Tumor necrosis factor production inhibitors

There is provided a composition for inhibiting the production or secretion of tumor necrosis factor effective for the treatment of cachexia, septic shock, multiple organ failure, rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, osteoarthritis, Behcet disease, systemic lupus erythematosus (SLE), graft versus host disease (GvHD), malaria, acquired immune deficiency syndrome (AIDS), meningitis, hepatitis and Type II diabetes mellitus. The composition comprises a pharmaceutically effective amount of a compound of formula (1).

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
The present invention relates to a new medical use of a compound for 
inhibiting the production or secretion of a tumor necrosis factor. 
2. Related Art 
A tumor necrosis factor (hereinafter abbreviated as TNF) is a peptide of 
157 amino acids, having a molecular weight of about 17,000. TNF is one of 
cytokines produced by various cells including macrophages. 
TNF had been firstly found out as a cytokine showing a cytotoxic effect on 
tumor. The recent studies have revealed that the activities of TNF are not 
only limited to tumor cells but also extended to many other normal cells. 
Examples of such TNF activities include suppression of the lipoprotein 
lipase activity in adipocytes, expression of HLA antigen on blood 
endothelial cells and fibroblasts, interleukin-1 production by fibroblasts 
or macrophages, activation of cytotoxic macrophages, suppression of CFU, 
production of colony stimulating factor by fibroblasts, endothelial cells 
or some tumor cells, inhibition of the synthesis of proteoglycans and 
stimulation of their resorption in cartilage, activation of neutrophils 
and generation of superoxide, production of procoagulant factor by blood 
endothelial cells, proliferation of fibroblasts, change in membrane 
potential of skeletal muscle, interferon .beta..sub.2 production by 
fibroblasts, and injury of blood endothelial cells. In these days, TNF has 
thus been recognized to be a cytokine which takes part broadly in vital 
protection through inflammation and immune response [Vassalli, P., Ann. 
Rev. Immunol., 10, 411-452 (1992)]. 
On the other hand, it is noted that continuous or excessive production of 
TNF rather results in vigorous actions on normal cells to cause various 
diseases. For example, TNF is also known as cachectin which induces 
cachexia in cancer or infectious diseases, involving catabolic 
acceleration of total metabolism to cause extreme wasting [B. Beutler, D. 
Greenwald, J. D. Hulmes et al., Nature, 316, 552-554 (1985), Kawakami, M., 
SEIKAGAKU (Biochemistry), 59, 1244-1247 (1987)]. 
TNF is one of causes for a septic shock; in an experiment using an 
antibody, its effect has been recognized [Starnes, H. F. Jr., Pearce, M. 
K., Tewari, A., Yim, J. H., Zou, J. C., Abrams, J. S., J. Immunol., 145, 
4185-4191 (1990), Beutler, B., Milsark, I. W., Cerami, A. C., Science, 
229, 869-871 (1985), Hinshaw, L. B., Tekamp-Olson, P., Chang, A. C. K. et 
al., Circ. Shock, 30, 279-292 (1990)]. 
An increased level of TNF is also observed in the synovial fluid or blood 
from patients with rheumatoid arthritis [Tetta, C., Camussi, G., Modena, 
V., Vittorio, C. D., Baglioni, C., Ann. Rheum. Dis., 49, 665-667 (1990)]. 
In addition, there are many other diseases of which a certain role of TNF 
is suspected, e.g., osteoarthritis reported by Venn, G., Nietfeld, J. J., 
Duits, A. J., Brennan, F. M., Arner, E., Covington, M., Billingham, M. E. 
J., Hardingham, T. E., Arthritis Rheum., 36 (6), 819-826 (1993); multiple 
sclerosis reported by Sharief, M. K., Hentges, R., N. Engl. J. Med., 325 
(7), 467-472 (1991); Kawasaki disease reported by Matsubara, T., Furukawa, 
S., Yabuta, K., Clin. Immunol. Immunopathol., 56, 29-36 (1990); 
inflammatory bowel disease such as ulcerative colitis or Crohn's disease 
reported by Murch, S., Walker-Smith, J. A., Arch. Dis. Child, 66, 561 
(1991); Maeda, M., SHOKAKI-TOMENEKI (Digestive Organ and Immunity), 22, 
111-114 (1989), Behcet disease reported by Akoglu, T., Direskeneli, H., 
Yazici, H., Lawrence, R., J. Rheumatol., 17, 1107-1108 (1990); systemic 
lupus erythematosus (SLE) reported by Maury, C. P. J., Teppo, A.-M., 
Arthritis Rheum., 32, 146-150 (1989); graft versus host disease (GvHD) 
reported by Nestel, F. P., Price, K. S., Seemayer, T. A., Lapp, W. S., J. 
Exp. Med., 175, 405-413 (1992); multiple organ failure reported by 
Fujiwara, T., Kawakami, M., RINSHO-I (Clinician), 17 (10), 2006-2008 
(1991); malaria reported by Grau, G. E., Fajardo, L. F., Piguet, P. F. et 
al., Science, 237, 1210-1212 (1987), acquired immune deficiency syndrome 
(AIDS) reported by Kawakami, M., Hayata K., Medical Immunology, 20, 
615-620 (1990), Dezube, B. J., Pardee, A. B., J. Acquir. Immune Defic. 
Syndr., 5, 1099-1104 (1992); meningitis reported by Waage, A., Halstensen, 
A., Espevik, T., Lancet, I, 355-357 (1987); hepatitis reported by Sugano, 
K., KANZO (Liver), 33, 213-218 (1992), Type II diabetes mellitus reported 
by Hotamisligil, G. S., Shargill, N. S., Spiegelman, B. M., Science, 259, 
87-91 (1993), etc. 
From the above publications, it is understood that excessive production of 
TNF sometimes adversely affect the living body. Therefore, further 
investigations are desired to develop TNF inhibitors available for the 
treatment of these diseases. 
Pentoxifylline having a methylxanthine skeleton is known as a compound 
showing an activity of inhibiting TNF. It is reported that this compound 
possesses an activity of preventing death in endotoxin-shocked mice, an 
activity of improving the sense of well-being or preventing a weight loss 
in cancer patients, an activity of preventing experimental allergic 
encephalomyelitis induced on an animal model, and an activity of 
preventing HIV-1 replication, reported by Zabel, P., Schade, F. U., 
Schlaak, M., Immunobiol., 187, 447-463 (1993), Dezube, B. J., Pardee, A. 
B. et al., Cancer Immuno. Immunother., 36, 57-60 (1993), Nataf, S., 
Louboutin, J. P., Chabannes, D., Feve, J. R., Muller, J. Y., Acta Neurol. 
Scand., 38, 97-99 (1993), Fazely, F., Dezube, B. J., Allen-Ryan, J., 
Pardee, A. B., Ruprecht, R. M., Blood, 77, 1653-1656 (1991). In addition, 
glucocorticoid, protease inhibitors, phospholipase A.sub.2 inhibitors, 
lipoxygenase inhibitors, platelet-aggregating factor (PAF) antagonists, 
radical scavengers, prostaglandin F.sub.2 or I.sub.2 and anti-TNF antibody 
are heretofore known as compounds or factors for showing a TNF inhibitory 
activity. 
In the future, the role of TNF in association with diseases will be made 
clearer, using these low molecular compounds or antibodies. However, these 
compounds are accompanied by side effects due to a wide variety of the 
pharmacological activities. Therefore, it is desired to develop highly 
safe compounds based on a novel mechanism. 
As a compound which is one of the effective ingredients of the composition 
of the present invention and has a structure close to the compounds of the 
present invention, there is known a compound named Eschscholtzine or 
crychine. The compound has the following structure: 
##STR1## 
Eschscholtzine is a natural substance isolated from a plant (Manske, R. H. 
F., Shin, K. H., Can. J. Chem., 43 (8), 2180-2182 (1965), Manske, R. H. 
F., Shin, K. H., Battersby, A. R., Shaw, D. F., Can. J. Chem., 43 (8), 
2183-2189 (1965)). This substance is also synthesized by Barker, A. C., 
Battersby, A. R., J. Chem. Soc. (C), 1317-1323 (1967). It is reported that 
the compound has a pharmacological activity as a vasorelaxant (Ko, F.N., 
Wo, Y. C., Lu, S.T., Teng, C. M., J. Pharm. Pharmacol., 45 (8), 707-710 
(1993)). However, no report is found on the activity of inhibiting the 
production or secretion of TNF as contemplated by the present invention. 
SUMMARY OF THE INVENTION 
The present invention provides a pharmaceutical composition for the 
treatment of diseases, based on the activity of inhibiting the production 
or secretion of TNF, in which TNF is considered to take a part, for 
example, in cachexia, septic shock, multiple organ failure, Rhuematoid 
arthritis, inflammatory bowel disease, multiple sclerosis, osteoarthritis 
Behcet disease, systemic lupus erythematosus (SLE), graft versus host 
disease (GvHD), malaria, acquired immune deficiency syndrome (AIDS), 
meningitis, hepatitis, or Type II diabetes mellitus. 
The present inventors have discovered that the compounds represented by 
general formula (1) described below exhibit an activity of inhibiting the 
production or secretion of TNF. The present invention has thus been 
accomplished. 
That is, a first aspect of the present invention relates to a method for 
preventing or treating a disease caused by TNF, which comprises 
administering to a patient a pharmaceutically effective amount of a 
compound represented by general formula (1) below: 
##STR2## 
wherein R.sup.1 represents a hydrogen atom, an alkyl group, an alkenyl 
group, an acyl group or a group shown by formula: 
EQU --X.sup.1 --(CH.sub.2).sub.k R.sup.7 
wherein R.sup.7 represents a halogen atom, a hydroxyl group, an alkoxy 
group, an alkylthio group, a carboxyl group, an alkoxycarbonyl group, an 
aryloxycarbonyl group, a cyano group, an amino group, an alkylamino group, 
a dialkylamino group, a cycloalkyl group, a heterocyclic group, an 
aromatic hydrocarbon group or an aromatic heterocyclic group; X.sup.1 
represents a carbonyl group or a methylene group; k represents 0 or an 
integer of 1 to 5, provided that when X.sup.1 is a carbonyl group and 
R.sup.7 is a hydroxyl group, or when X.sup.1 is a methylene group and 
R.sup.7 is a hydroxyl group, an amino group, an alkylamino group or a 
dialkylamino group, k represents an integer of 1 to 5; 
and, each of A and B independently represents a methylene group or a group 
shown by: 
##STR3## 
wherein each of R.sup.2 and R.sup.3 independently represents a hydrogen 
atom, an alkyl group, an alkoxycarbonyl group or a substituted alkyl 
group; or a pharmaceutically acceptable salt thereof. 
A second aspect of the present invention relates to use of the above 
compound represented by formula (1) for the preparation of a 
pharmaceutical composition for preventing or treating a disease caused by 
TNF. 
A third aspect of the present invention relates to a new compound 
represented by general formula: 
##STR4## 
wherein R.sup.1, A and B are as defined above, provided that when R.sup.1 
is methyl, A and B are not methylene simultaneously; or a salt thereof. 
A fourth aspect of the present invention relates to a pharmaceutical 
composition for inhibiting the preparation or secretion of TNF, which 
comprises as an effective ingredient a pharmaceutically effective amount 
of the above new compound or a pharmaceutically acceptable salt thereof 
and a pharmaceutically acceptable carrier or diluent. 
A fifth aspect of the present invention relates to the above new compound 
for use as a medicament. 
DETAILED DESCRIPTIONS OF THE INVENTION 
The functional groups in the compounds given above are described below in 
more detail. 
In the compounds of the present invention, k represents 0 or an integer of 
1 to 5 when R.sup.1 is shown by the formula: --X.sup.1 --(CH.sub.2).sub.k 
R.sup.7. Compounds of general formula (1) wherein k is 0, 1 or 2 are 
preferred for the present invention. 
As the alkyl group and the alkenyl group, a lower alkyl group and a lower 
alkenyl group are preferred, respectively. As the acyl group, preferred 
are a lower alkanoyl group and an aroyl group having carbon atoms of 11 or 
less, e.g., a benzoyl group. A preferred example of the alkoxy group is a 
lower alkoxy group. As the alkylthio group, the alkoxycarbonyl group, the 
alkylamino group, the dialkylamino group and the cycloalkyl group, 
preferred are a lower alkylthio group, a lower alkoxycarbonyl group, a 
lower alkylamino group, a lower dialkylamino group and a lower cycloalkyl 
group, respectively. 
The lower alkyl group includes a straight or branched alkyl group having 1 
to 6 carbon atoms. Specific examples of the lower alkyl group include 
methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 
2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 
3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 
hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 
1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 
2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 
2-ethylbutyl and 1,1,2-trimethylpropyl. The alkyl groups having 1 to 3 
carbon atoms are preferred. 
The lower alkenyl group includes a straight or branched alkenyl group 
having 2 to 6 carbon atoms. Specific examples of the lower alkenyl group 
include ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 
1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 
3-hexenyl, 4-hexenyl, 5-hexenyl and 3-methyl-2-butenyl. Preferred alkenyl 
group have 2 to 3 carbon atoms. 
The lower alkanoyl group includes a straight or branched alkanoyl group 
having 1 to 6 carbon atoms. Specific examples of the lower alkanoyl group 
include formyl, acetyl, propionyl, butyryl, valeryl, isovaleryl, pivaloyl 
and hexanoyl. 
Examples of the halogen atom include fluorine, chlorine, bromine and 
iodine. 
The lower alkoxy group includes a straight or branched alkoxy group having 
1 to 6 carbon atoms. Specific examples of the lower alkoxy group include 
methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 1-methylpropoxy, 
2-methylpropoxy, pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 
3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 
2,2-dimethylpropoxy, hexyloxy, 1-methylpentyloxy, 2-methylpentyloxy, 
3-methylpentyloxy, 4-methylpentyloxy, 1,1-dimethylbutoxy, 
1,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2,2-dimethylbutoxy, 
2,3-dimethylbutoxy, 3,3-dimethylbutoxy, 1-ethyl-1-methylpropoxy and 
1-ethyl-2-methylpropoxy. 
The lower alkylthio group includes a straight or branched alkylthio group 
having 1 to 6 carbon atoms. Specific examples of the lower alkylthio group 
include methylthio, ethylthio, propylthio, 1-methylethylthio, butylthio, 
1-methylpropylthio, 2-methylpropylthio, pentylthio, 1-methylbutylthio, 
2-methylbutylthio, 3-methylbutylthio, 1,1-dimethylpropylthio, 
1,2-dimethylpropylthio, 2,2-dimethylpropylthio, hexylthio, 
1-methylpentylthio, 2-methylpentylthio, 3-methylpentylthio, 
4-methylpentylthio, 1,1-dimethylbutylthio, 1,2-dimethylbutylthio, 
1,3-dimethylbutylthio, 2,2-dimethylbutylthio, 2,3-dimethylbutylthio, 
3,3-dimethylbutylthio, 1-ethyl-1-methylpropylthio and 
1-ethyl-2-methylpropylthio. 
The lower alkoxycarbonyl group includes a straight or branched 
alkoxycarbonyl group having 2 to 6 carbon atoms. Specific examples of the 
lower alkoxycarbonyl group include methoxycarbonyl, ethoxycarbonyl, 
propoxycarbonyl, 1-methylethoxycarbonyl, butoxycarbonyl and 
1,1-dimethylethoxycarbonyl. 
The aryloxycarbonyl group has preferably 7 to 13 carbon atoms. A specific 
example is phenoxycarbonyl. 
The lower alkylamino group includes an alkylamino group having 1 to 4 
carbon atoms and specific examples include methylamino, ethylamino, 
propylamino, 1-methylethylamino, butylamino and 1,1-dimethylethylamino. 
The di-lower alkylamino group includes a dialkylamino group having 2 to 8 
carbon atoms and specific examples thereof include N,N-dimethylamino, 
N,N-diethylamino, N,N-dipropylamino, N-methyl-N-ethylamino, 
N,N-dibutylamino and N-methyl-N-(1,1-dimethylethyl)amino. 
The lower cycloalkyl group includes a cycloalkyl group having 3 to 7 carbon 
atoms and specific examples include cyclopropyl, cyclobutyl, cyclopentyl 
and cyclohexyl. 
The heterocyclic group includes a monocyclic heterocyclic group which is 
saturated, has carbon atoms of 6 or less and contains as a hetero atom(s) 
one or two nitrogen, oxygen or sulfur atom(s) which may be the same or 
different. More preferably, the monocyclic heterocyclic group is selected 
from 5- and 6-membered heterocyclic groups. Specific examples of the 
5-membered heterocyclic group include 1-pyrrolidinyl, 2-pyrrolidinyl, 
3-pyrrolidinyl, 2-oxolanyl, 3-oxolanyl, 2-thiolanyl and 3-thiolanyl. 
Specific examples of the 6-membered heterocyclic group include piperidino, 
2-piperidyl, 3-piperidyl, 4-piperidyl, 1-piperazinyl, 2-piperazinyl, 
2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, morpholino, 
2-morpholinyl and 3-morpholinyl. 
The monocyclic heterocyclic group may be optionally substituted with an 
alkyl group. 
The aromatic hydrocarbon group has preferably carbon atoms of 10 or less 
and specific examples are phenyl, 1-naphthyl and 2-naphthyl. 
The aromatic hydrocarbon group may be optionally substituted with, e.g., a 
halogen atom, a hydroxyl group, an alkyl group, an alkoxy group, a nitro 
group or a cyano group. 
The aromatic heterocyclic group includes a monocyclic aromatic heterocyclic 
group having carbon atoms of 5 or less which contains as a hetero atom(s), 
which may be the same or different, 1 to 3 nitrogen, oxygen or sulfur 
atoms. Specific examples of the aromatic heterocyclic group include 
2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 
5-pyrimidinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-oxazolyl, 
4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 
3-isothiazolyl, 4-isothiazolyl, 5-isothiazolyl, 2-furyl, 3-furyl, 
2-imidazolyl, 4-imidazolyl, 2-thienyl, 3-thienyl, 2-pyrrolyl, 3-pyrrolyl, 
1-pyrazolyl, 3-pyrazolyl, 4-pyrazolyl, 2-pyrazinyl, 3-pyridazinyl, 
4-pyridazinyl, 1H-1,2,4-triazol-1-yl, 1H-1,2,4-triazol-3-yl, 
1H-1,2,4-triazol-5-yl, 3-oxadiazolyl, 5-oxadiazolyl, 2-thiazolyl, 
4-thiazolyl and 5-thiazolyl. 
As the substituted alkyl group, there are, e.g., a substituted lower alkyl 
group having 1 to 3 carbon atoms of the alkyl moiety, which is substituted 
with, e.g., a hydroxyl group, an amino group, an alkylamino group or a 
dialkylamino group. Herein the alkylamino group and the dialkylamino group 
are preferably a lower alkylamino group and a di-lower alkylamino group, 
respectively. These groups may also be substituted with a hydroxyl group, 
an amino group, an alkylamino group or a dialkylamino group. Specific 
examples of the substituted alkyl group include hydroxymethyl, 
aminomethyl, N-methylaminomethyl, N,N-dimethylaminomethyl, 
N-(2-hydroxyethyl)aminomethyl and 
N-[2-(N,N-dimethylamino)ethyl]aminomethyl. 
Typical examples of the salts which are also covered by the present 
invention include salts with mineral acids such as hydrochloric acid, 
hydrobromic acid, sulfuric acid or phosphoric acid; salts with organic 
carboxylic acids such as formic acid, acetic acid, fumaric acid, maleic 
acid, malic acid, tartaric acid, aspartic acid or glutamic acid; salts 
with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, 
p-toluenesulfonic acid, hydroxybenzenesulfonic acid or 
dihydroxybenzenesulfonic acid; salts with alkali metals such as sodium or 
potassium; salts with alkaline earth metals such as calcium or magnesium; 
salts with organic bases such as trimethylamine, triethylamine or 
pyridine, or ammonium salts. 
The compounds of the present invention include stereoisomers and 
geometrical isomers. The compounds of the present invention also include 
all of the hydrates and crystalline forms. 
The composition of the present invention for inhibiting the production or 
secretion of TNF may be administered orally or parenterally. More 
specifically, the composition may be administered orally in a conventional 
form, e.g., in the form of tablets, capsules, syrup or suspension. The 
composition in a liquid form such as a solution, an emulsion or a 
suspension may be parenterally administered in the form of injection. The 
composition may also be administered rectally in the form of a 
suppository. These pharmaceutical preparations can be prepared in a 
conventional manner by formulating the active ingredient together with a 
conventional carrier, excipient, binder, stabilizer, etc. Where the 
pharmaceutical composition is provided in the form of injection, a 
buffering agent, a dissolution aid, an isotonic agent or the like may also 
be added to the composition. 
Dose of the TNF inhibitor and the time for administration vary depending 
upon conditions, age, body weight and preparation form. In general, the 
daily dose of the TNF inhibitor for adult is in the range of 10 to 500 mg 
for oral administration and, for parenteral administration in the range of 
1 to 100 mg. The composition is administered at the daily dose at once or 
by dividing the daily dose into several times. 
The compounds of the present invention can be synthesized, e.g., by the 
following processes. 
Process 1 
##STR5## 
wherein R.sup.1, A and B are as defined above; and each of X.sup.1 and 
X.sup.2 independently represents a halogen atom such as chlorine, bromine 
or iodine. 
The compound shown by general formula (1) can be prepared by reacting a 
dicatechol compound shown by general formula (3) with a dihalide shown by 
general formula (4) in an inert solvent in the presence of a base and then 
reacting the resulting monocatechol compound represented by general 
formula (5) with a dihalide of general formula (6) in the presence of a 
base. 
The solvent used in the above reaction is typically an aprotic solvent, for 
example, N,N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide 
or acetonitrile. 
As the base, there are inorganic bases such as potassium carbonate, cesium 
carbonate, potassium fluoride, cesium fluoride, sodium hydroxide, and 
sodium hydride. 
In the reaction described above, a proportion of the catechol derivatives 
(3) or (5) to the dihalides (4) or (6) is not particularly limited. In 
general, the dihalides are appropriately used between the equimolar amount 
and the amount more than the equimolar amount, based on the catechol 
derivatives. The reactants are used preferably in an almost equimolar 
amount. The base is used in an amount more than the equimolar amount to 
the dihalides. The reaction is carried out generally at a temperature 
ranging from ice cooling to about 150.degree. C. 
Where A and B represent the same group in general formula (1), the reaction 
is carried out in the presence of a base, using more than 2 mols of the 
dihalide (4), based on the compound of general formula (3). The compound 
of general formula (5) can be led to the compound of general formula (1), 
without isolating the compound (5). 
Process 2 
##STR6## 
wherein A, R.sup.1 and X.sup.1 are as defined above. 
The compound represented by general formula (8) which corresponds to the 
compound of general formula (1), wherein substituent B is ethylenedioxy 
substituted with hydroxymethyl, can be prepared, e.g., by the following 
process. 
The compound shown by general formula (8) can be prepared by reacting the 
monocatechol compound shown by general formula (5) with an epihalohydrin 
shown by general formula (7) in an inert solvent in the presence of a 
base. The solvent, base, reaction temperature and other reaction 
conditions used are similar to those for Process 1 described above. 
Process 3 
##STR7## 
wherein R.sup.1 and A are as defined above; and each of R.sup.4 and 
R.sup.5 independently represents a hydrogen atom, an alkyl group or a 
substituted alkyl group. 
The compound represented by general formula (11) which corresponds to the 
compound of general formula (1), wherein substituent B is an ethylenedioxy 
group having a substituted aminomethyl, can be prepared, e.g., by the 
following process. 
The compound shown by general formula (11) can be prepared by reacting a 
hydroxy compound shown by general formula (8) with methanesulfonyl 
chloride in an inert solvent in the presence of a base and then reacting 
the resulting compound represented by general formula (9) with an amine 
represented by general formula (10). 
Examples of the solvent used in the synthesis of the compound (9) are 
aprotic polar solvents such as N,N-dimethylformamide, dimethylsulfoxide, 
hexamethylphosphoramide, and acetonitrile; hydrocarbons such as benzene, 
toluene or hexane; halogenated hydrocarbons such as dichloromethane, 
chloroform or dichloroethane; and ethers such as tetrahydrofuran, dioxane 
or diethyl ether. 
Examples of the base include organic tertiary amines such as triethylamine, 
pyridine, N,N-dimethylaminopyridine, and N-methylmorpholine. 
In the reaction above, a proportion of methanesulfonyl chloride to the 
hydroxy compound represented by general formula (8) is not particularly 
limited but appropriately chosen between the equimolar amount and the 
amount more than the equimolar amount, based on to the hydroxy compound. 
Preferably, the reactants are used in an almost equimolar amount. 
The base is employed in an amount more than the equimolar amount, based on 
methanesulfonyl chloride. The reaction is carried out at a temperature 
ranging from ice cooling to about room temperature. 
Examples of the solvent, which is used in the synthesis of the compound 
shown by general formula (11) from the compound of general formula (9), 
are aprotic polar solvents such as N,N-dimethylformamide, 
dimethylsulfoxide, hexamethylphosphoramide or acetonitrile; halogenated 
hydrocarbons such as dichloromethane, chloroform, or dichloroethane; and 
ethers such as tetrahydrofuran, dioxane or diethyl ether. 
In the reaction described above, the amine derivative of formula (10) is 
used in an excess amount based on the compound of formula (9). The 
reaction is carried out generally at a temperature ranging from ice 
cooling to about 150.degree. C. 
Processes 1 to 3 are directed to the conversion of the compounds having the 
substituted amino group in the catechol moiety thereof. In addition to 
these processes, the compounds of general formula (1) may also be prepared 
through conversion at the catechol moiety followed by introduction of a 
substituent at the amino group. 
Process 4 
##STR8## 
wherein A and B are as defined above, and R.sup.6 represents an alkyl 
group. 
The compound represented by general formula (13) corresponding to the 
compound of general formula (1), wherein substituent R.sup.1 is a hydrogen 
atom, can be prepared by hydrolysis of an acyl derivative represented by 
general formula (12). 
The hydrolysis may proceed in a solvent mixture of a potassium hydroxide or 
sodium hydroxide aqueous solution with an alcoholic solvent such as 
ethanol, ethylene glycol or methoxyethanol, or with an ether such as 
1,4-dioxane or tetrahydrofuran, at a temperature ranging from room 
temperature to the boiling point of the solvent used. 
Process 5 
##STR9## 
wherein R.sup.1, A, B and X.sup.1 are as defined above. 
The compound represented by general formula (1) can be prepared, e.g., by 
reacting an amino derivative shown by general formula (13) with a halide 
shown by general formula (14) in an inert solvent in the presence of a 
base. 
Examples of the solvent used are aprotic polar solvents such as 
N,N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide or 
acetonitrile; hydrocarbons such as benzene, toluene or hexane; halogenated 
hydrocarbons such as dichloromethane, chloroform or dichloroethane; and 
ethers such as tetrahydrofuran, dioxane or diethyl ether. 
Examples of the base include organic tertiary amines such as triethylamine, 
pyridine, N,N-dimethylaminopyridine or N-methylmorpholine; and inorganic 
bases such as potassium carbonate, sodium carbonate or sodium 
hydrogencarbonate. 
In the reaction above, a proportion of the amino derivative to the halide 
of formula (14) is not particularly limited. In general, the halide (14) 
may be used in an amount more than the equimolar amount based on the amino 
derivative. Preferably, the reactants are used in an almost equimolar 
amount. The base is employed in an amount more than the equimolar amount 
based on the halide. The reaction is carried out at a temperature ranging 
from ice cooling to about the boiling point of the solvent used. 
Process 6 
##STR10## 
wherein A, B, k and R.sup.7 are as defined above. 
The compound represented by general formula (17) corresponding to the 
compound of general formula (1), wherein substituent R.sup.1 is a group 
represented by formula --CO(CH.sub.2).sub.k R.sup.7, can also be prepared 
either by condensing the amine derivative represented by general formula 
(13) with the carboxylic acid represented by general formula (15), or by 
reacting the amine derivative (13) with an acid anhydride shown by general 
formula (16) in the presence of a base, in an inert solvent. 
Examples of the condensing agent used in the condensation are 
N,N'-dicyclohexylcarbodiimide, 
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, etc. The other reaction 
conditions including reaction solvents, bases and reaction temperatures 
are similar to those set forth in Process 5. 
Process 7 
##STR11## 
wherein A, B, R.sup.7 and k are as defined above. 
The compound represented by general formula (19) corresponding to the 
compound of general formula (1), wherein substituent R.sup.1 is a group 
represented by formula --CH.sub.2 (CH.sub.2).sub.k R.sup.7, can be 
prepared, e.g., by reacting the amine derivative shown by general formula 
(13) with an aldehyde shown by general formula (18), in an inert solvent 
in the presence of a reducing agent for reductive amination. 
Examples of the solvent used for the above reaction include alcohol such as 
methanol or ethanol; hydrocarbons such as benzene, toluene or hexane; 
halogenated hydrocarbons such as dichloromethane, chloroform or 
dichloroethane; ethers such as tetrahydrofuran, dioxane or diethyl ether. 
Examples of the reducing agent used for the above reaction include hydride 
compounds such as lithium aluminum hydride, sodium cyanoborohydride, and 
sodium borohydride. These solvent and reducing agent may be used in an 
appropriate combination thereof. 
In the reaction above, a proportion of the amino derivative to the aldehyde 
is not particularly limited. In general, the aldehyde may be used 
appropriately between the equimolar amount and more than the equimolar 
amount, based on the amino derivative. Preferably, the reactants are used 
in an almost equimolar amount. The reaction is carried out at a 
temperature ranging from ice cooling to about the boiling point of the 
solvent used. 
Process 8 
##STR12## 
wherein A, B and R.sup.6 have the same significance as defined above. 
The compound represented by general formula (21) corresponding to the 
compound of general formula (1), wherein substituent R.sup.1 is a group 
represented by formula CONHR.sup.6, can be prepared, e.g., by reacting the 
amino derivative shown by general formula (13) with an isocyanate shown by 
general formula (20) in an inert solvent. 
Examples of the solvent used are aprotic polar solvents such as 
N,N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide or 
acetonitrile; hydrocarbons such as benzene, toluene or hexane; halogenated 
hydrocarbons such as dichloromethane, chloroform or dichloroethane; and 
ethers such as tetrahydrofuran, dioxane or diethyl ether. 
In the reaction above, a proportion of the amino derivative of formula (13) 
to the isocyanate of formula (20) is not particularly limited. In general, 
the isocyanate of formula (20) is used appropriately chosen between the 
equimolar amount and more than the equimolar amount, based on the amino 
derivative. Preferably, the reactants are used in an almost equimolar 
amount. The reaction is carried out at a temperature ranging from ice 
cooling to about the boiling point of the solvent used. 
The dicatechol compound shown by general formula (3) which is one of the 
starting compounds for producing the compounds of the present invention 
may be prepared, e.g., by the following process. 
Process 9 
##STR13## 
wherein R.sup.1 and X.sup.1 are as defined above. 
The dicatechol compound of formula (3) may be prepared by heating 
3,4-dihydroxyphenylserine shown by formula (22) with hydrochloric acid and 
then reacting the resulting amino compound of formula (23) with the halide 
of general formula (14), in a manner similar to Process 5. 
##STR14## 
wherein R.sup.7 and k are as defined above. 
The compound of general formula (24) corresponding to the compound of 
general formula (3), wherein substituent R.sup.1 is a group represented by 
formula --CO(CH.sub.2).sub.k R.sup.7, may be prepared either by condensing 
the compound of formula (23) with the carboxylic acid shown by general 
formula (15) or by reacting the compound (23) with the acid anhydride 
shown by general formula (16), in a manner similar to Process 6. 
##STR15## 
wherein R.sup.7 and k are as defined above. 
The compound of general formula (25) corresponding to the compound of 
general formula (3), wherein substituent R.sup.1 is a group represented by 
formula --CH.sub.2 (CH.sub.2).sub.k R.sup.7, may also be prepared by 
reductive amination of the aldehyde shown by general formula (18), in a 
manner similar to Process 7. 
In the process described above, when R.sup.1, A or B of the compound of 
general formula (1) possess one or more functional group(s) such as an 
amino group, an alkylamino group or a hydroxyl group, such group(s) can be 
protected by protective group(s) before the each step(s) in Process 1 to 
9, and deprotected after the each step(s), if necessary or desired. 
Such a protection-deprotection technique is described in, for example, T. 
W. Greene, "Protective Groups in Organic Synthesis" John Willey & Sons 
Inc., 1981. 
The "protective group" includes following groups; 
1) Protective group for an amino group or an alkylamino group 
an alkanoyl group such as an acetyl group 
an aroyl group such as a benzoyl group 
a tert-butoxycarbonyl group 
a benzyloxycarbonyl group 
a phthaloyl group (only for an amino group) 
2) protective group for a hydroxyl group 
an alkanoyl group such as an acetyl group 
an aroyl group such as a benzoyl group 
a benzyl group 
a methoxymethyl group 
a trimethylsilyl group 
a tetrahydropyranyl group 
a tetrahydrofuranyl group

Hereinafter the present invention will be described in more detail by 
referring to the following examples but is not deemed to be limited 
thereto. 
EXAMPLE 1 
##STR16## 
A mixture of 40 g of 13-acetyl-5, 6, 11, 
12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2, 3, 8, 9-tetrol, 119 g of 
cesium carbonate, 47.4 g of bromochloromethane and 600 ml of 
N,N-dimethylformamide was heated at 100.degree. C. with stirring in a 
nitrogen atmosphere. Three hours after, insoluble salts were filtered off 
and the filtrate was concentrated in vacuo. The residue was partitioned 
between ethyl acetate and water, and the organic phase was dried over 
sodium sulfate. The solvent was distilled off in vacuo. The residue was 
purified by silica gel column chromatography (eluent, 
dichloromethane:ethyl acetate=9:1). The product was dissolved in a small 
quantity of methanol and water was added to the solution for trituration 
to give 28 g of 
15-acetyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5 
,12-imine. Melting point: 144.degree.-145.degree. C. 
The starting 
13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-te 
trol was prepared as follows. 
A mixture of 600 g of 3-(3,4-dihydroxyphenyl)serine and 3.6 liters of 1N 
hydrochloric acid was heated at 90.degree. C. with stirring in a nitrogen 
atmosphere. Five hours after, the reaction solution was cooled and then 
allowed to stand overnight. The formed crystals were filtered. 
The crude product was warmed in a mixture of 600 ml of acetone and 2 liters 
of acetonitrile. Insoluble crystals were filtered to give 275 g of 
5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tetrol 
hydrochloride: 
##STR17## 
Melting point: 225.degree.-229.degree. C. 
Then, 50 g of 
5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tetrol 
hydrochloride and 94.11 g of triethylamine were dissolved in 500 ml of 
N,N-dimethylformamide. Under cooling on an ice bath, 79.12 g of acetic 
anhydride was dropwise added to the solution over 30 minutes in a nitrogen 
atmosphere. The mixture was stirred at room temperature for further 5 
hours. The salt formed was filtered off and the filtrate was concentrated 
in vacuo. The residue was crystallized from ethanol to give 73.15 g of 
2,3,8,9-tetraacetoxy-13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten- 
5,11-imine: 
##STR18## 
Melting point, 236.degree.-237.degree. C. 
A mixture of 73 g of 
2,3,8,9-tetraacetoxy-13-acetyl-5,6,11,12-tetrahydro-dibenzo[a,e]cycloocten 
-5,11-imine, 10 g of potassium carbonate and 1 liter of methanol was then 
heated at 40.degree. C. with stirring in a nitrogen atmosphere. Thirty 
minutes after, the reaction mixture was rendered acidic with acetic acid 
followed by concentration in vacuo. The residue was dissolved in a small 
quantity of methanol and water was added to the solution for trituration. 
The product was filtered to give 48 g of 
13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-te 
trol: 
##STR19## 
Melting point, 294.degree.-296.degree. C. (dec.). 
EXAMPLE 2 
##STR20## 
A mixture of 300 mg of 
13-ethyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tet 
rol hydrochloride, 1.3 g of cesium fluoride, 333 mg of dibromomethane and 6 
ml of N,N-dimethylformamide was heated at 110.degree. C. with stirring in 
a nitrogen atmosphere. An hour and a half later, the reaction mixture was 
concentrated in vacuo. The residue was partitioned between diethyl ether 
and 1N sodium hydroxide and the organic phase was washed with water. After 
the organic phase was dried over sodium sulfate, the solvent was distilled 
off in vacuo. The residue was purified by preparative TLC (developing 
solvent, dichloromethane:methanol=50:1). The product was dissolved in 
ethanol and hydrogen chloride/diethyl ether solution (about 7%) was added 
to the solution. The formed salt was filtered to give 22 mg of 
15-ethyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5, 
12-imine hydrochloride. Melting point: 284.degree.-288.degree. C. (dec.). 
The starting 
13-ethyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tet 
rol hydrochloride was prepared as follows. 
##STR21## 
After 3 g of 
5,6,11,12-tetrahydrodibenzo[a,e]-cycloocten-5,11-imine-2,3,8,9-tetrol 
hydrochloride was dissolved in 28 ml of methanol, 535 mg of acetaldehyde 
and 1.18 g of sodium cyanoborohydride were added to the solution. The 
mixture was stirred at room temperature overnight in a nitrogen 
atmosphere. Furthermore 206 mg of acetaldehyde and 294 mg of sodium 
cyanoborohydride were added to the reaction mixture. After the reaction 
was continued for 6 hours, conc. hydrochloric acid was added to the 
mixture to render the system acidic. 
The solvent was distilled off in vacuo. The residue was crystallized from 
water to give 1.45 g of 
13-ethyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tet 
rol hydrochloride. Melting point, 245.degree.249.degree. C. (dec.). 
EXAMPLE 3 
##STR22## 
In a manner similar to Example 2, 20 mg of 
15-furfuryl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol 
-5,12-imine hydrochloride was obtained from 200 mg of 
13-furfuryl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9- 
tetrol hydrochloride. Melting point, 225.degree.-228.degree. C. (dec.). The 
starting 
13-furfuryl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9- 
tetrol hydrochloride was prepared as follows. 
##STR23## 
After 2.5 g of 
5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-tetrol 
hydrochloride was dissolved in 23 ml of methanol, 1.12 g of furfural and 
979 mg of sodium cyanoborohydride were added to the solution. The mixture 
was stirred at room temperature overnight in a nitrogen atmosphere. 
Furthermore 748 mg of furfural was added to the reaction mixture. After 
the reaction was continued for 8 hours, the reaction mixture was 
concentrated in vacuo. The residue was dissolved in water. While carefully 
adding sodium hydrogencarbonate to the solution, the system was rendered 
basic followed by extraction with ethyl acetate. After washing with water, 
the organic phase was dried over sodium sulfate and the solvent was 
distilled off in vacuo. The residue was dissolved in a small quantity of 
methanol and hydrogen chloride/diethyl ether solution (about 7%) was added 
to the solution to render the system acidic. The salt formed from diethyl 
ether was filtered and recrystallized from methanoldiethyl ether to give 
323 mg of 
13-furfuryl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9- 
tetrol hydrochloride. Melting point, 235.degree.-240.degree. C. (dec.). 
EXAMPLE 4 
##STR24## 
In a manner similar to Example 2, 34 mg of 
15-[3-(4-fluorophenyl)propyl]-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']b 
is[1,3]benzodioxol-5,12-imine hydrochloride was obtained from 350 mg of 
13-[3-(4-fluorophenyl)propyl]-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5 
,11-imine-2,3,8,9-tetrol hydrochloride. Melting point, 
155.degree.-157.degree. C. 
The starting 
13-[3-(4-fluorophenyl)propyl]-5,6,11,12-tetrahydrodibenzo[a,e]cyclooctene- 
5,11-imine-2,3,8,9-tetrol hydrochloride was prepared as follows. 
##STR25## 
After 2.5 g of 
5,6,11,12-tetrahydrodibenzo-[a,e]cycloocten-5,11-imine-2,3,8,9-tetrol 
hydrochloride was dissolved in 23 ml of methanol, 1.54 g of 
3-(4-fluorophenyl)propanal and 979 mg of sodium cyanoborohydride were 
added to the solution. The mixture was stirred at room temperature 
overnight in a nitrogen atmosphere. Furthermore 711 mg of 
3-(4-fluorophenyl)propanal was added to the reaction mixture. After the 
reaction was continued for 8 hours, the reaction mixture was concentrated 
in vacuo. The residue was dissolved in water. While carefully adding 
sodium hydrogen carbonate to the solution, the system was rendered basic 
followed by extraction with ethyl acetate. After washing with water, the 
organic phase was dried over sodium sulfate and the solvent was distilled 
off in vacuo. The residue was dissolved in a small quantity of methanol 
and hydrogen chloride/diethyl ether solution (about 7%) was added to the 
solution to render the system acidic. The salt formed from diethyl ether 
was filtered to give 1.73 g of 
13-[3-(4-fluorophenyl)propyl]-5,6,11,12-tetrahydrodibenzo-[a,e]cycloocten- 
5,11-imine-2,3,8,9-tetrol hydrochloride. Melting point, 
182.degree.-188.degree. C. (dec.). 
EXAMPLE 5 
##STR26## 
A mixture of 10 g of 
13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-te 
trol, 42.8 g of potassium carbonate, 34.9 g of 1,2-dibromoethane and 186 ml 
of N,N-dimethylformamide was heated at 110.degree. C. with stirring in a 
nitrogen atmosphere. Thirty hours after, insoluble salts were filtered off 
and the filtrate was concentrated in vacuo. The residue was partitioned 
between dichloromethane and water, and the organic phase was dried over 
sodium sulfate. The solvent was distilled off in vacuo. The residue was 
purified by silica gel column chromatography (eluent, 
dichloromethane:ethyl acetate=9:1). The product was dissolved in a small 
quantity of methanol and water was added to the solution for trituration 
to give 9.81 g of 
17-acetyl-6,7,14,15-tetrahydrocycloocta[1,2-g:5,6-g']bis[1,4]benzodioxan-6 
,14-imine. Melting point: 170.degree.-175.degree. C. 
EXAMPLE 6 
##STR27## 
A mixture of 353 mg of 
15-acetyl-6,7,12,13-tetrahydrobenzo[5,6]cycloocta[1,2-g]-1,4-benzodioxan-6 
,12-imine-9,10-diol, 415 mg of potassium carbonate, 299 mg of ethyl 
2,3-dibromopropionate and 5 ml of N,N-dimethylformamide was heated at 
100.degree. C. with stirring in a nitrogen atmosphere. Three hours after, 
insoluble salts were filtered off and the filtrate was concentrated in 
vacuo. The residue was partitioned between dichloromethane and water and 
the organic phase was dried over sodium sulfate. The solvent was distilled 
off in vacuo. The residue was purified by silica gel column chromatography 
(eluent, chloroform:methanol=100:1). The product was dissolved in a small 
quantity of methanol and water was added to the solution for trituration 
to give 95 mg of a mixture of 
17-acetyl-2-ethoxycarbonyl-6,7,14,15-tetrahydrocycloocta-[1,2-g:5,6-g']bis 
[1,4]-benzodioxan-6,14-imine and 
17-acetyl-3-ethoxycarbonyl-6,7,14,15-tetrahydrocycloocta[1,2-g:5,6-g']bis[ 
1,4]benzodioxan-6,14-imine. Melting point: 124.degree.-128.degree. C. 
The starting 
15-acetyl-6,7,12,13-tetrahydrobenzo[5,6]cycloocta[1,2-g]-1,4-benzodioxan-6 
,12-imine-9,10-diol was prepared as follows. 
##STR28## 
A mixture of 16 g of 
13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-te 
trol, 7.45 g of potassium carbonate, 10.13 g of 1,2-dibromoethane and 240 
ml of N,N-dimethylformamide was heated at 100.degree. C. with stirring in 
a nitrogen atmosphere. Fifteen hours after, insoluble salts were filtered 
off and the filtrate was concentrated in vacuo. The residue was rendered 
acidic with 1N hydrochloric acid. The insoluble salts were filtered, dried 
and purified by silica gel column chromatography (eluent, 
dichloromethane:methanol=30:1). The product was recrystallized from 
methanol to give 3.91 g of 
15-acetyl-6,7,12,13-tetrahydrobenzo-[5,6]cycloocta[1,2-g]-1,4-benzodioxan- 
6,12-imine-9,10-diol. Melting point: 192.degree.-194.degree. C. 
EXAMPLE 7 
##STR29## 
In a manner similar to Example 6, 303 mg of a mixture of 
17-acetyl-6,7,14,15-tetrahydro-2-hydroxymethylcycloocta[1,2-g:5,6-g']bis[1 
,4]benzodioxan-6,14-imine and 
17-acetyl-6,7,14,15-tetrahydro-3-hydroxymethylcycloocta[1,2-g: 
5,6-g']bis[1,4]benzodioxan-6,14-imine was obtained from 600 mg of 
15-acetyl-6,7,12,13-tetrahydrobenzo[5,6]cycloocta[1,2-g]-1,4-benzodioxan-6 
,12-imine-9,10-diol and 165 mg of epichlorohydrin. Melting point, 
172.degree.-180.degree. C. 
EXAMPLE 8 
##STR30## 
In a manner similar to Example 6, 2.63 g of a mixture of 
16-acetyl-6,7,13,14-tetrahydro-2-hydroxymethyl[1,3]benzodioxolo[5,6-f]cycl 
oocta[1,2-g]-1,4-benzodioxan-6,13-imine and 
16-acetyl-6,7,13,14-tetrahydro-3-hydroxymethyl[1,3]benzodioxolo[5,6-f]cycl 
oocta[1,2-g]-1,4-benzodioxan-6,13-imine was obtained from 4.5 g of 
14-acetyl-5,6,11,12-tetrahydrobenzo[5,6]cycloocta[1,2-f]-1,3-benzodioxol-5 
,11-imine-8,9-diol and 1.29 g of epichlorohydrin. Melting point: 
174.degree.-182.degree. C. 
The starting 
14-acetyl-5,6,11,12-tetrahydrobenzo[5,6]cycloocta[1,2-f]-1,3-benzodioxol-5 
,11-imine-8,9-diol was prepared as follows. 
##STR31## 
A mixture of 38 g of 
13-acetyl-5,6,11,12-tetrahydrodibenzo[a,e]cycloocten-5,11-imine-2,3,8,9-te 
trol, 39.4 g of cesium fluoride, 21.0 g of bromochloromethane and 540 ml of 
N,N-dimethylformamide was heated at 100.degree. C. with stirring in a 
nitrogen atmosphere. Five hours and a half later, insoluble salts were 
filtered off and the filtrate was concentrated in vacuo. The residue was 
rendered acidic with 1N hydrochloric acid. The insoluble solids were 
filtered, dried and purified by silica gel column chromatography 
(eluent:dichloromethane:methanol=30:1). 
The product was recrystallized from methanol to give 8.03 g of 
14-acetyl-5,6,11,12-tetrahydrobenzo-[5,6]cycloocta[1,2-f]-1,3-benzodioxol- 
5,11-imine-8,9-diol. Melting point: 261.degree.-265.degree. C. (dec.). 
EXAMPLE 9 
##STR32## 
A mixture of 300 mg of an isomeric mixture of 
16-acetyl-6,7,13,14-tetrahydro-2-(O-mesylmethyl)-[1,3]benzodioxolo[5,6-f]c 
ycloocta[1,2-g]-1,4-benzodioxan-6,13-imine and 
16-acetyl-6,7,13,14-tetrahydro-3-(O-mesylmethyl)-[1,3]benzodioxolo[5,8-f]c 
ycloocta[1,2-g]-1,4-benzodioxan-6,13-imine, 3 ml of ammonium hydroxide 
(about 29% solution in water) and 3 ml of dioxane were heated at 
80.degree. C. in an autoclave. After heating for 6.5 hours, the reaction 
mixture was concentrated in vacuo and the residue was partitioned between 
dichloromethane and 2N hydrochloric acid. The aqueous phase was rendered 
basic with ammonia water. After extracting again with dichloromethane, the 
organic phase was dried over sodium sulfate. The solvent was distilled off 
in vacuo. The residue was crystallized from diethyl ether to give 158 mg 
of the isomeric mixture of 
16-acetyl-2-aminomethyl-6,7,13,14-tetrahydro-[1,3]benzodioxolo[5,6-f]cyclo 
octa[1,2-g]-1,4-benzodioxan-6,13-imine and 
16-acetyl-3-aminomethyl-6,7,13,14-tetrahydro-[1,3]benzodioxolo[5,6-f]cyclo 
octa[1,2-g]-1,4-benzodioxan-6,13-imine. Melting point: 
180.degree.-187.degree. C. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in tetrahydrofuran. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
235.degree.-241.degree. C. 
The starting methanesulfonyl compound was prepared as follows. 
##STR33## 
After 2 g of an isomeric mixture of 
16-acetyl-6,7,13,14-tetrahydro-2-hydroxymethyl[1,3]benzodioxolo[5,6-f]cycl 
oocta[1,2-g]-1,4-benzodioxan-6,13-imine and 
16-acetyl-6,7,13,14-tetrahydro-3-hydroxymethyl[1,3]benzodioxolo[5,6-f]cycl 
oocta[1,2-g]-1,4-benzodioxan-6,13-imine and 563 mg of triethylamine were 
dissolved in 30 ml of chloroform, 0.43 ml of methanesulfonyl chloride was 
dropwise added to the solution in a nitrogen atmosphere under cooling on 
an ice bath. Stirring was continued at the same temperature for 2 hours 
and then 256 mg of triethylamine and 0.2 ml of methanesulfonyl chloride 
were further added to the mixture. The resulting mixture was then allowed 
to stand overnight in a refrigerator. The reaction mixture was partitioned 
between chloroform and water, and the organic phase was dried over sodium 
sulfate. After the solvent was distilled off in vacuo, the residue was 
purified by silica gel column chromatography (eluent:dichloromethane:ethyl 
acetate=8:2). Thus, 2 g of the isomeric mixture of 
16-acetyl-6,7,13,14-tetrahydro-2-(O-mesylmethyl)-[1,3]benzodioxolo-[5,6-f] 
cycloocta[1,2-g]-1,4-benzodioxan-6,13-imine and 
16-acetyl-6,7,13,14-tetrahydro-3-(O-mesylmethyl)-[1,3]benzodioxolo[5,6-f]c 
ycloocta[1,2-g]-1,4-benzodioxan-6,13-imine was obtained as a foamy 
substance. 
EXAMPLES 10 to 12 
The following compounds were obtained from methanesulfonyl compounds and 
amine derivatives in a manner similar to Example 9. 
##STR34## 
TABLE 1 
______________________________________ 
Example No. 
R.sup.4 R.sup.5 Yield M.P. 
______________________________________ 
10 CH.sub.3 
CH.sub.3 95% 198-205.degree. C. 
hydrochloride 
11 H (CH.sub.2).sub.2 OH 
90% 185-915.degree. C. 
hydrochloride (dec.) 
12 H (CH.sub.2).sub.2 N(CH.sub.3).sub.2 
82% 217-220.degree. C. 
hydrochloride 
______________________________________ 
EXAMPLE 13 
##STR35## 
In a mixture of 250 ml of sodium hydroxide 12N solution in water and 250 ml 
of 2-methoxyethanol, 28 g of 
15-acetyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis-[1,3]benzodioxol- 
5,12-imine was heated under reflux for 12 hours. Thereafter the reaction 
mixture was partitioned between dichloromethane and water. The organic 
layer was then washed with saturated sodium chloride aqueous solution. 
After drying over sodium sulfate, the organic phase was concentrated in 
vacuo. The residue was crystallized from methanol to give 14.2 g of 
5,6,12,13-tetrahydrocycloocta-[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
. Melting point: 201.degree.-202.degree. C. 
This free amino compound was converted into hydrochloride salt by adding a 
hydrogen chloride/diethyl ether solution (about 7%) to a solution of the 
amino compound in diethyl ether. The resulting salt was recrystallized 
from ethanol to obtain the hydrochloride. Melting point:&gt;300.degree. C. 
EXAMPLES 14 to 16 
The following compounds were prepared by hydrolysis in a manner similar to 
Example 13. 
##STR36## 
TABLE 2 
__________________________________________________________________________ 
Example No. 
A B Yield 
M.P. 
__________________________________________________________________________ 
14 Hydro- 
(CH.sub.2).sub.2 -- 
(CH.sub.2).sub.2 -- 90% 266-270.degree. C. 
chloride (dec.) 
15 Hydro- 
(CH.sub.2).sub.2 -- 
--CH.sub.2 CH(CH.sub.2 OH)-- 
44% 237-240.degree. C. 
chloride (dec.) 
16 Hydro- 
(CH.sub.2).sub.2 -- 
--CH.sub.2 CH(CH.sub.2 OH)-- 
70% 245-249.degree. C. 
chloride (dec.) 
__________________________________________________________________________ 
EXAMPLE 17 
##STR37## 
After 309 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 21 mg of triethylamine were dissolved in 5 ml of chloroform, 0.1 ml of 
propionyl chloride was dropwise added to the solution in a nitrogen 
atmosphere under cooling on an ice bath. The mixture was stirred at the 
same temperature for 2 hours. The reaction mixture was then partitioned 
between chloroform and water, and the organic phase was dried over sodium 
sulfate. The solvent was distilled off in vacuo. The residue was purified 
by silica gel column chromatography (eluent, dichloromethane:ethyl 
acetate=25:1). The product was dissolved in a small quantity of methanol 
and water was added to the solution for trituration to give 330 mg of 
5,6,12,13-tetrahydro-15-propionylcycloocta-[1,2-f:5,6-f']bis[1,3]benzodiox 
ol-5,12-imine. Melting point: 202.degree.-203.degree. C. 
EXAMPLES 18-39 
The following compounds were prepared from the amino compounds and various 
halides in a manner similar to Example 17. 
##STR38## 
TABLE 3 
______________________________________ 
Compound Yield Melting 
No. R.sup.1 X.sup.1 
(%) point 
______________________________________ 
18 CO(CH.sub.2).sub.2 CH.sub.3 
Cl 88 193-195.degree. C. 
19 CO(CH.sub.2).sub.4 CH.sub.3 
Cl 74 175-176.degree. C. 
20 COCH.sub.2 OCH.sub.3 
Cl 77 184-186.degree. C. 
21 COCO.sub.2 CH.sub.2 CH.sub.3 
Cl 94 194-195.degree. C. 
22 COCH.sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
Cl 86 197-199.degree. C. 
23 COPh Cl 98 271-273.degree. C. 
24 CO.sub.2 CH.sub.2 CH.sub.3 
Cl 82 201-202.degree. C. 
25 CO.sub.2 (CH.sub.2).sub.4 CH.sub.3 
Cl 83 180-181.degree. C. 
26 CO.sub.2 Ph Cl 79 226-227.degree. C. 
27 *2 CH.sub.3 I 37 256-260.degree. C. 
(dec.) 
28 *2 CH.sub.2 CH.sub.3 
I 88 284-288.degree. C. 
(dec.) 
29 *2 (CH.sub.2).sub.2 CH.sub.3 
I 59 155-156.degree. C. 
30 *2 (CH.sub.2).sub.3 CH.sub.3 
Br 50 270-275.degree. C. 
(dec.) 
31 *2 CH.sub.2 CH(CH.sub.3).sub.2 
Br 78 266-267.degree. C. 
32 *2 CH.sub.2 CHCH.sub.2 
Br 88 207-210.degree. C. 
(dec.) 
33 *2 (CH.sub.2).sub.3 CHCH.sub.2 
Br 85 256-259.degree. C. 
(dec.) 
34 *2 *1 Cl 84 255-256.degree. C. 
(dec.) 
35 *2 (CH.sub.2).sub.2 OH 
Br 99 269-272.degree. C. 
(dec.) 
36 *2 (CH.sub.2).sub.2 CO.sub.2 CH.sub.3 
Br 55 231-232.degree. C. 
(dec.) 
37 *2 CH.sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
Br 68 142-145.degree. C. 
38 *2 (CH.sub.2).sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
Br 45 262-265.degree. C. 
(dec.) 
39 *2 (CH.sub.2).sub.2 CN 
Cl 35 191-195.degree. C. 
(dec.) 
______________________________________ 
*1: group shown below 
##STR39## 
*2: hydrochloride 
EXAMPLE 40 
##STR40## 
After 309 mg of 
5,6,12,13-tetrahydrocycloocta-[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
, 55 mg of formic acid and 297 mg of 4-dimethylaminopyridine were dissolved 
in 20 ml of dimethylformamide, 466 mg of 
1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride was added to 
the solution in a nitrogen atmosphere under cooling on an ice bath. After 
reverting to room temperature, the mixture was stirred overnight. The 
reaction mixture was concentrated in vacuo. The residue was then 
partitioned between dichloromethane and 1N hydrochloric acid, followed by 
washing with water. The organic phase was dried over sodium sulfate. The 
solvent was distilled off in vacuo and the residue was purified by silica 
gel column chromatography (eluent, dichloromethane:ethyl acetate=19:1). 
The product was dissolved in a small quantity of methanol and water was 
added to the solution for trituration to give 285 mg of 
15-formyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5 
,12-imine- Melting point: 141.degree.-146.degree. C. 
EXAMPLE 41 
##STR41## 
After 1.3 g of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3S]benzodioxol-5,12-imine 
and 510 mg of triethylamine were dissolved in 21 ml of chloroform, 472 mg 
of acetic anhydride was dropwise added to the solution in a nitrogen 
atmosphere under cooling on an ice bath. At the same temperature, the 
mixture was stirred for 2 hours. The reaction mixture was then partitioned 
between chloroform and water and the organic phase was dried over sodium 
sulfate. The solvent was distilled off in vacuo and the residue was 
purified by silica gel column chromatography (eluent, 
dichloromethane:ethyl acetate=9:1). The product was dissolved in a small 
quantity of methanol and water was added to the solution for trituration 
to give 1.3 g of 
15-acetyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5 
,12-imine. Melting point: 144.degree.-145.degree. C. 
EXAMPLE 42 
##STR42## 
After 346 mg of 
5,6,12,13-tetrahydrocycloocta-[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
hydrochloride and 192 mg of methoxyacetaldehyde (70% aqueous solution) 
were dissolved in methanol, 165 mg of sodium cyanoborohydride was added to 
the solution at room temperature. The mixture was stirred at the same 
temperature overnight. The reaction mixture was then concentrated in 
vacuo. The residue was partitioned between ethyl acetate and water and the 
organic phase was dried over sodium sulfate. The solvent was distilled off 
in vacuo. The residue was purified by silica gel column chromatography 
(eluent, dichloromethane:ethyl acetate=9:1). The product was 
recrystallized from methanol to give 330 mg of 
5,6,12,13-tetrahydro-15-(2-methoxyethyl)cycloocta[1,2-f:5,6-f']bis[1,3]ben 
zodioxol-5,12-imine. Melting point: 187.degree.-188.degree. C. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in dichloromethane. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
240.degree.-244.degree. C. (dec.). 
EXAMPLE 43 
##STR43## 
In a manner similar to Example 42, 356 mg of 
15-[3-(4-fluorophenyl)propyl]-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']b 
is[1,3]benzodioxol-5,12-imine hydrochloride was obtained from 346 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 228 mg of 3-(4-fluorophenyl)propanal- Melting point: 
147.degree.-148.degree. C. 
EXAMPLE 44 
##STR44## 
After 309 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
was dissolved in 5 ml of chloroform, 0.06 ml of methyl isocyanate was 
dropwise added to the solution in a nitrogen atmosphere under cooling on 
an ice bath. Two hours after, the precipitates formed were filtered and 
recrystallized from methanol to give 316 mg of 
5,6,12,13-tetrahydro-15-(N-methylcarbamoyl)cycloocta[1,2-f:5,6-f']bis[1,3] 
benzodioxol-5,12-imine. Melting point: 254.degree.-256.degree. C. 
EXAMPLE 45 
##STR45## 
In a mixture 5 ml of sodium hydroxide 1N solution in water, 10 ml of water 
and 10 ml of tetrahydrofuran was stirred 1 g of 
15-(ethoxycarbonylacetyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-g']bis[1 
,3]benzodioxol-5,12-imine obtained in Example 22. An hour after, the 
reaction solution was partitioned between ethyl acetate and 4N 
hydrochloric acid. The extract was further washed with saturated sodium 
chloride aqueous solution. After the organic phase was dried over sodium 
sulfate, the solvent was distilled off in vacuo. 
The residue was dissolved in a small quantity of methanol and water was 
added to the solution for trituration to give 820 mg of 
15-(carboxyacetyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benz 
odioxol-5,12-imine. Melting point: 154.degree.-159.degree. C. 
EXAMPLE 46 
##STR46## 
In a manner similar to Example 45, 295 mg of 
5,6,12,13-tetrahydro-15-oxalocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5, 
12-imine was obtained from 320 mg of 
15-ethoxalyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxo 
l-5,12-imine obtained in Example 21. Melting point: 183.degree.-187.degree. 
C. 
EXAMPLE 47 
##STR47## 
A solution of 4 g of 
15-(ethoxycarbonylacetyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1 
,3]benzodioxol-5,t2-imine obtained in Example 22 in 50 ml of 
tetrahydrofuran was dropwise added over 30 minutes to a suspension of 206 
mg of lithium borohydride in 50 ml of tetrahydrofuran, at room temperature 
in a nitrogen atmosphere. Stirring was then continued for further 5 hours. 
Then 1N hydrochloric acid was added to the reaction mixture to decompose 
an excess of lithium borohydride. The resulting mixture was concentrated 
in vacuo. 
The residue was partitioned between dichloromethane and water and the 
organic phase was washed with saturated sodium chloride aqueous solution. 
After the organic phase was dried over sodium sulfate, the solvent was 
distilled off in vacuo. The residue was purified by silica gel column 
chromatography (eluent, dichloromethane:ethyl acetate=9:1) to give as a 
foamy substance 1.32 g of 
5,6,12,13-tetrahydro-15-(3-hydroxypropionyl)cycloocta[1,2-f:5,6-f']bis[1,3 
]benzodioxol-5,12-imine. 
EXAMPLE 48 
##STR48## 
In a manner similar to Example 47, 194 mg of 
15-glycoloyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis-[1,3]benzodiox 
ol-5,12-imine was obtained from 409 mg of 
15-ethoxalyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxo 
l-5,12-imine obtained in Example 21. Melting point: 184.degree.-187.degree. 
C. 
EXAMPLE 49 
##STR49## 
A solution of 300 mg of 
15-(2-cyanoethyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzo 
dioxol-5,12-imine in 5 ml of tetrahydrofuran was dropwise added to a 
suspension of 31 mg of lithium aluminum hydride in 1 ml of tetrahydrofuran 
over 20 minutes at room temperature in a nitrogen atmosphere. Stirring was 
then continued for further 5 hours. Then 1N sodium hydroxide was added to 
the reaction mixture to decompose an excess of lithium aluminum hydride. 
The mixture was filtered through celite. The filtrate was concentrated in 
vacuo. The residue was purified by silica gel column chromatography 
(eluent, methanol:ammonium hydroxide (29% aqueous solution)=50:1) to give 
as a foamy substance 50 mg of 
15-(3-aminopropyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,S]benz 
odioxol-5,12-imine. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in tetrahydrofuran. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
215.degree.-220.degree. C. (dec.). 
EXAMPLE 50 
##STR50## 
A mixture of 430 mg of 
5,6,12,13-tetrahydro-15-(2-phthalimidoethyl)cycloocta[1,2-f:5,6-f']bis[1,3 
]benzodioxol-5,12-imine and 0.09 ml of hydrazine monohydrate was stirred in 
6 ml of ethanol at room temperature. Eight hours after, the reaction 
mixture was concentrated in vacuo and the residue was partitioned between 
dichloromethane and saturated sodium bicarbonate aqueous solution. After 
the organic phase was extracted with 2N hydrochloric acid, the aqueous 
phase was rendered basic with sodium hydroxide 2N solution in water and 
extracted again with dichloromethane. After washing with water, the 
organic phase was dried over sodium sulfate. The solvent was distilled off 
in vacuo to give as a foamy substance 183 mg of 
15-(2-aminoethyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzo 
dioxol-5,12-imine. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in dichloromethane. The resulting salt was recrystallized from 
ethanol to obtain the hydrochloride. Melting point: 
235.degree.-240.degree. C. (dec.). 
The starting 
5,6,12,13-tetrahydro-15-(2-phthalimidoethyl)cycloocta[1,2-f:5,6-f']bis[1,S 
]benzodioxol-5,12-imine: 
##STR51## 
was obtained as a foamy substance from 450 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 424 mg of N-(2-bromoethyl)phthalimide in a manner similar to Example 
17. 
EXAMPLE 51 
##STR52## 
In a manner similar to Example 50, 210 mg of 
15-(4-aminobutyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzo 
dioxol-5,12-imine was obtained as a foamy substance from 400 mg of 
5,6,12,13-tetrahydro-15-(4-phthalimidobutyl)cycloocta[1,2-f:5,6-f']bis[1,3 
]benzodioxol-5,12-imine. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in dichloromethane. The resulting salt was recrystallized from 
isopropyl alcohol-ethanol to obtain the hydrochloride. Melting point: 
239.degree.-243.degree. C. (dec.). 
The starting 
5,6,12,13-tetrahydro-15-(4-phthalimidobutyl)cycloocta[1,2-f:5,6-f']bis[1,3 
]benzodioxol-5,12-imine: 
##STR53## 
was obtained from 450 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and N-(4-bromobutyl)phthalimide in a manner similar to Example 17, 
followed by recrystallization of the crude product from ethanol. The yield 
was 569 mg. Melting point: 151.degree.-153.degree. C. 
EXAMPLE 52 
##STR54## 
After 501 mg Of 
15-(N-benzyloxycarbonylglycyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f'] 
bis[1,3]benzodioxol-5,12-imine was dissolved in a solvent mixture of 10 ml 
of methanol and 10 ml of tetrahydrofuran, 100 mg of 10% wet Pd/C was added 
to the solution. Hydrogenation was performed at room temperature under 
normal pressure. Four hours after, the reaction solution was filtered 
through celite and the solvent was distilled off in vacuo. 
The residue was dissolved in a small quantity of methanol and water was 
added to the solution for trituration to give 341 mg of 
15-glycyl-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5 
,12-imine. Melting point: 150.degree.-153.degree. C. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in dichloromethane. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
243.degree.-248.degree. C. (dec.). 
The starting 
15-(N-benzyloxycarbonylglycyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f'] 
bis[1,3]benzodioxol-5,12-imine: 
##STR55## 
was obtained from 500 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 373 mg of benzyloxycarbonylglycine in a manner similar to Example 40. 
The yield was 664 mg. Melting point: 180.degree.-181.degree. C. 
EXAMPLE 53 
##STR56## 
After 560 mg of 
15-(3-N-t-butoxycarbonylaminopropionyl)-5,6,12,13-tetrahydrocycloocta[1,2- 
f:5,6-f']bis[1,3]benzodioxol-5,12-imine was dissolved in 5 ml of 
dichloromethane, 0.3 ml of trifluoroacetic acid was dropwise added to the 
solution in a nitrogen atmosphere under cooling on an ice bath. 
Four hours after, the reaction mixture was partitioned between 
dichloromethane and 1N sodium hydroxide aqueous solution. The organic 
phase was washed with saturated sodium chloride aqueous solution. After 
the organic phase was dried over sodium sulfate, the solvent was distilled 
off in vacuo. The residue was crystallized from diethyl ether to give 412 
mg of 
15-(3-aminopropionyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]b 
enzodioxol-5,12-imine. Melting point: 147.degree.-154.degree. C. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in tetrahydrofuran. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
222.degree.-225.degree. C. 
The starting 
15-(3-N-t-butoxycarbonyl-aminopropionyl)-5,6,12,13-tetrahydrocycloocta[1,2 
-f:5,6-f']bis[1,3]benzodioxol-5,12-imine: 
##STR57## 
was obtained as a foamy substance from 620 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 454 mg of N-t-butoxycarbonyl-.beta.-alanine in a manner similar to 
Example 40. The yield was 670 mg. 
EXAMPLE 54 
##STR58## 
In a manner similar to Example 53, 431 mg of 
15-(4-aminobutyryl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]ben 
zodioxol-5,12-imine was obtained from 560 mg of 
15-(4-N-t-butoxycarbonyl-aminobutyryl)-5,6,12,13-tetrahydrocycloocta[1,2-f 
:5,6-f']bis[1,3]benzodioxol-5,12-imine. Melting point: 
148.degree.-152.degree. C. 
This free amino compound was rendered acidic by adding a hydrogen 
chloride/diethyl ether solution (about 7%) to a solution of the amino 
compound in tetrahydrofuran. The resulting salt was thoroughly washed with 
diethyl ether to obtain the hydrochloride. Melting point: 
202.degree.-205.degree. C. 
The starting 
15-(4-N-t-butoxycarbonylaminobutyl)-5,6,12,13-tetrahydrocycloocta[1,2-f:5, 
6-f']bis[1,3]benzodioxol-5,12-imine: 
##STR59## 
was obtained as a foamy substance from 620 mg of 
5,6,12,13-tetrahydrocycloocta[1,2-f:5,6-f']bis[1,3]benzodioxol-5,12-imine 
and 488 mg of N-t-butoxycarbonyl-4-aminobutyric acid in a manner similar 
to Example 40. The yield was 620 mg. 
EXAMPLE 55 
Inhibitory effect on TNF production or secretion in mouse peritoneal 
macrophages (In case drug concentration is 30 .mu.M) 
BALB/c mice (5 weeks old, female, Charles River Japan) were 
intraperitoneally injected with 1 ml of 3% thioglycollate broth. After 4 
days, the mice were sacrificed. Peritoneal exudated cells (PECs) were 
collected from the peritoneal cavity by washing with minimum essential 
medium (hereinafter abbreviated as MEM, manufactured by Handai 
Biseibutubyo Kenkyukai, Osaka, Japan) containing 5 U/ml heparin and 1% 
fetal bovine serum (FBS, manufactured by GIBCO Laboratories Inc.). PECs 
were washed three times with MEM, suspended with MEM containing 10% FBS. 
After the viable cells were counted by exclusion of trypan blue dye, the 
suspension was adjusted at the final concentration of 2.times.10.sup.6 
cells/ml with MEM containing 10% FBS and seeded into a 96-well microplate 
(Costar, Cambridge, Mass., USA) at 2.times.10.sup.5 cells/100 .mu.l/well. 
The PECs were incubated for an hour at 37.degree. C. in a humidified 5% 
CO.sub.2 incubator, and were washed twice with MEM warmed at 37.degree. C. 
to remove non-adherent cells. Residual adherent cells were used as 
peritoneal macrophages. After the washing above, 50 .mu.l each/well of MEM 
containing 10% FBS was added to each well and provided for use in the 
following experiment. 
The powdery compound of the present invention was dissolved in 
dimethylsulfoxide in a concentration of 30 mM. The solution was then 
diluted with MEM containing 10% FBS in the final concentration of 30 
.mu.M. In the peritoneal macrophages obtained above, 50 .mu.l each of the 
dilution was added to each well to make the total volume 100 .mu.l. 
Thereafter 100 .mu.l each of lipopolysaccharide (hereinafter abbreviated 
as LPS, E. coli 0111B4, manufactured by DIFCO, USA) was added to each well 
in the final concentration of 10 .mu.g/ml. After the cells were incubated 
at 37.degree. C. for 18 hours in a humidified 5% CO.sub.2 incubator, 25 
.mu.l of the supernatant in each well was collected. 
The TNF activity in the supernatant collected was determined by bioassay 
using TNF-sensitive mouse fibroblast cell line L929 cells. That is, 100 
.mu.l each of MEM containing 10% FBS was added to each well of a 96-well 
microplate; using the resulting mixture, 25 .mu.l of the collected 
supernatant was diluted to 5-fold dilution to final concentrations 
(concentrations after the following addition of L929 cell suspension) of 
10%, 2%, 0.4% and 0.08%. 
L929 cells were then suspended (4.times.10.sup.5 cells/ml) in MEM 
containing 10% FBS and 1 .mu.g/ml actinomycin D (Sigma Co.) and 100 .mu.l 
each of the suspension was added to each well of the above microplate at 
4.times.10.sup.4 cells/well and cultured at 37.degree. C. in a humidified 
50% CO.sub.2. The viable cells were counted by partial modification of the 
MTT method reported in Monosann et al., T., J. Immunol. Method, 65, 55-63, 
1983. The modified MTT method comprises the following steps. One mg/ml of 
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (hereinafter 
abbreviated as MTT, manufactured by Sigma Co.) was dissolved in MEM, and 
50 .mu.l each of the solution was added to each well of the microplate 
above. After incubating the microplate for further 6 hours, the 
supernatant was discarded and 100 .mu.l of 0.004N HCl-isopropyl alcohol 
and then 10 .mu.l of 0.01% sodium laurylsulfate aqueous solution was added 
to each well. After shaking the 96-well microplate for a few minutes, the 
absorbance in each well was measured with a microplate reader (Corona Co.) 
at an absorption wavelength of 550 nm. The absorbance correlated to the 
count of the viable L929 cells and represented the TNF activity in the 
supernatant. The TNF activity was determined in terms of unit (U)/ml from 
the calibration curve of absorbance for the TNF activity obtained using 
mouse recombinant TNF .alpha. (TNF-M, manufactured by Genzyme Co.) as a 
standard. The activity of inhibiting TNF production of each compound, was 
determined by the following equation. 
EQU Inhibition of TNF production or secretion (%)=(1-TNF activity in the 
supernatant of the treated cells/TNF activity in the supernatant of the 
non-treated cells).times.100 
The results are shown in Tables 4 through 6. 
##STR60## 
TABLE 4 
__________________________________________________________________________ 
Inhibition of TNF Production or Secretion 
Example Inhibition 
No. R.sup.1 A B (%) 
__________________________________________________________________________ 
1 COCH.sub.3 CH.sub.2 
CH.sub.2 95 
2 CH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 72 
Hydro- 
chloride 
3 Hydro- chloride 
CH.sub.2 
CH.sub.2 -99 
4 Hydro- chloride 
##STR61## CH.sub.2 
CH.sub.2 18 
5 COCH.sub.3 (CH.sub.2).sub.2 
(CH.sub.2).sub.2 64 
6 COCH.sub.3 (CH.sub.2).sub.2 
CH.sub.2 CH(CO.sub.2 CH.sub.2 CH.sub.3) 
89 
7 COCH.sub.3 (CH.sub.2).sub.2 
CH.sub.2 CH(CH.sub.2 OH) 
65 
8 COCH.sub.3 CH.sub.2 
CH.sub.2 CH(CH.sub.2 OH) 
86 
9 COCH.sub.3 CH.sub.2 
CH.sub.2 CH(CH.sub.2 NH.sub.2) 
69 
Hydro- 
chloride 
10 COCH.sub.3 CH.sub.2 
CH.sub.2 CH(CH.sub.2 N(CH.sub.3).sub.2) 
69 
Hydrochloride 
11 COCH.sub.3 CH.sub.2 
CH.sub.2 CH(CH.sub.2 NH(CH.sub.2).sub.2 
61) 
Hydrochloride 
12 COCH.sub.3 CH.sub.2 
CH.sub.2 CH(CH.sub.2 NH(CH.sub.2).sub.2 
N(CH.sub.3).sub.2) 
57 
Hydrochloride 
13 H CH.sub.2 
CH.sub.2 78 
Hydrochloride 
14 H (CH.sub.2).sub.2 
(CH.sub.2).sub.2 63 
Hydrochloride 
15 H CH.sub.2 
CH.sub.2 CH(CH.sub.2 OH) 
55 
Hydrochloride 
16 H (CH.sub.2).sub.2 
CH.sub.2 CH(CH.sub.2 OH) 
18 
Hydrochloride 
__________________________________________________________________________ 
TABLE 5 
__________________________________________________________________________ 
Inhibition of TNF Production or Secretion 
Example Inhibition 
No. R.sup.1 A B (%) 
__________________________________________________________________________ 
17 COCH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
94 
18 CO(CH.sub.2).sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
86 
19 CO(CH.sub.2).sub.4 CH.sub.3 
CH.sub.2 
CH.sub.2 
16 
20 COCH.sub.2 OCH.sub.3 
CH.sub.2 
CH.sub.2 
92 
21 COCO.sub.2 CH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
69 
22 COCH.sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
77 
23 COPh CH.sub.2 
CH.sub.2 
39 
24 CO.sub.2 CH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
68 
25 CO.sub.2 (CH.sub.2).sub.4 CH.sub.3 
CH.sub.2 
CH.sub.2 
46 
26 CO.sub.2 Ph CH.sub.2 
CH.sub.2 
27 
27 CH.sub.3 CH.sub.2 
CH.sub.2 
86 
Hydrochloride 
29 (CH.sub.2).sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
31 
Hydrochloride 
30 (CH.sub.2).sub.3 CH.sub.3 
CH.sub.2 
CH.sub.2 
-37 
Hydrochloride 
31 CH.sub.2 CH(CH.sub.3).sub.2 
CH.sub.2 
CH.sub.2 
36 
Hydrochloride 
32 CH.sub.2 CHCH.sub.2 
CH.sub.2 
CH.sub.2 
12 
Hydrochloride 
33 (CH.sub.2).sub.3 CHCH.sub.2 
CH.sub.2 
CH.sub.2 
-5 
Hydrochloride 
34 Hydrochloride 
##STR62## CH.sub.2 
CH.sub.2 
56 
35 (CH.sub.2).sub.2 OH 
CH.sub.2 
CH.sub.2 
65 
Hydrochloride 
36 (CH.sub.2).sub.2 CO.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
-24 
Hydrochloride 
37 CH.sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
CH.sub.2 
CH.sub.2 
19 
Hydrochloride 
__________________________________________________________________________ 
TABLE 6 
______________________________________ 
Inhibition of TNF Production or Secretion 
Example Inhibition 
No. R.sup.1 A B (%) 
______________________________________ 
38 
(CH.sub.2).sub.2 CO.sub.2 CH.sub.2 CH.sub.3 
--CH.sub.2 -- 
--CH.sub.2 -- 
6 
Hydro- 
chloride 
39 
(CH.sub.2).sub.2 CN 
--CH.sub.2 -- --CH.sub.2 -- 
-48 
Hydro- 
chloride 
40 CHO --CH.sub.2 -- 
--CH.sub.2 -- 
84 
42 
(CH.sub.2).sub.2 OCH.sub.3 
--CH.sub.2 -- --CH.sub.2 -- 
11 
44 --CONHCH.sub.3 --CH.sub.2 -- 
--CH.sub.2 -- 
90 
45 --COCH.sub.2 CO.sub.2 H 
--CH.sub.2 -- 
--CH.sub.2 -- 
15 
46 --COCO.sub.2 H --CH.sub.2 -- 
--CH.sub.2 -- 
6 
47 --CO(CH.sub.2).sub.2 OH 
--CH.sub.2 -- 
--CH.sub.2 -- 
83 
48 --COCH.sub.2 OH 
--CH.sub.2 -- 
--CH.sub.2 -- 
85 
49 
(CH.sub.2).sub.3 NH.sub.2 
--CH.sub.2 -- --CH.sub.2 -- 
-4 
Hydro- 
chloride 
50 
(CH.sub.2).sub.2 NH.sub.2 
--CH.sub.2 -- --CH.sub.2 -- 
64 
Hydro- 
chloride 
51 
(CH.sub.2).sub.4 NH.sub.2 
--CH.sub.2 -- --CH.sub.2 -- 
-44 
Hydro- 
chloride 
52 --COCH.sub.2 NH.sub.2 
--CH.sub.2 -- 
--CH.sub.2 -- 
20 
Hydro- 
chloride 
53 --CO(CH.sub.2).sub.2 NH.sub.2 
--CH.sub.2 -- 
--CH.sub.2 -- 
-1 
Hydro- 
chloride 
54 --CO(CH.sub.2).sub.3 NH.sub.2 
--CH.sub.2 -- 
--CH.sub.2 -- 
35 
Hydro- 
chloride 
______________________________________ 
EXAMPLE 56 
Inhibitory effect on TNF production in mouse peritoneal macrophages (In 
case drug concentrations is 50 .mu.M and 100 .mu.M) 
The inhibitory effect of the inhibitors of the present invention against 
TNF production or secretion was examined at the final concentration of 50 
.mu.M or 100 .mu.M in a manner similar to Example 55. 
The results are shown in Table 7. 
TABLE 7 
______________________________________ 
Inhibition of TNF Production 
or secretion at 50 .mu.M or 100 .mu.M 
Concentration of 
Example No. compound (.mu.M) 
Inhibition (%) 
______________________________________ 
19 100 51 
29 100 81 
Hydrochloride 
30 50 41 
Hydrochloride 
30 100 78 
Hydrochloride 
49 50 21 
Hydrochloride 
______________________________________ 
The results of Table 7 reveal that by increasing the concentration of the 
compound, the inhibition can be enhanced even with the compounds having a 
small or negative value of the TNF inhibitory activity in Example 55. 
EXAMPLE 57 
Protective effect on endotoxin-induced death in galactosamine-treated mice 
It is known that administration of LPS to mice induces a typical shock to 
cause sudden death of the animals. This model is thus considered to be an 
endotoxin-induced shock model. 
It is also suggested that TNF would act as a major mediator for development 
of the disease in this model because a temporarily increased level of TNF 
in mouse blood is observed immediately after the administration of LPS, 
and death due to the shock is prevented by the administration of anti-TNF 
antibody [J. Immunol., 148, 1890-1897 (1992), Lymphokine and Cytokine 
Res., 10 (2), 127-131 (1991), and Science, 229 867-871 (1985)]. 
On the other hand, it is reported that the administration of galactosamine 
results in markedly increased sensitivity to LPS-induced shock so that 
galactosamine is often administered in combination with LPS in the 
endotoxin-induced shock model [Proc. Natl. Acad. Sci. USA, 76 (11), 
5939-5943 (1979), Infect. Immun., 59 (6), 2110-2115 (1991), and J. Infect. 
Dis., 165, 501-505 (1992)]. 
In order to demonstrate the usefulness of the TNF inhibitors of the present 
invention in the endotoxin-induced shock, the following test was performed 
using the evaluation system described above. 
Method 
D-Galactosamine hydrochloride (hereinbelow abbreviated as D-galN, 
manufactured by Nakarai Tesque) and LPS were dissolved in water at final 
concentrations of 75 mg/ml and 0.2 .mu.g/ml, respectively. Furthermore, 
the compound of the present invention was dissolved in a 5% 
dimethylsulfoxide-10% Nikkol (Nippon Surfactant Kogyo, Japan) solution at 
the final concentration of 5 mg/ml. 
BALB/c mice (female, 5 weeks old) obtained from Charles River Japan, Inc. 
were injected i.v. with the aqueous solution containing D-galN and LPS 
described above in a dose of 200 .mu.l/20 g body weight. The mice were 
divided groups of 10 mice. Immediately after the i.v. injection, the 
animals received i.p. injection with the compound of the present invention 
dissolved in the 5% dimethylsulfoxide-10% Nikkol solution in the 
concentration above in a dose of 200 .mu.l/20 g body weight. Control 
animals received the same volume of the 5% dimethylsulfoxide-10% Nikkol 
aqueous solution alone. 
The activity of the compound for protection of endotoxin-induced death in 
the galactosamine-treated mice is expressed in terms of the survival rate 
observed for the following last 7 days. The surviva rate expressed as the 
live/total ratio were statistically analyzed by X.sup.2 method between the 
treated group and the control group. 
Table 7 shows the activity of the respective compounds in the respective 
doses for protection of endotoxin-induced death in the 
galactosamine-treated mice. 
TABLE 7 
______________________________________ 
Protective effect on endotoxin-induced death in 
galactosamine-treated mice 
Compound No. Dose Survival rate 
(Example No.) (mg/kg) (%) 
______________________________________ 
1 0 30 
50 90* 
2 0 0 
Hydrochloride 50 90* 
5 0 20 
50 30 
13 0 10 
Hydrochloride 50 95* 
14 0 20 
Hydrochloride 50 30 
17 0 20 
50 50 
20 0 0 
50 0 
21 0 20 
50 40 
24 0 20 
50 60 
25 0 20 
50 30 
34 0 0 
Hydrochloride 50 70* 
35 0 0 
Hydrochloride 50 90* 
40 0 20 
50 60 
50 0 20 
Hydrochloride 50 100* 
______________________________________ 
*P&lt;0.01 
Preparation Example 1 
Tablet is prepared, e.g., by the following procedure. 
______________________________________ 
mg/tablet 
______________________________________ 
1 Compound of Example 50, hydrochloride 
10 
2 Lactose 72.5 
3 Corn starch 30 
4 Carboxymethyl cellulose Calcium 
5 
5 Hydroxypropyl cellulose (HPC-L) 
2 
6 Magnesium stearate 0.5 
Total 120 mg 
______________________________________ 
The components 1-4 are mixed, agglomerated with aqueous solution of 
component 5, and then mixed with component 6. The resulting mixture is 
compacted into a tablet of 120 mg. 
Preparation Example 2 
Injection is prepared, e.g., by the following procedure. 
Compound of Example 50, hydrochloride 10 mg/vial 
Saline 10 ml/vial 
A solution of the above components is sterilized by filteration, filled in 
a vial previously washed and sterilized. The vial is plugged with a rubber 
stopper washed and sterilized, and then sealed with a flip-off-cap to 
prepare an injection.