Cytotoxic agents

ES 2 311 639 T3

DESCRIPTION

Cytotoxic agents.

Background of the invention

DNA topoisomerases are enzymes that are present in the nuclei of cells in which they catalyze the breaking and regrouping of DNA strands, which control the topological state of DNA. Recent studies also suggest that topoisomerases are also involved in the regulation of template supercoiling during RNA transcription. There are two main classes of mammalian topoisomerases. DNA topoisomerase I catalyzes changes in the topological state of double-stranded DNA by performing temporary single-strand break-join cycles. In contrast, mammalian topoisomerase II alters DNA topology by causing a transient break of double strands joined by enzyme bridges, followed by passage and ligation of the strands. Mammalian topoisomerase II has been further classified into type II α and type II β. Antitumor activity associated with agents that are topoisomerase poisons is associated with their ability to stabilize the enzyme-DNA cleavable complex. This drug-induced stabilization of the enzyme-DNA cleavable complex effectively converts the enzyme to a cellular venom.

Several antitumor agents in clinical use have potent activity as mammalian topoisomerase II poisons. These include adriamycin, actinomycin D, daunomycin, VP-16, and VM-26 (teniposide or epipodophyllotoxin). Unlike the number of clinical and experimental drugs that act as topoisomerase II poisons, there are currently only a limited number of agents that have been identified as topoisomerase I poisons. Camptothecin and its structurally related analogs are among the most widely studied topoisomerase I poisons. Recently bi- and terbenzimidazoles have been identified (Chen et al., Cancer Res. 1993, 53, 1332-1335; Sun et al., J. Med. Chem. 1995, 38, 3638-3644; Kim et al., J Med. Chem. 1996, 39, 992-998), certain benzo [c] phenanthridine and protoberberine alkaloids and their synthetic analogs (Makhey et al., Med. Chem. Res. 1995, 5, 1-12; Janin and col., J. Med. Chem. 1975,18,708-713; Makhey et al., Bioorg. & Med. Chem. 1996,4,781-791), in addition to the fungal metabolites, bulgarein (Fujii et al., J. Biol. Chem. 1993,268, 13160-13165) and saintopine (Yamashitay col., Biochemistry 1991, 30 , 5838-5845) and indolecarbazoles (Yamashita et al., Biochemistry 1992, 31, 12069-12075) as topoisomerase I poisons. Other topoisomerase poisons have been identified including certain benzo [i] phenanthridine and cinnoline compounds (see LaVoie et al., US Patent No. 6,140,328 (735,037WO1) and WO 01/32631 ( 735.044WO1)). Despite these reports, there is currently a need for additional agents that are useful for treating cancer.

Summary of the invention

Applicant has discovered compounds that show activity against topoisomerase I and / or topoisomerase II and compounds that are effective cytotoxic agents against cancer cells, including drug-resistant cancer cells. Accordingly, the invention provides a compound of the invention which is a compound of formula I:

<img file="ES2311639T3_D0001.tif" />

in which:

A and B are independently N or CH;

Y and Z are independently hydroxy, alkoxy (QC₆), alkoxy (C₁-C₆) substituted, alkanoyloxy (C₁ -C₆), alkanoyloxy (C₁-C₆) substituted, -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; or Y and Z together with the ring carbon atoms to which they are attached form an alkylenedioxy ring with 5 to 7 ring atoms;

Ri is (Ci-C6) alkyl; Y

R_c and R_d are each independently alkyl (C₁-C₆) or alkyl (C₁ -C₆) replaced; or R_c and R_d together with the nitrogen to which they are attached they form an N'-alkyl (C₁ -C₆) piperazino, pyrrolidino or piperidino, ring which may be optionally substituted with one or more aryl, heteroaryl or heterocycle;

or a pharmaceutically acceptable salt thereof.

ES 2 311 639 T3

The invention also provides a compound of the invention which is a compound of formula II:

<img file="ES2311639T3_D0002.tif" />

in which:

A and B are independently N or CH;

W is N or CH;

R₃ and R are both H, or R₃ and R together are = O, = S, = NH or = NR₂ in which R₂ is alkyl (C₁-C₆) or alkyl (C₁C₆) replaced;

Y and Z are independently hydroxy, alkoxy (C₁-C₆), alkoxy (C₁-C₆) substituted, alkanoyloxy (C₁-C₆), alkanoyloxy (C₁-C₆) substituted, -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; or Y and Z together with the ring carbon atoms to which they are attached form an alkylenedioxy ring with 5 to 7 ring atoms;

R₁ is hydrogen or alkyl (C₁-C₆); Y

R_c and R_d are each independently alkyl (C₁ -C₆) or alkyl (C₁ -C₆) replaced; or R_c and R_d together with the nitrogen to which they are attached they form an N'-alkyl (C₁-C₆) piperazino, pyrrolidino or piperidino, ring which may be optionally substituted with one or more aryl, heteroaryl or heterocycle;

provided that at least one or both of A and B is N; and as long as R₃ and R₄ let both be H, then W be CH;

or a pharmaceutically acceptable salt thereof.

The invention also provides a compound of the invention which is a compound of formula III:

<img file="ES2311639T3_D0003.tif" />

in which:

X is O, S, NH or = NR₂ in which R₂ is alkyl (C₁ -C₆) or alkyl (C₁-C₆) replaced;

Y and Z are independently hydroxy, alkoxy (C₁-C₆), alkoxy (C₁-C₆) substituted, alkanoyloxy (C₁-C₆), alkanoyloxy (C₁-C₆) substituted, -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; or Y and Z together with the ring carbon atoms to which they are attached form an alkylenedioxy ring with 5 to 7 ring atoms;

R₁ is hydrogen or alkyl (C₁-C₆); Y

R_c and R_d are each independently alkyl (C₁ -C₆) or alkyl (C₁ -C₆) replaced; or R_c and R_d together with the nitrogen to which they are attached they form an N'-alkyl (C₁-C₆) piperazino, pyrrolidino or piperidino, ring which may be optionally substituted with one or more aryl, heteroaryl or heterocycle;

or a pharmaceutically acceptable salt thereof.

ES 2 311 639 T3

The invention also provides a compound of the invention which is a compound of formula IV:

<img file="ES2311639T3_D0004.tif" />

in which:

A and B are independently N or CH;

W is N or CH;

Y and Z are independently hydroxy, alkoxy (Ci-C₆), alkoxy (C₁-C₆) substituted, alkanoyloxy (C₁ -C₆), alkanoyloxy (C₁-C₆) substituted, -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; or Y and Z together with the ring carbon atoms to which they are attached form an alkylenedioxy ring with 5 to 7 ring atoms;

R_i and R₂ are independently H, alkyl (C₁-C₆) or alkyl (C₁-C₆) replaced; or R_i and R₂ they are together = O or = S; and

R_c and R_d are each independently alkyl (C₁-C₆) or alkyl (C₁ -C₆) replaced; or R_c and R_d together with the nitrogen to which they are attached they form an N'-alkyl (C₁-C₆) piperazino, pyrrolidino or piperidino, which ring may be optionally substituted with one or more aryl, heteroaryl or heterocycle.

The invention also provides a pharmaceutical composition comprising an effective amount of a compound of the invention in combination with a pharmaceutically acceptable diluent or carrier.

The invention also provides a method for modulating topoisomerase activity in a mammal in need of such treatment which comprises administering to the mammal an amount of a compound of the invention effective to provide a topoisomerase modulating effect.

The invention also provides a method of inhibiting the growth of cancer cells which comprises administering to a mammal afflicted with cancer an amount of a compound of the invention effective to inhibit the growth of said cancer cells.

The invention also provides a method which comprises inhibiting the growth of cancer cells by contacting said cancer cell in vitro or in vivo with an amount of a compound of the invention effective to inhibit the growth of said cancer cell.

The invention also provides a compound of the invention for use in medical therapy, preferably for use in the treatment of cancer, for example solid tumors, in addition to the use of a compound of the invention in the preparation of a medicament useful for the treatment of cancer, for example solid tumors.

The invention also provides novel processes and intermediates disclosed herein that are useful in preparing compounds of the invention. Some of the compounds of the invention are useful in preparing other compounds of the invention.

Detailed description

The following definitions are used, unless otherwise described.

"Alkyl (C_i-C₆) "Denotes both straight and branched carbon chains with one or more, for example, 1, 2, 3, 4, 5 or 6, carbon atoms, but reference to a single radical such as" propyl "only encompasses the radical straight chain, specifically referring to a branched chain isomer such as "isopropyl".

"Alkyl (C₁ -C₆) substituted "is an alkyl group of the formula (C₁ -C₆) as defined above wherein one or more (e.g. 1 or 2) carbon atoms in the alkyl chain have been substituted with a heteroatom independently selected from -O-, -S- and NR- (where R is hydrogen or C alkyl₁ -C₆) and / or wherein the alkyl group is substituted with 1 to 5 substituents independently selected from cycloalkyl, substituted cycloalkyl, alkoxy (C₁-C₆) -carbonyl (for example -CO₂Me), cyano, halogen, hydroxy, oxo (= O), carboxy (COOH), aryloxy, heteroaryloxy, heterocyclooxy, nitro, and -NR^toR^b, in which R^to and R^b it can be the same or different and is selected from hydrogen, alkyl, arylalkyl, heteroarylalkyl, heterocycloalkyl, cycloalkyl, substituted cycloalkyl, arite, heteroaryl, and heterocyclic. Alkyl groups (C₁-C₆) substituted are exemplified by, for example, groups such as hydroxymethyl,

ES 2 311 639 T3 hydroxyethyl, hydroxypropyl, 2-aminoethyl, 3-aminopropyl, 2-methylaminoethyl, 3-dimethylaminopropyl, 2-carboxyethyl, hydroxylated alkylamines, such as 2-hydroxyaminoethyl and similar groups. Alkyl groups (C₁-C₆Preferred substituted) are alkyl groups (C₁-C₆) substituted with one or more substituents of formula -NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a nitrogen-containing heterocyclic ring. Specific examples of such heterocyclic rings include piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino. Other alkyl groups (C₁-C₆Preferred substituted) are alkyl groups (C₁-C₆) substituted with one or more heterocyclic rings containing carbon-linked oxygen. Specific examples of such oxygenated heterocyclic rings are, for example, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, and the like groups.

"Alkoxy (C₁-C₆) ”Refers to groups of the formula (C₁-C₆) -O- in which alkyl (C₁-C₆) is as defined in this document. Preferred alkoxy groups include, by way of example, methoxy, ethoxy, propoxy, iso-propoxy, n-butoxy, cer-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like groups.

"Alkoxy (C₁-C₆) substituted "refers to an alkyl group (C₁-C₆) substituted-O- in which alkyl (C₁-C₆) substituted is as defined above. Alkoxy (C₁-C₆) substituted is exemplified by groups such as O-CH₂CH₂-NR_toR_b, O-CH₂CH₂-CHR_toR_b or O-CH₂-CHOH-CH₂-OH and similar groups. Alkoxy groups (C₁-C₆) Preferred substituted are (C₁-C₆) substituted with one or more substituents of formula -NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a heterocyclic ring. Specific examples of such heterocyclic rings include piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino. Other alkoxy groups (C₁-C₆Preferred substituted) are alkoxy groups (C₁-C₆) substituted with one or more heterocyclic rings containing carbon-linked oxygen. Specific examples of preferred oxygenated heterocyclic ring substituents are, for example, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, and like groups. Specific examples of such oxygenated heterocyclic rings are, for example, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, and the like groups.

"Alkanoyloxy (C₁-C₆) ”Includes, by way of example, formyloxy, acetoxy, propanoyloxy, iso-propanoyloxy, n-butanoyloxy, eerc-butanoyloxy, sec-butanoyloxy, n-pentanoyloxy, n-hexanoyloxy, 1,2-dimethylbutanoyloxy, and the like groups.

"Alkanoyloxy (C₁-C₆) substituted "refers to an alkanoyloxy group (C₁-C₆) in which one or more (for example 1 or 2) carbon atoms in the alkyl chain have been substituted with a heteroatom independently selected from -O-, -S- and NR- (in which R is hydrogen or alkyl C₁-C₆) and / or wherein the alkyl group is substituted with 1 to 5 substituents independently selected from cycloalkyl, substituted cycloalkyl, alkoxy (C₁-C₆) -carbonyl (for example -CO₂Me), cyano, halogen, hydroxy, oxo (= O), carboxy (COOH), aryloxy, heteroaryloxy, heterocyclooxy, nitro, and -NR^toR^b, in which R^to and R^b may be the same or different and are selected from hydrogen, alkyl, arylalkyl, heteroarylalkyl, heterocycloalkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclic. Alkanoyloxy (C₁-C₆) substituted is exemplified by groups such as -OC (= O) CH₂-NR_toR_b and OC (= O) -CHOH-CH₂-OH. Alkanoyloxy groups (C₁ -C₆) Preferred substituted are groups in which the alkyl group is substituted with one or more nitrogen and oxygen containing heterocyclic rings such as piperazino, pyrrolidino, piperidino, morpholino, thiomorpholino, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl and the like groups.

Aryl denotes a phenyl radical or an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic. Examples of aryl include phenyl, indenyl, and naphthyl.

Heteroaryl encompasses a radical attached through a ring carbon of a monocyclic aromatic ring containing five or six ring atoms that is made up of carbon and one to four heteroatoms each selected from the group that is made up of non-peroxy oxygen, sulfur, and N ( X), where X is missing or is H, O, alkyl (C₁C₄), phenyl or benzyl, in addition to a radical of an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benz-derivative or one derived from the condensation of a propylene, trimethylene or tetramethylene diradical thereto . Examples of heteroaryl include furyl, imidazolyl, triazolyl, triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl, pyridyl (or its N-oxide), thienyl, pyrimidinyl (or its N-oxide), indolyl isoquinolyl (or its N-oxide) and quinolyl (or its N-oxide).

The term "heterocycle" refers to a monovalent saturated or partially unsaturated cyclic non-aromatic group containing at least one heteroatom, preferably 1 to 4 heteroatoms, selected from nitrogen (NR_x, in which R_x it is hydrogen, alkyl or a direct bond at the point of attachment of the heterocycle group), sulfur, phosphorus and oxygen within at least one cyclic ring and which can be monocyclic or multicyclic. Such heterocycle groups preferably contain 3 to 10 atoms. The point of attachment of the heterocycle group can be a carbon or nitrogen atom. This term also includes heterocycle groups fused to an aryl or heteroaryl group, provided the point of attachment is on a ring containing non-aromatic heteroatoms. Representative heterocycle groups include, by way of example, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, morpholinyl, indolin-3-yl, 2imidazolinyl, 1,2,3,4-tetrahydroisoquinolin-2-yl, uinuclidinyl, and the like.

"Aryloxy" refers to a group of the formula aryl-O-, where aryl is as defined herein. Examples of aryloxy groups include, phenoxy, and 1-naphthyloxy.

"Heteroaryloxy" refers to a group of the formula heteroaryl-O-, where heteroaryl is as defined herein. Examples of heteroaryloxy groups include 3-piperidyloxy, 3-furyloxy, and 4-imidazolidinyl.

ES 2 311 639 T3 "Heterocyclooxy" refers to a group of the formula heterocycle-O-, where heterocycle is as defined herein. Examples of heterocyclooxy groups include 4-morpholinooxy and 3-tetrahydrofuranyloxy.

"Arylalkyl" refers to a group of the formula aryl-alkyl (C₁ -C₆) -, where aryl and alkyl (C₁ -C₆) are as defined in this document.

"Heteroarylalkyl" refers to a group of the formula heteroaryl-alkyl (C₁-C₆) -, wherein heteroaryl and alkyl (C₁C6) are as defined in this document.

"Heterocycloalkyl" refers to a group of the formula heterocyclo-alkyl (C₁ -C₆) -, in which heterocycle and alkyl (C₁-C₆) are as defined in this document.

Specific and preferred values ​​listed below for radicals, substituents, and ranges are for illustration only; they do not exclude other defined values ​​or other values ​​within defined ranges for radicals and substituents.

The following specific values, preferred values, and discussion refer to compounds of formula I.

Specifically, alkyl (C₁ -C₆) can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl or hexyl.

A specific value for A is CH.

Another specific value for A is N.

A specific value for B is N.

Another specific value for B is CH.

A specific value for Y is OH.

Another specific value for Y is alkoxy (C₁-C₆).

Another specific value for Y is -OCH₃.

Another specific value for Y is alkoxy (C₁ -C₆) replaced.

Another specific value for Y is -OCH₂CH₂OH.

Another specific value for Y is -OCH₂CH₂OCH₂CH₃.

Another specific value for Y is -O-CH2-CHOH-CH2-OH.

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Y is -OC (= O) CH2-NRaRb.

Another specific value for Y is -OC (= O) -CHOH-CH2-OH.

Another specific value for Y is alkyl (C₁-C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Y is -OC (= O) CH2-NRaRb.

A specific value for Z is OH.

Another specific value for Z is alkoxy (C₁-C₆).

Another specific value for Z is OCH3.

Another specific value for Z is alkoxy (C₁-C₆) replaced.

Another specific value for Z is -OCH2CH2OH.

ES 2 311 639 T3

Another specific value for Z is -OCH₂CH₂OCH₂CH₃.

Another specific value for Z is -O-CH₂-CHOH-CH₂ -OH.

Another specific value for Z is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Z is -O-CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Z is -OC (= O) -CHOH-CH2-OH.

Another specific value for Z is alkyl (C₁ -C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Z is -OC (= O) CH2-NRaRb.

A specific value for Ri is alkyl (C₁ -C₆).

Another specific value for R1 is methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, n-pentyl, isopentyl, or nhexyl.

A preferred compound of formula (I) is the compound 5-ethyl-8,9-dimethoxy-5H-2,3-dioxa-5,12-diaza-cyclopenta [b] crisen-6-one, 5-butyl-8 , 9-dimethoxy-5H-2,3-dioxa-5,12-diaza-cyclopenta [b] crisen-6-one, or a pharmaceutically acceptable salt thereof.

Certain compounds of formula (I) can act as prodrugs for other compounds of formula (I). For example, a compound of formula (I) in which Y and / or Z is -OP (= O) (OH)₂ or -OC (= O) NR_toR_d; can act as a prodrug for a corresponding compound of formula (I) wherein Y and or Z is hydroxy. Consequently, a specific subset of compounds of formula (I) are compounds in which Y and / or Z is -OP (O) (OH)₂ or -OC (= O) NR_cR_d. A particularly preferred compound is a compound of formula (I) in which Y and / or Z is -OP (= O) (OH)₂. Another preferred compound is a compound of formula (I) in which Y and / or Z is -OC (= O) NR_cR_d, in which R_c and / or R_d is alkyl (C₁-C₆) substituted with one or more -NR_andR_F in which R_and and R_F are each independently alkyl (C₁-C₆). Another preferred compound is a compound of formula (I) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd together with the nitrogen to which they are attached form an N '- (alkyl ) piperazino, pyrrolidino or piperidino. A more preferred compound is a compound of formula (I) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd together with the nitrogen to which they are attached form a piperidino ring, which ring it is optionally substituted with an N-linked heterocycle ring (eg piperidino).

The present invention provides compounds of formula I and a process for preparing compounds of formula I wherein R₁ is alkyl (C₁ -C₆) or alkyl (C₁ -C₆) substituted which comprises reacting the compound of formula I in which R1 is H with a suitable nitrogen alkylating agent, such as an alkyl halide (C1C₆) or alkyl halide (C₁ -C₆) substituted, to form an alkyl compound (C₁ -C₆) or alkyl (C₁ -C₆) substituted corresponding. One skilled in the art understands that the nitrogen atom of lactam can be conveniently manipulated synthetically and efficiently converted to useful related compounds, for example, by preparing intermediates with a protected N atom and whose protected nitrogen atom can be deprotected and subsequently alkylated for providing the aforementioned alkylated nitrogen compounds.

A compound of formula I can be prepared by subjecting a corresponding intermediate of formula A to suitable cyclization conditions, for example, by treatment with palladium acetate and tri-o-tolylphosphine, as illustrated in Scheme 1 below. A compound of formula I can also be prepared by subjecting a corresponding intermediate of formula B to conditions suitable for ring system formation, for example, by treatment with a suitable tin reagent, as illustrated in Scheme 2 below. The compounds of the present invention include intermediates of formulas A and B.

Scheme 1

<img file="ES2311639T3_D0005.tif" />

ES 2 311 639 T3

Scheme 2

<img file="ES2311639T3_D0006.tif" />

Formula B Formula I

Other suitable conditions for the formation of the ring system from intermediates of formula A and formula B are well known in the art. For example, see Feiser and Feiser, "Reagents for Organic Synthesis", vol. 1, 1967; March, J. "Advanced Organic Chemistry", John Wiley &amp; Sons, 4^to ed, 1992; House, HO, "Modern Synthetic Reactions", 2nd ed., WA Benjamin, New York, 1972; and Larock, RC, Comprehensive Organic Transformations, 2nd Edition, 1999, Wiley-VCH Publishers, New York.

An intermediate of formula A can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0007.tif" />

Chlorination of compound 1 gives compound 2 of chlorine which can be converted to the corresponding amine by treatment with phenol and subsequent reaction with the appropriate amine. The resulting amine can be acylated with the appropriately substituted acyl chloride to provide the intermediate of formula A.

An intermediate of formula B can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0008.tif" />

<img file="ES2311639T3_D0009.tif" />

or

Formula b

ES 2 311 639 T3

Bromination of compound 1 provides compound 3 which can be converted to halogenated compound 4 using procedures known in the art. Reaction with a suitable amine or ammonium salt provides amino compound 5 which can be converted to an intermediate of formula B by treatment with a suitable acid chloride 6.

The following specific values, preferred values, and discussion refer to compounds of formula II.

Specific and preferred values ​​listed below for radicals, substituents, and ranges are for illustration only; they do not exclude other defined values ​​or other values ​​within defined ranges for radicals and substituents.

Specifically, alkyl (C₁ -C₆) can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl or hexyl.

Specifically, alkoxy (C₁ -C₆) can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy or hexoxy.

A specific value for A is CH.

Another specific value for A is N.

A specific value for B is N.

Another specific value for B is CH.

A specific value for W is N.

Another specific value for W is CH.

A specific value for Y is OH.

Another specific value for Y is alkoxy (C₁-C₆).

Another specific value for Y is -OCH3.

Another specific value for Y is alkoxy (C₁ -C₆) replaced.

Another specific value for Y is -OCH2CH2OH.

Another specific value for Y is -OCH2CH2OCH2CH3.

Another specific value for Y is -O-CH2-CHOH-CH2-OH.

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Y is -O-CH2CH2 -NRaRb where Ra and Rb together with the nitrogen to which they are attached form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Y is -OC (= O) CH2-NRaRb.

Another specific value for Y is -OC (= O) -CHOH-CH2-OH.

Another specific value for Y is alkyl (C₁-C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Y is -OC (= O) CH2-NRaRb.

A specific value for Z is OH.

Another specific value for Z is alkoxy (C₁-C₆).

Another specific value for Z is OCH3.

Another specific value for Z is alkoxy (C₁-C₆) replaced.

Another specific value for Z is -OCH₂CH₂OH.

ES 2 311 639 T3

Another specific value for Z is -OCH₂CH₂OCH₂CH₃.

Another specific value for Z is -O-CH₂-CHOH-CH₂ -OH.

Another specific value for Z is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Z is -O-CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Z is -OC (= O) -CHOH-CH2-OH.

Another specific value for Z is alkyl (C₁ -C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Z is -OC (= O) CH2-NRaRb.

A specific value for both R₃ as R₄ it's H.

A specific value for R₃ and R₄ together is = O.

Other specific value for R₃ and R₄ together is = S.

Another specific value for R3 and R4 together is = NH.

Another specific value for R3 and R4 together is = N-R2.

Other specific value for R₃ and R₄ together is = NR₂ in which R₂ is alkyl (C₁-C₆).

Other specific value for R₃ and R₄ together is = NR₂ in which R₂ is alkyl (C₁-C₆) replaced.

A specific value for R₁ it is hydrogen.

Other specific value for R₁ is alkyl (C₁-C₆).

Another specific value for R1 is isobutyl.

Another specific value for R1 is n-butyl.

Another specific value for R1 is isopentyl.

A specific value for R₂ is an alkyl (C₁ -C₆) substituted with one or more hydroxy, mercapto, carboxy, amino, piperazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl groups.

Other specific value for R₂ is an alkyl (C₁ -C₆) with 2 to 4 carbon atoms and substituted with one to two groups selected from hydroxy, mercapto, carboxy, amino, piperazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl.

Other specific value for R₂ is -CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for R2 is -CH2CH2-NRaRb in which Ra and Rb together with the nitrogen to which they are attached form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

A preferred compound of formula (II) is the compound 5-butyl-8,9-dimethoxy-5H-2,3-dioxa-5,10,11,12-tetra-azacyclopenta [b] crisen-6-one; 5-butyl-8,9-dimethoxy-5H-2,3-dioxa-7,10,11,12-tetraaza-cyclopenta [b] crisen-6-one; 5-butyl8,9-dimethoxy-5H-2,3-dioxa-5,7,10,11,12-penta-azacyclopenta [b] crisen-6-one; or a pharmaceutically acceptable salt thereof.

Certain compounds of formula (II) can act as prodrugs for other compounds of formula (II). For example, a compound of formula (II) in which Y and / or Z is -OP (= O) (OH) 2 or -OC (= O) NRcRd; can act as a prodrug for a corresponding compound of formula (II) wherein Y and or Z is hydroxy. Accordingly, a specific subset of compounds of formula (II) are compounds in which Y and / or Z is -OP (= O) (OH) 2 or -OC (= O) NRcRd. A particularly preferred compound is a compound of formula (II) in which Y and / or Z is -OP (= O) (OH) 2. Another preferred compound is a compound of formula (II) in which Y and / or Z is -OC (= O) NRcRd, in which R_c and / or R_d is alkyl (C₁ -C₆) substituted with one or more -NR_andR_F in which R_and and R_F are each independently alkyl (C₁-C₆). Another preferred compound is a compound of formula (II) in which Y and / or Z is -OC (= O) NR_cR_d, in which R_c and R_d together with the nitrogen to which they are attached they form an N '- (alkyl) piperazino, pyrrolidino or piperidino ring. A more preferred compound is a compound of formula (II) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd

ES 2 311 639 T3 together with the nitrogen to which they are attached form a piperidinyl ring, which ring is optionally substituted with an N-linked heterocycle ring (eg piperidino).

The present invention provides compounds of formula II and a process for preparing compounds of formula II wherein R₁ is such as alkyl (C₁-C₆) or alkyl (C₁-C₆) substituted which comprises reacting the compound of formula II wherein R1 is H with a suitable nitrogen alkylating agent, such as a (C1-C6) alkyl halide or substituted (C1-C6) alkyl halide, to form a corresponding substituted (C1-C6) alkyl or (C1-C6) alkyl compound. One skilled in the art understands that the nitrogen atom of lactam can be conveniently synthetically manipulated and efficiently converted into useful related compounds, for example, by preparing intermediates with a protected N atom and whose protected nitrogen atom can be deprotected and subsequently alkylated for providing the aforementioned alkylated nitrogen compounds.

A compound of formula II can be prepared by subjecting a corresponding intermediate of formula A to suitable cycling conditions; for example, by treatment with palladium acetate and tri-o-tolylphosphine, as illustrated in Scheme 1 below. A compound of formula II can be prepared by subjecting a corresponding intermediate of formula B to conditions suitable for the formation of the tetracyclic ring system; for example, by treatment with a suitable tin reagent, as illustrated in Scheme 2 below.

Scheme 1

<img file="ES2311639T3_D0010.tif" />

X

Formula A Formula II

Scheme 2

<img file="ES2311639T3_D0011.tif" />

Formula B Formula II

Other suitable conditions for the formation of the ring system from intermediates of formula A and formula B are well known in the art. For example, see Feiser and Feiser, "Reagents for Organic Synthesis", vol. 1, 1967; March, J. "Advanced Organic Chemistry", John Wiley &amp; Sons, 4^to ed., 1992; House, HO, "Modern Synthetic Reactions", 2nd ed., WA Benjamin, New York, 1972; and Larock, RC, Comprehensive Organic Transformations, 2nd ed., 1999, Wiley-VCH Publishers, New York.

An intermediate of formula A can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

ES 2 311 639 T3

<img file="ES2311639T3_D0012.tif" />

Similarly, an intermediate of formula B can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0013.tif" />

An alternative route for the formation of 5,6-dihydro derivatives of formula II involves the reduction of the lactam or the desulfurization of the thioamide as illustrated by the following. Additionally, compounds of formula II can be modified to provide other compounds of formula II as illustrated below.

<img file="ES2311639T3_D0014.tif" />

where X = O or S

<img file="ES2311639T3_D0015.tif" />

where X = O or S

ES 2 311 639 T3

<img file="ES2311639T3_D0016.tif" />

where X = S, NR₂

The following specific values, preferred values, and discussion refer to compounds of formula III.

Specifically, alkyl (C -C₆) can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl or hexyl.

Specifically, alkoxy (C -C₆) can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy or hexoxy.

A specific value for X is = 0.

Another specific value for X is = S.

Another specific value for X is = NH.

Another specific value for X is = N-R2.

Another specific value for X is = NR₂ in which R₂ is alkyl (C₁-C₆).

Another specific value for X is = NR₂ in which R₂ is alkyl (C₁-C₆) replaced.

A specific value for Y is OH.

Another specific value for Y is alkoxy (C₁-C₆).

Another specific value for Y is -OCH₃.

Another specific value for Y is alkoxy (C₁ -C₆) replaced.

Another specific value for Y is -OCH₂CH₂OH.

Another specific value for Y is -OCH₂CH₂OCH₂CH₃.

Another specific value for Y is -O-CH₂-CHOH-CH₂-OH.

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Y is -OC (= O) CH₂-NR_toR_b.

Another specific value for Y is -OC (= O) -CHOH-CH₂-OH.

Another specific value for Y is alkyl (C₁-C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Y is -OC (= O) CH₂-NR_toR_b.

A specific value for Z is OH.

Another specific value for Z is alkoxy (C₁-C₆).

Another specific value for Z is OCH₃.

Another specific value for Z is alkoxy (C₁-C₆) replaced.

Another specific value for Z is -OCH₂CH₂OH.

ES 2 311 639 T3

Another specific value for Z is -OCH2CH2OCH2CH3.

Another specific value for Z is -O-CH₂-CHOH-CH₂ -OH.

Another specific value for Z is -O-CH2CH2-NRaRb where Ra and Rb are hydrogen or (C1-C6) alkyl.

Another specific value for Z is -O-CH2CH2-NRaRb where Ra and Rb together with the nitrogen to which they are attached form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Z is -OC (= O) -CHOH-CH2-OH.

Another specific value for Z is (C1-C6) alkyl substituted with one or more tetrahydrofuranyl, tetrahydropyranyl, or 1,4-dioxanyl rings.

Another specific value for Z is -OC (= O) CH2-NRaRb.

A specific value for R1 is methyl, ethyl, propyl, or isopropyl.

A preferred compound of formula (III) is the compound 5-butyl-8,9-dimethoxy-5H-2,3-dioxa-5,11,12-triaza-cyclopenta [b] crisen-6-one, 5-isobutyl -8,9-dimethoxy-5H-2,3-dioxa-5,11,12-triaza-cyclopenta [b] crisen-6-one, or a pharmaceutically acceptable salt thereof.

Certain compounds of formula (III) can act as prodrugs for other compounds of formula (III). For example, a compound of formula (III) in which Y and / or Z is -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; can act as a prodrug for a corresponding compound of formula (III) in which Y and or Z is hydroxy. Accordingly, a specific subset of compounds of formula (III) are compounds in which Y and / or Z is -OP (= O) (OH) 2 or -OC (= O) NRcRd. A particularly preferred compound is a compound of formula (III) in which Y and / or Z is -OP (= O) (OH) 2. Another preferred compound is a compound of formula (III) in which Y and / or Z is -OC (= O) NRcRd, in which R_c and / or R_d is alkyl (C₁ -C₆) substituted with one or more -NR_andR_F in which R_and and R_F are each independently alkyl (C₁-C₆). Another preferred compound is a compound of formula (III) in which Y and / or Z is -OC (-O) NR_cR_d, wherein Rc and Rd together with the nitrogen to which they are attached form an N '- (alkyl) piperazino, pyrrolidino or piperidino ring. A more preferred compound is a compound of formula (III) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd together with the nitrogen to which they are attached form a piperidinyl ring, which ring it is optionally substituted with an N-linked heterocycle ring (eg piperidino).

The present invention provides compounds of formula III and a process for preparing compounds of formula III wherein R₁ is such as alkyl (C₁-C₆) or alkyl (C₁ -C₆) substituted which comprises reacting the compound of formula I wherein R1 is H with a suitable nitrogen alkylating agent, such as an alkyl halide (C₁ -C₆) or alkyl halide (C₁-C₆) substituted, to form an alkyl compound (C₁-C₆) or alkyl (C₁C6) substituted corresponding. One skilled in the art understands that the nitrogen atom of lactam can be conveniently synthetically manipulated and efficiently converted into useful related compounds, for example, by preparing intermediates with a protected N atom and whose protected nitrogen atom can be deprotected and subsequently alkylated for providing the aforementioned alkylated nitrogen compounds.

A compound of formula III can be prepared by subjecting a corresponding intermediate of formula A to suitable cycling conditions; for example, by treatment with palladium acetate and tri-o-tolylphosphine, as illustrated in Scheme 1 below. A compound of formula III can also be prepared by subjecting a corresponding intermediate of formula B to conditions suitable for the formation of the ring system; for example by treatment with a suitable tin reagent, as illustrated in Scheme 2 below. The compounds of the present invention include intermediates of formulas A and B.

Scheme I

<img file="ES2311639T3_D0017.tif" />

ES 2 311 639 T3

Scheme 2

<img file="ES2311639T3_D0018.tif" />

Other suitable conditions for the formation of the ring system from intermediates of formula A and formula B are well known in the art. For example, see Feiser and Feiser, "Reagents for Organic Synthesis", vol. 1, 1967; March, J. "Advanced Organic Chemistry", John Wiley &amp; Sons, 4th ed., 1992; House, HO, "Modern Synthetic Reactions", 2nd ed., WA Benjamin, New York, 1972; and Larock, RC, Comprehensive Organic Transformations, 2nd ed., 1999, Wiley-VCH Publishers, New York.

An intermediate of formula A can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0019.tif" />

Similarly, an intermediate of formula B can be prepared from readily available starting materials using procedures that are known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0020.tif" />

Alternatively the compounds of formula III can be modified to form related derivatives of formula III as illustrated by the following.

ES 2 311 639 T3

<img file="ES2311639T3_D0021.tif" />

where X = O or S

<img file="ES2311639T3_D0022.tif" />

where X = S, NR2

The following specific values, preferred values, and discussion refer to compounds of formula IV.

Specifically, alkyl (C₁ -C₆) can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl or hexyl.

Specifically, alkoxy (C₁ -C₆) can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy or hexoxy.

A specific compound of formula IV is a compound of formula XX:

<img file="ES2311639T3_D0023.tif" />

Another specific compound of formula IV is a compound of formula XXI:

<img file="ES2311639T3_D0024.tif" />

A specific value for W is N. Another specific value for W is CH.

A specific value for A is CH.

Another specific value for A is N.

A specific value for B is N. Another specific value for B is CH.

ES 2 311 639 T3

A specific value for Yes OH.

Another specific value for Y is alkoxy (C₁-C₆).

Another specific value for Y is -OCH3.

Another specific value for Y is alkoxy (C₁ -C₆) replaced.

Another specific value for Y is -OCH2CH2OH.

Another specific value for Y is -OCH2CH2OCH2CH3.

Another specific value for Y is -O-CH2-CHOH-CH2-OH.

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Y is -O-CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Y is -OC (= O) CH2-NRaRb,

Another specific value for Y is -OC (= O) -CHOH-CH2-OH.

Another specific value for Y is alkyl (C₁-C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Y is -OC (= O) CH2-NRaRb.

A specific value for Z is OH.

Another specific value for Z is alkoxy (C₁-C₆).

Another specific value for Z is OCH3.

Another specific value for Z is alkoxy (C₁-C₆) replaced.

Another specific value for Z is -OCH₂CH₂OH.

Another specific value for Z is -OCH₂CH₂OCH₂CH₃.

Another specific value for Z is -O-CH₂-CHOH-CH₂ -OH.

Another specific value for Z is -O-CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Another specific value for Z is -O-CH2CH2-NRaRb where Ra and Rb together with the nitrogen to which they are attached form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for Z is -OC (= O) -CHOH-CH2-OH.

Another specific value for Z is alkyl (C₁ -C₆) substituted with one or more tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl rings.

Another specific value for Z is -OC (= O) CH2-NRaRb.

A specific value for R₁ or R₂ is an alkyl (C₁-C₆) substituted with one or more hydroxy groups.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with a hydroxy group.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with one or more mercapto groups.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with a mercapto group.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with one or more carboxy groups.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with a carboxy group.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with one or more NR groups_toR_b.

ES 2 311 639 T3

Other specific value for R₁ or R₂ is an alkyl (C₁-C₆) substituted with an NR group_toR_b.

Other specific value for R₁ or R₂ is an alkyl (C₁-C₆) substituted with one or more NH groups₂.

Other specific value for R₁ or R₂ is an alkyl (C₁-C₆) substituted with an NH group₂.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) substituted with one or more hydroxy, mercapto, carboxy, amino, piperazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl groups.

Other specific value for R₁ or R₂ is an alkyl (C₁ -C₆) with 2 to 4 carbon atoms and substituted with one to two groups selected from hydroxy, mercapto, carboxy, amino, piperazinyl, pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl or 1,4-dioxanyl.

Another specific value for R1 or R2 is 2-hydroxymethyl.

Another specific value for R1 or R2 is 2-hydroxyethyl.

Another specific value for R1 or R2 is 3-hydroxypropyl.

Another specific value for R1 or R2 is 2-hydroxypropyl.

Another specific value for R1 or R2 is H.

Another specific value for R1 or R2 is -CH2-CHOH-CH2-OH.

Other specific value for R₁ or R₂ is -CH₂CH₂-NR_toR_b in which R_to and R_b are hydrogen or alkyl (C₁-C₆).

Other specific value for R₁ or R₂ is -CH₂CH₂-NR_toR_b in which R_to and R_b together with the nitrogen to which they are attached they form a piperazino, pyrrolidino, piperidino, morpholino or thiomorpholino ring.

Another specific value for R1 and R2 together is = 0.

Another specific value for R1 and R2 together is = S.

Another specific value for R1 and R2 together is = NH.

A preferred compound of formula (IV) is the compound 8,9-dimethoxy-6H-2,3,5-trioxa-11,12-diaza-cyclopenta [b] chrysen, 8,9-dimethoxy-6H-2,3 , 5-trioxa-12-aza-cyclopenta [b] chrysen, or a pharmaceutically acceptable salt thereof.

Certain compounds of formula (IV) can act as prodrugs for other compounds of formula (IV). For example, a compound of formula (IV) in which Y and / or Z is -OP (= O) (OH)₂ or -OC (= O) NR_cR_d; can act as 1 prodrug for a corresponding compound of formula (IV) wherein Y and or Z is hydroxy. Accordingly, a specific subset of compounds of formula (IV) are compounds in which Y and / or Z is -OP (= O) (OH) 2 or -OC (= O) NRcRd. A particularly preferred compound is a compound of formula (IV) in which Y and / or Z is -OP (= O) (OR) 2. Another preferred compound is a compound of formula (IV) in which Y and / or Z is -OC (= O) NR_cR_d, in which R_c and / or R_d is alkyl (C₁ -C₆) substituted with one or more -NR_andR_F in which R_and and R_F are each independently alkyl (C₁-C₆). Another preferred compound is a compound of formula (IV) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd together with the nitrogen to which they are attached form an N '- (alkyl ) piperazino, pyrrolidino or piperidino. A more preferred compound is a compound of formula (IV) in which Y and / or Z is -OC (= O) NRcRd, in which Rc and Rd together with the nitrogen to which they are attached form a piperidinyl ring, which ring it is optionally substituted with an N-linked heterocycle ring (eg piperidino).

A compound of formula IV can be prepared by subjecting a corresponding intermediate of formula A to suitable cycling conditions; for example, by treatment with palladium acetate and tri-o-tolylphosphine, as illustrated in Scheme 1 below. A compound of formula IV can also be prepared by subjecting a corresponding intermediate of formula B to conditions suitable for the formation of the ring system; for example by treatment with a suitable tin reagent 1, as illustrated in Scheme 2 below. The compounds of the present invention include intermediates of formulas A and B.

ES 2 311 639 T3

Scheme 1

<img file="ES2311639T3_D0025.tif" />

Scheme 2

<img file="ES2311639T3_D0026.tif" />

Other suitable conditions for the formation of the ring system from intermediates of formula A and formula B are well known in the art. For example, see Feiser and Feiser, "Reagents for Organic Synthesis", vol. 1, 1967; March, J. "Advanced Organic Chemistry", John Wiley &amp; Sons, 4th ed, 1992; House, HO, "Modern Synthetic Reactions", 2nd ed., WA Benjamin, New York, 1972; and Larock, RC, Comprehensive Organic Transformations, 2nd Edition, 1999, Wiley-VCH Publishers, New York.

An intermediate of formula A can be prepared from readily available starting materials using procedures known in the art, or the procedures illustrated below can be prepared.

<img file="ES2311639T3_D0027.tif" />

Similarly, an intermediate of formula B can be prepared from readily available starting materials using procedures known in the art, or it can be prepared using the procedures illustrated below.

<img file="ES2311639T3_D0028.tif" />

ES 2 311 639 T3

<img file="ES2311639T3_D0029.tif" />

Rl ^r2

Formula Θ

Alternatively the compounds of formula IV can be modified to form desired derivatives related to formula IV as illustrated below.

<img file="ES2311639T3_D0030.tif" />

where R2 is not H

<img file="ES2311639T3_D0031.tif" />

where R1 is not H

<img file="ES2311639T3_D0032.tif" />

where X = S, NR2

General discussion

The starting materials used in the synthetic procedures described herein are commercially available, have been described in the scientific literature, or can be prepared from readily available starting materials using procedures known in the field. It may be desired to optionally use a protecting group during all or part of the synthetic procedures described above. Such protecting groups and procedures for their introduction and removal are well known in the art. See Greene, TW; Wutz, PGM "Protecting Groups In Organic Synthesis" Second Edition, 1991, New York, John Wiley &amp; Sons, Inc.

It will be appreciated by those skilled in the art that compounds of the invention having a chiral center can exist in and be isolated in optically active and racemic forms. Some compounds may have polymorphism. It should be understood that the present invention encompasses any racemic, optically active, polymorphic, or stereoisomeric form, or mixtures thereof, of a compound of the invention, which possesses the useful properties described herein, it being well known in the art how to prepare forms optically active (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis or by chromatographic separation using a chiral stationary phase) and how to determine topoisomerase inhibition activity or cytotoxic activity using the standard tests described herein, or using other similar tests that are well known in the art. The compounds of the present invention may contain chiral centers, for example, in any of the Y, Z and R substituents.₁.

ES 2 311 639 T3

In cases where the compounds are sufficiently basic or acidic to form stable non-toxic acid or base salts, administration of the compounds as salts may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids that form a physiologically acceptable anion, for example tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, α-ketoglutarate and α-glycerophosphate . Suitable inorganic salts can also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.

Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound such as an amine with a suitable acid that provides a physiologically acceptable anion. With an alkali metal, for example sodium, potassium or lithium, or alkaline earth metal, for example calcium, salts of carboxylic acids can also be prepared.

The compounds of the invention can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms adapted to the chosen route of administration, that is, orally or parenterally, intravenously, intramuscular, topical or subcutaneous.

Thus, the present compounds can be administered systemically, for example orally, in combination with a pharmaceutically acceptable carrier such as an inert diluent or an assimilable edible carrier. They can be encased in hard or soft-shell gelatin capsules, can be compressed into tablets, or can be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound can be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, thick tablets, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations must contain at least 0.1% of the active compound. Of course, the percentage of the compositions and preparations can be varied and can conveniently be between about 2 and about 60% by weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

Tablets, coarse tablets, pills, capsules and the like may also contain the following: binders such as gum tragacanth, acacia, cornstarch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid, and the like; a lubricant such as magnesium stearate; and a sweetener such as sucrose, fructose, lactose or aspartame or a flavoring such as peppermint, wintergreen oil or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills or capsules can be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir can contain the active compound, sucrose or fructose as a sweetener, methyl and propyl parabens as preservatives, a coloring and flavoring such as cherry or orange flavoring. Of course, any material used in the preparation of any unit dosage form must be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound can be incorporated into sustained-release preparations and devices.

The active compound can also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a non-toxic surfactant. Dispersions can also be prepared in glycerin, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under normal storage and use conditions, these preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient that are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final pharmaceutical form must be sterile, fluid and stable under the conditions of preparation and storage. The liquid carrier or vehicle may be a liquid solvent or dispersion medium comprising, for example, water, ethanol, a polyol (eg, glycerin, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, non-toxic glyceryl esters, and mixtures. adequate of them. The proper fluidity can be maintained, for example, by the formation of liposomes, by maintaining the required particle size in the case of dispersions or by the use of surfactants. Prevention of the action of microorganisms can be accomplished by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases it will be preferred to include isotonic agents, for example sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents that delay absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other components listed above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation procedures are vacuum drying and lyophilization techniques, which yield a powder of the active ingredient plus any additional desired components present in the previously sterile filtered solutions. .

ES 2 311 639 T3

For topical administration, the present compounds can be applied neat, that is, when they are liquid. However, it will generally be desired to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which can be a solid or a liquid.

Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina, and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol / glycol mixtures in which the present compounds can be dissolved or dispersed to effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize properties for a given use. The resulting liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using aerosol or pump type sprays.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or mineral materials modified with liquid carriers can also be employed to form spreads, gels, ointments, soaps and the like, for direct application to the user's skin.

Examples of useful dermatological compositions that can be used to administer the compounds of the invention to the skin are known in the art; for example, see Jacquet et al. (US Patent No. 4,608,392), Geria (US Patent No. 4,992,478), Smith et al. (US Patent No. 4,559,157) and Wortzman (US Patent No. 4,820,508).

Useful dosages of the compounds of the invention can be determined by comparing their in vitro activity and in vivo activity in animal models. Procedures for extrapolation of effective dosages in mice, and other animals, to humans are known in the art; for example, see US Patent No. 4,938,949.

Generally, the concentration of the compound (s) of the invention in a liquid composition, such as a lotion, will be about 0.1-25% by weight, preferably about 0.5-10% by weight. weight. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.15% by weight, preferably about 0.5-2.5% by weight.

The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected, but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient. patient and will be ultimately at the discretion of the practitioner or clinician.

In general, however, a suitable dose will be in the range of about 0.5 to about 100 mg / kg, for example, about 10 to about 75 mg / kg of body weight per day, such as 3 to about 50 mg per kilogram of recipient body weight per day, preferably in the range of 6 to 90 mg / kg / day, most preferably in the range of 15 to 60 mg / kg / day.

The compound can conveniently be administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently 50 to 500 mg of active principle per unit dosage form.

Ideally, the active ingredient should be administered to achieve peak plasma concentrations of the active compound of about 0.5 to about 75 µΜ, preferably about 1 to 50 µΜ, most preferably about 2 to about 30 µ. This can be achieved, for example, by intravenous injection of a 0.05 to 5% solution of the active ingredient, optionally in saline, or administered orally as a bolus containing about 1-100 mg of the active ingredient. Desirable blood levels can be maintained by continuous infusion to provide about 0.01-5.0 mg / kg / h or by intermittent infusions containing about 0.4-15 mg / kg of the active ingredient (s). ).

The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, eg, in two, three, four or more sub-doses per day. The subdose itself can be further divided, for example, into several freely separated discrete administrations; such as multiple inhalations of an insufflator or by applying a plurality of drops to the eye.

The ability of a compound of the invention to effect topoisomerase I or II mediated DNA cleavage can be determined using pharmacological models that are well known in the art, for example, using a model such as Test A described below.

Test A

Topoisomerase I-mediated DNA cleavage assay

Human topoisomerase I was expressed in E. Coli and isolated as a recombinant fusion protein using a T7 expression system as previously described, see Makhey, D. et al., Bioorg. Med. Chem., 2000, 8, 111. DNA topoisomerase I was purified from the calf thymus gland as previously reported, see Maniatis, T., et al., J. Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor,

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New York, 149-185). The YepG plasmid was also purified by the alkali lysis procedure followed by phenol deproteinization and the isopycnic CsCl / ethidium centrifugation procedure as described, see Maniatis, T .; Fritsch, EF; Sambrook, J. Molecular Cloning, a Laboratory Manual; Cold Spring Harbor Laboratory: Cold Spring Harbor, NY 1982; P. 149-185. End labeling of the plasmid was performed by restriction enzyme digestion followed by end filling with Kienow polymerase as previously described, see Liu, LF; Rowe, TC; Yang, L .; Tewey, KM; Chen, GL, J. Biol. Chem. 1983, 258, 15365. Cleavage assays were performed as previously reported, see B. Gatto et al. Cancer Res., 1996, 56, 2795-2800. The drug and DNA in the presence of topoisomerase I were incubated for 30 minutes at 37 ° C. After development of the gels, a 24 hour exposure was typically used to obtain autoradiograms that gave a general idea of ​​the degree of DNA fragmentation. Topoisomerase I-mediated DNA cleavage values ​​are reported as CER, relative effective concentration, that is, concentrations relative to 2,3-dimethoxy-8,9-methylenedioxybenzo [i] phenanthridine, arbitrarily assumed to be 1.0 , which can produce the same cleavage in plasmid DNA in the presence of human topoisomerase I. Relative potency was based on the relative amount of drug needed to induce approximately 10% DNA fragmentation. The trials are conducted under the direction of Dr. LF Liu, Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ.

A similar assay can be used to assess the ability of a compound of the invention to effect topoisomerase II mediated DNA cleavage by replacing the human topoisomerase I used in Test A with a suitable topoisomerase II.

The cytotoxic effects of a compound of the invention can be determined using pharmacological models that are well known in the art, for example, using a model such as Test B described below.

Test B

Cell growth inhibition: MTT microtiter plate tetrazolinium cytotoxicity assay (RPMI 8402, CPT-K5, U937, U937 / CR cells)

Cytotoxicity is determined using the MTT Microtiter Plate Tetrazolinium Cytotoxicity Assay (MTA), see Chen AY et al. Cancer Res. 1993, 53, 1332; Mosmann, TJ, J. Immunol. Methods 1983, 65, 55; and Carmichael, J. et al. Cancer Res. 1987,47,936. Human lymphoblast RPMI 8402 and its camptothecin resistant variant cell line CPT-K5 were provided by Dr. Toshiwo Andoh (Anchi Cancer Research Institute, Nagoya, Japan), see Andoh, T .; Okada, K, Adv. in Pharmacology 1994, 29B, 93. Human U-937 myeloid leukemia cells and U-937 / CR cells were described by Rubin et al., J. Biol. Chem., 1994, 269, 2433-2439. The cytotoxicity assay is performed using 96-well microtiter plates using 2000 cells / well in 200 ml of growth medium. Cells are grown in suspension at 37 ° C in 5% CO₂ and they are maintained by regular passage in RPMI medium supplemented with 10% heat inactivated fetal bovine serum, L-glutamine (2 mM), penicillin (100 U / ml) and streptomycin (0.1 mg / ml). For the determination of CI₅₀, cells are continuously exposed for 3-4 days to varying drug concentrations and MU assays were performed at the end of the fourth day. Each test is performed with a control that did not contain drug. All assays are performed at least twice in 6 duplicate wells. All trials are conducted under the direction of Dr. LF Liu, Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ.

Compounds of the invention can act as cytotoxic agents against tumor cell lines including multidrug resistant tumor cell lines. Thus, the compounds are useful for treating cancer and can be used to treat tumors that are resistant to other specific chemotherapeutic agents.

Topoisomerase inhibitors are also known to possess antibacterial, antifungal, antipsoriatic (psoriasis), antiprotozoal, anthelmintic, and antiviral activity. Accordingly, the topoisomerase inhibitors of the invention may also be useful as antibacterial, antifungal, antipsoriatic (psoriasis), antiprotozoal, anthelmintic or antiviral agents. In particular, the compounds of the invention that demonstrate little or no activity as mammalian topoisomerase I poisons, due to the possibility of a similar molecular mechanism of action, could be antibacterial, antifungal, antipsoriatic (psoriasis), antiprotozoal, anthelmintic or highly active and selective antivirals. Thus, certain compounds of the invention may be particularly useful as antibacterial, antifungal, antipsoriatic (psoriasis), antiprotozoal, anthelmintic, or systemic antiviral agents in mammals. The invention also provides the use of a compound of the invention for the preparation of a medicament useful for producing an antibacterial, antifungal, antipsoriatic (psoriasis), antiprotozoal, anthelmintic or antiviral effect in a mammal.

As used herein, the term "solid mammalian tumors" includes cancers of the head and neck, lung, mesothelioma, mediastinum, esophagus, stomach, pancreas, hepatobiliary system, small intestine, colon, rectum, anus, kidney, ureter, bladder, prostate, urethra, penis, testicle, gynecological organs, ovarian, breast, endocrine system, skin, central nervous system; soft tissue and bone sarcomas; and melanoma of cutaneous and intraocular origin. The term "hematologic malignancies" includes childhood leukemia and lymphomas, Hodgkin's disease, lymphomas of lymphocytic and cutaneous origin, acute and chronic leukemia, plasma cell neoplasia, and AIDS-associated cancers. The preferred mammalian species for treatment are humans and domesticated animals.

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The invention will now be illustrated by the following non-limiting examples. Specific compounds of the present invention can be prepared as illustrated in the following schemes using known reactive reactions.

Example 1

Preparation of representative compounds of formula I

<img file="ES2311639T3_D0033.tif" />

Example 2

Preparation of representative compounds of formula I

<img file="ES2311639T3_D0034.tif" />

Specific compounds of the present invention can be prepared according to the following schemes using known reactions and reagents.

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Example 3

Preparation of representative compounds of formula II

<img file="ES2311639T3_D0035.tif" />

<img file="ES2311639T3_D0036.tif" />

Example 4

Preparation of representative compounds of formula II

<img file="ES2311639T3_D0037.tif" />

<img file="ES2311639T3_D0038.tif" />

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Example 5

Preparation of representative compounds of formula III

Example 6

<img file="ES2311639T3_D0039.tif" />

<img file="ES2311639T3_D0040.tif" />

Preparation of representative compounds of formula III

<img file="ES2311639T3_D0041.tif" />

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Example 7

Preparation of representative compounds of formula IV

<img file="ES2311639T3_D0042.tif" />

Example 8

Synthesis of 8,9-dimethoxy-2,3-methylenedioxy-5- (n-butyl) -5H-5,11,12-triazacrisen-6-one

A mixture of N- (6,7-methylenedioxycinolin-4-yl) -N- (n-butyl) -2-iodo-4,5-dimethoxybenzamide (1.0 equiv. In mmol), Pd (OAc)₂ (0.2 mmol equiv.), P (o-tolyl)₃ (0.4 equiv. In mmol) and Ag₂CO₃ (2.0 mmol equiv) was refluxed in DMF (30 ml per mmol equiv) with stirring. The reaction mixture was allowed to cool to room temperature, diluted with CHCl₃ and leaked through Celite. The dried product was washed extensively with 10% CH₃OH in CHCl₃. The filtrate was concentrated in vacuo and the residue was chromatographed on silica gel using chloroform: methanol to provide the title compound (123 mg, 0.2 mmol) in 27% yield with a reaction time of 90 min; mp. 299 ° C; IR (KBr) 1654; NMR¹H (CDCl3) δ 1.06 (t, 3H, J = 7.4), 1.56 (m, 2H), 2.13 (m, 2H), 4.09 (s, 3H), 4.17 (s, 3H), 4.49 (m, 2H), 6.26 (s, 2H), 7.62 (s, 1H), 7.85 (s, 1H), 7.87 (s, 1H) , 8.65 (s, 1H); NMR¹³C (CDCl3) δ 13.8, 20.2, 31.1, 48.6, 56.3, 56.6, 98.8, 102.7, 104.2, 106.5, 107.9, 119.7 , 149.1, 150.1, 150.9, 151.4, 153.6, 154.2, 162.9; HRMS calcd for C₂₂H₂₁N₃OR₅H: 408.1559; found 408.1543.

The intermediate compound N- (6,7-methylenedioxycinolin-4-yl) -N- (n-butyl) -2-iodo-4,5-dimethoxybenzamide was prepared as follows.

to. 6,7-Methylenedioxy-4-cinnolone. A mixture of 6'-amino-3 ', 4'-methylenedioxyacetophenone (2.4 g, 13.4 mmol) in concentrated hydrochloric acid (92 ml) and water (13 ml) was cooled to -5 ° C and diazotized by dropwise addition of a solution of sodium nitrite (0.925 g, 13.4 mmol) in water (4 ml). After stirring for an additional hour at -5 ° C, the mixture was transferred to a bath preheated to 75 ° C and left stirring at this temperature overnight. The reaction mixture was cooled to 5 ° C to induce crystallization. This material was filtered and then added to 10% aqueous NaOH (100 ml), which was again filtered and dried in vacuo to give 2.37 g of the cinnoline as a colorless solid in 93% yield; mp. 318-320 ° C; NMR¹H (DMSO-d6) δ 6.21 (s, 2H), 6.97 (s, 1H), 7.30 (s, 1H), 7.63 (s, 1H); NMR¹³C (DMSOd6) δ 94.9, 100.3, 103.3, 120.1, 139.7, 139.9, 147.4, 153.5, 169.4; HRMS calcd for C₉H6O3N2: 190.0378; found: 190.0372.

ES 2 311 639 T3

b. 4-Chloro-6,7-methylenedioxycinoline. 6,7-methylenedioxy-4-cinnolone (1.0 g, 5.3 mmol) was added in small portions to a stirred mixture of phosphorus pentachloride (1.4 g, 6.7 mmol) and phosphorus oxychloride ( 4 ml, 6.6 mmol) at room temperature. The reaction flask was heated to 80 ° C for 4 hours, then cooled to room temperature and poured onto 50 g of crushed ice. After neutralization of the solution with solid sodium acetate, the precipitate was removed by filtration and recrystallized from ethanol to give 800 mg of the chlorocinoline as an off-white solid in 73% yield; mp. 203.5204.5 ° C; NMR¹H (CDCl3) δ 6.25 (s, 2H), 7.39 (s, 1H), 7.73 (s, 1H), 9.14 (s, 1H); NMR¹³C (CDCl3) δ

97.8, 102.9, 105.1, 124.2, 133.4, 144.0, 150.0, 152.3, 152.7; HRMS calcd for C9H5O2N2Q: 208.0040; found: 208.0042.

c. N- (6,7-Methylenedioxycinolin-4-yl) -n-butylamine. Butylamine ((25 ml)) was added with stirring to 4-chloro-6,7 methylenedioxycinoline (1 g, 4.7 mmol) and copper powder (250 mg). The reaction was then allowed to stir for 18 h at 80 ° C and the solvent was removed under reduced pressure. The residue was partitioned between CHCl₃and 10% NaOH. The aqueous phase was repeatedly separated with CHCl₃. All CHCl solutions₃(initial partition and extracts) were combined and dried (MgSO4) to provide the product in 32.5% yield; mp. 247-248 ° C; NMR¹H (CDCl3) δ 1.02 (t, 3H, J = 7.4), 1.50 (m, 2H), 1.73 (m, 2H), 3.40 (m, 2H), 4.59 (s, 1H), 6.14 (s, 2H), 6.96 (s, 1H), 7.57 (s, 1H), 8.59 (s, 1H); NMR¹³C (CDCl3) δ 13.8; 20.2,

29.7, 31.3, 42.8, 5 94.2, 102.1, 105.3, 112.7, 126.7, 140.6, 149.6, 150.7; HRMS calcd for C₁₃H₁₅N₃OR₂H: 246.1243; found 246.1237.

d. N- (6,7-Methylenedioxycinolin-4-yl) -N- (n-butyl) -2-iodo-4,5-dimethoxybenzamide. A 2.0 M solution of oxalyl chloride in CH₂Cl₂ (1.3 equiv.) Was added to a solution of 2-iodo-4,5-dimethoxybenzoic acid (1.0 equiv.) In CH₂Cl₂ anhydrous (~ 60 ml per 10 mmol benzoic acid) and the solution was stirred under reflux for 3 h. The mixture was allowed to cool and then concentrated to dryness in vacuo. To the residue was added a solution of the appropriate N- (6,7-methylenedioxycinolin-4-yl) -n-butylamine (1.0 equiv.), Triethylamine (2 equiv.) In CH₂Cl₂ (~ 60 ml per 4 mmol aminoquinoline). Then the reaction mixture was stirred under reflux under N2. The reaction mixture was cooled and washed with sat. NaHCO3. and extracted with 3% HCl. The aqueous phase was neutralized with 20% NaOH and extracted with CHCl₃, dried (MgSO₄) and evaporated to provide the product (350 mg, 1.4 mmol); in 19% yield with a reaction time of 18 h at 50 ° C from the acid chloride prepared using 5.0 mmol of oxalyl chloride and 2.1 mmol of 2-iodo-4,5-dimethoxybenzoic acid; mp. 133-134 ° C; IR (KBr) 1654; NMR¹H (CDCl3) δ 0.87 (t, 3H, J = 7.2), 1.20-1.90 (m, 4H), 3.33 (s, 3H), 3.68 (s, 3H) , 3.90 (m, 1H), 4.35 (m, 1H), 6.19 (d, 2H, J = 3.2), 6.34 (s, 1H), 6.98 (s, 1H), 7.25 (s, 1H), 7.62 (s, 1H), 9.01 (s, 1H); NMR¹³C (CDCl3) δ 13.7, 20.1, 3 0, 0, 49.4, 55.7, 56.1, 82, 9, 96, 5, 102, 9, 105.6, 110, 5, 121.9, 133.1, 148.3, 150.0, 151.8, 152.5, 169.7; HRMS calcd for C22H22IN3O₅Li 542.0764; found 542.0757.

Example 9

Synthesis of 2,3-dimethoxy-8,9-methylenedioxy-5- (butyl) -5H-dibenzo [c₂hJ1,6-naphthyridin-6-one

A mixture of N- (6,7-methylenedioxyquinolin-4-yl) -N- (butyl) -2-iodo-4,5-dimethoxybenzamide (1.0 equiv. In mmol), Pd (OAc) 2 (0 , 2 mmol equiv), P (o-tolyl) 3 (0.4 mmol equiv) and Ag2CO3 (2.0 mmol equiv) was refluxed in DMF (30 ml per mmol equiv) with agitation. The reaction mixture was allowed to cool to room temperature, diluted with CHCl₃ and leaked through Celite. The dried product was washed extensively with 10% CH3OH in CHCl3. The filtrate was concentrated in vacuo and the residue was chromatographed on silica gel using chloroform: methanol to provide the title compound; (24% yield); reaction time 45 min; mp. 224 ° C (dec.); IR (KBr) 1654; NMR¹H (CDCl3); δ 0.99 (t, 3H, J = 7.4), 1.62 (m, 2H), 2.09 (m, 2H), 4.07 (s, 3H), 4.14 (s, 3H ), 4.49 (m, 2H), 6.19 (s, 2H), 7.50 (s, 1H), 7.61 (s, 1H), 7.70 (s, 1H), 7.92 (s, 1H), 9.40 (s, 1H); NMR¹³C (CDCl3) δ 13.7, 20.2, 31.2, 50.6, 56.3, 56.4, 100.7, 102.0, 102.2, 107.4, 108.8, 111 , 7, 114.9, 119.5, 127.4, 141.1, 143.7, 147.1, 147, 5, 149, 8, 150, 3, 154.1,164.0; HRMS calcd for C43H22N2O5H: 406.1529; found 406.1534.

The intermediate N- (6,7-methylenedioxyquinolin-4-yl) -N- (butyl) -2-iodo-4,5-dimethoxybenzamide was prepared as follows.

to. 4-Chloro-6,7-methylenedioxyquinoline. It was prepared from 4-hydroxy-6,7-methylenedioxyquinoline using procedures previously described in the literature for the conversion of 4-hydroxyquinoline to 4-chloroquinoline. Compound 5 had: mp. 127.5-128 ° C (Bibl.²⁸ mp. 129 ° C); NMR¹H (CDCl₃) 6.15 (s, 2H), 7.35 (d, 1H, J = 4.7), 7.39 (s, 1H), 7.49 (s, 1H), 8.56 (d, 1H , J = 4.7); NMR OCDCF) 99.8,102.2, 106.1,119.9,

123,7, 129,8, 141,2, 147,7, 149,1, 151,4.

b. N- (6,7-Methylenedioxyquinolin-4-yl) butylamine. 4-Chloro-6,7-methylenedioxyquinoline in refluxing phenol (5.5 mol equiv) was stirred for 2.5 h. The temperature was lowered to 100 ° C and butylamine (1.0 mol equiv) was added with stirring. The reaction was then allowed to stir at 100 ° C for several hours and the phenol was removed by Kugelrohr distillation under reduced pressure. The residue was partitioned between CHCl₃ and 10% NaOH. The aqueous phase was repeatedly separated with CHCl₃. All CHCl solutions₃ (initial partition and extracts) were combined and dried (MgSO4) and purified by column chromatography to provide the product; mp. 186-187 ° C; NMR¹H (CD3OD) δ 1.02 (t, 3H, J = 7.2), 1.52 (q, 2H, J = 7.2),

ES 2 311 639 T3

1.75 (q, 2H J = 7.2), 3.33 (q, 2H, J = 7.2), 4.88 (b, 1H), 6.08 (s, 2H), 6.40 (d, 1H, J = 5.6), 7.07 (s, 1H), 7.35 (s, 1H), 8.37 (d, 1H, J = 6.0); NMR¹³C (CH3OD) δ 12.2, 19.3, 29.6, 42.3, 96.9, 97.3, 98.8, 102.3, 112.5,

138.7, 141.5, 147.3, 151.6, 152.8; HRMS calcd for C₁₄H₁₆N₂OR₂: 244.1212; found 244.1222.

c. N- (6,7-Methylenedioxyquinolin-4-yl) -N- (butyl) -2-iodo-4,5-dimethoxybenzamide A 2.0 M solution of oxalyl chloride in CH₂Cl₂ (8.2 mmol) was added to a solution of 2-iodo-5,6-dimethoxybenzoic acid (1.9 mmol) in CH₂Cl₂ anhydrous (~ 60 ml per 10 mmol benzoic acid) and the solution was stirred under reflux for 3 h. The mixture was allowed to cool and then concentrated to dryness in vacuo. To the residue was added a solution of N- (6,7-methylenedioxyquinolin-4-yl) butylamine (400 mg, 1.6 mmol), triethylamine (2 equiv.) In CH₂Cl₂ (~ 60 ml per 4 mmol aminoquinoline). Then, the reaction mixture was stirred under reflux under N₂ for 72 h. The residue was partitioned between CHCl₃ and 10% NaOH. The aqueous phase was repeatedly separated with CHCl₃. All CHCl solutions₃ (initial cast and extracts) were combined and dried (MgSO4). The aqueous phase was neutralized with 20% NaOH and extracted with CHCl3, dried (MgSO4) and evaporated to give the compound: NMR¹H (CDCl₃) δ 0.85 (t, 3H, J = 7.4), 1.20-1.91 (m, 4h), 3.22 (s, 3H), 3.65 (s, 3H), 4, 45 (m, 2H), 6.08 (d, 2H, J = 1.8), 6.31 (s, 1H), 6.97 (s, 1H), 7.17 (d, 1H, J = 4.8), 7.29 (s, 1H), 7.30 (s, 1H), 8.49 (d, 1H, J = 4.8).

Example 10

8,9-Dimethoxy-2,3-methylenedioxy-6H-5-oxa-12-aza-chrysen

A mixture of 4- (2-iodo-4,5-dimethoxybenzyloxy) -6,7-methylenedioxyquinoline (186 mg, 0.4 mmol), Pd (OAc)₂ (18 mg, 0.08 mmol), P (o-tolyl)₃ (49 mg, 0.16 mmol) and silver carbonate (220 mg, 0.8 mmol) was refluxed in DMF (12 ml) and stirred under nitrogen for 25 minutes. The reaction mixture was cooled to room temperature, diluted with chloroform, and filtered through a Celite pad. The filter was washed well with 90:10 chloroformomethanol. Then, the solvent was removed under reduced pressure and the resulting residue was purified by silica gel chromatography using chloroform to give the cyclized compound (30 mg) as a slightly pink solid, in 23% yield; mp. 239-242 ° C (dec.); IR (CHCl₃) 3025, 3009, 29.60, 2928, 2855, 1603, 1526, 1496, 1463, 1346, 1285, 1260, 1243, 1219, 1210, 1181, 1165, 1141, 1041; UV (THF) 4_max= 222, 254, 282, 303, 326 (log ε = 3.98, 4.16, 4.22, 4.12, 4.14); NMR¹H (CDCl3) δ 3.95 (s, 3H), 4.01 (s, 3H), 5.33 (s, 2H), 6.11 (s, 2H), 6.71 (s, 1H), 7.30 (s, 1H), 7.34 (s, 1H), 7.42 (s, 1H), 8.95 (s, 1H); NMR¹³C (CDCl3) δ 56.3, 56.4, 68.9, 97.6, 101.7, 106.0, 108.4, 112, 6, 116, 6, 120, 9, 122, 2, 143 , 4, 147, 2, 147, 8, 149.4, 149, 8, 150, 9, 155.7; HRMS calcd for C₁₉H₁₅DO NOT₅: 337.0950; found: 337.0945.

The intermediate compound 4- (2-iodo-4,5-dimethoxybenzyloxy) -6,7-methylenedioxyquinoline was prepared as follows.

to. Diethyl 3,4-Methylenediogianilinomethylenemalonate. 3,4-methylenedioxyaniline (41.0 g, 0.3 mmol) and diethyl ethoxymethylenemalonate (64.8 g, 0.3 mmol) were refluxed in benzene for 3.5 hours. The solvent was evaporated in vacuo and the residue was washed with petroleum ether to give 88.3 g as a bright grayish brown solid in 96% yield; mp. 99.5-101.0 ° C (Bibl.²²¹ mp. 102 ° C); NMR¹H (CDCl3) δ 1.34 (t, 3H, J = 7.0), 1.40 (t, 3H, J = 7.0) 4.25 (q, 2H, J = 7.0), 4 , 31 (q, 2H, J = 7.0), 6.01 (s, 2H), 6.60 (dd, 1H, J = 8.5, J = 2.2), 6.71 (d, 1H, J = 2.2), 6.81 (d, 1H, J = 8.5), 8.41 (d, 1H, J = 14.0); NMR¹³C (CDCl3) δ 14.4, 14.6, 60.1, 60.4, 92.9, 99.4, 101.8, 108.9, 110.9, 134.3, 145.3, 148 , 9, 152.6, 165.8, 169.3.

b. 4-Hydroxy-6,7-methylenedioxy-3-quinolinecarboxylic acid ethyl ester. Diethyl 3,4-methylenedioxyanilinomethylenemalonate (80.0 g, 0.261 mol) in polyphosphate ester (PPE) (250 g, 0.528 mol) was stirred at 120 ° C with a mechanical stirrer for 2 hours. The reaction mixture was poured into ice water (700 ml) and stirred until homogeneous. Then, the mixture was neutralized (pH 8) with ammonium hydroxide and the precipitate was filtered, washed well with water and dried to give 54.7 g as a brown solid in 80% yield; mp. 277-278 ° C; NMR¹H (DMSO-d₆) δ 1.26 (t, 3H, J = 7.0), 4.16 (q, 2H, J = 7.0), 6.09 (s, 2H), 7.02 (s, 1H), 7.38 (s, 1H), 8.48 (s, 1H).

c. 4-Hydroxy-6,7-methylenedioxy-3-quinolinecarboxylic acid. 4-Hydroxy-6,7-methylenedioxy-3-quinolinecarboxylic acid ethyl ester (45.0 g, 0.172 mol) was added to a solution of KOH (16.8 g, 0.258 mol) in ethanol (500 ml) and the The mixture was refluxed with stirring for 20 hours. Then, the reaction flask was cooled and the ethanol was evaporated under reduced pressure. Then 800 ml of water was added with stirring to completely dissolve the potassium salt and the solution was filtered to remove any impurities. Concentrated HCl was added to bring the mixture to pH 1 and the free acid was removed by filtration and dried in vacuo to give 33.9 g as a beige solid, at 84%; mp. &gt; 300 ° C (Bibl.²²¹ mp. &gt; 290 ° C); NMR¹H (DMSO-d₆) δ 6.27 (s, 2H), 7.30 (s, 1H), 7.55 (s, 1H), 8.72 (s, 1H); NMR¹³C (DMSO-d6) δ 98.5, 101.8, 103.8, 107.9,

120,8, 137,9, 143,5, 148,1, 153,7, 167,4, 177,4.

d. 6,7-Methylenedioxy-4-quinolone. A suspension of 4-hydroxy-6,7-methylenedioxy-3-quinolinecarboxylic acid (30 g, 0.129 mol) in diphenyl ether (320 ml) was heated to reflux with vigorous stirring. The reaction was carefully monitored until clear, approximately 1.5 hr, and then immediately removed from the fire. By then all the starting material had dissolved, but a residue remained

ES 2 311 639 T3 black tarred. The solution was decanted and cooled, allowing the product to precipitate. This material was filtered and washed with ethyl ether to remove all traces of phenyl ether. A second crop was obtained by vigorously washing the tarry residue with ethanol (16 x 250 ml), filtering and evaporating the ethanol, and rinsing the material with ethyl ether. The total yield was 14.9 g as a pale yellow solid at 61%; mp. 285-289 ° C (Bibl.²²¹ mp. 276 ° C); NMR¹H (DMSO-d₆) δ 5.95 (d, 1H, J = 7.3), 6.13 (s, 2H), 6.97 (s, 1H), 7.38 (s, 1H), 7.77 (d, 1H, J = 7.3); NMR¹³C (DMSO-d6) δ 97.5, 102.1, 102.6, 108.7, 119.4, 122.0, 130.8, 138.7, 145.8, 151.7.

and. 4-Chloro-6,7-methylenedioxyquinoline. 6,7-methylenedioxy-4-quinolone (5.0 g, 26.5 mmol) was boiled in POCl3 (75 ml) for 45 min and then cooled. Excess phosphoryl chloride was removed under reduced pressure and ice water (100 ml) was added to hydrolyze any residual phosphoryl chloride. The mixture was basified (pH 9) with ammonium hydroxide and the precipitated solid was filtered. This material was extracted into ethyl ether (8 x 100 ml) and the ether solution was dried (MgSO₄) and evaporated to provide 4.55 g as a white solid at 83%; mp. 127.5-128 ° C (Bibl.²²¹ mp. 129 ° C); NMR¹H (CDCl3) δ 6.15 (s, 2H), 7.35 (d, 1H, J = 4.7), 7.39 (s, 1H), 7.49 (s, 1H), 8.56 (d, 1H, J = 4.7); NMR¹³C (CDCl3) δ 99.8, 102.2, 106.1,

119,9, 123,7, 129,8, 141,2, 147,7, 149,1, 151,4.

F. 4- (2-Iodo-4,5-dimethoxybenzyloxy) -6,7-methylenedioxyquinoline. A mixture of 4-chloro-6,7-methylenedioxyquinoline (414 mg, 2.0 mmol), 2-iodo-4,5-dimethoxybenzyl alcohol (586 mg, 2.0 mmol), and sodium hydride (252 mg of a 60% dispersion, 6.0 mmol) in DMF (25 ml) was stirred at 105 ° C for 90 minutes. The mixture was cooled and a few drops of water were added to quench excess base. The solvent was evaporated under reduced pressure and the residue was purified by chloroform chromatography to give 550 mg as a slightly yellow solid in 59% yield; mp. 196-198 ° C; NMR¹H (DMSO-d6) δ 3.78 (s, 3H), 3.81 (s, 3H), 5.22 (s, 2H), 6.19 (s, 2H), 7.07 (d, 1H , J = 5.4), 7.30 (s, 1H), 7.32 (s, 1H), 7.33 (s, 1H), 7.41 (s, 1H), 8.55 (d, 1H, J = 5.4); NMR¹³C (DMSO-d6) δ 56.5, 56.8, 74.4, 89, 3, 97, 8, 102, 2, 102, 7, 105, 9, 115, 4, 1.16, 2, 117 , 3, 122, 8, 131, 2, 147, 8, 149.8, 149.9, 150.4, 151.2, 160.5; HRMS calcd for C19H ^ OjNl: 465.0073; found: 465.0076.

The 2-iodo-4,5-dimethoxybenzyl alcohol intermediate was prepared as follows.

g. 2-iodo-4,5-dimethoxybenzyl alcohol. To a stirred mixture of 3,4-dimethoxybenzyl alcohol (2.0 g, 11.9 mmol) and silver trifluoroacetate (2.82 g, 12.9 mmol) in anhydrous methylene chloride (50 ml) was added dropwise dropwise a solution of iodine (3.64 g, 14.1 mmol) in methylene chloride (100 ml). After stirring at room temperature for 2 hours, the reaction mixture was filtered and the filtrate was washed with 5% NaHSO₃ (2 x 100 ml) and brine (100 ml), dried (MgSO₄) and the solvent was evaporated in vacuo to provide 3.4 g as a white solid in 97% yield; NMR¹H (CDCl₃) δ 3.90 (s, 3H), 3.91 (s, 3H), 4.61 (s, 2H), 6.95 (s, 1H), 7.04 (s, 1H).

Example 11

Illustrated below are representative pharmaceutical dosage forms containing a compound of the invention ("compound X") for therapeutic or prophylactic use in humans.

<td>(i) Tablet 1</td><td>mg / tablet</td>

<td>'Compound X'</td><td> 100,0</td>

<td>Lactose</td><td> 77,5</td>

<td>Povidone</td><td> 15,0</td>

<td>Croscarmellose sodium</td><td> 12,0</td>

<td>Microcrystalline cellulose</td><td> 92,5</td>

<td>Magnesium stearate</td><td> 3,0</td>

300,0

ES 2 311 639 T3

<td>(ii) Tablet 2</td><td>mg / tablet</td>

<td>'Compound X'</td><td> 20,0</td>

<td>Microcrystalline cellulose</td><td> 410,0</td>

<td>Starch</td><td> 50,0</td>

<td>Sodium starch glycolate</td><td> 15,0</td>

<td>Magnesium stearate</td><td> 5,0</td>

500,0

<td>(Ii) Capsule</td><td>mg / capsule</td>

<td>'Compound X'</td><td> 10,0</td>

<td>Colloidal silicon dioxide</td><td> 1,5</td>

<td>Lactose</td><td> 465,5</td>

<td>Pregelatinized starch</td><td> 120,0</td>

<td>Magnesium stearate</td><td> 3,0</td>

600,0

<td>(iv) Injection 1 (1 mg / ml)</td><td>mg / ml</td>

<td>'Compound X' (free acid form) Dibasic sodium phosphate</td><td> 1,0 12,0</td>

<td>Monobasic sodium phosphate</td><td> 0,7</td>

<td>Sodium chloride</td><td> 4,5</td>

1.0 N sodium hydroxide solution (pH adjustment to 7.0-7.5) qs

Water for injection qs up to 1 ml

<td>(v) Injection 2 (10 mg / ml)</td><td>mg / ml</td>

<td>'Compound X' (free acid form) Monobasic sodium phosphate</td><td> 10,0 0,3</td>

<td>Dibasic sodium phosphate</td><td> 1,1</td>

Polyethylene glycol 400 200.0

01 N sodium hydroxide solution (pH adjustment to 7.0-7.5) qs

Water for injection qs up to 1 ml

ES 2 311 639 T3 (vi) Injection 3 (1 mg / ml) mg / ml 'Compound X' (free base form) 1.0

Citric acid 0.1%

D5W qs up to 1 ml

<td>(vii) Aerosol</td><td>mg / bottle</td>

<td>'Compound X'</td><td> 20,0</td>

<td>Oleic acid</td><td> 10,0</td>

<td>T ricloromonofluoromethane</td><td> 5.000,0</td>

<td>Dichlorodifluoromethane</td><td> 10.000,0</td>

<td>Dichlorotetrafluoroethane</td><td> 5.000,0</td>

The above formulations can be obtained by conventional procedures well known in the pharmaceutical art.

References cited in description

This list of references cited by the applicant is for the convenience of the reader only. It is not part of the European patent document. Although the utmost care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO denies any responsibility in this regard.

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ES 2 311 639 T3 • YAMASHITA et al. Biochemistry, 1991, vol. 30, 5838-5845 [0002] • YAMASHITA et al. Biochemistry, 1992, vol. 31, 12069-12075 [0002] • FEISER; FEISER. Reagents for Organic Synthesis, 1967, vol. 1 [0066] [0125] [0168] [0233] • MARCH, J. Advancad Organic Chemistry. John Wiley &amp; Sons, 1992 [0066] • HOUSE, HO Modern Synthetic Reactions. 1972 [0066] [0168] [0233] • LAROCK, RC Comprehensive Organic Transformations. Wiley-VCH Publishers, 1999 [0066] [0125] [0168] [0233] • MARCH, J. Advanced Organic Chemistry. John Wiley &amp; Sons, 1992 [0125] [0168] [0233] • HOUSE, HO Modern Synthetic Reactions. 1972 [0125] • GREENE, TW; WUTZ, PGM Protecting Groups In Organic Synthesis. John Wiley &amp; Sons, Inc, 1991 [0237] • MAKHEY, D. et al. Bioorg. Med. Chem., 2000, vol. 8, 1-1 [0259] • MANIATIS, T. et al. J. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory, 149-185 [0259] • MANIATIS, T .; FRITSCH, EF; SAMBROOK, J. Molecular Cloning, a Laboratory Manual. Cold Spring Harbor Laboratory, 1982, 149-185 [0259] • LIU, LF; ROWE, TC; YANG, L .; TEWEY, KM; CHEN, GLJ Biol. Chem., 1983, vol. 258, 15365 [0259] • B. GATTO et al. Cancer Res., 1996, vol. 56, 2795-2800 [0259] • DR. LF LIU. Department of Pharmacology, The University of Medicine and Dentistry of New Jersey. Robert Wood Johnson Medical School [0259] [0262] • CHEN AY et al. Cancer Res., 1993, vol. 53, 1332 [0262] • MOSMANN, TJJ Immunol. Methods, 1983, vol. 65, 55 [0262] • CARMICHAEL, J. et al. Cancer Res., 1987, vol. 47, 936 [0262] • DR. TOSHIWO ANDOH. Anchi Cancer Research Institute [0262] • ANDOH, T .; OKADA, K. Adv. in Pharmacology, 1994, vol. 29B, 93 [0262] • RUBIN et al. J. Biol. Chem., 1994, vol. 269, 2433-2439 [0262]