proliferative activity The
diseases, syndromes or conditions of mammals (including
humans) which can be treated with pharmaceutical
compositions containing one or more co~pounds of the
invention (or salt~ thereof) include inflammation,
rheumatoid arthriti~, osteoarthritis, rheumatic carditis,
ocular and dermal $nflammatory diseasQs, autoimmune
di~ease~ 3uch as allergic disaases, bronchial asth~a and
myasthenia gravi~, unwanted immune r~pon~es and unwanted
proli~eration of cell~, psoria~i~, acne, atopic diseases
and allorgic con~unctiviti~
The activ~ty o~ the compounds o~ this invention is
demonstrated by ~nhibition of the enzyme phospholipase A2
.. . . . . . . ....................... .
.. .. . .
. - .. - . .. .. . ., ~, . . ~ . . ~
WOsl/16055 2 ~ ~ ~ 7 7 ~ PCT/US91/02005 ~
.~ vitro and by reduction of inflammation in the mouse ear
anti-inflammatory assay in vivo.
Activity of compounds of this invention may also be
demonstrated by inhibition of phosphoinositide-specific
phospholipase C. This activity has been reported for
manoalide and may indicate anti-inflammatory utility.
Bennett et al, Molecular PharmacolQgy 32:587-593 (1987).
Activity of the compounds may also be demonstrated by
inhibition of ornithine decarboxylase, a rate limiting
enzyme in cellular growth, which indicates use in treating
psoriasis and neoplasis.
The compounds also modify calcium homeostasis. This
activity i8 shown by effect on intracellular calcium
levels in experiments using gastric glands, spleen cells,
epithelial cells, GH3 cells, etc. Calcium is inhibited
from entering through the plasma membrane calcium channels
and calcium release from intracellular stores is also
blocked. Modification of calcium homeostasis is expected
to have application in diseases of the nervous ~ystem
involving modification of membrane lipids or transmitter
release (Parkinson's, Alzheimer's), di~eas~s of the
cardiovascular sy~tem involving application of cardiac or
vascular smooth muscle contractility and platelet
aggregation (hypertension, cardiac infarction and
atherosclerosi~), diseases of the ga~troint~stinal tract
such a8 ul¢er disease, diarrhea, motility due to secretion
of acid or Cl , diseases of the kidney involving renal
handling of fluid an~ electrolytes tmetabolic acidosi~,
alk~lo~is), and di~eas~ of abnormal growth (neopla~ia,
psoria~is).
The compounds of thiQ invention hav~ acti~ity which
i~ si~ilar to that o~ manoalide, th~t i8 the co~pounds
app~ar to be devoid of the endo~rine propertie~ of the
. , ~ , . , -
. . . . .. ..
... . ... , . ~, , , : , ..
- . . .. . . . , :
.: .: ~ . .
.. . . . .
- . ~ ..... ,. . ~, ;: , - . . ~
WO91/16055 PCT/US9t/02~5
2~7~7 ~
17
glucocorticoids while having anti-inflammatory and
immunosuppressive properties.
In the method~ of this invention, the compounds of
the invention are administered to mammals, includi~g
humans, in an effective amount to produce the desired
activity, preferably in an amount of about 0.05 to lOO mg
per day per kilogram of body weight. The amount of the
compound depends upon the disease or condition being
treated, the severity thereof, the route of ad~inistration
lG and the nature of the host. The compounds may be
administered topically, orally, parenterally or by other
standard routes of administration.
Pha~maceutical compositions of this invention
comprise compounds of Yormula 1 and of For~ul~ 2, and
pharmaceutical carriers suitable for the route of
ad~inistration. Standard methods for formulating
pharmaceutical compositions o~ this type may be found in
Reminaton's Pha~se~dh~LL~ ~ ss~ Mack Publishing
Company, Easton, PA.
For topical administration, the pharmaceutical
composition may be in the for~ of ~ salve, crea~,
ointment, spray, powder or the like. Standard
pharmaceutical carriers for such compositions may be used.
Preferably, compo~itionQ for topical administration will
contain 0.05-5% of the active ingredient.
A typic~l cream formulation may contain the
following: -
Inaredient - P~rts by Weiaht
-Water/glycol mixture 50-99
(15% or ~ore glycol)
Fatty ~lcohol 1-20
Non-ionid surfactant 0-lO
M~neral oil 0-lO ;
, ~
. . ...... . . , . . : . . , . ,., -
W091/160S5 2 ~ 7 8 7 7 L PCT/US9t/0200s.~
18
Typical pharmaceutical adjuvants 0-5
Active ingredient 0.05-5
A typical ointment formulation ~ay contain the
following:
Inaredients ~ s by Weiht
White petrolatum 40-94
Mineral oil 5-20
Glycol solvent 1-15
Surfactant 0-10
Stabilizer O-lo
Active ingredient 0.05-5
For oral ad~inistration, suitable pharmaceutical
carriexs include mannitol, lacto~e, ~tarch, ~agnesiu~
stearate, talcum, glucose and magne ium carbonate. Oral
compositions ~ay be in the form of tablets, capsules,
powd~rs, solutions, suspensions, su6tained release
formulations, and the like.
A typical tablet or capsule may contain the
following:
Inaredients P~çent w~w
Lactose, ~pray-dri~d 40-99
Magnesium ste~rat~ 1-2
Cornstarch 10-20
Active ingredient 0.001-20
Parenteral compo~ition~ are prepared in conventional
suspen~ion or solution ~orms, a~ emuls~ons or as solid ~ :
forms ~or reconstruction. Suitable carriers are water,
saline, dextro--, Hank' 8 solution, ~inger'~ solution,
glyc-rol! and the like. Parent~ral administration i~
usually by in~ction which ~ay be ~ubcutaneous,
intramuscular or intravenous.
., . .. . ;
WO91/16055 2 ~ ~ ~ 7 7~ PCT/US91/02005
The compounds of this invention may be combined with
other known anti-infla~matory/immunosuppressive agents
such as steroids or non-steroidal anti-inflammatory agents
(NSAID) in the pharmaceutical compositions and methods
deecribed herein.
The assay procedures by which useful biological
activity of the compounds of the invention can be
demonstrated, are described below.
Calcium Channel (mobilization~ Inhibition Assay
Polymorphonuclear leukocytes (PMNa), gastric glands,
GH3 cells, A431 cells, spleen cells, hu~an ~eratinocytes
corneal cells, etc. were loaded with the Ca2+ sensitive
fluorescent dye, Fura-2. The appropriate cell type was
chosen and the potency and e~ficacy of the anti-
inflammatory furanones on calcium mobilization, calcium
channel inhibition wa~ quantitated. The methods used for
A431 cells listed below are representative of those used
for other cells.
A431 cell~ were detached using a 5-10 min trypsin-
EDTA trea~ment wherQas GH3 cell~ were treated 2 to 5 minwith a 1% pancreatin solution. Cells were immediately
washed twice in a 20mM HEPES bu~fer (pH 7.4) containing
120mM NaCl, 6 mM KCl, 1 ~M MgS04, 1 mg/ml gluco~e and
mg/ml pyruvatQ and 1.4mM calcium (mediu~ A).
Approximately 5 x 106 cell~ w~re suspQnded in medium A and
incubated with 4uM fura-2-AM for 15 ~in ~t 37C.
After washing the ~ura-2 loaded cells, the uptake of
dye was checked using fluorescence microscopy and found to
be evenly ~istributed in the cyto801 0~ all cells.
Fluore~cenc~ waB continucusly r~cordsd with a Perkin-Elmer
LS-5 spQctrofluorom~ter. The excltation w~velength was
set at 340n~ and emicsion wavelength ~et at 500nm. The
cell suspension was continually stirred, maintained at
~ . - - .. .. . . . . . . ......... . . . . ................ .
.. .. : . - - . . , . ~ . . . : . . ~ .
W091/16055 2 ~ 7 ~ 7 7 ~ PCT/US9t/02~5 ~
37C and equilibrated for approximately 5 min before
addition of various agents. [ca2+i was calculated using
the following formula:
tca2+]i = 220 X F - Fmin
Fmax - F
All fluorescence values were measured relative to a
EGTA-quenched signal determined as follows: F was the
relative fluorescence measurement of the ~ample. FmaX was
determined by lysing the cells with digitonin (lOOug/ml)
in DMSO. After FmaX was determined the pH was adjusted to
8, with NaOH and Ca2+ chQlated with 3mM EGTA to totally
quench the fura-2 sis ~1 and obtain Fmin.
When quin-2- was used, cell~ were incubated with lOuM
quin-2- at 37C for 1 hour, wa~hed and th~n used.
is Mouse ~ an~i-Inflam~tory Assay
Test compound and phorbol ~yrist~te ~cetate (PMA) are
topically applied simultaneously to the pinnae o~ the left
ears of mice. PMA alone is applied to the right ear.
Three hours and 20 minute~ after application, the mice are
sacrificed, left and ri~ht e~r~ remo~ed, and standard
sized bores taken. Edema (inflammation) is m~asured as
the diffQrence ~n weight between left and right ears [Van
Arman, C.G., Clin Phar~col ~h~ (1974) 16:900-904].
Inhibition o~ PhoQpholipase A2
Th efSect of compounds of thi~ invention on bee
vonom phospholipa~ A2 i d~termin~d by the following
procedure:
a. Bee venom pho~pholipasQ A2 in lO uM HEPES (pH
7.4)
with 1 mM CaC12 i~ incubat~d with v~hicl~ or
test ~gent for l.O hour ~t 41.
b. 1.36 mM pho~photidylcholine, 2.76 mM Triton X-
:. .. : . ~ -
- ~ , .. :, ~.
WO 91/16055 2 ~ ~ ~ 7 7 ~ PCT/US91/02005
loo ?
are dispersed in buffer by sonication and then
mixed with L-3 phosphotidylcholine, l-palmitoyl-
2-
s (1-14C) palmitoyl for 10 min.
c. Start the reaction by the addition sf enzyme
(0.495 units/ml).
d. Incubation for 15 sec. at 41.
e. Reaction is terminated by addition of 2.5 ~1 of
isopropanol: n-heptane: 0.5 M H2SO4 (40:10:1;
v:v:v: ) .
f. 2.0 ml n-heptane and 1.0 ml H2O added; mixture
centrifuged.
g. 2.0 ml n-heptane removed and treated with 200-
300 mg
of silica gel HR60.
h. Samples centrifuged; 1 ml of n-heptane SN
removed and
added to 10 ml scintillation fluid.
i. Samples counted on a scintillation counter.
Inhikition.of Phos _ cific Phospholi~ase C
The ef~ect of compound~ o~ this in~ention on
phosphoino~itide-~pecific phospholipase C ~ay be
determined by procedures described by Bennett et al,
Molecular Pharmacology 32:587-593 (1987).
Activ~ty Data
In the above-de cribed pho~pholipase A2 assay the
-- compounds of the invention we~e found to provide 50%
inhibition (IC50) o~ bee vsnom phospholipa~e A2 at the
following concentrations (in micromol0~ indicat~d in
~1- 1. ' '
~'..
wO91/16055 2 0 7 ~ 7 ~ ~ PCT/US91/02~5 ~
T~ble 1
Phospholipase A2 Assay.
Compound name or number IC50 (um)
l 0.03
7 6
*Data for Compoun~ l (monoalide) are provided for
comparison.
In the above-described phospholipase A2 assay
compounds of the invention which are listed in ~abl- 2
were found to provide the following percentage (%)
inhibition of bee venom phospholipase A2 at lO0 micromolar
concentration.
~4b~0_2
.
6 37
8 49
53
The compounds of the present invention can be made by
the synthetic chemical pathways which are illustrated
above in general terms, and in the ~pecific examples as
well. The ~ynthstic che~ist will readily appreciate that
the conditions described here in general terms, and
specifically, can be generalized to any and all co~pounds
repr-sented by ~or~ul~ ~ or by For~ul~ 2, as applicable.
Furthermore, th~ synthetic ch~mist will readily appreciate
that the hereln dQscribed ~ynt~etic step~ may be varied or
adjusted by tho~e ~killed in the ~rt wlthout departing
from the soope and spirit o~ the invQntion. Therefore,
the following examples o~ spec~ic compounds of the
invention, and spec$fic examples of the synthetic steps in
which the compound~ and cert~in intermediates are made,
, ~ ~}
- . , : . .
-, . . . . . . .. . .
.. : ...... :; : : , . .. . . . : . . . .
WO91/16055 2 ~ 7 ~ 7 71 PCT/US91/02005
are set out to illustrate the invention, not to limit its
scope.
Specifi~ E~mp~es
Example 1
Ethyl 4-hydroxy-6-phenylhex-1-ynoate (Compoun~ 11)
n-Butyl lithium (a 1.6 M solution in hexane; 6.7 ml,
10.7 mmol) was added dropwise to a solution of ethyl
propiolate (1.04 g, 10. 6 mmol ) in tetrahydrofuran (10 ml)
at -78 under argon. After 10 minutes, a solution of
hydrocinnamaldehyde (1.42 g, 10.6 mmol) in tetrahydrofuran
(5 ml) was added. Stirring was continued at -78C for 2
hours and acetic acid (1 ml) was added. on warming up to
0, the reaction mixture was poured into water.
Extraction (ethyl ether) and evaporation of the dried
(magnesium sulphate) extracts a~forded an oil, which was
flash chromatographed with silica using 30% ethyl
ether/petroleum ether. Fractions with Rf of about 0.28 on
evaporation afforded the title ~ster as a light yellow
oil.
1 NMR (CDC13) 1.34 (m, 3H), 2.10 (m, 2H), 2.40 (br,
lH), 2.83 (t, 2H, J - 8.3 Hz), 4.25 (q, 2H, J s 7.3 Hz),
4.50 (t, lH, J ~ 7.0 Hz) and 7.29 (m, 5H).
LRNS m/e (% abundance) 233 (M++1, 2), 232 (M+, 7),
186 (24), 185 (42), 170 (51), 169 (30), 158 (24), 142
(37), 141 (100) and 105 (84).
5-~2-Ph~ny~a~hy~-2f5H)-furanone (Compou~ 12)
A ~olution of ethyl 4-hydroxy-6-phenylhex-1-ynoate
(Compo~d 11, 585 mg, 2.5 mmol) in ether (12 ml) was
hydrogen~ted over ~indlar catalyst (50 ~g) at roo~
temperature for 3 hour~. The ~ixturQ wa8 ~ltered through
celite and the filtrate was refluxed w~th 2M hydrochloric
acid (1 ml) for 2-1/2 hours. On cooling, the mixture was
. ~ . . . , . . . . . . . . , .~ - , .
. , .. . ,. ,- . . . -, .
. . , .. , . , : .
Wosl/l6o55 2 ~ ~ ~ 7 7 ~ PCT/US9l~02005-~ 1
24
dried (magnesium sulphate) and evaporated to dryness to
give an oil, which was purified by preparative thin layer
chromatography (tlc, 20x20 cm, 2000u silica plate:
developed with 30% ethyl ether/petroleum ether). The
title furanone was obtained as colorless prisms
(recrystallized from ether): mp 66-7.
lH NMR (CDC13) 1.98-2.16 (m, 2H), 2.86 (m, 2H), 5.08
(m, lH), 6.16 (dd, lH, J = 6.0 Hz, 1.6 Hz), 7.36 (m, 5H)
and 7.45 (dd, lH, J s 6.0 Hz, 1.6 Hz).
13C NMR (CDC13) 31.3, 34.9, 82.3, 121.5, 126.3,
128.5, 128.6, 140.2, 156.1 and 172.9.
HRMS exact mass calculated for C12H1202 (M+)
188.0837, found 188.0841.
Exam~le_~
4-Hy~oxy-6-phenylhex-2-ynoic acid (Co~pou~ 13)
A solution of potassium hydroxide t377 mg, 6.7 mmol)
in 95% ethanol (lo ml) was added to a solution of ethyl 4-
hydroxy-6-phenylhex-1-ynoate (Co~pou~ 11, 1.04 g, 4.5
mmol) in the ~ame solvent tlO ~1) at 0, and the reaction
mixture was stirred at room temperature for 15 hours.
After most o~ the sol~ent was removed, the residue was
dissolved in water (ca. 15 ml) and ex*racted with
dichloromethane (discarded). After the extraction the
aqueous phas~ w~ acid~fied to pH 1 with dilute
hydrochloric acid ~nd extra~ted thorou~hly with ethyl
acetate. Ev~poration of the dried (magnesium sulphate)
extracts gave the titl~ ac~d ~8 a pale yellow oil (which
cryst~llizes 810wly on standing), which w~s used directly
in the next step.
lH NMR (CDC13) 2.16 (m, 2H), 2.85 (m, 2H), 4.52 (dd,
lH, J - 11.3 Hz, 6.6 Hz), 5.10 tbr, 2H) ~nd 7.31 (~, 5H).
LRMS ~/e (% ~bundance) 204 (M+, 6), 142 (42), 141
(75), 134 (21), 133 (11), 131, (10), 118 (34), 11~ (32),
I
Wo9l/l6055 2~o771 Pcr/lls9l/n2l)os
115 (21) and 105 (100).
4-Keto-6-phenylhex-2-ynoic acid (Co~pou~ )
A solution of Jones Reagent (a 2.67 M solution in
sulphuric: acid; 2.07 ml, 5.5 mmol) was added dropwise to a
s solution of 4-hydroxy-6-phenylhex-2-ynoic acid (~ompound
13, 750 mg, 3.7 mmol) in acetone (12 ml) at 0 and the
reaction mixture was maintained at 0 for 70 minutes. The
mixture was quenched with ethanol (2 ml) and extracted
with ethyl ether. Evaporation of the dried (magnesium
10 sulphate) extracts gave the title acid as a yellow oil
which was used directly in the next step.
1 NMR (CDC13) 3.02 (s, 4H), 7.30 (~n, 5H) and 8.80
(br, lH, exchanged with D20).
5-Hydroxy-5-(2-Dhenylethyl)-2-furanone (Compou~ 15)
A solution of 4-keto-6-phenylhex-2-ynoic acid
(Co~pou~ , 228 mg, 1.1 mmol) in ethyl ether t8 ml) was
hydrogena~ed over Lindlar catalyst (20 mg) at 0 ~or 80
minutes. The mixture was filt~red through celite and the
filtrate, after evaporat~on to dryness, was purified by
20 preparative tlc (20x20 cm, lOOOu silica plate; developed
with 60% ethyl ether/hexane). The title furanone was
obtained a~ a colorless oil.
lH N~ (CDC13) 2.31 (dd, 2H, J -- 10.8 H2, 5.5 Hz),
2.78 (dd, 2H, J - 10.8 Hz, 5.5 Hz), 4.80 (br, lH), 6.11
(d, lH, J - 5.8 Hz), 7.20 (m, 5H) and 7.28 (d, lH, J s 5.8
Hz).
13C N~ tC~C13) 29.7, 39.1, 107.8, 123.1, 126.4,
128.3, 128.7, 140.3, 154.3 and 171Ø
HRMS exact mass calculated for C12H1203 (H+) ,, ;
204.07`86, found 204.0792.
EXZ~ 3
2~hY1 4-hydrox~lftridec-2 vno~te (Co~pou~ld ~6)
Methylmagnesi~L~ bromide (a 3M ~olution in
WO91/160~5 2 ~ ~ 3 ~ 7 ~ PCT/US91/02005 _
26
tetrahydrofuran; 7.8 ml, 23.4 mmol) was added dropwise to
a solution of ethyl propiolate (Compoun~ ~, 2.25 g, 22.9
mmol) in tetrahydrofuran (10 ml) at -78 under argon.
After 10 minutes, a solution of decyl aldehyd~ (3.58 g,
22.9 mmol) in tetrahydrofuran (2 ml) was added. Stirring
was continued for 1 hour while the cooling bath was warmed
to room temperature. The mixture was quenched with
saturated ammonium chloride solution and extracted with
ethyl ether. Evaporation of the dried (magnesium
sulphate) extracts gave an oil, which was flash
chromatographed on silica using 30% ethyl ether/petroleum
ether. Fractions with Rf of about 0.31 qave, after
evaporation, the title ester as a deep, yellow oil.
lH NMR (CDC13) 0.88 (t, 3~, J ~ 6.4 Hz), 1.27 (br s,
14H), 1.75 (m, 2H) and 4.25 (q, 2H, J - 6.4 Hz).
LRMS m/e % abundance) 255 (M++l, 5), 254 (M+, 5), 237
(6), 209 (8), 181 (12~, 179 (11), 163 (13), 152 (12), 151
(13), 137 (16), 130 (19), 128 (100), 100 (66) and 71 (35).
5-~Q~yl-2~5H)-fu~none (Co~poun~ 6)
A solution of ethyl 4-hydroxytridec-2-ynoate
(Compou~ 16, 230.6 ml, 1.02 mmol) in ether (10 ~1) was
hydrogenated o~er Lindlar catalyst (20 mg) at 0 for 1
hour. The mixture was filtered through celite and after
evaporation the filtratQ gav- a residue, which was flash
2S chromatographed on silica u~ing 60% athyl ether/petroleum
ether. Fractions with ~f of about 0.18 gave after
evaporation a colorles~ oil (157 mg, 59%) identified by lH
NMR as Qthyl (~)-4-hydroxytridec-2-enoate: lH NMR (CDC13)
0.92 (t, 3H, J - 6.7 Hz), 1.31 (br ~, 14H), 1.55 (m, 2H),
4.97 (q, lH, J - 6.2 Hz), 5.91 (d~ 1~, J - 12.5 HZ) nnd
6.41 (dd, lH, J - 12.5 Hz, 7.3 HZ). Th~ oil on
crystallization fro~ petroleu~ ~ther, in the presence of a
drop of acetic acid, lactonized to give the title furanone
.
:' . . . -
.- . . ~ , ~
WO91/1605~ ~ 3 7 ~ 7 7 ~ PCT/US9l/02005
27
as a colorless oil.
lH NMR (CDC13) 0.92 (t~ 3H~ J = 6.7 Hz), 1.30 (br s, r
12H), 1.45 (m, 2H), 1.75 (m, 2H), 5.08 (m, lH~, 6.14 (dd,
lH~ J = 5.9 Hz~ 2.6 Hz) and 7.48 (dd, 1~, J = 5.3 HZ~ 1.4
5 Hz).
13C NMR (CDC13) 14.1, 22.6, 25.0, 29.2, 29.3, 29.4,
31.8, 33.2~ 83.4, 121.5, 156.3 and 173.1.
HRMS m/e: exact mass calculated for C13H2202 (M+)
210.1620, found 210.1624.
10 ExamplÇ 4
4-Hydr~ytridec-~-ynoic acid (compoun~ 17)
A solution of potassium hydroxide (885 mg, 15.8 mmol)
in 95% ethanol (35 ml) was added to ethyl 4-hydroxytridec-
2-ynoate (Co~poun~ 16, 2.68 g, 10.5 m~ol) in the same
15 solvent (5 ml) at 0. After ~tirring at room te~perature
for 9 hours, most of the solvent was removed and water (20
ml) was added. ThQ mixture was extracted thorouqhy with
dichloromethane (discarded), acidified to pH 1 with dilute
hydrochloric acid and extracted with ethyl acetate.
20 Evaporation of the dr~ed (magnesium sulphat~) ethyl
acetate extracts gave an off-whitQ solid, which on
recrystallizat~on from petroleum ether (at -78) ga~e the
title acid ~8 colorless p~isms: mp 65-6.
H NMR (CDC13) 0.93 (t, 3H, J ~ 5.7 Hz), 1.31 (br s,
12H), 1.50 (br, lH), 1.81 (~, 2H), 4.56 (dt, lH, J ~ 5.0
Hz, 1.9 ~z) and 5.00 (br, lH).
L~MS m/s ~% ~bundance) 226 IM+, 5), 137 (13), 124
(12), 121 (18), 107 (26), 100 (94), 97 (33), 95 (27), 93
(43) 85 (48), ~3 (63), 79 (55) ~nd 71 (75).
4-K~Q~id~c-2-ynQ~c-ac~ (Compou~ 18) r
Jon~ re~gQnt (a 2.67 M ~olution ~n ~ulphuric acid;
1.32 ml, 3.5 mmol) was ~dded dropwi~e to a solution of 4-
hydroxytridec-2-ynoic acid (~o~pou~ 17, 531.8 ~g, 2.4
2~i37~ ~
WO91/16055 PCT/US91/02005 -
28
mmol) in acetone (lo ml) at o and the reaction mixture
was maintained at 0 for 70 minutes. The mixture was
quenched with ethanol (1 ml) and dried with magnesium
sulphate. on evaporation, the title acid was obtained as
a colorless oil.
lH NMR (CDC13) 0.87 (t, 3H, J = 6.6 Hz), 1.25 (br s,
12H), 1.67 (m, 2H), 2.64 (t, 2H, J = 7.2 Hz) and 4.90 (br,
lH).
LRMS m/e (% abundance) 224 (M+, 5), 223 (12), 197
(36), 155 (37), 149 (14), 37 (21), 123 tl5), 111 (14), 109
(12) and 97 (34).
~-Hydroxy-5-nonyl-2-furanone (Compoun~ 7)
A solution of 4-ketotridec-2-ynoic acid (Compound 18,
220 mg, 0.98 mmol) in ether (10 ml) was hydrogenated over
Lindlar catalyst (10 mg) at 0 for 80 minutes. The
mixture was filtered through celite and after evaporation
to dryness the ~iltr~te gave an oil, which was purified by
preparative tlc (20x20 cm, lOOOu silica plate; developed
with 60% ethyl ether~petroleum ether). The title furanone
was obtained as colorless pri~ms (recrystallized from
petroleum ether): ~p 54-5.
lH NMR (CDC13) 0.88 (t, 3H, J ~ 7.5 Hz), 1.26 (br s,
12H), 1.40 (m, 2H), 1.98 (m, 2H), 6.12 (d, lH, J = 6.4 Hz)
and 7.16 (d, lH, J ~ 6.4 Hz).
13C NMR (CDC13) 14.1, 22.6, 23.4, 29.3, 29.4, 31.8,
37.5, 108.5, 123.0, 154.5 and 170.8.
HRMS m/e: exact mass calculated 20r C13H2203 (M+)
226.1569, found 226.1568.
Ex~m~J~-5
5-Hydroxy-5-met~yL-4-octyl-2-furanQne (Co~pou~
A mixture o~ 2-undecanone (lO g, 58.7 ~ol),
glyoxylic acid monohydrate (Co~pou~ 5, 5.15 g, 56 m~ol)
and 85% phosphoric acid (10 ml) was war~ed at 80 for 18
- , , , -.
-;; : , ,. . ~ .~ - ,
2 ~ 7 ~
wogl/l60ss PCT/US91/02005
hours. On cooling to room temperature, the mixture was
diluted with ethyl ether/dichloro~ethane (50 ml each) and
washed thoroughly with brine. Evaporation of the dried
lmagnesium sulphate) organic phase gave a yellow oil which
on crystallization from ethyl ether/petroleum ether gave
4-ketotridec-2-enoic acid as colorless prisms.
lH NMR (CDCl3~ 0.96 (t, 3H, J = 7.4 Hz), 1.34 (br s,
12H), 1.72 (p, 2H, J = 7.1 Hz), 2.73 (t, 2~, J = 7.1 Hz),
6.74 (d, lH, J = 15.7 Hz) and 7.22 (d, lH, J = 15.7 Hz).
The mother liguor from the above recrystallization
was concentrated down and was flash chromatographed on
silica using 40% ethyl acetate/petroleum ether. Fractions
with Rf of about 0.1 gave, after evaporation, the title
furanone as a pale yellow oil.
a5 lH NMR (CDC13) 0.92 (t, 3H, J = 7.4 Hz), 1.30 (br s,
12H), 2.45 (s, 3H), 2.79 (t, 2H, J = 7.5 Hz) and 6.56 (s,
lH).
13C NMR (CDCl3) 14.1, 22.7, 26.7, 27.0, 29.2, 29.3,
29.8, 31.9, 124.5, 158.0, 171.3 and 200Ø
HRMS m/e: exact mass calculated for C13H2203 (M+)
226.1569, found 226.1559.
Exam~le 6
5-Methyl-4-octvl-2~5H~-furanone (Compou~d 20)
Sodium borohydride (214 mg, 5.7 ~mol) was added to a
solution of 5-hydroxy-5-~ethyl-4-octyl-2-furanone
(Compoun~ 19, 640 mg, 2.8 mmol) in tetrahydrofuran (15
ml). After ~tirring at room temperature for 80 minutes,
moæt of the'solvent was removed and water (lO ml) waæ
added. Extraction (dichloromethane) and Qvaporation of
the drisd'(m2gnesium sulphate) sxtract~ gave a residue,
which was flash chro~atographed on sil~ca u ing 60% ethyl
ether/petroleum Qther. Fract~on~ with R~ of about 0.23
were evaporated to yield the title furanone as a colorless
... . .
WO 91/l~OS~ 2 ~ ~ 3 7 7 ~ PCT/US91/02005 _
oil, which crystallized slowly on storage at -70.
lH NMR (CDC13) 0.85 (t, 3N, J = 5.4 Hz), 1.26 (br s,
loH), 1.33 (d, 3H, J = 6.9 Hz), 1.47 (m, 2H), 2.26 (m,
lH), 2.83 (m, lH0, 4.33 (q, lH, J = 6.2 Hz) and 5.99 (s,
lH).
13C MMR (CDC13) 14.1, 22.4, 22.6, 29.2, 29.3, 29.5,
29.8, 31.9, 71.1, 113.0, 169.2 and 171.8.
HRMS m/e exact mass calculated for C13H2202 (M+)
210.1620, found 210.1617.
Exam~le 7
(Comoun~ 21)
A mixture of 2-pentanone (17.1 g, 198 mmol),
glyoxylic acid monohydrate (Co~pou~ 5, 8.05 g 88 mmol)
and about 85% phosphoric acid (12 ml) was warmed at ca.
80 for 19 hour~. on coolin~ to room temperature the
mixture was diluted with ethyl ether/dichloromethane (loO
ml, 1:1) and washed thoroughly wit~ brine. Evaporation of
the dried (magnesium sulphate) organic phase gave a yellow
viscous oil, which on crystallization from ethyl
ether/petroleum ether gave 4-keto-hept-2-enoic acid as
colorles~ prisms: mp 100-2.
lH NMR (CDC13) 1.01 (t, 3H, J - 7.9 Hz), 1.73 (p, 2H,
J s 7.1. Hz), 2.70 (t, 2H, J - 7.3 Hz), 6.73 (lH, d, J =
15.8 Hz) and 7.19 (d, lH, J 3 15.8 Hz).
HRMS m/e: exact mass calculated for C7H1003 (M+)
142.0630, found 142.0622.
The mother liquor from the above crystallization was
evaporated to dryness and extracted thoroughly with
petrol~um ether. The combined extract3 were concentrated
and cooled to -20 to give the t~tle ~uranone as colorless
prisms: mp 37-8.
lH NMR tC~C13) 1.09 (t, 3H, J ~ 7.8 Hz), 2.46 (s,
; - . : - -
WO91/160~5 2 ~ ~ 3 7 7 ~ PCT/US91/02005
3H), 2.83 (q, 2~), J = 7.8 Hz) and 6.S7 (s, lH).
13C NMR (CDC13) 13.6, 20.4, 26.6, 124.6, 158.7, 171.2
and 199.9.
HRMS m/e: exact mass calculated for C7H1003(M+)
142.0630, found 142.0622.
Example 8
4-~h~1-5-methYl-2f5H~-furanone (Co~poun~ 22)
Sodium borohydride (646 mg, 17 mmol) was added to a
solution of 4-ethyl-5-hydroxy-5-methyl-2-furanone
(compoun~ 21, 1.21 g, 8.5 ~mol) in tetrahydrofuran (10 ml)
at room temperature. After 1/2 hour, most of the solvent
was removed and water (10 ml) was added. Extraction
(ethyl acetate) and evaporation of the dried (magnesium
sulphate) extracts gave an oil, which was flash
chromatographed on silica using 60S ethyl ether/petroleum
ether. Fractions with Rf of about 0.23 gave after
evaporation, a pale yellow oil, which slowly crystallized
on storage at -20. Recrystallization from ethyl
ether/petroleum ether afforded the title furanone as
colorless prisms: mp 86-7.
1H NMR (CDC13) 1.16 (t, 3~, J - 7.2 Hz), 1.39 (d, 3H,
J z 5.4 Hz), 2.36 (m, lH), 2.84 (m, lH), 4.41 (q, 2H, J =
7.2 Hz) and 6.04 (s, lH).
13C NMR ~CDC13) 13.8, 22.3, 22.8, 71.0, 112~9~ 17~3
and 171.7.
HRMS m/e: exact mass calculated for C7H10O2(M+)
126.0681, found 126.0683.
Exam~Ç_ r9~
3.4-Dimethyl-5-hydroxy-5-~1-octvnyl)-2-furanone (Compou~d
23)
n-Butyl lithium (a 1.6 M solution in hexane; 6.78 ml,
10.9 mmol) was added dropwise to a solution of l-octyne
(1.13 g, 10 mmol) in tetrahydrofuran (7 ml) at 78 under
, ,. ~, ~ , .. ..
.
~' . ~ , '
. .
Wosl/16055 2 ~ ~ 3 7 ~ ~ PCT/US91/0~005_
argon. After 20 minutes, the solution was cannulated
dropwise, under argon, to a solution of 2,3-dimethylmaleic
anhydride (1.30 g, 10.3, mmol) in tetrahydrofuran (15 ml)
cooled at -78. Stirring was continued for 2 hours while
the cooling bath attained room temperature. The mixture
was quenched with dilute hydrochloric acid, diluted with
water (10 ~1) and extracted with ethyl acetate.
Evaporation of the dried (magnesium sulphate) extracts
gave an oil, which was flash chromatographed on silica
using 30% ethyl ether/petroleum ether. Fractions with Rf
of about 0.18 on evaporation afforded a light yellow
viscous oil, which crystallized out slowly on storage at
-20. Recrystallization from petroleum ether gave the
title furanone as colorless prisms: mp 55-6C.
lH NMR (CDC13) 0.85 (t, 3H, J - 7.4 Hz), ~.24 (m,
6H), 1.49 (p, 2H, J = 7.9 Hz), 1.79 (s, 3H), 2.00 (s, 3H),
2.21 (t, 2H, J s 7.2 Hz) and 3.93 (br, lH).
13C NMR (CDC13) 8.4, 10.5, 13.9, 18.5, 22.4, 27.9,
28.4, 31.1, 74.5, 88.2, 98.0, 124.3, 156.9 and 172.1.
HRMS m/e: exact mass calculated for C14H2003 (M+)
237.1491, found 237.1498.
ExamDle 10
3.4-Dimethvl-5-hydroxy-5-(2-Dhenylp~opyl~-2-furanone
(Co~poun~ 2~)
A mixture o~ 3-phenyl-1-bromopropane (521 mg, 2.6
mmol) and magn~aium turnings (66 mg, 2.8 m~ol) in
tetrahydrofuran (5 ml) was refluxed under argon for 90
minutes. After the reaction mixture had been cooled to
-78, a ~olution o~ 2,3-dimethylmaleic anhydride (330 mg,
2.6 mmol) in ~etrahydro~uran (5 ml) wa3 added dropwise.
Stirring wa~ continu~d ovornight (ca. 17 hour8) while the
cooling bath attained room temperature. The mixture was
quenched with a saturated solution o~ am~oniu~ chloride
, .. . . . .. .
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. : .
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WO91/16055 2 8 ~ o ~ 7 ~ PCT/US91/02005
and extracted with ethyl acetate. Evaporation of the
dried (magnesium sulphate) extracts gave an oil, which was
flash chromatographed on silica using 40~ ethyl
acetate/petroleum ether. Fractions with Rf of about 0.32
on evaporation afforded the title furanone as a pale
yellow oil, which on storaqe at -20 crystallized as
colorless prisms: mp 62-3C.
lH NHR (CDC13) 1.48 (m, lH), 1.70 (m, lH), 1.79 (s,
3H), 1.89 (s, 3H), 2.05 (m, 2H), 2.65 (m, 2H) and 7.25 (m,
5H).
13C NMR (CDC13) 8.3, 10.6, 24.7, 35.4, 107.2, 125.1,
125.9, 128.4, 141.5, 158.1 and 172.6.
HRM3 m/e: exact mass calculated for C15H1803 (~+)
246.1256, found 246.1270.
15 ~ 11
3.4-Dimethyl-5-(2-phen~lpropyl)-2(5H)-furanone (Compound
8)
Potassium borohydride (503 mg, 9.3 ~mol) was added to
a solution of 3,4-dimethyl-5-hydroxy-5-(2-phenylpropyl)-2-
furanone (Co~pou~ 24, 382 mg, 1.6 mmol) intetrahydrofuran (8 ml) and methanol (6 ml) at room
temperature. After 7 hours, most of the ~olvent was
removed and water (10 ml) was added. Extraction (ethyl
acetate) and evaporation of the dr~ed (m~gnesium sulphate)
extract~ gave ~n oil, which was pur~fied by preparative
tlc ~20x20 cm, 2000u si-lica plate; developed with 30%--
ethyl ether/petroleum ether). The title furanone was
obta~n~d as colorl-~s prisms (recry~tallizsd from ethyl
ether/petrol~um Qthar): mp 69-70.
lH NMR (CDC13) 1.50 tm, lH), 1-.77 (p, 2H, J ~ 6.8
HZ), 1.82 (g, 3H), 1.92 (~, 3H), 1.95 (m,-lH1, 2.67 (~,
2~), 4.74 (m, lH) and 7.25 (~, 5H). :
13C NMR (CDC13) 8.4, 11.9, 26.0, 31.5, 35.4, 82.9,
. ..
. . .
Wo 91/16055 2 ~ 7 3 7 7 ~ PCr/USsl/02005
34
123.6, 125.9, 128.4, 141.5 and 158.9.
HRMS m/e: exact n~ass calculated for C15H1802 (M+)
230.1307, found 230.1311.
E~PD1e 12
4-Octyl-5-hYdroxy-2(5H)-furanone (Co~pou~ 25)
A mixture of glyoxylic acid monohydrate (Co~pou~d 5,
1.19 g, 16.1 mmol), morpholine hydrochloride (1.81 g, 14.6
mmol), water (0.73 ml) and l-decanal (2.89 ml, 15.4 mmol)
in dioxane (6 ml) was stirred at room temperature for 1
hour, followed by reflux for 25 hours. After cooling,
most of the solvent was removed by evaporation and the
residue wa~ extracted with ethyl eth~r. Evaporation of
the dried (magnesium sulfate) extracts qave an oil, which
was flash chromatographed with 30S ethyl acetate/hexane to
give the title furanone.
1H N~ (CDC13): 0.89 (t, 3H, J - 6.6 Hz), 1.25 (br s,
lOH), 1.60 (m, 2H), 2.40 (m, 2H), 4.70 (br, lH), 5.84 (s,
lH) and 6.00 (8, lH).
4-octyl-~methQ~L-fur~ (Coupou~ 26)
A mixture of 4-octyl-5-hydroxy-2(5H)-furanone
(Co~pou~l~ 2S, 244 mg, 1.16 Dmol) and ~-toluenesulfonic
acid (33 mg, 0.17 D~mol) ~nd m~ nol (5.8 ml) was stirred
at room temperature for 2 d~y~. The ~ixture was diluted
with ethyl ether and washed thoroughly wlth 5S sodium
bicarbon~te solution. E~ por~tion of the dried (magnesium
sul~at~) organic phase g~ve ~n oil, which was flash
chromatographed on silica u~ing 10% ethyl ~cet~te/hexane
to gi~- the title fur~none.
lH Nl~ (CDC13): 0.89 (t, 3H, J -- 7.5 Hz), 1.28 (br s,
lOH), 1.60 (~, 2H), 2.35 (m, 2H), 3.56 (8, 3H), 5.64 (8,
lH) ~nd 5.86 (8 lH).
3-Br~?~Q-4-octy~-5-~tl~ox~-2r5H)-~urarlpne (Co~pou~ 27)
A solution of bro~ine (28 mlcroliter) in
- . . : ~ .
:-
WO 91/160~ PCI`/VS9t/02005
2~7a7 7:~
carbontetrachloride (0.2 ml) was added to a solution of 4-
octyl-5-methoxy-2(5H)-furanone (100 mg, 0.45 mmol) in
carbon tetrachloride (0.5 ml) at 0. The mixture was
stirred at room temperature until all the starting
- 5 material ~lsappeared (as monit13Fed by tlc). After cooling
to 0, pyridine (86 microliter, 1.17 mmol) was added. The
mixture was quenched with water and ~he layers were
separated. ~vaporation of the dried (magnesium sulfate)
organic phase gave an oil, which was purified by flash
chromatography using 5~ ethyl acetate/hexane to give the
title furanone.
1H NMR (CDC13): 0.89 (t, 3H, J - 6.8 Hz), 1.28 (br s,
lSH), 1.60 tm, 2H), 2.50 (m, 2H), 3.58 (8, 3H) and 5.69
(s, 1~). \'`
3-Bromo-4-octyl-5-~ydroxy-2L~jHl-furano~ (Co~poun~ 9)
A mixture of 3-bromo-4-octyl-5-methoxy-2(5H)-furanone
(106 mg, 0.35 mmol) and conc~ntrated hydrochloric acid
(0.21 ml) was r~fluxQd unt~l all thQ starting material
disappeared as shown by tlc. After cooli~ng, the mixture
was diluted with ethyl ~ther and wa~ neutral~zed by
washing thoroughly with ~aturatQd potass~um bicarbonate
solution.. Evaporation of the dried (magn~ium sulfate)
organic phase gav~ n oil, which wa~ flash chromatographed
on silica u~ing 10% ethyl acetate/hexane to give the title
furanon~.
IR (C~C13): 3389, 1755 and 1651.
- lH NMR (CDC13): 0.87 (t, 3H, J - 7.2 Hz), 1.27 (br ~,
lOH), 1.60 (m, 2H), 2.49 (t, 2H, J - 8.5 Hz), 4.50 (br,
lH), and 6.05 (~, lH).
13C NMR (CDC13): 14.2, 22.7, 26.5, 27.8, 2g.2, 29.6,
31.8, 98.8, 112.1 and 163.9.
HRNS ~XACt ma~ calculat~d for C12~20~rO3 (M+H)
291.0596, tound 291.0590.
~,.
. . . : . . . :. : . . .. :: . . , . . . . :, : . , ,. .: .: . , : . . . : . . .
