Note: Descriptions are shown in the official language in which they were submitted.
LZ~i2~P
HETERO POLYCYCLIC BIOCIDAL COM~OUNDS. THEIR SYNTHESIS
AND INTE~MEDIAI'ES THEREFORE. FORMULATIONS CONTAINING THEM
AND_THEIR USE IN MEDICINE
The present invention relates to heteropolycyclic aromatic alkanol
derivatives which have been found to have biocidal activity. More
specifically the invention concerns aminoalkanol derivatives
containing a heteropolycyclic aromatic ring system, methods for the
synthesis thereof, novel intermediates thereof, pharmaceutical
formulations thereof and the use thereof as biocidal agents,
particularly antitumor agents.
We have discovered a novel class of heteropolycyclic aromatic alkanol
derivatives which have biocidal activity. Accordingly, in a first
aspect, the present invention provides a compound of the formula (I)
ArCH2NHR (I)
or a monomethyl or monoethyl ether thereof, the compound of
formula (I) including these ethers containing no more than 29 carbon
atoms in totàl, or an ester or a salt thereof;
wherein Ar is a fused tetracyclic hetero aromatic ring system of the
formula:
~r~ CY
in which Z is oxygen, sulphur or a group NRl wherein Rl is hydrogen,
methyl or ethyl and ) CY is a bicyclic aromatic ring system
AJR/JJ/6th March, 1990.
PB1125
~L2~i2~
comprising a phenyl ring and a 5-membered ring system which contains
one heteroatom ~1 selected from oxygen, sulphur or a group NR2 wherein
R is hydrogen, methyl or ethyl; the tetracyclic ring system being
optionally substituted by one or two substituents; said substituents
containing not more than four carbon atoms in total when taken
together and are the same or different each being selected from
halogen; cyano; Cl 4 alkyl or Cl 4 alkoxy, each optionally substituted
by hydroxy or Cl 2 alkoxy; halogen substituted Cl 2 alkyl or Cl 2
alkoxy; a group S(O) R wherein n is an integer 0,1 or 2 and R i5
Cl 2 alkyl optionally substituted by hydroxy or Cl 2 alkoxy; or Ar is
optionally substituted by a group NR R containing not more than 5
carbon atoms wherein R and R are the same or different and each is a
Cl 3 alkyl group or ~R R5 forms a five-or six-membered heterocyclic
ring optionally containing one or two additional heteroatoms;
R contains not more than eight carbon atoms and is a group
R10
R6 I R12
(I12)m or - C ~ R13
R - C - R OH
I
OH
wherein m is 0 or l;
R6 is hydrogen or Cl 3 alkyl optionally substituted by hydroxy;
R and R are the same or different and each is hydrogen or Cl 3
alkyl;
R9 is hydrogen, methyl or hydroxymethyl;
-C-C- is a five-or six-membered saturated carbocyclic ring;
R10, Rll and R12 are the same or different and each is hydrogen or
methyl;
AJR/JJ/6th March, 1990.
PB1125
2~ 6
R13 is hydrogen, methyl, hydroxy, or hydroxymethyl; R14 is hydrogen,
methyl, hydroxy or hydroxymethyl.
Suitably the tetracyclic aromatic ring system Ar is unsubstituted or
has only one substituent. Preferably the aromatic ring is
unsubstituted.
Suitably ArCH2~HR or a monomethyl or monoethyl ethPr -thereof contains
not more than 28 carbon atoms in totai.
Suitably m is 0.
Suitably R is
- C - R16 or
H - C - R OH
OH
wherein R is CH2H' CH(CH3)0H or CH2CH2H'
R is hydrogen, Cl 3 alkyl or CH20H,
R is hydrogen or methyl.
Suitably R is CH20H or CH(CH3)0H. Suitably R is hydrogen,
methyl, ethyl or CH20H.
Preferably R is
CH OH
~ 2 18
H - C - R17
1H
wherein R17 is hydrogen or methyl and Rl is hydrogen, methyl or
ethyl, preferably methyl.
AJR/JJ/6th March, 1990.
PB1125
- 4 -
Tetracyclic heteroaromatic ring systems of compounds of the present
invention have ~he following nomenclature:
Z = S, Z = NMe,
10-methyl-lOH-[l]benzothieno[3,2 -k]
indol-6-yl
8 ~ Z ~ ~ 2 Z = S, zl = NNe,
10-Methyl;lOH-[ljbenzothieno[3,2-k]
z = O, zl = NMe,
2-(lo-methyl-loH-benzofurc-(3~2 -k)
indol-6-yl
Z- O, Z =0,
2-(benzofuro(5,6-k)benzofuran-4-yl,
~ 10
~/ \ ~ ~ zl~ r Z ~ O, zl c O
1 ~ z J ~ 1 2-(benzofuro(5,6-b)benzofuran-8-yl-,
6 5 4 z _ O, zl _ O
2-(benzofuro(5,6-_)benzofuran-2-yl-,
: 2 z = O, zl = O,
1 ~ 2-(benzofuro(5,4-k)benzofuran-2-yl-,
l 3
~\~ Z, o~ Zl = o
8 ~ ~ 4 2-(benzofuro(5,4-k)benzofuran-9-yl-,
7 6 5
Z= O, Z - O,
2-(benzofuro(5,4 k>benzofuran-4-yl,
Z = o~ Zl = o~
2-(benzofuro(5,4-k)benzofuran-8-yl-,
~JR/JJ/6th March, 1990.
,:
.- . , , ~
.
..
:
PB1125
- 5 20~26%6
Z ~ S, Z ~ NMe,
l-Methyl-lH-[l]benzothieno[2,3-g]
indol-3-yl-
_1~ z e o ~ zl e NMe,
g ~ / ~ l Methyl-lH-benzofuro[2,3-g]indol-
7 6 5 z e NEt, Z - NMe,
6-Ethyl 1,6-dihydro-'1-methylpyrrolo
[3,2-c]carbazol-3-yl-
z e S ~ Z e NMe,
3-Methyl-3H-[l]benzothieno[2 7 3-e]
indol-l-yl-
7 ~ ~ 4 Z - ~Me, Z ~ S
¦ O ¦ O 10-Methyl-lOH-thieno[3,2-a]carbazol-
8 ~ ~` Z ~ ~ 2-yl-
1 Z - NMe, Z - S,
10-Methyl-lOH-thieno[3,2-a]carbazol-
4-yl-
Speciflc compounds within the scope of formula (I) include:
2-(((10-Methyl-lOH-[l]benzothieno[3,2-b]indol-3-yl)methyl~-amino)-2-
methyl-1,3-propanediol,
2-(((10-Methyl-lOH[l]benzothieno[3,2-k]indol-6-yl)methyl)amino)-2-
methyl-1,3-propanediol,
2-Methyl-2-(((10-methyl-lOH-benzofuro(3,2-k)indol-6-yl)methyl)amino)-
1,3-propanediol,
AJR/JJ~6~h March, 1990.
PB1125
, - 6 -
2-(((Benzofuro(5,6-b)benzofuran-4-yl)methylamino)-2-methyl-1,3-propan-
ediol,
2-(((Benzofuro(5,6-_)benzofuran-8-yl)methyl)amino)-2-methyl-1,3-
propanediol,
2-(((Benzofuro(5,6-b)benzofuran-2-yl)methyl)amino~-2-methyl-1,3-propa-
nediol,
2-(((~enzofuro(5,4-b)benzofuran-4-yl)methyl)amino)-2-methyl-1,3-propa-
nediol,
2-((~Benzofuro(5,4-b)benzofuran-9-yl)methyl)amino)-2-methyl-1,3-propa-
nediol,
2-(((Benzofuro(5,4-b)benzofuran-8-yl)methyl)amino)-2-methyl-1,3-propa-
nediol,
2-(((Benzofuro(5,4-_)benzofuran-2-yl)methyl)amino-2-methyl-1,3-propan-
ediol, and monomethyl or monethyl ethers, esters, and addition salts
thereof.
Salts included within the scope of the present invention are those of
compounds of formula (I) and ethers and esters thereof.
Esters and non-pharmaceutically useful salts of the compo~mds of the
foxmula (I) are useful intermediates in the preparation and
purification of compounds of the formula (I) and pharmaceutically
useful salts thereof, and are therefore within the scope of the
present inventlon. Thus, salts of the compounds of the formula (I)
useful in the present invention include but are not limited to those
derived from inorganic acids, such as hydrochloric, hydrobromic,
sulfuric and phosphoric acids, and organic acids such as isethionic
(2-hydroxyethylsulfonic), maleic, malonic, succinic, salicylic,
tartaric, lactic, citric, formic, lactobionic, pantothenic,
methanesulfonic, ethanesulfonic, benzenesulfonic, ~-toluenesulfonic,
naphthalene-2-sulfonic, and ascorbic acids, and amino acids such as
glycine.
Pharmacologically and pharmaceutically acceptable salts are preferred,
particularly those that are soluble in solvents suitable for
parenteral administration, for example, hydrochlorides, methanesul-
AJR/JJ/6th March, 1990.
PB1125
- 7
fonates and isethionates.
Esters of compounds of formula (I) are derived from acids known to
those skilled in the art to be suitable for ester formation, and are
conveniently those derived from Cl 6 alkanoic acids or alkanoic acid
derivatives, for example acetic acid, propionic acid, _-butyric acid
and iso-butyric acid. The esters may be formed from all or only some
of the hydroxy groups contained in the compounds of formula (I).
The compounds of formula (I) and their ethers, esters, and salts
thereof may be prepared by any method known in the art for the
preparation o compounds of analogous structure. Thus, the compounds
of formula (I) may, for example, be prepared by any of the methods
defined below.
l. The reduction of a compound Ar~CH-NR (II)
wherein Ar and R are as hereinbefore defined or an appropriately
protected derivative thereof followed by deprotection where
appropriate.
The conditions and reagents for such a reaction are well known to
those skilled in the art, and any such conditions/reagents may be
employed tha~ will not reduce the aromatic ring system. The
conversion of (II) or suitably protected derivatives thereof may be
carried out by a reducing agent followed by deprotection if necessary
The reduction is conveniently carried out by a m~tal hydride such as
lithium aluminum hydride, sodium borohydride, sodium cyanoborohydride,
or by catalytic hydrogenation, conveniently by hydrogen in the
presence of a metal catalyst such as palladium or platinum, or
equivalent reagents as outlined by J. March, Advanced Or~anic
hemistrv, 2nd ed., pages 819-820, McGraw Hill, New York, 1977. The
reduction is suitably carried out with Ar-CH=NR in solution in an
inert solvent or mixture of solvents compatible with the reducing
AJR/JJ/6th March, 1990.
PE1125
Z1~2~
agent, at a non-extreme temperature, or example, between 0 and 80C,
conveniently at room temperature.
In the case of lithium aluminum hydride and like reagents, suitable
solvents include ethers (for example tetrahydrofuran, diethyl ether
and 1,2-dimethoxyethane~ optionally in the presence of a hydrocarbon
cosolvent (for example toluene, benzene or hexane).
In the case of sodium borohydride and like reagents, suitable solvents
include alcohols (for example ethanol, methanol or isopropanol)
optionally in the presence of a hydrocarbon cosolvent (for example
toluene, benzene or hexane) or an ether cosolvent (for example
diethylether or tetrahydrofuran).
In the case of sodium cyanoborohydride and like reagents, suitable
solvents include those described for sodium borohydride and the
reduction is conveniently carried out in the presence of an acid,
conveniently glacial acetic acid or ethanollc hydrochloric acid as
outlined in, for example, R. Hutchins et al., Organic Preparations and
Procedures International_ll, 201 (1979).
In the case of catalytic hydrogenation, sultable solvents include
alcohols (for example methanol and ethanol) optionally in the presence
of a hydrocarbon cosolvent (for example toluene or benzene), or ether
cosolvent (for example diethyl ether or tetrahydrofuran) optionally in
the presence of an acid (for example glacial acetic acid or ethanolic
hydrochloric acid), or glacial acetic acid;
Protected derivatives of compounds ArCH=NR are conveniently used when
lithium aluminum hydride is employed as the reducing agent.
Convenient protecting groups are compatible with the reducing agent
utili~ed and are readily removed under nondestructive conditions, for
example ben~yl, tetrahydropyranyl and isopropylidene ethers.
AJR/JJ/6th March, 1990.
PB1125
g
62~
It is often convenient not to isolate the compound ArCH~NR but to
react a compound ArCHO with a compound NH2R; wherein Ar and R are as
defined in (I) and to reduce the compound ArCH=NR so formed i situ.
The reaction of the compounds ArCHO and NH2R is again suitably carried
out using conditions and reagents which are well known to those
skilled in the art, for example in the presence of an acid, such as a
sulfonic acid, i.e. ~-toluenesulfonic acid, in an appropriate inert
solvent, such as an aromatic hydrocarbon, suitably toluene, with
azeotropic removal of water followed by treatment with the reducing
agent in an approprlate solvent, suitably ethanol or ~ethanol.
Alternatively, ArCH~NR formed under equilibrium conditions in
appropriate solvents can be reduced in situ with an appropriate
reducing agent, suita~ly sodium cyanoborohydride.
The compound ArCHO may be in the form of a protected aldehyde, for
example an acetal, which liberates the aldehyde functionality under
the reaction conditions. In turn, ArCHO can be synthesized by reacting
the appropriate aromatic polyheterocycle optionally substituted with a
carbalkoxy group with a formylatlng agent such as that generated by
the react~on between SnC14 and C12CHOCH3 or equivalent reagents,
for e~ample, according to the method of A. Reiche et al., Chem. Ber.
93, 88 (1960), or with other standard formylating reagents/procedures
known ~o the art, for exa~nple, the Gatterman-Koch reaction
(CO/HCl/AlC13/CuCl), the Gatterman reaction (HCN/HCl/ZnC12), and the
Vilsmeier reaction (POC13/Ph~(Me)CHO or POC13/Me2NCHO) (J. ~arch,
vide s~pr_ pages 494-497).
The compounds ArCHO may also be prepared from an appropriate aromatic
heteropolycycle substituted by a suitable functional group and
converting this functional group to an aldehyde group by methods well
known to those skilled in the art. Suitable functional groups include
CHBr2, CH3, CoR19 wherein Rl is a primary or secondary Cl 6 alkyl
group, COOH or a derivative thereof such as an ester, amide or acid
chloride or CN.
AJR/JJ/6th March, l99O.
PB1125
- 10 - ~ 2~;
Where the aromatic heteropolycycle bears substituents, ArCHO may be
prepared by a variety of methods known in the art of organic chemistry
depending on the nature of the substituent on the ring. For example,
if the substituent(s) is a halogen, the starting m~terials may be
prepared by direct treatment oE the aromatic heteropolycycle ring with
a halogenating agent (e.g. Cl2, Br2, or S02C12) or indirectly by such
routes as the Sandmeyer reaction (H.H. Hodgson, Chem. Rev. 40, 251
(1947). If the substituent(s) is alkyl, the aromatic heteropolycycle
may be reacted with the appropriate reagents under Friedel-Crafts
reaction conditions (G.A. Olah, Friedel Crafts and Related Reactions,
Vols. 1-3, Interscience, New York, NY, 1963-1965).
The compounds NH2R also may be prepared by methods known in the art,
for example when R is as hereinbefore defined by the reaction of
N02R wherein R is a group
R6
- CH - R
( CH2 ) m
OH
and R6 to R9 and m are as hereinbefore defined with an appropriate
aldehyde, conveniently acetaldehyde or formaldehyde (as in
B.M. Vanderbilt and H.B. Hass, Ind._~g. Chem. 32, 34 (1940)) followed
by reduction (as outlined in J. March, vide supra, pages 1125-1126),
conveniently by hydrogen and a metal catalyst (for example, a platinum
containing catalyst) in an appropriate solvent, conveniently glacial
acetlc acld.
2. The reduction of a compound Ar.CO.NHR; wherein Ar and R are as
hereinbefore defined and the hydroxy groups are optionally
protected, followed by deprotection of the hydroxy groups where
AJR/JJ/6th March, 1990.
PB1125
- 11 -
L26~Ç;
appropriate. The reduction may be carried out by standard
reducing agents known for carrying out this type of reduction
that will not reduce the aromatic ring system (as outlined in J.
March, vide supra, page 1122), for example, a hydride reagent
such as lithium aluminium hydride in an inert solvent, such as an
ether, i.e. tetrahydrofuran, at a non-extreme te~perature, Eor
example, at between 0 and 100 C and conveniently at the reflux
temperature of the ether.
The compound Ar.CO.NHR may be formed by the reac~ion of the
appropriate acid (ArCOOH) or a suitable reactive acid derivative
thereof as outlined in J. March, vide supra, pages 382-390) for
example, an acid halide in an inert solvent, with an amine NH2Rl
in which the hydroxy groups are optionally protected, for
example, when the compound NH2Rl is a diol, by an isopropylidene
group. The compound Ar. CO. NHR so formed is suitably reduced
in ~ and deprotected if necessary to give a compound of
formula (I). The compounds of the formula ArCOOH can be prepared
by methods well known to those skllled in the art.
3. The reaction of a compound ArCH2L (wherein Ar is as hereinbefore
defined and L is a leaving group), with a compound NH2R as
hereinbefore defined. Suitable leaving groups are those defined
by J. March, vide ~ pages 325-331, and include halogens such
as chlorine or bromine and sulfonic acid derivatives such as
D-toluenesulfonate. The reaction is suitably carried out in an
appropriate solvent, such as a dipolar aprotic solvent or alcohol
at a non-extreme temperature, for example 50-100. The compounds
of the formula ArCH2L can be prepared by methods well known to
those skilled in the art.
There is therefore provided,` as a further aspect of the invention, amethod for the preparation of a compound of formula (I) comprising any
method known for the preparation of analogous compounds, in partlcular
those methods defined in (1) to (3) hereinabove.
AJR/JJ/6th March, 1990.
PB1125
- 12 -
In a further aspect, the present invention provides novel chemical
intermediates of the formulae Ar.CH=NR, Ar.CO.NHR or ArCH2L as herein
before defined.
Compounds of the formula (X) in which one or more hydroxy groups are
protected, for example by benzyl or trityl groups or by an
isoproplylidene group are also useiul intermediates in the preparation
of compounds of the present invention.
The compounds of this invention have biocidal activity, e.g. are toxic
to certain living cells which are detrimental to mammals, for example
pathcgenic organisms and tumours.
This toxicity to pathogenic organisms has been demonstrated, for
example, by activity against one or more of the following: viruses
(e.g. Herpes ~oel~ vero), fungi (e.g. Candida albicans), protozoa
(e.g. Eimeria tenella and Trichomonas .va~inalis), bacteria (e.g.
Mycoplasma smegmatis and Streptococcus pyog~), and helminths (e.g.
NipRostrongvlus brasiliensis and Bru~La pahan~i). The antitumour
activity of compounds of formula I has been demonstrated in a number
of recognlzed screens and primarily by activity against ascitic P388/0
leukaemia.
Preferred compounds of the formula (I) are those which have antitumour
activity. The activity against ascitic tumours, including P388/0, is
evidenced by reduction of tumour cell number in mammals (for example,
mice bearing ascitic tumou~s) and their consequent increase in
survival duration as compared to an untreated tumour bearing control
group. Antitumour activity is further evidenced by measurable
reduction in the size of solid tumours following treatment of mammals
with the compounds of this invention compared to the tumours of
untreated control tumour bearing animals. Compounds of formula (I)
are active against murine tumours such as lymphocytic leukaemia
P388/0, lymphocytic leukaemia L1210, melanotic melanoma B16, P815
mastocytoma, MDAY/D2 fibrosarcoma, colon 38 adenocarcinoma, M5076
rhabdomyosarcoma and Lewis lung carcinoma.
AJ~/JJ/6th March, 1990.
PB1125
- 13 -
26~
Activity in one or more of these tumour tests has been reported to be
indicative of antitumour activity in man (A. Goldin et al. in Methods
in Cancer Research ed. V.T. DeVita Jr. and H. Busch, 16, 165, Academic
Press, N.Y. 1979).
There are subline~ o~ P388/0 which have been made resistant to the
following clinically useEul agents: cytosine arabinoside,
doxorubicin, cyclophosphamide, 1 phenylalanine mustard, methotrexate,
5-fluorouracil, actinomycin D, cis-platin and bis-chloroethylnitro-
sourea. Compounds of this invention show potent activity against
these drug-resistant tumours using the procedure for P388/0 above.
Compounds of formula (I) have also been Eound to be active against
human tumour cells in primary cultures of lung, ovary, breast, renal,
melanoma, unknown primary, gastric, pancreatic, mesothelioma, myeloma,
and/or colon cancer. (As used herein "cancer" is to be taken as
synonymous with "malignant tumour" or more generally "tumour" unless
otherwise noted.) This is a procedure in which the prevention of
tumour cell colony formation, i.a. tumour cell replication, by a drug
has been shown tc correlate with clinical antitumour activity in man
(D.D. Von Hoff et al., Cancer Chemotheray~ and Pharmacologv 6, 265
(1980); S. Salmon and D.D. Von Hoff, Seminars in Oncolo~v, 8, 377
(1981)).
Compounds of formula I which have been found to have antitumour
ac~ivity intercala~e in vitro with DNA (this property is determined by
viscometric methods using the procedure of U. D. Wilson et _1.,
~ucleic Acids Research 4, 2697 (1954)) and have a log~P as calculated
by the method of C. Hansch and A. Leo in Substituent Constants for
Correlation AnalYsis in Chemistry_ and Biology, John Wiley and Sons,
New York, 1979, lying in the range between -2.0 and +2.5.
As has been described above, the compounds of the present invention
are useful for the treatment of tumours. The invention thus further
provides a method for the treatment of tumours in animals, including
AJR/JJ/6th March, 1990.
PB1125
- 14 -
2~ 6~Ç~
mammals, especially humans, whi.ch compr~ses the administration of a
clinically useful amount of compoun of formula (I) in a pharmaceutic-
ally useful form, once or several times a day or other appropriate
schedule, orally, rectally, parenterally, or applied topically.
In addition, there is provided as a further, or alternative, aspect of
the invention, a compound of formula (I) for use in therapy, for
example as an antitumour agent.
The amount of compound of formula (I) required to be effective as a
biocidal agent will, of course, vary and is ultimately at the
discretion of the medical or veterinary prac~itioner. The factors to
be considered include the condition being treated, the route of
administration, and nature of the formulation, the mammal's body
weight, surface area, age and general condition, and the particular
compound to be administered. A suitable effective antitumour dose is
in the range of about 0.1 to about 120 mg/kg body weight, preferably
in the range of about l.S to S0 mg/kg, for example 10 to 30 mg/kg.
The total daily dose may be given as a single dose, multiple doses,
e ~,, two to six times per day or by intravenous infusion for selected
duration. For example, for a 75 kg mammal, the dose range would be
about 8 to 9000 mg per day, and a typical dose would be about 2000 mg
per day. If discrete multiple doses are indicated, treatment might
typically be 500 mg of a compound of formula I given 4 times per day
in a pharmaceutically useful formulation.
Whilst it is possible for the active compound (defined herein as
compound of formula (I), or ether, ester, or salt thereof) to be
administered alone, it is preferable to present the active compound in
a pharmaceutical formulation. Formulations of the present invention,
for medical use, comprise an active compound together with one or more
pharmaceutically acceptable carriers thereof and optionally other
therapeutical ingredients. The carrier(s) must be pharmaceutically
acceptable in the sense of being compatible with the other ingredients
of the formulatLon and not deleterious to the recipient thereof.
AJR/JJ/6th March, 1990.
PB1125
- 15 -
62~i
The present invention, therefore, further provides a pharmaceutical
formulation comprising a compound of formula (I) (in the form of the
free base, ether, or ester derivative or a pharmaceutically acceptable
acid addition salt thereof) together with a pharmaceutically
acceptable carrier therefor.
There is also provided a method for the preparation of a
pharmaceutical formulation com2rising bringing into association a
compound of formula (I) an ether, ester, or pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable carrier
therefor.
Whilst the antitumour activity of the compounds of Eormula (I) is
believed to reside in the free base, it is often convenient to
administer an acid addition salt of a compound of formula (I).
The formulations include those suitable for oral, rectal or parenteral
(including subcutaneous, intramuscular and intravenous)
administration. Preferred formulations are those suitable for oral
or parenteral administration.
The formulations may conveniently be presented in unit dosage form and
may be prepared by any of the methods well known in the art of
pharmacy. All methods lnclude the step of b~inging the active
compound into association with a carrier which constitutes one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing the active compound into assocation
with a liquid carrier or a inely divided solid carrier or both and
then, if necessary, shaping the product into desired formulations.
Formulations of the present invention suitable for oral administration
may be presented as discrete units such as capsules, cachets, tablets
or lozenges, each containing a predetermined amount of the active
compound; as a powder or granuies; or a suspension in an aqueous
AJR/JJ/6th March, 1990.
PB1125
- 16 - ~ ~ ~Z~6
liquid or non-aqueous liquid such as a syrup, an elixir, an emulsion
or a draught.
A tablet may be made by compression or moulding, optionally with one
or more accessory ingredients. Compressed tablets may be prepared by
compressing in a suitable machine the active compound in a free-
flowing form such as a powder or granules, optionally mixed with a
binder, lubricant, inert diluent, surface active or dispersing agent.
Moulded tablets may be made by moulding in a suitable machine, a
mixture of the powdered active compound with any suitable carrier.
A syrup may be made by adding the active compound to a concentrated,
aqueous solution of a sugar, for example sucrose, to which may also be
added any accessory ingredients. Such accessory ingredient(s) may
include flavourings, an agent to retard crystallization of the sugar
or an agent to increase the solubility of any other ingredient, such
as a polyhydric alcohol for example glycerol or sorbitol.
Formulations for recta]. administration may be presented as asuppository with a conventional carrier such as cocoa butter.
Formulations suitable for parenteral administration conveniently
comprise a sterile aqueous preparation of the active compound which is
preferably isotonic with the blood of the recipient. Such
formulations suitably comprise a s~lution of a pharmaceutically and
pharmacologically acceptable acid addition salt of a compound of the
formula (I) that is isotonic with the blood of the recipient. Thus,
such iormulations may conveniently contain distilled water, 5~
~extrose in distilled water or saline and a pharmaceutically and
pharmacologically acceptable acid addition salt of a compound of the
formula (I) that has an appropriate solubility in these solvents, for
example the hydrochloride, isethionate and methanesulfonate salts,
preferably the latter.
~JR/JJ/6th March, 1990.
PB1125
- 17 -
2C~ i2ti
Useful formulations also comprise concentrated solutions or solids
containing the compound of formula (I) which upon dilutlon with an
appropriate solvent give a solutlon suitable for parenteral
ad~inistration as above.
In addition to the aforementioned ingredients, the fonnulations of
this invention may further include one or more accessory ingredient(s)
selected from diluents, buffers, fla~ouring agents, binders, surface
active agents, thickeners, lubricants, preservatives (including
antioxidants) and the like.
The following examples are provided by the way of illustration of the
present invention and should in no way be construed as a limitation
thereof.
General Comments
All solvents were reagent grade and used without further purification
with the following exceptions. Tetrahydrofuran (THF) was dried by
distillation from Na/K alloy under nitrogen (N2) and used immediately.
Toluene (PhCH3) was distilled from CaH2 under N2 and stored over 3A
molecular sleves. Chemicals used were reagent grade and used without
purification unless noted. The full na~e and address of the suppliers
of the reagents and chemicals is given when first cited. After this,
an abbre~iated name is used.
Preparative HPLC was carried out on a Waters Prep LC~System 500A
machine using two 500 g silica gel (SiO2) cartridges unless otherwise
noted. Plugs of SiO2 used for purifications were "flash
chromatography" silica gel (Merck Co., Inc., ~erck Chemical Division,
Rahway, NJ, 07065 silica gel 60, 230-400 mesh). An appropriate volume
sintered glass funnel was filled approximately 3/4 full with the SiO2
and packed evenly by tapping the outside of the funnel. A piece of
filter paper was then placed on top of the SiO2 and a solution of the
material to be purified applied evenly to the top. Gentle suction
AJR/JJ/6th March, 1990.
- 18 - PB1125
2~3L2~2~
through a filter flask moved the eluting solvent through the plug
rapidly. The appropriate size fractions were combined as needed and
further manipulated.
General proceduxes are dascribed in detail. Analogous procedures show
melting point ~mp), recrystallization solvents, and elemental analyses
(all elements analyzing within a difference of ~ 0.4~ of the expec~ed
value). Any changes to the procedure such as solvent, reaction
temperature, reaction time, or workup are noted.
1~
NMR ( H, C), IR, MS data of all new products were consistent with
the expected and proposed structures. The positions assigned to
structural isomers were unequivocally determined by a number of NMR
techniques. All final products were dried in a vacuum oven at 20 mm
Hg pressure at the temperature indicated overnight (12-16 h). All
temperatures are in degrees Celsius.
ExamEle 1
2-Meth~1-2-(((10-methy~-lOH-benzofuro(3.2-b)indol-6-vl~methyl)amino)-
1.3-propanediol
lA 2-(2-Nitrophenyl)benzofuran
To a RB flask equipped with a magnetic stirring bar, rubber
septum and N2 inlet line was added a solution of benzofuran
(~ldrich Chemical Co., P.O. Box 2060, Milwaukee, WI, 53201, 34.26
g, 0.29 mol) in Et20 (800 ml). A solution of _-BuLi (Aldrich,
20.5 g, 0.32 mol) in hexane (200 ml) was added to the flask via
cannula. The mixture was stirred for 30 min. at RT. The mixture
was then added via cannula to a RB flask equipped with a magnetic
stirring bar, rubber septum and N2 inlet line that contained a
solution of 2-fluoronitrobenzene ~Aldrich, 37.26 g, 0.264 mol) in
Et20 (800 ml) cooled to -40 (CH3CN/dry ice bath). The resulting
mixture was istirred overnight at RT. H20 (40 ml) was then added
AJR/JJ/6th Narch, 1990.
. .
PB1125
- 19 -
to the flask to destroy excess _-BuLi. The mixture was then
further diluted with H2O (1 L), the two layers separated and the
H20 layer extractad with Et20 ~2x500 ml). The organic layers
were combined, dried (Na2SO4) and concentrated to give a dark red
oil. This material was passed through a plug of SiO2 using
hexane/EtOAc (9:1) as the eluting solvent. The appropriate
fractions were combined and the solvent removed to give 34.0 g
(49.7~ yield) of crude sclid that was used in the next step
without additional puriflcation. A portion was washed with
hexane to give 2-(2-nitrophenyl)benzofuran, mp 79-80.5 (C,H,N).
lB 10H-Benzofuro(3,2-b)indole
To a RB flask equipped with magnetic stirring bar, reflux
condenser and N2 inlet line was added 2-(2-nitrophenyl)benzofuran
(lA,70.0 g,O.29 mol) and triethylphosphite (Aldrich, 98.0 g,
0.59 mol). The resulting mixture was heated at 100-110 for 7 h.
The mixture was then dissolved ~n EtOAc (500 ml) and passed
through a plug oE SiO2 using EtOAc as the eluting solvent. The
appropriate fractions were combined and concentrated to give a
crude material which was dissolved :Ln a mixture of hexane/EtOAc
(1:1) and passed through a plug of S:L02 using hexane/EtOAc (9:1)
as the eluting solvent. The appropr:Late fractions were combined
and concentrated to give 53.5 g ~89~ yield) of crude solid that
was used in the next step without additional purification. A
portion was recrystallized from EtOAc/hexane (1:6) to give
lOH-benzofuro(3,2-b)indole, mp 197.5-198 (C,H,N).
lC 10-Methyl-lOH-benzofuro~3,2 b)indole
To a RB flask equipped with magnetic stirring bar, reflux
condenser, rubber septum and N2 inlet line was added a 60% oil
dispersion of NaH (Aldrich,5.78 g,0.151 mol). The oil was
removed from the dispersion by washing with hexane (3x100 ml).
Dry THF (200 ml) was then added to the flask via cannula to cover
AJR/JJ/6th March, 1990.
,,
,
:
PB1125
- 20 -
2~2~6
the NaH. A solution of lOH-benzofuro(3,2-b)indole ~lB,28.4
g,O.137 mol) in THF (200 ml) was added to the flask via cannula.
The mixture was stirred for 15 min. at RT. Dimethyl sulfate
(Aldrich,l9.0 ~, 0.151 mol~ was then added to the flask by
syringe and the resulting mixture was stirred 90 min. at RT.
H20 (10 ml) was added to the flask to destroy excess NaH. The
mixture was diluted with 0.1 N HCl ~1 L) and extracted with EtOAc
(1 L). The organic layer was washed with H20 (2xl L), dried
(Na2S04) and concentrated to give 35.6 g of crude material. This
material was passed through a plug of SiO2 using PhCH3 as the
eluting solvent. The appropriate fractions were combined and the
sol~ent removed to give after drying 29.0 g (96.0~ yield) of
10-methyl-10_-benzofuro-(3,2-b)indole, mp 114-115 (C,H,N).
lD 10-Methv:L-lOH-benzofuro(3.2-b)indole-6-carbaldehvde
To a RB flask equipped with a magnetic stirring bar, rubber
septum, and N2 inlet line was added a solution of 10-methyl-
lOH-benzofuro(3,2-b)indole (lC,10.0 g,44 mmol) in dry THF (1 L).
The mixture was cooled to -78, then a solution of s-BuLi
(Aldrich,1.3 M in cyclohexane, 8.~ mmol,6.8 ml) was added by
syringe. The resulting mixture was allowed to warm to -30 then
recooled to -78 before adding D~F (Aldrich,0.67 g,9.2 mmol) by
syri~ge. The resulting mixture was stirred overnight at RT. H20
(10 ml) was added to the flask to destroy excess s-BuLi. The
mixture was concentrated and the residu~ was partitioned between
EtOAc (1 L) and 0.2 N HCl (1 L). The organic layer was washed
with satd. NaCl solution, dried, (Na2S04), and concentrated to
give a yellow solid which was triturated with Et20 ~300 ml) to
yield after drying 9.4 g (~6.0%) of 10-methyl-lOH-benzofuro-
(3,2-b)indole-6 carbaldehyde, mp 114.5-115.5 (C,H,N).
lE 2-Methyl-2~ 10 methyl-lOH-benzofuro~3~-b~indol-6-yl~meth~yl)
amino~-1.3-propanediol Hvdrochloride
AJR/JJ~6th March, 1990.
~ 2~ PBl125
- 21 -
To a 3-necked RB flask equipped with a magnetic stirring bar,
condenser, thermorneter, Dean-Stark trap and N2 inlet line was
added 10-methyl-lOH-benzofuro(3,2-b)indole-6-carbaldehyde (lD,
9.3 g,37.3 mmol), 2-amino-2-methyl-1,3-propanediol (Aldrich,7.~4
g,74.6 mmol), p-toluenesulfonic acid monohydrate (Aldrich,0.1 g)
and PhCH3 (300 ml). The mixture was stirred at reflux with
azeotropic removal of H2O for 2.5 h. Most of the PhCH3 was then
removed by distillation. The mixture was then cooled in a~ ice
bath and diluted with abs. EtOH (300 ml) and further cooled.
Solid NaBH4 (Aldrich,2.82 g,74.6 mmol) was added in one portion
to the reaction mixture. The ice bath was then removed, the
reaction mixture allowed to warm to RT and stirred overnight.
The crude reaction mixture was concentrated to dryness, vigor-
ously stirred with H2O (500 ml) and filtered. After thoroughwashing with H2O and drying, the resulting solid crude free base
was converted to its HCl salt by treatment with a 5N solution of
g. HCl in absolute EtOH. The solution was filtered through a
medium porosity sintered glass funnel and precipitated by
addition of Et2O (final volume 2 L). Recrystallization twice
(CH3OH/Et2O) followed by filtration and drying gave 5.7 g (40.8
yield) of 2-methyl-2-(((10-methyl-10,H-benzofuro(3,2-b)indol-6-yl)
methyl)amino)-1,3-propanediol hydrochloride, mp 258-258.5,
(C,H,N,Cl).
Additional comments: Azetropic removal of H2O is continued until
complete. Th~s process normally takes 1-4 h. If thP crude
reaction mixture does not form an eas~ly filterable solid, it can
be extracted with EtOAc (2x500 ml). The organic layers are then
combined, washed with H2O (2x500 ml), satd. NaCl solution ~2x500
ml), dried (K2CO3), filtered and the solvent removed to give the
solid crude free base which is converted to its HCl salt by the
method described above. Normally CH30H, EtOH and occasionally
i-PrOH is used to produce the solution. Alternati~ely, in cases
where the HCl salt is not soluble enough in H2O, the CH3SO3H salt
is made by treating the free base with 1.1 equivalents of CH3SO3H
AJR/JJ/6th March, 1990.
21DlZ6%6 PB1125
(99.5~, Alfa Products, P.O. Box 8247, Ward Hill, MA 01835-5747)
dissolved in abs. EtOH (10% ~/v) in an appropriate alcohol
solv0nt (CH30H,EtOH,i-PrOH). After dissolution, the mixture is
filtered thrGugh a sintered glass frit and diluted with Et20.
The resulting solid is recrystallized once or twice to give the
pure final product.
Example 2
2-Methvl-2-(((10-methyl-10H-thie ~3,2-a~carbazol-4-Yl)methvl)a~ino)-
2A 10-Methyl-10H-thieno(3,2-a)ca* azole
Using the procedure outlined in Example lC, 10_-thieno(3,2-a)
carbazole (Cambridge Chemicals, Inc., 202 East Smith S~reet,
Milwaukee, WI 53207) and dimethyl sulfate (Aldrich) gave a 96.0%
yield of 10-methyl-10_-thieno(3,2-a)carbazole. This material was
used without further purification.
2B 10-Meth~l-10H-thieno(3,2-a)carbazole-4-carbaldehyde
10-Methyl-10_-thieno~3,2-a)carbazole (2A,30.0 g,l26 mmol) was
for~ylated using the procedure of A. Rieche et al., Chem. Ber.
93, 88 (1960). The cruds reaction product was passed through a
plug of SiO2 using CH2Cl2 as the eluting solvent. The
appropriate fractions were combined and the solvent removed to
give after drying 6.04 g (18~ yield) of 10-methyl-lOH-thieno
(3,2-a)carbazole-4-carbaldehyde.
2C 2-Methyl-2-(((10-methyl-10H~thieno(3.2-a)carbaæol-4-yl~methyl~
amino~-1.3-propanediol meth n s lfonate
Using the procedure outlined in Example lE, 10-methyl-lOH-thieno-
(3,2-a)carbazole-4-carbaldehyde (2B) and 2-amino-2-methyl-1,3-
AJR/JJ/6th March, 1990.
230 lZ62 6 PB1125
propanediol (Aldrich) gave a 44.0% yield of 2-methyl-2-(((10-
methyl-lOH-thieno(3,2-a)carbazol-4~yl)methyl)amino)-1,3-propane-
diol methanesulfonate, mp 187-188.5, (EtOH/Et20), (C,H,N.S).
Example 3
2-Methyl-2-(((10-methYl-lOH-thienc(3.2-a~carbazol-2-vl~methyl~2mino~-
1,3-propanediol
3A 10-Methyl-lOH-thieno~3.2-a)carbazole-2-carbaldehxde
Using the procedure outlined in Example lD, 10-methyl-lOH-thieno-
(3,2-a)carbazole-2-carbaldehyde (2A,30.75 g,l33.8 mmol) was for-
mylated to give 36.92 g of crude solid. Trituration in ho~ PhCH3
followed by recrystallization from EtOAc/hexane (1:2) gave after
drying 10.90 g (30.7% yield) of 10-methyl-10_-thieno(3,2-a)carba-
zole-2-carbaldehyde, mp 207-208.5 (C,H,N,S).
3B 2-Methyl-2-~((10-methYl-lOH-thleno(3.2-a)carbazol-2-Yl~methYl~-
amino)-l 3-propanediol methanesulfonate
Using the procedure outline in Example lE, 10-methyl-10_-thieno
(3,2-a)carbazole-2-carbaldehyde (3A) and 2-amino-2-methyl-1,3-
propanediol (Aldr~ch) gave a S8.2~ yield of 2-methyl-2-(((10-
methyl-lOH-thieno(3,2-a)carbazol-2-yl)methyl)amino)-1,3-propane-
diol methanesulfonate, mp 219-220, (C,H,N,S).
2(((Benzofuro(5.4-b)benzofuran 8-yl2methYl)amino)-2-methyl-1~3-
ropanedlol
4A Ethyl-2-(DibenzofuranYlox~ cetate
AJR/JJ/6th March, 1990.
2~ 6~
~'B1125
- 24 -
To a 2 L 3-necked RB-flask equipped with m0chanical stirrer,
reflux condenser, addition Eunnel and N2 inlet line was added
2-hydroxydibenzofuran (Aldrich,250 g,0.814 mol), K2C03 ~Mallinck-
rodt, Inc., 2nd & Mallinckrodt Street, St~ Louis, M0 63147,247.61
g,l.79 mol) and dry acetone (~00 ml). The mixture was stirred
vigorously for 45 min. at RT. BrCH2COOEt (Aldrich 136 g,0.814
mol) was then added dropwisa to the flask over 0.5 h. The
mixture was then refluxed for 2.5 h. The m~xture was cooled to
RT, filtered and the solvent removed from the resulting filtrate
to give a dark brown semisolid. This material was stirred
vigorously with hot hexane (3 L). The light yellow solution was
decanted from a dark brown oil. The solution was decolorized
with Norit , filtered through a Celite plug, concentrated to a
volume of 2 L and allowed to stand at RT overnight. The
off-white crystals that formed were removed by filtration. A
second crop of product was obtained from the filtrate after
refrigeration overnight. The two crops were combined and dried
affording 103 g (4~.8~ yield) of crude product. This material
was used without further purification. Additional crystallation
fro~ hexane gave pure ethyl 2-(2-dib,enzofuranyloxy)acetate,
mp 51.5-53,(C,H).
4B Ethvl 2-((L-Formyl--2-dibenzofur-anyl~oxv~acetate
4C Ethyl 2-((3-Fo,rmvl-2-dibenzofuranYl)oxv)acetate
Ethyl 2-(2-dibenzofuranyloxy)acetate (4A,182.0 g, 0.673 mol) was
formylated using the procedure of A. Rieche et al., Chem. Ber.
93t 88 (19~0). The reaction mi~ture was passed through a plug of
SiO2 using CH2C12 as the eluting solvent affording after
combination of the appropriate fractions and removal of solvent
151.3 g of crude material shown to contain two isomeric aldehyde
esters by NMR. The mixture was again chromatographed on SiO2
using CH2C12 as the eluting solvent. The fractions containing
isomerically pure aldehyde esters were combined. Fractions
AJR/JJ/6th March, 1990.
PB1125
- 25 -
containing mixtures of the two aldehyde esters were also combined
and rechromatographed unt$1 the mixture was co~pletely separated.
A total of 71.3 g (3S.5% yield) of the faster eluting (Rf 0.24,
SiO2/CH2C12) aldehyde ester isomer shown by ~IR to be the l-CHO
derivative was obtained and used without further purification.
Recrystallization ~EtOAc) of a portion oi this material gave pure
ethyl 2-((1-formyl 2-dibenzofuranyl)oxy)acetate, mp 100.5-101.5,
(C,H). A total of 81.2 g (40.4~ yield) of the slower eluting
aldehyde ester isomer (Rf = 0.14,SiO2/CH2C12) shown by ~MR to be
the 3-CHO derivative was obtained and used without fur-ther
purification. Recrystallization (EtOAc) of a portion of this
material gave pure ethyl 2-((3-formyl-2-(dibenzofuranyl)-oxy)-
aceta~e, mp 177-178, (C,H).
4D EthYl Benzofuro(5.4-b~benzofuran-2-carboxvlate
~; ,
To a 1 L 3-necked RB flask equipped with mechanical stirrer,
reflux condenser, thermometer and N2 inlet line was added ethyl
2-((1-formyl-2-dibenzofuranyl)oxy)acetate (4B,5628 g,O.189 mol),
~; K2C03 (Mallinckrodt, 31.29 g,O.226 mol) and dry DNF (600 ml).
; I'he mixture was heated to 150 over 20 min. and then sti.rred for
' 1 h. The reaction was cooled to RT and filtered through a
; fritted glass funnel. The solvent was removed to give 62.1 g of
crude brown oil. ~his material was partitioned between EtOAc
(1.5 L) and H20 (l L). The EtOAc layers were combined and washed
with H20 ~750 ml), satd. NaCl (750 ml) and then dried (Na2S04).
The mixture was filtered and the solvent removed to give after
drying 54.2 g of off-white solid. The solid was dissolved in
war~ PhCH3 (400 ml) and passed through a plug of SiO2 using PhCH3
as the eluting solvent. The appropriate fractions were combined
and the solvent removed to give 35.9 g (61.7~) of ethyl benzo-
furo(5,4-b)benzofuran-2-carboxylate, mp 176.5-178.5, (C,H).
4E Ethyl 8 _ rmYl-benzoEuro(5 ~-b~benzofuran-2-carboxylate
~'
AJR/JJ/6th March, 1990.
Z~626
PB1125
- 26 -
4F Ethyl 9-Formyl-benzoflroC5~4-h)benzofuran-2-carboxylate 0.1
~1-2
4G Ethyl 5:Formvl-benzofuro(5.4-b~benzofuran-2-carboxYlate
4H E~hyl 4-Formyl-benzofuro(5.4-b)benzofuran-2~carboxvlate
Ethyl benzofuro(5,4-b)benzofuran-2-carboxylate (4D,63.2 g,
0.225 mol) was formylated using the procedure of A. Rieche et
al., Chem. Ber. 93, 88(1960). The crude reaction product was
chromatographed on SiO2 using CH2C12 as the eluting solvent
several times until the mixture was resolved into four aldehyde
esters:
Ethyl 8-formylbenzofuro(5,4-b)benxofuran-2-carboxylate, 4.64 g
t6.7% yield), mp 214-215, isolated directly from chromatography,
(C,H), Rf - 0.62 (SiO2/CH2/C12).
Ethyl 9-formylbenzofuro(5,4-b)benzo:Euran-2-carboxylate 0.1 CH2-
C12, 19.8 g (27.7~ yield), mp 176-178.5, isolated directly from
chromatography, (C,H), Rf - 0.60 (SiO2/CH2C12).
Ethyl 5-formylbenzofuro(5,4-b)benzofuran-2-carboxylate, 0.05 g
(0.07~ yield), mp 201-202.5, (PhCH3), (C,H), Rf = 0.55 (SiO2/
2 2)
Ethyl 4-formylbenzofuro(5,4-b)benzofuran-2-carboxylate, 16.6 g
(23.9~ yield), Mp 225~ (dec), (CHC13), Rf = 0.35 (SiO2/CH~C12).
4I Ethvl 8-(HydroxYmethYl~benzofuro(5.4-b~benzofuran-2-carboxYlate
To 1 L RB flask equipped with magnetic stirring bar and N2 inlet
line was added ethyl 8-formylbenzofuro(5,4-b)benzofuran-2-car-
boxylate (4E,3.70 g,l2.0 Mmol) and dry THF (600 ml). The Mixture
was warmed until the starting material was completely dissolved.
AJR/JJ/6th March, 1990.
2~
- 27 - PB1125
The solution was cooled to 0~ and NaBH4 (Aldrich, 0.45 g,l2.0
mmol) was added in one portion. After stirring for 30 mins. at
0, the reaction mixture was treated with H20 (300 ml) and
acidified to pH ~ 1 with 1 N HCl. The mixture was extracted with
EtOAc (3x500 ml), satd. NaCl solution (3xSOO ml) and dried
(NaS04). Ater filtration the solvent was removed and the
resulting white solid dried in a vacuum oven. A total of 3.80 g
(-100% yield) of ethyl 8-(hydroxymethyl)benzofuro(5,4-b)benzo-
furan-2-carboxylate, mp 172-173, (C,H) was obtained and used
directly in the next step.
4J 8-(HydroxYmeth~l)benzofuro(5.4-b2benzofuran-2-carboxvlic acid
To a 500 ml RB flas~ equipped with magnetic stirring bar and N2
inlet line was added ethyl 9-(hydroxymethyl)benzofuro(5,4-_)-
benzofuran-2-carboxylate (4I,3.70 g,il.92 mmol), TH~ (lSO ml) and
a solution of 1 N NaOH solution (12 ml,l2 mmol) in EtOH (25 ml).
The solution turns milky white instantly. After 20 min. H20 (50
ml) was added to the mixture. The resulting white solid was
filtered and washed with H20 (3x200 ml) and dried overnight in a
vacuum oven at 90 after drying 3.17 g (93.2~ yield) of 8-(hydro-
xymethyl)benzofuro(5,4-b)benzoi`uran-2-carboxylic acid, 0.2 H20,
mp 260-262 (dec), (C,H).
4K Benzofuro(5.4-b)benzofuran-8-methanol
To a 500 ml 3-necked RB flask equipped with magnetic stirring
bar, reflux condenser, thermometer and N2 inlet line was added
8-(hydroxymethyl)benzofuran-2-carboxylic acid 0.2 H20 (4J,3.13 g,
11.1 mmol), Cu20 (Aldrich, 4.76 g,33.27 mmol) and quinoline
(Aldrich, 225 ml). The reaction mixture was warmed to 200-205
and stirred for 30 min. After cooling, the quinoline was removed
by distillation leaving a dark grsen oil. This material was
dissolved in CH2C12 (150 ml) and passed through a plug of SiO2
using CH2C12 as the eluting solvent. The appropriate fractions
AJR/JJ/6th March, 1990.
PB1125
- 28 -
were combined snd the solvent removed by rotary evaporation to
give after drying, 2.26 g (85.6~ yield) of benzofuro(5,4-b)benzo-
furan-8-methanol, mp 137-138, (C,H).
4L Benzofuro(5,4-b)benzofuran-8-carbalde'hvde
To a 500 ml RB flask equipped with magnetic stirring bar, reflux
condenser and N2 inlet line was added benzofuro(5,4-b)benzofuran-
8-methanol (4K,2.26 g,0.29 ~mol), BaMnO4 (Aldrich, 4.76 g,l8.6
mmol) and CH2C12 (300 ml). The mixture was reiluxed for 2 h,
R
filtered through a plug of Celite and the solvent removed to
give after drying 2.08 g oE crude product. This material was
dissolved in CH2C12 (S0 ml) and passed through a plug of SiO2
using PhCH3 as the eluting solvent. The appropriate fractions
were combined and the solvent removed to give 2.06 g of (94.1%)
of benzofuro(5,4-_)benzofuran-8-carbaldehyde, mp 156.5-158,
(C,H)
4M 2(((Benzofuro(5.4-b~benzofuran~ methYl~amino)-2-methyl-
~_-propanediol Hvdrochloride
Usin~ the procadure outlined in Example lE, benzofuro(5,4-_)ben-
zofuran-8-carbaldehyde (4L) and 2-amino-2-methyl-1,3-propane.diol
(Aldrich) gave a 57.3% yield of 2-(~(benzofuro(5,4-b)benzofuran-
8-yl)methyl)amino)-2-methyl-1,3-propanediol hydrochloride,
mp 262-263.5, (CH3OH/Et2O), (C,H,N,Cl).
Example 5
2-(((Ben~ofuro(5,4-b)benzofuran-9-vl~methyl~amino)-2-me hyl-
5A Ethyl 9-(HYdroxymethvl~benzofuro(5.4-b~benzofuran-2-carboxylate
AJR/JJ/6th March, 1990.
PB1125
~ - 29 -
Using the procedure outlined in Example 4I, ethyl
9-formylbenzofuro(5,4-b)benzofuran-2-carboxylate 0.1 CH2C12 gave
a 88.9~o yield of ethyl 9-~hydroxymethyl)benzofuran(5,4-b)ben-
zofuran-2-carboxylatej mp 138-140, ~C,H).
5B 9-(HYdroxymethYl~benzofuro~5,4-b2benzofuran-2-carboxYlic acid
Using the procedure outlined in Example 4J, ethyl 9-(hydroxy-
methyl)-benzofuro(5,4-_)benzofuran-2-carboxylate ~5A) gave a
98.6~ yield of 9-(hydroxymethyl)benzofuro(5,4-b)benzofuran-2-
carboxylic acid, mp > 295, (C,H).
5C Benzofurant5,4-b)benzofuran-9-methanol
Using the procedure outlined in Example 4K, 9-(hydroxymethyl)-
benzofuro(5,4-_)benzofuran-2-carboxylic acid (5B) gave a 75.9
yield of benzofuro(5,4-b)benzofuran-9-methanol, mp 132-133,
(C,H).
5D Benzofuro~5,4-b~benzofuran-9-carbalclehYde
Using the procedure outlined in Example 4L, benzofuro(5,4-_)-
benzofuran-9-m0thanol (SC) gave a 92.3~ yield of benzofuro-
(5,4-b)benzofuran-9-carbaldehyde, mp 183-184, (C,H~.
5E 2-t((Benzofuro(5.4-b)benzofuran-9-Yl)methvl)amino)-2-methyl-
1,3-propanediol HYdrochloride
Using the procedure outlined in Example lE, benzofuro(S,4-b)-
benzofuran-9-carbaldehyde (5~) and 2-amino-2-methyl-1,3-propane-
diol (Aldrich) gave a 34.2~ yield of 2-(((benzofuro(5,4-b)benzo-
furan-9-yl)methyl)amino)-2-methyl-1,3-propanediol hydrochloride,
mp 243-244.5, ~EtOH/Et20), (C,H,N,Cl).
Examvle 6
AJR/JJ/6th March, 1990.
~::
PB1125
- 30 -
2-(((Benzofuro(5 4-b)benzofuran-4-Yl~methvl!amino~-2-methyl-
1 3-ero~anediol
6A 4-Formvl-benzofuro(5.4-b)benzofuran-2-carboxYlic acid
Using the procedure ou~lined in Example 4J, ethyl 4-formylbenzo-
furo(S,4-b)benzofuran-2-carboxylate (4H) gave a 81.8% yield of
4-formylbenzofuro(5,4-b)benzofuran-2-carboxylic acid which was
directly in the next step without further purification.
6B Benzofuro(5,4-b~benzofuran
6C Benzofuro(5.4-b)benzofuran-4-methanol
4-Formyl-benzofuro(5,4-b)benzofuran-2-carboxylic acid was
decarboxylated (6A,13.60 g,48.5 mmol~ using the procedure
outlined for the hydroxymethyl carboxylic acid derivative in
Example 4K. Two major products were formed in this reaction and
crudely separated by column chromatography (SiO2) using CH~C12 as
the eluting solvent. The fractions containing the faster eluting
material were collected the solvent removed and the residue
chromatographed (SiO2) using PhCH3 as the eluting solvent. The
appropriate fractions were combined and the solvent removed to
give 2.61 g (25.8~ yield) of benzofuro(5,4-_)benzofuran,
mp 66-67, C,H. The fractions containing the slouer eluting
material were also combined and the solvent removed to give
3.79 g (3~.7~ yield) of benzofuro(5,4-_)benzofuran-4-methanol),
mp 129-131, C,H. The aldehyde carboxylic acid starting
materials undergoes a Cannizaro type reaction unlike the
hydroxymethyl carboxylic acid (4J).
6D Benzofuro(5.4-b~benzofuran-4-carbaldehYde
Usin~ the procedure outlined in Example 4L, benzofuro(S,4-_)-
benzofuran-4-methanol (6C) gave a 78.6% yield of benzofuro-
~JR/JJ/6th March, 199~.
~26~
PB1125
- 31 -
(5,4-b)benzofuran 4-carbaldehydP, mp 170.5-171.5, (C,H).
6E ~ nzofuro(5,4-b)ben~ofuran-4-yl2methYl~amlno)-2-methyl-
1,3-~ro~anediol Hydrochloride
Using the procedure outlined in Example lE, benzoiuro(5,4-b)-
benzofuran-4-carbaldehyde (6D) and 2-amino-2-methyl-1,3-propane-
diol (Aldrich) gave a 52.0~ yield of 2-(((benzofuro(5,4-b)b~nzo-
furan-4-yl)methyl)amino)-2-methyl-1,3-propanediol hydrochloride,
mp 263-265 (dec), (CH3OH/Et20), (C,H,N,Cl).
Exam~l~_Z
2-(((Benzofuro(5,4-b~benzofuran-2-yl)methyl)amino)-2-methyl-
1,3-propanediol
7A Benzofuro(5,4-b)benzofuran-2-carbaldehyde
To a 300 ml RB flask equipped with magnetic stirring bar, rubber
septum and N2 inlet line was addPd benzofuro(S,4-_)benzouran
(6B,2.46 g,ll.81 mmol), dry THF (125 ml) and N,N,N',N'-tetra-
methylethylenediamine (Aldrich, 3.16 g,27.17 mmol,4.10 ml) (dis-
tllled from CaH2). The mixture was cooled to -7B. s-BuLi
(Aldrich, 1.3 M in cyclohexane, 27.17 mmol, 20.9 ml) was added
. dropwlse to the mixture by syringe. The mixture was stirred at
; -78 for 1 h, warmed to -20 for 45 min. and then recooled to
-78. Dry DMF (Mallinckrodt, 3.20 g,41.3 mmol,3.1 ml) was then
added dropwise by syringe to the flask. The mixture was stlrred
at -78 for 10 min. and then allowed to warm to RT (over 1 h).
The reaction was quenched with H2O (25 ml) and acidified with 1 N
HCl. The THF was removed by rotary evaporation and the residue
further diluted with H2O (300 ml). This material was extracted
with EtOAc (3x500 ml). The organic layers were combined and
washed with H~O (2x500 ml), satd. NaCl solutlon (1 L) and dried
(Na2SO4). The solvent was removed to give 2.49 g of yellow
AJR/JJ/6th March, 1990.
PB1125
- 32 -
solid. This material was chromatographed (SiO2) using PhCH3 as
the eluting solvent. The appropriate fractions were combined and
the solvent removed to give 1.52 g (54.5~ yield) of benzofuro-
(5,4-b)benzofuran-2-carbaldehyde, mp 193-194.5, (C,H).
7B 2~ enzofuro(5,4-b~benzofuran-2-yl~methyl~amino~-2-meth
1,3-propanediol hvdrochloride
Using the procedure outlined in Example lE, benzofuro(5,4-b)
benzofuran-2-carbaldehyde (7A) and 2-amino-2-methyl-1,3-propane~
diol (Aldrich) gave a 41.5~ yield of 2-(((benzofuro(5,4-b)benzo-
furan-2-yl)methyl)amino)-2-methyl-1,3-propanediol hydrochloride,
mp 230-230.5 (dec), (EtOH/Et20), (C,H,N,Cl).
Example 8
2-(~(Benzofuro(5~6-b~benzofuran-8-yl~methYl)Amino~-2-methYl-
1.3-propanediol
8A Ethy~ Benzofuro(5~6-b)benzofuran-2-carboxvlate
To a 1 L 3-necked RB flask equipped with mechanical stirrer,
reflux condenser, thermometer and N2 inlet line was added ethyl
2-((3-formyl-2-dibenzofuranyl)oxy)acetate (4C,67.32 g,0.225 mol),
K2CO3 (Mallinckrodt, 37.43 g,0.271 mol) and dry DMF (800 ml).
The mixture was heated to 155~ for 1.5 h. The reaction was
cooled to RT and filtered through a fritted glass funnel. The
solvent was re~oved to give 82.1 g of crude brown oil. This
material was partitioned between EtOAc (1.5 L) and H2O (2x750
ml), satd. NaCl (2xl L) and then dried (Na2S04). The mixture was
filtered and the solvent removed to give after drying 53.2 g of a
yellow solid. The solid was dissolved in warm PhCH3 (550 ml) and
passed through a plug of SiO2 using PhCH3 as the elutlng solvent.
The appropriate fractions were combined and the solvent removed
to give 40.4 g (53.2% yield) of material that was used without
AJR/JJ/6th March, 1990.
PB1125
iEurther purification. Recrystallization oiE a small amount of
material from PhCH3 gave ethyl benzofuro(5,6-b)benzofuran-2-
carboxylate, mp 123.5-124.5, (C,H).
8B Ethyl 8-Formyl-benzofuro(5~6-b~benzofuran-2-carboxylate 0.05
8C EthYl 4-Formyl-benzofuro(5,6-b)benzofuran-2-carboxylzte
Ethyl benzofuro(5,6-_)benzofuran-2-carboxylate (8A,36.0 g, 0.132
mol) was formylated using the procedure of A. Rieche et al.,
Chem. Ber. 93, 88(1960). ~he crude reaction ~ixture was passed
through a plug of SiO2 using CH2C12 as the elutiLng fraction
several times until the mixture was resolved into two aldehyde
esters:
Ethyl 8-formylbenzofuro(5,6-k)benzofuran-2-carboxylate 0.05
CH2C12, 6.18 g (15.2~ yield), mp 227, isolated directly from
chromatography, (C,H), Rf ~ 0.23 (SiO2/CH2C12);
Ethyl 4-formylbenzofuro(5,6-k)benzoiEuran-2-carboxylate, 26.2 g
: (64.4~ yield), (PhCH3), (C,H) Rf 0-63 (Sio2/CH2C12).
8D Ethyl 8-~HvdroxYmethvl~benzofuro(5,6-b~benzofuran-2-carboxvlate
Using the procedure outlined in Example 4I, ethyl 8-formyl-benzo-
; furo(5,6-k)benzofuran (8C) gave a 93.3~ yield of ethyl
8-(hydroxymethyl)benzofuro(5,6-k)benzofuran-2-carboxylate,
~ mp 186-186.5~, (C,H).
; 8E 8-(HydroxYmethvl)benzofuro~5.6-b?benzoiEuran-2-carboxvlic acid
Using the procedure outlined in Example 4J, ethyl 8-(hydro-
AJR/JJ/6th March, 1990.
2~Z6~
PB1125
- 34 -
methyl)benzofuro(5,6-b)benzofuran-2-carboxylate (8D) gave a 92.7%
yield of 8-~hydroxymethyl)benzofuro(5,6-b)benzoiuran-2-carboxylic
acid, mp >270, (C,H).
8F ~ Benzofuro(5 6-b)benzofuran-8-methanol
Using the procedure outlined in Example 4K, 8-(hydroxymethyl)-
benzofuro(5,6-b)bPnzofuran-2-carboxylic acid (8E) gave a 74.8
yield of benzofuro(5,6-b)benzofuran-8-methanol, mp = 170.172,
(C,H).
8G Benzofuro(5.6-b~benxofuran-8-carbaldehy~
Using the procedure outlined in Example 4L, benzofuro(5,6-b)-
benzofuran-8-methanol gave a 89.7% yield of benzofuro(5,6-_)-
benzofuran-8-carbaldehyde, mp 188-189, (C,H~.
8H 2t((Benzofuro(5.6-b~benzofuran-8-Yl~methyl~amino)-2-methYl-
1,3-~ro~anediol HYdrochloride
Using the procedure outlined in Example lE, benzofuro(5,6-_)-
benzofuran-8-carbaldehyde (8G) and 2-amino-2-methyl-1,3-propane-
diol (Aldrich) gave a 66.3~ yield of 2-(((benzofuro(5,6-k)benzo-
furan-8-yl)methyl)amino)-2-methyl-1,3-propanediol hydrochloride,
mp 235-236.5 (dec), (CH3OH~Et2O), (C,H,N,Cl).
Exam~le 9
2-(((Benzofuro(5 6-b~benzoiuran-4-yl)methyllamino)-2-methvl-
1,3-propanediol
9A 4-Formvl-benzofuro(5.6-b~benzofuran-2-carboxYlic acid
IJsing the procedure outlined in Example 4J, ethyl 4-formyl-benzo-
furan-2-carboxylic (8C) gave a 98.9~ yield of 4-formyl-benzofuro-
AJR/JJ/6th March, l990.
2~
PB1125
35 -
(5,6-_3benzofuran-2-carboxylic acid, that was used directly in
the next step without further purification.
9B Benzofuro(5 6-b~benzofuran
9G Benzofuro(5.6-b~benzofuran-4-methanol
4-Formyl-benzofuro(5,6-b)benzofuran-2-carboxylic acid (9A,25.2 g,
81.7 mmol) using the procedure outlined for the hydroxymethyl
carboxylic acid derivatlve in Example 4K. The two ma;or products
formed in this reaction were crudely separated by rolumn
chromatography (SiO2) using CH2C12 as the eluting solvent. The
fractions containing the faster eluting material were collected,
the solYent removed and the residue chromatographed (SiO2) using
PhCH3 as the eluting solvent. The appropriate fractions were
combined and the solvent removed to give 6.20 g (33.4~ yield) of
benzofuro(S,6-_)furan, mp 90-100, (C,H). The fractions
containing the slower eluting materlal wsre also combined and the
solvent removed to give 3.91 g (18.4% yield) of benzofuro(5,6-_)-
benzofuran-4-methanol., mp 152-153, (C,H). As mentioned for
Examples 6B and 6C, the aldehyde carboxylic acid starting
material undergoes a Cannizaro type reaction unlike the
hydroxymethyl carboxylic acid 8E.
: "
9D Benzofuro(5.6-b)benzofuran-4-carbaldehYde
Using the procedure outlined in Example 4K, benzofuro(5,6-b)-
benzofuran-4-methanol (9C) gava a 96.5~ yield of benzofuro-
(5,6-b)benzofuran-4-carbaldehyde, mp 167.5-169, (C,H).
9E 2-(((Benzofuro(5 6-b)benzofuran-4-Yl)methYl)amino)-2-methyl
1.3-propanediol HYdrochloride
Using the procedure outlined in Example lE, benzofuro(5,6-b)-
benzofuran-4-carbaldehyde (9D) and 2-amino-2-methyl-1,3-propane-
AJR/JJ/6th March, 1990.
2~
PB1125
- 3~ -
diol (Aldrich) gave a 46.9% yleld of 2-(((benzofuro(5,6-~)-
benzofuran-4-yl)methyl)amino)-2-methyl-1,3-propanediol
hydrochloride, mp 224-225 (dec), (EtQH/Et20), (C,H,~,Cl).
Example 10
2-(t(Benzofurot5~6-h~benzofuran-2-yl)methyl~amino~-2-methYl-
1~3-propanediol
lOA Benzofuro(5,6-b)benæofuran-2-carbaldehyde
Using the procedure outlined in Example 7A, benzofuro(5,6-b~-
benzofuran (9B) gave a 82.8~ yield of benzofuro(5,6-b)benzofuran-
2-carbaldehyde, mp 194-195.5, (C,H~.
lOB 2-((tBenzofuro(5~6-~benzofuran-2~yl~methYl~amino~-2-methYl-
1 3-propanediol Hydrochloride
Using the procedure outlined in Example lE, ban~ofuro(5,6-b-)
benzofuran-2-carbaldehyde (lOA) and 2-amino-2-amlno-2-methyl-
1,3-propanediol (Aldrich) ga~e a 50.3% yield of 2-(((benzofuro-
(5,6-_)benzofuran-2-yl)methyl)amino)-2-methyl-1,30-propanediol
hydrochloride, mp 231-232.5 (dec), (CH30H/Et20), (C,H,N,Cl).
ExamDle 11
2-Methyl-2-t((l-methyl-lH-Lllbenzothieno~2~3-~lindol-3
methyl~amino~ 3--e,ropanediol
llA EthYl l-Methyl-lH~llbenzothieno~2,3-glindole-2-carboxYlate
Using the procedure outlined in Example lC, ethyl lH[l]benzo-
theino-[2,3-g]indole-2-carboxylate (H.G. Pars Pharmaceutical
Laboratories, Inc., 763 Concord Ave., Cambridge, MA 02133) and
AJR/JJ/6th March, 1990.
~2~
PB1125
- 37 -
dimethyl sulfate (Aldrich) gave a quantitative yield of ethyl
l-methyl-lH-[l]benzothieno[2,3-g]-indole-2-carboxylate,
mp 85-86, (PhCH~), (C,H,N).
llB Ethyl_3-Formyl-2-methvl-1H[llbenzothieno~2.3-~lindole-2
carboxvlat_
Ethyl l-methyl-lH-[l]benzotheino[2,3-g]indole-2-carboxylate (llA)
was formylated by the procedure of A. Rieche et al., Chem. Ber.
93, 88~1960) to give a 58.7~ yi~ld of ethyl 3-formyl-1-methyl-lH-
[l]benzothieno[2,3-g]indole-2-carboxylate, mp 145-147, (CH2C12/
hexane), (C,H,N).
llC 3-Formvl-l-methvl-lH-rllbenz_thieno~2 3-~lindole-2-carboxvlic
acid
Using the procedure outlined in Example 4J, ethyl 3-formyl-1-
methyl-lH-[l]benzothieno[2,3-g]indole-2-carboxylate (llB) gave a
91.0% yield of 3-formyl-1-methyl-1_-[1]-benzothieno[2,3-g]indole-
2-carboxylic acid, which was used directly without further
purification.
llD l-Methyl-lH-~ll-benzothieno~2.3-glirldole-3-carbaldehvde
Using the procedure outlined in Example 4K, 3-fGrmyl-l-methyl-
lH-[l]-benzothieno[2,3-g]indole-2-carboxylic acid (llC) gave a
35.3~ yield of 1-methyl-lH-benzothieno[2,3-g]indole-3-carbalde-
hyde, mp 192-194, (CH2C12/petroleum ether), (C,H,N).
llE 2-Methyl-2-(((1-methvl-lH-~llbenzothieno~2 3-~lindol-3-vl)
methvl)amino)-1.3-propanediol HYdro_ loride
Using the procedure outlined in Example lE, l-methyl-lH-[l]-
benzothieno-[2,3-g]-indole-3-carbaldehyde (llD) and 2-amino-2-
methyl-1,3-propanediol (Aldrich) gave a 56.7~ yield of 2-methyl-
AJR/JJ/6th March, 1990.
20~Z62G
PB1125
- 38 -
2-(((1-methyl-lH-[l]benzothieno[2,3-~]indol-3-yl)methyl)amino)-
1,3-propanediol hydrochloride, mp 228-230 (dec), (CH30H/Et20),
(C,~,~,Cl).
Example 12
2-l((6-Ethyl-1 6-dihvdro-1-methvlpyrrolo~3 2-clcarbazol-3-yl~-
methyl~amino)-2-methvl-1,3-propanediol
12A Ethvl 6-Ethyl-l,lL~bL~y_r~o-l-methYlpyrrolo[3.2-c]carbazole-2-
carboxylate
Uslng the procedure outlined in Example lC, ethyl 6-ethyl-1,6-
dihydro-pyrrolo[3,2-c]carbazole-2-carboxylate (H.G. Pars Pharma-
- ceutical Laboratories, Inc.) gave a 90.5% yield of ethyl 6-ethyl-
1,6-dihyclro-1-methylpyrrolo[3,2-c]carbazole-2-carboxylate,
mp 115.5-116D, (CH2C12/hexane), (C,H,N).
12B 6-EthYl-1.6-dihYdro-l-methylpYrrolo~3.2-clcarbazole-3-
carbaldeh~de
Using the procedures outlined in Examples 4H, 4J and 4K, ethyl
6-ethyl-1,6-dihydro-2-methylpyrroio[3,2-c]carbazole-2-carboxylate
(12A) gave a 9.3% yield of 6-ethyl-1,6-dihydro-1-methylpyrrolo-
[3,2-c]carbazole-3-carbaldehyde, mp.l44-146, (CH2C12), (C,H,N).
12C 2-(((6-Ethyl-1.6-dihYdro-l-methYlpYrrolo~3.2-c~carbazol-3-yl)
methyl)-amino)~2-methy~1.3-propanediol Hydrochloride 0.6 H20
. Using the procedure in Example lE, 6-ethyl-1,6-dihydro-1-methyl-
- pyrrolo[3,2-c]carbazole-3-carbaldhyde (128~ and 2-amino-2-methyl-
1,3-propanediol (Aldrich) gave a 48.3% yield of 2-[[6-ethyl-1,6-
dihydro-l-methylpyrrolo[3,2-c]carbazol-3-yl)methyl]amino]-2-
methyl-1,3-propanediol hydrochloride 0.6 H20, mp 265-267,
(EtOH/Et20), (C,H,N,Cl).
AJR/JJ/6th March, l990.
PB1125
- 39 -
Exam~le 13
2-Methvl-2-~((1-me~hvl-lH-benzofuro[2.3-~lindol-3-vl~methyl)-amino)-
1 3-propanediol
13A Ethvl l-Meth~l-lH-benzof.uro~2,3-gLindole-2-carboxvlate
Using the procedure outlined in Example lC, ethyl lH-benzofuro-
[2,3-g]indole-2-carboxylate (H.G. Pars Pharmaceutical Laborato-
ries, Inc.) gave a ô6.3% yield oE ethyl l-methyl-l_-benzofuro-
[2,3-g]indole-2-carboxylate, mp 114-116, (EtOAc), (C,H,N).
13B Ethvl 3-Formvl-l-methYl-lH-benzofuro r 2.3-ylindole-2-carboxvlate
Ethyl l-methyl-lH-benzofuro[2,3-g]indole-2-carboxylate (13A) was
formyla~ed according to the procedure of A. Rieche et al., Chem.
Ber. 93, 88(1960) to give and 82.5% yield of ethyl 3-formyl-1-
methyl-lH-benzofuro[2,3-c]-indole-2-carboxylate, mp 190-192,
(CH2C12/hexane), (C,H,N).
13C 3-Formvl-l-methvl-lH-benzofuror2.3-~ indole-2-carboxvlic acid
Using the procedure outlined in Exa~ple 4J, ethyl 3-formyl-1-
methyl-lH-benzofuro[2,3-g]indole carboxylate (13B) gave a 90.3~
yield of 3-formyl-1-methyl-1_-benzofuro[2,3-~]indole-2-carboxylic
acid, which was used directly without further purification.
13D l-Methvl-lH-benzofurol2.3-glindole-3-carbaldehyde
Using the procedure outlined in Example 4K, 3-formyl-1-methyl-lH-
banzofuro[2,3-g]indole-2-carboxylic acid (13C) gave a 40.1~ yield
of l-methyl-lH-benzofuro[2,3-g]indole-3-carbaldehyde, mp 163-
165, (CH2,C12/petroleum ether~, (C,H,N).
AJR/JJ/6th March, 1990.
20~2626 PB1125
- 40 -
13E 2-Methyl-2-((~l-methyl-lH-benzofuro~2.3-~lindole-3-vl~methy
amino2-1,3-propanediol Hydrochloride
Using the procedure outlined in Example lE, l-methyl-lH-benzo-
furo[2,3-g]-indole-3-carbaldehyde (13D) and 2-amino-2-methyl-1,3-
propanediol ~Aldrich) ga~e a 63.9% yield of 2-methyl-2-((~1-
methyl-lH-benzvfuro-[2,3-g]indol-3-yl)methyl)amino)-1,3-propane-
diol hydrochloride, mp 234-236, (EtOH/Et20), (C,H,N,Cl).
Example 14
2-Methyl-2-(((3-methyl-3H-_~llbenzothieno~2.3-elindol-1-yl3-methyl)
amino3-1,3-propanediol)
14A 3-Methvl-3H-~llbenzothieno~2.3-elindQle-l-carbaldehyde 0.25 H20
Using the procedure outlined in Example lC, ethyl 3H-ll]benzo-
thieno[2,3-e]indole-2-carboxylate (H.G. Pars Pharmaceutical
Laboratories, Inc.) and dimethyl su:Lfate (Aldrich) gave a nearly
quantitative yield of crude ethyl 3-methyl-3H-[l]benzothieno-
[2,3-e]indole-2-carboxylate. This material was formylated using
the procedure of A. Rieche et al., Chem. Ber. 93, 88(1960) to
give crude mixture of aldehydes which was hydrolyzed and
d~carboxylated without purification using the procedure outlined
in Example 15B to give a 19.0% yield of 3-methyl-3H-[l]benzo-
thieno[2,3-e]indole-1-carbaldehyde 0.25 H20, mp 223.5-224, (CH2-
C12/hexane), (C,H,N)-
14B 2-Methvl-2- U(3-methYl-3H-~llbenzothieno~2,3-el~ndol-l-vl3methvl~
amino3-1.3-pro~anediol Hvdrochloride
Using the procedure described in Example lE, 3-methyl-3~-[I]-
benzothieno[2,3-e]indole-1-carbaldehyde (14A) and 2-amino-2-
methyl-1,3-propanediol (Aldrich) gave a 41.4% yield of 2-methyl-
2-(((3-methyl-3_-[l]benzothieno[2,3-e]indol-1-yl)methyl)amino)-
AJR/JJ/6th March, 1990.
2~11Z~;Z6 PBl125
- 41 -
1,3-propanediol hydrochloride, mp 228-230, (CH30H/Et20),
(C,H,N,Cl).
Example 15
2-(((10-Methyl-lOH-l llbenzothieno~3~2-blindol-6-yl)methyl)amino~-
2-methvl-1 3-propanediol
15A 10-Methyl-lOH.-~.llbenzothieno~3.2-blindole
Using the procedure outlined in Example lC, lOH-[l]benzothieno-
[3,2-_]indole (prepared by the method of K.E. Chippendale and B.
Iddon, JCS Perkln Trans. I 2023 (1972) gave and dimethyl sulfate
(Aldrich) a 91.9% yield of 10-methyl-lOH[l]benzothieno[3,2-_]
indole, mp 175-176, benzofuro(5,6-b)benzofuran-2-carbaldehyde.
15B 10-Methyl-lOH-~llbenzothienoL3.2-blindole-6-carbaldehyde 0,2 H20
15C 10-Methyl-lOH-~llbenzothieno~3,2-blindole-3-carbaldehYde
10-Methyl-10_-[l]benzothieno[3,2-b]indole (15A) was formylated
according to the procedure of A. Rieche et Al ., Chem. Ber. 93, 88
(1960) to give a low yield of a mixture of two aldehydes.
Chromato~xaphy (SiO2/PhCH3) gave a 6.7~ yield of lO-methyl-lOH-
[l]benzothieno-[3,2-b]indole-6-carbaldehyde 0.1 H20 (15B),
mp 128-130, (CH2Cl2/hexane), (C,H,N,S), Rf = 0.39 (SiO2/PhCH3)
and a 22.3~ yield of 10-methyl-lOH-[l~benzothieno[3,2-b]indole-
3-carbaldehyde (15C), mp 188-189.5, (PhCH3/hexane), (C,H,N,S),
Rf - 0.11 ~SiO2/Ph~H3).
15D 2-(((10-MethYl-lOH-~llbenzothieno~3.2-blindol-6-yl~methvl)amino~-
2-methyl-1.3-proPanediol Hvdrochloride
Using the procedure outlined in Example lE, 10-methyl-lOH-[l]ben-
zothieno(3,2-b]indole~6-carbaldehyde (15B) and 2-amino-2-
AJR/JJ/6th March, 1990.
~ PB1125
- 42 -
methyl-1,3-propanediol (Aldrich) gave a 26.9~ yield of 2-(((10-
methyl-lOH-[l]benzothieno-[3,2-b]indol-6-yl)methyl)amino)-2-
me~hyl-1,3-propanediol hydrochloride, mp 249.5-250, (CH30H/
Et20), (C,H,N,S,CI).
Example 16
2-(((10-Methvl-lOH-~llben~othieno~3~2-blindol-3-vl)methyl~amino~-
2-methvl-1~3-propanediol hYdrochloride
Using the procedure outlined in Example lE, 10-methyl-lOH-[l]benzo-
thieno[3,2-b]indole-3-carbaldehyde (lSC~ and 2-amino-2-methyl-1,3-
propanediol (Aldrich) gave a 39.4% yield of 2-(~(10-methyl-lOH-
[l]benzothieno[3,2-b]indol-3-yl)methyl)amino)-2-methyl-1,3-propanediol
hydrochloride, mp 285.5-259.5, (CH30H/ET20), (C,H,N,S,Cl).
Antitumor Scr enine Results
Methods for evaluating the antitumor activity of these compounds are
essentially those used in the Tumour Panel by the Development
Therapeutics P~ogram, Division oi Cancer Treatment, National Cancer
Institute, A. Goldin, et al., ~ethods in Cancer Research, Vol. XVI,
p.165, Academic Press (1979). Some modifications, in dose level and
schedule have been made to increase the testing efficiency.
Lvmphocytic Leukemia P388/0 Test
CD2-Fl mice, of the same sex weighing within 20+3g, are used for this
test. Control and test animals are in~ected intraperitoneally with a
suspension of 10 viable P388/0 tumour cells on day 0. In each test
several dose levels which bracket the LD20 for the compound are
evaluated; each dose level group contains 6 animals. The test
compounds are prepared either in physiolo~ic saline containing 0.05%
Tween 80 or distilled water containing 5~ dextrose and are
administered intraperitoneally on days, 1, 5, and 9 relative to tumor
~.,
AJR/JJ/6th March, 1990.
'
2~12626 PB1125
- 43 -
implant. Doses are on a mg/kg basis according to individual aniMals'
body weights. the day of death for each animal is recorded, the
median identiiied for each group and the ratios of median survival
time for treated (T)/control (C~ groups are calculated. The criterion
for activity is T/C x 100 2120%. Results of P388/0 testing are
summarl~ed in Table I below.
TABLE I
Activity of Compounds Against P388 Lymphocytic Leukemia
Compounds LD20Optimal Dose ~ T/C* 30-Day
of Formula (mg~kg)(mg/kg) Survivors
lE 150 250 210 0/6
2C 375 350 195 0/6
3B 375 275 195 0/6
4M ~450 550 117 0/6
5E 175 150 117 0/6
6E 60 45 213 1/6
7B 175 175 138 0/6
8~ 300 300 200 0/6
9E 75 75 192 0/6
10B >675 600 100 0/6
llE ~50 300 135 0/6
12C 100 100 113 0/6
13E 115 100 120 0/6
14B 150 125 215 0/6
lSD 225 300 140 5/6
16 250 250 170 0/6
.
*Excluding 30 day survivors
:
'';
AJR/JJ/6th March, 1990.
,
. .
21~6~:~
PB1125
- 44 -
Formulation Examples
TABLET
Compound of Formula I (as hydrochloride) 500.0 mg
Pregelatinised Corn Starch 60.0 mg
Sodium Starch Glycolate 36.0 mg
Magneslum Stearate 4.0 mg
The Compound of formula (I) is finely ground and intimately mixed with
the powdered excipients, pregelatinised corn starch and sodium starch
glycolate. The powders are wetted with purified water to form
granules. The granules are dried and mixed with the magnesium
stearate. The formulation is then compressed into tablets weighing
approximately 600 mg each.
B. TABLET
Compound of formula (I) 500.0 mg
Corn Starch 70.0 mg
Lactose 83.8 mg
Magnesium Stearate 4.2 mg
Polyvinylpyrrolidone 14.0 mg
Stearic Acid 28.0 mg
The Compound of formula (I) is finely ground and intimately mixed with
the powdered excipients, corn starch and lactose. The powders are
wetted with a solution of polyvinylpyrrolidone dissolved in purified
water and denatured alcohol to form granules. The granules are dried
and mixed with the powdered stearic acid and magnesium stearate. The
AJR/JJ/6th March, 1990.
:. . .. . .
Z~262~
PB'125
formulation is then compressed in~o tablets weighing approximately 700
mg each.
C. CAPSULES
Compound of formula (I) 500.0 mg
Corn Starch 50.0 mg
Magnesium Stearate 3.0 mg
The finely divided compound of formula (I) is mixed with powdered corn
starch and wetted with denatured alcohol to densify the powder. The
dried powder is mixed with stearic acid and filled into hard-shell
gelatin capsules.
D. SYRUP
Compound of formula (I) 250.0 mg
Ethanol 250.0 mg
Glycerin 500.0 mg
Sucrose 3,500.0 mg
Flavouring Agent q.s.
Colouring Agent q.s.
Preserving Agent 0.1%
Purified Water q.s. to 5.0 ml
The Compound of formula (I) is dissolved in the ethanol, glycerin, and
a portion of the purified water. The sucrose and preserving agent are
dissolved in another portion of hot purified water, and then the
colouring agent is added and dissolved. The two solution are mixed
and cooled before the flavouring agent is added. Purified water is
added to final volume. The resulting syrup is throughly mixed.
AJR/JJ/6th March, 1990.
PB1125
- 46 -
E. IV INJECTION
Compound of formula (I) 5.0 mg
Slycerin q.s. for isotonicity
Preservative 0.1%
Hydrochloric Acid or Sodium Hydroxide as needed for pH
adjustment
Water for Injection q.s. to 1 ml
The compound of formula (I) and preservative is added to the glycerin
and a portion of the watæ.r for injection. The pH is adjusted with
hydrochloric acid or sodium hydroxide. Water for injection is added
to final volume and solution is complete after thorough mixing. The
solution is sterilised by filtration through a 0.22 micrometer
membrane filter and aseptically filled into sterile 10 ml ampoules or
vials.
AJR/JJ/6th March, 1990.
~ ' ' ': , '
