CA2012626A1 - Hetero polycyclic biocidal compounds, their synthesis and intermediates therefor, formulations containing them, and their use in medicine - Google Patents

Abstract

ABSTRACT

Heterotetracyclic aromatic compounds containing two hetero atoms selected from oxygen, sulphur and nitrogen and that are substituted by an aminoalkanol group that contains between two and eight carbon atoms are disclosed as having biocidal activity. Processes for preparing the compounds, intermediates in their preparation, pharmaceutical compositions containing them and their use in medicine are also disclosed.

6th March, 1990

Description

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.

~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 (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.

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 wherein R17 is hydrogen or methyl and Rl is hydrogen, methyl or ethyl, preferably methyl.

AJR/JJ/6th March, 1990.

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-, 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.

,:
.- . , , ~
.
..

:

- 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.

, - 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.

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.

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.

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.

- 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 - 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.

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.

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.

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.

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.

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.

- 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.

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.

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.

. .

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.

,, , :

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 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 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.

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 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~

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.

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.

~ - 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.

~::

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~

(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.

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.

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~

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~

(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~

- 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~

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

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.

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 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 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 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 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 4M ~450 550 117 0/6 8~ 300 300 200 0/6 10B >675 600 100 0/6 llE ~50 300 135 0/6 lSD 225 300 140 5/6 .
*Excluding 30 day survivors :
'';
AJR/JJ/6th March, 1990.

, . .

21~6~:~

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.

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.

~ ' ' ': , '

Claims (10)

1. 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 total, or an ester or a salt thereof;
wherein Ar is a fused tetracyclic hetero aromatic ring system of the formula:

in which Z is oxygen, sulphur or a group NRl wherein R1 is hydrogen, methyl or ethyl and is a bicyclic aromatic ring system comprising a phenyl ring and a 5-membered ring system which contains one heteroatom Z1 selected from oxygen, sulphur or a group NR2 wherein R2 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; C1-4 alkyl or C1 4 alkoxy, each optionally substituted by hydroxy or C1 2 alkoxy; halogen substituted C1-2 alkyl or C1 2 alkoxy; a group S(O)nR3 wherein n is an integer 0,1 or 2 and R3 is C1-2 alkyl optionally substituted by hydroxy or C1-2 alkoxy; or Ar is 6th March, 1990 optionally substituted by a group NR4R5 containing not more than 5 carbon atoms wherein R4 and R5 are the same or different and each is a C1 3 alkyl group or NR4R5 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 or wherein m is n or 1;
R6 is hydrogen or C1 3 alkyl optionally substituted by hydroxy;
R7 and R8 are the same or different and each is hydrogen or C1 3 alkyl;
R9 is hydrogen, methyl or hydroxymethyl;
is a five-or six-membered saturated carbocyclic ring;
R10, R11 and R12 are the same or different and each is hydrogen or methyl;

R13 is hydrogen, methyl, hydroxy, or hydroxymethyl; R14 is hydrogen, methyl, hydroxy or hydroxymethyl.

2) A compound according to claim 1 in which the tetracyclic aromatic ring system Ar is unsubstituted.

3) A compound of the formula (I) according to either claim 1 or 2 in which R is a group:

6th March, 1990 or wherein R15 is CH2OH, CH(CH3)OH or CH2CH2OH, R16 is hydrogen, C1-3 alkyl or CH2OH, and R17 is hydrogen or methyl.

4) A compound of the formula (I) according to any one of claims 1 to 3 in which R is a group:

wherein R17 is hydrogen or methyl and R18 is hydrogen, methyl or ethyl.

5) A compound of the formula (I) according to any one of claims 1 to 4 in which Ar is a group:
, or AJR/EB/6th March, 1990

6) A compound of the formula (I) according to any one of claims 1 to 5 selected from 2-(((10-Methyl-10H-[1]benzothieno[3,2-b]indol-3-yl)methyl)-amino)-2-methyl-1,3-propanediol, 2-(((lo Methyl-10H-[l]benzothieno[3,2-b]indol-6-yl)methyl)amino)-2- methyl-1,3-propanedioll 2-Methyl-2-(((10-methyl-10H-benzofuro(3,2-b)indol-6-yl)methyl) amino)1,3-propanediol, 2-(((Benzofuro(5,6-b)benzofuran-4-yl)methylamino)-2-methyl-1,3-propanediol, 2-(((Benzofuro(5,6-b)benzofuran-8-yl)methyl)amino)-2-methyl-1,3-propanediol, 2-(((Benzofuro(5,6-b)benzofuran-2-yl)methyl)amino)-2-methyl-1,3-propanediol, 2-(((Benzofuro(5,4-b)benzofuran-4-yl)methyl)amino)-2-methyl-1,3-propanediol, 2-(((Benzofuro(5,4-b)benzofuran-9-yl)methyl)amino)-2-methyl-1,3-propaned;ol, 2-(((Benzofuro(5,4-b)benzofuran-8-yl)methyl)amino)-2-methyl-1,3-propanediol, 2-(((Benzofuro(5,4-b)benzofuran-2-yl)methyl)amino-2-methyl-1,3-propanediol, and monomethyl or monethyl ethers, esters, and addition salts thereof.

7) A process for the preparation of a compound of the formula (I) according to any one of claims 1 to 6 which comprises:

(i) the reduction of a compound Ar-CH=NR (II) or an appropriately protected derivative thereof followed by deprotection where appropriate.

6th March, 1990 (ii) the reduction of a compound ArCO.NHR or a derivative thereof in which the hydroxy groups are protected, followed by deprotection where appropriate, or (iii)the reaction of a compound ArCH2L with a compound NH2R
wherein L is a leaving group; wherein Ar and R are as hereinbefore defined.

8) A novel chemical intermediate involved in the preparation of a compound of the Formula (I).

9) A compound of the formula (I) according to any one of claims 1 to 6 For use in medicine.

10) A pharmaceutical formulation which comprises a compound of the formula (I) according to claim 1 together with a pharmaceutically acceptable carrier thereof.

6th March, 1990