HK1024694B - Pro-drugs and counterparts of camptothecin, their application as medicines - Google Patents

Description

Prodrug forms and analogs of camptothecin, and pharmaceutical uses thereof

Camptothecin is a natural compound that was first isolated from leaves and bark of the chinese plant camptotheca acuminata (see Wall et al, j. amer. chem. soc. 88: 3888 (1966)). The camptothecin is prepared by indolizino [1, 2-b ]]Five-ring compounds fused from quinoline fragments (rings A, B, C and D) and α -hydroxy lactones having six-membered rings (ring E). The carbon at position 20 with the alpha-hydroxyl group is asymmetric and imparts optical rotatory power to the molecule. The natural form of camptothecin has the absolute "S" configuration at the 20-carbon and is represented by the following formula:

camptothecin and its analogs have antiproliferative activity against several cancer cell lines, including human tumor cell lines of the colon, lung and breast (Suffness, M et al, alkaloid chemistry and pharmacology, Bross A et al, Vol.25, page 73 (academic Press, 1985)). This indicates that the antiproliferative activity of camptothecin is related to its inhibitory activity on DNA topoisomerase I.

In addition, camptothecin and certain analogs thereof are insoluble in water, which makes their administration by parenteral route difficult. Water-soluble derivatives of camptothecin have been prepared, which carry salt-forming substituents in the a and B rings (see, e.g., US4,981,968, US5,049,668, EP540,099). However, these products show a lower antitumor activity than the water-insoluble derivatives. Other water-soluble derivatives of camptothecin have also been prepared in which the hydroxyl group at the 20-position is esterified with an acid bearing a salifiable group such as glycine (see, US 4943579 and PCT WO 96/02546). These derivatives are referred to by those skilled in the art as "prodrug forms" because they are not biologically active per se, but only after first metabolism when administered to a patient. The pro-drug form of the alpha-hydroxy lactone analogue of camptothecin has been shown to have good anti-tumor effects in animals and clinically, but is accompanied by destructive side effects, such as severe diarrhea, which are dangerous to the life of the patient. Thus, there is a need to develop more effective and better tolerated water-soluble analogues of camptothecin.

Furthermore, it has been established that alpha-hydroxy lactones are essential for both in vivo and in vitro camptothecin activity (camptothecin: a novel anti-cancer agent, published by Putmesil, M et al, page 27 (CRC Press, 1995); Wall M et al, cancer research 55: 753 (1995); Hertzberg et al, J. Pharmacochemistry 32: 715(1982) and Crow et al, J. Pharmacochemistry 35: 4160 (1992)). However, the applicants have found that beta-hydroxy lactones having a 7-membered ring have biological activity comparable to or greater than alpha-hydroxy lactones (unpublished PCT application FR 96/00980). The present invention relates to novel derivatives of camptothecin analogs, wherein a 7-membered β -hydroxy lactone replaces the natural α -hydroxy lactone of camptothecin. Beta-hydroxy lactone refers to a lactone having another carbon atom between the carboxyl carbon and the hydroxyl-bearing alpha-carbon in the alpha-hydroxy lactone.

Two approaches have been chosen to increase the water solubility of this camptothecin analogue: the first grafts oxazines to the a-ring of the molecule, while the second designs the prodrug form by acylating the hydroxyl functionality of the β -hydroxy lactone.

In particular, among such novel camptothecin analogs, the compounds of the present invention are either analogs modified by the addition of an oxazine ring at carbons 10 and 11, or are in the form of prodrugs wherein a β -hydroxy lactone replaces the natural α -hydroxy lactone of camptothecin. Thus, the compounds of the present invention are camptothecin β -hydroxy lactone analogs or water-soluble prodrugs having an oxazine ring grafted thereto and have unexpectedly strong biological activity in the art.

More specifically, the subject of the present invention is the compounds of formula (I) and formula (II), their racemates, enantiomeric forms or any mixture of these forms or a pharmaceutically acceptable salt thereof:

R₁represents lower alkyl, lower alkenyl, lower alkynyl, lower haloalkyl, lower alkoxy-lower alkyl or lower alkylthio-lower alkyl;

R₂、R₃and R₄Independently represent H, halogen, lower haloalkyl, lower alkyl, lower alkenyl, cyano, lower cyanoalkyl, nitro, lower nitroalkyl, acylamino, lower acylaminoalkyl, hydrazino, lower hydrazinoalkyl, azido, lower azidoalkyl, (CH)₂)_mNR₆R₇、(CH₂)_mOR₆、(CH₂)_mSR₆、(CH₂)_mCO₂R₆、(CH₂)_mNR₆C(O)R₈、(CH₂)_mC(O)R₈、(CH₂)_mOC(O)R₈、O(CH₂)_mNR₆R₇、OC(O)NR₆R₇、OC(O)(CH₂)_mCO₂R₆Or (CH)₂)_n[N＝X]、OC(O)[N＝X]、(CH₂)_mOC(O)[N＝X](in the present invention, [ N ═ X]Represents a 4-to 7-membered heterocyclic group containing a nitrogen atom N, N being one atom of the heterocyclic group, X representing the remaining atoms necessary to constitute the heterocyclic ring, selected from O, S, CH₂、CH、N、NR₉And COR₁₀) Unsubstituted or substituted (i.e. one to four times substituted on the aryl or heterocycle), aryl or lower arylalkyl, wherein the substituents are lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkyl, lower alkoxy or lower alkoxy lower alkyl, or R₂And R₃Or R₃And R₄Together form a 3-or 4-membered chain, wherein the elements of the chain are selected from CH, CH₂O, S, N or NR₉；

R₅Represents H, halogen, lower haloalkyl, lower alkyl, lower alkoxy-lower alkyl, lower alkylthio-lower alkyl, cycloalkyl-lower alkyl, cyano, cyanoalkyl, lower alkyl-lower sulfonylalkyl, lower hydroxyalkyl, nitro, (CH)₂)_mC(O)R₈、(CH₂)_mNR₆C(O)R₈、(CH₂)_mNR₆R₇、(CH₂)_mN(CH₃)(CH₂)_nNR₆R₇、(CH₂)_mOC(O)R₈、(CH₂)_mOC(O)NR₆R₇、(CH₂)_mS(O)_qR₁₁、(CH₂)_mP(O)R₁₂R₁₃、(CH₂)₂P(S)R₁₂R₁₃Or (CH)₂)_n[N＝X]、OC(O)[N＝X]、(CH₂)_mOC(O)[N＝X]ASubstituted or substituted (i.e., one to four times substituted on aryl or heteroaryl) aryl or lower arylalkyl, wherein the substituents are lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkyl, lower alkoxy, or lower alkoxy lower alkyl;

R₆and R₇Independently represent H, lower alkyl, lower hydroxyalkyl, lower alkyl lower aminoalkyl, cycloalkyl lower alkyl, lower alkenyl, lower alkoxy lower alkyl, lower haloalkyl or substituted (i.e. substituted one to four times on the aryl) or unsubstituted aryl or lower arylalkyl, wherein the substituents are lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkyl, lower alkoxy or lower alkoxy lower alkyl;

R₈represents H, lower alkyl, lower hydroxyalkyl, amino, lower alkylamino, lower alkyllower aminoalkyl, lower aminoalkyl, cycloalkyl lower alkyl, lower alkenyl, lower alkoxy, lower alkoxylower alkyl, lower haloalkyl or substituted (i.e. substituted one to four times on aryl) or unsubstituted aryl or lower arylalkyl, wherein the substituents are lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkyl, lower alkoxy or lower alkoxylower alkyl;

R₉represents H, lower alkyl, lower haloalkyl, aryl or aryl substituted by one or more of the following: lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkyl, lower alkoxy or lower alkoxy lower alkyl;

R₁₀represents H, lower alkyl, lower haloalkyl, lower alkoxy, aryl or aryl substituted by one or more groups (i.e. having one to four substituents on the aryl), the substituents being selected from lower alkyl, lower haloalkyl, lower hydroxyalkyl or lower alkoxyA lower alkyl group;

R₁₁represents a lower alkyl group, an aryl group, or (CH)₂)_mOR₁₄、(CH₂)_mSR₁₄、(CH₂)₂NR₁₄R₁₅Or (CH)₂)_m[N＝X]；

R₁₂And R₁₃Independently represent lower alkyl, aryl, lower alkoxy, aryloxy or amino;

R₁₄and R₁₅Independently represents H, lower alkyl or aryl;

R₁₆represents H OR OR₂₁；

R₁₇Is represented by OR₆Or NR₆R₇；

R₁₈And R₁₉Independently represents H, halogen, lower alkyl, lower alkoxy or hydroxy;

R₂₀represents H or halogen;

R₂₁represents H, lower alkyl, CHO or C (O) (CH)₂)_mCH₃；

R_pRepresents H or a group which is easily cleavable, preferably corresponding to the formula-C (O) -A-NR₂₂R₂₃Wherein A represents a linear or branched alkylene group, which alkylene group is optionally substituted by a group selected from free, esterified or salified hydroxyl, halogen, free, esterified or salified carboxyl, amino, mono-or dialkylamino and R₂₂And R₂₃Independently represent H, lower alkyl, lower hydroxyalkyl, lower alkyl lower aminoalkyl, cycloalkyl lower alkyl, lower alkenyl, lower alkoxy lower alkyl, lower haloalkyl or unsubstituted or substituted (i.e. having one to four substituents on the aryl) aryl or lower arylalkyl, the substituents being selected from lower alkyl, halogen, nitro, amino, lower alkylamino, lower haloalkyl, lower hydroxyalkylLower alkoxy or lower alkoxy lower alkyl;

m is an integer between 0 and 6;

n is 1 or 2; while

q represents an integer of 0 to 2; and is

[N＝X]Represents a 4-to 7-membered heterocyclic group, X represents the chain required to complete said heterocyclic group and is selected from O, S, CH₂、CH、N、NR₉And COR₁₀；

It should be understood that when R is_pWhen it is a hydrogen atom, R₃And R₄Together form a 3 or 4 membered chain.

Herein, the term "lower" in connection with alkyl, alkylthio and alkoxy means a straight or branched chain saturated aliphatic hydrocarbon group having 1 to 6 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, methylthio, ethylthio, methoxy and ethoxy. The term "lower" in connection with the term alkenyl or alkynyl refers to a group containing 2 to 6 carbon atoms and one or more double or triple bonds, for example, vinyl, allyl, isopropenyl, pentenyl, hexenyl, propenyl, ethynyl, propynyl and butynyl. The term cycloalkyl refers to a ring containing 3 to 7 carbons, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. The term aryl refers to mono-, di-or tricyclic hydrocarbon compounds in which at least one ring is aromatic and each ring contains up to 7 carbon atoms, e.g. phenyl, naphthyl, anthracenyl, biphenyl or indenyl. The term halogen refers to chlorine, bromine, iodine or fluorine. Groups corresponding to the terms lower haloalkyl, lower cyanoalkyl, lower nitroalkyl, lower amidoalkyl, lower hydrazinoalkyl, lower azidoalkyl, lower arylalkyl, lower hydroxyalkyl, lower alkoxy lower alkyl, lower alkylthio lower alkyl and lower alkyl lower sulfonylalkyl are substituted with one to three halogen, cyano, nitro, amido, hydrazino, azido, aryl, hydroxy, lower alkyl, lower alkylthio or lower sulfonylalkyl groups, respectively. Lower alkylamino may contain one or two lower alkyl groups, e.g. as shown in TableShown as NHCH₃、NHCH₂CH₃、N(CH₃)₂Or N (CH)₃)(CH₂CH₃). The term free, esterified, etherified or salified hydroxy refers to OH, OCOR₂₆、OR₂₇Groups and alkoxides thereof.

The compounds of the present invention have two possible enantiomeric forms, namely the "R" and "S" configurations. The present invention includes both enantiomeric forms and mixtures thereof, including "RS" racemic mixtures. For the sake of simplicity, when no particular configuration in a structural formula is indicated, it is understood to mean both enantiomers and mixtures thereof.

For the prodrug forms of the invention (wherein R is_pNot a hydrogen atom), preferably a product of the formula I.

Examples of substituted camptothecins useful as starting materials are found in U.S. Pat. Nos. 4473692, 4604463, 4894956, 5162532, 5395939, 5315007, 5264579, 5258516, 5254690, 5212317 and 5341745, PCT patent applications US91/08028, US94/06451, US90/05172, US92/04611, US93/10987, US91/09598, EP94/03058 and EP95/00393 and European patent applications 325247, 495432, 321122 and 540099.

Such compounds containing oxazine rings:

-reacting a compound of beta-hydroxy lactone of formula D:wherein R is₃Is hydroxy, R₄Is H and R₁、R₂、R₅、R₁₈、R₁₉And R₂₀As defined above, with a primary amine under Mannich reaction conditions to give a β -hydroxy lactone compound of formula Ia

Wherein R is₁、R₂、R₅、R₉、R₁₈、R₁₉And R₂₀As defined above.

The process comprises heating the starting material in the presence of a primary amine such as benzylamine, formaldehyde in an acid reagent such as acetic acid or propionic acid at a temperature of from 30 to 80 ℃ for from 0.5 to 5 hours. Alternatively, the acetic acid suspension of the starting material is heated with tri-N-substituted hexahydrotriazines, such as hexahydro-1, 3, 5-trimethyltriazine, 1, 3, 5-triethylhexahydrotriazine or 1, 3, 5-tribenzylhexahydrotriazine, at 30 to 80 ℃ for 0.5 to 5 hours.

The lactone of the formula Ia is optionally ring-opened in basic medium, in order to prepare the compound of the formula IIa after neutralization

Wherein R is₁、R₂、R₅、R₉、R₁₇、R₁₈、R₁₉And R₂₀As defined above; r₁₆Is represented by OR₂₁Wherein R is₂₁Represents H or lower alkyl; and R is₁₇Is represented by OR₆Or NHR₆And R is₆Represents H, lower alkyl, cycloalkyl, lower alkylcycloalkyl, lower alkenyl, lower alkyl lower alkoxy or aryl or lower alkylaryl.

The compound of the formula D or Ia is optionally acylated, preferably with C (O) -A-N-R as defined above₂₂R₂₃Acylation of derivatives of the group to prepare beta-hydroxy lactone compounds of the general formula Ib, i.e. R_pFormula I (prodrug form of the invention) other than H.

The hydroxy acid IIb can be prepared by ring opening lactone Ib in the same manner as ring opening lactone Ia.

In the above-mentioned methods, standard protection methods (Greene.T.,. protective groups for organic Synthesis) 10-86(John Wiley) can be followed if necessary&Sons 1981)) to protect R₂、R₃、R₄And R₅A group. If at least R₂₂And R₂₃Where one of the groups is H, or contains at least one functional group such as a primary or secondary amine that is chemically incompatible with the acylation process, it may be desirable to use a protecting group that is not affected by the acylation conditions. Are commonly used to protect aminesThe protecting group of (2) is tert-Butyloxycarbonyl (BOC). The acylation reaction is then carried out as described above and the protecting group is cleaved, for example by treatment with trifluoroacetic acid for BOC, to produce the compound of formula (I) or (II). The use of protecting groups is known to the person skilled in the art (see, for example, Greene.T., "protecting groups for organic Synthesis" John Wiley&Sons 1981)。

The compound of formula D is prepared as follows:

compounds of the formula M

Wherein R is₁、R₁₈And R₁₉As defined above and R₂₀Represents H or a halogen atom, with a 2-halo-3-quinoline-methanol of the general formula N,wherein R is₂、R₃、R₄And R₅As defined above and X represents a halogen atom, to prepare a compound of formula O

Wherein R is₁、R₂、R₃、R₄、R₅、R₁₈、R₁₉、R₂₀And X is as defined above;

-then cyclizing the compound of formula O to give the compound of formula D as defined above.

In the above-mentioned methods, standard protection methods (Greene.T., "protecting groups for organic Synthesis" 10-86(John Wiley) can be followed, if necessary&Sons 1981)) to protect R₁、R₂、R₃And R₄A group. The reaction to form the compound O starting from the compounds of the formulae M and N can be carried out according to methods known to the person skilled in the art, the so-called Mitsunobu reaction (see Mitsunobu, O. et al, Synthesis, p.1 (1981)). By using phosphines such as triphenylphosphine and azodicarboxylate derivatives such as diethyl azodicarboxylate in an aprotic solvent such as tetrahydrofuranOr N, N-dimethylformamide to replace the hydroxyl function of compound N with a nucleophile such as compound M or its deprotonated derivative. The cyclization of compound O is preferably carried out in the presence of a palladium catalyst, for example palladium diacetate, under basic conditions (for example alkali metal acetates optionally mixed with a phase transfer agent such as tetrabutylammonium bromide), in an aprotic solvent such as acetonitrile or N, N-dimethylformamide at temperatures of from 50 to 120 ℃ (R.Grigg et al, tetrahedron, p. 4003 (1990)).

The compounds of formula M can be prepared according to the following methods characterized by:

-of the formula wherein R₁And R₂₀As defined above and R₂₄The carbonyl group of pyridine representing halogen atom or lower alkoxy group is protected with acetal functional group,thereby preparing a compound of formula F

Wherein R is₁、R₂₀And R₂₄The above definitions and the Z and Z' groups independently represent lower alkyl or together form a 2 to 4 membered saturated hydrocarbon:

-introduction of a hydroxymethyl function into a compound of formula F to prepare a compound of formula G

Wherein R is₁、R₂₀、R₂₄Z and Z' are as defined above,

subsequent protection of the alcohol function of the compound of formula G to prepare a compound of formula H

Wherein R is₁、R₂₀、R₂₄Z and Z' are as defined above and R₂₅Represents a protecting group for an alcohol functional group.

Will go throughDeprotection of the acetal of a compound of formula H to produce a compound of formula I

Wherein R is₁、R₂₀、R₂₄And R₂₅The definition is as above-mentioned,

treatment of a compound of formula I' with a functionalised alkylating agent to prepare a compound of formula J

Wherein R is₁、R₂₀、R₂₄And R₂₅As defined above, R₁₇、R₁₈And R₁₉The definition is shown in a general formula II,

-protecting group R of compound of formula J₂₅Cleavage to prepare compounds of the formula K

Wherein R is₁、R₁₈、R₁₉、R₂₀And R₂₄As defined above, and R₁₇Is represented by OR₆Or NHR₆，R₆Represents H, lower alkyl, cycloalkyl, lower alkylcycloalkyl, lower alkenyl, lower alkyl-lower alkoxy or aryl or lower alkylaryl,

cyclisation of a Compound of the formula K to a Compound of the formula L

Wherein R is₁、R₁₈、R₁₉、R₂₀And R₂₄As defined above, and finally

R of the Compound L₂₄Conversion of the radical into a carbonyl group to give the compound of the formula M

Wherein R is₁、R₁₈、R₁₉And R₂₀Is defined asThe above.

The carbonyl function of 4-acyl-2-halopyridines (obtained, for example, according to Lammantina J.L., J.Heterocycl. chem. 20, p.553 (1983)) is preferably protected by an acetal function, preferably a cyclic acetal, according to methods known to the person skilled in the art (Greene.T.; protective groups for organic Synthesis 10-86(John Wiley. RTM.) (see U.S.A.)&Sons 1981)). The intermediate prepared as above is treated with an alcoholate of sodium or potassium in an aprotic solvent (e.g. acetonitrile) or the alcohol from which the alcoholate is prepared, at a temperature of 0 ℃ to 100 ℃, to give the compound of general formula F. Its 3-position is lithiated by treatment with an aryl or alkyl lithium (e.g., * -based lithium) in an ether solvent such as tetrahydrofuran at-100 to 0 ℃. A formylating electrophile such as N, N-dimethylformamide is added to the lithiated intermediate so obtained, and after hydrolysis, the aldehyde so obtained is treated with a reducing agent such as sodium borohydride to produce a compound of formula G. The protection of the alcohol function of compound G is carried out according to standard conditions known to the person skilled in the art, thus preparing the compound of general formula H. Examples of protecting groups for alcohol functional groups include those that form ethers (i.e., methyl, methoxymethyl, tetrahydropyranyl, 2-methoxyethoxymethyl, benzyloxymethyl, t-butyl, and substituted or unsubstituted benzyl), and those that form esters (formate, acetate, and isobutyrate). Other examples of protecting groups for primary hydroxyl groups are described in Greene.T., "protecting groups for organic Synthesis" 10-86(John Wiley&Sons 1981). Deprotection of a compound of formula H to produce a compound of formula I' is carried out by maintaining R₂₅The integrity of the groups is carried out under selected conditions, for example, by treatment under acidic conditions (e.g., with trifluoroacetic acid). The conditions chosen for the protection and deprotection of functional groups are known to those skilled in the art (Greene.T.,. protective groups for organic Synthesis 10-86(John Wiley)&Sons 1981)). The reaction of treating compound I' with a functionalized alkylating agent to prepare the beta-hydroxy ester of formula J can be carried out using a lithium enolate or a zinc derivative of a carboxylic acid ester in an anhydrous aprotic solvent such as tetrahydrofuran. Protecting group R of compound of formula J₂₅Cleaved under deprotection conditions known to those skilled in the art to producePreparing the compound of the general formula K. For example, when R is₂₅When it is benzyl, the alcoholic solution of the compound of the formula J and the palladium catalyst added thereto are placed in a hydrogen atmosphere at a pressure of 0.5 to 10 bar. The ring closure reaction of the compound of formula K thus obtained can be carried out under acidic conditions (e.g., by treatment with trifluoroacetic acid or hydrochloric acid gas dissolved in an anhydrous solvent such as dichloromethane or dioxane) to prepare a β -hydroxy lactone ring which is a 7-membered ring, such as a compound of formula L. The compounds of the formula L can be converted into pyridones of the formula M, for example, by treatment with hot perchloric acid or by treatment with trimethylsilyl iodide.

The 2-halo-3-quinolinemethanol of the general formula N can be prepared starting from the acetanilide of the general formula P,

wherein R is₂、R₃And R₄The definitions are as described for the compounds of the general formulae I and II. In the following methods, standard protection methods (Greene.T.,. protective groups for organic Synthesis) 10-86(John Wiley) can be followed, if desired&Sons 1981)) to protect R₂、R₃And R₄A group.

The compound of formula N is then obtained according to the following process: the aniline of said formula P is N-acetylated by treatment with an acylating agent such as acetic anhydride. The N-acetylaniline so produced is treated at 50 to 100 deg.C, preferably 75 deg.C, with a so-called Vilsmeyer reagent known to those skilled in the art (prepared from phosphoryloxy chloride acting on N, N-dimethylformamide at 0 to 10 deg.C) to produce the corresponding 2-chloro-3-quinolinecarboxaldehyde (see, for example, Meth-Cohn et al, J.Chem.Soc., Perkin Trans.I, p.1520 (1981); Meth-Cohn et al, J.Chem.Soc., Perkin Trans.I, p.2509 (1981) and Nakasimhan et al, J.Chem.Soc., 112, p.4431 (1990)). The chlorine at the 2-position of the 2-chloro-3-quinolinecarboxaldehyde may be replaced by iodine or bromine by heating the product in an inert solvent such as acetonitrile in the presence of an iodine or bromine salt (e.g., sodium iodide or tetrabutylammonium bromide). Trace amounts of acid such as concentrated hydrochloric acid may be required to catalyze this conversion process. The 2-halo-3-quinolinecarboxaldehyde is readily reduced to the corresponding 2-halo-3-quinolinemethanol of general formula N, under standard conditions known to those skilled in the art, for example by treatment with sodium borohydride in an alcoholic solvent (e.g.methanol) at a temperature of from 0 to 40 ℃.

The compounds of formula N may also be prepared according to the following method: anilines of the general formula P as defined above are acylated by reaction with a nitrile, such as chloroacetonitrile or propionitrile, in the presence of boron trichloride and another Lewis acid, such as aluminum trichloride, titanium tetrachloride or diethylaluminum chloride, in an aprotic solvent or a mixture of aprotic solvents, followed by hydrolysis (cf. Sugasawa T. et al, journal of the American society for chemistry, 100, p.4842 (1978)). The intermediate prepared above is then treated with ethylmalonyl chloride in an aprotic solvent such as acetonitrile in the presence of a base such as triethylamine, followed by treatment with an alkali metal alcoholate such as sodium methylate in methanol to prepare the 4-substituted ethyl 2-hydroxy-3-quinolinecarboxylate. It was converted to ethyl 2-chloro-3-quinolinecarboxylate by treatment with phosphoryloxy chloride. When quinoline carries a chloromethyl group in the 4-position, nucleophilic substitution can be carried out by treatment with secondary amines such as dimethylamine, N-methylpiperazine, morpholine or piperidine. This ethyl 2-chloro-3-quinolinecarboxylate is then reduced with diisobutylaluminum hydride in an aprotic solvent such as dichloromethane to give the 2-chloro-3-quinolinecarboxyl alcohol of formula N. Analogues of intermediate compound (N) have been described in the literature, in particular in PCT application 95/05427.

Certain compounds of the present invention may be prepared in the form of pharmaceutically acceptable salts according to conventional methods. For example, acceptable salts include, but are not limited to, addition salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, and nitric acid, or organic acids such as acetic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, citric acid, lactic acid, methanesulfonic acid, p-toluenesulfonic acid, 1' -methylene-bis (2-hydroxy-3-naphthoic acid), salicylic acid, oxalic acid, and stearic acid. Salts with bases such as sodium hydroxide or potassium hydroxide are also part of the invention when they are available. Other examples of pharmaceutically acceptable salts are described in "pharmaceutically acceptable salts", journal of pharmacology 66: 1(1977).

The compounds of the invention have beneficial pharmaceutical properties. The compounds of the invention therefore have an inhibitory effect on topoisomerase I and/or II and have antitumor activity. The prior art suggests that the compounds of the present invention have antiparasitic and/or antiviral activity. The compounds of the invention can thus be used in different therapeutic applications.

The pharmaceutical properties of the compounds of the invention are illustrated in the experimental section below.

By administering to a patient a therapeutically effective amount of a compound of formula (I) or (II), the compound inhibits topoisomerase, e.g. type I and/or II, in a patient, e.g. a mammal such as a human.

The compounds of the invention also have antitumor activity. They are useful for treating tumors, such as tumors that express topoisomerase, by administering to a patient a therapeutically effective amount of a compound of formula (I) or (II). Examples of tumors or cancers include cancers that occur in the esophagus, stomach, intestine, rectum, oral cavity, pharynx, larynx, lung, colon, breast, uterus, endometrium, ovary, prostate, testis, bladder, kidney, liver, pancreas, bone, connective tissue, skin, eye, brain and central nervous system, as well as thyroid cancer, leukemia, hodgkin's disease, lymphomas other than those associated with hodgkin's disease, multiple myeloma, and the like.

They are useful in the treatment of parasitic infections, as well as viral infections and diseases, by inhibiting blood flagellates (e.g., in trypanosomes or leishmania infections) or by inhibiting plasmodium (e.g., in malaria).

These properties make the products of formula (I) or (II) suitable for pharmaceutical applications. The subject of the invention is also the products of formula (I) or (II) above and the addition salts of said products of formula (I) or (II) with pharmaceutically acceptable inorganic or organic acids as medicaments, and also the pharmaceutical compositions containing at least one of the medicaments as defined above as active ingredient.

Accordingly, the present invention relates to pharmaceutical compositions containing a compound of the present invention or an addition salt thereof with a pharmaceutically acceptable acid in admixture with a pharmaceutically acceptable carrier selected according to the method of administration, e.g., oral, intravenous, intraperitoneal, intramuscular, transdermal or subcutaneous. The pharmaceutical composition (e.g., therapy) may be a solid, a liquid, a liposome, or a lipid micelle.

The pharmaceutical composition may be in solid form, for example, powders, pills, granules, tablets, liposomes, gelatin capsules or suppositories. The pill, tablet or gelatin capsule may be coated with a substance which protects the composition in the stomach from the action of gastric acid or enzymes for a sufficient period of time to allow the composition to pass undigested into the small intestine. The compounds may also be administered topically, for example at the site of tumor pathogenesis. The compounds may also be administered in a sustained release manner (e.g., a sustained release composition or infusion pump). Suitable solid carriers are, for example, calcium phosphate, magnesium stearate, magnesium carbonate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and waxes. The pharmaceutical compositions containing the compounds of the invention may also be in liquid form, e.g., solutions, emulsions, suspensions or sustained release formulations. Suitable liquid carriers may be, for example, water, organic solvents such as glycerol or glycols such as polyethylene glycol, and mixtures thereof with water in varying proportions.

The subject of the invention is also the use of a product of formula (I) or (II) as defined above for the preparation of a medicament: drugs that inhibit topoisomerase and in particular type I or II topoisomerase, drugs for treating tumours, drugs for treating parasitic infections and drugs for treating viral infections.

The dosage of the compounds of the present invention for use in the treatment of the above-mentioned diseases or disorders will vary depending on the method of administration, the age and weight of the patient and the physical condition of the patient, and will be ultimately at the discretion of the attendant physician or veterinarian. The amount determined by the attending physician or veterinarian is referred to herein as a "therapeutically effective amount".

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Also, all publications, patent applications, all patents, and all other references mentioned herein are incorporated by reference.

The following examples are intended to illustrate the above process and should not be construed as limiting the scope of the invention in any way.

Experimental part

Preparation example 1: 5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

1, a.4-ethyl-3, 4-dihydroxy-1H-pyrano [3 ', 4': 6, 7 indolizino [1, 2-b ] quinolin-14 (4H, 12H) -one

Sodium borohydride (14g, 370mmol) was added portionwise to a suspension of (S) - (+) -camptothecin (14g, 40mmol, available from different places such as Aldrich Chemical Co. (Milwaukee, WI)) in methanol (750ml) and the resulting mixture was gently heated to 55 ℃ to give a clear solution which was stirred at room temperature for 16 hours. Then, the solvent was evaporated under reduced pressure, and the residue was recovered in water (250ml), neutralized by adding acetic acid (21ml), and left at 4 ℃ for 2 hours. The resulting suspension was filtered and washed successively with cold water, acetone and ether, and the resulting product was dried under reduced pressure to form the desired compound as a white solid, m.p.280 ℃.

1, b.8-Formyloxymethyl-7-propionylindolizino [1, 2-b ] quinolin-9 (11H) -one

A solution of sodium metaperiodate (14g, 65mmol) in water (140ml) was added dropwise to a solution of 4-ethyl-4, 5-dihydroxy-1H-pyrano [3 ', 4': 6, 7] indolizino [1, 2-b ] quinolin-14 (4H, 12H) -one (13.4g, 38mmol) in glacial acetic acid (720ml) and the resulting solution is stirred at room temperature for 1 hour. The reaction mixture was then poured into an ice/water mixture (650ml) and the resulting suspension was stirred for a further half an hour, then filtered and washed successively with water, isopropanol and diethyl ether to give, after drying under reduced pressure, the product (11.5g) as a pale yellow solid, m.p. > 200 ℃ (decomposed).

beta-Ethyl-beta-hydroxy-beta- (8-hydroxymethyl-9-oxo (11H) -indolizino [1, 2-b ] quinolin-7-yl) propionic acid tert-butyl ester

A suspension of zinc (6.5g, 100mmol) stirred with a magnetic stirrer under argon atmosphere in dry ether (50ml) was activated by dropwise addition of chlorotrimethylsilane (0.75ml, 5.7 mmol). Stirring is continued for 15 minutes at room temperature and the reaction medium is heated to reflux. The hot bath was removed and t-butyl bromoacetate (15ml, 100mmol) was added dropwise at a rate that ensured continued reflux. It was placed in an external heat source again and heating was continued for 1 hour. The ether solution obtained from the Reformatsky reagent was left to cool to room temperature and then transferred to 8-formyloxymethyl-7-propionylindolizino [1, 2-b ] with a catheter under argon]Quinolin-9 (11H) -one (1.6g, 4.7mmol) in dry tetrahydrofuran (40 ml). The reaction mixture was stirred at reflux for 1 hour, then cooled to room temperature and quenched by the addition of saturated ammonium chloride (100ml) and extracted with chloroform (3X 100 ml). The combined chloroform extracts were dried over sodium sulfate, evaporated and chromatographed on a silica gel column (1-2% methanol/dichloromethane) to give 0.64g of the product (31%) as a pale yellow solid, m.p.146-149 ℃. NMR-1H (CDCl)₃)：0.93(t，3H)；1.37(s，9H)；1.99(m，2H)；2.97(dd，2H)；3.5(se，1H)；5.10(s，2H)；5.24(s，2H)；7.40(s，1H)；7.59(t，1H)；7.83(t，1H)；7.90(d，1H)；8.20(d，1H)；8.34(s，2H).NMR-C13(CDCl₃)：8.18；27.90；34.59；45.34；49.91；58.55；77.39；82.42；100.52；127.67；127.97；128.10；128.64；129.44；129.79；130.42；130.99；142.86；148.69；152.75；155.16；162.38；172.24.IR(KBr)：764；1016；1157；1580；1651；1726.

1, d.5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

Tert-butyl β -ethyl- β -hydroxy- β - (8-hydroxymethyl-9-oxo (11H) -indolizino [1, 2-b ] quinolin-7-yl) propanoate (1.45g, 3.32mmol) was dissolved in anhydrous dichloromethane (25ml) and treated with a saturated solution of hydrogen chloride in dichloromethane (100 ml). The resulting mixture was maintained at-20 ℃ for 16 hours. The precipitate was filtered off, washed with methanol and dried under reduced pressure to yield 662mg (55%) of the title compound as a yellow solid, m.p. > 300 ℃. NMR-1H (DMSO): 0.90(t, 3H); 1.20(q, 2H); 3.27(dd, 2H); 5.29(s, 2H); 5.49(dd, 2H); 7.42(s, 1H); 7.73(t, 1H); 7.90(t, 1H); 8.16(t, 2H); 8.71(s, 1H). NMR-C13 (DMSO): 8.45 of; 36.48, respectively; 42.54, respectively; 50.68; 61.44; 73.34; 99.78; 122.71, respectively; 127.83, respectively; 128.15 of the total weight of the powder; 128.75; 129.08, respectively; 130.07, respectively; 130.61, respectively; 131.81, respectively; 144.66, respectively; 148.04, respectively; 152.80, respectively; 155.91, respectively; 159.26, respectively; 172.08.IR (KBr): 761; 1127, and (b); 1204; 1285, mixing the raw materials; 1580; 1653; 1757.

preparation example 2: 5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': resolution of 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

A mixture of β -ethyl- β -hydroxy- β - (8-hydroxymethyl indolizino [1, 2-b ] quinolin-9- (11H) -one-7-yl) propanoic acid (19.5g, 51mmol) and L- (-) - α -methylbenzylamine (12.12g, 100mmol) in pure ethanol (11) was heated to boiling, filtered while heating and allowed to stand for 68 hours. The precipitate was filtered off and washed with ethanol and diethyl ether to give 9.8g of a white solid. Two peaks were analyzed by high pressure liquid chromatography on Chiral stationary phase (100X 4mM "Chiral HPLC on Chiral-AGP column (Chromatch, Stockholm, Sweden), eluent 2% acetonitrile in 10mM phosphate buffer pH6.9, eluting peaks at 4.5 and 7.5 min) with integrated areas accounting for 24% and 76% of the total area of the two peaks, respectively. The solid was refluxed in 93% ethanol (350ml) and then left to stand for 48 hours. The precipitate was filtered off and washed with ethanol and ether to give 4.8g of a white solid, which was analyzed by chiral HPLC to give two peaks, which were 9% and 91% of the total area of the two peaks, respectively. Reflux with 50% ethanol (48ml) and rest for an additional 48 hours. The precipitate was filtered off and washed with ethanol and ether to give 2.7g of a white solid, which upon analysis by chiral HPLC gave two peaks, 3% and 97% of the total area of the two peaks respectively. The solid was refluxed in 50% ethanol (22ml) and then left for 48 hours. The precipitate was filtered off and washed with ethanol and ether to give 1.6g of a white solid, which was analyzed by chiral HPLC to give two peaks, which were 1% and 99% of the total area of the two peaks, respectively. The resulting diastereomerically enriched salt was dissolved in distilled water (20ml) and treated with acetic acid (0.35ml, 6.4mmol) for 15 min. The resulting precipitate was filtered off, washed with water, acetone and ether and then dried under vacuum at 80 ℃ to give 1.1g of a white solid. This was dissolved in pure ethanol (55ml) and concentrated hydrochloric acid (11.5N, 11ml) was added to give a yellow solution which was stirred at room temperature for 68 hours. The resulting precipitate was filtered off and washed with water, ethanol and diethyl ether, then dried under vacuum at 80 ℃ to yield 770mg of enantiomerically enriched 5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7] -indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione. Analysis by Chiral HPLC (Chiral-AGP column, gradient elution with 2 to 5% acetonitrile in 10mM phosphate buffer pH6.9, elution peaks at 15 and 20 min) indicated an enantiomeric excess of 98%. The above procedure was carried out again using D- (+) -alpha-methylbenzylamine instead of L- (-) -alpha-methylbenzylamine. This gives 5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': the other enantiomer of 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione.

Preparation example 3: 5, 12-diethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was prepared in analogy to preparation 1, except that in step 1.a. 7-ethylcamptothecin (Sawada et al, medical chemistry Notification 39: 2574(1991)) was used instead of camptothecin. The title compound was obtained as a bright yellow solid, m.p. > 270 ℃. NMR-1H (DMSO): 0.92(t, 3H); 1.39(t, 3H); 1.93(q, 2H); 3.08(d, 2H); 3.25(q, 2H); 3.51(d, 2H); 5.32(s, 2H); 5.52(dd, 2H); 7.42(s, 1H); 7.76(t, 1H); 7.89(t, 1H); 8.18(d, 1H); 8.32(d, 1H). NMR-C13 (DMSO): 8.46 of; 14.15 of; 22.42, respectively; 36.50; 42.54, respectively; 49.95; 61.45, respectively; 73.35, respectively; 99.68; 122.61, respectively; 124.27, respectively; 126.76, respectively; 127.70, respectively; 128.27, respectively; 129.92, respectively; 130.18, respectively; 145.17, respectively; 145.82, respectively; 148.57, respectively; 152.15, respectively; 155.89, respectively; 159.26, respectively; 172.08.

preparation example 4: 5-ethyl-9, 10-difluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

a.2-Ethyl-2- (2-methoxy-4-pyridinyl) -1, 3-dioxolane

Water was azeotropically distilled off with a Dean Stark apparatus from a mixture of 2-chloro-4-propionylpyridine (10g, 59mmol), ethylene glycol (20ml) and p-toluenesulfonic acid (250mg) in toluene (150ml) from Lamatina, J.L., J.Chem.20, p.553 (1983). The solvent was then removed under reduced pressure, the acid was neutralized with saturated aqueous sodium bicarbonate (100ml) and the product was extracted with diethyl ether. The combined ether extracts were washed with brine, dried over sodium sulfate and evaporated to give 13.3g (96%) of the crude carbonyl-protected product, which was heated under reflux with 3 equivalents of sodium methoxide in acetonitrile until the reaction was complete (detected by thin layer chromatography: silica, tert-butyl methyl ether/hexane (TBMO/HX) 50/50). The acetonitrile solution was then filtered and evaporated. The residue was dissolved in diethyl ether, washed with water and brine, dried over sodium sulfate and evaporated to give a brown oil which was distilled (70-75 ℃, 0.04 mbar); 10.7g (overall yield 81%) of product (F) are obtained as a clear oil.

2-Ethyl-2- (3-hydroxymethyl-2-methoxy-4-pyridinyl) -1, 3-dioxolane

Tert-butyllithium (1.7M pentane solution, 100ml, 170mmol) was added dropwise to a solution of bromo * (13ml, 85mmol) in anhydrous tetrahydrofuran (300ml) at-78 ℃ under argon. The resulting white precipitate was stirred at-78 ℃ for 1 hour, then 2-ethyl-2- (2-methoxy-4-pyridyl) -1, 3-dioxolane (10g, 44.8mmol) was added and the reaction mixture was stirred at-78 ℃ for 15 minutes, at 0 ℃ for 1 hour and at room temperature for 1 hour. It was cooled to-78 ℃ again, anhydrous N, N-dimethylformamide (100mmol) was added and the reaction mixture was warmed to room temperature and stirred for 16 h before complete reaction of the starting materials was indicated by thin layer chromatography (silica, TBMO/HX: 50/50). The reaction was stopped with saturated ammonium chloride and the reaction mixture was extracted with diethyl ether (200ml, 50ml, 50 ml). The combined extracts were dried over sodium sulfate and evaporated to give a yellow oil which was purified by column chromatography (silica, TBMO/HX: 0/100 to 5/95 eluting the * derivative, then 20/80 to 50/50 eluting the product) to give the intermediate aldehyde (7 g). The aldehyde was dissolved in methanol (100ml) and treated with sodium borohydride (5g, 132mmol) and the resulting mixture was stirred until controlled analysis by thin layer chromatography indicated complete reaction of the intermediate aldehyde (about 1 hour). The solvent was then evaporated off and the residue was dissolved in ether, washed with water and brine, dried and the solvent was evaporated off. The residue was purified by column chromatography (silica, TBMO/HX: 10/90 to 50/50) to give 7G (62% overall) of product (G) as a yellow oil.

C.2- (3-benzyloxymethyl-2-methoxy-4-pyridyl) -2-ethyl-1, 3-dioxolane

A solution of 2-ethyl-2- (3-hydroxymethyl-2-methoxy-4-pyridine) -1, 3-dioxolane (7g, 30mmol) and benzyl chloride (5ml, 45mol) in anhydrous tetrahydrofuran (50ml) was added dropwise to a suspension of sodium hydride (80% in mineral oil, 1.85g, 61mmol) in anhydrous tetrahydrofuran (100ml) and the reaction mixture was kept under reflux for 16 hours. The reaction mixture was then cooled to room temperature, quenched with water (50ml) and concentrated under reduced pressure. The residue was dissolved in ether (150ml) and washed with water and brine, dried and evaporated. Purification by column chromatography (silica, TBMO/HX: 5/95 to 20/80) gave 9g (87%) of benzyl-protected product (H) as a clear oil.

D.1- (3-benzyloxymethyl-2-methoxy-4-pyridinyl) -propan-1-one

2- (3-benzyloxymethyl-2-methoxy-4-pyridyl) -2-ethyl-1, 3-dioxolane (9g, 27mmol) was treated with trifluoroacetic acid (10ml) and water (5ml) in a hot bath at a temperature of 120 ℃ for 3 hours. The reaction mixture was concentrated under reduced pressure and the residual traces of acid were neutralized by addition of saturated aqueous sodium bicarbonate. Extraction with diethyl ether followed by purification by column chromatography (silica, TBMO/HX: 10/90) gave 5.5g (70%) of product (I).

Beta-ethyl-beta-hydroxy-beta- (3-benzyloxymethyl-2-methoxy-4-pyridinyl) -propionic acid tert-butyl ester

Tert-butyl bromoacetate (13ml, 80mmol) was added dropwise to a suspension of zinc (5.3g, 80mmol, activated with 6N HCl for 10 seconds) under reflux in anhydrous tetrahydrofuran (60ml) which was washed successively with water to neutral pH, acetone and diethyl ether. After the end of the dropwise addition, the reaction medium is maintained under reflux for a further 10 minutes. Then, a solution of 1- (3-benzyloxymethyl-2-methoxy-4-pyridyl) -propan-1-one (5.8g, 20mmol) in anhydrous tetrahydrofuran (20ml) was added and the reaction mixture was stirred under reflux for another 1 hour. The reaction was stopped at 0 ℃ with saturated aqueous ammonium chloride (100ml) and the reaction mixture was extracted with diethyl ether. The combined extracts were dried over sodium sulfate and evaporated to give a yellow oil which was purified by column chromatography (silica, TBMO/HX: 5/95 to 10/90) to give tert-butyl ester (J) (7g, 95%) as a clear oil.

Beta-ethyl-beta-hydroxy-beta- (3-hydroxymethyl-2-methoxy-4-pyridinyl) -propionic acid tert-butyl ester

Beta-ethyl-beta-hydroxy-beta- (3-benzyloxymethyl-2-methoxy-4-pyridyl) -propionic acid tert-butyl ester (1g, 2.5mmol) was subjected to hydrogenolysis at normal pressure and room temperature using 5% palladium on carbon as a catalyst (50mg) and pure ethanol as a solvent (10 ml). Upon termination of the reaction (6 hours), the catalyst was filtered off and the solvent was evaporated off, yielding 0.7g (90%) of product (K) which was pure enough for the subsequent reaction.

g.5-Ethyl-1, 5-dihydro-5-hydroxy-9-methoxy-oxepino [3, 4-c ] pyridin-3 (4H) -one

beta-Ethyl-beta-hydroxy-beta- (3-hydroxymethyl-2-methoxy-4-pyridinyl) -propionic acid tert-butyl ester (8.8g, 28mmol) was treated with trifluoroacetic acid (30ml) for 3 hours at room temperature. The volatiles were evaporated and the residue was purified by column chromatography (silica, dichloromethane/methanol: 100/0 to 98/2) to give a clear oil which, after treatment with toluene, gave 5.9g (89%) of product (L) as white crystals m.p.97-98 ℃.

h.5-Ethyl-1, 5-dihydro-5-hydroxy-oxepino [3, 4-c ] pyridine-3, 9(4H, 8H) -dione

5-Ethyl-1, 5-dihydro-5-hydroxy-9-methoxy-oxepino [3, 4-c ] pyridin-3 (4H) -one (0.5g, 2.1mmol) was heated under reflux in 1N hydrochloric acid (20ml) for 9 hours. The reaction mixture was concentrated under reduced pressure and the residue was dried by addition and evaporation of toluene and then left overnight under reduced pressure in the presence of phosphorus pentoxide. The resulting oil was dissolved in anhydrous acetonitrile (5ml) and stirred under argon atmosphere for 24 hours. The precipitate was filtered off and dried to yield 0.23g (49%) of a white solid (M), m.p.118-119 ℃.

I.2-chloro-6, 7-difluoro-3-quinoline-methanol

Using Meth-Cohn and colleagues at j.chem.soc.perkin trans.i. page 1520 (1981); the method described by Meth-Cohn, J.chem.Soc.Perkin Trans.I. page 2509 (1981). 3, 4-difluoro-N-acetylaniline (38g, 22mmol) was added to Vilsmeyer's reagent prepared by dropwise addition of phosphoryl oxychloride (103ml, 1.1mmol) to anhydrous dimethylformamide (34ml, 44mmol), cooled with a water/ice bath and stirred under argon atmosphere for 0.5 h. The resulting mixture was heated at 70 ℃ for 16 hours. After cooling to room temperature, the reaction mixture was added to a mixture of ice and water (400ml) and stirring was continued for 2 hours, then filtered and washed successively with water, ethanol and diethyl ether to prepare 9g of 2-chloro-6, 7-difluoroquinoline-3-carbaldehyde as a yellow solid, m.p.222-224 ℃. This intermediate was treated with sodium borohydride (2g, 52mmol) in methanol (400ml) at room temperature for 0.5 h, then excess reagents were destroyed by addition of acetic acid (2 ml). The solvent was removed under reduced pressure, the residue was added to an ethyl acetate solution and washed successively with dilute sodium bicarbonate, water and saturated aqueous sodium chloride solution. Dry over sodium sulfate, filter and concentrate the organic phase. The resulting solid was recrystallized from 1, 2-dichloroethane to give 8g of 2-chloro-6, 7-difluoro-3-quinoline-methanol as a beige solid.

j.5-Ethyl-8- (2-chloro-6, 7-difluoro-3-quinolinylmethyl) -1, 5-dihydro-5-hydroxy-oxepino [3, 4-c ] pyridine-3, 9(4H, 8H) -dione

Diethyl azodicarboxylate (570 μ l, 3.6mmol) was added dropwise over 5 min to a solution of 5-ethyl-1, 5-dihydro-5-hydroxy-oxepino [3, 4-c ] pyridine-3, 9(4H, 8H) -dione (400mg, 1.79mmol), the compound obtained in step 4.i. (770mg, 2.23mmol) and triphenylphosphine (934mg, 3.58mmol) in anhydrous N, N-dimethylformamide (45ml), and the resulting mixture was stirred at room temperature under argon atmosphere for 16H. The reaction mixture was then concentrated under reduced pressure and the residue was dissolved in diethyl ether (100 ml). The resulting solution was washed with brine (4X 50ml), dried over sodium sulfate and evaporated. The residue is chromatographed by column chromatography (silica, dichloromethane/methanol: 99/1 to 98/2) to give 650mg (66%) of product (O) as a white solid, m.p.165-167 ℃.

4, k.5-ethyl-9, 10-difluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

5-Ethyl-8- (2-chloro-6, 7-difluoro-3-quinolinylmethyl) -1, 5-dihydro-5-hydroxy-oxepino [3, 4-c ] pyridine-3, 9(4H, 8H) -dione (600mg, 1.1mmol), tetrabutylammonium bromide (352mg, 1.1mmol), sodium acetate (359mg, 4.4mmol) and palladium acetate II (98mg, 0.43mmol) were dissolved in anhydrous acetonitrile (40ml) and heated at 90 ℃ under argon for 16 hours. After cooling to room temperature, a white precipitate separated from the red solution. The precipitate was filtered off and dried under reduced pressure. This crude product was suspended in water, filtered and dried over phosphorus pentoxide under reduced pressure to give 250mg of the title compound as an off-white solid, m.p. > 250 ℃. NMR-1H (DMSO): 0.91(t, 3H); 1.87(m, 2H); 3.08(d, 1H); 3.51(d, 1H); 4.45(s, 4H); 5.19(s, 2H); 5.47(dd, 2H); 6.02(se, 1H); 7.33(s, 1H); 7.54(s, 1H); 7.55(s, 1H); 8.43(s, 1H). NMR-C13 (DMSO): 8.43; 36.47, respectively; 42.54, respectively; 50.52; 61.43; 64.43 (2C); 73.31, respectively; 99.07, respectively; 112.27, respectively; 113.14; 122.00, respectively; 124.24, respectively; 128.18, respectively; 129.74, respectively; 144.59, respectively; 145.01, respectively; 145.33, respectively; 147.63; 150.88; 155.88, respectively; 159.23, respectively; 172.07.

preparation example 5: 5-ethyl-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': 6, 7-indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

10-benzyloxy-5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7] indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione (370mg, 0.79 mmol). Once the reaction had stopped (16 h), dichloromethane (50ml) and methanol (50ml) were added to the reaction mixture, the catalyst was filtered off and the volatiles were evaporated under reduced pressure, thus obtaining the title crude product containing traces of trifluoroacetic acid. Traces of trifluoroacetic acid were removed by co-distillation with 1, 4-dioxane. The product was obtained as an orange solid, m.p.150 ℃ with sufficient purity for subsequent syntheses. NMR-1H (DMSO): 0.89(t, 3H); 1.85(q, 2H); 3.02(d, 1H); 3.45(d, 1H); 5.19(s, 2H); 5.37(d, 1H); 5.50(d, 1H); 5.98(se, 1H); 7.26(s 1H); 7.31(s, 1H); 7.40(d, 1H); 8.00(d, 1H); 8.42(s, 1H); 10.32(s, 1H). NMR-C13 (DMSO): 8.47; 36.50; 42.61, respectively; 50.57, respectively; 61.46; 73.35, respectively; 98.84; 109.02, respectively; 121.83, respectively; 123.18, respectively; 129.50 of the total weight of the powder; 129.85, respectively; 130.12, respectively; 130.80, respectively; 143.39, respectively; 145.10, respectively; 149.69, respectively; 155.97, respectively; 156.82, respectively; 159.30, respectively; 172.11.

preparation example 6: 5-ethyl-9-fluoro-4, 5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3-fluoro-4-methoxyaniline. NMR-1H (DMSO): 0.89(t, 3H); 1.85(q, 2H); 3.08(d, 1H); 3.49(d, 1H); 4.00(s, 3H); 5.25(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.00(s, 1H); 7.32(s, 1H); 7.72(d, 1H); 7.91(d, 1H); 8.58(s, 1H). NMR-C13 (DMSO): 8.43; 36.48, respectively; 42.51; 50.68; 56.60, respectively; 61.42, respectively; 73.29; 99.25; 108.68, respectively; 113.52, respectively; 122.23, respectively; 126.33, respectively; 129.99, respectively; 130.30, respectively; 143.79, respectively; 144.70, respectively; 148.42, respectively; 151.18, respectively; 153.19, respectively; 155.81, respectively; 159.20, respectively; 172.06.IR (KBr): 1259; 1503; 1602; 1737.

preparation example 7: 9-chloro-5-ethyl-4, 5-dihydro-5-hydroxy-10-methyl-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3-chloro-4-methoxyaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 2.55(s, 3H); 3.07(d, 1H); 3.45(d, 1H); 5.25(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.39(s, 1H); 8.10(s, 1H); 8.20(s, 1H); 8.60(s, 1H). NMR-C13 (DMSO): 8.43; 20.20; 36.47, respectively; 42.49 of the total weight of the powder; 50.67, respectively; 61.41; 73.28, respectively; 99.87; 122.82, respectively; 126.98 of the total weight of the mixture; 127.99, respectively; 129.60, respectively; 130.53, respectively; 131.08 of the total weight of the mixture; 135.64, respectively; 136.56, respectively; 144.39, respectively; 147.11, respectively; 153.10, respectively; 155.85, respectively; 159.18, respectively; 172.03.IR (KBr): 1208; 1479; 1606; 1656; 1724.

preparation example 8: 8-Ethyl-2, 3, 8, 9-tetrahydro-8-hydroxy-10H, 12H- [1, 4] dioxino [2, 3-g ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

This compound was obtained as a yellow solid with m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3, 4-ethylenedioxyaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.47(d, 1H); 5.25(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.39(s, 1H); 8.15(q, 1H); 8.25(q, 1H); 8.68(s, 1H). NMR-C13 (DMSO): 8.41; 36.45 of; 42.48; 50.68; 61.40; 73.25; 99.92; 114.44, respectively; 115.42, respectively; 115.58, respectively; 122.96, respectively; 125.52; 130.56, respectively; 131.46, respectively; 144.21, respectively; 145.25, respectively; 142.36, respectively; 153.41, respectively; 155.85, respectively; 159.15, respectively; 172.00.IR (KBr): 1266; 1512; 1581; 1618; 1751.

preparation example 9: 7-ethyl-7, 8-dihydro-7-hydroxy-9H, 11H- [1, 3] dioxolo [4, 5-g ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-9, 12(14H) -dione

This compound was obtained as a cream solid, m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3, 4-methylenedioxyaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 5.20(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.00(s, 1H); 6.30(s, 2H); 7.30(s, 1H); 7.49(d, 2H); 8.45(s, 1H). NMR-C13 (DMSO): 8.43; 36.49; 42.56; 50.58, respectively; 61.42, respectively; 73.31, respectively; 98.87; 102.75, respectively; 103.33, respectively; 104.92; 121.76, respectively; 125.74, respectively; 128.59; 130.33, respectively; 145.08, respectively; 146.69, respectively; 148.78, respectively; 150.19, respectively; 151.49, respectively; 155.90, respectively; 159.24; 172.08.IR (KBr): 1248; 1459; 1606; 1731.

preparation example 10: 9-chloro-5-ethyl-4, 5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a white solid, m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3-chloro-4-methoxyaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 4.01(s, 3H); 5.22(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.02(s, 1H); 7.31(s, 1H); 7.68(s, 1H); 8.20(s, 1H); 8.55(s, 1H). NMR-C13 (DMSO): 8.22; 36.27, respectively; 42.30, respectively; 50.48; 56.69, respectively; 61.23, respectively; 73.08, respectively; 99.16, respectively; 107.44, respectively; 122.16; 127.12, respectively; 128.12, respectively; 129.25, respectively; 130.02; 130.53, respectively; 143.29, respectively; 144.37, respectively; 151.12; 153.29, respectively; 155.71, respectively; 158.98, respectively; 171.84.IR (KBr): 1056; 1256; 1483; 1592; 1657; 1747.

preparation example 11: 5-ethyl-4, 5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 4-methoxyaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 3.95(s, 3H); 5.28(s, 2H); 5.40(d, 1H); 5.51(d, 1H); 6.00(s, 1H); 7.38(s, 1H); 7.51(d, 2H); 8.07(d, 1H); 8.55(s, 1H). NMR-C13 (DMSO): 8.45 of; 36.48, respectively; 42.51; 50.64, respectively; 55.92; 61.42, respectively; 73.33; 99.01; 106.49, respectively; 122.02, respectively; 123.19, respectively; 129.59, respectively; 130.20, respectively; 130.43, respectively; 144.17, respectively; 144.94, respectively; 150.40, respectively; 155.92, respectively; 158.31, respectively; 159.26, respectively; 172.07.IR (KBr): 1251; 1604; 1655; 1735.

preparation example 12: 9, 11-dichloro-5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid with m.p. > 250 ℃ by starting from 3, 5-dichloroaniline according to the method of preparation 4, steps 4i, 4j and 4 k. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 5.30(s, 2H); 5.41(d, 1H); 5.55(d, 1H); 6.08(s, 1H); 7.41(s, 1H); 8.05(s, 1H); 8.21(s, 1H); 8.91(s, 1H). NMR-C13 (DMSO): 8.39; 36.45 of; 42.51; 51.03; 61.39; 73.25; 100.62 of; 123.55, respectively; 124.63, respectively; 127.60, respectively; 128.08, respectively; 128.56, respectively; 132.06, respectively; 132.19; 134.53, respectively; 143.77, respectively; 148.80; 154.88, respectively; 155.82, respectively; 159.13, respectively; 171.98.IR (KBr): 1064; 1275; 1586; 1651; 1743.

preparation example 13: 5-ethyl-9-fluoro-4, 5-dihydro-5-hydroxy-10-methyl-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 3-fluoro-4-methylaniline. NMR-1H (DMSO): 0.89(t, 3H); 1.85(q, 2H); 2.49(s, 3H); 3.08(d, 1H); 3.49(d, 1H); 5.21(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.39(s, 1H); 7.87(d, 1H); 8.05(d, 1H); 8.61(s, 1H). NMR-C13 (DMSO): 8.40; 15.14 of the total weight of the mixture; 36.45 of; 42.52, respectively; 50.60 parts of; 61.41; 73.28, respectively; 99.71; 112.00, respectively; 122.66, respectively; 125.38, respectively; 127.66, respectively; 129.59, respectively; 130.28, respectively; 144.49, respectively; 147.88, respectively; 152.88, respectively; 155.85, respectively; 159.18, respectively; 162.25, respectively; 172.02.IR (KBr): 1054; 1580; 1651; 1760.

preparation example 14: 5-ethyl-10-fluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a white solid with m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 4-fluoroaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 5.29(s, 2H); 5.39(d, 1H); 5.55(d, 1H); 6.30(s, 1H); 7.39(s, 1H); 7.80(q, 1H); 7.99(q, 1H); 8.23(q, 1H); 8.68(s, 1H). NMR-C13 (DMSO): 8.40; 36.46, respectively; 42.48; 50.66; 61.41; 73.31, respectively; 99.68; 111.83, respectively; 122.75, respectively; 128.93, respectively; 130.93, respectively; 131.22, respectively; 131.93, respectively; 144.46, respectively; 145.27, respectively; 152.60, respectively; 155.89, respectively; 159.21, respectively; 172.04.IR (KBr): 1209; 1589; 1659; 1739.

preparation example 15: 10-chloro-5-ethyl-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

This compound was obtained as a yellow solid with m.p. > 250 ℃ following the procedure of preparation 4, steps 4i, 4j and 4k, starting from 4-chloroaniline. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.47(d, 1H); 5.25(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.39(s, 1H); 7.89(d, 1H); 8.19(d, 1H); 8.29(s, 1H); 8.67(s, 1H). NMR-C13 (DMSO): 8.40; 36.46, respectively; 42.47; 50.70, respectively; 61.42, respectively; 73.31, respectively; 100.00; 122.96, respectively; 127.31, respectively; 127.42, respectively; 128.87, respectively; 131.11, respectively; 132.12, respectively; 144.34, respectively; 146.53, respectively; 153.38, respectively; 155.88, respectively; 159.20, respectively; 172.04.IR (KBr): 1069; 1483; 1606; 1741.

preparation example 16: 9-chloro-5-ethyl-10-fluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

The reaction was started from 4-chloro-3-fluoroaniline following the procedure of preparation 4 steps 4i, 4j and 4k to give this compound as a yellow solid with m.p. > 250 ℃. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.07(d, 1H); 3.45(d, 1H); 5.25(s, 2H); 5.39(d, 1H); 5.51(d, 1H); 6.05(s, 1H); 7.40(s, 1H); 8.20(d, 1H); 8.40(d, 1H); 8.68(s, 1H). NMR-C13 (DMSO): 8.38; 36.47, respectively; 42.58, respectively; 50.71, respectively; 61.40; 73.26, respectively; 99.99 of the total weight of the steel; 113.59, respectively; 123.09, respectively; 124.28, respectively; 127.74, respectively; 130.64, respectively; 131.31, respectively; 144.13, respectively; 145.08, respectively; 153.57, respectively; 154.13, respectively; 155.84, respectively; 156.61, respectively; 159.14, respectively; 172.00.IR (KBr): 1488; 1583; 1655; 1743.

preparation example 17: 5, 12-diethyl-9-fluoro-4, 5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

A.5-fluoro-4-methoxy-2-propionylaniline

This product was obtained according to Sugasawa T, Toyoda T, Adachi M, Sasakura K, J.Am.Chem.Soc.100 (1978), pages 4842 and 4852. To a solution of 3-fluoro-4-methoxyaniline (20g, 142mmol) in dry dichloromethane (200ml) was added dropwise boron trichloride (1M in heptane, 156ml, 156mmol) under nitrogen at 0 ℃. The pink suspension thus obtained was stirred for 5 minutes, then propionitrile (33ml, 420mmol) was added dropwise and aluminium trichloride (20.8g, 156mmol) was added in small portions. The reaction medium is heated under reflux for 3 hours, cooled to 0 ℃ and hydrolyzed by careful addition of 2N hydrochloric acid (100ml) and then heated under reflux for 45 minutes. After cooling to 0 ℃, the resulting precipitate was filtered off, washed with dichloromethane and then recovered in water (300 ml). The aqueous phase was adjusted to basic pH, extracted with dichloromethane and then extracted with ethyl acetate. The organic phase is dried over magnesium sulfate and evaporated to give the crude product which is purified by column chromatography (silica, ethyl acetate/heptane: 1/99 to 20/80). 15.3g of a yellow solid are obtained. NMR-1H (CDCl)₃)：1.20(t，3H)；2.92(q，2H)；3.83(s，3H)；6.2(s，2H)；6.40(d，2H)；7.32(d，2H).IR(KBr)：857；1148；1240；1561；1583；1662.

b.4-Ethyl-7-fluoro-2-hydroxy-6-methoxy-3-quinolinecarboxylic acid ethyl ester

A solution of ethylmalonyl chloride (12.9ml, 100mmol) in dry acetonitrile (30ml) was added dropwise to a solution of 5-fluoro-4-methoxy-2-propionylaniline (15.3g, 77.5mmol) and triethylamine (13.9ml, 100mmol) in dry acetonitrile (110ml) under argon at 0 ℃. The temperature of the reaction medium is allowed to return to room temperature, a solution of sodium ethoxide (prepared from 1.8g of 78mmol of sodium in 80ml of ethanol) is added dropwise via a conduit under argon, and the reaction medium is then stirred at room temperature for 12 hours. The reaction mixture was poured into ice-cold water (100ml) and stirred for 2 hours, then the precipitate was filtered off and washed with water, ethanol and diethyl ether. 19.4g of a white solid was obtained. NMR-1H (DMSO): 1.25(m, 6H); 2.78(q, 2H); 3.92(s, 3H); 4.30(q, 2H); 7.15(d, 2H); 7.40(d, 2H); 11.93(s, 1H). IR (KBr): 786; 1083; 1410; 1521, a carrier gas supply device; 1644; 1725.

c.2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinecarboxylic acid ethyl ester

A suspension of ethyl 4-ethyl-7-fluoro-2-hydroxy-6-methoxy-3-quinolinecarboxylate (19.4g, 0.066mol) in phosphorus oxychloride (243ml) was heated under reflux for 6 hours. Phosphorus oxychloride was distilled off. The reaction mixture was poured into ice-cold water, which was then dissolved in dichloromethane. The organic phase is washed with water and then with a saturated sodium chloride solution. The organic phase was dried over magnesium sulfate and the solvent was evaporated. The residue was suspended in diethyl ether and the unreacted starting material (4g) was filtered off. The filtrate was evaporated and the residue was purified by column chromatography (silica, ethyl acetate/heptane: 5/95 to 20/80). 10.9g of a white solid was obtained. NMR-1H (DMSO): 1.30(t, 3H); 1.39(t, 3H); 3.08(q, 2H); 4.09(s, 3H); 4.49(q, 2H); 7.64(d, 2H); 7.86(d, 2H). IR (KBr): 865; 1016; 1082; 1190, a pharmaceutically acceptable carrier; 1224 of the base material; 1253; 1272; 1508; 1571; 1732.

d.2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinemethanol

Diisobutylaluminum hydride (1M in dichloromethane, 65ml, 65mmol) was added dropwise to a solution of ethyl 2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinecarboxylate (10.8g, 35mmol) in anhydrous dichloromethane (200ml) at room temperature under an inert atmosphere, followed by heating at 40 ℃ for 4 hours. After cooling to 0 ℃, 20% aqueous solution of sodium potassium tartrate (105ml) and dichloromethane (200ml) were carefully added and the reaction mixture was stirred for 1 hour, then decanted and washed 3 times with water. The organic phase was dried over magnesium sulfate and the solvent was evaporated. The residue is purified by column chromatography (silica, ethyl acetate/heptane: 5/95 to 50/50). 6g of a white solid are obtained. NMR-1H (DMSO): 1.28(t, 3H); 3.25(q, 2H); 4.04(s, 3H); 4.77(d, 2H); 5.27(t, 1H); 7.55(d, 2H); 7.73(d, 2H). IR (KBr): 840; 864; 1023; 1232, and; 1267; 1317; 1444, respectively; 1511; 1569.

17, e.5, 12-diethyl-9-fluoro-4, 5-dihydro-5-hydroxy-10-methoxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

2-chloro-4-ethyl-7-fluoro-6-methoxy-3-quinolinemethanol was coupled with compound (M) as described in preparation 4, step 4.j. The resulting coupled product is cyclized as described in step 4.k. A yellow solid was obtained, m.p. > 275 ℃. NMR-1H (CF3 COOD): 1.07(m, 3H); 1.62(m, 3H); 2.27(m, 2H); 3.44(d, 1H); 3.54(m, 2H); 3.91(d, 1H); 4.25(s, 3H); 5.60(d, 1H); 5.74(s, 2H); 5.98(d, 1H); 7.85(m, 1H); 8.16(m, 1H); 8.31(s, 1H). NMR-C13(CF3 COOD): 9.03; 14.20 of the total weight of the powder; 26.68; 38.77; 43.98; 53.79; 58.27; 64.73; 77.93; 106.85, respectively; 109.24, respectively; 110.15 of the total weight of the alloy; 128.99, respectively; 129.20, respectively; 131.61, respectively; 137.32, respectively; 141.23, respectively; 144.13, respectively; 154.79, respectively; 158.32, respectively; 160.25, respectively; 160.81, respectively; 179.30.IR (KBr): 1013; 1068; 1265; 1466; 1514; 1601; 1655; 1748.

preparation example 18: 5-ethyl-4, 5-dihydro-5-hydroxy-12-methyl-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

2-chloro-4-methyl-3-quinolinemethanol was prepared from 2-acetanilide using the methods described in preparations 17.b., 17.c., and 17.d. The latter was coupled with compound (M) according to the procedure of preparation 4, step 4.j. The resulting coupled product is cyclized as described in step 4.k. A yellow solid was obtained, m.p. > 260 ℃. NMR 1h (dmso): 0.87(t, 3H); 1.87(q, 2H); 2.78(s, 3H); 2.80(d, 1H); 3.55(d, 1H); 5.27(s, 2H); 5.42(d, 1H); 5.52(d, 1H); 6.04(s, 1H); 7.39(s, 1H); 7.75(t, 1H); 7.88(t, 1H); 8.13(d, 1H); 8.25(d, 1H). NMR-C13 (DMSO): 8.23; 36.26; 42.36; 62.00; 73.11; 78.65 of; 79.13, respectively; 79.25; 99.52; 122.36, respectively; 124.30, respectively; 127.67; 129.54 of the total weight of the powder; 129.55, respectively; 129.56, respectively; 140.11, respectively; 145.06, respectively; 148.07, respectively; 152.00, respectively; 155.79, respectively; 159.09, respectively; 171.89.IR (KBr): 1649; 1751, preparing a mixture of water and an organic solvent; 3404.

preparation example 19: 10-benzyloxy-5-ethyl-9-fluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

The procedure described in preparation 4.i. was used to prepare 2-chloro-7-fluoro-6-methoxy-quinoline-3-carbaldehyde from 3-fluoro-4-methoxy-acetanilide, which was treated with excess boron tribromide in dichloromethane for 24 hours at room temperature. The resulting 2-chloro-7-fluoro-6-hydroxy-quinoline-3-carbaldehyde was O-benzylated in dimethylformamide in the presence of benzyl bromide and potassium carbonate to produce 6-benzyloxy-2-chloro-7-fluoro-quinoline-3-carbaldehyde, which was reduced with sodium borohydride in methanol to produce the corresponding quinoline methanol. The latter was coupled with compound (M) as described in preparation 4, step 4.j. The resulting coupled product is cyclized as described in step 4.k. A yellow solid was obtained, m.p. > 275 ℃. NMR-1H (DMSO): 0.86(t, 3H); 1.85(q, 2H); 3.05(d, 1H); 5.25(s, 2H); 5.37(s, 2H); 5.45(dd, 2H); 6.05(s, 1H); 7.4-7.6(m, 5H); 7.88(d, 1H); 7.95(d, 1H); 8.56(s, 1H).

Preparation example 20: 5-ethyl-9-fluoro-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

The compound (0.79mmol) of production example 19 was dissolved in trifluoroacetic acid (15ml), which was treated with hydrogen gas and 10% palladium on carbon (60mg) as a catalyst. A yellow solid was obtained, m.p. > 275 ℃. NMR-1H (DMSO): 0.86(t, 3H); 1.85(q, 2H); 3.05(d, 1H); 5.25(s, 2H); 5.37(s, 2H); 5.45(dd, 2H); 6.05(s, 1H); 7.8(d, 1H); 7.90(d, 1H); 8.56(s, 1H).

The above preparation examples serve as a basis for the description of the invention by the following examples.

Example 1:

5-ethyl-9, 10-difluoro-4, 5-dihydro-5- (2-amino-1-oxoethoxy) -1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

5-Ethyl-9, 10-difluoro-4, 5-dihydro-5- (2- (tert-butoxycarbonylamino) -1-oxoethoxy) -1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione hydrochloride

Reacting 5-ethyl-9, 10-difluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6,7]Indolizino [1, 2-b ]]Quinoline-3, 15(4H, 13H) -dione (200mg, 0.526mmol, from preparation 4), N-Boc-glycine (185mg, 1.051mmol) and a catalytic amount of 4-dimethylaminopyridine (20mg) in dry pyridine (10ml) was treated with dicyclohexylcarbodiimide (239mg, 1.16mmol) at 0 ℃ under argon and stirred at room temperature for 48H. The volatiles were removed in vacuo and the residue was purified by chromatography (silica, 1% methanol in chloroform) to give the desired intermediate (40mg, 14%) as a yellow solid. NMR-1H (CDCl)₃)：1.20(t，3H)；1.38(s，9H)；1.40-1.70(m，2H)；3.10(d，1H)；4.00(d，2H)；4.30(d，1H)；5.00(t，1H)；5.20(s，2H)；5.30-5.90(dd，2H)；7.20(s，1H)；7.50-8.10(m，2H)；8.30(s，1H).

5-ethyl-9, 10-difluoro-4, 5-dihydro-5- (2-amino-1-oxoethoxy) -1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione hydrochloride

A solution of the intermediate obtained above (40mg, 0.072mmol) in dichloromethane (10ml) was maintained at 0 ℃ and dioxane saturated with hydrogen chloride (8ml) was added dropwise. The resulting yellow suspension was stirred for 2 hours, and then the volatiles were removed in vacuo. The residue was dissolved in water (5ml) and washed with dichloromethane (3X 30 ml). Freezing and lyophilizing the aqueous phase to prepare a pre-gelThe salt of phase (b) was a hygroscopic yellow solid (20mg, 50%). NMR-1H (CDCl)₃)：1.00(t，3H)；2.15(m，1H)；2.30(m，1H)；3.60(d，1H)；3.90(d，1H)；4.15(s，2H)；5.10(s，2H)；5.40(d，1H)；5.70(d，2H)；7.40(s，1H)；

7.80(m，2H)；8.50(s，1H).

Example 2:

5-ethyl-9, 10-difluoro-4, 5-dihydro-5- (2-amino-1-oxopropoxy) -1H-oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione

Following the procedure of example 1, the compound was purified from 5-ethyl-9, 10-difluoro-4, 5-dihydro-5-hydroxy-1H-oxepino [3 ', 4': 6, 7] indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione, and replacing N-Boc-glycine with N-Boc-b-alanine, and cleaving the Boc protecting group on the intermediate by treatment with trifluoroacetic acid in dichloromethane. The volatiles were evaporated in vacuo and the residue was dissolved in dichloromethane. The resulting solution was washed with dilute bicarbonate, dried and evaporated. A yellow solid was obtained.

Similar results were obtained for other compounds using the methods of examples 1 and 2. All camptothecin analogs are thus available in "prodrug" form.

Example 3:

1, 8-diethyl-8, 9-dihydro-8-hydroxy-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

Reacting 5-ethyl-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': a suspension of 6, 7] -indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione (84mg, prepared according to preparation 5) in acetic acid (2.5ml) was treated with 1, 3, 5-triethylhexahydrotriazine (0.5 ml). The reaction mixture was stirred at 70 ℃ for 30 minutes and then evaporated in vacuo. The residue was dissolved in ethanol, filtered and washed with diethyl ether. A solid was obtained, m.p. > 275 ℃. NMR-1H (DMSO): 0.87(t, 3H); 1.50(t, 3H); 1.85(q, 2H); 2.77(q, 2H); 3.05(d, 1H); 3.47(d, 1H); 4.37(s, 2H); 5.00(s, 2H); 5.22(s, 2H); 5.45(dd, 2H); 6.00(s, 1H); 7.34(s, 1H); 7.36(d, 1H); 7.93(d, 1H); 8.53(s, 1H). NMR-C13 (DMSO): 8.46 of; 13.48; 36.46, respectively; 42.49 of the total weight of the powder; 45.49, respectively; 46.44; 50.75; 61.43; 73.33; 82.06; 99.02; 112.90, respectively; 122.00, respectively; 122.98, respectively; 125.42, respectively; 127.04, respectively; 129.04, respectively; 130.20, respectively; 144.09, respectively; 144.97, respectively; 149.87, respectively; 152.92, respectively; 155.98, respectively; 172.07.IR (KBr): 1045; 1215; 1502; 1604; 1657, 1722.

Example 4:

8-ethyl-8, 9-dihydro-8-hydroxy-1-methyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

Reacting 5-ethyl-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': a suspension of 6, 7] -indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione (200mg, prepared according to preparation 5) in acetic acid (5ml) was treated with hexahydro-1, 3, 5-trimethyltriazine (110 mg). The reaction mixture was stirred at 70 ℃ for 30 minutes and then evaporated in vacuo. The residue was recovered with ethanol, filtered and washed with diethyl ether. A solid was obtained, m.p. > 275 ℃. NMR-1H (DMSO): 0.87(t, 3H); 1.85(q, 2H); 3.04(d, 1H); 3.48(d, 1H); 4.33(s, 2H); 4.93(s, 2H); 5.28(s, 2H); 5.45(dd, 2H); 6.01(s, 1H); 7.35(s, 1H); 7.38(d, 1H); 7.94(d, 1H); 8.49(s, 1H). NMR-C13 (DMSO): 8.46 of; 36.43, respectively; 37.85; 42.55, respectively; 48.68, respectively; 50.79, respectively; 61.43; 73.35, respectively; 83.82; 99.04; 112.49 of the total weight of the mixture; 122.04, respectively; 123.00, respectively; 125.46, respectively; 127.14, respectively; 129.07, respectively; 130.27, respectively; 144.99, respectively; 149.95, respectively; 152.46, respectively; 155.99, respectively; 172.09IR (KBr): 1047; 1058; 1219 of (a); 1246; 1295, 1439; 1504; 1604, 1655, 1735.

Example 5:

8-ethyl-8, 9-dihydro-8-hydroxy-1-benzyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

Reacting 5-ethyl-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': a suspension of 6, 7] -indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione (200mg, prepared according to preparation 5) in acetic acid (5ml) was treated with 1, 3, 5-tribenzylhexahydrotriazine (285 mg). The reaction mixture was stirred at 70 ℃ for 30 minutes and then evaporated in vacuo. The residue was recovered with ethanol, filtered and washed with diethyl ether. A solid was obtained, m.p. > 275 ℃. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.05(d, 1H); 3.47(d, 1H); 3.96(s, 2H); 4.33(s, 2H); 5.04(s, 2H); 5.17(s, 2H); 5.44(dd, 2H); 6.01(s, 1H); 7.38(m, 6H); 7.42(d, 1H); 7.97(d, 1H); 8.42(s, 1H). NMR-C13 (DMSO): 8.42; 19.96; 36.45 of; 42.51; 46.36, respectively; 50.78; 55.38, respectively; 61.39; 73.31, respectively; 99.00; 112.55, respectively; 122.01, respectively; 123.08 of the total weight of the mixture; 125.38, respectively; 127.09, respectively; 127.47, respectively; 128.70, respectively; 129.14, respectively; 130.35, respectively; 128.40, respectively; 139.19, respectively; 144.18, respectively; 149.99: 152.84; 155.92, respectively; 159.24; 172.05.IR (KBr): 1056; 1205; 1225; 1248; 1504; 1535; 1599; 1655; 1726.

example 6:

8-ethyl-8, 9-dihydro-4-fluoro-8-hydroxy-1-benzyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

Reacting 5-ethyl-9-fluoro-4, 5-dihydro-5, 10-dihydroxy-1H-oxepino [3 ', 4': a suspension of 6, 7] -indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione (200mg, prepared according to preparation 20) in acetic acid (5ml) was treated with 1, 3, 5-tribenzylhexahydrotriazine (285 mg). The reaction mixture was stirred at 70 ℃ for 30 minutes and then evaporated in vacuo. The residue was recovered with ethanol, filtered and washed with diethyl ether. A solid was obtained, m.p. > 250 ℃. NMR-1H (DMSO): 0.85(t, 3H); 1.85(q, 2H); 3.05(d, 1H); 3.48(d, 1H); 3.95(s, 2H); 4.45(s, 2H); 5.20(s, 4H); 5.45(d d, 2H); 6.05(s, 1H); 7.40(s, 7H); 7.90(d, 1H); 8.45(s, 1H). IR (KBr): 1248; 1451; 15001; 1598; 1657; 1727.

pharmacological study of the products of the invention

Topoisomerase 1-induced relaxation Activity assay of DNA

All reactions were carried out in 20. mu.l of a reaction buffer consisting of 50mM Tris-HCl (pH7.5), 50mM potassium chloride, 0.5mM dithiothreitol, 10mM magnesium chloride, 0.1mM ethylenediaminetetraacetic acid (EDTA), 30. mu.g/ml fetal bovine serum albumin, and 300ng of supercoiled pUC19(Pharmacia Biotech, Orsay, France) with or without a fixed concentration of the compound to be tested. All compounds tested were initially dissolved in dimethyl sulfoxide (DMSO) or water as appropriate for water soluble compounds, and additional dilutions were made with distilled water. The final concentration of DMSO does not exceed 1% (v/v). The reaction was initiated by addition of purified calf thymus DNA topoisomerase 1(Life Technologies/Gibco-BRL, Paisley, United kingdom) at 37 ℃ and was allowed to complete after 15 minutes. The reaction was stopped by adding 3. mu.l of a mixture containing 1% sodium dodecyl sulfate, 20mM EDTA and 500. mu.g/ml K protein kinase (Boehringer Mannheim, Meylan, France). After further incubation at 37 ℃ for 30 minutes, 2. mu.l of a loading buffer containing 10mM disodium hydrogen phosphate, 0.3% bromophenol blue and 16% Ficoll was added to the sample, and the sample was subjected to electrophoresis at 1V/cm for 20 hours on 1.2% agar gel in a buffer containing 36mM Tris-HCl (pH7.8), 30mM disodium hydrogen phosphate, 1mM EDTA and 2. mu.g/ml chloroquine. The gel was stained with 2. mu.g/ml ethidium bromide, photographed under UV light at 312nm with a charge coupled device (ccd) camera and the fluorescence intensity was measured with a bioProfil imaging analyser (Viber Lourmat, Lyon, France) to determine the percentage of DNA released.

In each experiment, supercoiled plasmid DNA alone or with topoisomerase 1 was cultured. The reaction was completed in 15 minutes. For each test compound or control (which will contain only the vector referred to as the control), the supercoiled plasmid DNA is incubated at the experimental maximum concentration of the test compound or control in the absence of the enzyme, or in the presence of the test compound at a concentration of 1. mu.M to 200. mu.M or in the presence of the control in the presence of the enzyme. As shown in table I, examples 3 to 6 inhibited the relaxing activity induced by topoisomerase 1 in a concentration-dependent manner.

TABLE 1

	Micromolar concentration
					10	50	100	200
	Examples
Camptothecin					88.7	62.4	52.9	46.9
	3	79.7	46.9	33.5					23.2
4					86.2	32.7	35.1	32.1
	5	56.2	30.4	28.0					24.2
6					55.6	39.9	38.9	30.0

Claims (10)

1.A compound, in its racemic or enantiomeric form or any mixture of these forms, or a pharmaceutically acceptable salt thereof, characterized in that it has the structure shown in formula (I):

R₁represents a lower alkyl group;

R₂、R₃and R₄Independently represents H, halogen, lower alkyl, (CH)₂)NR₆R₇、OR₆Or (CH)₂)[N＝X]Or is orR₃And R₄Together form a 3-or 4-membered chain, wherein the elements of the chain are selected from CH, CH₂O, S, N or NR₉；

R₅Represents H, lower alkyl, (CH)₂)NR₆R₇Or unsubstituted or substituted (CH) wherein the substituent is lower alkyl₂)[N＝X]；

R₆And R₇Independently represents H, lower alkyl or lower arylalkyl;

R₉represents lower alkyl or aralkyl;

R₁₈and R₁₉Independently represents H;

R₂₀represents H;

R_prepresents H or corresponds to the formula-C (O) -A-NR susceptible to cleavage₂₂R₂₃Wherein A represents a linear or branched alkylene group and R₂₂And R₂₃Independently represents H or lower alkyl; and is

[N＝X]Represents a 4-to 7-membered heterocyclic group, X represents the chain required to constitute said heterocyclic group and is selected from O, S, CH₂CH, N and NR₉；

Provided that R is_pIs a hydrogen atom, R₃And R₄Together form a 3 or 4 membered chain.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, characterized in that R₁Represents an ethyl group.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, characterized in that R₃And R₄To form an oxazine ring which may be substituted.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, characterized in that R_pAre groups that are easily cleaved.

5. A compound according to claim 3, characterized in that said compound is selected from the group consisting of:

-1, 8-diethyl-8, 9-dihydro-8-hydroxy-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

-8-ethyl-8, 9-dihydro-8-hydroxy-1-methyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

-8-ethyl-8, 9-dihydro-8-hydroxy-1-benzyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7 indolizino [1, 2-b ] quinoline-10, 13(15H) -dione

-8-ethyl-8, 9-dihydro-4-fluoro-8-hydroxy-1-benzyl-2H, 10H, 12H- [1, 3] oxazino [5, 6-f ] oxepino [3 ', 4': 6, 7] indolizino [1, 2-b ] quinoline-10, 13(15H) -dione.

6. The compound of claim 4, or a pharmaceutically acceptable salt thereof, characterized in that said compound is selected from the group consisting of:

5-ethyl-9, 10-difluoro-4, 5-dihydro-5- (2-amino-1-oxoethoxy) -1H-oxepino [3 ', 4': 6, 7] indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione;

5-ethyl-9, 10-difluoro-4, 5-dihydro-5- (2-amino-1-oxopropoxy) -1H-oxepino [3 ', 4': 6, 7] indolizino [1, 2-b ] quinoline-3, 15(4H, 13H) -dione.

7. Pharmaceutical composition containing at least one compound according to any one of claims 1 to 6 as active ingredient.

8. Use of a compound according to any one of claims 1 to 6 in the preparation of an anti-neoplastic medicament.

9. A process for the preparation of a compound of formula Ia corresponding to the product of formula I wherein R3 and R4 form an oxazine ring, as claimed in any one of claims 1, 3 or 5, characterized in that:

reacting a beta-hydroxy lactone compound of the formula D：

Wherein R is₃Is hydroxy, R₄Is H and R₁、R₂、R₁₈、R₁₉And R₂₀As defined above, with a primary amine under Mannich reaction conditions to give a β -hydroxy lactone compound of formula Ia

Wherein R is₁、R₂、R₄、R₉、R₁₈And R₁₉As defined above.

10. Claim 1 corresponds to wherein R_pA process for the preparation of a compound of formula Ib which is not a product of formula I having a hydrogen atom, characterized in that:

compounds of the formula DOr Ia

Using C (O) -A-N-R as described in claim 3₂₂R₂₃Acylation of derivatives of the radicals to prepare compounds in which R is_pBeta-hydroxy lactone compounds of formula I other than H.