AU713107B2 - New pyrido-thiopyranoindazoles with antitumor activity - Google Patents

Description

WO 97/02267 PCT/US96/10966 NEW PYRIDO-THIOPYRANOINDAZOLES WITH ANTI-TUMOR

ACTIVITY

BACKGROUND OF THE INVENTION Field of the invention This invention is directed to 2H-pyrido[3',2':5,6]thiopyrano[4,3,2-cd]indazoles, 2H-pyrido[4',3':5,6]thiopyrano[4,3,2-cd]indazoles, 2Hpyrido[3'4':5,6]thiopyrano[4,3,2-cd]indazoles and 2Hpyrido[2',3':5,6]thiopyrano[4,3,2-cd]indazoles substituted in the positions 2 and o1 These compounds have been shown to have antitumor activity.

Background Certain 1, 4 -bis[(aminoalkyl)amino]anthracene-9,10-diones have been reported which show antitumor activity in clinical trials. Of particular interest has been ametantrone, 1,4-bis[(2-(2-hydroxyethylamino)ethyl)amino]anthracene-9,10dione and mitoxantrone, 5,8-dihydroxy-1,4-bis[(2-(2hydroxyethylamino)ethyl)amino]anthracene-9,10-dione [Zee-Cheng et al., J.

Med. Chem., 21, 291-4 (1978); Cheng et al., "Progress in Medicinal Chemistry", Ellis, G.P. and West, eds.; Elsevier: Amsterdam, 1983, vol. 20, pp. 83 and references cited therein]. Mitoxantrone is a broad spectrum oncolytic agent, whose activity is similar to that of the anthracycline antibiotic doxorubicin.

Clinical trials have demonstrated that mitoxantrone has particularly promising activity in the treatment of advanced breast cancer, acute leukemia and lymphoma [Legha, Drugs of Today, 20, 629 (1984)]. Although animal studies have demonstrated a diminished cardiotoxicity in comparison to doxorubicin, some clinical cardiotoxicity has been observed also with mitoxantrone, mostly in patients previously treated with doxorubicin Stuart Harris et al., Lancet, 219, (1984) and references cited therein].

Ametantrone has been reported to be, in animals, about 10-fold less potent and cardiotoxic than mitoxantrone. Because a delayed toxicity is observed only with mitoxantrone after administration of the two drugs by the i.p. route to non-tumor bearing rats at equieffective antitumor dosages, it is suggested that the presence of the 5,8-dihydroxy substitution in mitoxantrone might be implicated in WO 97/02267 PCT/US96/10966 2 the delayed deaths [Corbett et al., Cancer Chemother. Pharmacol., 6, 161 (1981)].

In addition, both mitoxantrone and ametantrone have a remarkable myelodepressive toxicity and both compounds show cross-resistance to cell histotypes developing resistance against doxorubicin mediated by overexpression of glycoprotein P. Such a resistance, which is named multidrug resistance (MDR), involves a number of antitumor antibiotics, among which amsacrine and podophyllotoxinic In addition, both mitoxantrone and ametantrone have a remarkable myelodepressive toxicity and both derivatives, and it is one of the main reasons for therapeutic failures in the treatment of solid tumors with said antibiotics.

In an attempt to overcome the above mentioned drawbacks some chromophore modified anthracenediones have been prepared. For example, E. P. Patent Application 103.381 discloses 2 -aminoalkyl-5-aminoalkylamino substituted anthra[1,9-cd]pyrazol-6(2H)-ones (anthrapyrazoles) which are claimed to have antitumor activity. The antitumor activity of said compounds in a number of preclinical models has been reported by H.D. Hollis Showalter et al. Med.

Chem., 30, 121-131 (1987)]. However anthrapyrazoles are not devoid of toxic side effects, with severe leukopenia grade 3 and 4) and neutropenia grade 4) being dose limiting in phase I and phase II clinical trials with the anthrapyrazole CI-941 Smith et al., J. Clin. Oncol., 9, 2141-2147 (1991)]. Moreover a marked nephrotoxicity is associated with CI-941 treatment in the rat Campling and M.E.C. Robbins, Nephrotoxicitv, Peter H. Dekker Bach editor, pp. 345-352 (1991), New York; see Chemical Abstract 116: 294n (1992)] and these authors suggest that renal injury may be a clinical problem with anthrapyrazole therapy. In addition recent reports [Drugs of the Future, 17, 725 (1992); Judson, I.R. et al., Proc. Amer. Assoc. Cancer Res., 32, abstr. 1059 (1991)] indicate that the anthrapyrazole CI-941 induces irreversible cardiotoxicity in humans, although no symptoms of cardiac failure or acute cardiac events have been reported.

Furthermore, W094/06795 (31.03.94) describes aza-anthrapyrazole derivatives which are endowed with antitumor activity.

3 In the attempt to reduce the radical formation "in vivo" by eliminating the "quinonoid" structure, benzothiopyranoindazoles have been prepared

[H.D.

Hollis Showalter et al., J. Med. Chem., 31, 1527-1539 (1988)]. In these compounds a carbonyl group at C-6 position has been replaced by a sulphur atom. A compound of this series, CI-958, has been chosen for development toward clinical trials.

However, the search for newer active analogues is still highly desirable.

We have now discovered that the introduction of one nitrogen atom in the positions 7, 8, 9 or 10 of the above mentioned benzothiopyranoindazoles to provides 2H-pyrido[3',2':5, 6 ]thiopyrano[4, 3 ,2-cd]indazoles, 2Hpyrido[4',3':5,6]thiopyrano[4,3, 2 -cd]indazoles, 2Hpyrido[3'4':5,6]thiopyrano[4,3 ,2-cd]indazoles and 2Hpyrido[ 2 3 ':5,6]thiopyrano[4,3,2-cd]indazoles substituted in the positions 2 and 5, which have showed a marked antitumor activity.

15 BRIEF SUMMARY OF THE INVENTION The compounds of the invention have the general formula

D

X

wherein: v x S I T hydroxy (OH) or NR2(I)R3groups;

T

B

wherein: one of X, Y, Z or T is nitrogen and the others are a CH group; D is selected from the group consisting of nitro or -NH-A, wherein A is on its turn selected in the group consisting of hydrogen,

-CO-CH

2

-NR

2

R

3 C-Clo alkyl; C2- Ct alky having one or two substituents selected from the group consisting of OR, and -NR 2

R

3 C2-Ci1 alkyl interrupted by one or two oxygen atoms or by one

-NR

4 group, and said C2-Ci0 alkyl is optionally substituted by one or two hydroxy (OH) or -NR 2

R

3 groups; B is selected in the group consisting of C-Cio alkyl; C,-C,10 alkyl having one or R two substituents selected from the group consisting of OR, and -NR 2

R

3

C

2

-C

10 j alkyl interrupted by one or two oxygen atoms or by one -NR 4 group, and said C2- Clo alkyl is optionally substituted by one or two hydroxy (OH) or -NR 2

R

3 groups;

R

1 is selected from the group consisting of hydrogen, C1-C6 alkyl, -S(0 2

)R

5 C2-C6 alkyl optionally substituted by -NR 2

R

3

R

2 and R 3 may be the same or different and are selected from the group consisting of hydrogen, Ci-Clo alkyl, C2C 10 alkyl substituted with one or two hydroxy (OH) groups, or R 2 and R 3 taken together with the nitrogen atom to which they are linked form a 5- or 6-member aromatic or non-aromatic heterocyclic ring which optionally contains another heteroatom such as sulfur, oxygen or nitrogen;

R

4 is selected from the group consisting of hydrogen, C1-Clo alkyl, C2-C10 hydroxyalkyl, C2-C10 alkyl substituted with -NR 2

R

3

R

5 is selected from the group consisting of Ci-Clo alkyl, phenyl, phenylalkyl, as free bases and their salts with pharmaceutically acceptable acids.

The present invention also concerns the tautomeric forms, the single enantiomers 15 and disastereoisomers of the compounds of formula as well as mixtures thereof.

The present invention also concerns the non-toxic salts of the compounds of formula with acids acceptable for pharmaceutical and veterinary use such as those obtained by addition of inorganic acids like hydrochloric, hydrobromic, 20 sufuric, phosphoric, pyrophosphoric acid and/or of organic acids such as acetic, propionic, citric, benzoic, lactic, maleic, fumaric, succinc, tartaric, glutamic, aspartic, gluconic, ascorbic acids and the like.

The present invention further concerns a method of making the compounds of formula comprising: a) reacting a compound of formula (III):

NO

2 Y

XS

Zr (Il) O U 1 October 1999 wherein X, Y, Z and T are as defined above and U is selected from the group consisting of F or Cl, with a hydrazine of formula (IV):

H

2 N-NH-B'

(IV)

wherein B' has the same meanings as B is defined in formula or B' is a group that can be converted into B by removal of protective groups for the primary or secondary amines and hydroxy groups optionally present in to give a compound of formula (II): NO0 Y X

S

I(II)

z^ L (ID

B'

1 in which X, Y, Z, T and B' are as above defined.

The present invention still further concerns a pharmaceutical composition comprising the compound of formula and a pharmaceutically acceptable excipient.

The present invention also relates to a method of treating tumors in a patient in need of such treatment comprising administering an anti-tumor effective amount of the compound of formula The present invention further relates to use of the compound of formula as an anti-tumor compound.

DETAILED DESCRIPTION OF THE INVENTION In compounds the term "phenyl" means phenyl rings which can optionally contain substituents such as (C 1

-C

4 alkyl groups, CF 3 halogen atoms, nitro, amino, acetylamino, formylamino, dimethylamino, diethylamino, hydroxy, methoxy and ethoxy groups.

Preferred examples of C 1 -Co alkyl groups are methyl, ethyl, n-propyl, sec-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl.

Preferred examples of phenylalkyl is 4-methylphenyl When in compounds of '2mula A and B are a C 2

-C

10 alkyl interrupted by one or two oxygen atoms or 315.RS1 1 October 1999 WO 97/02267 WO 9702267PCT/US96/10966 by one -NR 4 group and optionally substituted by one or two hydroxy or -NR 2

R

3 groups, at least two carbon atoms are preferably interposed between said oxygen atoms and/or the -NR 4 and -NR 2

R

3 groups.

When in compounds of formula the -NR 2

R

3 substituent is a 5-6 member aromatic or not aromatic heterocyclic ring which may contain another heteroatom such as sulfur, oxygen and nitrogen, preferred examples of said heterocyclic rings are 1 -imidazolyl, 4-hydroxy-1 -imidazolyl, 2-imino-1 (3H)imidazolyl, 1 -pyrrolyl, 1 -tetrahydropyrrolyl, 1 -pyrazolyl, 4-morpholinyl, 1Ipiperidinyl, I -piperazinyl, I -(4-methyl )piperazinyl, I 4 -benzyl)piperazinyl.

The compounds of formula are those depicted in table 1, wherein D and B are as above defined.

TABLE 1

D

I

I

N-N

X Y Z T structure name of the heterocyclic system N S CHI CH CH N N-N. 2H-pyridol2',3':5,6]thiopyrano[43,2

B

_______cdJindazole N S CH CH N CHI N-N. 2H-pyrido[3',4':5,6jthiopyrano[43,2

B

_______cdjindazole

D

N

S

CH N CH CH N-N. 2H-pvndo[4',3':5,6thiopyrano[4,3,2.

B

WO 97/02267 PCT/US96/10966

D

N, S N CH CH CH N-NB 2H-pyrido[3',2':5,6]tiopyrano[4,3,2-

B

cd~indazole Preferred compounds are those according to formula wherein X, Y, Z and T are as above defined, D is a -NH-A group and A and B are independently selected from the group consisting of: residue of formula -(CH 2 )p-NH 2 wherein p is the integer 2 or 3; residue of formula -(CH 2 )p-NR 2

R

3 wherein p is as above defined and both R 2 and R 3 are methyl or ethyl or 2-hydroxyethyl; residue of formula -(CH 2 )p-NR 2

R

3 wherein p is above defined and R 2 is hydrogen and R 3 is methyl; residue of formula -(CH 2 )p-NR 2

R

3 wherein p is above defined and -NR 2

R

3 is 4-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-imidazolyl; residue of formula -(CH 2 )p-OH wherein p is as above defined; residue of formula -(CH 2 )p-NH-(CH 2 )q-OH wherein p and q are independently an integer selected from the group consisting of 2 or 3.

Other preferred compounds are those in which D is selected in the group consisting of nitro, amino and -NHCO-CH 2

-NH

2 and B is as just above defined.

Particularly preferred compounds of formula are those in which A and B are as previously defined and X or Z are nitrogen.

Even more particularly preferred compounds are those in which X or Z are nitrogen, D is a -NH-A group and A and B are independently a residue of formula -(CH 2 )p-NR 2

R

3 wherein p is as above defined and both R 2 and R 3 are methyl or ethyl or 2-hydroxyethyl.

The compounds of formula can be prepared by the reaction of a compound of formula (III): WO 97/02267 PCT/US96/10966 o U wherein X, Y, Z and T are as above defined and U is selected from the group consisting of F or Cl, with a hydrazine of formula (IV):

H

2 N-NH-B'

(IV)

wherein B' has the same meanings as B is defined in formula or B' is a group that can be converted into B by removal of protective groups for the primary or secondary amines and hydroxy groups optionally present in to give compound of formula (II): NO0 YZT Y

N-N

B'

in which X, Y, Z, T and B' are as above defined.

The removal of protecting groups optionally present gives the compounds of formula in which D is a nitro group.

Alternatively, said intermediate (II) is subjected to reduction of the nitro group to give the intermediate of formula (Ila):

NH

2 -x S z I I (Ila)

N-N

B'

which is then converted into the compounds of formula in which D is an amino group by removal of the protecting groups optionally present.

WO 97/02267 PCT/US96/10966 8 Alternatively, the intermediate (lla) is converted into the compounds of formula in which D is a -NH-A group and A has the meanings above defined, with the exception of hydrogen and -CO-CH 2

-NR

2

R

3 by condensation of the amino group with a reactant of formula L-A'

(V)

in which A' has the same meanings as A is defined in formula or A' is a group that can be converted into A by removal of protective groups for the primary or 0o secondary amines or hydroxy groups optionally present in A' and L is an atom selected in the group of chlorine, bromine and iodine or is a -O-Tosyl group, and subsequent removal of protective groups optionally present on the side chains A' and B'.

Alternatively, A' is a group of formula in which on its turn A" is selected in the group consisting of CI-C 9 alkyl; Ci-C 9 alkyl having one or two substituents selected from the group consisting of OR, and -NR 2

R

3

C

1

-C

9 alkyl interrupted by one or two oxygen atoms or by one -NR 4 group, and said C 1

-C

9 alkyl is optionally substituted by one or two hydroxy (OH) or -NR 2

R

3 groups, and in which the hydroxy or the amine group can optionally be protected with conventional protective groups. In this case, L is a OH group, a halogen atom (chlorine, bromine or iodine) or a suitable leaving group well known to the skilled artisan. Such a reaction, followed by the removal of the protecting groups optionally present, gives the compounds of formula in which A is a -CO-CH 2

NR

2

R

3 group. They on their turn give the other compounds of formula by reduction of the amidic moiety to amine.

Protective groups for the primary and/or secondary amines optionally present in A' and/or B' which can advantageously be used for the preparation of compounds of formula are represented by (Ci-C 3 )acyl derivatives (preferably acetyl derivatives),

(C

1 -C4)alkoxycarbonyl derivatives (preferably tertbutoxycarbonyl derivatives) and by (C7-Clo)aralkyloxycarbonyl derivatives (preferably benzyloxycarbonyl derivatives).

The reaction of compounds (III) with the hydrazines (IV) can be performed by reacting compounds (III) with a stoichiometric amount of hydrazines (IV) or an WO 97/02267 PCT/US96/10966 9 excess of hydrazines The reaction is usually performed in an inert solvent such as methylene chloride, chloroform, 1,1,1-trichloroethane, dimethoxyethane, tetrahydrofuran, dimethylsulfoxide, dimethylformamide, pyridine, picoline and mixtures thereof, or if it is desired, using compound (IV) itself as the solvent, optionally in the presence of an inorganic base such as an alkaline or alkalineearth carbonate or hydrogen carbonate or an organic base such as a trialkylamine, at a temperature from -20°C to the reflux temperature of the solvent. Preferably, the reaction is carried out in a solvent such as dimethylformamide, pyridine, tetrahydrofuran, dimethylsulfoxide or tetramethylethylenediamine, using 1 to 10 equivalents of compound (IV) for 1 equivalent of compound (III) and working at a temperature ranging from 5°C to 0

C.

If necessary, the removal of the protective group for the primary and/or secondary amino functions is carried out following the procedures well known to those skilled in the art. Useful teachings can be found in Green, Wuts, "Protective Groups in Organic Synthesis", second Edition, John Wiley Sons, 1991.

For example, the removal of the N-(tert-butoxycarbonyl) protective group can be performed by treatment of the compound with an excess of anhydrous or aqueous hydrochloric acid in a solvent such as a (Ci-C4)alkanol, dichloromethane, chloroform or mixtures thereof, at a temperature of 0=C to the reflux temperature of the solvent and for a time ranging from several minutes to a few hours. Preferably the reaction is performed in ethanol or in chloroform using from 10 to 20 molar equivalents of anhydrous hydrochloric acid at a temperature of from 20°C to 50°C, and is generally complete in four hours.

The reaction of reduction of the nitro group of compounds (II) to give compounds (Ila) is performed by catalytic hydrogenation, treating in hydrogen atmosphere a compound in a suitable solvent such as glacial acetic acid, an alkanol such as methanol or ethanol, an ether such as 1,4-dioxane, tetrahydrofuran or a solvent such as methylene chloride or chloroform, in the presence of catalytic amounts (from 0.1 to 0.3 equivalents) of a catalyst such as palladium on carbon, Nickel Raney or 10% platinum on carbon and at a pressure of from atmospheric pressure to 100 psi.

WO 97/02267 PCT/US96/10966 Preferably, the reaction is carried out in glacial acetic acid, using 10% palladium on carbon as a catalyst and at a pressure of about 70 psi.

Alternatively, the reduction of the nitro group of compounds (11) to give compounds (Ila) may be performed following other methods known in the art, such as the use of FeSO 4 in basic conditions (ammonia or sodium hydroxide) or the use of metals or salts thereof, optionally in acidic conditions. Suitable examples of such reducing agents are SnCI 2 SnCI 2 in the presence of hydrochloric acid or Zn in the presence of hydrochloric acid.

The alkylation of intermediates (Ila) with intermediates of formula L-A' to give 0o the compounds of formula is performed by reacting the intermediate (Ila) with a molar excess of the reactant of formula in an inert solvent such as benzene, toluene, chlorobenzene and the like, or an alcohol such as isopropanol or in a nitrile such as acetonitrile, and in the presence of an inorganic base such as an alkaline or alkaline-earth carbonate or hydrogencarbonate or an organic base such as trialkylamine. Preferably, the reaction is carried out in an aromatic solvent such as toluene, in the presence of potassium carbonate and at a temperature ranging from room temperature to the reflux temperature of the solvent.

The acylation reaction of intermediates (Ila) with compounds of formula L-CO-A" is usually performed, when L is a OH group, in the presence of a condensing agent such as dicyclohexylcarbodiimide and the like, at a temperature ranging from -10°C to room temperature and in an inert solvent such as an ether (tetrahydrofuran, diethyl ether and the like).

The reduction of amidic moiety into amine can be preferentially performed in an inert solvent such as toluene or benzene, at a temperature ranging from room temperature to the reflux temperature of the solvent and in the presence of a reducing agent such as Red-AI, LiAIH 4 or other suitable hydrides.

The intermediates (III) can be prepared by cyclization of the compounds of formula (VI): WO 97/02267 PCT/US96/10966 11

U

.T CO 2

H

IX I I (VI) "X S

NO

2 in which X, Y, Z, T and U are as above defined.

Said reaction can be performed using different methods known in the art, such as for example: i) transforming the carboxylic acid moiety into an acyl chloride by reaction with thionyl chloride, for example, and subsequently performing a Friedel-Craft reaction in the presence of a Lewis acid such as alluminum trichloride, in a suitable solvent such nitrobenzene and at a temperature ranging from -10°C to ii) cyclizing the compounds (VI) in the presence of fuming sulfuric acid sulfur trioxide), at a temperature ranging from room temperature to 150°C; iii) cyclizing the compounds (VI) in the presence of polyphosphoric acid silyl ester (PPSE) and of phosphorus pentoxide, at a temperature ranging from to 21 0 0C.

The compounds of formula (VI) can be obtained starting from the compounds of formula (VIII): ZT CO 2

H

X Q X Q

(VIII)

in which X, Y, Z and T are as above defined and Q is a chlorine atom or a diazo group

-N

2 CI", by reaction with a compound of formula (VII): WO 97/02267 PCT/US96/10966 12

U

H- (VII)

NO

2 in which U is as above defined.

When X or Z are nitrogen, then Q is preferentially chlorine; when Y or T are nitrogen, Q is preferentially a -N 2 *Cl group.

When Q is chlorine, the reaction is preferentially performed in a suitable solvent such as a ketone (acetone, methyl ethyl ketone and the like) and at a temperature ranging from room temperature to the boiling point of the solvent.

When Q is a -N2*ClI group, the compounds (VIII) are prepared "in situ", following 0o the methods known in the art, by reaction of the amine derivatives

(IX):

zT

CO

2

H

y NH

(IX)

X

NH

2 with sodium nitrite in the presence of hydrochloric acid at a temperature ranging from -10°C to 10°C. The compounds (VIII) so prepared are then reacted without isolation with the compounds (VII) at a temperature ranging from 0°C to obtaining the compounds of formula (VI).

Alternatively, compounds (VIII) in which Q is a -N 2 'CI can be reacted without isolation with equimolar amounts of O-ethylxanthic acid, potassium salt. The resulting intermediates give compounds (VI) by treatment with sodium ethoxide in ethanol and subsequently with 2 4 -dichloronitrobenzene.

The compounds of formula (VIII) when Q is a chlorine atom or the compounds of formula (IX) are commercial product or can be prepared starting from commercial product following methods known in the art, such as for example those described in: Ross, C, 1816-21 (1966): synthesis of 4-chloro nicotinic acid; Winn, M. et al., J. Med. Chem., 36, 2676-88 (1993); WO 97/02267 PCT/US96/10966 13 Fibel, C.R. et al., 70, 3908 (1948): synthesis of 3-amino isonicotinic acid.

BIOLOGICAL EVALUATION OF THE COMPOUNDS OF THE INVENTION The evaluation of the biological activity for the compounds of this invention was performed "in vitro" and "in vivo" following the protocols developed by the U.S.

National Cancer Institute.

The evaluation of the "in vitro" cytotoxic activity of the compounds of the invention was performed using the following cell lines: a murine sarcoma (S- 180) and its subline expressing multidrug resistance (S-180/A-10), a leukemia to (L1210), a human colon adenocarcinoma cell line (LoVo) isolated from a metastatic nodule and its subline expressing multidrug resistance. This latter subline is resistant to a number of antitumor agents, among which are doxorubicin, VP-16 and vincristine. This subline (named LoVo/DX) shows reduced accumulation of doxorubicin and overexpression of a protein (Grandi, Geroni, Giuliani, British J. Cancer, (1986), 54, 515). The compounds were tested according to the MTT assay (Mosman, T. "Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assay", J. Immunol. Methods, (1983), 65, 55-63; Green, L.M., "Rapid Colorimetric Assay for Cell Viability; Application to the Quantitation of Cytotoxic and Growth Inhibitory Lymphokines", J. Immunol. Methods, (1984), 257-268) in comparison with mitoxantrone and doxorubicin.

The pharmacological data for some representative compounds of the present invention are reported in Tables II and III. A comparison with the prior art compound C1941 and with doxorubicin and mitoxantrone is also provided in Table II. The columns headed L1210, S180 LoVo and LoVo/Dx contain the values against said tumor cell lines, as above described.

In general, representative compounds of this invention exhibited a high cytotoxicity in all the cell lines tested. When mitoxantrone was tested in the LoVo/DX cell line, a resistance index RI (defined as the ratio of the ICso for the resistance cell line to the ICsofor the sensitive cell line) as high as 22.5 was found, showing that this subline does have an acquired resistance to mitoxantrone. On the other hand, representative compounds of this invention, when tested in the same resistant subline, show no cross resistance with WO 97/02267 PCT/US96/10966 14 mitoxantrone. The "in vitro" evaluation of representative compounds of this invention suggests that they are able to retain a high activity also in the resistant cell line, while the prior art compound CI-941 loses completely its activity.

Studies of the biological activity "in vivo" of representative compounds of the invention were performed using the P388 murine leukemia model. P388 murine leukemia cells were intravenously (iv) injected in CD2F1 mice. Treatment was initiated approximately 24 hours after tumor transplantation and dosages of the drug were administered iv (P388 iv/iv) according to preestablished protocols, usually at 3-day intervals. The studies were done over a 60-day period and the 1o date of death for each animal was recorded. The TIC was determined using the mean survival time (MST) for each group according to the formula TIC [(MST treated) (MST control)] x 100 Representative compounds of this invention were able to increase the survival time of treated animals, leading to high TIC values at well tolerated dosages.

Since representative compounds of this invention show good results against this significant "in vivo" model of murine P388 leukemia, which is considered to be predictive of antitumor activity in humans, the compounds disclosed herein are expected to be operative against human leukemias and solid tumors sensitive to treatment with antitumor antibiotics.

The compounds of the invention may therefore be used as active ingredients of therapeutic compositions to induce regression and/or palliation of cancers in mammals when administered in amounts ranging from about 1 mg to about 0.4 g per kilogram of body weight. A preferred dosage regimen would be from about 1 mg to about 50 mg per kilogram of body weight per day. Unit dosage may be employed so that from about 70 mg to about 3.5 g of the active compound for a subject of about 70 kg of body weight are administered in a 24-hour period. The dosage may be adjusted to be compatible to other treatment regimens, such as radiation therapy.

The pharmaceutical composition may be in the form of tablets, capsules, gel capsules, suppositories, lyophilized powde and solutions for intravenous administration.

WO 97/02267 PCT/US96/10966 The invention is illustrated by the following non-limiting examples and variations which are readily apparent to those skilled in the art.

EXPERIMENTAL

PART

Preparation 1 2 2 -nitro-5-chloro)thiophenoxvnicotinic acid (compound

X

N)

A solution of 2-nitro-5-chlorobenzenethiol (0.26 g) in acetone (2 ml) was added to 2-chloronicotinic acid (0.11 The suspension was refluxed for 5 hrs and the resultant mixture was cooled to room temperature. The bright yellow precipitate was collected by filtration, washed with acetone to yield a yellow solid (0.12 g), 0o m.p. 248-249*C.

'H NMR in d6-DMSO 8.39 ppm (dd, J 1.7 Hz, 4.7 Hz, 1H); 8.27 ppm (dd, J 1.7 Hz, 7.9 Hz, 1H); 8.10 ppm J 8.7 Hz, 1H); 7,85 ppm J 2.3 Hz, 1H); 7.76 ppm (dd, J 2.3 Hz, 8.7 Hz, 1H); 7.30 ppm (dd, J 4.7 Hz, 7.7 Hz, 1H).

Anal. Calcd. for C 12

H

7

CIN

2 0 4 S: C 46.39, H 2.27, N 9.01. Found: C 46.39, H 2.27, N 8.80.

Preparation 2 6-chloro-9-nitro-5H-1 lbenzothiopvranof2.3:blpvridin-5-one (compound (I11), X N) Route 1 (with fuming sulfuric acid) 0.645 g of 2 2 -nitro-5-chloro)thiophenoxynicotinic acid were added to 2 ml of fuming sulfuric acid (18-24% sulfur trioxide) and the mixture was placed in an oil bath preheated to 75"C. The solution was heated at 125-130 0 C for 1.25 hrs. The mixture was removed from the oil bath, cooled to room temperature and poured over ice water (150 ml). The yellow precipitate was collected by filtration, washed with water and dried, to give 0.60 g of a solid. This material was dissolved in dimethyl formamide (DMF, 11 ml) which on cooling immediately led to a yellow crystalline fluffy solid. This solid was collected by filtration and washed with diethyl ether to remove residual DMF, yielding 0.54 g of product, m.p. 267-270 0

C.

1H NMR in CDCI 3 8.84 ppm (dd, J 1.77 Hz, J 4.60 Hz, 1H); 8.60 ppm (dd, J 1.77 Hz, J 8.05 Hz, 1H); 8.51 ppm J 8.80 Hz, 1H); 7.68 ppm J 8.80 Hz, 1H); 7.51 ppm (dd, J 4.6 Hz, J 8.00 Hz, 1H).

Route 2 (with thionyl chloride/ alluminum trichloride) WO 97/02267 PCT/US96/10966 16 A mixture of 2 2 -nitro-5-chloro)thiophenoxynicotinic acid (5 toluene (27 ml) and thionyl chloride (6 ml) was heated at reflux for 1.5 hrs. Upon cooling, the acid chloride separated as yellow needles. The resultant mixture was concentrated to dryness by distillation and a yellow crystalline solid remained.

Nitrobenzene (25 ml) was added and the suspension was cooled in an ice bath for 0.5 hrs. Alluminum chloride (2 g) was slowly added while keeping the temperature below 35C. The mixture darkened and it was stirred at room temperature for 20 hrs. The dark black suspension was poured into ice water (130 ml) and the mixture was stirred for 1 hr. The aqueous layer was removed to by decantation, methanol (100 ml) was added to the nitrobenzene and the resultant solid was collected by filtration. Addition of methanol (200 ml) to the filtrate led to additional product, total weight 3.1 g. For purification, the crude material was recrystallized from ethylene glycol monoethyl ether to yield a yellow fluffy solid, m.p. 265-270"C, identical in its 'H-NMR spectrum to the product obtained following route 1.

Route 3 (with polyphosphoric acid silvl ester. PPSE) A mixture of PPSE (2 g) and phosphorus pentoxide (0.25 g) was heated in an oil bath to 210°C. 2 2 -nitro-5-chloro)thiophenoxynicotinic acid (0.10 g) was added to the hot mixture and the mixture was held at this temperature for 20 minutes.

The hot mixture was quenched into cold hydrochloric acid 6 N (6 ml) and the resultant mixture was allowed to stand overnight. After neutralization with sodium hydroxide, the solid was collected by filtration and dried to give 0.085 g of a solid. The crude material was heated in ethyl acetate and filtered while hot to remove some brownish insoluble material. Removal of the solvent led to product (0.045 g) identical to that prepared following routes 1 and 2.

Preparation 3 3 2 -nitro-5-chlorothiophenoxv)isonicotinic acid (compound

(VI).

Y N) A solution of 3-amino-4-carboxypyridine (1.4 sodium nitrite (0.81 g) and aqueous sodium hydroxide 2.9 M (13.3 ml) was added to a solution of aqueous hydrochloric acid 4.3 M (9.8 ml) while the temperature was maintained at 0-5 0

C.

The mixture was stirred for 10 minutes and then added dropwise over the course of 2 hrs to a stirred mixture of 5-chloro-2-nitrothiophenol (2.34 g) and i;l- 'iiiiii:---i- WO 97/02267 PCT/US96/10966 17 sodium hydroxide (2.63 g) in water (21 ml) maintained at about 53°C (a brisk nitrogen evolution occurred). The mixture was stirred for 15 minutes, cooled to room temperature and filtered. The fltrate was made strongly acidic (pH 2) with hydrochloric acid 12 M and the precipitated solid was collected by filtration and dried to yield a brownish-orange solid (3 The solid was washed with dichloromethane:acetonitrile (30 ml) and dried to leave crude product (1.2 g).

Crystallization from ethyl cellosolve led to the pure product, m.p. 280-281

C.

1H NMR in d6-DMSO 8.77 ppm J 4.8 Hz, 1H); 8.67 ppm 1H), 8.23 ppm J 8.8 Hz, 1H); 7.77 ppm J 4.8 Hz, 1H); 7.56 ppm (dd, J 8.8 Hz, J 2.2 Hz, 1H); 6.99 ppm J 2.1 Hz, 1H).

Preparation 4 6-chloro-9-nitro-5H-[1 lbenzothiopyrano2. (compound (III), Y N) A mixture of the compound of the preparation 3 (0.45 g) and thionyl chloride ml) was heated at reflux for 1.5 hrs. The mixture changed to a dark reddish coloration. The mixture was concentrated to dryness by distillation to yield a dark red amber solid. Nitrobenzene (3.5 ml) was added and the suspension was cooled in an ice bath for 0.5 hrs. Alluminum chloride (1.02 g) was added slowly and the dark mixture was allowed to warm to room temperature and then stirred at 80-90°C for 15 hrs. The dark suspension was poured over crushed ice (25 ml) and the mixture was stirred for 1 hr. The aqueous layer was decanted and methanol (25 ml) was added to the nitrobenzene layer. The resultant solid was collected by filtration to yield crude product (300 mg) as a dark grey solid. The solid was boiled with chloroform (75 ml) and filtered to remove some insoluble material. Concentration of the filtrate led to the pure product (75 mg).

H NMR in CDCI 3 9.02 ppm 1H); 8.79 ppm J 5.2 Hz, 1H); 8.54 ppm (d, J 8.8 Hz, 1H); 8.09 ppm J 5.2 Hz, 1H); 7.70 ppm J 8.8 Hz, 1H).

Preparation 5 4 2 -nitro-5-chloro)thiophenoxy nicotinic acid (compound

Z

A solution of 2-nitro-5-chlorothiophenol (1.09 g) in acetone (12 ml) was added to 4-chloronicotinic acid (0.85 The yellow coloration of the thiol quickly disappeared and the mixture was refluxed f. r 1 hr. Upon cooling to room temperature, the product was filtered and washed with acetone to yield the title compound as pale yellow solid (1.72 m.p. 228-229 0

C.

WO 97/02267 PCT/US96/10966 18 1H NMR in d6-DMSO 9.05 ppm 1H); 8.50 ppm J 5.9 Hz, 1H); 8.23 ppm J 8.7 Hz, 1 7.98 ppm J 2 Hz, 1 7.92 ppm (dd, J 8.7 Hz, J 2.2 Hz, 1H); 6.98 ppm J 5.9 Hz, 1H).

Preparation 6 6 -nitro-9-chloro-10H-1i benzothiopyrano2 3-clpvridin-1 0-one (compound (III). Z N) Thiophenoxynicotinic acid (0.50 g, preparation 5) was added to fuming sulfuric acid (18-24% sulfur trioxide, 3 ml) and the mixture was placed in an oil bath which was preheated to 40"C. The dark reddish amber solution was heated to °C during 10 minutes and kept at this temperature for 20 minutes. The cooled 0o mixture was poured over ice-water (25 ml) and neutralized with solid sodium bicarbonate. The resultant bright yellow solid was collected by filtration and dried, obtaining 0.38 g of crude product. The solid was boiled in chloroform ml) and filtered to remove some starting material. Concentration of the filtrate led to the pure product (0.33 m.p. 220-222 0

C.

'H NMR in CDCIl 9.43 ppm 1H); 8.74 ppm J 5.5 Hz, 1H); 8.50 ppm (d, J 8.8 Hz, 1H); 7.70 ppm J 8.8 Hz, 1H); 7.51 ppm J 5.5 Hz, 1H).

Preparation 7 N'-12-(2-dimethvlamino)ethyll-5-(2'-(N-tertbutoxvcarbonvl)amino acetylamino)-2H-pvridor3',2':5,61thiopvrano4, 3,2-cdlindazole Dicyclohexylcarbodiimide (210 mg) was slowly added to a magnetically stirred solution of _N,N-dimethyl-5-amino-2H-pyrido[3',2':5,6]thiopyrano[4,3,2cd]indazole-2-ethanamine(300 mg) of example 2 and N-BOC-glycine (168 mg) in dry tetrahydrofuran (8 ml) at 0°C. The mixture was stirred at 0-5 °C for 20 hr and the cyclohexylurea which precipitated was removed by filtration. The filtrate was refrigerated overnight and additional urea was removed by filtration. The filtrate was concentrated to dryness by rotary evaporation and the brownish red solid was collected (434 mg). This material was recrystallized from acetonitrile/chloroform 4:1 to give the product as a yellow solid (240 mg).

Purification was best accomplished by column chromatography over silica gel using gradient elution by ethyl acetate methanol as eluent, commencing with 5:1 and gradually changing to 1:1. Removal of the eluents led to the purified product (120 mg, m.p. 173-175 0

C).

WO 97/02267 PCT/US96/10966 19 'H-NMR in CDCIs 8.34 ppm (dd, J 1.7 Hz, 4.7 Hz, 1H); 8.23 ppm (dd, J 1.7 Hz, 7.8 Hz, 1H); 7.60 ppm (broad s, 1H); 7.49 ppm J 8.7 Hz, 1H); 7.15 ppm J 4.7, 7.8 Hz, 1H); 6.97 ppm J 8.8 Hz, 1H); 5.21 ppm (broad d, 1H); 4.39 ppm J 6.9 Hz, 2H); 4.00 ppm J 6.0 Hz, 2H); 2.82 ppm J 8.9 Hz, 2H); 2.33 ppm 6H); 1.49 ppm 9H).

Preparation 8 N'-[2-(2-dimethylamino)ethyll-5-(2'-(N-tertbutoxycarbony)amino acetylamino)-2H-pyridor3',4':5,61thiopvranof 4 3,2-cdlindazol According to the procedure described in preparation 7, the title compound was prepared.

1 H-NMR in CDCI 3 9.17 ppm 1H); 8.31 ppm J 5.4 Hz, 1H); 7.70 ppm (broad s, 1H); 7.40 ppm J 8.8 Hz, 1H); 7.73 ppm J 5.4 Hz, 1H); 7.00 ppm J 8.8 Hz, 1H); 5.21 ppm (broad s, 1H); 4.40 ppm J 6.9 Hz, 2H); 3.99 ppm J 6.0 Hz, 2H); 2.84 ppm J 6.9 Hz, 2H); 2.33 ppm 6H); 1.48 ppm 9H).

Example 1 N,N-dimethvl-5-nitro-2H-pyridor3'.2':5.61thiopyrano[4 3.2cdlindazole-2-ethanamine (X N) A suspension of 6-chloro-9-nitro-5H-[1]benzothiopyrano[2,3:b]pyridine-5-one g) in DMF (25 ml) under a nitrogen blanket was cooled in an ice bath and 2o N-(2-dimethylaminoethyl) hydrazine (1 g) was added dropwise. The coloration changed from yellow to bright orange. The suspension was stirred for 15 hrs at room temperature and the mixture was quenched over ice-water. The pH was adjusted to 10.5-11 by addition of a saturated solution of potassium carbonate.

The resultant mixture was extracted with chloroform (2x100 ml) and the chloroform layer was washed with cold water (150 ml) and then with brine (2x150 ml). The chloroform was dried over magnesium sulfate, the drying agent removed by filtration and the filtrate concentrated to yield the product as a golden brown solid (2.5 This material crystallized readily from acetonitrile, m.p. 173-174*C.

H NMR in CDC3 8.55 ppm (dd, J 1.4 Hz, J 4.6 Hz, 1H); 8.41 ppm (dd, J 1.4 Hz, J 7.9 Hz, 1H); 8.23 ppm J 9.2 Hz, 1H); 7.33 ppm (dd, J 4.6 Hz, J 7.8 Hz, 1H); 7.05 ppm J 9.2 Hz, 1H); 4.48 ppm J 6.6 Hz, 2H); 2.88 ppm t, J 6.6 Hz, 2H); 2.30 ppm 6H).

WO 97/02267 PCT/US96/10966 Example 2 N.N-dimethvl-5-amino-2H-pyridor3'.2':5.61thiopvrano[4,3.2cdlindazole-2-ethanamine (X N) A mixture of the nitro analogue of example 1 (2.12 g) and 10% palladium/C (0.35 g) in glacial acetic acid (38 ml) was placed in a Parr bomb and hydrogenated for 18 hrs at about 100 psi. The mixture was filtered through celite and concentrated to yield a dark amber oil which was dissolved in chloroform (200 ml). This solution was washed with 5% aqueous ammonium hydroxide (200 ml), water (200 ml) and brine (2x200 ml). The chloroform layer was dried over sodium sulfate, the drying agent removed by filtration and the filtrate 0o concentrated to yield the product as a bright reddish-yellow solid (1.7 This was crystallized from acetonitrile to yield pure product (1.1 m.p. 184-186 0

C.

1H NMR in CDCI 3 8.28 ppm (dd, J 1.6 Hz, J 4.7 Hz, 1H); 8.16 ppm (dd, J 1,6 Hz, J 7.8 Hz, 1H); 7.10 ppm (dd, J 4.7 Hz, J 7.8 Hz, 1H); 6.86 ppm (d, J 8.6 Hz, 1H); 6.78 ppm J 8.6 Hz, 1H); 4.35 ppm J 7.1 Hz, 2H); 3.39 ppm 2H); 2.81 ppm 7.1 Hz, 2H); 2.24 ppm 6H).

Analogously, starting from the appropriate intermediate (example 3) the following compound was prepared (Y N): N,N-dimethyl-5-amino-2H-pyrido[4',3':5,6]thiopyrano[4,3,2-cd]indazole-2ethanamine, 'H NMR in CDC13 8.46 ppm 1H); 8.34 ppm J 5.2 Hz, 1H); 7.76 ppm (d, J 5.2 Hz, 1H); 6.88 ppm J 6.5 Hz, 1H); 6.80 ppm J 8.7 Hz, 1H); 4.37 ppm 2H); 3.38 ppm (br s, 2H); 2.83 ppm 2H); 2.31 ppm 6H).

Example 3 NN-dimethvl-5-nitro-2H-yvridof4',3':5,61thiopranor4.3.2cdlindazole-2-ethanamine (Y N) A suspension of the compound of preparation 4 (0.10 g) in DMF (0.8 ml) under a nitrogen blanket was cooled in an ice bath and N-(2dimethylaminoethyl)hydrazine (0.043 g) was added dropwise. The colour changed to bright orange and the suspension was allowed to stir at room temperature for 12 hrs. The mixture was quenched over ice water and basified with aqueous sodium bicarbonate. The resultant solid was collected by filtration and dried to yield the pure product (0.035 m.p. 205-206 0

C.

WO 97/02267 PCT/US96/10966 21 1 H NMR in CDC13 8.83 ppm 1H); 8.59 ppm J 5.2 Hz, 1H); 8.30 ppm (d, J 9.3 Hz, 1H); 8.10 ppm J 5.1 Hz, 1H); 7.11 ppm J 9.2 Hz, 1H); 4.52 ppm J 6.5 Hz, 2H); 2.90 ppm J 6.5 Hz, 2H); 2.31 ppm 6H).

Example 4 N.N-dimethyl-5-nitro-2H-pyridor3',4':5,61thiopyranof43,2cdlindazole-2-ethanamine (Z N) N-(2-dimethylaminoethyl)hydrazine (25 mg) was added to a suspension of the compound of preparation 6 (60 mg) in DMF (0.5 ml). The resultant orange suspension was allowed to stir for 15 hrs at room temperature. The mixture was added to cold water (5 ml) and treated with aqueous sodium bicarbonate. The to orange solid was collected by filtration, washed thoroughly with water and dried to yield 57 mg of the product. The sample was recrystallized from acetonitrile.

m.p. 214-215*C.

'H NMR in CDCI 3 9.40 ppm 1H); 8.52 ppm J 5.5 Hz, 1H); 8.27 ppm (d, J 9.3 Hz, 1H); 7.46 ppm J 5.6 Hz, 1H); 7.12 ppm J 9.3 Hz, 1H); 4.51 ppm J 6.5 Hz, 2H); 2.91 ppm J 6.5 Hz, 2H); 2.27 ppm 6H).

Example 5 NN-dimethvl-5-amino-2H-pyridor3',4':5, 6thioovranor4 3 2cdlindazole-2-ethanamine (Z N) A mixture of the compound of example 4 (0.25 g) and 10% palladium/C (0.042 g) in glacial acetic acid (4.5 ml) was placed in a Parr bomb and hydrogenated for 18 hrs at about 70 psi pressure. The mixture was concentrated to dryness, diluted with 5% aqueous ammonium hydroxide:chloroform and then filtered over celite. The chloroform layer was separated from the filtrate and washed with water and brine. The chloroform was removed by rotary evaporation to yield the product (0.15 g).

'H NMR in CDCI 3 9.10 ppm 1H); 8.25 ppm J 5.4 Hz, 1H); 7.12 ppm (d, J 5.4 Hz, 1H); 6.88 ppm J 8.7 Hz, 1H); 6.78 ppm J 8.7 Hz, 1H); 4.35 ppm J 7.0 Hz, 2H); 3.34 ppm (br s, 2H); 2.82 ppm J 7.0 Hz, 2H); 2.30 ppm 6H).

Example 6 N'-[2-[2-(dimethvlamino)ethvl-2H-pyridof3',2':5,61thiopyranof4.32cdlindazol-5-vll-N,N-dimethvl-l,2-ethanediamine (X N) A mixture of the amino analogue of preparation 4 (0.50 2- (dimethylamino)ethyl bromide hydrobromide (0.98 g) and potassium carbonate (1.37 g) in toluene (15 ml) was refluxed for 15 hrs. The mixture was cooled and WO 97/02267 PCT/US96/10966 22 the residue which was collected by filtration was triturated with hot acetonitrile.

The insoluble material was removed by filtration through a celite bed. The filtrate was concentrated to 15 ml and allowed to stand overnight. The product was collected by filtration as a golden brown material (0.36 This crude material was dissolved in hot acetonitrile (10 ml), cooled to room temperature and placed in the refrigerator overnight. The product as golden brown needles was collected by filtration (0.26 m.p. 110-111 C.

1H NMR in CDCI 3 8.27 ppm (dd, J 1.6 Hz, J 4.7 Hz, 1H); 8.16 ppm (dd, J 1.6 Hz, J 7.7 Hz, 1H); 7.09 ppm (dd, J 4.7 Hz, J 7.7 Hz, 1H); 6.92 ppm (d, J 8.7 Hz, 1H); 6.87 ppm J 7.1 Hz, 1H); 4.37 ppm J 7.0 Hz, 2H); 3.25 ppm J 5.9 Hz, 2H); 2.83 ppm J 7.0 Hz, 2H); 2.60 ppm J 5.9 Hz, 2H); 2.32 ppm 6H); 2.34 ppm 6H).

Example 7 N'-r2-r2-(dimetlnoethyll-2H-pvridor3' 2':561thiopyrano4,3,2cdlindazol-5-yl-1,2-ethanediamine (X N) A commercial Red-Al solution (3.4 M in toluene, 0.6 ml) was added dropwise over a period of 3 minutes to a stirred suspension of the compound of the preparation 12 (100 mg) in toluene (1 ml) held at 70 The resultant bright red solution was heated for an additional 5 hrs at 70-75 cooled to room temperature and treated cautiously with a saturated aqueous ammonium chloride solution. The yellow suspension was diluted with dichloromethane (3 ml) and the mixture was filtered through a celite bed. The organic layer was dried over magnesium sulfate and concentrated to yield the crude product (100 mg). This material was purified by column chromatography over silica gel eluting sequentially with 2, 4, 8, 10 and 20 methanol in dichloromethane. The center eluents were concentrated to yield the product as a yellow solid (50 mg, m.p.

77-80 'H-NMR in CDCI 3 8.29 ppm (dd, J 1.7, 4.7 Hz, 1H); 8.17 ppm (dd, J 1.7, 7.8 Hz, 1H); 7.11 ppm J 4.7, 7.8 Hz, 1H); 6.91 ppm J 8.8, 8.8 Hz, 2H); 4.86 ppm (broad s, 1H); 4.36 ppm J 7.0 Hz, 2H); 3.36 ppm (broad s, 4H); 2.82 ppm J 6.9 Hz, 2H); 2.32 ppm 6H); 1.48 ppm 9H).

Dry hydrogen chloride was briefly passed through a solution of the title compound (50 mg) in chloroform (0.5 ml). Upon removal of the chloroform, the hydrochloride salt of the product was obtained as an orange solid (43 mg).

WO 97/02267 WO 9702267PCTIUS96/1096 23 'H-NMR in D,0 8.23 ppm (dd, J 1.4, 4.8 Hz, 1 8.09 ppm (dd, J 1.4, 7.8 Hz, I 7.26 ppm (in, J 4.9, 7.8 Hz, 1 7.13 ppm J 8.8 Hz, 1 7.08 ppm J 8.8 Hz, 1 4.71 ppm J 5.9 Hz, 2H); 3.79 ppm J 5.9 Hz, 2H); 3.59 ppm J 6.2 Hz, 2H); 3.34 ppm J 6.2 Hz, 2H); 3.05 ppmn 6H).

Examrole 8 N'-[2-r2-(dimethylamino)ethvl-2H-rvrido3 .61thioov rano[4.3.2cdlindazol-5-vl-1 .2-ethanediamine (Z N) According to the procedure of example 7, starting from the intermediate of preparation 8, the title compound as hydrochloride salt is prepared.

'H-NMR in D 2 0 9.01 ppm (s,l1H); 8.39 ppm J =6.5 Hz,lIH); 7.90 ppm J 6.5 Hz, 1 7.39 ppm J 8.9 Hz, 1 7.27 ppm J 8.9 Hz, 1 4.86 ppm J 5.9 Hz, 2H); 3.88 ppm J 5.8 Hz, 2H); 3.62 ppm J 6.2 Hz, 2H); 3.36 ppm J 6.2 Hz, 2H); 3.09 ppm 6H).

Examrle 9 compounds (1) Following the methods described in example 6 or in example 7, starting from the appropriate amine analogues obtained according to example 5 and from the appropriate aminoalkyl halide (method of example 6) or alternatively from aminoacids or derivative thereof (method of example the following compounds of formula are prepared: 2 -[2-(diethylamino)ethyl]-2H..pyrido[3,2:.5,6]thiopyrano[4, 3,2-cdjindazol- 5-yl]-N, N-dimethyl-1 ,2-ethanediamine; 2 2 -(methylamino)ethyI]-2H-pyrido[3',2':5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-N,N-dimethy-1 ,2-ethanediamine; 2 -[2-aminoethyl]-2H-pyrido[3',2'.5,6]thiopyrano[4, 3,2-cd]indazol-5-y]- N,N-dimethyl-1 ,2-ethanediamine; N'-[2-[2-(2-hydroxyethylam ino)ethyl]-2H-pyrido[3',2 5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; 2 2 -(dimethylamino)ethyl].2H..pyrido[3' 2: S,6]thiopyrano[4, 3,2cd]indazo-5-y]-1 ,2-ethanediamine, 'H N.M.R. (200 MHz) in D 2 0 8.69 ppm 1H); 8.49 ppm J=5.7 Hz, 1H); 8.22 ppm J=5.7 Hz, 1 7.39 ppm J=8.7 Hz, 1 7.27 ppm J=8.5 Hz, 1H); 4.90 ppm J=5.8 Hz, 2H); 3.91 ppm J=5.8 Hz, 2H); 3.63 ppm J=5.8 Hz, 2H); 3.35 ppm J=5.8 Hz, 2H); 3.0 ppm 6H); WO 97/02267 WO 9702267PCTIUS96/10966 24 N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4,3,2-cdlindazol- 5-yI]-1 ,2-ethanediamine; N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3' ,2':5,6]thiopyrano[4, 3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; N'-[2-[2-aminoethyl]-2H-pyrido[3',2': 5,6lthiopyrano(4, 3,2-cdindazol-5-y].1 ,2ethanediamine; 2 -[2-(2-hydroxyethylamino)ethy].2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2cd]indazol-5-y]-1 ,2-ethanediamine; 2 2 -(dimethyamino)ethyI]-2H-pyrido[3',2:56]thiopyrano[ 4 3 2 cdlindazol-5-yl]-N-methyl-1 ,2-ethanediamine, m.p. 114-115 0 C, 'H N.M.R. in CDCI 3 8.27 ppm (dd, J=4.7 Hz, J=7.8 Hz, IH); 8.14 ppm (dd, J=1.8 Hz, J=7.8 Hz, 1 7.08 ppm (dd, J=4.7 Hz, J=7.8 Hz, 1IH); 6.89 ppm J=8.7 Hz, 1IH); 6.88 ppm J=8.7 Hz, 1 4.34 ppm J=7.1 Hz, 2H); 3.30 ppm J=5.7 Hz, 2H); 2.86 ppm J=5.7 Hz, 2H); 2.80 ppm J=7.1 Hz, 2H); 2.47 ppm 3H);2.30 ppm 6H); (11) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; m.p. 120-124*C, 'H N.M.R. in CDC1 3 9.07 ppm 1 8.23 ppm J=5.4 Hz, 1 7.11 ppm J=5.4 Hz, 1 6.93 ppm J=8.8 Hz, 1 6.86 ppm J=8.8 2o Hz, 1IH); 5.42 ppm (br s,I1H); 4.36 ppm J=7.1 Hz, 2H); 3.36 ppm J=5.6 Hz, 2H); 2.96 ppm J=7.1 Hz, 2H); 2.93 ppm J=5.5 Hz, 2H); 2.62 ppm J=7.1 Hz, 4H); 2.59 ppm 3H); 1.02 ppm J=7. 1, Hz 6 H); (12) 2 -[2-(methylamno)ethyl]2H-pyrido[3',2':5,6]thiopyrano[4,3,2cd]indazol-5-yI]-N-methyl-1 ,2-ethanediamine; (13) N'-[2-[2-aminoethyl]-2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2-cdlindazol-5-y]-Nmethyl-i ,2-ethanediamine; (14) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2cd]indazol-5-yI]-N-methyl-1 ,2-ethanediamine; N'[-2(iehlmn~ty]2-prd['2:,]hoyao432 cdjindazol-5-yI-N-hydroxyethyl..l,2-ethanediamine; (16) 2 -[2-(diethylamino)ethyl]2H-pyrido[3',2':5,6]thiopyrano[4,3,2-cd]indazolhydroxyethyl -1 ,2-ethanediamine; WO 97/02267 WO 9702267PCTIUS96/10966 (17) N'-[2-[2-(methylamino)ethyl]-2 H-pyrido[3,2': 5,6]thiopyrano[4, 3,2-cd]indazolhydroxyethyl -1 ,2-ethanediamine; (18) N'-[2-[2-aminoethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cdlindazol-5.yI]-N hydroxyethyl -1 ,2-ethanediamine; (19) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2hydroxyethyl -1 ,2-ethanediamine; N-[2-[2-(dimethylamino)ethyl]-2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2cd]indazol-5-yI]-2-aminoethanol; (21) N-[2-[2-(diethylamino)ethyl]-2 H-pyridof 3' 5,6lthiopyrano[4, 3,2-cd]indazol- 5-yI]-2-aminoethanol; (22) N-[2-[2-(methylamino)ethyll-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-2-aminoethanol; (23) N-[2-[2-aminoethyl]-2 H-pyrido[3',2': 5,6]thiopyrano[4, 3,2-cd]indazol-5-yI]- 2aminoethanol; (24) N-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3',2': 5,6]thiopyrano(4, 3,2- 2-aminoethanol; N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4,3':5,6]thiopyrano[4,3,2- N-dimethyl-1 ,2-ethanediamine; (26) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[4',3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N, N-dimethyl-1,2-ethanediamine; (27) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4', 3':5,6]thiopyrano[4,3,2-cd]indazo- N-dimethyl-1 ,2-ethanediamine; (28) N'-[2-[2-aminoethyl]- 2H-pyrido[4', 5,6]thiopyrano(4,3,2-cd]indazol-5-y]- N,N-dimethyl-1 ,2-ethanediamine; (29) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2cd]indazo-5-yIJ-1 ,2-ethanediamine; (31) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-1 ,2-ethanediamine; (32) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4',3': 5,6]thiopyrano[4,3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; WO 97/02267 WO 9702267PCT/US96/10966 26 (33) N'-[2-[2-aminoethy]-2H-pyrido[4,3': 5,6]thiopyrano[4, 3,2-cd]indazol-5-yl]- I ,2-ethanediamine; (34) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2cd]indazol-5-yI]-1 ,2-ethanediamine; (35) N'-[2-[2-(dimethylamino)ethyl]-2 H-pyrido[4', 5,6]thiopyrano[4, 3,2cd]indazo-5-yl]-N-methyl-1 ,2-ethanediamine; (36) N'[-2(ithlmn ty]-Hprd 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (37) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cdlindazol- 5-yl]-N-methyl-1 ,2-ethanediamine; (38) N'-[2-[2-aminoethyl]-2 H-pyrido[4',3': 5,6]thiopyrano[4, 3,2-cd]indazol-5-yl]-Nmethyl-i ,2-ethanediamine; (39) N'-12-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2cd]indazol-5-y]-N-methyl-1 ,2-ethanediamine; (40) N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4', 3':5,6]thiopyrano[4, 3,2cd]indazo-5-yI]-N-hydroxyethyl-1 ,2-ethanediamine; (41) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd]indazolhydroxyethyl -1 ,2-ethanediamine; (42) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4,3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-N- hydroxyethyl -1 ,2-ethanediamine; (43) N'-[2-[2-aminoethyl]-2H-pyrido[4,3': 5,6]thiopyrano[4, 3,2-cdlindazol-5-y]-Nhydroxyethyl -1 ,2-ethanediamine; (44) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4',3': 5,6]thiopyrano[4, 3,2hydroxyethyl -1 ,2-ethanediamine; (45) N-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2cd]indazol-5-yI]-2-aminoethanol; (46) N-[2-[2-(diethylamino)ethyl]-2H-pyrido[4', 3:5,6]thiopyrano[4, 3,2-cdjindazol- 5-yI]-2-aminoethanol; (47) N-[2-[2-(methylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-2-aminoethanol; (48) N-[2-[2-aminoethyl]-2H-pyrido[4', 5,6]thiopyrano[4,3,2-cd]indazol-5-yl]- 2aminoethanol; WO 97/02267 WO 9702267PCTIUS96/10966 27 (49) N-[2-[2-(2-hydroxyethylamino)ethyl]..2H-pyrido[4' 5,6]thiopyrano[4 3,2- 2-aminoethanol; N'-[2-[2-(dimethylamino)ethy]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2cd]indazol-5-yl]-N,N-dimethyl-1 ,2-ethanediamine, 'H N.M.R. (200 MHz) in CDC1 3 9.09 ppm 1 8.25 ppm J=5.3 Hz, 1IH); 7.15 ppm J=5.4 Hz, 1IH); 6.99 ppm J=8.8 Hz, 1IH); 6.90 ppm J=8.8 Hz, 1 4.45 ppm J=6.9 Hz, 2H); 3.34 ppm J=5.9 Hz, 2H); 3.00 ppm J=6.9 Hz, 2H); 2.87 ppm J=5.9 Hz, 2H); 2.45 ppm 6H); 3.29 ppm 6H); (51) N'-[2-[2-(diethylamino)ethyl]-2H..pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N,N-dimethyl-1 ,2-ethanediamine, 1 H N.M.R. (200 MHz) in CDC1 3 8.29 ppm (dd, J=1.8 Hz, J=3.8 Hz, IH); 8.16 ppm (dd, J=1.8 Hz, J=7.8 Hz, 1H); 7.10 ppm (dd, J=4.7 Hz, J=7.3 Hz, 1H); 6.97 ppm J=8.8 Hz, 1 6.89 ppm J=8.8 Hz, I 4.41 ppm J=7.1 Hz, 2H); 3.33 ppm J=6.1 Hz, 2H); 3.04 ppm J=7.1 Hz, 2H); 2.71 ppm (mn, 6H); 2.33 ppm 6H); 1.05 ppm 6H).; (52) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N,N-dimethyl-1 ,2-ethanediamine; (53) N'-[2-[2-aminoethyll- 2H-pyrido[3',4':5,6lthiopyrano[4, 3,2-cd]indazol-5-yI]- N,N-dimethyl-1 ,2-ethanediamine; (54) N'-[2-[2-(2-hydroxyethylamno)ethyll2H-pyrido[3',4': 5, 6lthiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; N'-[2-[2-(diethylamino)ethyI]-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-1 ,2-ethanediamine; (56) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3',4' :5,6]thiopyrano[4,3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; (57) N'-[2-[2-aminoethyll-2H-pyrido[3',4':

S,

6 ]thiopyrano[4,3,2-cd]indazol-5-yl] 1 ,2-ethanediamine; (58) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2cd]indazol-5-y]-1 ,2-ethanediamine; (59) N'[-2(iehlmnoehl Hprd['4': 5,6]thiopyrano[4,3,2cd]indazo-5-yi]-N-methyl.1 ,2-ethanediamine; N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; ii in ~rrra WO 97/02267 PCTIUS96/10966 28 (61) N'-[2-[2-(methylamino)ethyl].2H-pyrido[3',4' :5,6]thiopyrano[4, 3 ,2-cd]indazol- 5-yl]-N-methyl-1 ,2-ethanediamine; (62) N'-[2-[2-aminoethyl]-2H-pyrido[3',4':5,6]thiopyrano[4, 3 methyl-I ,2-ethanediamine; (63) 2 2 -(2-hydroxyethylamino)ethyl]2H.pyrido[3 5,6]thiopyrano[4, 3,2cd]ifldazol-5-yl]-N-methyl-1 ,2-ethanediamine; (64) N'-[2-[2-(dimethylamino)ethyl]2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2cd]indazol-5-yl]-N-hydroxyethyl-1I,2-ethanediamine; N'-[2-[2-(diethylamino)ethyl]2H-pyrido[3',4': 5, 6]thiopyrano[4, 3,2-cdjindazol- 5-yl]-N- hydroxyethyl -1 ,2-ethanediamine; (66) N'-[2-[2-(methylam ino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4,3,2-cd]indazolhydroxyethyl 2-ethanediamine; (67) N'-[2-[2-aminoethyl]-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2-cd]indazol-5-yl]-Nhydroxyethyl -1 ,2-ethanediamine; (68) 2 2 -(2-hydroxyethylamino)ethy].2H.pyrido[3' 5, 6]thiopyrano[4, 3,2hydroxyethyl -1 ,2-ethanediamine; (69) N-[ 2 -[2-(dimethylamino)ethyl]2H-pyrido[3' 5,6]thiopyrano[4,3,2cd]indazol-5-yI]-2-aminoethanol; N-[2-[2-(diethylamino)ethyl]-2H-pyrido[3',4'. 5,6]thiopyrano(4, 3,2-cd]indazol- 5-yi]-2-aminoethanol; (71) N-[ 2 2 -(methylamino)ethyl]..2H..pyrido[3 4:5,6thiopyrano[4,3,2-cd]indazol- 5-yI]-2-aminoethanol; (72) N-[2-[2-aminoethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol-5-yi]- 2aminoethanol; (73) N-[ 2 -[2-(2-hydroxyethylamino)ethyl-2 H-pyrido[3',4':5, 6]thiopyrano[4, 3,2- 2-aminoethanol; (74) 2 2 (dimethyamino)ethyl]-2H-pyrido[21 3 ':56]thiopyrano[4,3,2cd]ifldazol-5-yl]-N,N-dimethyl.1 ,2-ethanediamine; 2 -[2-(diethylamino)ethyl].2H..pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-N,N-dimethyl-1 ,2-ethanediamine; (76) 2 -[2-(methylamino)ethyl]-2H-pyrido12',3': 5,6]thiopyrano[4,3,2-cd]indazol- 5-yI]-N,N-dimethyl-1 ,2-ethanediamine; WO 97/02267 WO 9702267PCTIUS96/10966 29 (77) N'-[2-[2-aminoethyl]- 2 H-pyrido[2', 5,6]thiopyrano[4, 3,2-cdjindazoI5-y] N,N-dimethyl-1 ,2-ethanediamine; (78) 2 2 2 -hydroxyethylamino)ethy]-2H-pyrido[2',3':5,6]thiopyrano[4,3,2- N-dimethyl-1 ,2-ethanediamine; (79) 2 2 -(dimethylamino)ethyl]2H-pyrido[2,3': 5, 6]thiopyrano[4, 3,2cd]indazo-5-yl]-1 ,2-ethanediamine; 2 -[2-(diethylamino)ethyl]-2H-pyrido[2',3': 5,6]thiopyrano[4,3,2-cd]indazol.

5-yI]-1 ,2-ethanediamine; (81) 2 -[2-(methylamino)ethyl]-2H-pyrido[2' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-1 ,2-ethanedjamine; (82) N'-[2-[2-aminoethyl]-2H-pyrido[2', 5,6]thiopyrano[4, 3 1 ,2-ethanediamine; (83) 2 2 2 -hydroxyethylamino)ethy].2H..pyrido 2 5,6]thiopyrano[4,3,2cdlindazol--y]-1 ,2-ethanediamine; (84) 2 -[2-(dimethylamino)ethyl]-2H.pyrido[2' 5,6]thiopyrano[4, 3,2cdlindazo1-5-y]-N-methyl.1 ,2-ethanediamine; 2 -[2-(diethylamino)ethyl]-2H.pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (86) 2 -[2-(methylamino)ethyl]-2H-pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (87) N'-[2-[2-aminoethyl]-2H-pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazol-5-yi].Nmethyl-I ,2-ethanediamine; (88) 2 2 2 -hydroxyethylamino)ethy].2H.pyrido[2' 5,6]thiopyrano[4, 3,2cd]indazol-5-yI]-N-methy-1 ,2-ethanediamine; (89) 2 2 (dimethylamino)ethy].2Hpyrido[23:,6hiopyrano[ 4 3 2 cd]indazo-5-yl]-N-hydroxyethyl-1 ,2-ethanediamine; 2 -[2-(diethylamino)ethyl]-2H-pyrido[2',3': 5,6]thiopyrano(4,3,2-cd]indazolhydroxyethyl -1 ,2-ethanediamine; (91) 2 2 -(methylamino)ethyl]-2H..pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-N- hydroxyethyl -1 ,2-ethanediamine; (92) N'-[2-[2-aminoethyl]-2H-pyrido[2' 3':5,6]thiopyrano[4, 3,2-cd]indazol-5-yl]-Nhydroxyethyl -1 ,2-ethanediamine; WO 97/02267 PCTIUS96/10966 (93) 2 2 2 -hydroxyethylamino)ethyl].2H-pyrido[2',3': 5,6]thiopyrano[4 3,2hydroxyethyl -1 ,2-ethanediamine; (94) N-[ 2 -[2-(dimethylamino)ethyl]-2H..pyrido[2', 5,6]thiopyrano[4, 3,2cd]indazol-5-yl]-2-aminoethanol; (95) N-[2-[2-(diethylamino)ethyl]-2H..pyrido[2', 5,6]thiopyrano[4,3,2-cdjindazol- 5-yl]-2-aminoethanol; (96) N-[ 2 -[2-(methylamino)ethyl]2H-pyrido[2' ,3':5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-2-aminoethanol; (97) N-[2-[2-aminoethyl]-2H-pyrido[2', 5,6]thiopyrano[4, 3 ,2-cd]indazol-5-yJ- 2aminoethanol; (98) N-[ 2 2 2 -hydroxyethylamino)ethyl..2H..pyrido[2',3': 5,6]thiopyrano[4, 3,2- 2-aminoethanol.

Example 10 comroundsADI, D -NHCO-CH-NR,.R.4 Starting from a chloroform solution of the suitable compounds prepared according to preparations 7 or 8, by removal of the N-BOC protecting group with dry gaseous hydrogen chloride, the following compounds as hydrochloride salts are obtained: pyrido[3',2':5,6]thiopyrano [4,3,2-cd]indazole, 'H N.M.R. (200 MHz) in D 2 0 3.0 ppm 6H); 3.81 ppm 2H); 4.13 ppm (t, 2H); 4.80 ppm 2H); 7.30 ppm (in, 3H); 8.30 ppm (in, 2H); (2_'[-2dmtyaioehy]5( aioaeyaio-Hpyrido[3',4':5,6]thiopyrano [4,3,2-cd]indazole; pyrido[4',3':5,6]thiopyrano [4,3,2-cd]indazole; pyrido[2',3': 5,6]thiopyrano [4,3,2-cdjindazole; 2 2 -di ethyl am ino)ethy]s5(2'-am ino.acetylamino)-2

H-

pyrido[3',2':5,6]thiopyrano [4,3,2-cd]indazole; (6_'[-2dehlmn~ty]5('aioaeyaio-H pyrido[3' 5,6]thiopyrano 3 ,2-cd]indazole; pyrido[4', 3':5,6]thiopyrano 4 ,3,2-cd]indazole; WO 97/02267 WO 9702267PCTIUS96/10966 31 2 2 -diethylamino)ethyl]-5-(2'-amno-acetylamino)2H-.

pyrido[2', 5,6]thiopyrano 3,2-cd]indazole; 2 2 -methyamino)ethyl]5(2'aminoacetylamino)-2H.

pyrido[3',2': 5,6]thiopyrano 3,2-cd]indazole; (1 2 2 -methylamino)ethyl]-5(2'amino-acetylamino)-2H.

pyrido[3' 5,6]thiopyrano 3,2-cd~indazole; (11) 2 -(2-methylamino)ethyl]-5-(2-aminoacetylamino)-2Hpyrido[4', 5,6]thiopyrano 3,2-cd]indazole; (12) 2 -(2-methylamino)ethyl]-5-(2'-aminoacetylamino)-2H.

pyrido[2',3':5,6]thiopyrano [4,3,2-cd]indazole; (13) 2 -aminoethyl]-5-(2'-aminoacetylamino)-2H-pyrido[3' 5,6]thiopyrano [4,3,2-cd]indazole; (1 4 )N'-[2-aminoethyl]-5-(2'-aminoacetylamino)-2H-pyrido[3' 5,6]thiopyrano [4,3,2-cd]indazole; (15) 2 -aminoethyl]-5(2'-aminoacetylamino)-2H-pyrido[4',3':5,6]thiopyrano 3,2-cd]indazole; (16) N'-[2-aminoethyl]-5-(2'-amino-acetyamino)-2H-pyrido[2' 5,6]thiopyrano 3,2-cd] indazole; (17) 2 2 -hydroxyethylamino)ethyl]5(2'..amino-acetylamino)-2Hpyrido[3',2': 5,6] thiopyrano [4,3,2-cd]indazole; (11 hydroxyethylamino)ethyl]-5(2'-amino-acetylamino)2Hpyrido[3',4':5,6 thiopyrano [4,3,2-cd]indazole; (19) hydroxyethylamino)ethy]5(2'aminoacetylamino)-2Hpyrido[4',3':5,6] thiopyrano [4,3,2-cd]indazole; (20) hydroxyethylamino)ethyl]-5-(2'amino-acetylamino)-2H.

pyrido[2',3':5,6] thiopyrano 3,2-cd]indazole.

ExamplIe 11 compounds D nitro According to the procedures described in examples 1, 3 or 4, starting from the suitable intermediates, the following nitro derivatives are obtained: N,N-ity 5nto2-prd['4': 5,6]th-iopyrano[4, 3,2-cd]indazole-2ethanamine, 1H N.M.R. (200 MHz) in CDCI 3 9.38 ppm 8.50 ppm J=5.4 Hz, 1H); 8.23 ppm J=9.3 Hz, 1 7.45 ppm J=5.4 Hz, 1 7.11 ppm J=9.3 Hz, WO 97/02267 WO 9702267PCT/US96/10966 32 lh); 4.45 ppm J=6.4 Hz, 2H); 2.98 ppm J=6.4 Hz, 2H); 2.54 ppm J=7.1 Hz, 4H); 0.92 ppm J=7.1 Hz, 6H); N, N-diethyl-5-nitro-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine, m.p. 132-133 0

C,

'H N.M.R. (200 MHz) in CDC1 3 8.56 ppm (dd, J=1.7 Hz, J=4.6 Hz, 1 8.43 ppm (dd, J=1.7 Hz, J=7.8 Hz, 1 8.25 ppm J=9.3 Hz, 1IH); 7.33 ppm (dd, J=4.6 Hz, J=7.8 Hz, 1 7.08 ppm J=9.3 Hz, 1 4.44 ppm J=6.4 Hz, 2H); 2.98 ppm J=6.4 Hz, "h9; 2.54 ppm J=7.1 Hz, 4H); 0.94 ppm J=7.1 Hz, 6H); N, N-dimethyl-5-nitro-2H-pyrido[4', 3':5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N, N-diethyl-5-nitro-2H-pyrido[4',3':5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N, N-dimethyl-5-nitro-2H-pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N, N-diethyl-5-nitro-2H-pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N-methyl-5-nitro-2H-pyrido[3' :5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N-ehl5nto2-yio3,'5,]hoyao432c~naoe2 ethanamine; N-methyl-5-nitro-2H-pyrido[4',3':5,6]thiopyrano[4,3,2-cdlindazole-2ethanamine; N-methyl-5-nitro-2H-pyrido[2',3':5,e]thiopyrano[4,3,2-cd]indazole-2ethanamine; (11) 5-nitro-2H-pyrido[3'

S,

6 ]thiopyrano[4,3,2-cd]indazole-2-ethanamine; (12) 5-nitro-2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2-cd]indazole-2-ethanamine; (13) 5-nitro-2H-pyrido[4', 3':5,6]thiopyrano[4, 3,2-cd]indazole-2-ethanamine; (14) 5-nitro-2H-pyrido[2',3':5,6]thiopyrano[4, 3,2-cd]indazole-2-ethanamine; (15) N-hydroxyethyl-5-nitro-2H-pyrido[3' 6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; (16) N-yrxehl--ir-Hpyio3, 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; WO 97/02267 WO 9702267PCTIUS96/10966 33 (17) N-hydroxyethyl-5-nitro-2H-pyrido[4', 5, 6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; (18) N-hydroxyethyl-5-nitro-2H-pyrido[2',3':5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine.

Example 12 comounds D amino According to the procedure described in examples 2 or 5, starting from the nitro derivatives of example 11, the following amino derivatives are obtained: N, N-diethyl-5-amino-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine, 'H N.M.R. (200 MHz) in CDCI 3 9.09 ppm 1 8.24 ppm J=5.5 Hz, 1IH); 7.13 ppm J=5.5 Hz, 1IH); 6.88 ppm J=8.6 Hz, I 6.77 ppm J=8.6 Hz, 1H); 4.31 ppm J=7.9 Hz, 2H); 3.33 ppm (br s, 2H); 2.92 ppm J=7.0 Hz, 2H); 2.58 ppm J=7.1 Hz, 4H); 1.01 ppm J=7.1 Hz, 6H); N, N-diethyl-5-amino-2H-pyrido[3',2': 5,6]thiopyrano[4,3,2-cd]indazole-2ethanamine; N, N-dimethyl-5-amino-2H-pyrido[4',3':5,6]thiopyrano[432cd]indazole-2 ethanamine; N, N-diethyl-5-amino-2H-pyrido[4', 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N, N-dimethyl-5-amino-2H-pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N, N-diethyl-5-amino-2H-pyrido[2',3':5,6]thiopyrano[4,3,2-cd]indazole-2ethanamine; N-methyl-5-amino-2H-pyrido[3' :5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N-methyl-5-amino-2H-pyrido[3' :5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; N-methyl-5-amino-2H-pyrido(4', 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; (10) N-methyl-5-amino-2H-pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazole-2ethanamine; (11) 5-amino-2H-pyrido[3' 5,6]thiopyrano[4, 3, 2-cd]indazole-2-ethanamine; (12) 5-amino-2H-pyrido[3',2':

S,

6 ]thiopyrano[4,3,2-cd]indazole-2-ethanamine; WO 97/02267 PCTIUS96/10%6 34 (13) 5-amino-2H-pyrido[4',3': 5,6]thiopyrano[4, 3 2 -Cd]indazole-2-ethanamine; (14) 5-amino-2H-pyrido[2', 5,6]thiopyrano[4, 3 2 -cd]indazole-2-ethanamine; N-hydroxyethyl-5-amino2H-pyrido[3' ,4':5,6]thiopyrano[4, 3 ,2-cd]indazole-2ethanamine; (16) N-hydroxyethyl-5-amino-2H-pyrido[3',2':5,6]thiopyrano[4, 3,2-cd]indazole-2-.

ethanamine; (17) Nhdoyty--mn-Hprd['3': 5,6]thiopyrano[4, 3 ,2-cd]indazole-2ethanamine; (18) N-hydroxyethy-5-amino.2H-.pyrido[2',3': 5,6]thiopyrano[4, 3,2-cd]indazole-2 ethanamine.

WO 97/02267 PCT/US96/10966 TABLE 11

IC,

0 (rig/mI) Structure Example L1210 SI80 LoVo LoVofDx

N-

i XHC.H20 7 0.0125 0.058 0.027 16.8 N S N"2

N.

IS 4 6 0.008 0.028 2.4 2.6 N Sk

NMON

Nj .x Cd-H2O ex. I0-cpd 1 0.03 0.22 n.d. n.d.

C-1O.MH2O ex. 9-cpd 30 0.014 0.058 n.d. n.d.

HHNH,

.XN2O ex. I0-cpd 2 n.d n.d. 0.04 0.21 HNrH, 0 .HOzH 2 0 8 0.0016 0.0029 0.008 0.26

HN,,-NH,

0.0030 0.0066 0.016 0.027

NH,

ex. 12-cpd 1 0.0015 0.0031 0.036 0.094

NH,

WO 97/02267 WO 9702267PCT1US96/10966 TABLE I1 continue*I Structure Example

IC,

0 (ig/miJ) 801_LoVo L1210 S1 L1I 1 I Lo o/x CI-941 nd doxonzbicin n.d.

0.053 44.5 n.d. 0.58 53.8 n.d. 0.024 0.67 mitoxantrone n.d.

n.d. =not determined

IC,

0 is the concentration of the drug which causes a 50% inhibition of the tumor cells growth CYTOTOXICIWY ON LOVO AND LOVOIDX WAS DETERMINED BY MTT ASSAY FOLLOWING I HOUR OF DRUG EXPOSURE WO 97/02267 WO 9702267PCT/US96/10966 TABLE III (lig/ml) Structure Example LI1210 SI180 LoVo LoVo/Dx ex. I I-cpd 1 0.00061 0.0031 n.d. n.d.

NO,

4 0.00062 0.0032 n.d. n.d.

Na, 43 0.05 0.14 n.d. n.d.

ex. I I1-cpd 2 0.2 0.22 nd. n.d.

NOS

10.08 0.17 n.d. n.d.

NOIC

n.d. not determined ICso is the concentration of the drug which causes a 50% inhibition of the tumor cells growth CYTOTOXICITY ON LOVO AND LOVO/DX WAS DETERMINED BY MTT ASSAY FOLLOWING 1 HOUR OF DRUG EXPOSURE

Claims (5)

1. A compound of formula D Y_ X S N--N B wherein: one of X, Y, Z or T is nitrogen and the others are CH; D is selected from the group consisting of nitro or -NH-A, wherein A is selected 10 in the group consisting of hydrogen, -CO-CH 2 -NR 2 R 3 C1-Clo alkyl; C2-Co alkyl O: having one or two substituents selected from the group consisting of OR 1 and NR 2 R 3 C2-C10 alkyl interrupted by one or two oxygen atoms or by one -NR 4 *.:group, and said C2-Co0 alkyl is optionally substituted by one or two hydroxy (OH) or -NR 2 R 3 groups; I 15 B is selected from the group consisting of Ci-Clo alkyl; C2-C10 alkyl having one or two substituents selected from the group consisting of OR, and -NR 2 R 3 and SC2-C10 alkyl interrupted by one or two oxygen atoms or by one -NR 4 group, wherein said C2-C10 alkyl is optionally substituted by one or two hydroxy (OH) or -NR 2 R 3 groups; R, is selected from the group consisting of hydrogen, Ci-C6 alkyl, -S(0 2 )R 5 and C2-C6 alkyl optionally substituted by -NR 2 R 3 R 2 and R 3 may be the same or different and are selected from the group consisting of hydrogen, C 1 -Clo alkyl, and C2-C10 alkyl substituted with one or two hydroxy (OH) groups; or R 2 and R 3 taken together with the nitrogen atom to which they are linked form a 5- or 6-member aromatic or non-aromatic heterocyclic ring which optionally contains another heteroatom such as sulfur, oxygen or nitrogen; R 4 is selected from the group consisting of hydrogen, C-Clo alkyl, C2-C10 z /r hydroxyalkyl, and C2-C10 alkyl substituted with -NR 2 R 3 39 R 5 is selected from the group consisting of 0 i-CIO alkyl, phenyl, and phenylalkyl; as free bases and their salts with pharmaceutically acceptable acids.

2. The compound of claim 1 wherein Cj-CjO alkyl is selected from the group consisting of methyl, ethyl, n-propyl, sec-propyl, n-butyl, sec-butyl, tert-butyl, n- pentyl, n-hexyl.

3. The compound of claim 1 or claim 2 wherein phenylalkyl is 4-methylphenyl.

4. The compound of any one of claims I to 3 wherein the -NR 2 R 3 substituent is selected from the group consisting of 1-imidazolyl, 4-hydroxy-1-imidazolyl, 2-imino- I (3H)-imidazolyl, I -pyrrolyl, I -tetra hyd ropyrrolyl, 1 -pyrazolyl, 4-morpholinyl, 1- piperidinyl, 1-piperazinyl, I-(4-methyl) piperazinyl, and I-(4-benzyl) piperazinyl. The compound of claim 1 selected from the group consisting of N, N-dimethyl-5-nitro-2H-pyrido[3',2':5, 6]thiopyrano[4, 3,2-cdlindazole-2. ethanamine; (2 tyamio th l]2H p rd 3,2:56thip rn -d d z l 15 S-12ehaeiai

155-yl]-1 ,2-ethanediamine; 2 2 -(mithnoethyl] th-yrid].2yrid[3D2iopyratho4yra-dno,,daz15]ido S-l 2ethanediamine; 2 N'-[2-[2-(2hdytyaminoethyl]-2 H-pyrido3',2': 5,6]thiopyrano[4, 3,2-dinao-yll,2 cdidzl5y]12ethanediamine; 2 2 2 hyroythlmneylmio2eh-yrido3.2H.. ]hpyrian.6]hopyrf3[ 4 3 cd]indazol-5-y]-1,-ethyan,edianeimie iethylam in o)ethyl ]2H pyri do[3,2': 56]th iopyrano 3,2- inaoI cdinzol5-yl]-N-methy ,2-ethanediamine 2 2amiethlaino)et-H-yrid['2pyridhopyr2an,6thopyrn43,2-cdjindazol-N 5ymethyl-1 ,2-ethanediamine 30(8) 2 2 -(mehylamoxytyno)e.h.ydl]2': 56tidopyran5,6toy32-andazol,- c-yl]-N-methyli1-N2-ethnediamithne imie (91) N'-[2-[2-admeto ino)e tyrd]s'2': S,6] ',:,6thiopyrano[4 cd]indazol-5-yl]-N-mhyl-1 th-,2-ethanediamine L ull WO 97/02267 WO 9702267PCTIUS96/10966 (12) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3',2': 5,6]thiopyrano[4, 3,2-cd]indazol- hydroxyethyl -1 ,2-ethanediamine; (13) N'[-2(ehlmn~eh -Hprd['25,6]thiopyrano[4, 3,2-cd]indazol- hydroxyethyl -1 ,2-ethanediamine; (14) N'-[2-[2-aminoethyl]-2H-pyrido[3' ,2':5,6]thiopyrano[4, 3,2-cdindazo-5yl]-N hydroxyethyl -1 ,2-ethanediamine; 2 -[2-(2-hydroxyethylamino)ethy].2H-pyrido[3' 5,6]thiopyrano[4 3,2- hydroxyethyl -1 ,2-ethanediamine; (16) N-[2-[2-(dimethylamino)ethyl]-2H..pyrido[3' :5,6]thiopyrano[4, 3,2- cd]indazol-5-yI]-2-aminoethanol; (17) N-[2-[2(diethyamino)ethy]2H-pyridof 3',21: ,6]thiopyrano[4,3,2-cd]indazol- 5-yi]-2-aminoethanol; (18) N-[2-[2-(methylamino)ethyI]-2H-pyrido[3',2'; 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-2-aminoethanol; (19) N-[2-[2-aminoethyI]-2H-pyrido[3I 21: 5,6]thiopyrano[4, 3,2-cd]indazol-5-y]- 2- aminoethanol; N-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3' ,2':5,6]thiopyrano[4, 3,2- 2-aminoethanol; (21) N'-[2-[2-(dimethylamino)ethyl].2H..pyrido[4 5,6]thiopyrano[4, 3,2- cd]indazol-5-yl]-N, N-dimethyl-1 ,2-ethanediamine; (22) N'-[2-[2-(diethylamino)ethyl]-2H..pyrido[4',3': 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N,N-dimethyl-1 ,2-ethanediamine; (23) N'-[2-[2-(methylamino)ethyII-2H-pyrido[4',3': 5, 6jthiopyrano[4, 3, 2-cd]indazol- N-dimethyl-1 ,2-ethanediamine; (24) N'-[2-[2-aminoethyl]- 2 H-pyrido[4',3':5,6thiopyrano[432-cd]indazoI-5yI]- N,N-dimethyl-1 ,2-ethanediamine; 2 -2-(2-hydroxyethylamino)ethyl].2H..pyrido[4',3':5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; (26) N-2[-dmtyaioehl2Hprd4'3': 5,6]thiopyrano[4, 3,2- cd]ifldazol-5-yI]-1 ,2-ethanediamine; (27) iethyl am ino)ethyI]-2 H pyrido[4l, T: 5,6]th iopyrano[4,3,2-cd] indazol 5-yII-1 ,2-ethanediamine; WO 97/02267 PTU9/06 PCT/US96/10966 41 (28) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; (29) N'-[2-[2-aminoethyl]-2H-pyrido[4' 5,6]thiopyrano[4,3,2-cdlindazo-5-YI]- I ,2-ethanediamine; (30) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2- cd]indazol-5-y]-1 ,2-ethanediamine; (31) N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4', 5,6]thiopyrano[4,3,2- cd]indazo1-5-yl]-N-methyl-1 ,2-ethanediamine; (32) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (33) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4',3:5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (34) N'-[2-2-aminoethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd~indazol-5-yJ-N- methyl-I ,2-ethanediamine; (35) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4', 3':5,6]thiopyrano[4, 3,2- cd]indazol-5-y]-N-methyl-1 ,2-ethanediamine; (36) N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4',3':5,6]thiopyrano[4, 3,2- cd]indazo-5-yI]-N-hydroxyethyl-1 ,2-ethanediamine; (37) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yl]-N- hydroxyethyl -1 ,2-ethanediamine; (38) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2-cd]indazol- hydroxyethyl -1 ,2-ethanediamine; (39) N'-[2-[2-aminoethyl]-2H-pyrido[4',3':5,6]thiopyrano[4,3,2-cd]indazol-5-yl]-N hydroxyethyl -1 ,2-ethanediamine; (40) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2 H-pyrido[4',3': 5,6]thiopyrano[4, 3,2- hydroxyethyl 2-ethanediamine; (41) N-[2-[2-(dimethylamino)ethyl]-2H-pyrido[4',3': 5,6]thiopyrano[4, 3,2- cd]indazol-5-yI]-2-aminoethanol; (42) N-[2-[2-(diethylamino)ethyl]-2H-pyrido[4', 3':5,6]thiopyrano[4,3,2-cd]indazol- 5-yl]-2-aminoethanol; (43) N-[2-[2-(methylamino)ethyI]-2H-pyrido[4' :5,6]thiopyrano[4,3,2-cd]indazol- 5-yI]-2-aminoethanol; WO 97/02267 PTU9106 PCTIUS96/10966 42 (44) N-[2-[2-aminoethyl]-2H-pyrido[4', 5,6]thiopyrano[4, 3,2-cd]indazol-5-y]- 2- aminoethanol; N-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[4' 5,6]thiopyrano[4, 3,2- 2-aminoethanol; (46) N'-[2-[2-(dimethylamino)ethyt]-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine, (47) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2- ,2-ethanediamine, (48) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2- cd]indazol-5-yl]-N, N-dimethyl-1 ,2-ethanediamine; (49) N'-[2-[2-aminoethyl]- 2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol-5-ylI- N, N-dimethyl-1 ,2-ethanediamine; N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3', 5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; (51) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3' ,4':5,6lthiopyrano[4,3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; (52) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yi]-1 ,2-ethanediamine; (53) N'-[2-[2-aminoethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3, 1 ,2-ethanediamine; (54) N'-[2-[2-(2-hydroxyethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2- cd]indazol-5-y]-1 ,2-ethanediamine; N'-[2-[2-(dimethylamino)ethyl]-2H-pyrido[3' ,4':5,6]thiopyrano[4,3,2- cd]indazo-5-yl]-N-methyl-1 ,2-ethanediamine; (56) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2-cdlindazol- 5-yI]-N-methyl-1 ,2-ethanediamine; (57) N'-[2-[2-(methylamino)ethyl]-2H-pyrido[3',4': 5,6]thiopyrano[4,3,2-cd]indazol- 5-yl]-N-methyl-1 ,2-ethanediamine; (58) N'-[2-[2-aminoethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cdjindazol-5-y]-N- methyl-i ,2-ethanediamine; (59) N'-[2-[2-(2-hydroxyethylamino)ethyl-2FL-pyridoE3' 5,6]thiopyrano[4, 3,2- cd]indazol-5-yI]-N-methyl-1 ,2-ethanediamine; WO 97/02267 PCTIUS96/10966 43 2 -[2-(dimethylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2- cd]indazol-5-yI]-N-hydroxyethyl-1,2-ethanediamine; (61) N'-[2-[2(diethyamino)ethyl]-2H-pyrido[33 4':5,6]thiopyrano[4, 3,2-cd]indazol- hydroxyethyl -1 ,2-ethanediamine; (62) 2 -[2-(methylamino)ethyl]-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2-cd]indazo. hydroxyethyl -1 ,2-ethanediamine; (63) N'[-2ainehl Hpyio3,'5,6]thiopyrano[4, 3,2-cd]indazol.5yi].N hydroxyethyl -1 ,2-ethanediamine; (64) 2 2 -(2-hydroxyethylamino)ethyl].2H-pyrido[3 5,6]thiopyrano[4, 3,2- cd]indazol-5-y]-N- hydroxyethyl -1 ,2-ethanediamine; N-[2-[2-(dimethylamino)ethyl]2H-pyrido[3',4', 5,6]thiopyrano[4, 3,2- cdlindazol-5-yl-2-aminoethanol; (66) N-[2-[2-(diethylamino)ethyl]-2H-pyrido[3 5, 6]thiopyrano(4, 3,2-cdjindazol- 5-yI]-2-aminoethanol; (67) N-[ 2 -[2-(methylamino)ethyl]-2H-pyrido[3' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yi]-2-aminoethanol; (68) N-[2-[2-aminoethyl]-2H-pyrido[3' ,4':5,6]thiopyrano[4, 3,2-cd]indazol-5-yI]- 2- aminoethanol; (69) N-[ 2 2 -(2-hydroxyethylamino)ethyl]2H.pyrido[3 5,6]thiopyrano[4, 3,2- cdlindazol-5-yI]- 2-aminoethanol; 2 -[2-(dimethylamino)ethyl]-2H-pyrido[2',3': 5,6]thiopyrano[4, 3,2- N-dimethyl-1 ,2-ethanediamine; (71) N'-[2-[2-(diethylamino)ethyl]..2H-pyrido[2' 3':5,6]thiopyrano[4, 3,2-cd]indazol- N-dimethyl-1 ,2-ethanediamine; (72) N'-[2-[2-(methylamino)ethyl].2H-pyrido[2' 5,6]thiopyrano[4, 3,2-cd]indazol- 5-yI]-N,N-dimethyl-1 ,2-ethanediamine; (73) N'-[2-[2-aminoethyl]- 2 H-pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazol-5-y]- N, N-dimethyl-1 ,2-ethanediamine; (74) 2 2 -(2-hydroxyethylamino)ethy].2H..pyrido[2',3': 5,6]thiopyrano[4, 3,2- cd]indazol-5-y].N, N-dimethyl-1 ,2-ethanediamine; 2 -[2-(dimethylamino)ethyl]-2H-pyrido[2',3', 5,6]thiopyrano[4,3,2- cd]indazo-5-y]-1 ,2-ethanediamine; WO 97/02267 PCTIUS96I1096 44 (76) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[ 2 5,6]thiopyrano[4,3,2-cd]inddzol- ,2-ethanediamine; (77) N'-[2-[2-(methylamino)ethyl]..2H-pyrido[2', 5,6]thiopyrano[4, 3,2-dindaol. ,2-ethanediamine; (78) N'-[2-[2-aminoethyl]-2H-pyrido[2', 5, 6]thiopyrano[4, 3,2-cdjindzoI-5-y] I ,2-ethanediamine; (79) 2 2 2 -hydro)xIethyamino)ethy]2H-pyrido[2 5,6]thiopyrano[4,3,2 cd]indazol-5-yI]-1 1 2-ethanediamine; 2 2 -(dimethylamino)ethyl]..2H-pyrido[2 5,6]thiopyrano[4, 3,2- cd]indazol-5-y]-N-methyl.1 ,2-ethanediamine; (81) 2 -[2-(diethylamino)ethyl]-2H..pyrido[2',3': 56]thiopyrano[4,3,2-cd]indazol- 5-yl]-N-methyl-1 ,2-ethanediamine; (82) 2 -[2-(methylamino)ethy].2H-pyrido[2 5, 6 ]thiopyrano(4,3,2..cdlindazo. 5-yI]-N-methyl-1 ,2-ethanediamine; (83) N'-[2-[2-aminoetbyl]..2H-pyrido[2', 5,6]thiopyrano[4, 3,2-cd]indazol-5-yi]-N methyl-I ,2-ethanediamine; (84) 2 2 -(2-hydroxyethylamino)ethy].2H..pyrido2 5,6]thiopyrano[4, 3,2- cd]indazol-5-yI]-N-methyl-1,2-ethanediamine; 2 -[2-(dimethylamino)ethyl]..2H-pyrido[2 5,6]thiopyrano[4, 3,2- cd]indazol-5-yl]-N..hydroxyethyl-1,2-ethanediamine; (86) N'-[2-[2-(diethylamino)ethyl]-2H-pyrido[2',-3': 5,6]thiopyrano[4,3,2-cd]indazol- hydroxyethyl -1 ,2-ethanediamine; (87) 2 -[2-(methylamino)ethyl].2H.pyrido[2 5,6]thiopyrano[4, 3, 2-cdlindazol- hydroxyethyl -1 ,2-ethanediamine; (88) N'-[2-[2-aminoethyl]2H-pyrido[2',3': 5,6]thiopyrano[4, 3,2-cdlindazol-5-yIJ-N. hydroxyethyl -I ,2-ethanediamine; (89) 2 2 -(2-hydroxyethylamino)ethyl].2H..pyrido[ 2 5,6lthiopyrano[4, 3,2- hydroxyethyl -1 ,2-ethanediamine; N-[ 2 2 -(dimethylamino)ethyl].2H-pyrido[2',3': 5,6]thiopyrano[4 3,2- cd~indazol-5-yI]-2aminoethanol; (91) N-[ 2 2 -(diethylamino)ethyl]-2H-pyrido[2' 3':5,6]thiopyrano[4,3,2-cd]indazol- 5-yl]-2-aminoethano; ii NIiF rllirvrilIuilriii iirinhiniiiiiiiirnwririiii iinTn~] (92) N-[2-[2-(methytamino)ethyl..2H-pyrido[2',3': 5, 6lthiopyrano[4, 3 2 -cd]indazo.. 5-YI]-2-aminoethanol; (93) N-[2-[2-aminoethyl-2H-pyrido[2' 5 6 Ithiopyrano(4, 3 2 -Cdjindazol-5yj.. 2- aminoethanol; (94) N-[ 2 2 -(2-hydroxyethylamino)ethylII2H-pyrido 2 5, 6lthiopyrano[4,3,2- 2-aminoethanol. 6. The compound of claim 1 selected from the group consisting of: 2 2 -dimethylamino)ethyII..5.(2'.amino.acetylamino)- 2 H- pyrido(3',2':5, 6]thiopyrano 3,2-cdlindazole, 2 2 2 -dimethylamino)ethyI1-5(2'aminoacetylamino)- 2 H- pyrido[3',4': 5, 6lthiopyrano 3,2-cd]indazole; 2 2 -dimethylamino)ethyiI..5(2'.aminoacetylamino)- 2 H- pyrido[4', 5,Gjthiopyrano 3 ,2-cdjindazole; 2 2 -dimethylamino)ethyll-5-(2'..amino-acetylamino)- 2 H- i pyrido[2',3':5,6lthiopyrano 4 ,3,2-cd]indazole; 2 2 -diethylamino)ethyll-5..(2'-amino-.acetylamino)- 2 H- pyrido[3',2': 5, 6]thiopyrano 4 ,3,2-cdjindazole; *(6)_N'-[2-(2-diethylamino)ethyI15-.(2'.aminoacetylamino)- 2 H- pyrido[3', 5, 6]thiopyrano 3,2-cd]indazole; 2 2 -diethylamino)ethyI1.5(2'.aminoacetyamino)- 2 H- Dvridor4',3': 5,6lthiopyrano [4,32clnaoe 2 2 -diethylamino)ethyII.5(2'aminoacetylamino)- 2 H- pyrido[2', 5, 6]thiopyrano 3,2-cdjindazole; 2 2 -methylamino)ethyI5.(2'aminoacetylamino)- 2 H- pyrido[3',2': 5, 6lthiopyrano 3,2-cd]indazole; (1 2 2 -methylamino)ethytI.5.(2'.aminoacetylamino)- 2 H- pyrido(3',4':S,6]thiopyrano [4,3,2-cdlindazole; (11) 2 2 -methyam ino) ethyl inoacety am ino)- 2 H pyrido[4', 5, 6]thiopyrano 3 ,2-cd]indazole; (12) 2 2 methylamino)ethyI].s-(2P-aminoacetylamino)- 2 H- pyrido[2' 5, 6]thiopyrano 3,2-cdjindazole; mnety]5('-mn-ctlaio-Hpyio3,'5,6jthiopyrano 46 (1 -amino ethyl ino-acety Iam ino)-2 H-pyri do 5,6]th iopyrano [4,3,2-cdlindazole; N'-[2-aminoethyl]-5-(2'-amino-acetylamino)-2H-pyrido(4', 5,6]thiopyrano [4,3,2-cd]indazole; (16) N'-f2-aminoethyl]-5-(2'-amino-acetylamino)-2H-pyrido[2', 3':5,6]thiopyrano 3,2-cd]indazole; (17) N'-[2-(2-hydroxyethylamino)ethyl]-5-(2'-amino-acetylamino)-2H- pyrido(3',2': 5,6] thiopyrano 3,2-.cd]indazole; (1 hydroxyethylamino)ethyl]-5-(2'-amino-acetylamino)-2H- pyrido[3',4':5,6 thiopyrano, 3,2-cd]indazole; (19) hydroxyethylamino)ethyl]-5-(2'-amino-acetylaminio)-2H- pyrido(4', 5,6] thiopyrano 3,2-cd]indazole; (20) hydroxyethylamino)ethyl]-5-(2'-amino-acetylamino)-2H- pyrido(2',3':5,6] thiopyrano, [4,3,2-cd]indazole. 15 7. The compound of claim 1 selected from the group consisting of: N, N-diethyl-5-nitro-2H-pyrido[3',4': 5,6]thiopyrano[4, 3,2-cd]indazole-2- ethanamine, N, N-diethyl-5-nitro-2H-pyrido[3',2': 5,6lthiopyrano[4, 3,2-cd]indazole-2- ethanamine; N, N-dimethyl-5-nitro-2H-pyrido(4' 5,6]thiopyrano[4,3,2-cd]indazole-2- ethanamine; N, N-.diethyl-5-nitro-2H-pyrido(4', 5,6]thiopyrano[4, 3,2-cd]indazole-2- .:ethanamine; N,N-dimethyl-5-nitro-2H-pyrido(2', 5,6]thiopyrano[4, 3,2.-cd]indazole-2- ethanamine; N, N-diethyl-5-nitro-2H-pyrido[2', 5, 6]thiopyrano[4, 3, 2-cd]indazole-2- ethanamine; N-methyl-5-nitro-2 H-pyridof 5, 6jthiopyrano(4, 3,2-cdlindazole-2- ethanamine; N-methyl-5-nitro-2H-pyrido(3',2': 5, 6jthiopyrano(4, 3,2-cd]indazole-2- ethanamine; N-methyl-5-nitro-2H-pyrido[4', 5,6]thiopyrano(4, 3,2-cd]indazole-2- 47 N-methyl-5-nitro-2H-pyrido(2', 5,6Ithiopyr~no(4, 3,2-cdlindazole-2- ethanamine; (11) 5-nitro-2H-pyricio(3', 5,6]thiopyrano(4, 3,2-cdlindazole-2-ethanamine; (12) 5-nitro-2H-pyrido[3',2':5,6]thiopyrano(4, 3,2-cdjindazole-2-ethanamine; (13) 5-nitro-2H-pyrido(4', 5,6]thiopyrano(4, 3,2-cd]indazole-2-ethanamine; (14) 5-nitro-2H-pyrido(2', 5,6]thiopyranof4, 3,2-cd]indazol e-2-ethanamine; N-hydroxyethyl-5-nitro-2H-pyrido(3' :5,6]thiopyrano(4, 3,2-cd]indazole-2- ethanamine; (16) N-hydroxyethyl-5-nitro-2H-pyrido(3',2':5, 6]thiopyranof4, 3,2-cd]indazole-2- ethanamine; (17) N-hydroxyethyl-5-nitro-2H-pyrido(4', 3':5,6lthiopyrano(4, 3,2-cd]indazole-2- ethanamine; (18) N-hydroxyethyl-5-nitro-2H-pyrido(2', 6]thiopyranof4, 3,2-cd]indazole-2- ethanamine. 15 8. The compound of claim 1 selected from the group consisting of: N, N-diethyl-5-amino-2H-pyrido[3',4':5,6]thiopyrano[4, 3,2-cd]indazole-2- ethanamine, N, N-diethyl-5-amino-2H-pyrido(3',2': 5,6jthiopyrano[4, 3,2-cdlindazole-2- ethanamine; N, N-dimethyl-5-amino-2H-pyridof 5,6]thiopyrano(4, 3,2-cd]indazole-2- ethanamine; 4 N, N-.diethyl-5-amino-2H-pyrido(4', 5,6]thiopyranof4, 3,2-cd]indazole-2- ethanamine; N, N-dimethyt-5-amino-2H-pyridof2', 5, 6jthiopyrano(4, 3,2-cdlindazole-2- ethanamine; N, N-diethyl-5-amino-2H-pyrido(2',3':5,6]thiopyranof4,3,2-cdlindazole-2- ethanamine; N-methyl-5-amino-2H-pyrido[3',4':5,6thiopyrano4,3,2-cd]indazle-2- ethanamine; N-methyl-5-amino-2H-pyridof3' 5, 6lthiopyranof4, 3,2-cd]indazole-2- STN-methyl-5-amino-2H-pyrido[4', 5, 6lthiopyranot4, 3, 2-cd]indazole-2- e namine; 48 N-methyl-5-amino-2H-pyrido(2',3':5,6lthiopyrano4,3,2-cdjindazoe-.2- ethanamine; (11) 5-amino-2H-pyrido(3',4': 5, 6jthiopyrano[4, 3,2-cdlindazole-2-ethanamine; (12) 5-amino-2H-pyrido(3',2': 5, 6]thiopyrano(4, 3,2-cdjindazole-2-ethanamine; (13) 5-amino- 2 H-pyrido 4 3':5,6thiopyranof4,32-cdindazole2--thanamine; (14) 5-amino-2H-pyrido(2', 5, 6lthiopyrano[4, 3 ,2-cclindazole-2-ethanamine; N-hydroxyethyl-5-amino-2H-pyrido[3', 4':5,6Ithiopyranof4,3,2-cdindzole2- ethanamine; (16) N-hydroxyethyl-5-amino-2H-pyrido3,2': 5, 6]thiopyrano[4, 3,2-cd]indazole-2- ethanamine; (17) N-hydroxyethyl-5-amino-2H-pyridof4', 5,6jthiopyrano(4, 3,2-cd jindazole-2- ethanamine; (18) N-hydroxyethyl-5-amino-2H-pyrido(2',3':5,6]thiopyrano[4,3,2-cdjindazole.2- ethanamine. 9. A method of making a compound of formula according to any one of :claimsi1to 8comprising: a) reacting a compound of formula (1ll): N0 0 U 20 wherein X, Y, Z and T are as defined in claim 1 2a U is selected from the group consisting of F or CI, with a hydrazine of formula (IV): H 2 N-NH-8' (IV) wherein B' has the same meanings as B is defined in formula or B' is a group that can be converted into 8 by removal of protective groups for the primary or secondary amines and hydroxy groups optionally present in to give a compound of formula (11): 49 NO 2 Z JT (II) N--N.B, in which X, Y, Z, T and B' are as above defined. The method of claim 9 further comprising removing protecting groups optionally present to give the compounds of formula in which D is a nitro group. 11. The method of claim 9 or claim 10 further comprising a) reducing the nitro group of intermediate (II) to give the intermediate of formula 10 (lla): NHl NI N--NB converting the compounds of formula in which D is an amino group by removal of the protecting groups optionally present. 12. A pharmaceutical composition comprising the compound of any one of claims I to 8 and a pharmaceutically acceptable excipient. 13. A method of treating tumors in a patient in need of such treatment comprising administering an anti-tumor effective amount of the compound of any one of claims 1 to 8 to the patient. 14. Use of the compound of any one of claims 1 to 8 as an anti-tumor compound. Compounds of formula methods of making them, pharmaceutical compositions containing them or methods of treatment or uses involving them H15.RS1 1 October 1999 4 I substantially as hereinbefore described with reference to the non-comparative examples. DATED: 1 October 1999 CARTER SMITH BEADLE Patent Attorneys for the Applicant: UNIVERSITY OF VERMONT F:#27315.RS1 1 October 1999