GB1603228A

GB1603228A – Indole derivatives a process for preparing them and pharmaceutical compositions containing them
– Google Patents

GB1603228A – Indole derivatives a process for preparing them and pharmaceutical compositions containing them
– Google Patents
Indole derivatives a process for preparing them and pharmaceutical compositions containing them

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Publication number
GB1603228A

GB1603228A
GB14337/78A
GB1433778A
GB1603228A
GB 1603228 A
GB1603228 A
GB 1603228A
GB 14337/78 A
GB14337/78 A
GB 14337/78A
GB 1433778 A
GB1433778 A
GB 1433778A
GB 1603228 A
GB1603228 A
GB 1603228A
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United Kingdom
Prior art keywords
formula
compound
group
dipyrido
methyl
Prior art date
1977-04-13
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)

Expired

Application number
GB14337/78A
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Bpifrance Financement SA

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Agence National de Valorisation de la Recherche ANVAR
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1977-04-13
Filing date
1978-04-12
Publication date
1981-11-18

1978-04-12
Application filed by Agence National de Valorisation de la Recherche ANVAR
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Agence National de Valorisation de la Recherche ANVAR

1981-11-18
Publication of GB1603228A
publication
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patent/GB1603228A/en

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legal-status
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C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07D—HETEROCYCLIC COMPOUNDS

C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 – C07D477/00

C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 – C07D477/00 in which the condensed system contains two hetero rings

C07D487/04—Ortho-condensed systems

A—HUMAN NECESSITIES

A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE

A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics

A61P31/04—Antibacterial agents

A—HUMAN NECESSITIES

A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE

A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

A61P35/00—Antineoplastic agents

A—HUMAN NECESSITIES

A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE

A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

A61P35/00—Antineoplastic agents

A61P35/02—Antineoplastic agents specific for leukemia

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS

C07C205/00—Compounds containing nitro groups bound to a carbon skeleton

C07C205/44—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by —CHO groups

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS

C07C205/00—Compounds containing nitro groups bound to a carbon skeleton

C07C205/49—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups

C07C205/56—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups having nitro groups bound to carbon atoms of six-membered aromatic rings and carboxyl groups bound to acyclic carbon atoms of the carbon skeleton

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS

C07C247/00—Compounds containing azido groups

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07D—HETEROCYCLIC COMPOUNDS

C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems

C07D217/22—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the nitrogen-containing ring

C07D217/24—Oxygen atoms

C—CHEMISTRY; METALLURGY

C07—ORGANIC CHEMISTRY

C07D—HETEROCYCLIC COMPOUNDS

C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 – C07D477/00

C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 – C07D477/00 in which the condensed system contains three hetero rings

C07D487/14—Ortho-condensed systems

Description

PATENT SPECIFICATION ( 1) 1603228
00 ( 21) Application No 14337/78 ( 22) Filed 12 April 1978 > ( 31) Convention Application No7 711 148 ( ( 32) Filed 13 April 1977 in ( 33) France (FR) ( 44) Complete Specification published 18 Nov 1981 ( 51) INT CL 3 C 07 D 471/14; A 61 K 31/47 (C 07 D 471/14, 209/00, 221/00) ( 52) Index at acceptance C 2 C 136 X 145 X 1530 1535 200 213 214 215 220 227 22 Y 246 247 250 251 252 25 Y 281 28 X 290 29 X 29 Y 305 Y 313 31 Y 321 322 323 32 Y 332 337 339 342 34 Y 350 351 352 366 367 386 453 456 45 Y 591 601 611 617 618 620 621 625 628 62 X 630 650 658 660 661 670 671 680 681 761 762 776 802 80 Y AA KK KS KV LK LP LT MK NF TR TT ZF ( 72) Inventors EMILE BISAGNI, CLAIRE DUCROCQ,
CHRISTIAN RIVALLE, PIERRE TAMBOURIN, FRANCOISE WENDLING, JEAN-CLAUDE CHERMANN and LUC MONTAGNIER ( 54) INDOLE DERIVATIVES, A PROCESS FOR PREPARING THEM, AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM ( 71) We, AGENCE NATIONALE DE VALORISATION DE LA RECHERCHE (A N V A R) a body corporate existing under the laws of France of 13, rue Madeleine Michelis, 92522 Neuilly Sur Seine, France, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the 5 following statement:-
The present invention relates to dipyrido l 4,3-bl l 3,4-fl indoles and a process for obtaining thereof The invention also relates to pharmaceutical compositions containing the dipyrido l 4,3-bl l 3,4-fl indoles and to certain therapeutic applications thereof 10 It is known that 9-methoxy-ellipticine, its O-demethylated derivative and, especially, 2-N-methyl-9-hydroxy-ellipticinium acetate, have therapeutic properties which are interesting in the field of cancerology; these compounds are pyrido l 4,3-bl carbazoles of general formula Ia and Ib CH 3 CH 3) C 9 5 OOC RO/ / \ / N-cH 15 NO > NOA j H CH 3 R CH 3, R (la) (Ib) in which R represents CH 3 or H.
To this end, reference may be made to the following articles:
-J B LE PECQ, C GOSSE, NGUYEN DAT XUONG, S CROS and C.
PAOLETTI: Cancer Research 36 p 3067-3076 ( 1976) and -J B LE PECQ, C.
GOSSE, NGUYEN DAT XUONG, and C PAOLETTI: C R Acad Sci Paris 20 56 rie D 281 p 1365 ( 1975).
2 1,603,228 2 Novel compounds, dipyrido l 4,3-b l 3,4-fl indoles, have now been found, which also have therapeutic properties which are interesting in the field of cancerology.
The dipyrido l 4,3-bl l 3,4-fl indoles according to the present invention correspond to formula II:
R RI H CH 13 in which: R’, is hydrogen, hydroxy group, an alkyl group, preferably a lower alkyl group, an alkylthio or alkoxy group, a halogen, such as chloro, or an amino group; R’2 is hydrogen or a lower alkyl group, preferably a methyl group.
In the present specification, the term “lower alkyl” denotes those alkyl groups having from I to 3 carbon atoms, and preferably the methyl group 10 By way of suitable amino groups, mention may be made of those of the following formula:
/ O NH-CH-(CH 2) N R’ 6 R’5 in which N is between 1 and 3, R’6 is hydrogen or a lower alkyl group, for example CH 3 group and R’4 and R’5 are identical or different and each represent an atom of 15 hydrogen or a lower alkyl group; groups R’4 and R’s are preferably identical and are hydrogen, the methyl group or the ethyl group.
The invention also relates to the pharmaceutically acceptable salts of the dipyrido-indoles of formula (II) as well as their isomeric and tautomeric forms when they exist 20 The present invention also relates to a process for obtaining said dipyridol 4,3bl l 3,4-fl indoles.
The process according to the present invention comprises the following steps of:
1) reacting a 6-amino-isoquinoline with 3-nitro-4-chloro-pyridine to form the 25 corresponding 6-l 4 ‘-( 3 ‘-nitropyridyl)aminolisoquinoline; 2) hydrogenating said isoquinoline thus obtained into the corresponding amino compound; 3) reacting the corresponding amino compound with sodium nitrite to form the corresponding triazolopyridine; 30 4) converting the triazolopyridine thus obtained into the corresponding dipyrido l 4,3-bl l 3,4-fl indole, and 5) optionally forming a pharmaceutically acceptable salt of the dipyridoindole thus obtained.
The process of the invention schema:
R 12 R 1 CI N Hp C 13 (II I) (Iv) 1,603,228 may be represented by the following reaction RIC RW 1 W CM 3 (V) Step a Stap 3 F (VII) i tep 4 (VI) Step 5 CH 3 (VIII) z-alky L grup (Tix) The starting materials used in the process of the invention are therefore the 3nitro-4-chloro-pyridine of formula (III) and the 6-amino-isoquinolines of formula (IV).
Step 1: of the process of the invention consists in condensing the 3nitro-4chloro-pyridine of formula III with a 6-amino-isoquinoline of formula IV This condensation may be effected according to different processes well known to the man skilled in the art, these processes varying according to the nature of the substituents R’, and R’2 of the 6-amino-isoquinoline used in this step.
Thus, when the starting 6-amino-isoquinoline comprises no hydroxy groups, step I may be effected in the following manner:
The 6-amino-isoquinoline is dissolved in a suitable inert solvent, such as 1,2dimethoxy-ethane, a solution of dry acid, such as hydrochloric acid, in a dry solvent, is added, then the 3-nitro-4-chloro-pyridine is added The thus obtained reaction mixture is maintained at reflux until one of the reagents has substantially completely disappeared, this being determined by thin layer chromatography measurement; the solvent is then evaporated.
When the starting 6-amino isoquinoline comprises a hydroxy group, it is advantageous to operate at ambient temperature; the starting materials may be dissolved in an inert solvent, such as for example dimethylformamide The reaction mixture obtained by mixture of the solutions of the two starting materials are left at ambient temperature until disappearance of the starting materials which are visible in thin layer chromatography on silica gel The precipitate formed is then recovered by conventional techniques.
Equimolar or substantially equimolar quantities of the two starting materials are advantageously used.
Step 2 of the process according to the invention consists in a hydrogenation of the 6-l 4 ‘-( 3 ‘-nitropyridyl)aminolisoquinoline obtained according to step I described hereinabove This hydrogenation is effected in the presence of a hydrogenation catalyst, such as palladium charcoal The 6-l 4 ‘-( 3 ‘nitropyridyl)aminolisoquinoline is dissolved in an organic solvent, such as for example acetic acid; a suitable quantity of palladium charcoal is added to this solution and the mixture is stirred in a hydrogen atmosphere until the theoretical 5 absorption of hydrogen The catalyst is then eliminated by filtration; the solvent is evaporated and the residue obtained is recrystallised from an organic solvent, such as methanol, ethanol, acetonitrile, xylene, etc.
According to step 3) of the process of the invention, the corresponding triazolopyridine is obtained by treating the 6-l 4 ‘-( 3 ‘-amino-pyridyl)aminol 10 isoquinoline of formula VI with sodium nitrite It is advantageous to carry out this step as follows: the 6-l 4 ‘-( 3 ‘-amino-pyridyl)aminolisoquinoline is dissolved in an organic acid, such as acetic acid, the mixture obtained is cooled to about O C and an aqueous solution of sodium nitrite in the minimum of water is progressively added The reaction mixture is stirred until the temperature returns is to ambient; the precipitate formed is then washed and dried according to conventional methods.
According to step 4) of the process of the invention, the triazolopyridine obtained according to step 3) is converted into the corresponding dipyrido l 4,3bl l 3,4-fl indole In the course of this conversion, an opening of the triazolo ring 20 and a cyclisation are produced to form a dipyrido-indole of formula VIII This step may be carried out in an inert agent, such as paraffin or phenanthrene, having a fairly high boiling point to allow the conversion indicated to occur thermally.
This step is usually carried out at a temperature of 320-350 C.
Examples for carrying out this step will be given hereinafter 25 The dipyrido l 4,3-bl l 3,4-fl indole thus obtained is then possibly converted into a pharmaceutically acceptable salt.
To form these pharmaceutically acceptable salts, it is important to use the suitable agents well known to the man skilled in the art, such as hydrochloric, hydrobromic, succinic, lactic, acetic, phosphoric acids and all other acids 30 commonly used for forming such salts.
The reaction schemes of the preferred proceedings for carrying out the process according to the invention will be indicated hereafter:
Proceedings n l:
This proceeding relates to the obtaining of dipyrido-indoles of formula II 35 hereinabove in which R’, is hydrogen and R’2 is an alkyl group, for example the methyl group; this proceeding is represented by the following diagram for the compound of formula II in which R’2 is a methyl group:
cg.3 N H 2 N ccl CH 5 I 4 CH 3 H CH 3 ( 3) Ci) ( 5) N N / t f NN CH 5 C( 3 ( O CH 3 N N < 4 C 3 ( 8) H C 13 1,603,228 This proceeding will now be illustrated by Examples 1 to 4 hereinafter. Proceeding N 2: This proceeding relates to the obtaining of dipyrido-indoles of formula II hereinabove in which R'1 is the hydroxy group and R'2 is hydrogen or a lower alkyl group. \ '/\ CON 3 CO CH 3 CH 3 N 1 // co 3) 013) R'2 o H CH 3 ( 15) RI? O h NH ( 17) NH co c H 53 R? O 'c H 3. NH O( 4) Nit CO CA C-143 ( 14) cf/Ncl R'a OH N C 3 HA c H 3 ( 33 RI o H II N CH 3 ( 18) Rc R H CH 3 (x$X) The compounds 19 to 26 described hereinafter in the illustrative examples have been synthesised according to this proceeding, in compounds 19 to 25, R'2 = H and R is as defined hereinbelow: 1,603,228 1,603,228 compound 19 R = OH compound 20 R = Cl /compound 21 R =NH-CHCHCHcompound 21 R =NH CH 2 CH 2 CH 2 N C 2 H C 2 H 5 CH 3 / compound 22 R =NH-CH 2 CH 2 CH 2-N CH 3 / compound 23 R =NH-CH 2 CH 2-N CH 3 compound 24 R = NH -CH (CH 2)3-N(C 2 H 5)2 CH 3 compound 25 R = NH-CH 2 -CH 2-CH 2-NH 2; in compound 26 R'2 = CH 3 and R = OH According to this proceeding, the product of formula 15, which is a starting material according to the process of the invention and which may be in its tautomeric form ( 15 a) Ra OH 1 ^ ( 15) R'a O Ha H C/S CH 3 a) may be prepared from cinnamic acid of formula ( 12), by the process which comprises the steps of forming the corresponding azide ( 13); of effecting a cyclisation to form the corresponding isoquinolone and finally of eliminating the protector group of the amino group to obtain the 5-methyl-6-aminoisoquinolone of formula ( 15) The proceeding 2 is then carried out according to the process defined previously to obtain the compound according to the invention of formula XIX in which R is the hydroxy group (compound 19) The hydroxy group of this compound may then be substituted by an atom of chlorine, an amino group, such as for example the following groups, to obtain compounds 19 to 26. C 2 H 5 -NH-(CH 2)3-N C 2 H 5 C 2 H 5 / NH-CH (CH 2)3-N C 2 H 5 CH 3 / -NH (CH 2)n N CH 3 n = 2 or 3 CH 3 -NH-CH 2-CH 2-CH 2-NH 2 CH 3 7 0,2 The cinnamic acid of formula ( 12) used as starting product in proceeding n 2 defined hereinabove may be obtained in different ways Three proceedings will be indicated hereinafter which are suitable for obtaining the cinnamic acid of formula ( 12) lproceedings (a) to (c)l Rg R$ 2 1 5 PN CN Nii CN N CMC CIA CO C 93 cc) S C c C 3 CH 3 ( 11) ( O oa) C 10 b) C CH Co c H 3 R'2 being as defined previously C 3 1) According to this proceeding, the cinnamic acid of formula ( 12) is obtained from the 2-methyl-3-amino-benzonitrile of formula ( 10 a) by the process which comprises the steps of converting the amino group of this compound into the acetylamino group to form the compound of formula ( 10 b); of converting the 10 cyano group of the 2-methyl-3-acetylamino-benzonitrile into the aldehyde group to form the compound of formula ( 11) and then of condensing the aldehyde group of this latter compound with malonic acid to form the cinnamic acid of formula ( 12). This proceeding is illustrated by example 5 hereinafter. O O NHNO CH COOH 15 CH 3 CM 3 ( 30) ( 3 i) N Ha / oo H C" 3 C ( 33) RI? R '2 NHA I COOH co ca c 3/ oc CO) CH 3) CJ 4 & ( 12) ( 32) R'2 being as defined previously. 1,603,228 This proceeding comprises the steps of converting 2-methyl-3-nitroaniline of formula ( 30) into ( 2-methyl-3-nitro-phenyl)chloropropionic acid of formula ( 31), of converting this latter compound into 2-methyl-3-nitro-cinnamic acid of formula ( 32) by elimination of HCI and then in converting the nitro group of this compound into the acetylamino group, passing through the stage of the amino group (formula 5 33) to form the compound of formula ( 12). This proceeding will be illustrated by example 13 hereinafter. Proceeding (c) This proceeding is suitable for obtaining the compound of formula ( 12) in which R'2 is -CH 3 10 C 3 CH 3 CU 3 NO C H 2 Cl No Co NO>c H COOH CHS CIS CH 3 (A) (B) (C) CH 3 CH 3 NH I \coo% CIA COOK c.3 co H 3 CH 3 ( 12) (t) This proceeding comprises the steps of converting the chloro-methyl group of the compound of formula (A) into an aldehyde group lcompound of formula (B)l, of condensing this aldehyde group with malonic acid to form the cinnamic acid of formula (C), of reducing the nitro group of the compound of formula (C) into an 15 amino group lcompound of formula (D)l which is then converted into the acetylamino group to obtain the compound of formula ( 12) This proceeding will be illustrated by example 12 hereinafter.
This proceeding comprises the steps of converting the chloro-methyl group of the compound of formula (A) into an aldehyde group lcompound of formula (B)l, 20 of condensing this aldehyde group with malonic acid to form the cinnamic acid of formula (C), of reducing the nitro group of the compound of formula (C) into an amino group lcompound of formula (D)l which is then converted into the acetylamino group to obtain the compound of formula ( 12) This proceeding will be illustrated by example 12 hereinafter 25 Among the compounds of the present invention, particular mention may be made of the following:
5,1 1-dimethyl-dipyrido l 4,3-bl l 3,4-fl indole; 2,5,9,11-tetramethyl dipyrido l 4,3-bl l 3,4-f 1 indolinium diacetate; 5,1 1-dimethyl-dipyrido l 4,3-bl l 3,4-fi indole acetate 30 5,1 1-dimethyl-dipyrido l 4,3-bl l 3,4-fl indole dihydrochloride.
1,2-dihydro-l-oxo-5-methyl dipyrido l 4,3-bl l 3,4-fl indole; 1-chloro, 5-methyl dipyrido l 4,3-bl l 3,4-f 1 indole; 1-(a-diethylaminopropyl)-amino-5-methyl-dipyrido l 4,3-bl l 3,4-fl indole; 1-(a-dimethylaminopropyl)amino-5-methyl-dipyrido l 4,3-bl l 3,4-f 1 indole; 35 I-(/3-dimethylaminoethyl) amino-5-methyl-dipyrido l 4,3-bl l 3,4-fl indole 1-la-methyl(S-diethylamino butyl)aminol-5-methyl-,dipyrido l 4,3-bl l 3,4f 1 indole; I-l 8-amino-propyl)aminol-5-methyl-dipyrido l 4,3-bl l 3,4-fl indole; 1,2-dihydro-1 oxo-5,11 dimethyl-dipyrido l 4,3-bl l 3,4-f 1 indole.
The invention also relates to antiviral and antitumoral pharmaceutical 40 compositions containing a therapeutically effective quantity of a compound according to the invention of formula II, in combination with a pharmaceutically inert excipient The pharmaceutical compositions according to the invention may 1,603,228 be particularly in the form of solutions injectable by the intravenous (i v) or intramuscular (i m) route.
The antitumoral properties of the compounds according to the invention have been determined by their curative action on inoculated test L-1210 leukemia This leukemia has in fact enabled numerous active compounds used in human 5 therapeutics to be selected.
lZUBROD C G, Proc Nat Acad Sci 1972, 69, 1042-1047 and SCHEPARTZ SA SCREENING, 1971, CANCER Chemother Rep Part 3 vol 2 p 3 l.
This leukemia is maintained in ascitic form by passage (i p route) on CBD 1 10 (C 57 B 16 x DBA/2)F 1 mice The preparations to be tested are injected by the intraperitoneal route at least one day after the inoculation of the cells (one injection only) The results are expressed in increased life span per cent (ILS %) according to KESSEL et al Cancer Res 1971, 31, 1883-1887) or in percentage of the number of cells killed by the product (the survival of the animals is proportional 15 to the number of cells injected).
The protective power of the compounds according to the invention have also been sought on the Friend viro-induced leukemia (J Exp Med 1957-105 307-318) and it has been determined that the compounds according to the invention are antiviral and antitumoral agents 20 The study of the development of the MOLONEY viro-induced sarcoma (Nat.
Cancer Inst Monograph 22, 139-142, 1966) has also made it possible to determine that the compounds according to the invention are antiviral and antitumoral agents.
The toxic dose and the acute toxicity in vivo, i e the lethal doses LD 100 and 25 LD 50, have also been determined.
Furthermore, it has been shown that the compounds according to the invention are cytotoxic at concentrations included between 0 2 and 10 tm and that compound 21 according to the invention is just as active, or more so, than products presently used as antitumoral agents 30 The invention will now be illustrated by the following non-limiting examples.
Example 1.
Preparation of 5,11-dimethyl dipyrido l 4,3-bl l 3,4-fi indole (compound of formula 8) A 5,8-dimethyl 6-l 4 ‘( 3 ‘-nitro-pyridyl)aminolisoquinoline of formula ( 5) 35 36 g of 5,8-dimethyl-6-amino-isoquinoline of formula ( 4) were dissolved in 1 5 1 of 1,2-dimethoxy-ethane; 33 14 g of 3-nitro-4-chloro-pyridine of formula ( 3) were added, then 95 85 ml of a solution of dry hydrochloric acid in ether assayed at 4 362 M l 2 molar equivalents with respect to the amino-isoquinoline of formula ( 3)l were added 40 After 192 hours of heating with reflux, the solvent was evaporated, the residue was taken up with 1 5 1 of water with stirring for 1 hour and the insoluble precipitate was filtered off to give 1 7 g of 3-nitro-4-hydroxy-pyridine.
With constant checking with a p H metre, potassium carbonate was progressively added to the aqueous phase and the precipitate which appears from 45 p H 4 was filtered off when p H 5 5 was reached After drying, said precipitate was recrystallised from benzene and 17 6 g or 28 6 %, of yellow microcrystals were thus obtained, melting point 206 C, corresponding to the expected compound.
Analysis: C 16 H 14 N 403 C H N 50 % calculated 65 29 4 80 19 04 % found 65 09 4 81 18 86 By making alkaline the mother liquors up to p H 10, 13 g or 36 %, of the aminoisoquinoline of formula ( 4), were recovered, melting point: 149 C after crystallisation 55 B/ 5,8-dimethyl-6-l 4 ‘-( 3 ‘-amino-pyridyl)aminolisoquinoline of formula ( 6).
g of nitrated derivative of formula ( 5) were dissolved in I litre of absolute ethanol, to which were added 2 g of 10 % palladium charcoal and the whole was stirred in a hydrogen atmosphere at ambient pressure and temperature until the theoretical quantity of hydrogen has been absorbed After filtration of the catalyst, 60 1,603,228 the solvent was evaporated and the residue recrystallised from ethanol to give 16 9 g ( 93 7 %) of beige crystals, melting point = 235-250 C with decomposition, corresponding to the expected compound crystallised with 1/2 mole of ethanol.
Analysis: C 16 H,6 N 4, 1/2 C 2 Hs O H C H N 5 % calculated 71 05 6 66 19 50 % found 70 78 6 77 19 18 C/l I-l 6 ‘-( 5 ‘,8 ‘-dimethyl-isoquinolyl)ll 4,5-cltriazolo pyridine of formula ( 7).
16.8 g of the amine of formula ( 6) were dissolved in 300 ml of acetic acid, cooled to about O C and 4 83 g of sodium nitrite dissolved in 150 ml of water were 10 progressively added thereto, drop by drop and in maintaining the cold The reaction mixture was maintained cold, under stirring, for 2 hours, then 1 hour allowing it to return to ambient temperature; the solvent was evaporated, the residue was taken up with 300 ml of water and the insoluble precipitate was filtered.
After recrystallisation from ethanol, 14 8 g ( 84 5 %) of pale yellow crystals were is formed, melting point 215-220 C, corresponding to the expected product.
Analysis: C 16 H 1 a N 5 H N C H N % calculated 69 80 4 76 25 44 % found 69 67 4 77 25 23 20 D/ 5,11-dimethyl-dipyrido l 4,3-bl l 3,4-fl indole of formula ( 8) 12 g of the triazolopyridine of formula ( 7) were mixed with 39 g of paraffin with a melting point of 54-56 C and the whole was heated under a nitrogen atmosphere until the gaseous emission was complete i e for 20 to 25 minutes.
After having left the reaction mixture to cool, 100 ml of heavy petroleum ether 25 (boiling point under normal pressure: 100-140 C), were added, the mixture was heated up to boiling and the insoluble black solid was filtered off This latter was taken up with ethanol in the presence of animal charcoal, filtered, concentrated and filtered in the cold, then recrystallised from pyridine to give 5 4 g ( 40 6 %) of yellow micro-crystals, non-melting at 350 C 30 Analysis: C 16 H 3 N 3 H N C H N % calculated 77 71 5 30 16 99 % found 77 52 5 32 16 98 Example 2 35
Preparation of 2,5,9,11-tetramethyl-dipyridol 4,3-bl l 3,4-fi indolinium of formula IX lX = CH 3 COOl.
247 g of the compound of formula ( 8), obtained according to example 1, were suspended in 750 ml of acetone and heated with reflux for 6 hours, in the presence of a large excess of methyl iodide ( 1,42 g) After addition of the same quantity of 40 methyl iodide, heating was again effected with reflux for 14 hours; the reaction mixture was then cooled and 487 mg ( 92 %) of the insoluble product corresponding to the diiodide lcompound of formula (IX) in which X = Il were filtered off.
Analysis: C 18 H 1,2 N 3 H N 45 % calculated 40 68 3 58 7 91 % found 40 55 3 67 7 88 450 mg of the compound obtained, previously dissolved in 100 ml of water, were passed over an exchange resin column (“DOWEX IX 2 “) containing acetate ions and the solvent was evaporated (The word “Dowex” is a registered Trade 50 Mark) The residue was taken up with isobutyl alcohol to give the expected product in the form of yellow-orange microcrystals, melting point 230-235 C It gave one stain by chromatography on a thin alumina layer with the methanol-water mixture ( 4/1 v/v) as eluent; the nuclear magnetic resonance (NMR) spectrum indicates that it is the expected compound, but its centesimal analysis corresponds to the desired 55 product partially hydrated.
I O 1.603228 Example 3.
Preparation of 5,11-dimethyl dipyrido-l 4,3-bl l 3,4-fi indole 1 g of the compound of formula ( 8), obtained according to example I, and 20 ml of acetic acid were heated at boiling point and the excess of acetic acid was immediately evaporated The residue, taken up with acetone and filtered, yielded 5 Ig of insoluble precipitate in the form of yellow ochre micro-crystals, non-melting at 310 C, and corresponding to the monohydrate of the expected compound.
Analysis: C 18 HN 303 H N C H N % calculated 66 44 5 89 12 92 10 % found 66 58 5 83 12 79 Example 4.
5,11 -dipyrido-l 4,3-bl l 3,4-fi-indole dihydrochloride mg of the base of formula ( 8) obtained according to example 1 were s 15 dissolved in 20 ml of absolute ethanol, 1 ml of ethanol saturated by hydrochloric is acid was added and the solvent was evaporated in a water bath under reduced pressure The residue was taken up with acetone and filtered off to yield 200 mg of ochre micro-crystals, non-melting at 310 C and corresponding to the expected dihydrochloride in the form of monohydrate.
Analysis: C 16 H 17 C 12 N 3 OC H Cl 20 C H Cl N % calculated 56 80 5 02 21 01 12 43 % found 56 34 4 78 21 20 12 56 Example 5.
Preparation of 1,2-dihydro-l-oxo 5-methyl-dipyrido-l 4,3-bl l 3,4-fl indole (com 25 pound 19).
A) 2-methyl-3-acetylamino-benzaldehyde offormula ( 11) 39 g ( 0 3 mole) of 2-methyl-3-amino-benzonitrile were dissolved in 75 ml of acetic acid, 30 ml ( 0 3 mole) of acetic anhydride were added, the mixture was heated with reflux for 5 minutes, and cooled The solid obtained was filtered off 30 and the evaporation of the solvent furnished an additional quantity of solid which was added to the preceding one After recrystallisation of the whole from toluene, 44 g ( 95 %) of colourless flakes were obtained with a melting point of 160 C.
Analysis: C 1 o H 10 N 20 H N 35 C H N 35 % calculated 68 95 5 79 16 08 % found 68 82 5 83 15 99 B/ 2-methyl-3-acetylamino-cinnamic acid of formula ( 12) In a 6 1 three-necked flask were introduced 60 g ( 0 34 mole) of the nitrile of formula ( 10) and 1 litre of 50 % diluted formic acid, then the mixture was heated at 40 boiling point Maintaining the whole with reflux, 120 g of Raney alloy were added at 5 times, at intervals of 30 minutes; the mixture was left 30 minutes more at reflux and the salts and the excess of insoluble reagent were filtered.
The precipitate was washed in hot water and the whole of the filtrate was extracted with chloroform, at least ten times with 500 ml of chloroform each time 45 The evaporation of the organic phase gave a residue which distilled, giving 39 g of a product (boiling point 11 = 210-225 C) corresponding to the mixture of the starting nitrile and the aldehyde of formula ( 11), i e the 2-methyl-3acetylaminobenzaldehyde If crystallisation was effected from benzene or toluene, fine, colourless needles were obtained, melting point: 124-128 C 50 Analysis: C 10 H 11 NO 2 C H N % calculated 67 78 6 26 7 91 ‘, found 68 07 6 27 8 05 The preceding mixture ( 38 g), crude from distillation, was dissolved in 50 ml of 55 dry pyridine; it was added all at once to a solution of 300 ml of pyridine containing 22.5 g of malonic acid and I ml of piperidine, and the whole was heated with reflux for 1 1/2 hours The pyridine was evaporated under reduced pressure and the residue was treated by a solution of sodium hydroxide in excess in the presence of chloroform 60 1,603,228 12 1,603,228 12 After decantation, the evaporation of the chloroform gave a residue constituted by a fraction of starting nitrile of formula ( 10).
The acidification of the alkaline layer by hydrochloric acid gave the acrylic acid of formula ( 12), which was recrystallised from acetic acid, giving 25 g ( 34 % with respect to the quantity of nitrile of formula ( 10) used of colourless flakes, 5 melting point = 265-267 C.
Analysis: C 12 H,3 NO 2 C H N % calculated 65 74 5 98 6 39 % found 65 58 6 12 6 52 10 C) 2-methyl-3-acetylamino-cinnamoylazide of formula ( 13) The acrylic acid of formula ( 12) ( 36 g) was added to a solution of 17 g of triethylamine in 150 ml of acetone and the whole was cooled up to O C.
Maintaining the temperature below O C, a solution of 24 3 g of ethylchloroformate in 150 ml of acetone was added drop by drop, the mixture was stirred at O C for 1 15 hour, then the solution formed from 16 g of sodium azide and 40 ml of water was progressively added The reaction mixture was again stirred in the cold for 1 hour after the end of the addition and 27 8 g ( 71 %) of a colourless solid corresponding to the desired azide which was decomposed by melting from 150 C, were filtered off.
The evaporation of the acetone of the mother liquors in a water bath under 20 reduced pressure, and not exceeding 30 C gave an additional quantity of the expected compound which was then coloured yellow This product was used in the synthesis without any other purification.
D) 1-hydroxy-5-methyl-6-acetylamino-isoquinoline or 5-methyl-6acetylamino-1-isoquinoline of formula ( 14) 25 The mixture constituted by 1 51 of diphenyl ether and 33 g of tributylamine was heated to 240 C in a 4 1, three-necked flask 41 g of the azide of formula ( 13) previously dried in a vacuum desiccator were placed in suspension in 300 ml of diphenyl ether and the mixture was added to the preceding solution maintained under violent stirring, in small portions but as rapidly as possible and continuing to 30 heat to avoid the temperature dropping to below 220 C. After the end of the addition, heating was again effected to 240 C and
the reaction mixture was maintained at this temperature for 10 minutes, then it was left to cool The precipitate formed was filtered, washed in benzene and recrystallised from ethanol, in which it was very sparingly soluble, to give 25 g ( 69 %) of colourless 35 microcrystals, non-melting at 320 C.
Analysis: C 12 H 2 N 202 H N C H N % calculated 66 65 5 59 12 96 % found 66 98 5 64 12 87 40 E) 5-methyl-6-amino-1-isoquinolone of formula ( 15) The mixture formed by 500 ml of ethanol, 100 ml of hydrochloric acid and 25 g of the compound of formula ( 14) was heated with reflux for 2 1/2 hours The evaporation of the solvent gave a residue which was taken up with hot water, filtered, and the alkalinisation of the filtrate by N sodium hydroxide gave 16 3 g 45 ( 81 %) of a compound which recrystallised in the form of colourless needles, melting point = 260-285 with decomposition.
Analysis: C 10 o H 1 o N 20 C H N % calculated 68 95 5 79 16 08 50 % found 68 97 5 83 15 85 F) 5-methyl-6-l 4 ‘-( 3 ‘-nitro-pyridyl)aminolll-isoquinolone of formula ( 16) 17.4 g of the compound of formula ( 15) were dissolved in 400 ml of dimethyl formamide (DMF); a solution of 15 9 g of chloronitro-pyridine of formula ( 3) in 100 ml of DMF was added and this mixture was left at ambient temperature for 12 days 55 The precipitate formed was filtered off; the evaporation of the solvent under reduced pressure gave an additional quantity of solid and the whole of said latter was taken up with hot water then rendered alkaline by N sodium hydroxide The 13 1,603,228 1 precipitate formed was recrystallised from DMF, giving 21 3 g ( 72 %) of yellow prisms non melting at 330 C and corresponding to the compound of formula 16.
Analysis: C,5 HHN 403 C H N % calculated 60 80 4 03 18 91 5 % found 60 46 4 08 18 62 When the reaction has been effected in the presence of an excess of 1 to 3 % of the 3-nitro-4-chloropyridine of formula ( 3), apart from the product described hereinabove, from 10 to 15 % of a secondary product, less soluble in the DMF, was isolated, which was recrystallised from this solvent in the form of red prisms, non 10 fusible at 330 C It corresponds to 3-nitro-4-amino di lN-1, N-4 ( 1 ‘hydroxy-5 ‘methyl-6 ‘-isoquinolyl) pyridine).
Analysis: C 26 H,9 N 504, 1/2 H 20 C H N % calculated 64 95 4 35 14 38 15 % found 64 72 4 19 14 28 G/ 5-methyl-6-l 4 ‘-( 3 ‘-amino-pyridyl)aminolisoquinolone of formula ( 17) 12.6 g of the preceding nitrated derivative were dissolved in 500 ml of acetic acid; 0 6 g of 10 % palladium charcoal was added and stirring was effected under a hydrogen atmosphere until the theoretical quantity of hydrogen was absorbed 500 20 ml of acetic acid were added, the mixture was heated to dissolve the precipitate formed, the catalyst was filtered, the solvent was evaporated and the residue was dissolved in water After alkalinisation up to p H 9, the precipitate was filtered and recrystallized in acetonitrile to give 10 2 g ( 84 3 %) of cream microcrystals, corresponding to the hydrate of the amine of formula ( 17) 25 Analysis: C,15 H 16 N 402 C H N % calculated 63 86 5 67 19 71 % found 63 84 5 44 19 47 H/ 1-l 6 ‘-( 1 ‘,2 ‘-dihydro-1 ‘-oxo-5 ‘-methyl-isoquinolyl)ltriazolol 4, 5-cl-pyridine of for 30 mula ( 18).
In a 500 ml, three-necked flask provided with a thermometre, a mechanical stirrer and a dropping funnel, 10 2 g of the amine of formula ( 17) and 70 ml of acetic acid were mixed; the reaction mixture was cooled up to about O C and a solution of 3 g of sodium nitrite in the minimum of water was progressively added Stirring was 35 continued for 1 hour, allowing the reaction mixture to return to ambient temperature; the precipitate was then filtered, washed in water and dried 8 3 g ( 83 5 %)) of colourless microcrystals were thus obtained; melting point 309-310 C, corresponding to the hydrate of the triazolopyridine of formula ( 18).
Analysis: C,5 H,5 N 502 40 C H N % calculated 61 01 4 44 23 72 % found 60 92 4 15 23 44 II 1,2-dihydro-1-oxo-5-methyl-dipyrido l 4,3-bl l 3,4-/7 indole of formula XIX with R = OH: compound 19 45 8 g of triazolopyridine of formula ( 18) were added to 60 g of melted phenanthrene, then heated to 340 C, and the reaction mixture was maintained under stirring at this temperature for 20 minutes, then left to cool The phenanthrene was extracted with petroleum ether or hexane and the insoluble residue was recrystallised from DMF to give 4 2 g ( 58 %) of grey microcrystals non 50 fusible at 310 C and corresponding to the hemi-hydrate of the product of formula ( 19).
Analysis: C,5 H,N 30, 1 l/2 H 20 C H N % calculated 69 75 4 68 16 27 55 % found 70 04 4 40 16 14 1,603 228 Example 6.
1-chloro-5-methyl-dipyrido l 4,3-bl l 3,4-fi indole (compound of formula XIX with R = Cl: compound 20) 1.5 g of the dipyrido-indole (compound 19) was mixed with 250 ml of phosphorus oxychloride containing 1 5 g of phosphorus pentachloride and the mixture was heated with reflux for 20 hours The excess of oxychloride and pentachloride was eliminated in a water bath under reduced pressure and the residue was taken up with tepid water, on several times and in stirring on each time for 1 hour, until exhaustion The aqueous filtrates combined and cooled were neutralised by a solution of sodium or potassium carbonate and the precipitate 10 formed, filtered off then dried, was recrystallised from DMF, giving 875 mg ( 54 %) of yellow micro-crystals, non-melting at 320 C and corresponding to the hemihydrate of the chlorinated derivative 20.
Analysis: C 15 H 10 N 3 C 1, 1/2 H 20 C H N Cl 15 % calculated 65 10 3 97 15 19 12 84 % found 64 77 3 92 14 97 13 21 Example 7.
1-( 8-diethylaminopropyl)amino-5-methyl-dipyrido-l 4,3-bl l 3,4-fl indole (compound of formula XIX with 20 C 2 H 5 / R=-NH-CH 2-CH 2-CH 2-N i e compound 21 C 2 H 5 The mixture, constituted by 875 mg of the chlorinated derivative obtained according to example 6 hereinabove and 10 g of 8-diethyl-aminopropylamine, was heated in an oil bath at 150 C for 30 minutes and the excess of amine was eliminated in a water bath under reduced pressure The residue was extracted three 25 times with 60 ml of boiling benzene and the insoluble residue was taken up with chloroform in the presence of sodium hydroxide After washing of the chloroform layer in water, the chloroform was evaporated, the residue taken up with the benzene previously used and the whole was concentrated up to about 50 ml and cooled The filtration of the solid in suspension gave 180 mg ( 15 %) of yellow 30 microcrystals, melting point = 215-218 C, corresponding to the expected amine crystallised with a molecule of water.
Analysis: C 22 H 27 N,H 20C H N C H N % calculated 69 63 7 70 18 46 35 %found 70 02 7 39 18 29 It has been ascertained that by operating with an excess of 8diethylaminopropylamine, the yield may be improved According to this embodiment, the mixture, constituted by 4 g of the chlorinated derivative obtained according to example 6 hereinabove and 100 ml of y-diethylaminopropylamine, was heated 40 with reflux for 4 hours and the excess of amine was eliminated under reduced pressure The residue obtained was taken up with a N sodium hydroxide solution and the precipitate formed, filtered off and dried, was recrystallised from xylene to give 4 1 g ( 73 %) of the expected derivative which has crystallised with a molecule of water 45 Analysis: C 22 H 27 N 5 H 20 C H N % calculated 69 63 7 70 18 45 % found 69 82 7 49 18 33 1,603,228 Example 8.
1-( 8-dimethylaminopropyl)amino-5-methyl-dipyrido-l 4,3-bl l 3,4-f indole (compound of formula XIX) 3 CH R=-NH-CH 2-CH 2-CH 2-N (compound 22) CH 3 The procedure according to the first proceeding described in example 7 was carried out, in heating at boiling point dimethylamino-propylamine for 7 hours 5 After a treatment identical to the one mentioned hereinabove, the product was recrystallised from benzene to give pale yellow micro-crystals, melting point:
240 C, corresponding to the hemi-hydrate of the expected product.
Analysis: C 20 H 23 N 5, 1/2 H 20 C H N 10 % calculated 70 09 7 00 20 44 % found 70 27 6 85 20 13 By operating according to the embodiment described in example 7 hereinabove, a yield of 73 % has been obtained According to this embodiment, the S 15 chlorinated derivative of example 6 with a large excess of 8dimethylaminopropylamine was heated with reflux for 4 hours After elimination of the excess of amine under reduced pressure, the product was recrystallised from benzene and 73 % of micro-crystals were obtained, the melting point of which is 240 C; this product corresponds to the hemi-hydroxide of the expected product 20 1 g of the compound obtained hereinabove was dissolved in 30 ml of ethanol saturated by hydrochloric acid, the mixture was heated to boiling point and cooled immediately The precipitate formed is recrystallised from ethanol and 1 g of colourless needles was obtained, melting point = 266-268 C, corresponding to the hydrated tri-hydrochloride of compound 22 hereinabove 25 Analysis: C 20 H 23 N 5, 3 HC 1, H 20 C H N Cl % calculated 52 11 6 08 15 20 23 12 % found 51 72 6 21 14 83 22 88 Example 9.
I -(/p-dimethylaminoethyl)amino-5-methyl-dipyrido-l 4,3-bl l 3,4-f indole (compound 30 of formula XIX) CH 3 R=-NH-CH 2 CH 2-N (compound 23) CH 3 The procedure was the same as in examples 7 and 8 above, in heating the chlorinated compound 20, in p-dimethylaminoethylamine at boiling point for 15 35 hours After a treatment identical to those mentioned in the two preceding cases, the product was recrystallised, then taken up with hydrochloric ethanol to form the corresponding trihydrochloride, which was recrystallised from ethanol in colourless prisms, melting point 262-269 C, corresponding to the dihydroxide of the trihydrochloride of the expected compound 23: C,9 H 21 N 5 Yield 37 % 40 Analysis: C,9 H 2,N 5, 3 HCL 2 H 20 C H N Cl % calculated 49 08 6 02 15 07 22 93 % found 49 58 5 77 14 52 23 26 Example 10 45
I-la-methyl-a -diethylamino-butyl)-aminol-5-methyl-dipyrido l 4,3-bl l 3, 4-fl indole (compound of formula XIX with R = NH-CH-CH 2-CH 2-CH 2-N(C 2 H 5)2 i.e compound 24).
500 mg of the chlorinated derivative obtained according to example 6 hereinabove were placed in 10 ml of 2-amino-5-diethylamino-pentane and the 50 1,603,228 16 1,603228 16 mixture was heated with reflux of the amine for 13 hours, under a nitrogen atmosphere and protected from the light The excess of amine was eliminated and the residue was taken up with a N sodium hydroxide solution The precipitate formed was filtered off, washed in water, dried and recrystallised from toluene to give 120 mg ( 17 %) of yellow micro-crystals, melting at around 160 C 5 Analysis: C 24 H 31 N 5 H 20 = 405 5 C H N % calculated 71 08 7 71 17 22 % found 70 78 7 94 17 59 Example 11 10
1-l( 3-amino-propyl)aminol-5-methyl dipyrido l 4,3-bl l 3,4-fl indole.
Compound of formula XIX with R= NH-CH 2-CH 2-CH 2-NH 2 i e.
compound 25).
The procedure is the same as in example 7 with 400 mg of the chlorinated derivative obtained according to example 6 and 10 ml of 1,3-diaminopropane, in 15 heating with reflux for 1 hour The excess of 1,3-diamino-propane was eliminated, the residue was taken up with a N-sodium hydroxide solution and the precipitate was filtered off This latter was recrystallised from dimethylformamide to give 190 mg ( 42 %) of pale yellow micro-crystals, melting point = 268-269 C.
Analysis: C 18 H 1 g Ns, 0 33 H 20 = 311 20 C H N % calculated 69 45 6 32 22 51 % found 69 33 6 50 22 43 Example 12.
Preparation of 1,2-dihydro-l-oxo-5,11-dimethyl-dipyrido-l 4,3-bl l 3,4-fl indole 25 (compound of formula L: compound 26).
The general schema of the synthesis of this compound is shown hereinafter.
This synthesis was carried out according to the proceeding (c) to obtain the compound of formula E(reactions 1 to 4) which is a particular compound of general formula ( 12) and according to proceeding n 2 (reactions 5 to 11) Compounds (G) 30 to (L) hereinafter may be presented in tautomeric form.
CH 3 CH 3 CU 43 e C o.
oee zc% No? HO NO \coo.
CA 3 C 3 CH 3 (A) () (Cc) CH 3 CH CH 3 CIA CH CH NH H C Oo NCH coo H N Hy C CON 3 H 3 c C’3 CH 3 CE) (F)C CD Cs: () (F) 1,603228 1.603 228 CH 3 O CH 3 o 6 o c Nz, 7 M II 1 7, 1 CO C 3 (H 3 f(&) (H) CH 3 o NN CH 3 (X) CH 3 O ff NW CH 3 (I) CCH 3 O N NH (K) c 043 CH 3 O IN NU 11 t H NN CH 3 (L) CH 3 (h) N O C Ha Ct C.H 3 (A) No? ( 5) 2,5-dimethyl-3-nitro-benzaldehyde (B) The mixture constituted by 2,5-dimethyl-3-nitro-benzyl-chloride (A) prepared according to M J WINCHESTER and F D POPP, J Het Chem 12, p 547 ( 1975) ( 610 g), acetic acid ( 1280 ml, water ( 1280 ml) and hexamethylene tetramine ( 855 g) was heated with reflux under stirring for 2 hours 1037 ml of concentrated hydrochloric acid were then added in 10 minutes and the mixture was again heated with reflux for 20 minutes This mixture, cooled to 0 C, gave a solid which was filtered off, dried and recrystallised from cyclohexane ( 4 litres) to give 290 5 g ( 53 0,) of yellow needles corresponding to the aldehyde of formula B, melting point:
= 90 93 C Analysis: Cg Hg NO 3 = 179 o’ calculated o/ found CH 3 Nom C 1 C 60.30 60.13 a H 5.06 4.97 N 7.80 7.71 CH 3 N oe Co CH 3 H (C) (C) 117 1 1 2,5-dimethyl-3-nitro-trans cinnamic acid (C) The mixture constituted by 193 7 g of aldehyde of formula B, 112 5 g of malonic acid, 1 5 1 of pyridine dried on potassium hydroxide and 9 ml of piperidine was heated with reflux for 24 hours but adding, twice, 112 5 of malonic acid, after 3 1/2 hours and 6 hours of reflux After evaporation of the solvent, the residue was 5 taken up with acetone, filtered off, washed in water and again in acetone to give the pure compound C, which was recrystallised from ethanol and gave ochre microcrystals, melting point: 228 C Yield: 173 g ( 72 %).
Analysis: C 1,H,1 NO 4 = 221 21 C H N 10 % calculated: 59 72 5 01 6 33 % found 59 74 4 91 6 21 c HI 3 /” 3 / 2 J Coo H Coo IH CH 3 H c H 3 (C’) (D) CH 3 N t 4 H/6 \COOH Co CH 3 O C 14 C 3 E) 2,5-dimethyl-3-acetylamino-trans cinnamic acid (E) 141 g of nitrocinnamic acid of formula C were suspended in 1260 ml of acetic 15 acid; 170 g of Raney nickel washed in acetic acid and hydrogen were added to this mixture, the mixture being stirred in a hydrogen atmosphere under normal pressure until cessation of absorption of said hydrogen (The theoretical quantity was absorbed) Hot filtration was effected to eliminate the catalyst and half of the acetic acid was evaporated 20 A small sample was evaporated to dryness, taken up with water, neutralised in ammonia, filtered off and recrystallised from ethanol to obtained colourless microcrystals corresponding to 2,5-dimethyl-3-amino-trans cinnamic acid of formula D, melting point 185 C.
Analysis: C 1,H 3 NO 2, 1/2 H 20 = 200 23 25 C H N % calculated: 65 98 7 05 7 00 % found 65 62 6 82 7 07 1,603,228 To the rest, coming from the evaporation of half of the acetic acid, 150 ml of acetic anhydride were added, the resulting mixture was heated with reflux for 1 1/2 hours then evaporated to dryness The solid residue was taken up with hydrochloric water, stirred for 1 hour and filtered off to give a solid which was recrystallised from acetic acid, giving 126 2 g ( 84 %) of colourless flakes corresponding to the acid 5 of formula E, melting point 270 C.
Analysis: C,3 H,5 NO 3 = 233 C H N % calculated 66 93 6 48 6 01 % found 66 78 6 51 6 11 10 CHI’ CH 3 CH 3 HH 3 N NH coo o,, co c HHOW I coc CO CO CH (E) (F) 2,5-dimethyl-3-acetylamino-trans cinnamoylazide (F) The mixture formed by 125 g of cinnamic acid E, 1 1 litre of acetone and 54 g of triethylamine was cooled up to O C, then 78 8 g of ethylchloroformate dissolved in 460 ml of acetone were progressively added with stirring and maintaining the 15 temperature at O C Stirring was continued for one hour at O and in continuing to cool to maintain the temperature below 5 C, a solution of 52 5 g of sodium azide in the minimum of water was added After the end of the addition, stirring was again effected for 1 hour at 0 C, the mixture was allowed to return to ambient temperature, then poured in 5 1 of water and the precipitate formed was filtered off 20 This latter was washed abundantly in water, in terminating the washing with distilled water, then with a little acetone and finally dried to obtain 107 g ( 77 %) of fine cottony needles which melt from 150 C and present one stain only in thin layer chromatography on silica gel This compound F is used as such in the following synthesis 25 CH 3 CH 3 O HyCOs NHXNH I N IHCO 3 C Co CH 3 CH 3 CI 3 H CM 3 C 2 1,603,228 (Q) F) 1,603,228 20 1,2-dihydro-l-oxo-5,8-dimethyl-6-acetylamino-isoquinoline (G) To the mixture constituted by 500 ml of diphenylether and 28 6 g of tributylamine heated at 240 C and maintained under violent stirring, were progressively added, in 15 minutes, 39 6 g of azide in suspension in 450 ml of diphenylether maintained at 40 C, care being taken that the temperature does not drop below 235 C After the end of the addition, stirring was effected for a further minutes at 240 C then the mixture is allowed to cool, eliminating part of the diphenyl ether under vacuum The crystallisation of a solid was observed, 350 ml of benzene were added and the precipitate was filtered off Said latter was taken up with 400 ml of boiling ethanol and the insoluble fraction, filtered off, was recrystallised from 400 ml of dimethylformamide, unfiltering hot, to obtain 18,7 g ( 53 %) of colourless flakes corresponding to compound G.
Analysis: C 13 H 14 N 202: 230 3 % calculated % found CM 3 O NHH CO CH 3 CM 3 (G) C 67.80 67.54 H N 6.13 12 17 6.42 11 96 CH 3 O c NH -7, CH 3 (H) 1,2-dihydro 1 l-oxo-5,8-dimethyl-6-amino-isoquinoline (H) The mixture consisting of 10 6 g of the compound G, 175 ml of ethanol and 35 ml of concentrated hydrochloric acid was heated with reflux for 2 1/2 hours, 300 ml of water were added, the mixture was again heated to boiling point and filtered to eliminate a small insoluble fraction To the cooled filtrate was added a N sodium hydroxide solution to bring the p H to 9 and the precipitate formed was filtered off then recrystallised from ethanol to give 7 35 g ( 85 %) of cream flakes corresponding to the amine of formula H, melting point 242 C.
Analysis: C 11 H 12 N 20 = 188 2 C H N % calculated 70 18 6 43 14 88 %found 70 25 6 15 14 52 CH 3 0 CH 3 t 4 H NH 2 c$ 3 () CH 3 O C NU CM” 3 (I 1) 1,603,228 1,2-dihydro- 11-oxo-5,8-dimethyl-6-l 4 ‘-( 3 ‘-nitro-pyridyl) aminolisoquinoline or 5,8dimethyl-6-l 4 ‘-( 3 ‘-nitropyridyl)aminol isoquinoline (I) The mixture consisting of 34 7 g of the amine of formula H, 27 3 g of 3nitro-4chloro-pyridine and 1 litre of dimethylformamide was stirred at ambient temperature for 15 days and the solvent was evaporated The residue was taken up 5 with 3 litres of N hydrochloric acid and the insoluble fraction was filtered off It corresponds essentially to an undesirable secondary compound which recrystallised from dimethylsulfoxide in form of orange-red microcrystals, non melting at 300 C.
Analysis: C 27 H 23 N 504,H 20 = 499 5 10 C H N %o calculated 64 92 5 04 14 02 % found 65 19 4 81 13 79 To the aqueous phase was added a N sodium hydroxide solution to bring the p H to 9-10 and the precipitate formed was filtered off, then recrystallised from 15 dimethylformamide to give 22 g ( 39 %) of yellow micro-crystals corresponding to the compound of formula I, melting point 310-315 C.
Analysis: C 16 H,N 403 = 310 3 C H N % calculated 61 93 4 55 18 06 20 % found 61 53 4 71 17 76 CH 3 O N N % > N H N 10 MHH Cit 3 C Hi 3 ( 3) ( 3 K) 1,2-dihydro- 11-oxo-5,8-dimethyl 6-l 4 ‘-( 3 ‘-amino-pyridyl) aminolisoquinoline J and 1-l 6 ‘-( 14,2 ‘-dihydro-1 ‘-oxo-5 ‘,8 ‘-dimethylisoquinolyl)l triazolo( 4,5 c) pyridine (K) To 16 8 g of nitrated derivative I in I litre of acetic acid were added 17 g of 25 Raney nickel and hydrogen, stirring the whole under a hydrogen atmosphere at ambient temperature and at normal pressure The theoretical quantity of hydrogen was absorbed in about 1 hour and the catalyst was filtered, then 50 ml of the resultant solution were taken 30 By evaporating this solution, a residue was obtained which was taken up with water and rendered alkaline by the addition of a N sodium hydroxide solution The precipitate formed was filtered off, it was recrystallised from acetonitrile, then from anisole to obtain colourless micro-crystals corresponding to the amine of formula J, melting point 212-215 C 35 Analysis: C,6 H,6 N 4 OH 20 = 298 C H N % calculated 64 41 6 08 18 78 % found 64 14 5 83 18 83 1,603,228 The remaining solution was cooled to 14 C and, with stirring, a solution of 3 61 g of sodium nitrite in the minimum of water was added with 15 minutes Stirring was continued for 1 1/2 hours, allowing the mixture to return to ambient temperature, the solvent was eliminated, the residue was taken up with water, filtered off and the precipitate washed in water The solid obtained was taken up with 1 5 1 of boiling ethanol, filtering was effected, then concentration to 600 ml to give, after cooling, 12 g ( 80 %) of cream microcrystals corresponding to the triazolopyridine of formula K, melting point 300-302 C.
Analysis: C,6 H 13 Ns O O = 291 C H N 10 C H N 10 % calculated 65 97 4 5 24 04 % found 65 66 4 70 24 39 C Hb o CH 3 (L) c 3 1,2-dihydro 1 -oxo-5,11-dimethyl-dipyrido-l 4,3-bl l 3,4-fl indole L 16 g of triazolopyridine of formula K were mixed with 80 g of phenanthrene, 15 and in maintaining the whole under stirring, the mixture was heated in a metal bath at 340 C for 30 minutes, after which time the emission of nitrogen substantially stopped Heating was then effected at 360 C for 2 minutes, the mixture was allowed to cool and poured in 600 ml of hexane The insoluble precipitate was filtered off, washed inboiling hexane, then recrystallised from dimethylformamide 20 to give 5 7 g ( 36 5 %) of grey needles, non-melting at 330 C.
Analysis: Cs H,3 N 3 O 1/2 H 20: 272 3 C H N % calculated 70 57 5 18 15 43 % found 70 63 5 32 15 22 25 Example 13.
This example illustrates the proceedings b) for obtaining the compound of formula ( 12) in which R’2 is hydrogen, i e the compound of formula ( 12 a) hereinafter NO? NC 2 N/2 C 2 Coo Ne CO 30 CH 3 CH 3 CH 3 ( 30 oa) ( 31 a) (% 2 a) 2-methyl-3-nitro-trans cinnamic acid ( 32 a) In a 1 litre, 3-necked flask were introduced 15 2 g of 2-methyl-3-nitro aniline ( 30 a) which were dissolved in 200 ml of acetone and 17 ml of concentrated hydrochloric acid were added, then the whole was cooled to O C.
By maintaining the solution below 5 C, and with stirring, diazotisation was 35 effected by progressively adding a solution of 7 5 g of sodium nitrite in 25 ml of water.
After having left the preceding solution with stirring for 30 minutes, it was poured slowly into a 3-necked flask containing a mixture consisting of 100 ml of acrylic acid, 7 6 g of cupric chloride dissolved in 25 ml of water and 100 ml of 40 acetone, the whole being maintained at a temperature of 35 C for the whole duration of the addition which lasts from 15 to 25 minutes.
This new mixture was maintained with stirring at 35 C for one hour then the acetone and the excess of acrylic acid were evaporated The residue was taken up with chloroform, washed in water and the chloroform phase was exhausted with a 45 cold 2 N sodium hydroxide solution (twice 50 ml) By acidification with 1,603,228 23 1,603,228 23 hydrochloric acid, the ( 2-methyl-3-nitro-phenyl)-chloro-propionic acid ( 31 a) was precipitated, filtered off and dried.
The latter was entirely treated by heating it with reflux for 30 minutes in 100 ml of methanol containing 10 g of potassium hydroxide and, after evaporation, the residue was taken up with water and acidified in the cold by hydrochloric acid The 5 precipitate was filtered, dried and recrystallised from xylene to give 2methyl-3nitro-cinnamic acid ( 32 c), melting point 222 C.
Yield (with respect to the amine used):ll 9 g ( 57 5 %).
MO \Co OH COOK CH 3 co CH 3 (c H 3 ( 12 a) 2-methyl-3-acetylamino-trans cinnamic acid ( 12 a) 10 In a 2-litre 3-necked flask, 84 5 g of nitro-acid ( 32 a) were dissolved in 750 ml of acetic acid, 100 g of commercially available Raney nickel washed with acetic acid and hydrogen were added, stirring being effected under a hydrogen atmosphere at normal pressure and at ambient temperature.
Hydrogenation was stopped when the volume of hydrogen consumed 15 exceeded the theoretical volume ( 28 4 1 instead of the necessary 27 4) or after 4 hours 25 minutes stirring.
The catalyst was filtered off, was washed with acetic acid and half of the solvent was evaporated, 90 ml of acetic anhydride were then added and the mixture was heated with reflux for 1/2 hours then evaporated to dryness 20 The residue was taken up with 500 ml of boiling acetic acid, filtering was carried out in the hot to eliminate the insoluble Ni salts and, on cooling, 44 g of the expected acid were obtained.
By concentration of the mother liquors, about 10 g of the desired compound ( 12 a) were further recovered, melting point 265-267 C The total yield was a 25 minimum of 54 g ( 60 2 %).
Pharmacological tests The following tests were made in groups of 10 mice, unless stipulated to the contrary.
Test 1: Study of the antitumoral properties of the compounds of the invention on L 1210 30 leukemia The antitumoral properties of the compounds according to the invention were determined by their curative action on test inoculated L 1210 leukemia.
This leukemia was maintained on CBD 1 (C 57 B 16 x DBA/2) Fl mice The compounds to be tested were injected by the IP route one or more days after the 35 inoculation of the cells (single injection) The results obtained, shown in table I hereinafter, are expressed in percentage of increase of life span (ILS %) and in percentage of the number of cells killed by the compound to be tested, the survival of the animals being proportional to the number of cells injected.
The percentage of the like span ILS % (Cancer Res 1971, 31, 1883-1887) is 40 the following ratio:
St Sc ILS %= = = 100; Sc St = survival of the treated animals Sc = survival of the control animals The results of table I show that the products according to the invention possess antitumoral properties.
Test 2 Viro-induced Leukemia: Friend leukemia The protective power of the compounds according to the invention was sought on Friend leukemia, inoculated into DBA 2 mice aged from 5-6 weeks The viral 5 inoculum was produced from a homogenate of leukemic spleen (p/volume) diluted to 1/250 in isotonic phosphate buffer without Ca++ nor Mg++ (PBS), corresponding to 100 SD 50 (i e to the dose of virus which induces a splenomegaly in 50 %o of the inoculated mice).
The virus was injected by the IP route and in a volume of 0 2 ml The product 10 to be tested was injected 5 hours after the virus or 1 day after the virus at the dose indicated in a volume of 0 1 ml (IP route) Each group of twenty control mice received the virus and a placebo or the product to be tested, 10 mice were sacrificed on the 21st day, and their spleen removed and weighed The mice were considered as leukemic when the weight of their spleen exceed 200 mg For the 10 15 mice, the life span of the animals was determined.
The results obtained as shown in table II These results show that these products possess an antiviral activity in addition to their antitumoral activity.
Test 3: Study of the development of the viro-induced Moloney sarcoma The injection of Moloney virus lC Jasmin et al J Nat Cancer Inst 1974 53 20 469-474 l by the intramuscular i m route into new-born mice led after 10 days to the formation of a sarcoma The appearance of the tumour was proportional to the dose of virus injected The viral inoculum was constituted by a tumour homogenate diluted to 1/250, this corresponding to 10 TID 50, i e to the dose of virus inducing the appearance of a tumour in 50 % of the animals infected At this dose, 80 to 100 % 25 of the animals developed a tumour, and 100 % of the survivors became leukemic.
The experiment thus consisted in noting the number of mice presenting tumours,the regression, or not, of these tumours and finally in sacrificing the surviving animals after 2 months and in noting the presence or absence of splenomegaly (evidence of a leukemia) The virus was inoculated into new-born mice of 3 to 5 30 days by the IM route, and the mice received the following day (J + I 1) or five hours after injection (JO + 5 hours) the product to be tested by the IP route.
The results obtained are as follows:
Controls virus: 80 % of the young mice presented tumours 10 % of the tumours regressed 3 %,, of the animals survived and all presented a leukemia.
Compound of example 7: injection on J + I; 1 /ug/mouse ( 0 5 mg/kg) % of the mice presented tumours % of the tumours regressed 90 % of the animals survived 40 0 % presented a leukemia.
HUM: injection on J + 1; 5 jtg/mouse ( 2 5 mg/kg) % of the animals present a tumour % regression 10 % of the mice were leukemic 45 HUM: injection on J + 1, 1 jug/mouse ( 0 5 mg/kg) % of the animals present a tumour % regression % survivors 33 % were leukemic 50 Controls virus:
% of the animals present a tumour % of the tumours regressed % of the mice survived 45 % of the mice were leukemic 55 Compond of example 7: injection on JO + 5 hours; 1,ug/mouse ( 0.5 mg/kg).
% of the animals present a tumour % regressed 100 % survived 60 % present a leukemia.
1,603,228 1,603,228 25 Compound of example 1: injection on JO + 5 hours; 1 tug/mouse ( 0.5 mg/kg) % of the animals present a tumour % regressed 75 % survived % are leukemic 5 Test 4: Investigation of the toxicity The toxic dose has been sought, on adult (C 57, BL 6 x DBA/2) FL mice and on new-born mice, of the compounds of the invention by injection by the IP route.
The results obtained are given hereinafter: 10 1 On adult Fl mice (C 57 BL 6 x DBA/2)F 1 (IP route) a) Compound of example 1:
Injection of 2 mg/mouse ( 80 mg/kg) = 5 dead /6 I mg/mouse ( 40 mg/kg) = 1 dead/8 0 5 mg/mouse ( 20 mg/kg) = 1 dead/6 15 b) Compound of example 7:
Injection of 1 mg/mouse ( 50 mg/kg) = 1 dead/3 0.6 mg/mouse ( 30 mg/kg) = 5 dead/10 0.5 mg/mouse ( 25 mg/kg) 0 dead/5 0 3 mg/mouse ( 15 mg/kg) = O dead/6 20 The L Dso of the compound of example 7 is therefore 30 mg/kg.
2-On new-born mice (IP route) ug: 2 5 mg/kg = 3 dead/8, mice puny in appearance HUM 10 ug: 5 mg/kg = 3 dead/8, mice puny in appearance.
These results show that the toxic dose of the compound of example 7 is higher 25 than 2 5 mg/kg of mouse.
Test 5 Cytotoxic effect in vitro The cytotoxic effects of the compounds according to the invention were tested in vitro on cultures of hamster, human and mouse cells.
In particular, the BHK 21 strain of hamster cells and a clone derived from this 30 strain transformed by Hamster Sarcoma Virus (H 55 clone) were used.
After detachment by trypsine, the cells were seeded in plastic Petri dishes ( 35 mm diameter) at the concentration of 2 105 cells/dish, in Eagle medium supplemented with “Bactotryptophosphate” Broth, Difco and 10 % calf serum (R.
M STOKER and I MACPHERSON, Virology, 14, 1961, 359) (The words “Eagle” 35 and “Difco” are registered Trade Marks).
hours later, the cells were well attached to the plastic support, and the products to be tested were added, dilutions having been made thereof in water or in DMSO (dimethylsulfoxide) if the product is sparingly soluble in water In this latter case, a control was made with the same final concentration of DMSO in the culture 40 medium.
The state of the cells was examined 24, 48 and 72 hours afterwards.
The results (table III) clearly show that the products are clearly cytotoxic at concentrations from 0 2 to 10 uim, and that the most active product, i e the compound of example 7, is just as active as the already known derivatives, 2 45 methyl-9-hydroxy-ellipticinium acetate and 9-hydroxy-ellipticine.
The effects on the two types of cells, normal and transformed, are similar.
Test 6: Action on the macromolecular syntheses The action of the compounds according to the invention was studied by the incorporation of precursors marked by radioactive isotopes: 50 14 C methyl-thymidine for studying the DNA synthesis 3 H 5 for that of the RNA 3 Hv valine for that of the proteins.
These precursors were given for a period of 30 minutes at varying times after the addition of the product to be tested 55 The incorporation is measured after lysis of the cells for 1 % of sodium dodecylsulfate and precipitation by 5 % of trichloroacetic acid The acidosoluble precipitate was collected on glass fibre Whatman GF/A filters (The word “Whatman” is a registered Trade Mark) The filters were dried and counted in a liquid scintillation spectrometer.
Table IV gives the typical results for the compound of example 1: 5,11dimethyl-dipyrido-l 4,3-bl l 3,4-fllindole.
It is ascertained that this compound reduces very rapidly, in the hours 5 following its addition, the DNA and RNA syntheses and, to a lesser degree, the synthesis of the proteins.
Experiments made on synchronised cells (G TORFIER, J GRUEST & L.
MONTAGNIER Experimental Cell Research 85, 1974, 437) have shown that the compound of example 1 stops, in the first minutes following its addition, the 10 replication of the DNA, both at its initiation and elongation steps.
Test 7: Study of the antitumoral activity in vitro This activity was measured on a tumoral strain derived from the murine leukemia due to the C FRIEND virus.
The tumoral cells multiply in suspension in a “RPMI 1640 ” medium is (Catalogue of GIBCO Bio-cult Ltd, Washington Road, Sandyford Industrial Estate, PAISLEY PA 3 4 EP, Renfrewshire, Scotland) supplemented with 20 % of embryonary calf serum, penicillin and streptomycine The duplication time of these cells is 11 hours The growth fraction is equal to 1 or very close to 1 (all the cells are in the cycle) The cultures are sub-cultured at time t = 0 at the concentration of 2 105 20 cells per ml in Falcon dishes containing 4 ml of medium After twenty four hours, the product to be tested was added, i e at a moment when the cultures are in exponential phase of growth Twenty four hours after the addition of the product, the cells are counted and the percentage of living cells determined by an exclusion test using “trypan” blueTwo doses may therefore be defined 25 1) LD 50 2) LD 100 LD 100 LD 50 Activityx:
LD 50 HUM LD 50 X Reference product:
9 hydroxy-ellipticinium acetomethylate (HUM) 7 10-7 3 10-7 1 compound of example 1 5 10-7 10-7 3 compound of example 2 5 10-6 1 5 10-6 0 2 compound of example 5 6 10-6 210-6 0 15 compound of example 6 5 10-6 10-6 O 3 compound of example 7 5 10-8 10-8 20 x A;; Thi ie-ll f-ti;r, th T Inf T TAT 4 I wth1 cn,t ttl AT” nóf the IIVI ‘,Lty 1111; Air UI 1 k 1 {LIU UI LIIE IV 50 U 1 Vl VA /ILII lkbi WL, U LA V l -50 U product lobe tested) is quite arbitrary and gives an idea of the activities of the different products by comparing them with the HUM, the most active product in the series of the ellipticines.
1,603,228 27 1,603,228 27 Test 8: Acute toxicity in vivo The products to be tested were injected by the IP route into groups of 10 mice at various dilutions Each day, the number of dead animals is noted If the doses are well chosen, a 100 % lethal dose and a 50 % lethal dose have been able to be defined for each product This study is limited to two months Two isogenic mouse strains 5 were systematically used: C 3 H/He and ICFW.
Mouse strain LD 100 LD 50 Compound of example 1 C 3 H 200 mg/kg 100 mg/kg ICFW 200 mg/kg 150 mg/kg Compound of example 7 C 3 H 35 mg/kg 20 mg/kg ICFW 25 mg/kg 15 mg/kg Test 9: Action on chloromonocytary tumoral cells In this test, the action of the products according to the invention on chloromonocytary tumoral cells was determined according to the chloroleukemia of the mouse test (ICFW mouse, co-sanguine CFW strain) isolated in Unit 22 (B.
TAMBOURIN and F WENDLING) of the “Institut National de la Sante et de la Recherche M 6 dicale Paris” (I N S E R M) in a mouse injected with a variant of the Friend virus.
The tumour kills all the animals injected in 19 3 + 2 days This tumour develops in semi-solid ascitic form; the peritoneum is invaded, both by solid tumoral masses and cells in suspension in the ascitic liquid The tumour is transmitted by the cells of the suspension which produce the tumoral masses and the ascitic cells.
Mean survival time Mean survival without treatment time after treatment Compound of formula 8 ( 40 mg/kg 6 hours after inoculation) 18 5 + 2 days 25 3 + 3 1 days Compound 21 ( 1 mg/kg 24 hours after inoculation) 18 9 + 1 8 day 29 0 + 3 0 days Test I O Comparision of the antitumoral activity of the compound according to example 7 with that of known products The compound of example 7, i e 1-(y-diethylaminopropyl)amino-5-methyldipyrido-l 4,3-bl l 3,4-fi indole or known compounds were injected into L 1210 leukemic mice 1 or 3 days after the inoculation of the cells at the doses indicated in tables V to VII and the mean survival time (MST) and the percentage of increased life span (ILS %) were measured.
Indications concerning the known products used in this example and particularly their antitumoral properties, their secondary effects and their use in man may be found in “La Chimiotherapie des Cancers” by G MATHE and M.
KENIS Expansion Scientifique Francaise, 3rd edition, Paris 1975.
Test 11 L 1210 leukemic mice were injected with the compound of example 8 (compound 22), i e 1-(y-dimethylaminopropyl)amino-5-methyl-dipyrido l 4,3bl l 3,4-fl indole, one day after the inoculation of the cells (day J + I) at the doses indicated in table VIII and the mean survival time (MST) and the increase in life span (ILS O) were measured.
This test was carried out with 103, 104 and 105 leukemic cells on day J This test was also carried out with the compound of example 7, i e 1-l(v diethylaminopropyl)aminol-5-methyl-dipyrido l 4,3-bl l 3,4-fi indole (compound 21).
The results obtained are indicated in table VIII This table also shows the number of the surviving mice; a mouse is considered as survivor if it has survived at least 2 to 3 months after the inoculation of the leukemic cells.
Test 12 Study of the protection of the leukemic mice inoculated L 1210 cells) by a single 5 dose or a fractioned dose.
The compound of example 7 i e 1-l(p-diethyl-aminopropyl)aminol 5-methyldipyrido l 4,3-bl l 3,4-fl indole was injected into L 1210 leukemic mice by one injection ( 10 mg/kg at day J + 3) or by fractioned doses ( 2 5 mg/kg each injection on days J + 3, J + 4, J + 5 and J + 6) 10 The results in table IX show that a single dose has greater antitumoral activity than the same dose injected on 4 occasions.
Test 13 Lewis carcinoma For this test, reference may be made to the following works:
“carcinome pulmonaire de Lewis” ( 3 LL), K SUGIURA and C C STOCK” 15 Cancer Res 1955 15 38-51.
La Chimioth 6 rapie des cancers by G MATHE and Y KENIS, Expansion Scientifique’Frangaise, 3rd edition, Paris 1975.
S A SCHEPARTZ Screening, 1971, Cancer Chemiother Rep Part 3 vol 2, page 3 20 In this test, cells ( 106 cells) removed from a tumour-carrying mouse were injected into mice (BDF) by the IM route The mice developed, 10 to 15 days after the inoculation, a tumour at the spot where the injection was made On the other hand, these cells made metastasis in the lung, forming colonies on the pulmonary surface The antitumoral and antimetastasic effect of a product may therefore be 25 judged on mice thus treated.
Investigation of the antimetastasic effect of the compounds of examples 7 and 8 of the invention 108 cells taken from a tumour-carrying mouse were injected by the IM route (left-hand paw) into groups of 10 mice and the compounds to be tested were 30 inoculated by the IP route on day J + 5 by single or fractioned injection The mean survival time (MST) and the increased life span (ILS %) were determined.
Number of injections MST ILS % non-treated controls 25 Mice treated with the 4 injections 27 4 9 6 compound of example 7 J+ 5 + 6 + 10 + 11 2.5 mg/kg each Mice treated with the 1 injection 32 7 30 8 compound of example 7 J+ 5 mg/kg Mice treated with the J+ 5 27 44 9 8 compound of example 8 5 mg/kg It may be concluded that the compounds of examples 7 and 8 give a protective action concerning the metastasic pulmonary invasion of the lung In fact, one treated mouse and one non-treated mouse were sacrificed on day J + 22 and the pulmonary metastases counted: 77 metastases were counted in the nontreated mouse and 35 in the mouse treated on day J + 5 (one injection) with the compound of example 7.
Test 14: viro-induced leukemia: Friend leukemia A As in test 2 hereinabove, the Friend leukemia virus inoculum was obtained from leukemia spleen and diluted to contain 100 S Ds J O 2 ml (SD 50 = spleen enlarging dose 50 per cent) The product to be tested was compound 21 (obtained 1,603,228 according to example 7) It was given by I P route as a single injection 1 day after virus inoculation.
The results obtained are indicated in table Xa Treatment with this compound 21 as well as with compound of example I (see table II) resulted in a significant increase of the life span 5 Half of the mice treated with compound of example 1 according to test 2 or with compound 21 as above explained were sacrificed 21 days after virus inoculation, their spleen removed and weighted The mean spleen weight for the control mice was 2,520 mg ( 1,919-2,866), that of mice treated with 5 mg/kg of compound of example 1 was 2,013 mg ( 887-2,793), with 25 mg/kg 1,803 ( 718-2, 10 635) and with 50 mg/kg 960 ( 373-1,230) For mice inoculated with compound 21, 1 day after the virus, the mean spleen weight was 1,935 mg ( 500-2,950) The decrease of the splenomegaly is in line with the increase of the life span and suggests that the compounds act also as antiviral agents in this system.
TABLE Xa
Action of compound 21 on Friend leukemia Drug (mg/kg) Range of death (in days) MST ILS % Control mice 0 20-56 32 mice treated with compound 21 (ex7) 5 27-52 44 5 39 B The proceeding of test 2 was repeated by using the compound 21 i e ll(ydiethylaminopropyl)aminol-5-methyl-dipyrido l 4,3-bl l 3,4-fi indole obtained according to example 7.
This test was carried out with a single dose of compound 21 ( 20 mg/kg) and 20 with various virus doses.
The results obtained as shown in table Xb.
1,603,228 TABLE Xb
Viro induced leukemia: Friend leukemia Compound 21: single dosage: 20 mg/kg various dosages of virus Survivors 4 Range of months after death(l) MST( 2) ILS%( 3) inoculation Controls virus 1/500 24-38 29 1 0 ( 1000 S Dso) Compound 21 31-51 40 7 40 0 J+ 1 Controls virus 1/2000 25-44 33 8 0 ( 250 S Ds) Compound 21 50-82 55 4 64 2 J+ 1 Controls (virus 1/1000) 18-36 24 3 0 Compound 21 30-54 44 4 82 7 0 J+ 1 ( 1)Range of death:
the first figure indicates the date when the first mice died and the second the date when the latter mice died.
( 2)MST: Mean of survival time.
( 3)ILS % (increased life span) = increase of the life time; it takes care of dead animals MS Tt MS Tc x 100 MS Tc MS Tt = treated animals; MS Tc= control animals Test 15: L 1210 Leukemia The proceeding of test 1 was repeated with the compound of formula 8 obtained according to example 1, i e 5,1 1-dimethyl-dipyrido l 4,3-bl l 3, 4-fl indole.
In this test, the compound of formula 8 was injected four days after the inoculation of the cells ( 50 mg/kg) The results are shown in table XI wherein the results obtained with the same compound injected one day after the cell inoculation are also reported (see experiment n 2 in table I) The results show that the increased life span (ILS %) was higher when the product was given earlier after the cells inoculum.
1,603,228 1,603,228 TABLE XI
Dose Day of mg/kg inoculation 106 O compound of example 1
O compound of example 1
104 O 104 compound of example 1
106 O 106 compound of example 1
O compound of example 1
104 O 104 compound of example 1 + 1 7-12 7 8 9-15 11 5 47 3 8-11 9 3 + 1 10-13 12 85 38 1 0 10-13 11 + 1 11-14 12 3 11 8 + 4 6-7 6 7 8-13 9 34 3 8-12 8 9 + 4 10-11 10 3 0 10-11 10 5 15.7 + 4 10-16 12 14 7 The drug is administered 1 day (+ 1) or 4 days after the cells inoculum.
Compound of example 1 = dimethyl-5,11 dipyrido l 4,3-bl l 3,4-fl indole.
Test 16 Relationship between structure and activity of various dipyrido indoles differing by the R’l substitution.
The R’1 substitution at the 1 position seems to be very important for the protective effect of the derivatives against L 1210 tumor cell growth.
L 1210 leukemia has been maintained by weekly passages on (C 57 B 16 x DBA/2)Fl mice On day 0, mice were inoculated by I P route with various numbers of L 1210 ascitic tumor cells in volumes of 0 1 ml Subsequently as indicated, drugs were administered in Ca++ and Mg++ free PBS solutions (Gibco) or in Mac Coy’s medium (Gibco or Microbiological Associates).
The relationship between the size of leukemic cell inoculum given IP and the host life span was determined in groups of 10 mice after inoculation of 103 to 106 cells in 0 1 ml of Mac Coy’s medium The decrease in the average of host life-span between each consecutive ten-fold increase in size of the leukemic cell inoculum was 2 days According to the method of calculation of Dombernowsky, the doubling time was 0 615 days.
If the R’1 substituting lateral chain is p-diethylaminopropyl (compound 21) or y-dimethylaminopropyl (compound 22), these compounds have similar or higher antitumoral activity As shown in table XII (compound 21) and compound 22) compete favorably with Ellipticinium acetate (HUM) indicating that the pyridoindole structure is more effective than the pyrido-carbazole nucleus (ellipticine).
Cell number Drug Range of death MST (day) ILS % Exp = 2 Exp = 4 TABLE XII
Range Dose Day of of mg/kg inoculation death MST 0 10-14 11 8 ILS days % surv.
O 104 compound 21 104 compound 21 104 HUM 103 O 103 compound 21 103 compound 21 103 HUM 104 O 104 compound 21 104 compound 22 104 compound 22 O compound 22 104 O 104 compound of the example 2 + 1 13-21 16 2 333 0.5 0.5 + 1 11-15 13 54 11 5 + 1 14-20 14 9 22 6 0 13-16 13 8 O + 1 15-22 17 23 2 + 1 14-20 16 2 17 4 + 1 14-20 16 86 22 15 3 0 10-12 10 9 + 1 14-21 18 65.1 + 1 12-15 13 7 25 7 2.5 0 + 1 10-19 14 7 34 9 8-14 9 05 + 1 11-16 13 8 52 5 0 10-13 11 2 + 4 10-17 11 3 0 9 0 Test 17 Relationship between the length of the substituting chain and the activity.
The importance of the substitution of R’l on 1 position for the antitumoral activity of these compounds was demonstrated above If one varies the length of the lateral chain y-diethylaminopropyl-amino 1 for the compound 21 or pdimethylamino-ethylamino 1 for compound 23, as shown in table XIII, both compounds are active against L 1210 leukemia at the same dosage However compound 21 is less toxic ( 50 mg/kg for compound 21 and 25 mg/kg for compound 23); it has a higher ILS% than compound 23 and 1 mouse survived up to 60 days.
These results indicate that the diethylaminopropylamino-1 l substitution corresponds to the best length of chain for the therapeutic action of the derivative.
Increasing the lateral chain by (a-methyl 8-diethylamino-butyl)amino 1 such as compound 24, similar antitumoral effect is found, conforming that compound 21 is the compound the most active on the mice.
Cell number 104 Drug 1,603,228 on.
1,603,228 TABLE XIII
Day of inoculation Range of death MST ILS % Survivors up to 60 days Control 8-9 8 8 Compound 21 + 1 12-18 14 9 69 2 1 Compound 23 + 1 11-18 14 6 65 2 0 Control 10-13 11 2 Compound 21 + 4 14-32 18 5 65 6 0 Compound 24 + 4 12-20 17 5 56 25 0 Test 18 Relationship between dose and L 1210 antitumoral activity of compound 21 The dose response of the activity of compound 21 as shown with L 1210 leukemia according to test 1 is as follows:
At 5 mg/kg, the mean survival time (MST) is significantly increased (ILS = 87 per cent); at higher doses of compound 21, the MST is always significant increased (ILS = 52 p cent at 10 mg and 75 p cent at 20 mg/kg) and some were surviving up to days ( 10 p cent of inoculated mice at 10 mg and 20 p cent at 20 mg per kg).
Test 19 Effect of the time of administration of compound 21 As shown in table XIV, a single injection of compound 21 increased significantly the mean survival time when given as later as 6 days after the L 1210 cells inoculation The results confirm that compound 21 is more effective when given earlier after the cells inoculation (ILS = 56 4 % at day + 2, 43 % at day + 3 instead of 20 8 % at day + 6 or 2 % at day + 7) The non-protective effect at day + 7 could be explained by the fact that some cells migrate through a compartment inaccessible to the drug (as postulated by J B Le Pecq for the 9-hydroxyellipticine).
TABLE XIV
Day of administration Range of death MST ILS % Control 10-11 10 1 BD 40 + 2 15-17 15 8 56 4 105 + 3 12-17 14 4 43 0 5 105 + 4 10-14 12 4 22 8 106 + 5 10-13 11 9 17 8 5 106 + 6 10-14 12 2 20 8 107 + 7 10-13 10 3 2 0 5 107 BD 40 is given I P at 0 1 ml containing 0 4 mg/mice ( 20 mg/kg).
Day 0 is the day of inoculation of 104 L 1210 cells given I P Evaluation of the number of leukemic cells by the mean of calculation of the doubling time (according to P Dombernowsky).
Estimated number of cells Test 20: Study of the combination of compound 21 with other drugs An increase of protection of mice inoculated with Lewis lung carcinoma 14 days before a low priming dose followed by a high challenge dose of endoxan was reported by J Millar 1978, tenth congress of chimiotherapy ( 1978) in press, G A.
Preasant Cancer vol 40 1977 p 987-993 for the therapy of drug-resistant breast 5 carcinoma used a combination of endoxan, BCNU and Adriamycin the following day (The word “Adriamycin” is a registered Trade Mark).
The results concerning similar experiments (breast carcinoma and Lewis carcinoma) with compound 21 are discussed in table XV A priming dose of endoxan ( 100 mg/kg) was given 3 days after 105 leukemic cells ( 20 mice) The 10 following day, 10 mice received 0 1 ml of PBS (control mice) and 10 mice received mg/kg of compound 21 Increase in life span was significantly higher in mice receiving endoxan and compound 21 compared to the mice with endoxan or compound 21 alone The results listed in table XV suggested that compound 21 given one day after endoxan or endoxan-BCNU gave rise to a higher protection of S 15 mice than endoxan, endoxan + BCNU or compound 21 given alone Taking as control mice the endoxan treated mice, the ILS % is 57 for endoxan + compound 21 treated mice and 63 for endoxan + BCNU + compound 21 treated mice.
TABLE XV
Range of death MST ILS % Control 10-13 11 2 Endoxan 11-17 13 1 16 9 BD 40 14-32 18 5 65 2 Endoxan BD 40 18-24 20 6 84 Endoxan + BCNU 10-17 13 7 22 3 Endoxan + BCNU 18-24 21 4 91 + BD 40 BD 40 is given I P at day + 4 ( 20 mg/kg) Endoxan is inoculated I P at day + 3 ( 100 mg/kg) and BCNU immediately after endoxan ( 2 mg/kg).
Day 0 is the day of inoculation of 105 cells given I P.
* Test 21: Toxicity of compounds 21, 22, 23 and 24 20 All drugs were dissolved in aqueous solutions adjusted to p H 5 with acetic acid and inoculated to the mice under a volume of 0 1 ml I P after dilution in Mac Coy’s medium.
The toxicity of the drugs after interperitoneal injection was estimated from the mortality rate up to the 30th day The minimal quantity resulting in 100 % deaths 25 was 50 mg/kg for compound 24,50 mg/kg for compound 21 and for compound 22, mg/kg for compound 23 In general, the deaths occurred early after drug administration ( 4-6 days for compound 24, 4-8 for compound 21, 3-5 for compound 22 and 4-6 for compound 23).
1,603,228 TABLE I
Study of the antitumoral properties of the compounds according to the invention on L 1210 leukemia Number of Quantity in Number of Number of cells mg/mouse cells cells Surviving Compound tested Experiment injected (mg/kg) ILS % found killed mice Compound of experiment 106 1 mg/mouse 34 32 105 90 % 0 example 1 No 1 1 05 ( 50 mg/kg) 15 73 40,000 60 % 0 (formula 8) J+ 2 104 14 78 6,000 40 % 103 3 800 20 % 1/10 experiment 106 1 mg/mouse 47 3 8,000 99 % No 2 105 ( 50 mg/kg) 38 1 2,000 98 % J+ 1 104 11 8 5,000 50 % Compound of experiment 0 1 mg/mouse no significant difference example 1 No 3 0 05 mg/mouse with the controls 104 cells (formula 8) 103 cells compound 20 0 1 mg/mouse 0 1 mg toxic and no difference (example 6) with the controls 104 cells 103 cells ON t’i al TABLE I (Continued) Study of the antitumoral properties of the compounds according to the invention on L 1210 leukemia Number of Quantity in Number of Number of cells mg/mouse ILS cells cells Surviving Compound tested Experiment injected (mg/kg) % found killed mice Compound 21 Product 104 O 1 mg/mouse 33 3 60 99 94 1/10 (example 7) injected by ( 0 5 mg/kg) I.P route J+l 0 1 mg/mouse 11 5 103 90 ( 5 mg/kg) 103 0 01 mg/mouse 17 4 80 92 ( 0.5 mg/kg) 0.1 mg/mouse 23 2 40 96 5/10 ( 5 mg/kg) HUM product 0 1 mg/mouse injected by ( 5 mg/kg) I.P route 104 22 6 400 96 1/10 J+ 1 O 1 mg.
( 5 mg/kg) 103 0 1 mg 22 15 60 94 3/10 ( 5 mg/kg) HUM 2-N-methyl-9-hydroxy-ellipticinium acetate.
J+ 1 or J 42: injection one or two days after inoculation of the cells.
0 0 o 1Oo Viro-induced TABLE II leukemia Friend leukemia Product to be Mean survival time ILS Weight of the spleens Mouse Virus tested Days % on the 21st day in mg Control VFA 1/250 ( 1) 31 7 ( 26-47) 2519 mg VFA 1/250 ( 2) 28 ( 14-17) ( 2866-1919) Treated VFA 1/250 compound of example 1 (formula 8) hrs after the virus 0.1 47 6 ( 26-78) 59 5 2013 mg ( 887-2793) 0.5 34 2 ( 22-54) 14 6 ( 1803 mg) ( 718-2635) 1 mg 41 7 ( 14-81) 39 7 959 6 mg ( 373-1230) Male DBAZ mice aged 5-6 weeks Experiment of 4 8 76 VFA: anaemic Friend virus, injected by the I P route, 0 2 ml dilution 1/250-100 SD 50 Mean survival time between brackets, dates on which the first and last mouse died Increase in the life span per 100 C) Go t’J 38 1,603,228 38 TABLE III
Minimum dose involving complete inhibition of growth Compound Compound Compound Compound Compound example example example example example Cells 1 2 5 6 7 HE C 13/8 0 5 M 2 5 2 0 25 0 25 HS 5 0 5 M 2 5 2 0 25 0 25 HE = 9-OH-hydroxy-ellipticine or 2-acetomethylate-9-hydroxy-ellipticinium.
TABLE IV
Inhibition of the macromolecular syntheses by the compound of example 1
Time, in hours, Incorporation in Incorporation in Incorporation in of marking the DNA (in % of the RNA (in % of proteins (in % of after addition the non-treated the non-treated the non-treated of the product control) control) control) 0.5 41 13 40 3 14 13 37 6 6 3 3 7 16 9 6 9 17 24 4 31 17 the concentration of the product is 5 M.
-the cells H 55 were seeded 24 hours beforehand at a concentration of 5 105 per Petri dish of 60 mm.
1,603,228 TABLE V
Quantity per Compound tested Injection mouse Range of death MTS ILS % Control 104 cells 10-15 days 11 Compound of example 7 J+l 0 2 mg 6 1 mouse alive on 30th day Compound of example 7 J+ 3 0 2 mg 11 17 13 8 25 45 Thiotepa J+ 3 0 1 mg 10 11 11 0 Mitomycine J+ 3 O 016 mg 10 15 11 54 4 95 BCNU J+ 3 0 1 mg 10 12 11 1 0 9 Methotrexate J+ 3 0 250 mg 15 17 15 9 44 62 Endoxan J+ 3 4 mg 13 3 mice alive on 30th day/10 I I ‘0 TABLE VI
Quantity in mg/mouse Injection Range of death MTS ILS % Control 105 cells 9 10 9 06 Thiotepa 0 1 mg J+ 3 9 10 92 1 54 Mitomycine C 0 016 mg J+ 3 9 10 9 06 0 BCNU O 1 mg J+ 3 8 9 8 8 0 Methotrexate 0 250 mg J+ 3 12 14 12 62 39 35 rl J+ 1 9 16 12 6 39 07 Compound of example 7 O 2 mg J+ 3 10 15 12 27 35 43 Endoxan 4 mg J+ 3 15 21 18 6 105 30 TABLE VII
Compound tested Injection Quantity Range of death MTS ILS % Control 105 cells 10 11 Compound of example 7 J+ 3 0 2 mg/mouse 13 16 14 18 36 35 ON COVIN,, 0 5 /tg/mouse 10 13 10 8 3 85 0 C,, ool TABLE VIII
Number of Number of L 1210 cells Compound Quantity injected surviving injected on day J used in mg/mouse MTS ILS % mice 103 Controls 13 Example 8 0 1 mg/mouse 16 4 50 5 3 Example 8 0 05 mg/mouse 14 7 34 9 1 Example 7 0 2 mg/mouse 16 3 25 4 7 Example 7 O 1 mg/mouse 15 15 4 4 104 Controls 10 9 Example 7 0 1 mg/mouse 18 65 1 1Example 8 0 1 mg/mouse 13 7 25 7 3 Example 8 0 05 mg/mouse 14 7 34 9 1 Controls 9 05 Example 8 0 1 mg/mouse 13 8 52 5 0.05 mg/mouse 11 6 28 1 dose in mg/mouse x 50 = dose in mg/kg.
C) ti 0 c N.) TABLE IX
Quantity of the compound Number of L 1210 cells of example 7 used in Surviving injected on day J mg/mouse Injection MTS ILS % mice 104 Controls 11 9 0.2 mg/mouse 4 x ( 0 05 mg/mouse) single at 14 2 19 32 1 J+ 3, fractionated at 14 4 21 O J+ 3 + 4 + 5 + 6 Controls 10 4 0.2 mg/mouse single at 14 18 36 35 4 x ( 0 05 mg/mouse) J+ 3, fractionated at 13 4 28 85 f J+ 3 + 4 + 5 + 6 To a) 0 i hi

Claims (27)

WHAT WE CLAIM IS:-
I Dipyrido l 4,3-bl l 3,4-f) indoles corresponding to the formula Rp 2 Ri o N CH 3 in which:
R’, is hydrogen, the hydroxy group, an alkyl, alkylthio or alkoxy group, a halogen 5 or an amino group; R’2 is hydrogen or a lower alkyl group; and the pharmaceutically acceptable salts thereof.

2 Compound according to claim 1, wherein R’2 is hydrogen.

3 Compound according to claim 1, wherein R’2 is the methyl group 10

4 Compound according to claim 1, wherein R’, is an amino group of formula:
/R’4 / NH-CH-(CH 2)n-N RPO RP 5 wherein N is between 1 and 3, R’6 is hydrogen or a lower alkyl group and R’4 and R’5, identical or different, are hydrogen or a lower alkyl group.

5 Compound according to claim 4, wherein R’, is selected from the group 15 consisting of hydrogen and methyl and R’4 and R’s are identical and selected from the group consisting of hydrogen, methyl and ethyl groups.

6 Dipyrido l 4,3-bl l 3,4-fl indoles corresponding to the formula 49 2 ‘N H 9 8 3 N cx 5 in which: 20 R’, is hydrogen, the hydroxy group, a lower alkyl, a halogen or an amino group of formula:
R’4 NH-CH (CH 2)n-N R’6 R’5 wherein N is between 1 and 3, R’6 is hydrogen or a lower alkyl group and R’4 and R’5, identical or different, are hydrogen or a lower alkyl group; 25 R’2 is hydrogen or a lower alkyl group; and the pharmaceutically acceptable salts thereof.

7 Compound according to claim 4, wherein R’2 is methyl group and R’1 is selected from the group consisting of hydrogen and hydroxy group.

8 Compound according to claim 4, wherein R’2 is hydrogen and R’1 is chloro 30

9 Dipyrido l 4,3-bl l 3,4-fl indoles corresponding the formula:
C 3 Ri 1 N /t ‘N H CH 3 la H C 143 1,603,228 wherein R’, is hydrogen or a hydroxy group.

Dipyrido l 4,3-bl l 3,4-fl indoles corresponding to the formula:
CH 3 RA N, N t< 9 Ia H Cóa wherein R', is a lower alkyl group. 11 Dipyrido l 4,3-bl l 3,4-fl indoles corresponding to the formula CH 3 R 11 Iq N l Ia H CH 53 wherein R'1 is halogen or an amino group of formula: R'4 / NH-CH-(CH 2) -N R'6 R 15 wherein N is between 1 and 3, R'6 is hydrogen or a lower alkyl group and R'4 and R'5, identical or different, are hydrogen or a lower alkyl group 10 12 Dipyrido l 4,3-bl l 3,4-fi indoles corresponding to the formula: R'1 v (MC) Ic H CH 3 wherein R', is hydrogen, hydroxy group, halogen or an amino group of formula R'4 NH-CH-(CH 2),-N I \ R'6 R' 5 wherein N is between I and 3, R'6 is hydrogen or a lower alkyl group and R'4 and 15 R'5, identical or different, are hydrogen or a lower alkyl group. 13 Compound according to claim 6, wherein R'6 is selected from the group consisting of hydrogen and methyl and R'4 and R'5 are identical and selected from the group consisting of hydrogen, methyl and ethyl groups. 14 Compound of claim 1, which is one of the following compounds: 20 5,1 1-dimethyl dipyrido l 4,3-bl l 3,4-fl indole; 2,5,9,1 1-tetramethyl dipyrido l 4,3-bl l 3,4-fi indolinium diacetate; 5,1 1-dimethyl dipyrido l 4,3-bl l 3,4-fl indole acetate; 5,1 1-dimethyl dipyrido l 4,3-bl l 3,4-fl indole dihydrochloride; 1,2-dihydro-l-oxo-5-methyl dipyrido l 4,3-bl l 3,4-fl indole; 25 l-chloro-5-methyl dipyrido l 4,3-bl l 3,4-fl indole; 1 l(y-diethylaminopropyl)aminol 5-methyl dipyrido l 4,3-bl l 3,4-fl indole; Il(y-dimethylaminopropyl)aminol 5-methyl dipyrido l 4,3-bl l 3,4-f 1 indole; 1,603,228 l(ip 3-dimethylaminoethyl)aminol-5-methyl dipyrido l 4,3-bl l 3,4-fl indole; 1 l(a-methyl-a S-diethylamino-butyl)aminol-5-methyl dipyrido l 4,3-bl l 3, 4-fl indole; ll(p-amino-propyl)aminol-5-methyl dipyrido l 4,3-bl l 3,4-f 11 indole; 1,2-dihydro 1-oxo-5, 1 -dimethyl dipyrido l 4,3-bl l 3,4-fi indole 5 Process for obtaining the compounds of Claim 1, comprising the steps of: (I) reacting a 6-amino-isoquinoline with 3-nitro-4-chloro-pyridine to form the corresponding 6-l 4 '-( 3 '-nitropyridyl)aminolisoquinoline; ( 2) hydrogenating said isoquinoline thus obtained into the corresponding amino compound; 10 ( 3) reacting the corresponding amino compound with sodium nitrite to form the corresponding triazolopyridine; ( 4) converting the triazolopyridine thus obtained into the corresponding dipyrido l 4,3-bl l 3,4-fl indole; and ( 5) optionally forming a pharmaceutically acceptable salt of the dipyrido indole 15 thus obtained. 16 Process of claim 15, wherein the hydrogenation of step ( 2) is effected in the presence of a hydrogenation catalyst, such as palladium charcoal. 17 Process of claim 15, wherein step ( 4) is effected in the presence of an inert agent such as paraffin or phenanthrene, of fairly high melting point to allow the 20 reaction to occur thermally. 18 Process of claim 15, wherein the starting product of formula IV in which R'2 is as defined in Claim 1 and R'1 = OH, i e the compound of formula 15 or 15 a (tautomeric form) RIP OH R 12 O 25 0 A 3 5 a ( 15) C 3 is obtained by the process comprising the steps of converting the amino group of 2methyl-3-amino-benzonitrile into the acetylamino group to form the compound of formula 10; of converting the cyano group of 2-methyl 3-acetylaminobenzonitrile into the aldehyde group and then in condensing said latter with malonic acid to form the acid of formula ( 12); of forming the corresponding azide ( 13); in effecting 31 a cyclisation to form the corresponding isoquinolone and of eliminating the protecting group from the amino group to obtain the 5-methyl-6-amino isoquinolone of formula ( 15). 19 The process of Claim 15, wherein the starting product of formula IV in which R', = OH and R'2 is as defined in Claim 1, i e the compound of formula 15 or 3 a (tautomeric form), is obtained by the process comprising the steps of converting the 2-methyl-3-nitro-aniline of formula ( 30) into ( 2-methyl3nitrophenyl)chloropropionic acid of formula ( 31), of converting this latter compound into 2-methyl-3-nitro-cinnamic acid of formula ( 32) by elimination of HCI and then converting the nitro group of this compound into the acetylamino group passing through the stage of the amino group (formula 33) to form the compound of formula ( 12), the compound of formula ( 12) being then converted into the compound of formula 15 or 15 a according to claim 18. The process of Claim 15, wherein the starting product of formula IV in which R', = OH and R'2 is a CH 3 group, i e the compound of formula 15 or 15 a in which R'2 = CH 3, is obtained by the process which comprises the steps of converting the chloromethyl group of the compound of formula (A) into an aldehyde group lcompound of formula (B)l, of condensing this aldehyde group with malonic acid to form the cinnamic acid of formula (C), of reducing the nitro group of the compound of formula (C) into an amino group lcompound of formula (D)l which is then converted into the acetylamino group to obtain the compound of formula ( 12), the compound of formula ( 12) being then converted into the compound of formula ( 15) or ( 15 a) according to claim 18. 21 Antitumoral and antiviral pharmaceutical composition comprising a 1,603,228 therapeutically effective quantity of a compound of Claim 1, in combination with a pharmaceutically inert excipient. 22 Pharmaceutical composition of claim 21, in the form of a solution injectable by the intravenous or intramuscular route. 23 Method of treatment of cancers in non-human animals consisting in 5 administering a therapeutically effective dosage of a compound according to claim 1. 24 Process according to claim 20, wherein the 2,5-dimethyl-3-acetyl-aminotrans cinnamic acid (formula 12) is converted into 1,2-dihydro-l-oxo-5,8dimethyl6-amino-isoquinoline (formula 15 a) by the following steps: 10 1) said acid of formula 12 is converted into the corresponding 2,5dimethyl-3acetylamino-trans cinnamoylazide (formula F) 2) the cyclisation thereof gives the 1,2-dihydro-l-oxo-5,8-dimethyl-6acetylamino-isoquinoline (formula G). 25 Process of claim 15, wherein the 6-amino-isoquinoline used in step I is 1,2 15 dihydro-l-oxo-5,8-dimethyl-6-amino-isoquinoline obtained according to process of claim 26 and the resulting compound of step 1 is 5,8-dimethyl-6-l 4 '(-3 '-nitropyridyl)aminolisoquinoline or 1,2-dihydro 1-oxo-5,8-dimethyl-6-l 4 '-( 3 '-nitropyridyl)aminolisoquinoline; the compound obtained in step 2 is 1,2dihydro-1-oxo5,8-dimethyl-6-l 4 '-( 3 '-aminopyridyl)aminolisoquinoline or its tautomeric form; and 20 the compound obtained in step 3 is 1-l 6 '( 1 ',2 '-dihydro-l'-oxo-5 ',8 '-dimethylisoquinolyl)ltriazolo( 4,5 c)pyridine or its tautomeric form. 26 Process as claimed in claims 15, substantially as described in any one of the foregoing Examples 1 to 12. 27 A dipyrido-indole obtained by a process according to any of claims 15 to 20 25 and 24 to 26. For the Applicants, CARPMAELS & RANSFORD, Chartered Patent Agents, 43 Bloomsbury Square, London WC 1 A 2 RA. Printed for Her Majesty's Stationery Office by the Courier Press, Leamington Spa, 1981. Published by the Patent Office, 25 Southampton Buildings, London, WC 2 A l AY, from which copies may be obtained. 1,603,228 GB14337/78A 1977-04-13 1978-04-12 Indole derivatives a process for preparing them and pharmaceutical compositions containing them Expired GB1603228A (en) Applications Claiming Priority (1) Application Number Priority Date Filing Date Title FR7711148A FR2387229A1 (en) 1977-04-13 1977-04-13 DIPYRIDO (4,3-B) (3,4-F) INDOLES, PROCESS FOR OBTAINING, THERAPEUTIC APPLICATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM Publications (1) Publication Number Publication Date GB1603228A true GB1603228A (en) 1981-11-18 Family ID=9189361 Family Applications (1) Application Number Title Priority Date Filing Date GB14337/78A Expired GB1603228A (en) 1977-04-13 1978-04-12 Indole derivatives a process for preparing them and pharmaceutical compositions containing them Country Status (12) Country Link US (2) US4266060A (en) JP (1) JPS549299A (en) BE (1) BE865851A (en) CA (1) CA1117945A (en) CH (1) CH633796A5 (en) DE (1) DE2815724A1 (en) ES (1) ES468721A1 (en) FR (1) FR2387229A1 (en) GB (1) GB1603228A (en) IT (1) IT1095582B (en) LU (1) LU79422A1 (en) NL (1) NL184895C (en) Families Citing this family (19) * Cited by examiner, † Cited by third party Publication number Priority date Publication date Assignee Title US4334070A (en) * 1980-12-15 1982-06-08 Hoffmann-La Roche Inc. Cycloalka[4,5]pyrrolo[2,3-g]isoquinolines DK158666C (en) * 1982-11-04 1990-11-19 Glaxo Group Ltd ANALOGY PROCEDURE FOR PREPARATION OF 5,14-DIHYDROBENZ-OE5,6AA-ISOINDOLOOE2,1-BAA-ISOQUINOLIN-8,13-DION NO163330C (en) * 1984-05-03 1990-05-09 Glaxo Group Ltd ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY ACTIVE ISOQINOLINE DERIVATIVES. JPH0642591B2 (en) * 1984-11-30 1994-06-01 ミノルタカメラ株式会社 Method for manufacturing printed wiring board NO872963L (en) * 1986-07-29 1988-02-01 Glaxo Group Ltd Isoquinoline derivatives. DK616087A (en) * 1986-11-27 1988-05-28 Glaxo Group Ltd ISOQUINOLINE DERIVATIVES AND PHARMACEUTICAL PREPARATIONS CONTAINING THEM GB8800312D0 (en) * 1988-01-07 1988-02-10 Glaxo Group Ltd Process FR2627493B1 (en) * 1988-02-23 1991-10-31 Sanofi Sa PROCESS FOR THE PREPARATION OF ISOQUINOLEIN DERIVATIVES FR2645861A1 (en) * 1989-04-17 1990-10-19 Inst Nat Sante Rech Med USE OF DIPYRIDO (4,3-B) (3,4-F) INDOLES FOR THE PREPARATION OF MEDICAMENTS USEFUL FOR THE TREATMENT OF AIDS US5281608A (en) * 1992-08-28 1994-01-25 American Home Products Corporation Substituted tetrahydropyrido[3',4':4,5]-pyrrolo[3,2-c]quinolines FR2859474B1 (en) 2003-09-04 2006-01-13 Centre Nat Rech Scient USE OF INDOLE-DERIVED COMPOUNDS FOR THE PREPARATION OF A MEDICAMENT USEFUL IN THE TREATMENT OF GENETIC DISEASES RESULTING FROM THE ALTERATION OF SPLICE PROCESSES JP4932495B2 (en) * 2004-01-23 2012-05-16 アムゲン インコーポレイテッド Compound and method of use EP2266972A1 (en) * 2009-06-12 2010-12-29 Splicos New chemical molecules that inhibit the splicing mechanism for treating diseases resulting from splicing anomalies CN106928205B (en) 2009-06-12 2019-09-24 Abivax公司 For treating the compound of presenility and especially early ageing US10253020B2 (en) 2009-06-12 2019-04-09 Abivax Compounds for preventing, inhibiting, or treating cancer, AIDS and/or premature aging EP2465502A1 (en) 2010-12-15 2012-06-20 Société Splicos Compounds useful for treating AIDS EP2757161A1 (en) 2013-01-17 2014-07-23 Splicos miRNA-124 as a biomarker of viral infection ES2898385T3 (en) 2013-07-05 2022-03-07 Abivax Bicyclic compounds useful for the treatment of diseases caused by retroviruses EP2974729A1 (en) 2014-07-17 2016-01-20 Abivax Quinoline derivatives for use in the treatment of inflammatory diseases Family Cites Families (2) * Cited by examiner, † Cited by third party Publication number Priority date Publication date Assignee Title FR2248828B1 (en) * 1973-10-29 1977-04-15 Anvar US3933827A (en) * 1974-10-29 1976-01-20 Hoffmann-La Roche Inc. 9-Acyloxy-5,11-dimethyl-6H-pyrido[4,3-B]carbazoles 1977 1977-04-13 FR FR7711148A patent/FR2387229A1/en active Granted 1978 1978-04-10 BE BE186694A patent/BE865851A/en not_active IP Right Cessation 1978-04-12 CA CA000301011A patent/CA1117945A/en not_active Expired 1978-04-12 CH CH391878A patent/CH633796A5/en not_active IP Right Cessation 1978-04-12 GB GB14337/78A patent/GB1603228A/en not_active Expired 1978-04-12 LU LU79422A patent/LU79422A1/en unknown 1978-04-12 IT IT22254/78A patent/IT1095582B/en active 1978-04-12 ES ES468721A patent/ES468721A1/en not_active Expired 1978-04-12 DE DE19782815724 patent/DE2815724A1/en active Granted 1978-04-13 NL NLAANVRAGE7803951,A patent/NL184895C/en not_active IP Right Cessation 1978-04-13 US US05/896,101 patent/US4266060A/en not_active Expired - Lifetime 1978-04-13 JP JP4376678A patent/JPS549299A/en active Granted 1980 1980-11-21 US US06/209,091 patent/US4444776A/en not_active Expired - Lifetime Also Published As Publication number Publication date CA1117945A (en) 1982-02-09 IT7822254D0 (en) 1978-04-12 NL7803951A (en) 1978-10-17 LU79422A1 (en) 1979-11-07 JPS549299A (en) 1979-01-24 CH633796A5 (en) 1982-12-31 JPS6225670B2 (en) 1987-06-04 NL184895C (en) 1989-12-01 IT1095582B (en) 1985-08-10 DE2815724C2 (en) 1991-02-14 NL184895B (en) 1989-07-03 FR2387229A1 (en) 1978-11-10 FR2387229B1 (en) 1980-05-09 BE865851A (en) 1978-10-10 US4266060A (en) 1981-05-05 ES468721A1 (en) 1978-12-16 US4444776A (en) 1984-04-24 DE2815724A1 (en) 1978-10-26 Similar Documents Publication Publication Date Title GB1603228A (en) 1981-11-18 Indole derivatives a process for preparing them and pharmaceutical compositions containing them Cholody et al. 1992 Chromophore-modified antineoplastic imidazoacridinones. 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US3303095A (en) 1967-02-07 Compositions and methods for producing tranquilizing activity CN107281180A (en) 2017-10-24 Application of the 8- alkyl berberine salts in prevention and treatment lung-cancer medicament is prepared WO2023208018A1 (en) 2023-11-02 Substituted pyrimidine hydrazide compound, method for preparing same, and use thereof WO1991005770A1 (en) 1991-05-02 Potential anticancer agents derived from acridine Legal Events Date Code Title Description 1982-02-10 PS Patent sealed [section 19, patents act 1949] 1994-12-07 PCNP Patent ceased through non-payment of renewal fee Effective date: 19940412
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