Note: Descriptions are shown in the official language in which they were submitted.
~0 93t20044 PCTJCB93/00708
-1,4 NAPHTHOQUINONE DERIVATIVES WITH ANTI-PROTOZOAL AND ANTI-PARASTIC ACTIVITY
The present inven~ion relates .o naphthoquinones and their use in
chemotherapy. More specifically the invention is concerned ~i.h novel
carbona~e and carbamate deriva~ives ol hydroxynaphthoauinones,
processes for their preparation, pnarmaceutical formulaeions .hereof
and ~heir use in the chemotherapY of certain protozoal and oarasitic
infections.
.
Parasitic ?ro~ozoal infections are responsible for a wide varie~v of
diseases of medical and ve~erinary importance, including malaria in
man and various coccidioses in birds, fish and mammals. .~any o~ the
diseases are life-threa~ening to the hos~ and cause considerable
economic loss in animal husbandry. Parasitic protozoa include the
Apicomplexa, such as species of Eimeria, Cryptosporidium, Toxoplasma,
and Plasmodium. Another parasitic organism of increasing concern is
P~eumocvstis cari~i}, which can cause an often-fatal pneumonia in
immunodeficient or immunocompromised hosts, including those infected
with HIV. The classification of ~his organism is unclear and there is
still unCertaLnty as to whether ic is a protozoan or a fungus.
A wide range or naphthoquinones is ~nown in the art. Such compounds
have been variously described as having antimalarial, anticoccidial
and antitheilerial activity. Ssme compounds have also been described
as possessing activity against external parasites. Thus, Fieser et
al, J. Amer. Chem-.~Soc- L948, 70, 3156-3165 (and rererences cited
therein) describes a large number of 2-substituted-3-hydroxy-1,4-naph-
thoquinones as having antimalarial activity. .~ number of these
compounds have also been described in U.S. Paeent Specification ~o. 2
553 648. Further classes of 2-substituted-3-hydroxy-1,4-naphtho~ui-
nones naving acti~it~ as antimalarial, anticoccidial and/or
.. ..
antitheilerial agents are~described in U.S. Patenes Nos. 3 36/ 830,
and 3 347 742, U.K. Patent Specification No. 1553424, and European
Patent Sp-ciiicaçions Nos. 2 228, 77551, 77550 and 123,238.
W O 93f20~44 .-t~ 't ~ PCT/GB93/007~ -
European Patent .~pplication No. 362996 discloses~ for ehe ;_ea~ment
and/or prophyla~is or infections caused bv Pneumocvs~ls ~3~inl~,
1,4-naphthoquinones of formula:
! I ¦ (I)
_,~ ~,,,\ ,
where Ra is an optionally substituted, Cl 35 non-aromatic hydrocarbon .
residue and R is inter alia a group -OCOR , OR , SR or NReRf, which
compounds are said to be believed to act as pro-drugs of compounds
wherein Rb is a hydroxyl group.
It has now been found tha~ a 1,4-naphthoquinone substituted at the
2-position by a carbamate group exhibits goot acti~ity in v~o ajsainst
malaria in mice infected with Plasmodiu~_yoelii.
Thus, in a first aspect the pre5ent invention provides a compound of
general formula tII)
Rl
R3
' ", ~ (II) I
-' ~/--11 / R2
~ 1
,
wherein
PI~J/68~ 3 1 ~ 0 7 0 ~
- 3 ;~ ' 21 FEBRUARY 1994
~ R3 is Cl 35 hydrocarbyl group optionally substituted by one to five substituents,
selected ~om halo, Cl 6 alkoxy, hydroxy, amino, and mono-or di-Cl 4 alkyl-
amino; and either the dotted line represents a double bond between the 2 and 3
positions of the naphthyl nng; Rl and R4 each represent = 0; and R2 represents agroup
~
'O--C ~R6
A-~CH2)n- N~
R7
wherein n is 2 or 3, R6 and R7 which may be the same or different, each represent
a hydrogen atom, a C1 6 alkyl group, optionally substituted by hydroxy,
O
Cl 6 alko~ or a group C-R9 wherein R9 is a Cl 6 aLkyl group optionally
substituted by an amino group optionally substituted by one or two alkyl groups,or R6 and R7 may each be a group
O
C o-R8 wherein R8 is a C 1-6 aLtcyl group, A is an oxygen atom or
R5
a group N wherein R5 represents a hydrogen atom, a C1 6 allyl group optionally
substituted by hydroxy, C1 6 alkoxy or an amino group optionally substituted by
one or two C 1-6 all~rl groups or R5
O . ,
is a group -C-O-R10 wherein R10- is a Cl 6 ~LIkyl group or R5 is a
Rll
group (CH2) -~ wherein p Is 2 or 3 and Rl 1 and Rl2 are as
hereinbefore defined for R6 and R7 or, when A is -N- and n is 2 R5
may be linked to R6 so tha~ N(CH ~ N forms a piperazine nng;
-
I Pt'1~ 'r , ~ S~
4 f~ 3 ~ O 0 7 0
r~ _ J ~ i 2 l ~EBRUARY 1994
or the dotte~ line represents double bonds between the 1, 2 and 3, 4 positions of thenaphthyl ring, and
R1, R2 and R4 each represent a group
O ~R6
\ O / C \ ~ (CH2)n--N
(wherein n, A, R6 and R7 are as hereinbefore defined); ~'
and physiologically acceptable salts thereo
The Cl 35 hydrocarbyl group R3 may be a straight or branched chain Cl l4 (e.g.
Cl-8)aLlYl or C2-14 (e g- C2-8)a~enYI group or a C3 10 (e.g. C3 g)cycloalkyl
group, each of which may be optionally substituted by a C3 10 (e.g. C3 6)
cycloaL~cyl group, and each of the aforesaid cycloallyl groups optionally bemg
substituted by a Cl lo (e.g. Cl~) alkyl group. Alternatively the hydrocarbyl ' `~
group R3 may be a C3 10 (e.g.C3 8) cycloaLtcyl group substituted by a phenyl
~oup which is optionally substituted by a Cl lo (e.g. Cl 4) alkyl group. The
hydrocarbyl group R3 preferably contains from 1 to 20 carbon atoms, e.g. I to 14carbon atoms. Suitable groups R3 include C3 10 cycloalkyl-CI g-alkyl, Cl lo
alkyl-C3 10 cycloalkyl, Cl loalkyl--C3 10 cycloalkyl-Cl lo alkyl and C3 10-
cycloa~l-C3 10 cycIoalkyl. R3 may also be substituted as hereinbefore defined
by one to five halo, Cl 6 alkoxy, hydroxy, amin~,- Qr mono - or di-C14 alkyl
amino substituents. ~~
PreferredgroupsR3 include:
a Cl 1OaL~cyl group; - -
....
,,, i, .~ . " '1 ~
W O 93/20044 PCT/GB93/0070X
a C5 7 cycloalkyl group (which may be optionally substituted by astraight or branched chain Cl 6 alkyl group, a halo-Cl ,alkyl
group, a Cl 6alkoxy group or a phenyl group, the phen~l group
itself being optionally substitueed by one or ~ore substituents
selected from Cl 6 alkyl and halogen); and
a Cl 1Oalkyl-C5 7cycloalkyl group, wherein the cycloalkyl moiety
maY be optionally su~stituted as defir.ed for the aforementioned
C5 7 Cycloalkyl group.
A particularly preferred class of subscituents R is represented by
the formula:
G 'C~3
!CH2)q~
c~3
wherein: !
q i5 zero or one and -
R is a Cl 10 alkyl group- ~
Another par~icularly preferred class of substl~uents R is represan~ed
by the formula: ~
-(C~2~
wherein:
'i ~l .3 ;~ 7 li~ ~f/~ 9 3 / ~ 0 7 0 ~
21 FEBRUARY 1994
r is zero or one, and either R14 is hydrogen and R~5 is selected from
halo, halo-Cl 6 alkyl, Cl 6 alkoxy, C1 6 alkyl-C1 6 alkoxy, and phenyl
substituted by one or two groups selec~ed from halo and C 1 6 alkyl
or
R14 and R15 are both C1 6 alkyl or phenyl.
.
It will be appreciated that the compounds of formula (I) wherein R3 contains a
substituted cyclohexyl group may e~ist as the cis or trans isomer, th~t is to say
that the cyclohexyl ring may be cls or trans~ substituted by the naphthoguinone
nucleus and the subsistent on the cyclohexyl ring. Both cis and trans isomers and
mixtures thereof in any ratio may be used in accordance with the present
invention. In general when the compound is in the forrn of a mixture of isomers
the trans isomer will be present in an arnount of about 50% or will be the `
predominant isomer but the use of mixtures in which the ~ isomer predominates
is also included within the scope of the invention. The specific ratio of isomers
may be varied as required; typical mixh~es include those in which the cis/trans
isomer ratio is about 1:1,40:60 and 5:9~. For use according to the present
invention the trans isorner of such compounds of formula (I), or a mixture of the
cis and trans isomers containing at least 95% e.g. 99% of the trans isomer, is
preferred.
An especially preferred substituent R3 is the 4-(4-chlorophenyl3cyclo-
hexyl group:
~ y \\
in particular in the trans forrn with respect to the naphthoquinone ring. I
~J ~ ; ' , r ; i ~ S
P'`T ~ " ~ t ~,i~ ; ~~~~ ' ` ' ~ I -
~? ~
W O 93/20044 ~ i PCT/GB93/00708
n the compounds or rormula (II) .~ is preferably a group _~T_ and R
represents a carbamate group
O
, (CH ) 6
\o/ \ N/ \ N/ R
\
R5 R7
y of the groups R , R , R , R8, R10 Rl1 or R
alkyl, this may be straight or branched chain e.g. methyl, ethyl,
isopropyl, t-butyl, isopentyl or n-hexyl. Suieably R, R and R are
each hydrogen or a Cl 4 alkyl group. Most suit~bly R5 is methyl or
- ethyl, R6 is hydrogen, methyl or ethyl, R ic hydrogen or a group
-C0-0-R wherein R is Cl 4 alkyl, e.g. t-butyl.
In the compounds of formula (II) R and R preferably represen~ -O and
-.he dotted line is a bond between the 2 and 3 positions of the
naphehyl ring.
A preferred group of compounds of formula (II) may-be-represented by
.he formula (III) ~ - -~ j
G~\/~j /--~ R6
j C~Z)n N
~herein R , R and R are as hereinbefore defined,
and physiolo~ically acceptable sales ~hereof.
W O 93/~'0044 , ~ 8 - PCT/GB93/0070~
~ ~urthermore ~he compounds of formula ~III) are prererably in the form
of the ~rans-isomer.
''-[t-ans-4-(4-Chlorophenvl)cyclohexyl]-3-lN-methyl-~-;2-(methylamino)-
ethyl]arbamoyloxv~ -naphthoquinone is a preferred compound of ~he
formula (III).
A fur~her preferred group of compounds of formula (II) may be
represen~ed by ,he formula (IV~:
9 ,~ (IV)
~/ \ I ~ Q R
. ~ ~ J ~ ¦ ~6
`(CH2)n N
wherein R5, R6 and R are as hereinbefore defined
2-~trans-~4-~-butylcyclohexyl)methyl]-3-~N-meehyl-N-[2-(meehylamino)-
ethyl]carbæmoyloxy)-1,4-naphthoquinone is a preferred compound of the
formula (IV). _ -
By the term "hydrocarbyl" group is meant an aliphatic group, e.g. astraight branched chai~-or-cyc:l-ic alkyl, alkenyl or alkynyl group, a
carbocyclic aryl, an aliphatic group substituted by a carbocyclic aryl
group optionally substituted by an aliphatic group or a carbocyclic
aryl group substituted by-an aliphatic group.
Without wishing -o be bound by theory, it is believed that the
carbonate or carbamate ;derivatives of formula (II) are pro-drugs of
the corresponding hydroxvnaphthoquinone, that is, the carbamate or
carbonate or group is cleaved in vivo to give the compound of formula
(I) wherein Rb is a hydroxyl group.
!
~1 -f ~ `-~ rl ~ '
~ ~ I j J ~ L ! '
WO 93/20044 PCr/(~B93/0070~ .`
q
It is believed that the compounds of formuia (II) ~ xhibit
activity against parasi~ic protozoa, in particular those organisms
against which corresponding hydroxvnaphthoquinones have been found to
be acti~e. such as Plasmodium species, eg. P. alci~arum; ~imeria
s?ecies eg. _.~enella and E.ace~lina; rheileria species a.g
T parvum and T annulata; Cr~tos~oridium; and To~oolasma ~ondii as
well as .he parasitic organism Pnewmocvstis carinii, and ~ill
.herefore be useîul in the ereatmene and/or prophylaxis of parasitic
infections, such as those caused bv parasitic protozoa, eg. malaria,
coccidiosis, cryptosporidiosis, toxoplasmosis and those caused by
~_sL~i~iL eg. P.carinii pneumonia (PCP) in animals, including humans.
It will be appreciated that the amount of a compound of formula (II)
or its salt required for use in the treatment or prophylaxis of the
above-mentioned diseases will depend inter ~lia on ~he particular
compound administered, the route of administration, the age and weight
of the mammal ~e.g. human) to be treated and the nature and se~erity
of the condition being treated. In general, a suitable dose for
administra~ion to man for the treatment of malaria is in the range of
O.lmg to 200mg per kilogram bodyweight per day, for example from
lmg/kg .o lOOmg/kg, particularly 10 to 40 mg/kg. It will be
apprecia~ed that for administration to neonates, lower doses may be
required. ~ ~ -
For prophylactic treatment a compound of formula (II) Qr a saltthereof may also be given less fre~uently, e.g. as a-s mgla dose on
alternate days, once or t~ice per week or once or twice per month.
The dosage for prophylatic treatment will depend inter alia on the
frequency of administration, and, where a depot preparation or
controlled release formula~ion is used the raee of release of the
active ingredient. Thus for once-weekly administracion a suitable
prophylactic dose is in che range 0.1 to 100 mg/k~,e.g. O.S ~o 50
mg/kg par~icularly 5 to 50 mgjkg.
W O ~3/20044 ~ 7 /~ ! PCT/GB93/0070~
=
'. snouid be understood ~hat for consistency the dosages referred to
above are calculated in terms of ~he compound of formula (II) ~ se,
and mav ~equire adjustmene in the evenc a salt is emploved.
Ihe present invention ~hus rurther 2rovides a method for the treatment
and/or propnylaxis of parasieic inrections e.g. parasitic protozoal
infections such as malaria, or infections caused ~y P.carinii, in
mammals e.g. humans, which comprises administering to a mammal
suffering from or susceptible to said infection, an effective amount
of a compound of formula (II) or a physiologically acceptable salt
.hereof.
There is also pro~ided a compound of formula (II) or a physiologically
acceptable salt thereof for use in therapy, e.g. in the treatment
and~or prophylaxis of parasitic diseases as hereinbefore defined.
!
The invention also provides the use of a compound of formula (II) or a
physiologically acceptable salt thereof for the manufacture of a
medicament for the treatment and~or prophylaxis of parasitic
infections as hereinbefore defined. '
For use according to the present in~ention a compound of formula tII) -
or a ?hysiologically acceptable-~salt thereof is preferably presented
as a pharmaceutical formulation. In general such pharmaceutical
formulations will comprise - ? compound of formula (Il) or a
physiologically acceptable sait thereof together with one or more
pharmaceutically acceptable carriers therefor and optionally other
~herape~tic and/or prophylactic ingredients. The carrier(s) must be
acceptable in the sense o ~eing compatible with the other ingredients
of the formula and not de-Leeerious to the recipient thereof, .
:,
The present invention, -therefore, further provides a pharmaceutical
'ormula~ion comprising a compound of formula ~II) or a pharmaceuti-
cally acceptable sar; ehereof cogeeher with a pharmaceutically
acceveable carrier ~herefor.
' '' 1 !" . ~
W O 93~20044 PCT/GB93/00708
There is aiso provided a method ror the preparation of a pharmaceut-
ical for~ulation comprising bringing into associazion a compound o~
~ormula (I ) or a pharmaceu~ically acceptable salt thereof, and a
?harmaceutically acceptable carrier therefor.
.~ co~oound of formuia (II) or ies salt may conveniently be presen~edas a pharmaceuticai formulation in unic dosage form. A convenient
unit dose formulation contains a compound of formuia (Il) or a
physiologicallv acceptable salt thereof in an amount of from 10 mg ~o
lg.
Iharmaceu~ical formulations include those suitable for oral, _oplcal
~including dermal, buccal and sublingual), rectal ~nd parenteral
(including subcutaneous, intradermal. intramuscular and intravenous)
administration. The formulation may, where appropriate, be
convenienrly presen~ed in discrete dosage units and may be prepared by
any of the methods well known in the art of pharmacy. All methods
incLude the step of bringing into association a compound of formula j
(II) or a physiologically acceptable salt thereof with liquid carriers
or finel,v dividec solid carriers or both and then, if necessary,
shaping ~he product into the desired formulation.
,
Pharmaceutical formulations suitable for oral administration wherein
the carr-er is a solid are most preferably presented as unit dose
formulations such as boluses, capsules or tablets each containing a
predetermined amount of the active ingredient(s). A tab-let--m~y be
made by compression or moulding, op~ionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing in a
suitable machine the ac~ive ingredient(s) in a free-flowin~ form such
as a powder or granules optionally mixed with a binder, -lubricant,
inert diluent, lubricating agent, surface-acrive agent or dLspersing
agen~. Moulded tablets may be made by moulding in a suitaUe machine
a mi~rure of the powdered active ingrediene(s) with any suitable
carrier. Table~s may be optionallv coated and, if uncoated, ~ay
optionally be scored. Capsules may be prepared by filling the active
W O 93/20044 i! ' "! ~ -1 ,1 ,, PCT/GB93/0070f
,_, , ),, ~, 1 1
ingredien~(s) , either alone or in aomixture ~ith one or ~ore
accessory ingredients. ineo the capsule sh~lls and .hen seaiing he~
in the usual manner. Cachets are analogous ~o capsules wherein ~he
active ingredien~(s) ~ogether wi.h anv accessorv ingredient(s) is
(~re) seaied ln a rice paper envelo~e. .~ co~pound or formula (II) or
a physiologically accepta~le salt .hereof ~av also be rormulated as
dispersible granules, ~hich may for example be suspended in ~ater
before administration, or sprinkled on food. The granules mav be
packaged e.g. in a sache~. Formulations suitable for oral
administration wherein the carrier is a liquid may be presented as a
solution or a suspension in an aqueous lîquid or a non-aqueous liquid,
or as an oil-in-wa~er liquid emulsion, e.g. a sYrup, elixir, emuision
or a draught. A syrup may be made by adding the active ingredien~(s)
eo a concentrated aqueous solution of a sugar, for example sucrose, to
which may also be added any accessory ingredients which may include
flavourings, agents to retard crystallisation of the sugar or agents
which increase the solubility of oeher ingredients such as polyhvdric
alcohols for example glycerol or sorbitol.
Formulations for oral administration include controlled release dosage
forms e.g. tablets wherein the ac~ive ingredient(s) is (are)
formulated in an appropriate-release - controlling matri~, or coated
with a suitable release -~con~rollIng film. Such formulations mav be
particularly convenient for prophylactic use.
F
Pharmaceueical formulations suitable-for rectal administration wherein
the carrier is a solid are most preferably presented as unit dose
suppositories. Suitable carriers include cocoa butter and other
materials commonly used in- the art. The suppositories may be
conveniently formed bY admixture of the active ingredient(s) with the
softened or melted carrier(s)- followed by chilling and shaping in
moulds. -----
Pharmaceutical formulati-ons suitable for parenteral administration
include sterile solutions or suspensions of the active ingredient(s)
? ~ -'? 7,1 '
W O 93~20044 ~ ' PCT/GB93/00708
- 13 -
in aaueous or oleaginous vehicles. Injec~ible preparations may be
adapted for bolus injec~ion or continuou~ infusion. Such preparations
are con~eniently presented in ~nit dose or multi-dose containers
which are sealed after introduction of the formula~ion until requ~red
for use. .~lternatively, the active ingredient(s) may be in powder
form eg. freeze-dried which is constituted with a s~litable vehicle,
such as s~erile, pyrogen-free water, before use.
A compound oî formula (II) or a physiologicallv acceptable salt
thereof mav also be formulated as a long-acting depot preparation,
which mav be administered by intramuscular injection or by
implanta~ion e.g. subcutaneously or intramuscularly.~epoe preparations
may include, for example, suitable polymeric or hydrophobic materials,
or ion-e~change resins. Such long-acting formulations are
particularly ~onvenient for prophylactic use.
I
Uhilst the compounds of the invention may be formulated in any known
manner, for example as described above, in view of their properties
they are particularly suited for formulation, for example as aqueous
solutions which may be used for injection. 2-[erans-4-(4-chlorophen
yl)cyclohexyl~-3-~N-methyl-N-[2-(methylamino)ethyllcarbamyloxy)-1,4-
naphthoq~inone has improved water solubility over the parent
hydroxynaphtho~uinone. ~ ~
It should be understood that in addition to the aforemen~ioned carrier
ingretien~s the pharmaceutical formulations for the various routes o~
administration described above may include, as appropriate one or more
additional carrier ingredients such as diluen~s, buffers, flavouring
agents, binders, surface active agents, thickeners, lubricanti,
preser~atives (including an~i-oxidants) and ~he like, and substances
included for the purpose of rendering the formulation iso~onic _~th
the bloot of the intended recipient. -~ ~
The compounds of the present invention may be administered in
combination or concurren~ly wi~h other ~herapeu~ic agen~s, for example
W 0 93t20044 ~ PCT/GB93/~070
other antimalarial agen~s, such as 4-aminoquinolines eg. chloroquine
and amodiaauine; 8-aminoquino- lines eg. primaquine; chloroquine;
mefloquine; quinine; quinidine; artemesinin; ar~esumate; ar~emether; ~ ;
haloîantrine; dihydrofolate reductase inhibi~ors eg. pyrime~hamine; r.~
sulphonamides eg. sulphadox- ine: proguanil; chloroproguanil;
dapsone, hydroxvnaphthoquinones: or ~ixtures eg.
pyrimethamine/sulphadoxine; pyrime~hamine/dapsone; and ' ;
pyrime~hamine/sulfalene; antibac~erial agents such as trimethoprim-
sulphamethoxazole mixtures; an~icoccidial agents such as monensin,
ha}ofuginone, arprinocid. amprolium. dinitolmîde, robenidine or
salinomycin; or antibio~ics such as clindamycin, tetracycline, ¦-
doxvcycline, or spiromycin; or agen~s active against Pneumocvstis
carinii sucn as pen~amidine or Eflornithine.
~hen compounts of formula (II) are used in combina~ion with a second
therapeutic agent the dose of each compound may vary from that
required when the compound is used alone. Appropriate dosages can be
readily determined by those skilled in the ar~. ¦
Compounds of formula (II) may be prepared by reaction of a compound of
formula (V)
~ ~3
''~/\ ~ (V)
O ..
wherein R3 is as hereinbelore defined and-X is halo, with a compound
of the formula (Vl)
HA(C~)n N\ (VI)
R
? ~ 1 r/ ,1,
W O 93/20044 PCT/GB93/00708
- 15 -
wherein n, .~, R and R are as nereinbefore defined; 2rovided ~hat
when it is required to prepare a compound wherein one or more of R
and R is hvdrogen. the corresponding compound is prepared with a
group -ÇO-O-R in place, instead or hydrogen and this group is ~hen
removed.
The reaction may con~eniently be effecced in the presence of a solvent
which is inert tO the reagents. Solvents which may be emploved
include aromatic hydrocarbons e.g. benzene or toluene; halogenated
hydrocarbon e.g. chloroform or dichloromethane; dipolar apro~ic
soivents e.g. dimethylformamide or hexamethylphosphoric ~riamide;
e~hers e.g. tetrahydrofuran or dioxan; pyridine; acetonitrile;
trimethylphospha~e and triethylphosphate. The reaction temperature
may conveniently be in the range of from -80 C tO 100 C, preferably
0C to 30C.
The reaction may adYantageously be effected in the presence of a base
which may be an organic base, for example pyridine, 4-dimethylamino-
pyridine, a terti~ry amine such as .riethylamine, or l,8-diazabicy-
clo~5,4,0]-7-undecene (D W), or an inorganic base, for example an
alkali metal carbonate such as potassium carbonate or sodium hydrogen
carbonate.
- To prepare a compou~d of-formula (II) wherein Rl, R2 and R4 each
represent a carbonate- or- carbamate group che reaction should be
effected after reduction of the quinone nucleus, which mav be effected
in conventional manner.
Sale formation can also be effected by methods well known in the art, I
by reacting a c~mpoun~ of formula (II) with an appropria~e acid.
. _
RemoYal of the pro~ecting ~roup -C0-0-R is conveniently carried out
by acid, for example when R is tertiary butyl hydrogen chloride in
ether.
W O 93/20044
- o
The compounds or ~he rormula (V) mav convenien~l~ be prepared ~n situ
from the corresponding hvdroxvnaph~hoquinone, ror example when .~ is
chloro bv the reaction oI the hydroxynaph~hoquinone wi~h phosgene in
an inert soi~ent, as defined above, ln the presence of the base used
.o actach ~he carbama~e side chain. This reaction is carried ou~
between -~0 C and 50 C and suitably ~t between -10 C and 10 C.
5Ompounds of formula ~IV) may be prepared according to known ~ethods
for the svnthesis or hydroxynaphthoquinone deriva~ives, such as are
described in US Parents ~os. 2,553,648; 3,367,830; and 3,347,742; UK
Paten~ No. 1,553,424; European Patents Nos. 2,228; 77,551; '7,550; and
123,238 and European Patent Applica~ion No. 362,996. Thus for example
a compound of formula (IV) may be prepared by reaction or a 2-halo
(e.g. 2-chloro)-1,4-naphthoquinone with a compound serving tO include
ehe required R group, followed by alkaline hydrolysis. Where R is
an optionally substituted cyclohexyl or optionally substituted
cyclohexylmethyl group it may be in~roduced by reaction with the
appropriately substituted cyclohexylcarboxylic acid or
cyclohexylacetic acid.
The i~vention will now be further illustrated by the following non-
limiting examples:-
In vi~o Activitv a~ain~t ~lasmodium ~elii_in mice.
Tes~ Com~ounds
Compound A: 2-[trans-4-(4-Chlorophenyl)cyclohexyl]-3-~N-methyl-N-
~ ~2-(methylamino)ethyl]carbamoyloxy)-1,4-naphthoquinone hydrochloride
mo- nohydrate. I
_
Com~ound B: 2-[trans-4-(4-Chlorophenyl)cyclohexyl ! - 3-lN-[2-(methvl
amino)ethyl~car~amoYloxy~-1,4-naph~hoquinone hydrochloride monohy-
drate.
.~,t ~,.-7 ,~ ,
W O 93/20044 PCT/GB93/00708
l7 ,
Co~ound C: 2-[trans-4-(4-Chlorophenyl~cyclohexylj-3-~N-me~hvl-
~'3-(me~hvlamino)propyl]carbamoyloxy)-1,4-naphthoquinone
Com~ound _D- trans-2-~4-(4-Chlorophenyl)cyclohexyl]-3-~piperazine-N-
carbonyloxyj-1,4-naphthoquinone hydrochloride dihydra~e,
Com~ound (1~: 2-[trans-4-(4-Chlorophenyl)cyclohexyl]-3-hydrox~-1,4-
naphthoquinone.
Compounds A, B, C, and (I) were administered orally and intravenously,
in the manner described below,
METHOD
I
The test method is a modified version of the 4-day suppressive tes~.
The YM strain of P,voelii uas maineained in mice by twice weekly
passage. Infected blood was collected and diluted with normal saline
to give an inoculum of 3X106 parasitized enythrocytes/ml. On the
morning of Day 1 the test animals (male CD-l mice weighing 18-20g)
were infected intravenously via the tail vein with 0,lml of
inoculum~mouse.
I
Oral (p.o.)
Test compounds A, B and ~ -were formulated by ball milling in 0.25%
celacol with stainless steel balls. The total requirement was
formulated at the beginning of a test and thereafter stored at 4 C.
.
Each test compound was administered orally (p.o,) to four or more
usually fi~e groups of mice, each group recei~ing a different dose :
level (dilution) _ A-- further group of mice was administered celacol
only, as a control- ~ie, a total of 30 mice are used to obtain each
ED50). The mice were dosed orally 24 hours post-infection.
Intravenous (~,v ~
WO 93/20044 ;1 ~ r~ ~; r~ 18 - PCT/G~93/0070~
I.-. formuiations of .es~ Compounds .~ and B were pre~ared by
dissolving the compound in wa~er lmmediately berore use, and
administered via the .ail vein as a single dose 24 hours
post-infection.
On ~he morning of day ~ smears ot tail blood were prepared from each
mouse and the parasitaemia coun~ed. 3ata were analysed to yield EDj~
values bv the best fit to a sigmoidal dose response curve. The
results are presented in Table 1 below.
Results
Table 1
Dosing Schedule
ED50 mg/kg
I ~: P.o. 1 ~ i.v,
I
Com~ound (I~ 0.16 0.12
~- Compound (A) O.15 0.12
- Co ound (B) 0.2 0.27
.
.,
Com~ound (C~ O.4 0.29
~ ~ Com~u~ (D) O.4
-
In vi~o Act vit~ a~ainst Pneumoc~stis carinii in mice
.
Dru~s .ested
~ Septrin - Wellcome Paediatric Suspension con~aining 200 mg
sulfamethoxazole/40 mg trimethoprim per 5 ml
'~ ?~
W O 93/20044 PCT/GB93/00,08
l9
~tovaquone-2- .rans-4- (4-chlorophenyl)cyclonexyl]-3-hydroxy-1,
4-naphthoquinone
Test compound-2-[trans-4-(4-chlorophenyl)cvclohexyl~-3-~N-me~hyl-
N- E 2-(methylamino)e~hyl~ carbamovloxy)-1,4-naphtho-
quinone hydrochloride monohvdrate
~ethod
?neumocvs~is_carinii (PC~-free female SCID (C.B-i7/lcr-scid/scid) mice
(20-25g) were started on dexamethasone (2m/L in drinking water), 7
days prior to in~ratracheal infection wi~h single dose of
cryo-preserved mouse-derived PC (dose:-2 x lO cysrs equivalent to
about 106 .rophozoites). Drugs were evaluated for prophylaxis bv
continuous daily oral dosing of groups of 10 mice wieh each drug from
1 dav post infection until day 42. Trials were terminated and all the
mice killed 24 n after the last dose. The lungs were removed and an
impression smear prepared ~from a representative portion of lung
; - selected a~ random. Lung smears were stained for P.carinii cysts
usin~ an immunofluorescent antibody test kit ~Detect IF P. carinii,
Shield Diagnostics Ltd.) Slites were scored for the intensity of P.
carinii infection by scanning the entire smear a~ lOOx magnification
and coun~ing the highly fluorescent cysts. Smears were assigned one
of the following scores~.
0 - no cysts/smear
+1 ~ 1- to 5 ~ys~s/smear
,
+2 - 6 to 50 cysts/smear
+3 51 to 250 cysts/smear
+4 ~250 cvsts/smear
Drug formulation~
Septrin was diluted int~ ~-drinking water; the dose was calculated on
the basis that each mouse would drin~ a minimum of 2.5 ml/day.
Atovaquone was administered as a suspension in 0.25% celacol. The
test compound was dissolved in distilled water and the solution
administered within L hour of preparation.
WO 93/20044 ~ r~ ~ . PCT/GB93/0070
2 0
~esul tS
Ihe results v~hich are presen~ed in Table 2 below, were based ~n two
independen~ blind examinations, and are expressed as an a~erage
infection score oî the 10 mice in each group. Data are also expressed '
as a percen~age of the control (dosed daily p.o. wl~h dis~illed
water).
TABLE 2
Treatment GroupMean score'+ SE % of Control
Control 3.6+ 0.2100
Septrin (2S0/50 mg/kg/day) 0.1+ 0.1 3~
A~ovaquone ~100 mg/kg/dav p.o) 1.5+ 0.3 43%
Test,compound (100 mg.kg.day p.o) 0.1+ 0.1 3%
FormLulation ~xamPLes
The following examples i}lustrate, with particular reference to ehe
compound of Example 2, pharmaceutical formulations which may be
employed in accordance with the present invention. It will be
appreciated that other compounds of form~la (II) may be formulaced in
similar manner-
.
. Iniectable solut-on
A solution for intramuscular injection may be prepared by
~ ixing:-
=,_~- , Compound of Example 29.5 par~s by weight
~~ Dimethyl sulphoxide19.0 parts by weight
Sorbitan monooleace4.5 parts by weight
Corn oil 67 0 parts by weight
.100 . O
W O 93/20044 ,~ iPCT/GB93J0070
- 21 -
B. Injec~able solution
Compound of Example 2 5 parts by weigh~ .
N-me~hvl-pyrollidone 48.3 parts by weight
Tween 80 2 parts by weight
Span 80 4.7 parts by weight
Miglyol 812 4~l parts by weight
100 0
C. Table~
Compound of Example 2 25.0 mg
Lactose BP 48.5 mg
Microcrystalline Cellulose BP 10.0 mg
("Avicel pH 101")
Low-substituted Hydroxypropyl; 10.0 mg
Cellulose BP (~LHPC LH-ll")
Sodi~m Starch Glycollate BP 3.0 mg
("Explotab")
~:~ Povidone BP (~K30") 3.0 mg
Magnesium Stearate BP 0.5 mg
_ - 100 0 mg
D. IV Solution Freeze Dried
- - j
Compound of Example 2 50mg
Water ~or Inj ections to lOml
Dissolve the naphtho~uinone in the ~ater for Injections.
Sterilise by fi~tration. Fill into glass vials and freeze dry.
Recons~itute-:shortly before use with Water for Injec~ions,
WO 93/20044 ~ ~ I q ' PCT/GB~3/00708
~ _ rj rJ i . ~ - 2 2
E. ~a~sule
Compound of Example 2 100 mg
Starch 1500 150 mg
~agnesium stearate 2.5 mg
filled into a hard gelatin capsule
F. Aerosol Formula~ions
a) Compound of Example 2, micronised 1.0 mg
Aerosol propellant to 5.0 ml
Suspend the micronised campound in ~he aerosol propellan~. ~
- Fill this suspension ineo preformed aerosol cannisters, Sj ~-
ml~cannister under pressure, through the valve orifice.I ~
`~
b) Compound of Example 2, micronised 1.0 mg
Arlacel 85 Q.l~ w/v
Aerosol propellane to 5 ml
!
Disperse the Arlacel 85 in the aerosol propellant and then
add compound of Example 4. Fill the _suspension into
preformed aerosol cannisters, Sml/cannis~er under pressure,
through ~he valve orifice.
: ~~ ~~ G. Powder Inhalation
; Compound of Example 2, micronised 1.0 mg
Lactose 29.0 mg
': ' .
Tritura~e and blend the micronised compound with the lactose.
~~ F~ll ehe resuleing powder blend into hard gelatin capsule shells,
30 mg per capsule.
tert-butvl ~-methvl-~-F2-(me~hvlamino)et~v11carbamate
O 93/20044 PCT/~B93/00708
- ~3 -
This was ?re~ared from N,N'-dimethvlethylenediamine rollowing rhe
procedure of W.S. Saari. J.E. Schwering, P..~. Lyle, S.J. Smith and
E.T. Engeihardt. J.~ed.Chem. vol 33, 97, L990.
~is_'2-~ r~-butoxvcarbonvl~Laminole~hvllamine
To a stirred solu~ion or diethylenetriamine (lOg; 97 ml; Lancaster
synehesis) ~n tetrahydrofuran (5ml) cooled to O C was added dropwise a
solution of di-tert-bu~yl dicarbonate (10.5g; 48mM) in tetrahydrofuran
(50 ml). The mixture was allowed to warm ~o room temperature and
stirred for 18 hours. filtered concentrated Ln vaeuo snd partitioned
be~ween brine and ethyl acetate. The aqueous layer was extrac~ed once
with ethYl acetate and the combined extracts were dried (MgS04) and
concentra~ed Ln vacuo to leave a clear 3il (6.7g). Chomatography on
silica gel with 1:4 methanol: chloroform gave the title compound as a
clear oil (2g; 27%), NMR ~H (CDC13) 3.15-3-3 (4H, m), 2.75 (4H
t,J5Hz), 1.~5 (18 H,s).
Exam~le 1
2- r trans-~-t~-~hloro~hen~l)c~clohex~ 3-~N-me~hYl-~-~2-(N-methYl-
~-tert-butoxvcarbonvl2_minoethyllcarbamovloxYl-1.4-nashtho~uinone
To a stirred suspension of 2-hydroxy-3-ttrans-4-(4-chlorophenyl)cyclo-
xy1]-1,4-naphthoquinone (3.~g;-lOmmol) in dry dichloromethane (60ml),
cooled to O C under nitrogen, -was added all at once a solution of
phosgene in toluene (lOml;- 1.25M solution). Pyridine (0.8ml; lOmmol)
was added dropwise and the solution stirred for a further 45 minutes
at 0C. .~ mixture of~` ~ -butyl N-methyl-N-[2-(methYlamino)echyl~
carbamate (1.85g; -10.5mmol) and pyridine (0.8ml; lOmmol) in dry
dichlorome~hane (lOm}.~ was atded dropwise o~er 15 minutes and the
mixture lert stirring-a-t roo~ temperature for 18 hours. The mixture
was diluted with dichloromethane (50 ml), washed with water, lM
hydrochloric acid and water, dried (magnesium sulphate) and
concentrated ;o lea~e an orange oil. Chromatography on silica with
W 0 93t20044 ~ ' PCT/GB93/0070~ -
1:9 ethyl aceta~e:dichlorome~hane followed bv tr.turation with 1:2
e~her:hexane gave ~he title compo~nd (3.12g; ~4%), ~.p. 113-115 C, NMR
~H (CDC13) 3.0-8.2 (2H, m), 7.65-/.8 (2H. m)~ '.L-7.3 (4H, m), 3.4-3.7
(4H, m), '.'5 (3H, 2~s), ~.05-3.25 (1~. m), 3.0 (3H, 2~s), 1.5-2.15 '`
(9H, 2xs).
Exam~le 2
2-~trans~ -Chloro~nenvl)cvclohex~11-3-~N-methvl N-~2-l~ethylamino)
ethvllcarDamovloxY~-1.4-na~h~ho~uinone hvdrochloride monohvdrate
!
To ether sa~ura~ed with HCL (30ml) was added 2-[crans-4-(4-Chlorophen-
yl)cvclohexvl]-3-(N-methyl-N-~2-(N-methyl-N-tert-butoxvcarbonyl)amino ~.,
ethyl]carbamoyloxy~-1,4-naphthoquinone ~1.74g, 3mmol) and the solution
s~irred at room temperature for 5 hours. The precipi~a~e was filtered
off, wasned with ether and dried in vacuQ to give ~he title compound
(1.48g; 92%), m.p. >140 C tdecomposes). ~MR ~H (CDC13) 9.1-9.3 (lH,
bs), 7.95-8.2 (2H, m),7.65-7.85 (2H, m), 7.1-7.35 (4H, s), 3.7-4.0
~lH, br s), 3.0-3.5 (6H, m), 2.8 (3H, br s), 2.65 (lH, m), 1.75-2.15
(8H, m), 1.~-1.75 (2H, m).
The following were prepared in a similar manner ~o example 1:
2-[trans-4-(4-Chlorophenyl)cyclohexylJ-3-~N-[2-(N-methyl-~-ter~-butoxy
carbonylamino)ethyl~carbamoyloxy)-1,4-naphthoquinone, from tert-butyl
N-me~hyl-N-(2-aminoethyl)car~ama~e (W.S. Saari et al. J.Med.Chem. vol
33, 97, 1990), m.p. 128-131 C, NMR ~H (CDC13) 8.05-8.15 (2H, m),
7.65-7.8 (2H, m), 7.15-7.3 (4H, m), 3.5 (4H, m), 3.0-3.2 (lH, m), 3.0
(3H, s), 2.~-2.7 (lH, m), 1.75-2.15 (6H, m), 1.45-1.7 (llH, m).
,, -
- = 2-[trans--(4-Chlorophenyl)cyclohexyl]-3-(N-methyl-N-[3-(N-meehyl-N-
- _ tert-butoxvcar~onyla~ino)propyl]carbamoyloxy)-1,4-naphehoquinone, from
tert-butyl N-methyl-N-[3-(methylamino)propyl]carbama~e (W.S. Saari e~
_ al. J.Med.Chem. vol 33, 97, 1990), oil, NMR SH (C~C13) 8.0-8.2 (2H,
=), 7.65-;.8 (ZU~ m~, 7.1-7.35 (4U, m), 3.25-3.6 (4U, =~, 3.0-3.25
~ , 7 1 ~
W O 93/20044PCT/GB93/00708
- 25 -
(4H, m), 2.? (3H~ s), ~.5-2.7 (lH, m), 1.75-2.15 (8H, m), 1.'1-1.7
(llH, m).
',~
2-~trans-4-(4-Chlorophenvl)cyclohexyl]-3-(N-ethvl-~-[2-(N-e~hyl-N-
tert-butoxycarbonylamino)ethyl~carbamoyloxy~ napAthoquinone, from
tert-butyl ~-ethyl-N-[2-(ethylamino)ethyl]carbama~e (W.S. Saari ~t al.
J.Med Chem. vol 33, 97, 1990), oil.
2-[trans-4-(4-Chlorophenvl)cyclohexyl]-3-~N-tert-butoxy-carbonylpiper-
azine-N-carbonyloxyj-l,'l-naphthoq~inone, from N-tert-butoxycarbonyl- ¦
piperazine (Carpino et al J.Or~.Chem., vol 48(5), 664, lg83), oil.
2-[erans-4-(4-Chlorophenyl)cyclohexyl~-3-l[2-(N-methyl-N-tert-bu~oxy-
carbonyl amino)ethoxy]carbonyloxy~-1,4-naphthoq~inone, from tert-butyl
N-(2-hydroxyethyl)-N-methylcarbonate ~W.S. Saari et al. J.Med.Chem.
vol 33, 97, 1990), m.p. 110-lll C, NMR ~H (d6-DMSO) 7.9-8.1 (4H, m~,
7.3 (4H, s), 4.35-4.45 (2H, t, J-4H~)), 3.5-3.6 (2H, t, J-4Hz),
3.0-3.15 (lH, m), 2.5-2.7 (IH, m), 1.7-2.0 (6H, m), 1.45-1.65 (2H, m),
1.4~(9H, s).
2-ltrans-4-(4-Chlorophenyl)cyclohexyl]-3-~N,N.-bis~2-(tert-butoxy ~`
carbonylamino)ethyl]~carbamoyloxy~-1,4~naphthoquinone, from bis
~2-[(tert-butoxycarbonyi)aminolethyl~ amine, yellow oil, NMR ~H
(CDC13) 8.0-8.2 (2H,m), 7.65-7.8 (2H, m), 7.1-7.3 ~4H, m), 3.'5-3.7
(4H,m), 3.15(1H, m), 2.62~ , m), 1.8-2.1 (4H, m), 1.45 (9H,br s),
1.4(9H,br s) -- -
, .
The following were prepared in a similar manner tO example 2 from the
corresponding tert-buto~ycarbonyl pro~ected carbonates or carbamates :
,..................................................... ~;.
2-~,trans-4-(4-Ch,lorQphènyl)cyclohexyl]-3-~N-[2-(methylamino)ethyl)car-
bamoyloxy~-1,4-nap~~`t~oq~inone hydrochloride monohydrate, m.p.
194-197 C, ~MR ~H (d6-DMSO) 9.1-9.3 (2H, bs), 8.4-8.5 (lH, t),7.8-8.1
(4H, m), 7.3 (4H, s), 3.4-3.6 (2H, m), 3.0-3.2 (3H, m), 2.55-2.75 (4H,
m), 1.4-Z.15 (8H, m).
W O ~3/20044 PCT~GB93/0070f
,-,.~.~ r)~t ~ - 26 -
, . _ . ~ ~
2-~trans~ 4-Chloropnenyl)cyclohe~yl3-3-lN-me~hvl-N-[3-(~ethylamino)
propyl] carbamoYloxYl-1,4-naphthoq~inone hvdroc~loride monohvdrate,
m.p. 208-210 C, NMR ~H (CDCl3) 8 1-8.2 (lH, d, J~5Hz), 8.0-8.1 (l~, d,
i~SHz), 7.5;-~.85 (2H, m), 7.15-7.3 (4H, m), 3.05-3.35 (7H, m),
2.;-2.8 (5H. m), 1.45-2.1 (lOH, m).
2-~trans-4-~4-Chlorophenyl)cyclohexyl]-3-~N-ethvl-N-~2-(e~hylamino)
ethvl]carbamovloxy)-1,4-n~phthoquinone hydrochloride ~onohvdrate, m.p.
197-201 C. NMR SH (CDC13) 8.1-8.2 (lH, d, JJ8Hz), 8.0-8.1 (lH, ~, -
J~8Hz), 7.65-7.8 (2H, m), 7.1-7.3 (4H, m), 3.0-4.1 (7H, m), 2.5-2.7
(lH, m), 1.25-2.15 (14H, m).
2-[erans-4-(4-Chlorophenyl)cyclohexyl]-3-[piperazine-N-carbonyloxy]-
1,4-naph~hoquinone hydrochloride dihydrate,-m.p. ?160 C (decomposes),
NMR ~H (d6 DMSO) 7.8-8.1 (4H, m), 7.25-7.4 (4H, m), 3.6-4.0 (4H, m),
3.0-3.5 (5H, m), 2.5-2.7 (lH, m), 1.7-2.05 (6H, m), 1 4-1.7 (2H, m). ~,
2-[trans-4-(4-Chlorophenyl)cyclohexyl]-3-~(2-methylaminoethoxy)carbon-
yloxy]-1,4-naph~hoq~inone hydrochloride, m.p. 2Ll-213 C, NMR ~H (d6
DMSO) 7.95-8.05 (2H, m), 7.74-7.9 (2H, m), 7.33 (4H, s), 4.2-4.3 (2H,
m), 3.5-3.6 (2H, m), 3.0-3.2 (lH, m), 2.75 (3~, s), 2.5-2.7 (lH, m),
2.05-2.3 (2H, m), 1.8-2.0 (2H, m), 1.4-1.7 (4H, m).
2-i~rans-4-~4-Chlorophenyl)cyclohexyl]-3-[N.N.-bis (2-aminoethyl) car-
bamoyloxyj-1,4-naphthoquinone dihydrochloride dihydrate, m.p.
~~ -215-218 C, ~MR ~H (d6-DMS0)8.2S-8.7 (4H, br) 7.85-8.15 (4H, m),
7.3-7.45 (4H,m) 3.5-4.0(4H,m), 3.0-3.4 (5H, m) 2.7 (lH, m), 1.5-2.2
(8H, m)
,
- ~ Exam~le 3
- ~~ ~ 2 - ~ trans - t ~ - tert - Butv l ~Yc loh~xy~ e thvl l - 3 - ~ N - me thyl - N - ~ 2 - ~ N
,-methvl-N--~ert-~e~arbonylamino~ethvllcarbamovloxy!-l.4- ,
na~hthoauinone
',' ~ ?'~
~'0 93t20044 PCT/GB93/00708
o a stirred solution or ~-itrans-(~-ter~-butylcvclohexyl~ethyl]-
~-nvdroxy-i,4-naph~hoquinone (6.76:19mM) in drv dichloromethane (2S0
mL) under nitrogen, was added phosgene (20 mL; 12.5% solution in
~ol~ene), _ollowed bv pyridine (1.6 ml.20 mM) added dropwise, and the
mixture was stirred for 1 hour. A solution of pyridine (1.6 mL; 20~M)
and tert-butyl-N-me~hyl-N-[2-(~ethylamino)ethyl]carbamate (3.7g;21mM)
was added dropwise. and ehe reaction ~ixture stirred at room
temperature for a further 3.S hours. Evapora~ion of the solvents in
vacuo gave a yellow solid, which was chromatographed on silica.
elut.r.g with toluene and ~hen 1:8 ethyl acetate:toluene, to give ,he
.itle compound (3.19g; 31~) m.p. 117-118 C, NMR ~H (CDCl3) 8.05-8.15
(2H,mj, 7.65-7.75 (2H,m), 3.45-3.6 (4H, m), 3.2-3.1 (3H, ~xs),
2.95(3H, 2xs), 2.45-2.;5 (2H, m), 1.7-l.9 (5H, m) 1.5 (9H, 2xs),
0.9-1.1(5H, m), 0.85(9H,s).
'.:
Exam~le 4
2-~trans-(4-tert-Butvlcyclohex~l)methvll-3-(N-methYl-N-~2-tmethYla-
ino)ethvll carbamovloxY)-l~ na~hthoquinone hvdrochloride ~
`
2-f~3L~-(4-tert-butylcyclohexyl)methyl]-3-(N.methyl-N-[2- ;j;
(me~hylamino)ethyl] carbamoyloxy)-l, 4-naphthoquinone (lg) was added
,o ether satura~ed with HC1(20 ~1) and the solution scirred at room
temperature in the dark for 1.5 hours. The ether was evaporated ~
acuo to leave a yellow oil, which was dissolved in ethyl acetate and
filtered. Cyclohexane was atded to the filtrate, resulting in the
precipitation of thé eitle co~pound as a yellow powder (0.4ig) m.p.>
140 C (decomposes).
,
..
