CN1304380C - Benzo[a]henazin-11-carboxamide derivatives and their use as joint inhibitors of topomerase I and II - Google Patents

Abstract

A compound which is a benzo[a]phenazine-11-carboxamide derivative of formula (I) wherein each of R<1> to R<4>, which are the same or different, is selected from hydrogen, halogen, hydroxyl, C1-C6 alkoxy which is unsubstituted or substituted, heteroaryloxy, C1-C6 alkyl which is unsubstituted or substituted, nitro, cyano, azido, amidoxime, CO2R<10>, CON(R<12>)2, OCON(R<12>), SR<10>, SOR<11>, SO2(R11), SO2N(R<12>)2, N(R<12>)2, NR<10>SO2R<11>, N(SO2R<11>)2NR<10>(CH2)nCN, NR<10>COR<11>, OCOR<11> or COR<10>; each of R<5> to R<7>, which are the same or different, is selected from hydrogen, halogen, hydroxy, C1-C6 alkoxy, C1-C6 alkyl, SR<10> and N(R<12>)2; Q is C1-C6 alkylene which is unsubstituted or substituted by (i) C1-C6 alkyl which is unsubstituted or substituted, (ii) hydroxy, provided that the hydroxy group is not alpha to either of the N atoms adjacent to Q in formula (I), (iii) CO2R<10>, or (iv) CON(R<12>); R<8> and R<9>, which are the same or different, are each hydrogen or C1-C6 alkyl, or R<8> and R<9> together with the nitrogen atom to which they are attached form a saturated 5- or 6-membered N-containing heterocyclic ring which may include one additional heteroatom selected from O, N and S, or one of R<8> and R<9> is an alkylene chain optionally interrupted by O, N or S, which is attached to a carbon atom on the alkylene chain represented by Q to complete a saturated 5- or 6-membered N-containing heterocyclic ring as defined above; or a pharmaceutically acceptable salt thereof; with the proviso that at least one R<1> to R<4> is other than hydrogen. These compounds are inhibitors of topoisomerase I and/or topoisomerase II and can be used to treat tumours, including tumours which express MDR.

Description

Benzo[a]phenazine-11-amide derivatives and their use as combined inhibitors of topoisomerases I and II

The present invention relates to substituted benzo[a]phenazine-11-amides and derivatives thereof. These compounds are cytotoxic agents with proven topoisomerase I and topoisomerase II inhibition and the ability to prevent multidrug resistance mechanisms. They are therefore potential anticancer agents.

Topoisomerases are important cellular targets of many successful chemotherapeutic agents (Wang, Ann. Rev. Biochem, 65, 635-692, 1996) and in the regulation of DNA topology (required when cells divide and proliferate). regulation) important enzymes (Wang, loc. cit.). Drugs targeting topoisomerase II, such as doxorubicin and etoposide (Hande, Biophys. Acta 1400, 173-184, 1998), are widely used in the chemotherapy of cancer, and more recently those clearly Drugs targeting topoisomerase I (mainly camptothecin analogs) have had an important impact, an example being CPT-11 for the treatment of colon cancer (Dancey et al., Br. J. Cancer 74 , 327-338, 1996). More recently, topoisomerases have been shown to be therapeutic targets for antifungal, antibacterial and antiviral drugs (Chen et al., Rev. Pharmacol. Toxicol, 34, 191-218, 1994).

In addition to these compounds that specifically target topoisomerase I or II, several combined inhibitors of topoisomerase I and II have been identified and are also beneficial for the treatment of solid tumors. These compounds include intolicin (Riou et al., CancerRes.53, 5987-5993, 1993), DACA/XR5000 (Finlay et al., Eur. J. Cancer 32A, 708-714, 1996) and TAS-103 (Utsugi et al. People, J. Cancer Res, 88, 992-1002, 1997), these compounds are undergoing clinical evaluation. The advantages of combined inhibitors of topoisomerase I and II lie in their ability to avoid drug resistance and to target two key enzymes affecting DNA topology that are active at different points in the cell cycle.

A novel class of benzo[a]phenazine-11-amides has now been discovered as inhibitors of topoisomerase I and topoisomerase II. Therefore, the present invention provides a compound or a pharmaceutically acceptable salt thereof which is a benzo[a]phenazine-11-amide derivative of formula (I):

In the formula, R ¹ -R ³ are hydrogen and R ⁴ is selected from halogen, hydroxyl, unsubstituted or selected from N(R ¹² ) ₂ , CON(R ¹² ) ₂ , hydroxyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, cyano, CO ₂ R ¹⁰ , COR ¹⁰ , saturated 5- or 6-membered N-containing heterocyclic group, unsubstituted C ₁ -C ₆ alkoxy, heteroaryloxy, unsubstituted or substituted by a group of phenyl and phenyl substituted by one or more halogen atoms, or selected from hydroxy-C ₁ -C ₆ -alkyl , C ₁ -C ₆ -alkoxy, phenyl, N(R ¹² ) ₂ and hydroxyl groups substituted C ₁ -C ₆ alkyl, nitro, cyano, azido, amidoxime, CO ₂ R ¹⁰ , CON(R ¹² ) ₂ , OCON(R ¹² ) ₂ , SR ¹⁰ , SOR ¹¹ , SO ₂ R ¹¹ , SO ₂ N(R ¹² ) ₂ , N(R ¹² ) ₂ , NR ¹⁰ SO ₂ R ^11. N(SO ₂ R ¹¹ ) ₂ , NR ¹⁰ (CH ₂ ) _n CN, NR ¹⁰ COR ¹¹ , OCOR ¹¹ or COR ¹⁰ ;

R ⁵ -R ⁷ are the same or different, each selected from hydrogen, halogen, hydroxyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkyl, SR and N(R ¹² ) ₂ ;

Q is C ₁ -C ₆ alkylene, which is unsubstituted or substituted by: (i) unsubstituted or substituted by hydroxy-C ₁ -C ₆ -alkyl, C ₁ -C ₆ -alk Oxygen, phenyl, N(R ¹² ) ₂ and C ₁ -C ₆ alkyl substituted by groups of hydroxy, (ii) hydroxy, with the proviso that said hydroxy is not in each N adjacent to Q of formula (I) At the alpha position of the atom, (iii) CO ₂ R ¹⁰ , or (iv) CON(R ¹² ) ₂ ;

R ⁸ and R ⁹ are the same or different, and each is hydrogen or C ₁ -C ₆ alkyl; or R ⁸ and R ⁹ form a saturated 5 or 6-membered N-containing heterocycle together with the nitrogen atom to which they are attached, and the heterocycle The ring may contain one additional heteroatom selected from O, N, and S; or one of ^R and ^R is an alkylene chain optionally interrupted by O, N, or S, the alkylene chain being separated from The carbon atoms on the alkylene chain represented by Q are connected to form a saturated 5- or 6-membered N-containing heterocyclic ring as defined above;

R ¹⁰ is hydrogen, C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, benzyl or phenyl;

R ¹¹ is C ₁ -C ₆ alkyl, C ₃ -C ₁₀ cycloalkyl, benzyl or phenyl;

Each R ¹² is the same or different and is hydrogen, C ₁ -C ₆ alkyl, cycloalkyl, benzyl or phenyl; or two R ¹² groups together with the nitrogen atom to which they are attached form a saturated 5- or 6-membered N-containing heterocycles which may contain 1 or 2 additional heteroatoms selected from O, N and S; and

n is 1, 2 or 3;

With the proviso that at least one of R ¹ -R ⁴ is other than hydrogen.

In a preferred aspect of the present invention, the benzo[a]phenazinamide-11-derivative has the structure of formula (Ia):

In the formula, R ¹ -R ⁹ are as defined above;

p is 1, 2 or 3; and

R ¹³ is: (i) hydrogen, (ii) C ₁ -C ₆ alkyl unsubstituted or substituted by hydroxyl, aryl or N(R ¹² ) ₂ , wherein R ¹² is as defined above, (iii) CO ₂ R ¹⁰ , (iv) CON(R ¹² ) ₂ or (v) aryl.

When one of R ^{and R} in formula (I) is an alkylene chain connected to a carbon atom on Q, the compound of formula (I) has the following structure (Ib):

In the formula, R ¹ -R ⁷ are as defined in the above formula (I);

R ¹⁴ is hydrogen or C ₁ -C ₆ alkyl;

W is a direct bond or a C ₁ -C ₅ alkylene chain; and

Y and Z together with the N and C atoms to which they are attached form a saturated 5- or 6-membered N-containing heterocycle which may contain an additional O, N or S atom.

C ₁ -C ₆ Alkyl can be linear or branched. C ₁ -C ₆ alkyl is generally C ₁ -C ₄ alkyl, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl or tert-butyl. C ₁ -C ₆ alkyl is unsubstituted or substituted, and is generally replaced by one or more groups selected from hydroxy-C ₁ -C ₆ alkyl (wherein the alkyl moiety is as described herein for C ₁ -C ₆ alkyl Illustratively, substituted by groups of unsubstituted or substituted), C ₁ -C ₆ alkoxy, phenyl, N(R ¹² ) ₂ (wherein R ¹² is as defined above) and hydroxyl. Examples of hydroxy-C ₁ -C ₆ alkyl include, for example, hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl. C ₁ -C ₆ alkylene is a divalent C ₁ -C ₆ alkyl group as defined above.

Aryl is generally an aromatic C ₆ -C ₁₀ carbocyclic group such as phenyl or naphthyl, which is unsubstituted or replaced by halogen, C ₁ -C ₆ alkyl, OH, C ₁ -C ₆ alkoxy, A group substituted by NO ₂ , N(R ¹² ) ₂ , CO ₂ R ¹⁰ , CN or perhalogenated C ₁ -C ₆ alkyl (such as CF ₃ ).

Halogen is F, Cl, Br or I. Preferably it is F, Cl or Br.

C ₁ -C ₆ alkoxy may be linear or branched. Typically C ₁ -C ₄ alkoxy, such as methoxy, ethoxy, propoxy, isopropoxy, n-propoxy, n-butoxy, sec-butoxy or tert-butoxy. C ₁ -C ₆ alkoxy is unsubstituted or substituted, generally replaced by one or more selected from N(R ¹² ) ₂ , CON(R ¹² ) ₂ , hydroxyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, cyano, CO ₂ R ¹⁰ , COR ¹⁰ , saturated 5 or 6 membered N-containing heterocycle or phenyl group , the phenyl group is unsubstituted or substituted by one or more halogen atoms.

C ₃ -C ₁₀ cycloalkyl can be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Usually C ₃ -C ₆ cycloalkyl. C ₂ -C ₆ alkenyl contains one or more unsaturated bonds. The C ₂ -C ₆ alkenyl group may be, for example, vinyl, propenyl, butenyl or pentenyl. C ₂ -C ₆ alkynyl may be ethynyl, propynyl, butynyl or pentynyl. The saturated 5- or 6-membered N-containing heterocycle can be, for example, piperidine, piperazine, morpholine or pyrrolidine.

Heteroaryloxy is an -OHet group where Het is monocyclic or bicyclic and includes unsaturated 5 or 6 membered N-containing heterocycles which may contain one or more additional O, N or S atoms. Examples include furan, thiophene, pyrrole, indole, isoindole, pyrazole, imidazole, isoxazole, oxazole, thiazole, isothiazole, pyridine, quinoline, quinoxaline, isoquinoline, thienopyrazine, Pyran, pyrimidine, pyridazine, pyrazine, puroline and triazine. The aforementioned heterocycles may be unsubstituted or substituted by one or more substituents, for example, by one or more selected from OH, halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, nitro and Substituted by a substituent of amino N(R ¹² ) ₂ as defined above, wherein said C ₁ -C ₆ alkyl group is unsubstituted or substituted, such as substituted by halogen (eg CF ₃ ).

In the compound of the present invention, R ¹ -R ³ in formula (I), (Ia) or (Ib) are each hydrogen, and R ⁴ is not hydrogen. Typically R ⁴ is C ₁ -C ₆ alkoxy, hydroxy, C ₁ -C ₆ alkyl, hydroxy-C ₁ -C ₆ alkyl, nitrile or halogen.

In a series of preferred compounds, R ⁴ in formula (I), (Ia) or (Ib) is C ₁ -C ₆ alkoxy or hydroxyl, R ⁷ is hydroxyl, R ¹ -R ³ , R ⁵ and R ⁶ are each hydrogen. Also preferred are those wherein R ⁴ is C ₁ -C ₆ alkoxy or hydroxy, R ⁶ is C ₁ -C ₆ alkoxy, halogen or methylthio, and R ¹ -R ³ , R ⁵ and R ⁷ are all Hydrogen compounds.

In formula (Ia), R ¹³ is preferably C ₁ -C ₆ alkyl, more preferably methyl.

In formulas (I) and (Ia), Q is preferably a C ₂ or C ₃ alkylene chain, substituted by a C ₁ -C ₆ alkyl group at the alpha position adjacent to the amide nitrogen atom, wherein the C ₁ -C ₆ alkyl is as described above, unsubstituted or substituted. Preferred substituents on Q are unsubstituted C ₁ -C ₆ alkyl or hydroxy-C ₁ -C ₆ alkyl, such as hydroxymethyl or hydroxyethyl. Typically the _C2 or _C3 alkylene chain is substituted at the alpha position adjacent to the amide nitrogen atom by methyl, ethyl, isopropyl, hydroxymethyl, substituted hydroxymethyl or 1-hydroxyethyl .

Examples of preferred compounds of the invention are: Compound name Compound number 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 3 4-Hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide hydrobromide 4 4-Nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 7 9-Bromo-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 10 4-Cyanomethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 11 4-Benzyloxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 12 4-prop-2-ynyloxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 13 4-Ethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 15 4-Isobutoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 16 4-(4-Chloro-benzyloxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 17 4-(2-Methoxy-ethoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 18 4-(2-Hydroxy-ethoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide twenty one 4-(Pyrimidin-2-yloxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide twenty two 4-(2-Morpholin-4-yl-ethoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide twenty three 4-(3-Cyano-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide twenty four 4-Methyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 25 4-Fluoro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 26 4-(3-Dimethylamino-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 27 4-Methylthio (sulfanyl)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 28 4-Carbamoylmethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 29 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (3-amino-2-hydroxy-propyl)-amide 30 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (3-dimethylamino-propyl)-amide 31 4-Bromo-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 32 11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yl acetate 33 4-(2-Oxo-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 34 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide 35 4-Cyano-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 36 Ethyl-carbamic acid 11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yl ester 37 4-Methanesulfonyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 39 4-Chloro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 40 4-Azido-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 41 4-Amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 42 [11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yloxy]acetic acid trifluoroacetate 43 4-Acetylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 44 4-Bis(methylsulfonyl)amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 46 4-(N-Hydroxycarbamimidoyl)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 48 4-Hydroxymethyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 49 11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylic acid trifluoroacetate 50 4-Methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide trifluoroacetate 51 4-Dimethylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 54 4-Methanesulfonylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 55 4-Dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 57 4-(Cyanomethyl-amino)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 59 4,10-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 60 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-propyl)-amide 61 Benzo[a]phenazine-4,11-dicarboxylic acid 4-amide 11-[(2-dimethylamine-ethyl)-amide]trifluoroacetate 62 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1,1-dimethyl-ethyl)-amide 65 4-Methoxy-8-methyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 67 4,10-Dihydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 68 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-propyl)-amide 71 4,10-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide 72 9-Chloro-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 73 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-methyl-propyl)-amide 74 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-hydroxymethyl-ethyl)-amide 75 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-phenyl-ethyl)-amide 76 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-(S)-methyl-ethyl)-amide 77 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-(R)-methyl-ethyl)-amide 78 4-Nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide 79 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-(S)-hydroxymethyl-ethyl)-amide 81 4-Methoxy-10-methylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 82 10-Hydroxy-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(R)-methyl-ethyl)-amide 83 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-hydroxy-propyl)-amide 84 10-Hydroxy-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 85 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-piperidin-1-yl-ethyl)-amide 86 4-Methoxy-benzo[a]phenazine-11-carboxylic acid [1-dimethylamino-1-(2-hydroxyethyl)]-ethylamide 87 10-Amino-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 88 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-morpholin-4-yl-ethyl)-amide 89 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-pyrrolidin-1-yl-ethyl)-amide 90 4-Methoxy-benzo[a]phenazine-11-carboxylic acid {2-[bis(2-hydroxy-ethyl)-amino]-ethyl}-amide 91 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-diethylamino-ethyl)-amide 92 4-Methoxy-9-methylthio-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 93 4,9-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide 94 4,10-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(S)-hydroxymethyl-ethyl)-amide 95 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-methylamino-ethyl)-amide 96 10-Hydroxy-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(S)-hydroxymethyl-ethyl)-amide 97 (R)-4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-methyl-propyl)-amide 98 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (1-methyl-pyrrolidin-3-(R)-yl)-amide 99 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2,3-(bis)-dimethylamino-propyl)-amide 100

Compounds of formula (I) can be prepared by the following methods, which include:

In organic solvents or without solvents, at high temperatures

(a) using an amine of formula (III):

Wherein Q, R and ^R are as defined above, an activated derivative of a compound of formula (II) is ^treated :

In the formula, ^R1 to ^R7 are as defined above; or

(b) treating a compound of formula (IV) with a compound of formula (III) as defined above:

wherein ^R1 to ^R7 and ^R11 are as defined above; and

(c) if desired, converting a resulting benzo[a]phenazine-11-amide derivative of formula (I) into another such derivative, and/or converting a benzo[a]phenazine-11-amide derivative of formula (I) [a] Conversion of phenazine-11-amide derivatives to pharmaceutically acceptable salts thereof.

The optical purity of the resulting compounds with optically active centers (such as benzo[a]phenazine-11-amide derivatives of formula (Ia) and salts thereof) can be determined by adding NMR Shift reagent (such as 2,2,2-trifluoro-1 (9-anthracenyl) ethanol) to determine.

The starting compounds of formula (II) and their esters (compounds of formula (IV)) are novel and thus form a further aspect of the invention.

In step (a), the carboxylic acid group of formula (II) can be activated to the corresponding acid chloride, which can be obtained by treating the free carboxylic acid of formula (II) with thionyl chloride. Alternatively the carboxylic acid group can be activated by treatment with an appropriate amide coupling reagent such as 1,1'-carbonyldiimidazole.

The reaction between the activated derivative of the compound of formula (II) and the amine of formula (III) is generally carried out in an organic solvent. Suitable solvents include dimethylformamide and dichloromethane. The steps of activating the compound of formula (II) and treating the resulting activated derivative with an amine of formula (III) can be carried out without intermediate isolation of the activated derivative. In this case, the process generally involves combining the activator or coupling agent with the compound of formula (II) in an organic solvent and adding the amine of formula (III) to the resulting reaction mixture.

Compounds of formula (II) can be prepared by a process comprising the following steps:

(a) in an organic solvent, optionally in the presence of an acid, with benzoic acid of formula (VI):

or an ester or salt thereof, wherein R ⁵ , R ⁶ and R ⁷ are as defined in formula (I), treating 1,2-naphthoquinone of formula (V):

In the formula, R ¹ to R ⁴ are as defined in formula (I). The solvent may be, for example, ethanol or acetic acid. The regioselectivity of the reaction can be controlled by using 1 to 5 equivalents of mineral acid in the reaction mixture. Use of about 2 equivalents or more of acid, such as 1.5 to 5 equivalents of acid, produces only the desired regioisomer, ie, benzo[a]phenazine-11-carboxylic acid of formula (II). Preferably the inorganic acid is hydrochloric acid, more preferably concentrated hydrochloric acid. The salt of benzoic acid of formula (VI) is typically acetate.

The 1,2-naphthoquinones of formula (V) can be prepared by treating the corresponding 1-tetralones of formula (VII) with selenium dioxide according to the method described in Tetrahedron Letters 1997, 4219-4220,

In the formula, R ¹ to R ⁴ are as defined in formula (I). The 1-tetralone of formula (VII) is a known compound or can be prepared by a known compound by a disclosed method, for example, it can be prepared by the method described in the following reference examples, where necessary, using conventional Laboratory techniques were modified to obtain the desired definitions of ^R1 to ^R4 . Published methods are included in J.Med.Chem 1997, 40 , 3014-3024; J.Org.Chem.1984, 4226; JACS.1994, 116 , pp. 4852-4857 and J.Med.Chem. methods described.

Benzoic acids of formula (VI) are known compounds or can be prepared from known compounds by published methods, with modifications where necessary using routine laboratory techniques to obtain the desired definitions of R ^{to R} . Published methods include those described in J. Chem. Soc. Perkin Trans. I, 1984, p. 2019 and J. Med. Chem 1987, p. 843.

Compounds of formula (II) can also be prepared by a process comprising the following steps:

(a) using naphthylamines of formula (IX):

wherein R ^to R ^are as defined in formula (I), treating 2-halo-3-nitrobenzoic acids of formula (VIII):

In the formula, Hal is Cl, Br, I or F; and

(b) subjecting the resulting compound of formula (X) to reductive cyclization:

In the formula, R ¹ to R ⁴ are as defined above.

Step (a) is generally carried out in an organic solvent. Suitable examples include 2,3-butanediol and ethylene glycol. Step (b) is typically carried out by treating a compound of formula (X) with _NaBH4 in sodium methoxide, sodium ethoxide or aqueous NaOH. This method is described in J. Med. Chem 1987, 30 , pp. 843-851.

Compounds of formula (IV) can be prepared by esterifying the corresponding compound of formula (II) under standard reaction conditions, for example by treating the free compound of formula (II) with an alcohol of formula R ¹¹ -OH (wherein R ¹¹ is as defined above). Carboxylic acid compounds are prepared.

Amines of formula (III) are known and commercially available compounds, or may be prepared from commercial starting materials using conventional techniques, such as those described in Reference Example 2 below.

A compound of formula (I) can be converted into another compound of formula (I) by conventional methods. For example, a compound of formula (I) containing an esterified hydroxyl group such as -OCOMe can be converted to a compound of formula (I) containing a free hydroxyl group by hydrolysis, such as basic hydrolysis. Compounds of formula (I) containing free hydroxyl groups can be converted into compounds of formula (I) containing esterified hydroxyl groups by esterification, eg reaction with a suitable carboxylic acid, acid halide or anhydride. Compounds containing free hydroxyl groups can also be converted into compounds containing carbamate groups eg by treatment with triethylamine and ethyl isocyanate in an aprotic polar solvent such as dimethylformamide.

A compound of formula (I) containing a nitro group can be converted into a compound of formula (I) containing an amino group by reduction, eg, treatment with indium and saturated NH ₄ Cl solution in an organic solvent.

Compounds containing C ₁ -C ₆ alkoxy groups can be converted, for example, by treatment with boron tribromide in halogenated hydrocarbon solvents such as dichloromethane or sodium thioethoxide in dimethylformamide to hydroxy-containing compounds. compound of. Compounds containing hydroxy groups can be converted into compounds containing optionally substituted C ₁ -C ₆ alkoxy groups, for example by treatment with a suitable alkylating agent in the presence of a base. Compounds containing carboxyl groups can be converted to compounds containing hydroxymethyl groups by reduction, for example, by treatment with _LiAlH4 in THF.

A halogen-containing compound can be converted into an alkylthio- or alkoxy-containing compound, for example, by treatment with a solution of a thioalkoxide or alkoxide, respectively, in an organic solvent. Compounds containing a nitrile group can be converted into compounds containing N-hydroxycarbamimidoyl, for example by treatment with hydroxylamine (optionally in the form of a salt) in the presence of a base such as potassium carbonate.

Compounds substituted with an alkylaminomethyl group at a phenyl ring position can be treated under Mannich reaction conditions by treating a compound substituted with a hydroxyl group adjacent to the (unsubstituted) ring position in question with acetic acid, followed by alkylamine and formaldehyde aqueous solution to prepare. Compounds of formula (I) may be acetylated, for example, by treatment with acetyl chloride under suitable conditions to form the acetamido substituent.

The benzo[a]phenazine-11-amide derivative can be converted into a pharmaceutically acceptable salt by a conventional method, and the salt can also be converted into the free compound. The pharmaceutically acceptable salts of the benzo[a]phenazine-11-amide derivatives of formula (I) include inorganic acid salts, such as salts of hydrochloric acid, hydrobromic acid and sulfuric acid; and organic acid salts, such as acetic acid , oxalic, malic, methanesulfonic, trifluoroacetic, benzoic, citric and tartaric acids. When any one of R ¹ -R ⁴ and R ¹⁰ in the compound of formula I is COOH, the salts include the above salts and sodium salts prepared by treating the compound of formula I or its acid salt with the corresponding metal base or ammonia , potassium, calcium and ammonium salts.

Multidrug resistance (MDR) refers to the phenomenon in which cells normally sensitive to chemotherapeutic agents develop resistance to these therapeutic agents as well as to a wide variety of unrelated agents. MDR represents a major obstacle in the successful clinical treatment of cancer. Cancer cells exhibiting MDR can display a number of different cellular alterations, including overexpression of P-glycoprotein (P-gp), overexpression of multidrug resistance-associated protein (MRP), and decreased levels of topoisomerase II (known as atypical drug resistance) and a qualitative change in the expression of topoisomerase I. MDR is a very serious clinical problem, and many tumors develop resistance to many chemotherapeutic agents, including those that specifically target topoisomerase I and/or topoisomerase II.

By simultaneously inhibiting topoisomerases I and II, some compounds such as DACA (Finlay et al., Eur. J. Cancer 32A, 708-714, 1996) have shown Resistance to camptothecin or amsacrine does not result in loss of activity. Distinct cell cycle events have been obtained with inhibitors of topoisomerase I or II (Kaufman, Biochim. Biophys. Acta 1400, 195-212, 1998). Combination inhibitors of topoisomerases I and II appear to combine the properties of individual specific inhibitors and act throughout the cell cycle (Haldane et al., Cancer Chemother. Pharmacol. 32:463-470, 1993), resulting in It has great antitumor activity (Riou et al., Cancer Res. 53, 5987-5993, 1993).

Known due to membrane transport proteins such as P-glycoprotein (Gottesman et al., Annu.Rev.Biochem.62, 385-427, 1993) and MRP (Loe et al., Eur.J.Cancer 32A, 945-957, 1996) The MDR produced by the overexpression of β-β will reduce the clinical effect of chemotherapeutic agents such as paclitaxel, etoposide and doxorubicin. Agents that avoid this MDR mechanism are therefore expected to show therapeutic benefit in cancer treatment.

In biological experiments, it was found that the benzo[a]phenazine-11-amide derivatives of formula I, their pharmaceutically acceptable salts and hydrates and their solvates (hereinafter referred to as "compounds of the present invention") have as topology Activity of inhibitors of isomerases I and II. In one aspect of the invention, the compounds of the invention are combined inhibitors of topoisomerase I and topoisomerase II.

The compounds of the invention are therefore useful as inhibitors of topoisomerase I, or the compounds of the invention are useful as inhibitors of topoisomerase II. In another embodiment they are useful as combined inhibitors of topoisomerase I and topoisomerase II. They have been shown to kill tumor cells in humans and avoid the MDR mechanism. Therefore, they have the potential to treat cancer. Cancer types treatable with the compounds of the invention include leukemias, lymphomas, sarcomas, carcinomas and adenocarcinomas. Specific examples include breast, colon, brain, lung, ovarian, pancreas, stomach and skin cancers.

Human or animal patients harboring tumors can be treated by a method comprising administering thereto a compound of the invention. In particular, a method of treating human tumors, including tumors exhibiting MDR, such as the types of MDR described above, comprises administering to a patient with latent tumor a therapeutically effective amount of a compound of the invention. Various types of tumors can thus be treated, including those that exhibit MDR and those that do not. The compounds of the present invention are administered in an amount effective to reduce or eliminate tumors. In one aspect of the invention, the compounds of the invention are administered orally. In another aspect of the invention, the compounds of the invention are administered parenterally, such as intravenously.

Due to their activity as inhibitors of topoisomerase I and topoisomerase II, the compounds of the invention are also useful as antiviral, antibacterial or antifungal agents.

The compounds of the invention can be administered in various dosage forms, for example orally, eg in the form of tablets, capsules, sugar-coated or film-coated tablets, liquid solutions or suspensions, or parenterally, eg intramuscularly, intravenously or subcutaneously. medication. The compounds of the invention may thus be administered by injection or infusion.

The dosage to be administered depends on various factors including the age, weight and symptoms of the patient and the route of administration. However, generally, when the compound of the present invention is administered alone to an adult, the dosage used for each route of administration is 0.001-500 mg, most commonly 0.01-100 mg per kg body weight. For example, the dose may be administered 1-5 times daily by bolus infusion, infusion over several hours and/or repeated administration.

The benzo[a]phenazine-11-amide derivative of formula (I) or a pharmaceutically acceptable salt thereof can be formulated into a pharmaceutical or veterinary combination comprising a pharmaceutically or veterinarily acceptable carrier or diluent things. The composition is generally prepared according to a usual method, and administered in a pharmaceutically or veterinary appropriate manner. There is thus provided a medicament comprising a compound of the invention for use in the treatment of tumors, including those tumors exhibiting MDR.

The compounds of the invention can be administered in any of the usual ways:

A) Orally, as tablets, coated tablets, lozenges, troches, lozenges, aqueous or oily suspensions, liquid solutions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions for oral administration can be prepared according to any well-known method for preparing pharmaceutical compositions in the art, and such compositions can contain one or more ingredients selected from sweeteners, flavoring agents, coloring agents and preservatives. Reagents to provide pharmaceutically aesthetic and palatable formulations.

Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, dextrose, sucrose, cellulose, corn starch, potato starch, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as cornstarch, alginic acid, alginate or sodium starch glycolate; binders such as starch, gelatin or acacia; lubricants such as silicon dioxide, magnesium or calcium stearate, stearic acid or Talc; boiling complexing agents; coloring agents, sweeteners, humectants such as lecithin, polysorbate or lauryl sulfate. Tablets may be uncoated or may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing delayed action over a longer period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. These preparations can be prepared by known methods, for example by mixing, granulating, tabletting, sugar-coating or film-coating.

Oral formulations may also be hard gelatin capsules, in which the active ingredient is admixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or soft gelatin capsules, in which the active ingredient is present alone or in combination with an aqueous or oily medium such as peanut oil, liquid paraffin or olive oil) in combination.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. These excipients are suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth and acacia; dispersing agents or wetting agents The agent may be a naturally occurring phospholipid such as lecithin, or the condensation product of an alkylene oxide with a fatty acid such as polyoxyethylene stearate, or a condensation product of ethylene oxide with a long chain fatty alcohol such as heptadecylethylene oxide. Heptadecaethyleneoxy-cetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or ethylene oxide with partial esters derived from fatty acids and hexitols Condensation products of partial esters of sugar-alcohol anhydrides such as polyoxyethylene sorbitan monooleate.

The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, and one or more coloring agents, such as sucrose or saccharin.

Oily suspensions may be prepared by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol.

To provide a palatable oral preparation, sweetening and flavoring agents as mentioned above may be added. These compositions can be preserved by the addition of antioxidants such as ascorbic acid. Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents have been mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as liquid paraffin, or a mixture of these oils. Suitable emulsifiers may be naturally occurring gums such as acacia or tragacanth, naturally occurring phospholipids such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan mono Oleate, and the condensation product of this partial ester with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Syrups, especially for diabetics, may contain substances which serve only as a carrier, such as sorbitol, which are not metabolized to glucose or only in a very small amount.

These preparations may also contain demulcents, preservatives, flavoring and coloring agents;

B) Parenteral administration in the form of sterile injectable aqueous or oleaginous suspensions by subcutaneous, intravenous, intramuscular, intrasternal or infusion techniques. This suspension may be formulated according to the known art using those wetting and suspending agents mentioned above having suitable dispersing effects. The sterile injectable preparation may be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol.

Among the acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition fatty acids such as oleic acid find use in the preparation of injectables.

C) administration by inhalation, in the form of an aerosol or solution for nebulizers;

D) Rectal administration in the form of suppositories prepared by mixing the drug with suitable non-irritating excipients which are solid at normal temperature but liquid at rectal temperature and therefore Melts in the rectum to release the drug. These substances are cocoa butter and polyethylene glycols;

E) Topical administration, in the form of creams, ointments, gels, eye washes, solutions or suspensions.

The daily dosage may vary within wide ranges and will be adjusted to the individual requirements in each particular case. Generally, for adult administration, a suitable daily dose ranges from about 5 mg to about 500 mg, although this upper limit may be exceeded in exigencies. The daily dose can be administered in single dose or in divided doses.

The invention is further illustrated by the following examples.

Reference example 1: the preparation of the compound of general formula (II)

Reference Example 1A. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (II.1)

5-methoxy-[1,2]naphthoquinone (prepared by treating 5-methoxytetralone with selenium dioxide, A. Bekaert et al., Tetrahedron Letters 38, 24, 4219-4220, 1997) (1.98g), 2,3-diamino-benzoic acid diacetate (J.Chem.Soc.Perkin.Trans I, 1984, p. 2019) (4.03g) and concentrated hydrochloric acid (2.2mL) in ethanol ( 20 mL) was heated to reflux for 4 hours. The reaction mixture was cooled and the precipitate was collected by filtration and washed with ethanol and diethyl ether to give the title compound (2.74 g) as a light brown solid.

NMR: (DMSO) 4.05 (s, 3H), 7.50 (1H, d), 7.84-7.87 (1H, m), 7.99 (1H, d), 8.08-8.10 (1H, m), 8.41-8.49 (3H, m), 8.63 (1H,d).

Reference Example 1B.4-Methyl-benzo[a]phenazine-11-carboxylic acid (II.2)

5-Methyl-1-tetralone was prepared from o-tolualdehyde according to the literature (J. Med Chem 1997, 40, 3014-3024). Treatment of 5-methyl-1-tetralone with selenium dioxide as described in Reference Example 1A gave 5-methyl-[1,2]naphthoquinone. The resulting product was reacted with 2,3-diamino-benzoic acid diacetate as described in Reference Example 1A to give the title compound.

NMR.d6-DMSO 9.02 (1H, d), 8.51 (1H, dd), 8.47-8.44 (2H, m), 8.11-8.05 (2H, m), 7.85-7.78 (2H, m), 2.79 (3H, s).

Starting from the appropriate substituted aldehyde, the following compounds of formula (II) are prepared in an analogous manner:

Preparation of 4-fluoro-benzo[a]phenazine-11-carboxylic acid (II.3) from 2-fluorobenzaldehyde, NMR, d6-DMSO 8.97 (1H, d), 8.51 (1H, dd), 8.43 (1H , dd), 8.38(1H, d), 8.11-8.06(2H, m), 7.95(1H, m), 7.80(1H, m);

Preparation of 3,4-dimethoxy-benzo[a]phenazine-11-carboxylic acid (II.4) from 2,3-dimethoxybenzaldehyde;

NMR, CDCl3, 8.79 (1H, d), 8.48 (2H, d), 8.37 (1H, d), 8.05 (1H, t), 7.95 (1H, d), 7.76 (1H, d), 4.04 (3H, s), 3.98 (3H, s).

Reference Example 1C.4-Bromo-benzo[a]phenazine-11-carboxylic acid (II.5)

2-Bromobenzyl bromide was converted to 5-bromotetralone according to the literature (J. Org. Chem. 1984, p. 4226). Treatment of 5-bromotetralone with selenium dioxide gave 5-bromo-[1,2]naphthoquinone as described in Reference Example 1A. The resulting product was coupled with 2,3-diamino-benzoic acid diacetate as described in Reference Example 1A to afford the title compound.

NMR d6-DMSO, 9.24(1H,d), 8.48(2H,m), 8.33(1H,m), 8.26(1H,m), 8.20(1H,d), 8.08(1H,t), 7.87(1H , t).

Reference Example 1D. 4-cyano-benzo[a]phenazine-11-carboxylic acid (II.6)

To a solution of 5-bromotetralone (see Reference Example 1C) (15.0 g) in N,N-dimethylformamide (20 mL) was added copper(I) cyanide (6.39 g) and reacted The mixture was heated to reflux for 20 hours. The reaction mixture was then cooled to 80° C. and a solution of FeCl ₃ .6H ₂ O (24 g) in water (38 mL) was added. After stirring for a further 45 minutes, the resulting reaction mixture was cooled, diluted with water, extracted into toluene, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give 5-cyano-1-tetralone as a yellow solid.

As described in Reference Example 1A, treatment of 5-cyano-1-tetralone with selenium dioxide gave the corresponding 5-cyano-[1,2]naphthoquinone, which was combined with 2,3-dihydronaphthalenone Amino-benzoic acid diacetate coupling affords the title compound.

NMR (d6-DMSO), 8.10-8.18 (2H, m), 8.33 (1H, d), 8.43 (2H, m), 8.48-8.54 (2H, m), 9.48 (1H, d).

MS DCI/NH3 m/z 300 (MH+).

Reference Example 1E.4-Chloro-benzo[a]phenazine-11-carboxylic acid (II.7)

5-Amino-1-tetralone was prepared from α-tetralone according to the literature (J. Am. Chem. Soc. 1994, p. 4852). A mixture of 5-amino-1-tetralone (80 mg) and concentrated hydrochloric acid (1 mL) was cooled to 0°C. A solution of sodium nitrite (35 mg) in water (0.5 mL) was added dropwise to the stirring solution. The cold diazonium salt solution was then quickly poured into a stirred solution of copper(I) chloride (62 mg) in concentrated hydrochloric acid (1 mL). The reaction mixture was allowed to warm to ambient temperature, then stirred for 1.5 hours. The mixture was then extracted with ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give 5-chloro-1-tetralone as a brown solid (87 mg).

As described in Reference Example 1A, 5-chloro-tetralone was treated with selenium dioxide to obtain 5-chloro-[1,2]naphthoquinone, and as described in Reference Example 1A, the resulting product was combined with 2, Coupling of 3-diamino-benzoic acid diacetate affords the title compound.

Reference Example 1F. 4-Methanesulfonyl-benzo[a]phenazine-11-carboxylic acid (II.8)

5-Methylthio-1-tetralone was prepared from 5-hydroxy-1-tetralone according to the literature (J. Med. Chem. 1997, p. 1049). A mixture of 5-methylthio-1-tetralone (221 mg) and 3-chloroperbenzoic acid (595 mg) was stirred at room temperature for 2 hours. The reaction mixture was then extracted with dichloromethane, washed with water, dried ( _Na2SO4 ) and the solvent removed in vacuo to give 5-methanesulfonyl-1-tetralone as an off-white solid (210 mg ₎ .

5-Methanesulfonyl-1-tetralone was treated with selenium dioxide as described in Reference Example 1A to give 5-Methanesulfonyl-[1,2]naphthoquinone, as described in Reference Example 1A, Coupling of the resulting product with 2,3-diamino-benzoic acid diacetate gave the title compound.

NMR, d6-DMSO, 9.61 (1H, d), 9.03 (1H, d), 8.56 (2H, m), 8.43 (1H, d), 8.32 (1H, m), 8.16 (2H, m), 3.51 ( 3H, s). MS DCI/NH3 m/z 353 (MH+).

Reference Example 1G.4-Azido-benzo[a]phenazine-11-carboxylic acid (II.9)

5-Amino-1-tetralone was prepared from α-tetralone according to the literature (J. Am. Chem. Soc. 1994, p. 4852). To a cold solution of 5-amino-1-tetralone (415 mg) in water (2 mL) and concentrated hydrochloric acid (5 mL) was added sodium nitrite (186 mg) in water (1.2 mL), keeping at low temperature. solution. After 40 minutes, a solution of sodium azide (184 mg) in water (1.2 mL) was added dropwise. The reaction mixture was allowed to warm to room temperature. After a further 1 hour the reaction was quenched with water, extracted into diethyl ether, washed with sodium bicarbonate solution, dried (MgSO ₄ ) and the solvent removed in vacuo to give crude product which was purified using flash chromatography to give 5-azido- 1-Tetralone (77 mg).

Treatment of 5-azido-1-tetralone with selenium dioxide as described in Reference Example 1A gave 5-azido-[1,2]naphthoquinone, as described in Reference Example 1A, Coupling of the resulting product with 2,3-diamino-benzoic acid diacetate gave the title compound.

Reference Example 1H. Benzo[a]phenazine-4,11-dicarboxylic acid 4-methyl ester (II.10)

According to literature (J.Org.Chem.1976, page 2918) by 2-methyl-2,6,7,8-tetrahydro-chromen-5-one (Tetrahedron Letters, 1975, page 3407) Preparation of methyl 5-oxo-5,6,7,8-tetrahydro-naphthoate. Treatment of 5-oxo-5,6,7,8-tetrahydro-naphthoic acid methyl ester with selenium dioxide as described in Reference Example 1A gave the corresponding 1,2-naphthoquinone, 5,6-dioxo Substitution of methyl 5,6-dihydro-naphthalene-1-carboxylate and coupling of the resulting product with 2,3-diamino-benzoic acid diacetate as described in Reference Example 1A gave the title compound.

NMR, d6-DMSO, 9.47 (1H, d), 9.02 (1H, d), 8.53 (1H, d), 8.44 (2H, m), 8.20 (1H, d), 8.10 (2H, m), 4.03 ( 3H, s).

Reference Example 1I.4-Ethoxy-benzo[a]phenazine-11-carboxylic acid (II.11)

A mixture of 5-hydroxy-1-tetralone (2.00 g) and sodium hydroxide (493 mg) in ethanol (40 mL) was warmed to 50°C. Ethyl iodide (3.94 mL) was then added and the resulting reaction mixture was heated to reflux for 16 hours. The reaction mixture was diluted with 2N hydrochloric acid, extracted into ethyl acetate, dried and the solvent removed in vacuo to give 5-ethoxy-1-tetralone as a white solid (2.06g).

As described in Reference Example 1A, treatment of 5-ethoxy-1-tetralone with selenium dioxide gave 5-ethoxy-[1,2]-naphthoquinone, which was combined with 2 , 3-Diamino-benzoic acid diacetate coupling affords the title compound.

NMR: d6-DMSO.1.52 (3H, q), 4.33 (2H, t), 7.50 (1H, d), 7.85-7.90 (1H, m), 8.00 (1H, d), 8.08-8.11 (1H, m ), 8.45-8.55 (3H, m), 8.68 (1H, d).

Reference Example 1J.4-Methylthio-benzo[a]phenazine-11-carboxylic acid (II.12)

To a stirred solution of 4-fluoro-benzo[a]phenazine-11-carboxylic acid (II.3, see Reference Example 1B) (58 mg) in dry DMSO (3 mL) was added sodium thiomethoxide ( 55 mg) and the resulting reaction mixture was heated at 100°C for 1.5 hours and at 130°C for 1 hour. The reaction mixture was then cooled to room temperature, quenched with acetic acid, extracted with ethyl acetate, washed with water, dried and the solvent removed in vacuo to afford the title compound as a red solid (37mg).

NMR, d6-DMSO, 9.00 (1H, d), 8.52 (2H, m), 8.45 (1H, d), 8.11 (2H, m), 7.91 (2H, m), 2.70 (3H, s).

MS, DCI/NH3m/z=321 (MH+, 100%).

Reference Example 1K.4-Benzyloxy-benzo[a]phenazine-11-carboxylic acid (II.13)

To a solution of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1, see Reference Example 1A) (441 mg) in dichloromethane (30 mL) cooled to 0° C. M A solution of boron tribromide in dichloromethane (7.25 mL). The reaction mixture was warmed to room temperature and stirred for 16 hours. The mixture was then poured into ice/water to give 4-hydroxy-benzo[a]phenazine-11-carboxylic acid as a red/brown solid (230 mg), which was collected by filtration and air-dried.

NMR: d6-DMSO.7.35(1H,d), 7.75(1H,m), 7.92(1H,d), 8.08(1H,m), 8.47-8.55(4H,m), 10.72(1H, broad peak) .

A mixture of 4-hydroxy-benzo[a]phenazine-11-carboxylic acid (80 mg), sodium hydroxide (34 mg) and benzyl bromide (100 μL) in ethanol (2 mL) was heated to reflux for 4 hours. The reaction mixture was then cooled, diluted with ethyl acetate, washed with dilute acid, dried ( _MgSO4 ) and the solvent removed in vacuo to afford the crude title compound as a brown solid (50 mg).

NMR: d6-DMSO. includes 5.35 (2H, s).

The following compounds were prepared in a similar manner from 4-hydroxy-benzo[a]phenazine-11-carboxylic acid using the appropriate alkylating reagent:

Preparation of 4-prop-2-ynyloxy-benzo[a]phenazine-11-carboxylic acid (II.14) using propargyl bromide;

Preparation of 4-isobutoxy-benzo[a]phenazine-11-carboxylic acid (II.15) using isobutyl bromide;

Preparation of 4-(4-chloro-benzyloxy)-benzo[a]phenazine-11-carboxylic acid (II.16) using p-chlorobenzyl bromide;

Preparation of 4-(2-methoxy-ethoxy)-benzo[a]phenazine-11-carboxylic acid (II.17) using 2-bromoethylmethyl ether;

Preparation of 4-ethoxycarbonylmethoxy-benzo[a]phenazine-11-carboxylic acid (II.18) using ethyl bromoacetate, sodium ethoxide in absolute ethanol was used for the reaction;

4-[2-(tert-Butyl-dimethyl-silanyloxy)-ethoxy]-benzo[a ] phenazine-11-carboxylic acid (II.19). tert-Butyl-(2-iodo-ethoxy)-dimethylsilane was prepared from 2-iodo-ethanol using standard methods.

Reference Example 1L. Benzo[a]phenazine-11-carboxylic acid (II.20)

A mixture of 1,2-naphthoquinone (commercial product, 2.0 g) and 2,3-diamino-benzoic acid diacetate (3.79 g) in acetic acid (30 mL) was heated to reflux for 2 hours. The reaction mixture was cooled and the solvent was removed in vacuo to give a pure gum. The resulting product was purified using flash chromatography to give a 2:1 mixture of the desired title compound and the undesired benzo[a]phenazine-8-carboxylic acid (840 mg). The two isomers were separated after further modification (see Reference Example 3B below).

NMR d6-DMSO, including 9.07-9.09 (1H, m), 9.21-9.22 (1H, m). 2:1 ratio.

Reference Example 1M.4-Methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.21) and 3-methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid ( II.22)

A solution of methyl benzo[a]phenazine-11-carboxylate (IV.20, see Reference Example 3B, 220 mg) in chlorosulfonic acid (2 mL) was heated to 180° C. under nitrogen for 6 hours. The reaction was then cooled, poured into ice/water and the pale yellow solid collected by filtration to yield 4-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid and 3-chlorosulfonyl-benzo[a]phenazine - an approximately 1:1 mixture of 11-carboxylic acid (182 mg). NMR d6-DMSO, including 9.41 (1H, s), 9.30 (1H, d).

A mixture of 4-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid and 3-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid (182 mg) was dissolved in dichloromethane (5 mL) . A 40% aqueous methylamine solution (5 mL) was added thereto, and the reaction mixture was vigorously stirred for 4 hours. The reaction mixture was then poured into dichloromethane, acidified (2N HCl), extracted into dichloromethane, dried ( _MgSO4 ) and the solvent removed in vacuo to give an approximately 1:1 mixture of the title compounds (160mg). The two isomers were separated after further chemical modification.

NMR (CDCl3+d4MeOH), including 9.38 (1H, s), 9.22 (1H, d), 2.61 (3H, s), 2.55 (3H, s).

Prepared by reaction of dimethylamine and a mixture of 4-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid and 3-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid in a similar manner 4-Dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.23) and 3-dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.24 ). The two isomers were separated after further chemical modification. NMR (d6-DMSO) of the mixture included 9.06 (1H, d), 9.42 (1H, s), 2.93 (6H, s).

Prepared by reaction of ammonium hydroxide and a mixture of 4-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid and 3-chlorosulfonyl-benzo[a]phenazine-11-carboxylic acid in a similar manner 4-Sulphamoyl-benzo[a]phenazine-11-carboxylic acid (II.25) and 3-sulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.26). The two isomers were separated after further chemical modification. NMR (d6-DMSO) of the mixture included 9.39 (1H, d), 9.60 (1H, s).

Reference Example 1N. 4-Nitro-benzo[a]phenazine-11-carboxylic acid (II.27) and 3-nitro-benzo[a]phenazine-11-carboxylic acid (II.28)

Concentrated sulfuric acid (5 mL) and concentrated nitric acid (5 mL) were mixed together at 0°C. To the mixture was added benzo[a]phenazine-11-carboxylic acid (II.20) (100 mg), and the resulting mixture was slowly warmed to room temperature. After 24 hours the reaction mixture was poured into water to give a yellow precipitate. The precipitate was collected by filtration to give 4-nitro-benzo[a]phenazine-11-carboxylic acid (II.27) and 3-nitro-benzo[a]phenazine-11-carboxylic acid (II.28) The 4:1 mixture. The two isomers were separated after further chemical modification.

NMR d6-DMSO, including 9.82 (1H, d), 9.46 (1H, d). 1:4 ratio.

MS MH@320.

Reference Example 10.4-Amino-benzo[a]phenazine-11-carboxylic acid (II.29) and 3-nitro-benzo[a]phenazine-11-carboxylic acid (II.30)

To 4-nitro-benzo[a]phenazine-11-carboxylic acid (II.27) and 3-nitro-benzo[a]phenazine-11 in ethanol (5 mL) from Reference Example 1N - To a 4:1 mixture (52 mg) of formic acid (II.28) was added ammonium chloride solution (3 mL) and indium (catalyst). The reaction mixture was heated to reflux, then cooled and filtered through a bed of celite. The filtrate was diluted with water, extracted into dichloromethane, dried ( _MgSO4 ) and the solvent removed in vacuo to give a mixture of the title compounds (48mg). Separated after further modification.

NMR, d6-DMSO, 6.95(1H,d), 7.55(1H,t), 7.75(2H,m), 8.05(2H,m), 8.20(1H,d), 8.65(1H,d).

Reference Example 1P. 3-Bromo-4-hydroxy-benzo[a]phenazine-11-carboxylic acid (II.31)

A mixture of 4-hydroxy-benzo[a]phenazine-11-carboxylic acid (see Reference Example 1K, 102 mg) and bromine (0.04 mL) was stirred in chloroform (3 mL) at room temperature for 20 hours. The solvent was removed in vacuo and the residue was purified using flash chromatography (10% methanol in dichloromethane) to give the title compound as a yellow-brown solid (24 mg).

MS DCI/NH3, MH+369/371 (1:1)

NMR (CDCl3), 7.94 (1H, d), 8.1-8.2 (2H, m), 8.52-8.62 (2H, m), 9.1 (1H, d).

Reference Example 1Q.2-Nitro-benzo[a]phenazine-11-carboxylic acid (II.32)

7-Nitro-1-tetralone was prepared according to the literature (J. Am. Chem. Soc, 1994, 116 , pp. 4852-4857). Treatment of 7-nitro-1-tetralone with selenium dioxide as described in Reference Example 1A gave the corresponding 7-nitro-[1,2]-naphthoquinone as described in Reference Example 1A , coupling the resulting product with 2,3-diamino-benzoic acid diacetate to give the title compound.

Reference Example 1R.2-Methoxy-benzo[a]phenazine-11-carboxylic acid (II.33)

8-Amino-naphthalene-2-ol was prepared from 8-amino-2-naphthalenesulfonic acid (commercial product) according to the literature (J. Org. Chem. 1949, p. 351). To a solution of 8-amino-naphthalene-2-ol (8.00 g) in anhydrous N,N-dimethylformamide (80 mL) was carefully added sodium hydride (60% dispersion in mineral ). After stirring for 4 hours, the reaction mixture was cooled in an ice bath and methyl iodide (3.13 mL) was added dropwise. The reaction mixture was then stirred at room temperature for 3 days. Water (10 mL) was then added and the volatiles were removed in vacuo. The resulting residue was dissolved in chloroform, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a dark oil. The product was purified using flash chromatography (chloroform) to give 7-methoxy-naphthalen-1-ylamine as a dark brown liquid (2.92 g).

Treatment of 7-methoxy-naphthalen-1-ylamine with 2-bromo-3-nitro-benzoic acid gave 2- (7-Methoxy-naphthalen-1-ylamino)-3-nitro-benzoic acid. Reductive cyclization using sodium borohydride (J. Med Chem. 1987, p. 843) affords the desired title compound.

NMR, d6-DMSO, 14.44 (1H, broad peak, s), 8.55 (1H, d), 8.48 (1H, dd), 8.42 (1H, dd), 8.23 (1H, d), 8.10-8.04 (2H, m), 7.86 (1H, d), 7.56 (1H, dd), 4.02 (3H, s).

MH+@305

3-Methoxy-benzo[a]phenazine-11-carboxylic acid (II.34) was prepared in a similar manner starting from 5-amino-naphthalene-2-sulfonic acid (commercial product).

NMR, d6-DMSO, 14.60 (1H, broad peak, s), 8.94 (1H, d), 8.51 (2H, d), 8.23 (1H, d), 8.10-7.96 (2H, m), 7.67 (1H, d), 7.55 (1H, dd), 3.99 (3H, s).

MH+@305

Reference Example 1S. 9-Bromo-4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.35)

A mixture of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) (100 mg) and bromine (5 drops) in chloroform (7 mL) was stirred at room temperature for 3 days. The reaction mixture was reduced in vacuo and the desired product (25 mg) was isolated using flash chromatography.

MS MH+@383/385(1∶1)

NMR, CDCl3, 14.47 (1H, br, s), 8.86 (1H, d), 8.33 (1H, d), 7.95 (1H, t), 7.78 (1H, d), 6.97 (1H, d), 6.12 ( 1H, d), 5.75 (1H, d), 3.94 (3H, s).

Reference Example 1T. 4,10-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (II.36)

2-Amino-6-methoxy-3-nitro-benzoic acid methyl ester was prepared using a method similar to that described in the literature (Kim et al., J. Med. Chem. 1993, p. 2335). Hydrolysis of the methyl ester was achieved by refluxing in ethanol for 2 hours using potassium hydroxide to afford 2-amino-6-methoxy-3-nitro-benzoic acid. Hydrogenation of the nitro group was carried out in acetic acid/water with palladium on carbon as a catalyst under 50 psi of _H2 in a Parr apparatus to give 2,3-diamino-6-methoxy-benzoic acid. The resulting product was reacted with 5-methoxy-[1,2]naphthoquinone as described in Reference Example 1A to give the desired title compound.

NMR, d6-DMSO, 4.05(3H, s), 4.10(3H, s), 7.45(1H, d), 7.78-7.82(1H, m), 7.92(1H, d), 8.03(1H, d), 8.36-8.40 (2H, m), 8.78 (1H, d).

Reference Example 1U.4-Methoxy-8-methyl-benzo[a]phenazine-11-carboxylic acid (II.37)

2,3-Diamino-4-methylbenzoic acid was prepared from 4-methylanthranilic acid according to the method described by Rewcastle et al. (J. Med. Chem. 1987, p. 843). The resulting product was reacted with 5-methoxy-[1,2]naphthoquinone as described in Reference Example 1A to give the desired title compound.

NMR, d6-DMSO, 2.97(3H, s), 4.06(3H, s), 7.05(1H, d), 7.85-7.90(1H, m), 7.97(1H, d), 8.02(1H, d), 8.45-8.50 (2H, m), 8.57 (1H, d).

Reference Example 1V.9-Chloro-4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.38)

5-Chloro-3-nitroanthranilic acid was prepared according to the method described by Flippin et al. (Biorg. Med. Chem. Letts 1996, p. 477). This material was hydrogenated in ethyl acetate using palladium on carbon under 50 psi of _H2 for 2 hours to afford 2,3-diamino-5-chloro-benzoic acid. The resulting product was reacted with 5-methoxy-[1,2]naphthoquinone as described in Reference Example 1A to give the desired title compound.

NMR, d6-DMSO, 4.05(3H,s), 7.48(1H,d), 7.82-7.86(1H,m), 7.92(1H,d), 8.30(1H,d), 8.48(1H,d), 8.54 (1H, d), 8.68 (1H, d), 14.1 (1H, broad peak).

Reference example 2: the preparation of the compound of general formula (III)

Reference Example 2A. 4-Aza-DL-leucine methyl ester hydrochloride (III.1)

Saturate methanol (150 mL) with anhydrous hydrogen chloride gas. To the resulting mixture was added 4-aza-DL-leucine (4.86 g) (commercial product), and the resulting reaction mixture was stirred overnight at room temperature. The solvent was removed in vacuo to afford the title compound (quantitative preparation).

Reference example 2B. N ¹ , N ¹ -dimethyl-butane-1,2-diamine (III.2)

Methyl N-(tert-butoxycarbonyl)-2-aminobutyrate was prepared as described in the literature (J. Med. Chem. 1989, p. 1886). Treatment of this compound with diisobutylaluminum hydride in toluene at -78°C for 1.5 hours affords the corresponding aldehyde (see H.W. Scheeren et al., J. Org. Chem. 1990, p. 3998).

The resulting mixture of aldehyde (1.85 g), dimethylamine hydrochloride (1.61 g), sodium acetate (1.21 g) and sodium cyanoborohydride (0.83 g) in methanol was stirred at room temperature for 24 hours. The pH was adjusted to 6-7 with acetic acid and monitored during the reaction. The reaction mixture was then concentrated in vacuo, the residue was dissolved in ethyl acetate and washed with water. The organic layer was dried ( _MgSO4 ) and the solvent was removed in vacuo to give a colorless oil, the resulting product was purified using flash chromatography to give the desired dimethylamine derivative as a pale oil (0.56g). To this compound (260 mg) was carefully added 4.0 M HCl in dioxane (2 mL), and the reaction mixture was stirred for 90 minutes. The reaction mixture was then concentrated in vacuo to afford the desired title compound as an off-white solid (quantitative preparation).

Reference Example 2C. ³ N ¹ , N ¹ -trimethyl-butane-1,2-diamine hydrochloride (III.3)

N-(tert-butoxycarbonyl)-DL-valine methyl ester was prepared from DL-valine using standard preparative techniques. The resulting product was converted into the desired title compound using a method similar to that in Reference Example 2B. Preparation of enantiomerically pure (R) ^-3N1 , ^N1 -trimethyl-butane-1,2 ^- diamine hydrochloride using D-valine as starting material (III.3.a) .

Reference Example 2D. N ¹ , N ¹ -Dimethyl-3-phenyl-propane-1,2-diamine hydrochloride (III.4)

N-(tert-butoxycarbonyl)-DL-phenylalanine methyl ester was prepared from DL-phenylalanine using standard preparative techniques. The resulting product was converted into the desired title compound using a method similar to that in Reference Example 2B.

Reference example 2E. (S)-N ¹ , N ¹ -dimethyl-propane-1,2-diamine hydrochloride (III.5)

2-(S)-[N-(tert-butoxycarbonyl)amino]propanal was prepared from L-alanine according to the method described in the literature (Chakravarty et al., J. Med. Chem. 1989, p. 1886) . The resulting mixture of aldehyde (2.62 g), dimethylamine hydrochloride (2.47 g), sodium acetate (1.99 g) and sodium cyanoborohydride (1.43 g) in methanol (45 mL) was stirred at room temperature for 18 hours . The resulting reaction mixture was dissolved in ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a viscous oil. The resulting product was dissolved in dichloromethane, extracted with citric acid, basified with sodium hydroxide and extracted again with ethyl acetate. The organic layer was concentrated in vacuo to give the dimethylamine derivative as a white solid (586 mg).

To this compound (366 mg) was added a 4.0 M solution of hydrochloric acid in dioxane (5.5 mL) at room temperature. After stirring for 30 minutes, the volatiles were removed in vacuo to give the desired title compound as a viscous oil (313mg).

Reference example 2F. (R)-N ¹ , N ¹ -dimethyl-propane-1,2-diamine hydrochloride (III.6)

2-(R)-[N-(tert-butoxycarbonyl ) amino] propionaldehyde.

The resulting mixture of aldehyde (16.21 g), dimethylamine hydrochloride (15.28 g), sodium acetate (11.53 g) and sodium cyanoborohydride (8.24 g) in methanol (250 mL) was stirred at room temperature for 18 hours , maintained at a pH of 6-7 using AcOH. The resulting reaction mixture was dissolved in ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a viscous oil. The resulting product was purified by flash chromatography to give the dimethylamino derivative as a white solid (10.81 g).

To this compound (3.17 g) was added a 4.0 M solution of hydrochloric acid in dioxane (20 mL) at room temperature. After stirring for 1 hour, the volatiles were removed in vacuo to give the desired title compound as a viscous oil (2.79g).

Reference Example 2G. (S)-2-amino-3-dimethylamino-propan-1-ol hydrochloride (III.7)

N-[(tert-butoxy)carbonyl]-O-(tert-butoxy)carbonyl]-O-(tert- Butyldimethylsilyl)-R-serine methyl ester. Treatment of this compound with diisobutylaluminum hydride in toluene at -70°C for 2 hours affords the corresponding aldehyde (see H.W. Scheeren et al., J. Org. Chem. 1990, p. 3998).

The resulting mixture of crude aldehyde (4.43 g), dimethylamine hydrochloride (2.26 g), sodium cyanoborohydride (1.31 g) and sodium acetate (1.83 g) in methanol (55 mL) was stirred at room temperature for 24 Hour. Dimethylamine derivatives are obtained after aqueous work-up. The resulting product was dissolved in dioxane, and to the resulting mixture was added a 4.0 M solution of hydrochloric acid in dioxane, and the mixture was stirred for 20 minutes. The mixture was concentrated in vacuo to afford the crude desired title compound as a white solid.

Reference Example 2H.3(S)-amino-4-dimethylamino-butan-2(S)-alcohol hydrochloride (III.8)

Hydrogen chloride gas was bubbled through a solution of D-threonine (20 g) in methanol (50 mL). The resulting solution was stirred at room temperature for 5 hours and concentrated in vacuo to give D-threonine methyl ester hydrochloride as a white solid (prepared quantitatively). The acetonitrile solution of di-tert-butyl dicarbonate was treated with triethylamine to obtain N-(tert-butoxycarbonyl)-D-threonine methyl ester. Treatment of this compound with tert-butyldimethylsilyl chloride in dichloromethane and imidazole affords the corresponding TBDMS-protected alcohol.

Treatment of this compound with diisobutylaluminum hydride in toluene at -78°C for 4 hours affords the corresponding aldehyde (see H.W. Scheeren et al., J. Org. Chem. 1990, p. 3998). Reductive amination was carried out as described in Reference Example 2G to give the corresponding dimethylamino derivative. Deprotection using 4.0 M HCl in dioxane as described in Reference Example 2G afforded the title compound as a golden oil.

Reference example 2I.3-amino-4-dimethylamino-butan-1-ol hydrochloride (III.9)

Treatment of DL-homoserine with hydrogen chloride gas in methanol yields the corresponding lactone. Treatment of the resulting product with di-tert-butyl dicarbonate in acetonitrile and triethylamine gave the N-tert-butoxycarbonyl protected derivative. Treatment of this compound with diisobutylaluminum hydride in toluene at -78°C for 4 hours affords the corresponding lactol (see H.W. Scheeren et al., J. Org. Chem. 1990, p. 3998).

Reductive amination was performed as described in Reference Example 2G to afford the corresponding dimethylamino derivative, 3-[N-(tert-butoxycarbonyl)amino]-4-dimethylamino-butan-1-ol. Deprotection using 4.0 M HCl in dioxane as described in Reference Example 2G afforded the desired title compound.

Reference Example 2J.1-Methyl-3-(R)-aminopyrrolidine hydrochloride (III.10)

A solution of di-tert-butyl dicarbonate (1.06 g) in tert-butanol (5 mL) was added dropwise to treat 3R-(-)-1-benzyl-3-aminopyrrolidine (commercial product, 847 mg) in tert-butyl Alcohol (10 mL) and 1.0 N sodium hydroxide solution (4.8 mL). After 1.5 h the tert-butanol was removed in vacuo and the residue was dissolved in ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give the desired 3N-tert-butoxycarbonyl protected derivative as a colorless Pure gum (1.26g).

A solution of 3N-tert-butoxycarbonyl-protected derivative (800 mg) in tetrahydrofuran was stirred over palladium hydroxide catalyst for 24 hours under hydrogen atmosphere. The reaction mixture was then filtered through celite and the solvent was removed in vacuo to afford the desired 3N-tert-butoxycarbonyl-3-(R)-aminopyrrolidine (quantitatively prepared).

To a solution of N-tert-butoxycarbonyl-3-(R)-aminopyrrolidine (437 mg) in methanol (10 mL) were added 37% aqueous formaldehyde (0.52 mL) and sodium borohydride (271 mg). The resulting reaction mixture was stirred at room temperature for 24 hours, then concentrated in vacuo. The residue was dissolved in chloroform, washed with brine and NaHCO ₃ solution, dried (MgSO ₄ ) and the solvent removed in vacuo to give the desired 3N-tert-butoxycarbonyl-3-(R)-amino-1-methylpyrrole Alkanes pure gum (390 mg). Deprotection using 4.0M HCl in dioxane as described above affords the desired title compound.

Reference Example 2K. N ¹ , N ¹ , N ² , N ² -tetramethyl-propane-1,2,3-triamine trihydrochloride (III.11)

To ice-cooled thionyl chloride (35 mL) was added 1,3-bis(dimethylamino)-propan-2-ol (4.91 g, 33.58 mmol, commercial product) dropwise with stirring over 45 minutes. After completion of the dropwise addition, the resulting mixture was further stirred for 4 hours. Excess thionyl chloride was removed under reduced pressure to give 8.9 g of the product as the hydrochloride salt as a cream solid. The free base was obtained by treating a suspension of the hydrochloride salt in toluene (18 mL) with a solution of sodium hydroxide (2.4 equiv) in water (14 mL) for 30 minutes. The organic layer was removed, dried over _MgSO4 , filtered and the solvent removed in vacuo to give 2-chloro-N,N, ^N1 , ^N1 -tetramethylpropane-1,3-diamine as a yellow liquid.

Treat 2-chloro-N, N, N ¹ , N ¹ -tetramethylpropane-1,3-diamine (1.6 g, 9.71 mmol) with potassium phthalimide (1.98 g, 10.68 mmol). Toluene (14 mL) solution. The stirred mixture was heated at reflux under an inert atmosphere for 18 hours, cooled to ambient temperature and the solvent removed under reduced pressure to give a light brown solid which was recrystallized from diethyl ether to give a light tan solid (1.85g). A solution of the resulting solid (0.953, 3.46 mmol) in ethanol (10 mL) was treated with hydrazine hydrate (0.22 mL, 6.93 mmol), and the mixture was stirred at ambient temperature for 18 hours. The suspension was removed by filtration and the filtrate was acidified with 2 mL of 2M hydrochloric acid and the solvent was removed in vacuo to give the product as a cream solid (795 mg). M.pt 126.5-128°C, MH ⁺ 432.

Reference example 3: the preparation of the compound of general formula (IV)

Reference Example 3A. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.1)

Acetyl chloride (4.6 mL) was added dropwise to a suspension of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1, see Reference Example 1A, 4.9 g) in methanol (50 mL) . The mixture was heated to reflux for 4 hours. Subsequent removal of volatiles in vacuo afforded the title compound as a dark solid (prepared quantitatively).

NMR, d6-DMSO, 8.76 (1H, d), 8.41 (2H, d), 8.26 (1H, d), 8.00 (1H, t), 7.90 (1H, d), 7.78 (1H, t), 7.43 ( 1H,d), 4.08(3H,s), 4.03(3H,s).

Reference Example 3B. Benzo[a]phenazine-11-carboxylic acid methyl ester (IV.2)

A mixture of benzo[a]phenazine-11-carboxylic acid (II.20) and benzo[a]phenazine-8-carboxylic acid (885 mg), prepared as described in Reference Example 1L above, was dissolved in methanol (40 mL) and acetyl chloride (920 μL) and heated to reflux for 90 minutes. The reaction mixture was then cooled slowly and the title compound was collected as a single isomer (377mg) by filtration.

NMR, d6-DMSO, 9.53-9.55 (1H, m), 9.06 (1H, d), 8.57 (1H, d), 8.47 (1H, d), 8.35 (1H, d), 8.10-8.11 (1H, m ), 8.09-8.02 (1H, m), 7.95-8.01 (2H, m), 4.21 (3H, s).

Reference Example 3C.4-Dimethylamino-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.3) and 3-dimethylamino-benzo[a]phenazine-11-carboxylic acid methyl ester ( IV.4)

4-Amino-benzo[a]phenazine-11-carboxylic acid (II.29) and 3-amino-benzo [a] To a 4:1 mixture (48 mg) of phenazine-11-carboxylic acid (II.30) were added methyl iodide (0.5 mL) and diisopropylethylamine (2.0 mL). The reaction mixture was heated to 100°C for 4 hours. The mixture was cooled, diluted with ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give the title mixture as a red solid (29 mg). Purified after further chemical modification.

Reference Example 3D. 10-Fluoro-4-methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.5)

Methyl 2-amino-6-fluoro-3-nitrobenzoate was prepared according to the literature (J. Med. Chem 1993, p. 2335). Hydrogenation over palladium on carbon in methanol gives 2,3-diamino-6-fluoro-benzoic acid methyl ester. Reaction of said compound with a solution of 5-methoxy-[1,2]naphthoquinone in cold ethanol (acidified with concentrated hydrochloric acid) afforded the desired title compound.

NMR, CDCl3, 4.08(3H, s), 4.20(3H, s), 7.20(1H, d), 7.65-7.75(2H, m), 7.92(1H, d), 8.35(1H, dd), 8.53( 1H,d), 8.91(1H,d).

MS m/e 337 (MH+, 100%)

Reference Example 3E. 4-Methoxy-10-methylamino-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.6)

A mixture of methyl 10-fluoro-4-methoxy-benzo[a]phenazine-11-carboxylate (IV.5) (100 mg) and 2.0 M methylamine in tetrahydrofuran was stirred at room temperature for 18 hours. Subsequent removal of the solvent in vacuo afforded the crude desired title compound as an orange solid.

Reference Example 3F. 10-Amino-4-methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.7)

Methyl 10-fluoro-4-methoxy-benzo[a]phenazine-11-carboxylate (IV.5) (466 mg) and sodium azide (900 mg) were dissolved in N, N-di The mixture in methylformamide (10 mL) was heated for 18 hours. The reaction mixture was then cooled, diluted with water, and sodium hydroxide solution was added to give a brown precipitate. The precipitate was collected by filtration and washed with water and ether to give the desired title compound as a yellow solid (242mg).

NMR, CDCl3, 4.08(3H, s), 4.20(3H, s), 7.25(1H, d), 7.70-7.72(2H, m), 7.90(1H, d), 8.36(1H, d), 8.52( 1H,d), 8.85(1H,d).

MS DCI/NH3 m/z 334 (MH+, 100%)

Embodiment 1: the preparation of the compound of general formula (I)

Example 1A. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

At room temperature, a mixture of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) (129 mg) and 1,1'-carbonyldiimidazole (138 mg) was dissolved in anhydrous N, N - Stir in dimethylformamide (8 mL) for 4 hours. To this mixture was added N,N-dimethylethylenediamine (commercial) (0.5 mL) and the resulting reaction mixture was further stirred at room temperature for 30 minutes. Volatiles were removed in vacuo. The residue was dissolved in dichloromethane, washed with water, dried ( _MgSO4 ), and the solvent was removed in vacuo to give the crude product. The resulting product was purified using flash chromatography (5% methanol in dichloromethane) to give the title compound as a bright yellow solid (120mg).

In a similar manner, starting from the appropriate acid of formula (II) and amine of formula (III), various compounds of the following formula (I) were prepared:

From a mixture of 4-nitro-benzo[a]phenazine-11-carboxylic acid (II.27) and 3-nitro-benzo[a]phenazine-11-carboxylic acid (II.28) with N,N - Preparation of 4-nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide from dimethylethylenediamine followed by purification using flash chromatography;

Preparation of 4-benzyloxy-benzo[a]phenazine-11 from 4-benzyloxy-benzo[a]phenazine-11-carboxylic acid (II.13) and N,N-dimethylethylenediamine - formic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-prop-2-ynyloxy from 4-prop-2-ynyloxy-benzo[a]phenazine-11-carboxylic acid (II.14) and N,N-dimethylethylenediamine - Benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-ethoxy-benzo[a]phenazine-11 from 4-ethoxy-benzo[a]phenazine-11-carboxylic acid (II.11) and N,N-dimethylethylenediamine - formic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-isobutoxy-benzo[a]phenazine from 4-isobutoxy-benzo[a]phenazine-11-carboxylic acid (II.15) and N,N-dimethylethylenediamine -11-Formic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-(4-chloro-benzyloxy)-benzo[a]phenazine-11-carboxylic acid (II.16) and N,N-dimethylethylenediamine base)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-(2-methoxy ((2-dimethylamino-ethyl)-amide;

Preparation of [11-(2-dimethylamino-ethyl Carbamoyl)-benzo[a]phenazin-4-yloxy]ethyl acetate;

From 4-[2-(tert-butyl-dimethyl-silyloxy)-ethoxy]-benzo[a]phenazine-11-carboxylic acid (II.19) and N,N-dimethylethyl Preparation of 4-[2-(tert-butyl-dimethyl-silyloxy)-ethoxy]-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide from diamines ;

Preparation of 4-methyl-benzo[a]phenazine-11-carboxylic acid from 4-methyl-benzo[a]phenazine-11-carboxylic acid (II.2) and N,N-dimethylethylenediamine (2-Dimethylamino-ethyl)-amide;

Preparation of 4-fluoro-benzo[a]phenazine-11-carboxylic acid (2 - dimethylamino-ethyl)-amide;

Preparation of 4-methylthio-benzo[a]phenazine-11 from 4-methylthio-benzo[a]phenazine-11-carboxylic acid (II.12) and N,N-dimethylethylenediamine - formic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-bromo-benzo[a]phenazine-11-carboxylic acid (II.5) and N,N-dimethylethylenediamine from 4-bromo-benzo[a]phenazine-11-carboxylic acid (2 - dimethylamino-ethyl)-amide;

Preparation of 4-cyano-benzo[a]phenazine-11-carboxylic acid from 4-cyano-benzo[a]phenazine-11-carboxylic acid (II.6) and N,N-dimethylethylenediamine (2-Dimethylamino-ethyl)-amide;

Preparation of 4-methylsulfonyl-benzo[a]phenazine-11 from 4-methylsulfonyl-benzo[a]phenazine-11-carboxylic acid (II.8) and N,N-dimethylethylenediamine - formic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-chloro-benzo[a]phenazine-11-carboxylic acid (II.7) and N,N-dimethylethylenediamine from 4-chloro-benzo[a]phenazine-11-carboxylic acid (2 - dimethylamino-ethyl)-amide;

Preparation of 4-azido-benzo[a]phenazine-11 from 4-azido-benzo[a]phenazine-11-carboxylic acid (II.9) and N,N-dimethylethylenediamine -Formic acid (2-dimethylamino-ethyl)-amide; formed from benzo[a]phenazine-4,11-dicarboxylic acid 4-methyl ester (II.10) and N,N-dimethylethylenediamine Preparation of methyl 11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylate;

From 4-methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.21) and 3-methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.22) Preparation of 4-methylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide trifluoroacetate from mixture with N,N-dimethylethylenediamine , the two isomers separated by preparative HPLC;

From 4-dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II.23) and 3-dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (II. 24) mixture with N,N-dimethylethylenediamine to prepare 4-dimethylsulfamoyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using The two isomers separated by flash chromatography;

Preparation of 9-bromo-4-methoxy-benzene from 9-bromo-4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.35) and N,N-dimethylethylenediamine And[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

From a mixture of 4-nitro-benzo[a]phenazine-11-carboxylic acid (II.27) and 3-nitro-benzo[a]phenazine-11-carboxylic acid (II.28) with 1-di Preparation of 4-nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide from methylamino-2-propylamine and purification of the resulting product using flash chromatography;

Example 1B. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide

A mixture of 4-methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.1) (350 mg) and 1-dimethylamino-2-propylamine (2 mL) (commercial) was dissolved under _N Heat to 110°C for 4 hours. The reaction mixture was then cooled and excess amine was removed in vacuo. The residue was purified using flash chromatography (silica gel, ethyl acetate, then 25% methanol in ethyl acetate) to afford the title compound as a yellow solid (164 mg).

Following a similar procedure, starting from the appropriate esters of formula (IV) and amines of formula (III), the following compounds of formula (I) were prepared:

Preparation of 4-methoxy-benzo[a]phenazine from methyl 4-methoxy-benzo[a]phenazine-11-carboxylate (IV.1) and 3-(dimethylamino)propylamine (commercial) Oxyzine-11-carboxylic acid (3-dimethylamino-propyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylate methyl ester (IV.1) and 1,3-diamino-2-hydroxypropane (commercial) a] phenazine-11-carboxylic acid (3-amino-2-hydroxy-propyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.1) and N ² ,N ² -dimethyl-propane-1,2-diamine (commercial product) Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-propyl)-amide;

From 4-dimethylamino-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.3) and 3-dimethylamino-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.4) Preparation of 4-dimethylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide from a mixture with N,N-dimethylethylenediamine, followed by purification using flash chromatography Remove a small amount of isomers;

Preparation of 4-methoxy-10- from 4-methoxy-10-methylamino-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.6) and N,N-dimethylethylenediamine Methylamino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 10-amino-4-methoxy from 10-amino-4-methoxy-benzo[a]phenazine-11-carboxylic acid methyl ester (IV.7) and N,N-dimethylethylenediamine - Benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-propyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylate (IV.1) and N-methylethylenediamine from 4-methoxy-benzo[a]phenazine-11- Formic acid (2-methylamino-ethyl)-amide;

Example 1C. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1,1-dimethyl-ethyl)-amide

A mixture of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and thionyl chloride was heated to reflux for 6 minutes. Thionyl chloride was then removed in vacuo. The residue was dissolved in anhydrous dichloromethane at 0°C, and 1-dimethylamino-2-methyl-2-aminopropane (commercial) was added. After stirring for 2 hours, the reaction mixture was dissolved in dichloromethane, washed with sodium bicarbonate solution, dried ( _MgSO4 ), and the solvent was removed in vacuo to give the crude product. The resulting product was purified using flash chromatography to afford the title compound.

In a similar manner, starting from the appropriate acid of formula (II) and amine of formula (III), various compounds of the following formula (I) were prepared:

Preparation of 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (II.36) and N,N-dimethylethylenediamine from 4,10-dimethoxy-benzo[a] a] phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4-methoxy-8-methyl from 4-methoxy-8-methyl-benzo[a]phenazine-11-carboxylic acid (II.37) and N,N-dimethylethylenediamine - Benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 9-chloro-4-methoxy-benzene from 9-chloro-4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.38) and N,N-dimethylethylenediamine And[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Preparation of 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (II.36) and 1-dimethylamino-2-propylamine (commercial product) from 4,10-dimethoxy-benzene And[a]phenazine-11-carboxylic acid (2-dimethylamino-1-methyl-ethyl)-amide;

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and N ¹ , N ¹ -dimethyl-butane-1,2-diamine Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-propyl)-amide from hydrochloride (III.2);

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 3N ¹ , N ¹ -trimethyl-butane-1,2-diamine Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-methyl-propyl)-amide from hydrochloride (III.3);

From 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and N ¹ ,N ¹ -dimethyl-3-phenyl-propane-1, in the presence of triethylamine, Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-phenyl-ethyl)-amide from 2-diamine hydrochloride (III.4);

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and (S)-N ¹ , N ¹ -dimethyl-propane-1,2 - Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-(S)-methyl-ethyl)-amide from diamine hydrochloride (III.5) ;

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and (R)-N ¹ , N ¹ -dimethyl-propane-1,2 - preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(R)-methyl-ethyl)-amide from diamine hydrochloride (III.6);

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and (S)-2-amino-3-dimethylamino-propan-1-ol Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(S)-hydroxymethyl-ethyl)-amide from hydrochloride (III.7);

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 3(S)-amino-4-dimethylamino-butan-2(S) Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (1(S)-dimethylaminomethyl-2(S)-hydroxy-propyl) from -alcohol hydrochloride (III.8) - amides;

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 3-amino-4-dimethylamino-butan-1-ol hydrochloride ( III.9) Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid [1-dimethylamino-1-(2-hydroxyethyl)]-acetamide;

Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 1-(2-aminoethyl)piperidine (commercial) from 4-methoxy-benzo[a] Phenazine-11-carboxylic acid (2-piperidin-1-yl-ethyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 4-(2-aminoethyl)morpholine (commercial) from 4-methoxy-benzo[a] Phenazine-11-carboxylic acid (2-morpholin-4-yl-ethyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 1-(aminoethyl)pyrrolidine (commercial) -11-Formic acid (2-pyrrolidin-1-yl-ethyl)-amide;

Preparation of 4-methoxy-benzo[a]phenazine from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and N,N-diethylethylenediamine (commercial) Oxyzine-11-carboxylic acid (2-diethylamino-ethyl)-amide;

Preparation of 4-methoxy-benzene from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and N,N-bis(2-hydroxyethyl)ethylenediamine (commercial) And[a]phenazine-11-carboxylic acid {2-[bis(2-hydroxy-ethyl)-amino]-ethyl}-amide;

In the presence of aqueous sodium carbonate solution, from 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (II.36) and (S)-2-amino-3-dimethylamino-propane- Preparation of 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(S)-hydroxymethyl-ethyl )-amide;

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and 1-methyl-3-(R)-aminopyrrolidine hydrochloride (III .10) Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (1-methyl-pyrrolidin-3-(R)-yl)-amide;

In the presence of triethylamine, from 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and (R)-3N ¹ , N ¹ -trimethyl-butane-1, Preparation of (R)-4-methoxy-benzo[a]phenazine-11-carboxylic acid (1-dimethylaminomethyl-2-methyl- Propyl)-amide;

In the presence of triethylamine, trihydrochloride (III.11);

From 4-methoxy-benzo[a]phenazine-11-carboxylic acid (II.1) and N ¹ , N ¹ , N ² , N ² -tetramethyl-propane-1,2,3-triamine Preparation of 4-methoxy-benzo[a]phenazine-11-carboxylic acid (2,3-bis)-dimethylamino-propyl)amide;

Example 2: Interconversion of compounds of formula (I)

Compounds of formula (I) prepared as described in Example 1 were converted into other compounds of formula (I) as described below.

Example 2i. 4-Hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide: hydrobromide

To 4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (I.1) (727mg) in anhydrous dichloromethane cooled to -5°C (15 mL) was added a 1.0 M solution of boron tribromide in dichloromethane (13.6 mL). After stirring for 4 hours, the reaction mixture was poured into an ice/water mixture to give a precipitate, which was collected by filtration. The precipitate was triturated in a hot methanol/ethyl acetate mixture to give the title compound as a light brown solid (505 mg).

Example 2iii. 4-Cyanomethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To 4-hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide hydrobromide (230mg) in anhydrous N,N-dimethylformamide (3mL) Potassium tert-butoxide (175 mg) was added to the suspension in , followed by bromoacetonitrile (47 μL). The resulting reaction mixture was heated to 100°C for 1 hour. The reaction mixture was then cooled, diluted with ethyl acetate, washed with sodium carbonate solution and brine, dried ( _MgSO4 ) and the solvent removed in vacuo to give the crude product. The resulting product was purified using flash chromatography (silica gel, 25% MeOH in ethyl acetate) to afford the title compound as a yellow solid (74 mg).

In a similar manner, 4-hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide hydrobromide and an appropriate alkylating agent were used to prepare Various compounds:

Preparation of 4-(pyrimidin-2-yloxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using 2-bromopyridine;

Preparation of 4-(2-morpholin-4-yl-ethoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino -ethyl)-amide;

Preparation of 4-(3-cyano-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using 4-bromobutyronitrile;

Preparation of 4-(3-Dimethylamino-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using 3-dimethylaminopropyl chloride hydrochloride ;

Preparation of 4-carbamoylmethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using 2-bromoacetamide;

Preparation of 4-(2-oxo-propoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using chloroacetone; and

[11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yloxy]acetic acid tert-butyl ester was prepared using tert-butyl bromoacetate.

Example 2iv. 11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yl ethyl carbamate

4-Hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide hydrobromide (540 mg), triethylamine (0.51 mL) and ethyl isocyanate ( 0.29 mL) was stirred in anhydrous N,N-dimethylformamide (3 mL). The product slowly precipitated from the reaction mixture and was collected by filtration and washed with diethyl ether to give the title compound as a yellow solid (210 mg).

Example 2v. 11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yl acetate

4-Hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide hydrobromide (45 mg), triethylamine (71 μL) and acetyl chloride ( 20 μL) in dichloromethane (1.4 mL) was stirred for 2 hours. All volatiles were removed in vacuo and the resulting residue was purified using column chromatography (10% methanol in dichloromethane) to afford the title compound as a yellow solid (27mg).

Example 2vi. [11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yloxy]acetic acid trifluoroacetate

To [11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazin-4-yloxy]acetate tert-butyl ester (18 mg) in anhydrous dichloromethane (1 mL) Trifluoroacetic acid (1 mL) was added to the solution. After stirring for 4 hours, the solvent was removed in vacuo to give the crude product. Trituration with diethyl ether afforded the title compound as a yellow solid (10 mg).

Example 2vii. Benzo[a]phenazine-4,11-dicarboxylic acid 4-amide 11-[(2-dimethylamine-ethyl)-amide]trifluoroacetate

Methyl 11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylate (200 mg) was sonicated in methanol (20 mL) to give a fine suspension. Sodium cyanide (22 mg) was added thereto. The mixture was then sprayed with ammonia for 15 minutes. The reaction mixture was stirred at room temperature for 10 days, and the mixture was sprayed with ammonia every day. After 10 days, the volatiles were removed in vacuo and the resulting residue was purified using flash chromatography to give the crude product. The crude product was further purified using preparative HPLC (isotropic; 80:20 _H2O /acetonitrile) to afford the title compound as a yellow solid (10 mg).

Example 2viii.11-(2-Dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylic acid trifluoroacetate

Methyl 11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylate (200 mg) was sonicated in a mixture of methanol (4 mL) and ammonium hydroxide (20 mL) . The suspension was heated to 50°C for 92 hours. Subsequent removal of all volatiles in vacuo afforded a crude product which was purified using preparative HPLC to afford the title compound (20 mg).

Example 2ix. 4-Hydroxymethyl-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

At 0°C, 11-(2-dimethylamino-ethylcarbamoyl)-benzo[a]phenazine-4-carboxylic acid methyl ester (317mg) in tetrahydrofuran (18mL) and 2-propanol (10mL ) was added lithium borohydride (2.0 M solution in tetrahydrofuran, 1.97 mL). The reaction mixture was stirred overnight at room temperature, then quenched with ammonium chloride solution. The reaction mixture was extracted with ethyl acetate, dried ( _MgSO4 ) and concentrated in vacuo. The resulting residue was purified using flash chromatography (10% MeOH in dichloromethane) to afford the title compound as a yellow solid (98 mg).

Example 2x. 4-(N-Hydroxycarbamimidoyl)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

4-Cyano-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (20 mg), potassium carbonate (37 mg) and hydroxylamine hydrochloride (19 mg) were dissolved in ethanol (5 mL) The mixture was heated to reflux for 18 hours. The reaction mixture was filtered, the filtrate was collected and the solvent was removed in vacuo to afford the title compound (20 mg).

Example 2xi. 4-Dimethylaminomethyl-3-hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

3-Hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (26 mg) was sonicated in acetic acid (2 mL) to give a fine suspension. Thereto were added 40% aqueous dimethylamine (3 mL) and 37% aqueous formaldehyde (3 mL). The reaction mixture was stirred for 2 days. The volatiles were removed in vacuo to give the title compound (29 mg).

Following a similar procedure, 3-dimethylaminomethyl-4-hydroxy-benzo[a] was prepared from 4-hydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide ] phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide.

Example 2xii. 4-(2-Hydroxy-ethoxy)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To 4-[2-(tert-butyl-dimethyl-silyloxy)-ethoxy]-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (125mg ) in tetrahydrofuran (5 mL) was added 1.0 M tetrabutylammonium fluoride solution (1.2 mL). After stirring for 1.5 hours the reaction mixture was diluted with ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a crude product which was purified using flash chromatography to give the title compound as an orange solid (24 mg).

Example 2xiii. 4-Amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

4-Nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (176mg), indium (154mg) and saturated ammonium chloride solution (5mL) in ethanol (20mL ) was heated to reflux for 3 hours. The reaction mixture was cooled, quenched with water and filtered through celite. The resulting filtrate was concentrated in vacuo and the resulting residue was treated with sodium bicarbonate solution, extracted into chloroform, dried ( _MgSO4 ) and the solvent removed in vacuo to afford the title compound as a red solid (163 mg). Following a similar procedure, 3-amino-benzo[a]phenazine-11-carboxylic acid was prepared from 3-nitro-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (2-Dimethylamino-ethyl)-amide.

Example 2xiv. 4-Acetamido-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To a solution of 4-amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (20 mg) in tetrahydrofuran (5 mL) was added pyridine (0.1 mL) and acetyl chloride ( 20 μL). After stirring for 1 h, the reaction mixture was extracted into ethyl acetate, washed with aqueous sodium bicarbonate, dried ( _MgSO4 ) and the solvent was removed in vacuo. The residue was triturated with ether to give the title compound as a yellow solid (10 mg).

Following a similar procedure, 3-acetamido-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide and acetyl chloride were prepared from 3-amino-benzo[a]phenazine- 11-Formic acid (2-dimethylamino-ethyl)-amide;

Following a similar procedure, 4-methylsulfonylamino-benzo[a] was prepared from 4-amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide and methanesulfonyl chloride Phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Following a similar procedure, 3-methylsulfonylamino-benzo[a] was prepared from 3-amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide and methanesulfonyl chloride Phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide;

Following a similar procedure, 4 was prepared from 4-amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide using an excess of methanesulfonyl chloride and triethylamine as a base. -Bis-(methylsulfonylamino)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide.

Example 2xv. 4-(cyanomethyl-amino)-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To a solution of 4-amino-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (69 mg) in methanol (10 mL) was added formaldehyde (37% solution, 1.0 mL) , potassium cyanide (102 mg) and 2N hydrochloric acid (1.0 mL). The reaction mixture was heated to 50 °C for 3 hours. The reaction mixture was then cooled, diluted with water and sodium bicarbonate solution, extracted into dichloromethane, dried ( _MgSO4 ) and the solvent was removed in vacuo to give the crude product. The resulting crude product was purified using flash chromatography (10% methanol in dichloromethane) to afford the title compound as a purple solid (13 mg).

Example 2xvii. 4,10-Dihydroxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To a cold solution of 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (96 mg) in dichloromethane (2 mL) was added 1.0 A solution of boron tribromide in dichloromethane (2.14 mL, 9 equiv). The reaction mixture was stirred for 16 hours, then ice was added together with sodium carbonate and sodium chloride. The organic phase was extracted into dichloromethane, dried ( _MgSO4 ) and the solvent removed in vacuo to give an orange compound which was recrystallized from dichloromethane/methanol/hexanes to give the title compound (6 mg).

Example 2xviii. 10-Hydroxy-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

To a cold solution of 4,10-dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (300 mg) in dichloromethane (25 mL) was added 1.0 A solution of boron tribromide in dichloromethane (1.63 mL, 2.2 equiv). The reaction mixture was stirred for 6 hours, then ice was added together with sodium carbonate and sodium chloride. The organic phase was extracted into dichloromethane, dried ( _MgSO4 ) and the solvent was removed in vacuo to give a yellow solid which was purified using flash chromatography to give the title compound (61 mg).

Following a similar procedure, 10-hydroxyl -4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(R)-methyl-ethyl)-amide; and

Following a similar procedure, 10- Hydroxy-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1(S)-hydroxymethyl-ethyl)-amide.

Example 2xix. 4-Methoxy-9-methylthio-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

9-Chloro-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (85mg) and sodium thiomethoxide (43mg) were dissolved in N,N- The mixture in dimethylformamide (1 mL) was heated to 120°C for 6 hours and 60°C for 16 hours. The reaction mixture was then cooled, diluted with ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a yellow solid which was purified using flash chromatography to give the desired title compound (37mg).

Example 2xx. 4,9-Dimethoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide

A solution of 9-chloro-4-methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-ethyl)-amide (85 mg) and 25% sodium methoxide in methanol (4 mL) The mixture was heated to reflux for 6 hours. The reaction mixture was then cooled, diluted with ethyl acetate, washed with water, dried ( _MgSO4 ) and the solvent removed in vacuo to give a yellow solid which was purified using flash chromatography to give the desired title compound (42 mg).

Example 2xxi. 4-Methoxy-benzo[a]phenazine-11-carboxylic acid (2-dimethylamino-1-hydroxymethyl-ethyl)-amide

3-Dimethylamino-2-[(4-methoxy-benzo[a]phenazine-11-carbonyl)-amino]-propionic acid methyl ester (335 mg) and lithium borohydride (72 mg ) in tetrahydrofuran (10 mL) and isopropanol (10 mL) was stirred for 18 hours. Another 5 equivalents of lithium borohydride were added and the resulting mixture was stirred for a further 18 hours. The reaction was quenched with ammonium chloride solution and extracted with ethyl acetate, dried ( _MgSO4 ) and the solvent removed in vacuo to give a brown gum. The resulting product was purified using flash chromatography and triturated with diethyl ether to give the desired title compound as an orange powder.

Example 3: Biological testing of compounds of formula (I)

Cytotoxicity of compounds of formula (I) was determined using the H69 parental (H69/P) human small cell lung cancer cell line and the drug-resistant human small cell lung cancer cell line H69/LX4 overexpressing P-glycoprotein (Pgp). Cytotoxicity in the H69/LX4 cell line determined by the IC ₅₀ (concentration required to kill 50% of the cells) divided by the cytotoxicity in the H69/P cell line indicates that the compound is affected by the Pgp-dependent MDR The degree is called the resistance factor (Rf) of the compound.

H69/P and H69/LX4 cells were transferred to 96-well tissue culture plates by pipette and incubated at 37°C for 4 hours. Compounds of formula (I) or standard TAS-103, doxorubicin and topotecan were then added at concentrations ranging from 0.01 nM to 5 μM. The plates were incubated for 5-6 days before AlamarBlue was added to the wells, and the plates were returned to the incubator for 5-8 hours to develop color. The number of cells in the culture plate at the end of this period is directly proportional to the absorbance measured at a wavelength of 570 nm (reference wavelength 600 nm).

Compounds of formula (I) are active in the range of 5 nM to 5 μM. Specific results are listed in Table 1 for selected compounds.

Table 1 compound H69/P IC ₅₀ (nM) H69/LX4 IC ₅₀ (nM) Rf TAS-103 twenty one twenty two 1.1 Doxorubicin 27.3 3700 135 Topotecan 15.9 61.5 3.9 4 35 48 1.4 3 35 49 1.4 11 28 twenty four 0.9 94 19 25 1.3 68 19 25 1.3 75 twenty three 28 1.2 81 20 25 1.2 78 twenty four 31 1.3 98 twenty one 20 1 84 19 19 1

The cytotoxicity of the compounds described here was also tested in the COR-L23 parental (COR-L23/P) human NSCLC cell line and drug-resistant human NSCLC overexpressing multidrug resistance-associated protein (MRP). The lung cancer cell line COR-L23/R was used for measurement. Cytotoxicity in the L23/R cell line determined by the IC ₅₀ (concentration required to kill 50% of the cells) divided by the cytotoxicity in the L23/P cell line indicates that the compound is affected by the MRP-dependent MDR The degree is called the resistance factor (Rf) of the compound.

L23/P and L23/R cells were transferred to 96-well tissue culture plates by pipette and incubated at 37°C for 4 hours. Compounds of formula (I) or standard TAS-103, doxorubicin and topotecan were then added at concentrations ranging from 0.01 nM to 5 μM. The plates were incubated for 5-6 days before cell proliferation was assessed using the sulphurhodamineB (SRB) assay (Skehan et al; J Natl Cancer Inst 1990, 82, pp. 1107-1112).

Compounds are active in the range of 1 nM to 5 μM. Specific examples are listed in Table 2.

Table 2 compound L23/P IC ₅₀ (nM) L23/R IC ₅₀ (nM) Rf TAS-103 16.3 twenty two 1.3 Doxorubicin 20.3 326.8 16.1 Topotecan 13.6 20.8 1.5 4 14.7 16.8 1.1 3 14.4 19.9 1.4 11 6.1 17.7 2.9 94 5.7 3.8 0.7 68 13.1 44.4 3.4 75 13.0 17.2 1.3 81 4 12.4 3.1 78 7.6 8.9 1.2 98 9.6 8.8 0.9

The cytotoxicity of the compounds described here was also tested with the Jurkat human leukemia cell line (JL _c ), the amsacridine-resistant Jurkat human leukemia cell line (JL _A ) and the doxorubicin-resistant Jurkat human leukemia cell line Department ( _JLD ) to determine. Cytotoxicity in the JL _A or JL _D cell lines divided by the cytotoxicity in the JL _c cell line determined by the _IC50 (concentration required to kill 50% of the cells) indicates that the compound is affected by atypical drug resistance The degree is called the resistance factor (Rf) of the compound. The method used has been described previously (Finlay et al., Eur J. Cancer 32A, 708-714, 1996). Compounds are active in the range of 1 nM to 5 μM. Specific examples are listed in Table 3.

table 3 compound JL _C IC ₅₀ (nM) JL _A IC ₅₀ (nM) Rf(JL _A /JL _C ) JL _D IC ₅₀ (nM) Rf(JL _D /JL _C ) TAS-103 5.4 302 55.9 384 71.1 Doxorubicin 7.0 25.9 3.7 109 15.6 4 19.0 26.6 1.4 22.8 1.2 3 27.0 21.6 0.8 24.3 0.9 2 37 96 2.6 107 2.9 35 28 19 0.7 25 0.9 78 twenty one 14 0.7 17 0.8 81 8.7 9.2 1.1 9.3 1.1 84 4.4 9.8 2.2 7.2 1.6 87 16 16 1.0 17 1.0 94 9.2 16 1.8 14 1.6 98 8.6 18 2.1 14 1.6

The ability of compounds to stabilize cleavable complexes in the presence of topoisomerase I or II was also investigated essentially as previously described (Finlay et al., Eur J. Cancer 32A, 708-714, 1996). The presence of a cleavable complex is indicated by an increase in the number and intensity of bands observed after electrophoresis and autoradiography. Results are expressed in terms of the effective concentration range over which an increase in cleavable complex was observed relative to the control in the absence of drug. Various compounds described here were tested using these methods and compounds that showed toxicity to topoisomerase I and II in the range of 0.01-20 [mu]M. Specific examples are listed in Table 4.

Table 4 Effective concentration range (μM) compound topoisomerase I topoisomerase II TAS-103 0.3-10.0 0.3-10.0 3 0.1-3.0 0.1-3.0 4 0.1-3.0 0.1-3.0 35 0.03-1.0 0.03-1.0 78 0.03-1.0 0.03-1.0 84 0.03-1.0 0.03-1.0

Embodiment 4: pharmaceutical composition

Tablets weighing 0.15 g and containing 25 mg of a compound of the invention can be prepared as follows:

Composition for 10,000 tablets

Compound of the present invention (250g)

Lactose (800g)

Cornstarch (415g)

Talc powder (30g)

Magnesium Stearate (5g)

Mix together the compound of the invention, lactose and half of the cornstarch. The mixture is then forced through a 0.5 mm mesh size sieve. Corn starch (10 g) was suspended in warm water (90 mL). The powder is granulated with the resulting paste. The resulting granules were dried and broken into small pieces on a 1.4 mesh size sieve. The remaining amount of starch, talc and magnesium stearate are added, mixed carefully and processed into tablets.

Example 5: Characterization of compounds of formula (I)

The compound prepared in Example 3 was characterized by proton NMR spectrum and mass spectrum. All proton NMRs were performed at 400 MHz. Perform mass spectrometric characterization using desorption chemical ionization or electrospray ionization. The results are listed in the table below. Compound number molecular formula mass spectrum data ¹ H NMR data 3 C22H22N4O2 MH+@375 DMSO; 10.27(br, 1H), 8.96(d, J=8.19Hz, 1H), 8.71(dd, J=7.20, 1.48Hz, 1H), 8.47(d, J=9.5Hz, 1H), 8.41(dd , J=8.19, 1.46Hz, 1H), 8.10(dd, J=7.14, 1.24Hz, 1H), 7.93(d, J=9.63Hz, 1H), 7.85(t, J=8.14Hz, 1H), 7.46 (d, J=7.99Hz, 1H), 4.08(s, 3H), 3.71(q, J=5.56Hz, 2H), 2.63(t, J=5.95Hz, 2H), 2.34(s, 6H) 4 C21H20N4O2, HBr M+@360 DMSO; 2.91(6H,s), 3.45(2H,t), 3.97-4.02(2H,m), 7.37(1H,d), 7.72-7.78(1H,m), 7.92(1H,d), 8.08- 8.12(1H, m), 8.45-8.51(2H, m), 8.54-8.61(2H, m), 9.42(1H, br.), 10.13(1H, t), 10.67(1H, s). 7 C21H19N5O3 DCI+NH3, MH+, 390 (CDCL3), 2.50 (6H, s, 2xCH3), 2.75 (2H, t, CH2), 3.95 (2H, q, CH2), 7.95 (1H, t, ArH), 8.10 (1H, t, ArH), 8.30 (1H, d, ArH), 8.45 (1H, d, ArH), 8.50 (1H, d, ArH), 8.70 (1H, d, ArH), 9.10 (1H, dd, ArH), 9.90 (1H, d, ArH), 10.50 (1H, Br., NH). 10 C22H21BrN4O2 MH+453/455 (1:1) (Chloroform): 10.73 (br, t, 1H), 8.92 (dd, 1H), 8.55 (t, 1H), 8.38 (dd, 1H), 8.04-7.90 (m, 3H), 7.10 (d, 1H), 4.09(s, 3H), 3.85(q, 2H), 2.74(t, 2H), 2.33(s, 6H) 11 C23H21N5O2 MSDCI/NH3m/e 400 (MH+, 100%) CDCl3; 2.36 (6H, s), 2.68 (2H, t, J = 5.9Hz), 3.81-3.84 (2H, m), 5.02 (2H, s), 7.30 (1H, d = 8.1Hz), 7.71-7.75 (1H, m), 7.92-7.98 (2H, m), 8.36-8.41 (2H, m), 8.96 (1H, dd, J=7.2, 1.5Hz), 9.15 (1H, d, J=8.3Hz), 10.70 (1H, broad peak). 12 C28H26N4O2 MSDCI/NH3m/e 451 (MH+, 100%) CDCl3; 2.42 (6H, s), 2.75 (2H, t, J=6.0Hz), 3.89-3.93 (2H, m), 5.35 (2H, s), 7.33-7.49 (4H, m), 7.55 (2H, d, J = 7.2Hz), 7.73-7.77 (1H, m), 7.95-8.0 (2H, m), 8.40 (1H, dd, J = 8.6, 1.5Hz), 8.63 (1H, d, J = 9.4Hz ), 9.00-9.07 (2H, m), 10.90 (1H, broad peak). 13 C24H22N4O2 MSDCI/NH3m/e 399 (MH+, 100%) CDCl3; 2.37(6H, s), 2.55(1H, t, J=2.4Hz), 2.70(2H, t, J=6.0Hz), 3.82-3.87(2H, m), 4.95(2H, d, J= 2.4Hz), 7.32(1H, d, J=8.3Hz), 7.70-7.74(1H, m), 7.90-7.95(2H, m), 8.35(1H, dd, J=8.50, 1.6Hz), 8.50( 1H, d, J=9.8Hz), 8.95(1H, dd, J=7.30, 1.5Hz), 9.01(1H, d, J=8.2Hz), 10.80(1H, broad peak). 15 C23H24N4O2 MSDCI/NH3m/e 389 (MH+, 100%) d6-DMSO; 1.52 (3H, t, J = 6.9Hz), 2.35 (6H, s), 2.65 (2H, t, J = 5.9Hz), 3.72-3.75 (2H, m), 4.30 (2H, q, J=6.9Hz), 7.43(1H, d, J=7.9Hz), 7.80-7.84(1H, m), 7.92(1H, d, J=9.5Hz), 8.05-8.10(1H, m), 8.42( 1H,dd,J=8.6,1.6Hz), 8.45(1H,d,J=9.5Hz), 8.72(1H,dd,J=7.1,1,5Hz), 8.94(1H,d,J=8.2Hz) , 10.25(1H,t,J=5.0Hz). 16 C25H28N4O2 EI+ m/z 416 (M+, 12%); 346 (100%); 329 (58%); CDCl3; 10.87 (1H, br); 8.93 (2H, m); 8.54 (1H, d, J = 9.5Hz); 8.35 (1H, dd, J = 8.4, 1.5Hz); 7.89 (2H, m); 7.69 (1H, t, J=8.0Hz); 7.19(1H, m); 3.93(2H, d, J=6.4Hz); 3.87(2H, m); 2.75(2H, m); 2.40(6H, s) ;2.24(1H,m);1.11(6H,d,J=6.8Hz). 17 C28H25ClN4O2 DEI m/e484/486 3:1 (M+, 100) d6-DMSO; 2.38 (6H, s); 2.62-2.70 (2H, m); 3.70-3.77 (2H, m); 5.44 (2H, s); 7.52 (2H, d, J=8.3Hz); 7.58 ( 1H, d, J = 8.0Hz); 7.64 (2H, d, J = 8.3Hz); 7.88 (1H, t, J = 8.1Hz); 8.01 (1H, d, J = 9.5Hz); 8.10 (1H, t, J = 7.8Hz); 8.46 (1H, d, J = 8.4Hz); 8.56 (1H, d, J = 9.5Hz); 8.73 (1H, d, J = 7.0Hz); 9.03 (1H, d, J=8.0Hz); 10.22-10.30 (1H, broad peak). 18 C24H26N4O3 DCI+NH3 CDCl3; 2.69 (6H, s); 2.92-3.10 (2H, m); 3.49 (3H, s); 3.83-3.93 (2H, t, J=4.4Hz); 3.94-4.10 (2H, m); m/e419 (M+, 100) 4.28-4.40 (2H, t, J = 4.4Hz); 7.22 (1H, d, J = 8.0Hz); 7.80 (1H, t); 7.84-7.96 (2H, m, J = 4.7Hz, 8.3Hz); 8.37 (1H, d, J = 8.2Hz); 8.54 (1H, d, J = 9.4Hz); 8.82 (1H, d, J = 7.4Hz); 8.90 (1H, d, J = 6.9Hz); 11.10 ( 1H, broad peak). twenty one C23H24N4O3 CI+(NH3) m/z 405(MH+, 100%) CDCl3; 10.69 (1H, br); 8.93 (1H, dd, J = 7.3, 1.5Hz); 8.80 (1H, d, J = 8.3Hz); 8.47 (1H, d, J = 9.6Hz); 8.32 (1H , dd, J = 8.3, 1.5Hz); 7.91 (1H, m); 7.85 (1H, d, J = 9.6Hz); 7.57 (1H, t, J = 8.1Hz); 7.11 (1H, d, J = 7.9Hz); 4.27(2H, m); 4.11(2H, m); 3.83(2H, q, J=5.7Hz); 2.68(2H, t, J=6.0Hz); 2.35(6H, s). twenty two C25H22N6O2 DCI/NH3m/z 439 (MH+, 100%) CDCl3; 2.40(6H, s), 2.72(2H, t, J=5.90Hz), 3.85-3.89(2H, m), 7.05(1H, t, J=4.80Hz), 7.63(1H, d, J= 7.75Hz), 7.80-7.95(3H, m), 8.18(1H, d, J=9.5Hz), 8.36(1H, dd, J=8.4, 1.4Hz), 8.55(2H, d, J=4.80Hz) , 9.00 (1H, dd, J=7.2, 1.3Hz), 9.40 (1H, d, J=8.1Hz), 10.70 (1H, broad peak). twenty three C27H31N5O3 DCI/NH3m/z 474 (MH+, 100%) CDCl3; 2.45(6H, s), 2.68-2.71(4H, m), 2.75(2H, t, J=6.0Hz), 3.01(2H, t, J=5.6Hz), 3.75-3.79(4H, m) , 3.88-3.92(2H, m), 4.36(2H, t, J=5.6Hz), 7.25-7.27(1H, m), 7.71-7.75(1H, m), 7.95-8.01(2H, m), 8.41 (1H, dd, J=8.5, 1.5Hz), 8.55(1H, d, J=9.5Hz), 9.01-9.05(2H, m), 10.8(1H, broad peak). twenty four C25H25N5O2 DCI/NH3m/z 428 (MH+, 100%) CDCl3; 2.35-2.38 (2H, m), 2.45 (6H, s), 2.73-2.75 (4H, m), 3.89-3.93 (2H, m), 4.37 (2H, J=5.7Hz), 7.28-7.30 ( 1H, m), 7.78-7.80 (1H, m), 7.98-8.01 (2H, m), 8.45 (1H, dd, J=8.4, 1.5Hz), 8.56 (1H, d, J=9.7Hz), 9.05 (1H, dd, J=7.2, 1.5Hz), 9.08(1H, d, J=8.2Hz), 10.8(1H, broad peak). 25 C22H22N4O [M+H]+359 (CDCl3) 10.88 (1H, br, s), 9.34 (1H, d, J 7.9), 9.02 (1H, dd, J 7.3, 1.5), 8.42 (1H, dd, J 8.4, 1.5), 8.31 (1H, d, J 9.4), 8.02 (1H, d, J 9.6), 7.98 (1H, d, J 7.3), 7.73 (1H, t, J 7.6), 7.69 (1H, J, 6.9), 3.91 (2H, q , J 5.7), 2.75 (3H, s), 2.60 (2H, t, J 5.7), 2.44 (6H, s) 26 C21H19FN4O [M+H]+363 (CDCl3) 10.67 (1H, s, br), 9.22 (1H, d, J 8.1), 8.97 (1H, dd, J 7.3, 1.5), 8.36 (1H, dd, J 8.4, 1.5), 8.29 (1H, d, J 9.5), 7.97 (1H, d, J 9.5), 7.93 (1H, dd, J 8.5, 7.2), 7.71 (1H, m), 7.47 (1H, m), 3.83 (2H, q, J 5.5 ), 2.68(2H, t, J 5.8), 2.37(6H, s), 27 C26H31N5O2 DCI/NH3m/z 446 (MH+, 100%) CDCl3; 2.15-2.21 (2H, m), 2.34 (6H, s), 2.44 (6H, s), 2.63 (2H, t, J = 7.2Hz), 2.79 (2H, t, J = 6.0Hz), 3.90 -3.95(2H, m), 4.30(2H, t, J=6.3Hz), 7.25-7.27(1H, m), 7.71-7.74(1H, m), 7.95-8.01(2H, m), 8.40(1H , dd, J=8.40, 1.5Hz), 8.55 (1H, d, J=9.5Hz), 8.99-9.03 (2H, m), 10.89 (1H, broad peak). 28 C22H22N4OS EI+ m/z 390 (M+, 14%); 320 (100%); CDCl3; 10.79 (1H, br); 9.32 (1H, m); 9.02 (1H, dd, J = 7.2, 1.5Hz); 8.63 (1H, d, J = 9.7Hz); 8.42 (1H, dd, J = 8.4, 1.5Hz); 8.05 (1H, d, J = 9.7Hz); 7.99 (1H, m); 7.76 (2H, m); 3.90 (2H, m); 2.75 (2H, m); 2.66 (3H, s); 2.43(6H, s) 29 C23H23N5O3 DCI/NH3m/z418 (MH+, 100%) CDCl3 and two drops of D4-MeOH2.32 (6H, s), 2.68 (2H, t, J = 6.2Hz), 3.78 (2H, t, J = 6.2Hz), 4.68 (2H, s), 7.20 (1H, d, J=7.8Hz), 7.70-7.72(1H, m), 7.90-7.95(2H, m), 8.35(1H, dd, J=1.5, 8.5Hz), 8.53(1H, d, J=9.8Hz ), 8.86 (1H, dd, J=7.2, 1.5Hz), 8.95 (1H, d, J=8.3Hz), 3 exchangeable protons (d4) were not observed due to the presence of methanol. 30 C21H20N4O3 DCI/NH3m/z 377 (MH+, 100%) CDCl3; 2.81 (1H, dd, J = 12.6, 7.5Hz), 2.97 (1H, dd, J = 12.6, 3.7Hz), 3.70-3.75 (1H, m), 3.90-4.00 (2H, m), 4.01 ( 3H, s), 7.15(1H, d, J=8.0Hz), 7.65-7.71(1H, m), 7.81(1H, d, J=9.5Hz), 7.85-7.90(1H, m), 8.31(1H , d, J = 7.5Hz), 8.42 (1H, d, J = 9.6Hz), 8.80 (1H, d, J = 8.2Hz), 8.88 (1H, d, J = 6.1Hz), 11.2 (1H, wide peak), OH and NH2 were not observed in NMR 31 C23H24N4O2 DCI+NH3389(MH+) DMSO; 1.92 (2H, t, J = 7.03Hz), 2.2 (6H, s), 2.48 (2H, t, J = Hz), 3.62 (2H, q, J = 6.74Hz), 4.07 (3H, s) , 7.3 (1H, d, J = 8.03Hz), 7.85 (1H, t, J = 8.06), 7.98 (1H, d, J = 9.48), 8.05 (1H, t, J = 7.25Hz), 8.42 (1H , d, J = 8.64Hz), 8.49 (1H, d, J = 9.6Hz), 8.52 (1H, d, J = 6.9Hz), 8.72 (1H, d, J = 8.06Hz), 9.95 (1H, wide peak). 32 C21H19BrN4O CI+m/z 423:425 (1:1, MH+, 98%) CDCl3; 10.61 (1H, br); 9.41 (1H, d, J = 8.0Hz); 8.97 (1H, dd, J = 7.3, 1.6Hz); 8.47 (1H, d, J = 9.8Hz); 8.36 (1H , dd, J=8.4, 1.5Hz); 8.02(2H, m); 7.94(1H, m); 7.61(1H, t, J=7.9Hz); 3.84(2H, m); , J=5.9Hz); 2.37(6H, s). 33 C23H22N4O3 m/z 403(MH+) d6-DMSO; 2.37 (6H, s), 2.52 (3H, s), 2.65 (2H, t, J=6.0), 3.72 (2H, q, J=6.0), 7.74 (1H, d, J=7.0) , 7.99 (1H, t, J=8.0), 8.08 (1H, d, J=9.6), 8.12 (1H, t, J=7.8), 8.31 (1H, d, J=9.6), 8.47 (1H, d , J=8.0), 8.75 (1H, d, J=7.0), 9.38 (1H, d, J=8.0) 34 C24H24N4O3 MSDCI/NH3m/z 417 (MH+, 100%) CDCl3; 2.42(6H, s), 2.45(3H, s), 2.75(2H, t, J=5.9Hz), 3.88-3.91(2H, m), 4.83(2H, s), 7.12(1H, d, J=7.8Hz), 7.71-7.76(1H, m), 7.95-8.02(2H, m), 8.45(1H, d, J=8.4, 1.4Hz), 8.63(1H, d, J=9.6Hz), 9.03(1H, dd, J=7.2, 1.6Hz), 9.15(1H, d, J=8.1Hz), 10.80(1H, broad peak). 35 C23H24N4O2 DCI/NH3 CDCl3; 1.53 (3H, d, J = 6.5Hz), 2.37 (6H, s), 2.55 (1H, dd, J = 12.2, 6.2Hz), 2.84 (1H, dd, J = 12.2, m/z 389 (MH+, 100%) 7.8Hz), 4.10(3H, s), 4.55-4.65(1H, m), 7.28-7.30(1H, m), 7.75-7.80(1H, m), 7.95-8.02(2H, m), 8.42(1H , dd, J=8.4, 1.5Hz), 8.57 (1H, d, J=9.5Hz), 8.81 (1h, d, J=8.2Hz), 9.02 (1H, dd, J=7.3, 1.6Hz), 10.9 (1H, d, J=7.2Hz). 36 C22H19N5O MH+at 370 DMSO; 9.99(t, 1H), 9.62(d, 1H), 8.74(dd, 1H), 8.41(m, 2H), 8.35(d, 1H), 8.24(d, 1H), 8.10-8.00(m, 2H), 3.72(q, 2H), 2.62(t, 2H), 2.35(s, 6H). 37 C24H25N5O3 DCI+NH3432(MH+) d6-DMSO; 0.9 (3H, t, J = 7.4Hz), 2.4 (6H, s), 2.75 (2H, broad peak), 3.37 (2H, t, J = 6.42Hz), 5.3 (2H, broad peak) , 7.57 (1H, d, J = 7.21Hz), 7.75 (1H, t, J = 7.4Hz), 7.91 (2H, t, J = 7.14Hz), 8.13 (1H, d, J = 9.58Hz), 8.3 (1H, d, J = 8.43Hz), 8.95 (1H, d, J = 6.72Hz), 9.21 (1H, d, J = 7.64Hz), 10.76 (1H, broad peak), not one NH observed 38 C21H19N5O3 DCI+NH3; MH+, 390 (CDCL3), 2.50 (6H, s, 2xCH3), 3.10 (2H, t, CH2), 4.05 (2H, q, CH2), 8.10 (1H, dd, ArH), 8.15 (2H, dd, ArH), 8.20 (1H, d, ArH), 8.45 (1H, d, ArH), 8.60 (1H, dd, ArH), 9.0 (1H, dd, ArH), 9.90 (1H, d, ArH), 10.40 (1H, br. , NH) 39 C22H22N4O3S CI+ (NH3) m/z 423 (MH+, 100%) d6-DMSO; 9.79 (1H, br m); 9.64 (1H, d, J = 8.0Hz); 9.10 (1H, d, J = 9.8Hz); 8.65 (1H, m); 8.59 (2H, m); 8.40(1H, d, J=9.8Hz); 8.24(2H, m); 4.04(2H, br m); 3.57(3H, s); 3.53(2H, br t, J=6.3Hz); 2.99(6H , s). 40 C21H19ClN4O DCI+NH3m/e 379 (MH+, 100) CDCl3; 2.38 (6H, s); 2.62-2.70 (2H, t, J = 8.0Hz); 3.78-3.86 (2H, m, J = 5.4, 3.0Hz); 7.69 (1H, t, J = 8.0Hz) ; 7.72 (1H, d, J = 0.9, 6.8Hz); 7.94 (1H, t, J = 1.3, 7.2Hz); 8.04 (1H, d, J = 9.5Hz); 8.37 (1H, d, J = 1.5 , 6.9Hz); 8.51 (1H, d, J = 9.4Hz); 8.98 (1H, d, J = 1.5, 5.6Hz); 9.99 (1H, d, J = 8.0Hz); 10.60 (1H, broad peak) . 41 C21H19N7O [M+H]+386 (CDCl3); 10.75 (1H, br), 9.30 (1H, d, J 8.2), 9.04 (1H, d, J 7.3), 8.42 (2H, t, J 8.4), 8.00 (2H, t, J 7.8) , 7.85 (1H, t, J 9.1), 7.63 (1H, d, J 7.8), 3.90 (2H, q, br), 2.76 (2H, t, br), 2.45 (6H, s) 42 C21H21N5O DCI+NH3, MH+, 360 (CDCL3), 2.50 (6H, s, 2xCH3), 2.90 (2H, t, CH2), 3.95 (2H, m, CH2), 7.10 (1H, d, ArH), 7.65 (1H, t, ArH), 7.90 ( 2H, m, ArH), 8.05 (1H, d, ArH), 8.35 (1H, d, ArH), 8.70 (1H, d, ArH), 8.95 (1H, d, ArH), 11.0 (1H, br., NH) 43 C25H23F3N4O6 DCI/NH3m/z 419 (MH+, 100%) d6-DMSO; 2.70 (6H, s), 3.18 (2H, t), 3.89-3.95 (2H, m), 5.00 (2H, s), 7.42 (1H, d, J=8.0Hz), 7.81-7.86 ( 1H, m), 8.00(1H, d, J=8.0Hz), 8.08-8.12(1H, m), 8.46(1H, d, J=7.8Hz), 8.55-8.62(2H, m), 8.85(1H , d, J=8.1Hz), 10.1 (1H, broad peak). No two acidic OH were observed 44 C23H23N5O2 MH+, 402 (CDCl3), 2.35 (6H, s, 2xCH3), 2.40 (3h, s, CH3), 2.65 (2H, t, CH2), 3.80 (2H, q, CH2), 7.45 (1H, t, ArH), 7.70 (2H, m, ArH), 7.90 (2H, m, ArH), 8.20 (1H, s, NH), 8.25 (1H, d, ArH), 8.70 (1H, d, ArH), 8.80 (1H, d, ArH), 10.10 (1H, br., NH) 45 C23H22N4O3 m/z 403(MH+) (CDCl3); 2.38(6H, s), 2.70(2H, t, J=5.8), 3.87(2H, q, J=5.8), 4.03(3H, s), 7.81(1H, t, J=7.9) , 7.96 (1H, t, J=7.8), 8.07 (1H, d, J=9.8), 8.35-8.40 (2H, m), 9.00 (1H, d, J=7.2), 9.13 (1H, d, J=9.8), 9.68 (1H, d, J=8.1), 10.65 (1H, br) 46 C23H25N5O5S2 DCI+NH3, MH+516 (CDCL3), 2.45 (6H, s, 2xCH3), 2.75 (2H, t, CH2), 3.60 (6H, s, 2xCH3), 3.90 (2H, q, CH2), 7.85 (1H, d, ArH), 7.95 (1H, t, ArH), 8.05 (1H, t, ArH), 8.20 (1H, d, ArH), 8.35 (1H, d, ArH), 8.45 (1H, d, ArH), 9.08 (1H, d, ArH), 9.75 (1H, d, ArH), 10.65 (1H, br, NH). 47 C21H21N5O DCI+NH3; MH+, 360 (CDCl3), 2.45 (6H, s, 2xCH3), 2.90 (2H, t, CH2), 3.95 (2H, q, CH2), 4.45 (2H, br, NH2), 7.10 (1H, dd, ArH), 7.65 (1H, d, ArH), 7.70 (1H, d, ArH), 7.90 (2H, m, ArH), 8.30 (1H, d, ArH), 8.35 (1H, d, ArH), 8.85 (1H, d, ArH), 11.05 (1H, br, NH) 48 C22H22N6O2 MH+403 d6-DMSO; 2.33 (6H, s), 2.63 (2H, t, J=5.91Hz), 3.69-3.78 (2H, m), 6.16 (1H, s), 7.8 (1H, broad singlet), 7.93- 8.12(4H, m), 8.22(1H, broad singlet), 8.42-8.49(1H, m), 8.58(1H, d, J=11.82Hz), 8.72(1H, d, J=7.22Hz), 9.48 -9.52(1H, m), 10.16-10.25(1H, broad multi-peak) 49 C22H22N4O2 CI+ m/z 375 (MH+, 100%) CDCl3; 10.64 (1H, br); 8.92 (1H, dd, J = 7.1, 1.5Hz); 8.80 (1H, dd, J = 7.1, 1.5Hz); 8.24 (2H, m); 7.86 (1H, m) ;7.69(3H,m);5.09(2H,s);3.83(2H,m);2.85(2H,br m);2.50(6H,br s) 50 C24H21F3N4O5 m/z 389(MH+) (d6-DMSO); 2.94 (6H, d, J = 3.7), 3.49 (2H, m), 4.00 (2H, q, J = 6.2), 8.05 (1H, t, J = 7.9), 8.12-8.22 ( 2H, m), 8.48-8.58 (3H, m), 9.17 (1H, d, J=9.8), 9.41 (1H, d, J=8.2), 9.82 (1H, t) 51 C24H24F3N5O5S DCI/NH3m/z438(MH+), 100% CDCl3/d4-MeOD2.67(3H, s), 3.05(6H, s), 3.62(2H, t), 4.25(2H, t), 8.00-8.10(3H, m), 8.45-8.50(2H, m ), 8.88(1H,d), 8.99(1H,d), 9.35(1H,d), 2 NH were not observed due to the presence of MeOD 52 C24H24F3N5O5S DCI/NH3m/e 438(MH+), 100% CDCL3 and MeOD2.62 (3H, s), 3.01 (6H, s), 3.66 (2H, t), 4.13 (2H, t), 7.92-7.99 (1H, m), 8.02-8.08 (3H, m), 8.15(1H,dd), 8.39(1H,dd), 8.80(1H,dd), 9.49(1H,d), no NH observed due to the presence of MeOD, 53 C23H23N5O2 DCI+NH3, MH+402 (CDCl3), 2.32 (3H, s, CH3), 2.40 (6H, s, 2xCH3), 2.80 (2H, t, CH2), 4.05 (2H, q, CH2), 7.75-8.0 (5H, m, ArH) , 8.10 (1H, br., NH), 8.35 (1H, dd, ArH), 8.95 (1H, dd, ArH), 9.30 (1H, d, ArH), 10.75 (1H, t, NH). 54 C23H25N5O DCI+NH3, MH+388 (CDCL3), 2.40 (6H, s, 2xCH3), 2.70 (2H, t, CH2), 2.90 (6H, s, 2xCH3), 3.85 (2H, q, CH2), 7.40 (1H, d, ArH), 7.70 (1H, t, ArH), 7.90 (2H, dd, ArH), 8.40 (1H, dd, ArH), 8.53 (1H, d, ArH), 8.95 (1H, dd, ArH), 9.10 (1H, d, ArH), 10.85 (1H, br.NH) 55 C22H23N5O3 m/z438 (d4-MeOH)d 8.98(1H), 8.79(1H), 8.65(1H), 8.42(1H), 8.09(1H), 8.02-7.92(3H, m), 4.22 S (MH+) (2H, t), 3.70 (2H, t), 3.24 (3H, s), 3.18 (6H, s) No 2NH observed 56 C22H23N5O3S m/z 438(MH+) (d4-MeOH); 9.22 (1H, br.s), 8.91 (1H, dd, 7.3, 1.3), 8.49 (1H, d, 8.5), 8.19 (1H, d, 9.3), 8.13 (1H, dd, 7.4, 7.3), 8.07 (1H, d, 8.4), 7.92 (1H, d, 9.2), 7.60 (1H, dd, 8.4, 2.2), 4.29 (2H, t, 6.2), 3.71 (2H, t, 6.2 ), 3.25(3H, s), 3.18(6H, s) 57 C23H25N5O3S DCI+NH3MH+(452) CDCl3; 2.42 (6H, s), 2.73 (2H, t, J = 5.69Hz), 2.89 (6H, s), 3.85-3.92 (2H, m), 7.91 (1H, t, J = 7.91Hz), 8.02 (1H, t, J = 7.34Hz), 8.16 (1H, d, J = 9.83Hz), 8.41-8.51 (2H, m), 9.05 (1H, d, J = 5.56Hz), 9.09 (1H, d, J=9.84Hz), 9.85(1H, d, J=8.1Hz), 10.54(1H, broad peak) 58 C23H25N5O3S DCI+NH3MH+(452) CDCl3; 2.31(6H, s), 2.79(6H, s), 2.8-2.9(2H, m), 3.85-3.92(2H, m), 7.96(1H, t, J=7.19Hz), 8.05-8.15 (4H, m), 8.38 (1H, dd, J = 8.51Hz, J = 1.53Hz), 8.93 (1H, dd, J = 7.29Hz, 1.53Hz), 9.46 (1H, s), 10.32 (1H, wide peak) 59 C23H22N6O DCI+NH3, MNH4+, 416 (CDCl3), 2.45 (6H, s, 2xCH3), 2.80 (2H, t, CH2), 3.10 (2H, s, CH2), 3.90 (2H, q, CH2), 7.03 (1H, d, ArH), 7.75 (1H, t, ArH), 7.95 (2H, m, ArH), 8.12 (1H, d, ArH), 8.40 (1H, d, ArH), 8.80 (1H, d, ArH), 9.05 (1H, d, ArH), 10.95 (1H, br., NH). No 1 NH observed 60 C23H24N4O3 DCI/NH3m/z 405 (MH+, 100%) 400MHz, CDCl3.2.25(6H, s), 2.80(2H, t, J=6.1Hz), 3.82-3.87(2H, m), 4.06(3H, s), 4.10(3H, s), 6.78(1H, Broad peak), 7.19 (1H, d, J = 7.7Hz), 7.63-7.70 (2H, m), 7.85 (1H, d, J = 9.5Hz), 8.28 (1H, d, J = 9.4Hz), 8.45 (1H, d, J=9.4Hz), 8.95 (1H, d, J=8.2Hz). 61 C23H24N4O2 DCI+NH3, MH+389 CDCl3; 10.76(1H, br s); 8.97(2H, m); 8.52(1H, m); 8.40(1H, m); 7.95(2H, m); 7.75(1H, m); ); 4.05(3H, s); 3.93(1H, m); 3.70(1H, m); 3.08(1H, m); 2.45(6H, s)1.18(3H, dJ=6.51Hz) 62 C24H22F3N5O4 m/z 388(MH+) (d6-MeOH); 3.19 (6H, s), 3.71 (2H, t, J=6.0), 4.25 (2H, t, J=6.0), 8.01-8.16 (4H, m), 8.51 (1H, dd, J=8.7 and 1.4), 8.65 (1H, d, J=9.8), 8.82 (1H, dd, J=6.9 and 0.5), 9.27 (1H, d, J=8.0), no 3NH observed 63 C23H23ClN4O3 DCI/NH3m/e 439/441 (MH+, 100%/30%) 400MHz, CDCl32.25(6H, s), 2.62(2H, t, J=6.0Hz), 3.75-3.80(2H, m), 4.09(3H, s), 4.13(3H, s), 6.60(1H, Broad peak), 7.11 (1H, d, J = 8.7Hz), 7.73-7.78 (2H, m), 7.92 (1H, d, J = 9.5Hz), 8.26 (1H, d, 9.5Hz), 8.49 (1H , d, J=9.5Hz). 64 C21H21N5O3S DCI+NH3(MH+)424 CDCL3; 2.43(6H, s), 2.95-3.1(2H, m), 3.94-4.1(2H, m), 7.93(1H, t, J=7.88Hz), 8.06-8.1(3H, m), 8.35( 2H, t, J = 8.65Hz), 8.9 (1H, d, J = 5.87Hz), 9.59 (1H, s), 10.98 (1H, broad peak), no NH2 observed 65 C24H26N4O2 MH+at 403(100%) (d6-DMSO); 9.67 (br, 1H), 8.95 (d, 1H), 8.56 (d, 1H), 8.50 (d, 1H), 8.43 (d, 1H), 8.07 (m, 1H), 7.98 ( d, 1H), 7.86(m, 1H), 7.48(d, 1H), 4.07(s, 3H), 2.78(s, 2H), 2.30(s, 6H), 1.56(s, 6H). 66 C21H19N5O3 m/z 390 (CDCl3) 10.72 (1H, br, t), 9.98 (1H, d, 2.2), 8.80 (1H, dd, 6.6, 1.4), 8.56 (1H, dd, 8.5, 2.3), (MH+) 8.41 (1H, dd, 8.0, 1.4), 8.13 (1H, d, 9.3), 8.10-8.03 (2H, m), 7.98 (1H, dd, 7.3, 8.5), 4.26 (2H, q, 6.1), 3.59 (2H, q, 5.6), 2.92 (6H, d, 5.0) 67 C23H24N4O2 DCI/NH3m/z 389 (MH+, 100%) CDCl3, 400MHz, 2.42(6H, s), 2.76(2H, t, J=6.0Hz), 3.00(3H, s), 3.88-3.92(2H, m), 4.10(3H, s), 7.30(1H, d), 7.75-7.79 (1H, m), 7.82 (1H, d, J = 9.4Hz), 8.03 (1H, d, J = 9.7Hz), 8.55 (1H, d, J = 9.7Hz), 8.90 ( 1H, d, J=7.4Hz), 9.03(1H, d, J=8.0Hz), 10.95(1H, broad peak). 68 C21H20N4O3 DCI/NH3m/z 377 (MH+, 100%) 400MHz, d6-DMSO2.33(6H, s), 2.67(2H, t, J=5.7Hz), 3.75-3.80(2H, m), 7.32(1H, d, J=7.3Hz), 7.64(1H, d, J = 9.4Hz), 7.70-7.74 (1H, m), 7.85 (1H, d, J = 9.4Hz), 8.3 (1H, d, J = 9.4Hz), 8.40 (1H, d, J = 9.4 Hz), 8.78 (1H, d, J=8.1Hz), 10.55 (1H, broad peak), 11.4 (1H, broad peak). No 1 exchangeable proton observed 69 C24H24N4O4 MH+at433 (100%) d6-DMSO: 11.93 (1H, br.d), 8.94 (2H, m), 8.60 (1H, d, J=9.52Hz), 8.52 (1H, dd, J=8.6, 1.5Hz), 7.97 (2H, m), 7.88 (1H, t, J = 8.15Hz), 7.34 (1H, d, J = 7.95Hz), 5.61 (1H, m), 4.10 (3H, s), 3.96 (3H, s), 3.90- 3.70(2H, m), 3.04(6H, s) 70 C23H23ClN4O4 MH+at 419 d6-DMSO: 10.74 (1H, br.d), 9.01 (1H, d, J=8.19Hz), 8.69 (1H, d, J=7.06Hz), 8.55 (2H, m), 8.14 (1H, t, J=7.25Hz), 8.03(1H, d, J=9.49Hz), 7.86(1H, t, J=8.19Hz), 7.54(1H, d, 8.00Hz), 5.39(1H, m), 4.10(3H , s), 3.80-3.60 (2H, m), 2.95 (6H, br.s). 71 C24H26N4O2 m/z 403(MH+) (400MHz, CDCl3): 1.11 (3H, t, J=7.5), 1.78-1.89 (1H, m), 1.98-2.08 (1H, m), 2.36 (6H, s, NMe2), 2.59 (1H, dd, J=12.5 and 6.1), 2.82 (1H, dd, J=12.5 and 7.6), 4.10 (3H, s, OMe), 4.46-4.54 (1H, m, N-CH-), 7.27 (1H, d, J =9.0), 7.76(1H, t, J=8.1), 7.94-8.01(2H, m), 8.41(1H, d, J=8.5), 8.57(1H, d, J=9.4), 8.79(1H, d, J=8.2), 9.01 (1H, d, J=7.1), 10.92 (1H, d, J=8.3, NH) 72 C24H26N4O3 DCI/NH3m/e419 (MH+, 100%) 400MHz, CDCl31.55 (3H, d, J = 6.4Hz), 2.31 (6H, s), 2.39 (1H, dd, J = 12.2, 6.3Hz), 2.61 (1H, dd, J = 12.2, 8.6Hz) , 4.07(3H, s), 4.10(3H, s), 4.40-4.50(1H, m), 6.51(1H, broad peak d, J=6.1Hz), 7.20(1H, d, J=7.7Hz), 7.65-7.71(2H, m), 7.88(1H, d, J=9.6Hz), 8.28(1H, dJ=9.6Hz), 8.45(1H, d, J=9.7Hz), 8.98(1H, d, J =8.0Hz). 73 C22H21ClN4O2 DCI/NH3m/e 411/409 (MH+, 30, 100%) 400MHz, CDCl32.42(6H, s), 2.75(2H, t, J=5.9Hz), 3.86-3.90(2H, m), 4.08(3H, s), 7.25(1H, d, J=7.8Hz) , 7.72-7.77(1H, m), 7.89(1H, d, J=9.3Hz), 8.36(1H, d, J=2.5Hz), 8.56(1H, d, J=9.6Hz), 8.92(1H, d, J=2.5Hz), 8.95(1H, d, J=8.1Hz), 10.75(1H, broad peak). 74 C25H28N4O2 [M+H]+417 (400MHz, CDCl3) 10.90 (1H, d, br), 8.95 (1H, dd, J, 7.3, 1.4), 8.71 (1H, d, J 8.2), 8.48 (1H, d, J 9.6), 8.33 (1H , dd, J 8.6, 1.5), 7.90 (2H, m), 7.68 (1H, t, J 8.1), 7.18 (1H, d, 7.8), 4.49 (1H, m), 4.01 (3H, s), 2.83 (1H, dd, J 12.6, 9.1), 2.59 (1H, dd, J 12.6, 4.8), 2.28 (6H, s), 2.16 (1H, m) , 1.04 (3H, d, J 2.6), 1.03 (3H, d, J 2.6) 75 C23H24N4O3 DCI+NH3, MH+at 405 CDCl3: 11.05, (1H, d) 8.91, (1H, dd, J = 1.47Hz, 7.29Hz) 8.85, (1H, d, J = 8.17Hz) 8.50, (1H, d, J = 9.55Hz) 8.35, (1H, dd, J = 1.48, 8.47) 7.88, (2H, m) 7.71, (1H, t, J = 8.14) 7.20 (1H, s) 4.55, (1H, m) 4.05 (5H, m) 2.87, (2H, m)2.35, (6H, s). 76 C29H28N4O2 [M+H]+465 (400MHz, CDCl3) 11.0 (1H, d, br), 9.02 (1H, dd, J 7.2, 1.5), 8.55 (1H, d, J 9.4), 8.42 (1H, dd, 8.5, 1.6), 8.24 (1H , d, J 8.2), 7.99 (1H, d, J 8.3), 7.96 (1H, t, J 9.2), 7.55 (1H, t, J 8.1), 7.32 (2H, d, J 7.1), 7.22 (1H , d, J 8.0), 7.16 (2H, t, J 7.5), 7.04 (1H, t, J 7.3), 4.83 (1H, m), 4.09 (3H, s), 3.27 (1H, dd, J 13.7, 5.9), 3.21 (1H, dd, J 13.7, 5.5), 2.76 (1H, dd, J 12.4, 8.3), 2.60 (1H, dd, J 12.5, 6.1), 2.31 (6H, s) have an exchangeable Protons were not observed. 77 C23H24N4O2 DCI/NH3m/z 389 (MH+, 100%) 400MHz, CDCl31.53(3H, d, J=6.5Hz), 2.37(6H, s), 2.55(1H, dd, J=12.2, 6.2Hz), 2.84(1H, dd, J=12.2, 7.8Hz) , 4.10(3H, s), 4.55-4.65(1H, m), 7.28-7.30(1H, m), 7.75-7.80(1H, m), 7.95-8.02(2H, m), 8.42(1H, dd, J = 8.4, 1.5Hz), 8.57 (1H, d, J = 9.5Hz), 8.81 (1h, d, J = 8.2Hz), 9.02 (1H, dd, J = 7.3, 1.6Hz), 10.9 (1H, d, J=7.2Hz). 78 C23H24N4O2 m/z 389(MH+) (400MHz, CDCl3): 1.53 (3H, d, J=6.5, Me), 2.38 (6H, s, NMe2), 2.51-2.60 (1H, m), 2.82-2.90 (1H, m), 4.11 (3H, s, MeO), 4.56-4.65 (1H, m, CH), 7.29 (1H, d, J=8.2), 7.78 (1H, t, J=8.1), 7.96-8.02 (2H, m), 8.43 (1H , d, J=8.4), 8.58 (1H, d, J=9.5), 8.82 (1H, d, J=8.4), 9.03 (1H, d, J=7.3) 79 C22H21N5O3 DCI+NH3, MH+, 404 400MHz(CDCL3), 1.50(3H,d,), 2.35(6H,s), 2.55(1H,dd), 2.80(1H,dd), 4.60(1H,m), 7.90(1H,t), 8.10( 1H, t,), 8.25 (1H, d,), 8.45 (2H, m.,), 8.70 (1H, d), 9.05 (1H, dd), 9.65 (1H, d,), 10.45 (1H, d , NH). 80 C22H21N5O3 DCI+NH3, MH+404 400MHz(CDCL3), 1.60(3H,d.), 2.30(6H,s), 2.65(1H,m), 3.05(1H,m), 4.70(1H,m), 8.05(1H,dd), 8.70( 1H, d), 8.75 (1H, d), 8.45 (2H, m,), 8.70 (1H, d), 9.10 (1H, d,), 10.00 (1H, d), 10.45 (1H, br).- 81 C23H24N4O3 DCI+m/z405(MH+, 100%) 400MHz, 10.97 (1H, br) in CDCl3; 8.91 (1H, dd, J=7.2 &1.5Hz); 8.76 (1H, d, J=8.2Hz); 8.41 (1H, d, J=9.5Hz); 8.35 (1H, dd, J=8.6 &1.5Hz); 7.90(1H, m); 7.86(1H, d, J=9.5Hz); 7.72(1H, t, J=8.0Hz); 7.21(1H, d, J=8.0Hz); 5.32(1H,br); 4.63(1H,m); 4.17(2H,m); 4.06(3H,s); 2.96(1H,dd, J=12.4 &6.0Hz); 2.88( 1H,dd,J=12.4 &7.4Hz); 2.39(6H,s). 82 C23H25N5O2 MH+at 404 CDCl3: 11.92, (1H, m) 11.12, (1H, m) 8.80, (1H, d, J=8.33Hz) 8.40, (1H, d, J=9.53Hz) 8.12, (1H, d, J = 9.62Hz) 7.90, (1H, d, J = 9.45Hz) 7.70, (1H, t, J = 8.09Hz) 7.56, (1H, d, J = 9.59Hz) 7.20, (1H , d, J=7.83Hz) 4.10, (3H, s) 3.83, (2H, q, J=6.16Hz) 3.18, (3H, d, J=5.11Hz) 2.77, (2H, t, J=6.51Hz )2.40, (6H,s) 83 C23H24N4O3 DCI/NH3MH+405(100%) 400MHz, CDCl31.52 (3H, d, J = 6.6Hz), 2.37 (6H, s), 2.56 (1H, dd, J = 12.4, 6.0Hz), 2.85 (1H, dd, J = 12.4, 8.1Hz) , 4.10(3H, s), 4.52-4.62(1H, m), 7.25(1H, d, J=7.8Hz), 7.57(1H, d, J=9.4Hz), 7.75-7.78(1H, m), 7.96 (1H, d, J = 9.5Hz), 8.21 (1H, d, J = 9.4Hz), 8.49 (1H, d, J = 9.3Hz), 8.68 (1H, d, J = 9.2Hz), 11.70 ( 1H, d, broad peak), 16.2 (1H, s). 84 C24H26N4O3 m/z CI+419 (MH+, 100%); 305 (55%) 400MHz in CDCl3 11.21(1H, br); 9.27(1H, d, J=8.2Hz); 9.02(1H, dd, J=7.3 &1.5Hz); 8.53(1H, d, J=9.6Hz); 8.42( 1H, dd, J=8.6 &1.5Hz); 7.99-7.91(2H, m); 7.75(1H, t, J=8.2Hz); 7.35(1H, d, J=8.0Hz); 4.43(2H, m ); 4.06(3H, s); 3.48(1H, s); 3.06(1H, dd, J=13.1 &5.2Hz); 2.88(1H, dd, J=13.1 &4.8Hz); 2.36(6H, s) ;1.37(3H,d,J=6.4Hz). 85 C22H22N4O3 DCI/NH3MH+391(100%) 400MHz, CDCl3 2.42(6H, s), 2.78(2H, t, J=6.0Hz), 3.86-3.90(2H, m), 4.10(3H, s), 7.25(1H, d, J=7.8Hz), 7.57(1H, d, J=9.4Hz), 7.70-7.75(1H, m), 7.95(1H, d, J=9.4Hz), 8.22(1H, d, J=9.4Hz), 8.51(1H, d , J=9.6Hz0, 8.81(1H, d, J=8.2Hz), 11.65(1H, br), 16.17(1H, s). 86 C25H26N4O2 DCI/NH3m/e 414.4 (MH+, 100%) 400MHz, CDCl31.35(2H, m), 1.45(4H, m), 2.47(4H, m), 2.7(2H, t), 3.7(2H, m), 4.07(3H, s), 7.5(1H, d) 7.83(1H,m), 7.95(1H,d), 8.07(1H,m), 8.43(1H,m), 8.5(1H,m), 8.62(1H,m), 8.73(1H,m) , 10.05 (1H, broad peak) 87 C24H26N4O3 [M+H]+419 (400MHz, CDCl3) 11.05 (1H, d), 8.92 (1H, dd, J 7.2, 1.5), 8.53 (1H, d, J 8.2), 8.48 (1H, d, J9.5), 8.35 (1H, dd , J 8.5, 1.5), 7.94-7.86 (2H, m), 7.69 (1H, t, J 8.1), 7.19 (1H, d, J 7.7), 4.59 (1H, q, J 7.1), 4.02 (3H, s), 3.80-3.67 (2H, m), 3.44 (1H, dd, J12.2, 6.0), 2.63 (1H, dd, J12.2, 2.1), 2.35 (6H, s), 2.10 (1H, m), 1.99(1H, m), with 1 exchangeable proton not observed 88 C22H23N5O2 MH+at 390 CDCl3-11.75, (m, 1H) 8.80, (d, 1H, J=8.24Hz), 8.43, (1H, J=9.29Hz) 8.01, (d, 1H, J=9.32Hz) 7.91, (d, 1H , J=9.48Hz) 7.72, (t, 1H, J=8.10Hz) 7.23, (d, 1H, J=9.34Hz) 7.20, (d, 1H, J=7.74Hz) 4.09, (s, 3H) 3.87 , (q, 2H, J=6.10) 2.70, (t, 2H J=6.43Hz) 2.42, (s, 6H).NH2 not observed. 89 C24H24N4O3 DCI/NH3m/e 417.2(MH+) 400MHz, CDCl32.62(4H, t), 2.9(2H, t), 3.65(4H, t), 3.93(2H, m), 4.20(3H, s), 7.25(1H, m), 7.7(1H, t), 8.00(2H, m), 8.45(1H, m), 8.60(1H, d), 8, 75(1H, d), 9.0(1H, d), 10.9(1H, broad peak) 90 C24H24N4O2 DCI/NH3 400MHz, CDCl3 m/e 401(MH+) 1.82(4H,t), 2.75(4H,t), 3.2(2H,t), 3.90-3.99(2H,m), 4.11(3H,3), 7.26-7.31(1H,m), 7.78(1H, t), 7.95-7.98 (1H, m), 7.98-8.03 (1H, m), 8.4-8.46 (1H, m), 8.55-8.61 (1H, m), 8.78-8.83 (1H, m), 8.98-9.3 (1H, m) NH not observed 91 C24H26N4O4 DCI/NH3m/e 435.3(MH+) 400MHz, CDCl32.75(4H, t), 3.00(2H, t), 3.56(4H, t), 3.80-3.89(2H, m), 4.40(3H, s), 7.16-7.22(1H, m), 7.69-7.75(1H, m), 7.85-7.89(1H, m), 7.89-7.93(1H, m), 8.33-8.39(1H, m), 8.47-8.51(1H, m), 8.51-8, 55 (1H, m), 8.89-8.95 (1H, m), 11.29 (1H, broad peak), 2xOH not observed 92 C24H26N4O2 DCI/NH3m/e 403.3(MH+) 400MHz, DMSO0.99(6H, t), 2.65(4H, q), 2.79(2H, t), 3.65-3.72(2H, m), 4.07(3H, s), 7.49-7.53(1H, m), 7.8-7.87(1H,m), 7.98(1H,d), 8.04-8.11(1H,m), 8.45(1H,m), 8.51(1H,m), 8.67(1H,m), 8.89(1H, m), 10.15-10.25 (1H, broad peak). 93 C23H24N4O2S ESI+ve m/e421 (MH+, 100%) 400MHz, CDCl32.42(6H, s), 2.70(3H, s), 2.75(2H, t, J=6.0Hz), 3.85-3.90(2H, m), 4.10(3H, s), 7.24(1H, d, J = 8.0Hz), 7.72-7.77 (1H, m), 7.91 (1H, d, J = 9.5Hz), 8.00 (1H, d, J = 2.4Hz), 8.55 (1H, d, J = 9.7 Hz), 8.85(1H, d, J=2.4Hz), 8.99(1H, d, J=8.0Hz), 10.85(1H, broad peak). 94 C23H24N4O3 ESI+ve m/e405 (MH+, 100%) CDCl3, 400MHz2.42(6H, s), 2.75(2H, t, J=6.0Hz), 3.85-3.90(2H, m), 4.07(3H, s), 4.10(3H, s), 7.22(1H, d, J = 8.0Hz), 7.64 (1H, d, J = 3.0Hz), 7.72-7.77 (1H, m), 7.91 (1H, d, J = 9.4Hz), 8.55 (1H, d, J = 9.6 Hz), 8.70(1H, d, J=3.0Hz), 8.99(1H, d, J=8.0Hz), 10.92(1H, broad peak). 95 C24H26N4O4 DCI/NH3m/e 435 (MH+, 100%) 400MHz, CDCl32.32 (6H, s), 2.65 (1H, dd, J = 12.3, 6.7Hz), 2.75 (1H, dd, J = 12.3, 8.1Hz), 3.98 (1H, dd, J = 11.3, 4.1 Hz), 4.05 (3H, s), 4.09 (3H, s), 1H on 2 methoxy peaks, 4.41-4.48 (1H, m), 6.91 (1H, d, J=5.3Hz), 7.20 ( 1H, d, J = 7.9Hz), 7.67-7.71 (2H, m), 7.86 (1H, d, J = 9.6Hz), 8.29 (1H, d, J = 9.4Hz), 8.45 (1H, d, J = 9.7Hz), 8.91 (1H, d, J = 8.2Hz). No 1 exchangeable proton observed 96 C21H20N4O2 DCI/NH3m/e 361.0(MH+) 400MHz, CDCl32.64(3H, s), 3.12(2H, t), 3.90-3.96(2H, m), 4.11(3H, s), 7.29(1H, s), 7.63-7.71(1H, m), 7.92-7.95(1H, m), 7.95-8.01(1H, m), 8.38-8.45(1H, m), 8.55-8.61(1H, m), 8.89-8.95(1H, m), 8.95-9.02(1H , m), 11.08 (1H, broad peak). 97 C23H24N4O4 DCI/NH3m/z 421 400MHz, CDCl32.40(6H, s), 2.92(1H, dd, J=12.4, 7.6Hz), 2.99(1H, dd, J=12.4, 5.9Hz), 4.07(3H, s), 4.18- (MH+, 100%) 4.20(2H, m), 4.58-4.68(1H, m), 7.21(1H, d, J=7.9Hz), 7.53(1H, d, J=9.5Hz), 7.70-7.73(1H, m), 7.86 (1H, d, J = 9.4Hz), 8.15 (1H, d, J = 9.4Hz), 8.37 (1H, d, J = 9.3Hz), 8.65 (1H, d, J = 8.3Hz), 11.70 (1H , d), 15.82 (1H, s) no 1 exchangeable proton observed 98 C25H28N4O2 [M+H]+417 (400MHz, CDCl3) 11.0 (1H, br, d), 9.03 (1H, dd, J 7.3, 1.6), 8.80 (1H, d, J 8.3), 8.56 (1H, d, J 9.5), 8.41 (1H, dd, J 8.5, 1.5), 7.97 (2H, m), 7.75 (1H, t, J 8.1), 7.26 (1H, d, J 7.7), 4.56 (1H, m), 4.09 (3H, s), 2.89 (1H, dd, J 12.6, 9.0), 2.64 (1H, dd, J 12.6, 5.1), 2.34 (6H, s), 2.22 (1H, m), 1.12 (3H, d, J 2.5), 1.10 (3H , d, J 2.4) 99 C23H22N4O2 DCI/NH3m/z 387 (MH+, 100%) CDCl3/400MHz, 2.02-2.10(1H, m), 2.40-2.46(1H, m), 2.52-2.61(1H, m), 2.58(3H, s), 2.75-2.79(1H, m), 2.99-3.02 (1H, m), 3.13-3.18 (1H, m), 4.10 (3H, s), 4.90-4.98 (1H, m), 7.26 (1H, d), 7.75-7.80 (1H, m), 7.95-8.00 (2H, m), 8.42 (1H, dd, J = 8.4, 1.5Hz), 8.58 (1H, d, J = 9.6Hz), 8.99-9.03 (2H, m), 11.32 (1H, d). 100 C25H29N5O2 MH+432 (400MHz, CDCl3); 2.25(6H, s), 2.32-2.38(1H, m), 2.52(6H, s), 2.56-2.62(1H, m), 3.00-3.09(1H, m), 3.65-3.74 (1H, m), 4.01-4.09 (1H, m), 4.11 (3H, s), 7.28 (1H, d, J=7.7), 7.78 (1H, t, J=8.0), 7.96-8.01 (2H, m), 8.42 (1H, d, J=8.5), 8.58 (1H, d, J=9.6), 8.98-9.03 (2H, m)

Claims (21)

1. one kind is compound or its pharmacologically acceptable salts of benzo [a] azophenlyene-11-amide derivatives of formula (I):

R in the formula ¹-R ³Be hydrogen, R ⁴The C that is selected from hydroxyl and does not replace or replaced by cyano group ₁-C ₆Alkoxyl group;

R ⁵-R ⁷Identical or different, be selected from hydrogen, hydroxyl and C separately ₁-C ₆Alkoxyl group;

Q is C ₁-C ₆Alkylidene group, its C for not replacing or do not replaced or being replaced by hydroxyl ₁-C ₆Alkyl replaces;

R ⁸And R ⁹Identical or different, hydrogen or C respectively do for oneself ₁-C ₆Alkyl; With

R ¹⁰Be hydrogen or C ₁-C ₆Alkyl.

2. the compound of claim 1 or its pharmacologically acceptable salts, wherein said benzo [a] azophenlyene-11-amide derivatives has the structure of formula (Ia):

R in the formula ¹-R ⁹Definition as claimed in claim 1;

P is 1,2 or 3; With

R ¹³Be (i) hydrogen, the C that does not perhaps (ii) replace or replaced by hydroxyl ₁-C ₆Alkyl.

3. claim 1 or 2 compound or its pharmacologically acceptable salts, wherein R ⁴Be C ₁-C ₆Alkoxyl group or hydroxyl.

4. claim 1 or 2 compound or its pharmacologically acceptable salts, wherein R ⁷Be hydroxyl.

5. the compound of claim 1 or its pharmacologically acceptable salts, it is selected from:

4-methoxyl group-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-ethyl)-acid amides

4-hydroxyl-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-ethyl)-acylamino hydrogen bromate

4-cyano group methoxyl group-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-ethyl)-acid amides

4,10-dihydroxyl-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-ethyl)-acid amides

4-methoxyl group-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-1-methylol-ethyl)-acid amides

4-methoxyl group-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-1-(R)-methyl-ethyl)-acid amides

4-methoxyl group-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-1-(S)-methylol-ethyl)-acid amides

4-methoxyl group-benzo [a] azophenlyene-11-formic acid (1-dimethylamino methyl-2-hydroxyl-propyl group)-acid amides

4,9-dimethoxy-benzo [a] azophenlyene-11-formic acid (2-dimethylamino-ethyl)-acid amides

(R)-4-methoxyl group-benzo [a] azophenlyene-11-formic acid (1-dimethylamino methyl-2-methyl-propyl group)-acid amides.

6. medicinal compositions, it comprises as each compound or its pharmacy acceptable salt and pharmaceutically acceptable carrier or thinner in the claim 1 to 5 of activeconstituents.

7. method for preparing compound or its pharmacy acceptable salt of claim 1, described method comprises:

(a) amine of employing formula (III):

Q, R in the formula ⁸And R ⁹As definition in the claim 1, the activated derivatives of the compound of processing formula (II):

R in the formula ¹To R ⁷As definition in the claim 1; Or

(b) in organic solvent, or do not use solvent, under the elevated temperature condition, adopt the compound of the compound treatment formula (IV) of formula (III) as defined above:

R in the formula ¹To R ⁷As definition in the claim 1, R ¹¹Be C ₁-C ₆Alkyl, C ₃-C ₆Cycloalkyl, benzyl or phenyl; With

(c) if desired, by the chemical modification method of routine, benzo [a] azophenlyene-11-amide derivatives of gained is converted into another kind of this analog derivative, and/or benzo [a] azophenlyene-11-amide derivatives is converted into its pharmacy acceptable salt.

8. the compound of a formula (II):

R in the formula ¹To R ⁷Definition as claimed in claim 1, or its salt or ester.

9. method for preparing formula (II) compound of claim 8, described method comprises:

(a) in organic solvent, the phenylformic acid of employing formula (VI):

Or its ester or salt, wherein R ⁵, R ⁶And R ⁷As definition in the claim 1,

Handle 1 of formula V, the 2-naphthoquinones:

R in the formula ¹To R ⁴As definition in the claim 1.

10. the method for claim 9, it carries out in the presence of 1 to 5 normal mineral acid.

11. the purposes of each compound in the medicine for preparing the inhibitor that is used as topoisomerase I and/or topoisomerase II in the claim 1 to 5.

12. the purposes of claim 11, wherein said medicine are the inhibitor as topoisomerase I.

13. the purposes of claim 11, wherein said medicine are the inhibitor as topoisomerase II.

14. the purposes of claim 11, wherein said medicine are the combined depressants as topoisomerase I and topoisomerase II.

15. any one purposes of claim 11-14, wherein said medicine is used for the treatment of tumour.

16. the purposes of claim 15, wherein said tumour shows multidrug resistance.

17. the purposes of claim 16, wherein said multidrug resistance are the multidrug resistance that is situated between between P-glycoprotein.

18. the purposes of claim 16, wherein said multidrug resistance are the multidrug resistance that is situated between between MRP.

19. the purposes of claim 16, wherein said multidrug resistance are the atypia multidrug resistance.

20. the purposes of claim 11, wherein said medicine when treatment suffers from the mankind of tumour or animal with the oral way administration.

21. the purposes of claim 11, wherein said medicine when treatment suffers from the mankind of tumour or animal with the administration of non-enteron aisle mode.