HK1138281B - Minor groove binders - Google Patents

Abstract

There is provided compounds of formula (I), wherein R1, R11, R12, Qa, X, Qb, Qc, A and D have meanings given in the description, or a pharmaceutically acceptable salt or solvate thereof, which compound, salt or solvate binds to the minor groove of DNA.

Description

Minor groove binders

Technical Field

The present invention relates to synthetic compounds having affinity for nucleic acids, particularly compounds that bind to the minor groove of DNA.

Background

The listing or discussion of a prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is prior art or common general knowledge.

Due to its fundamental role in molecular biological processes, DNA represents an important target for drug action. Compounds that recognize defined sequences of DNA have a variety of potential uses, such as modulating gene expression.

The outer surface of double-helical DNA has two grooves, called major and minor. These channels all contain chemical information represented by the arrangement of hydrogen-binding donors and acceptors, electrostatic charges, dipoles, hydrophobic regions, and the like.

Depending on the number of potential hydrogen-binding contacts, the major groove contains approximately twice the amount of information as the minor groove. In this regard, the major groove is the preferred recognition site for cellular proteins, such as regulatory proteins, promoters and repressors.

In contrast, the minor groove is generally (and with some exceptions) relatively unoccupied. This weakness of the minor groove makes it particularly useful for targeting compounds that bind to DNA. Indeed, perhaps for this reason, the minor groove is the binding site for certain naturally occurring antibiotics (e.g., fusiform and distamycin).

Fusins and distamycin are oligopeptides based on pyrrole amino acid monomers. These compounds are all represented by 10^-5The dissociation constant of M binds to DNA. They also tend to bind to the AT rich region of DNA. Despite their intrinsic biological activity, fusidic and distamycin also have many limitations, including toxicity, moderate affinity and limited selectivity. Some workers have therefore prepared synthetic analogues of fusidic and distamycin to overcome these disadvantages. Many of these compounds are reviewed by Sondhi et al (curr. Med. chem.4, 313(1997)), Reddy et al (Pharmacology)&Therapeutics 84, 1(1999)), Wemmer (Biopolymers 52, 197(2001)), and Dervan (bioorg.med.chem.9, 2215 (2001)).

Compounds intended to bind to regions of DNA containing GC base pairs are described below, for example: Anti-Cancer Drug Design 5, 3 (1990); proc.natl.acad.sci USA 89, 7586 (1992); biochemistry 32, 4237 (1993); science 266, 647 (1994); Anti-Cancer Drug Design 10, 155 (1995); med, chem.8, 985 (2000); and moi. biol.34, 357 (2000). Other various analogs of fusidic and distamycin are described below: am chem soc.114(15), 5911 (1992); biochemistry31, 8349 (1992); bioconjugate chem.5, 475 (1994); biochem. biophysis. res. commun.222, 764 (1996); med chem.43, 3257 (2000); and Tetrahedron 56, 5225 (2000). In addition, the use of certain fusidic and distamycin analogs as antibacterial, antiviral and/or antitumor agents is described below: molecular pharmacology 54, 280(1998), bioorg.med.chem.lett.6(18), 2169(1996), j.med.chem.45, 805(2002), bioorg.med.chem.12, 2007(2002), international patent applications WO 97/28123, WO 98/21202, WO 01/74898 and WO 02/00650, and U.S. patent nos. 4,912,199, 5,273,991, 5,637,621, 5,698,674 and 5,753,629. Methods for synthesizing fusidic and distamycin analogs are described in us 6,090,947.

Cellular uptake of distamycin analogs is described in bioorg.med.chem.lett.11, 769 (2001).

Compounds intended for further binding to DNA are described in U.S. Pat. No. 6,143,901, which discloses oligomers between 6 and 30 heterocyclic groups, where the group linking the heterocyclic rings may be methyleneamino, amino, thioamino, iminydyl or ethenylene. Amino groups (and heterogeneous analogs thereof) are described as preferred linking groups. If these groups are present, there is no preference in the number or position of the linking groups to the vinylidene groups in U.S. Pat. No. 6,143,901. Furthermore, there is no indication that vinylidene containing compounds may have any advantage over compounds containing other linkages such as amino groups.

Analogues of distamycin are described in Tet.Lett.37(43), 7801-7804(1996), wherein the amino group linking the two pyrrolyl groups of the oligomeric pyrrole compound is replaced by a diketone or alkenylene linker. The resulting compounds are described as having significantly reduced binding affinity for DNA compared to analogous amino-linked compounds.

Minor groove binding compounds containing an acrylamide-type linker between the pyrrole group at position 2 and the terminal base are described below: J.Am.chem.Soc.122, 1602-.

Additional minor groove binding compounds containing an acrylamide-type linker between the pyrrole group at position 3 and the terminal base are described in J.Med.chem.47, 2133-2156 (2004).

None of the above documents discloses or suggests compounds with DNA affinity, wherein the compounds comprise oligomers of cyclic groups wherein an alkenylene group directly links one cyclic group to an adjacent cyclic group at the "amino" end and at least one of the cyclic groups linked via an alkenylene moiety is not a pyrrole (or 5-membered heterocycle).

Disclosure of Invention

According to the present invention there is provided a compound of formula I, or a pharmaceutically acceptable derivative thereof,

wherein the wavy line indicates optional cis or trans stereochemistry;

R¹representative H, R^1aC(O)-NH-、NO₂or-N (R)^2a)R^2b；

R^1aRepresents aryl (which is represented by the following groups selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy groups are optionally substituted),

aromatic or partially aromatic C_13-14Tricyclic carbocycle (which is substituted at the last group by a group selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4Alkoxy radicalOptionally substituted with one or more substituents of (a), and this latter group, if partially aromatic, is optionally substituted in the nonaromatic moiety by one or two oxygen-containing groups) or

C_1-12Alkyl (which is substituted by a group selected from halo or aryl (which is substituted by a group selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy being optionally substituted) is optionally substituted and/or blocked;

a represents 1, 2, 3 or 4;

a represents C_2-6Alkylene or A¹-C(O)N(H)-A²Wherein A is²To the D group;

A¹represents C_1-4An alkylene group;

A²represents C_2-5An alkylene group;

d represents Het¹、-N(R^3a)R^3b、-C(＝NR^3c)N(R^3d)R^3eor-N (R)^3f)C(＝NR^3g)N(H)R^3h；

Het¹Represents a4 to 12 membered heterocyclic group containing at least one N atom and optionally one or more heteroatoms selected from N, O and S, consisting of ═ O, OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy are optionally substituted;

R^3aand R^3bIndependent representation H, C_1-6Alkyl or Het²；

R^3cTo R^3hIndependently represent H or C_1-6An alkyl group;

Het²independently represents a 4-to 12-membered heterocyclic group containing one or more heteroatoms selected from N, O or S, the heterocyclic substituent being substituted by a substituent selected fromO, OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy are optionally substituted;

as used herein, each Q^aTo Q^cEach independently represents naphthalene (substituted by one or more substituents selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy optionally substituted), Het³Or a structural fragment of the formula Ia, Ib, Ic, Id, Ie or If,

wherein the dashed lines indicate the attachment positions of the fragments;

R⁴represents H or C_1-6An alkyl group;

R⁵represents C_1-12An alkyl group;

R⁶、R⁷、R⁸and R⁹Independently represent H or C_1-12An alkyl group;

R¹⁰each independently represents OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl, -S-C_1-4Alkyl or C_1-4An alkoxy group;

b represents 0, 1, 2 or 3;

G¹and G²Independently represent CH or N, or G²Optionally represents C-R¹⁰；

L represents O or S;

Het³represents a 9 or 10 membered bicyclic heterocyclic group containing one or more heteroatoms selected from N, O or S, consisting of ═ O, halo, cyano, nitro, N (R)^2a)R^2b、Het^a、C_1-4Alkyl OR OR^aOptionally substituted with one or more substituents of (a);

R^arepresentative H, C_1-4Alkyl, aryl (which is substituted at the latter group by a member selected from the group consisting of OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy being optionally substituted) or Het^b；

Het^aAnd Het^bIndependently represents a 4-to 12-membered heterocyclic group containing one or more heteroatoms selected from N, O or S, which heterocyclic group is represented by the general formula (I) selected from ═ O, OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy are optionally substituted;

when used herein, R¹¹And R¹²At each place represent H, C independently_1-6Alkyl or aryl (which is substituted in the latter group by a substituent selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy are optionally substituted);

when used herein, R^2aAnd R^2bIndependently represent H or C_1-4Alkyl, or R^2arepresents-C (O) R¹³；

R¹³Represents H or C_1-4An alkyl group;

and unless otherwise specified, the alkyl portion of the alkyl, hydrocarbylene, alkenylene, cycloalkyne, phenylene and naphthylene, and alkoxy groups, may be substituted with one or more halogen atoms;

with the proviso that the compound contains at least one Q^aOr Q^bA group other than G¹A structural fragment of the general formula Ia representing CH,

these compounds are hereinafter referred to as "compounds of the invention".

Unless otherwise specified, the alkyl and alkoxy groups defined herein may be straight chain or, when there is a sufficient number (i.e. at least 3) of carbon atoms, branched and/or cyclic. In addition, when there are a sufficient number (i.e., at least 4) of carbon atoms, for example, alkyl and alkoxy groups, the groups may also be partially cyclic/acyclic. Such alkyl and alkoxy groups may also be saturated or, when there are a sufficient number (i.e. at least 2) of carbon atoms, unsaturated and/or interrupted by one or more oxygen and/or sulfur atoms. Unless otherwise specified, the alkyl and alkoxy groups may also be substituted by one or more halogen atoms, in particular fluorine atoms.

Unless otherwise specified, the hydrocarbylene group as defined herein may be straight chain or, when there is a sufficient number (i.e. at least 2) of carbon atoms, branched. Such alkylene chains may also be saturated or, when there are a sufficient number (i.e. at least 2) of carbon atoms, unsaturated and/or interrupted by one or more oxygen and/or sulfur atoms. Unless otherwise specified, the alkylene group may also be substituted by one or more halogen atoms.

The term "aryl", as used herein, includes C_6-10Aryl groups such as phenyl, naphthyl, and the like. When substituted, the aryl group is preferably substituted between 1 and 3 substituents.

The term "heterocyclic group" as used herein includes 4 to 12 membered (e.g. 5 to 10 membered) heterocyclic groups containing one or more heteroatoms selected from N, O or S. This term thus includes mono-or bicyclic, and may be saturated, partially unsaturated, aromatic, or, where appropriate, partially aromatic, such groups. Preferred heterocyclic groups include aromatic or partially aromatic groups such as pyrrolyl, imidazolyl, thiazolyl, oxazolyl, benzoxazolyl, furanyl, thienyl, pyridyl and coumarinyl. Particularly preferred heterocyclic groups include pyrrolyl, imidazolyl, thiazolyl and oxazolyl.

"substituted on the heterocyclic moiety" means that each of the important branches, cyclic or partially cyclic C_3-5Alkyl radicals being in each caseA direct substituent on the heterocyclic ring of a heterocyclic monomer having such a group (whether attached to the ring through a heteroatom or otherwise).

The term "aromatic or partially aromatic C_13-14Tricyclic carbocycles ", when used herein, include fluorenyl, anthracenyl, 9, 10-dihydroanthracenyl, phenanthrenyl, 9, 10-dihydrophenanthrenyl, and the like.

The term "halo", as used herein, includes fluoro, chloro, bromo and iodo.

Mentioned Het (Het)¹To Het³，Het^aAnd Het^b) Groups include those containing 1 to 4 heteroatoms (selected from oxygen, nitrogen and/or sulfur groups) and wherein the total number of atoms in the ring system is between 5 and 12. Het (Het)¹To Het³，Het^aAnd Het^b) The groups may be fully saturated, partially unsaturated, wholly aromatic, partially aromatic and/or bicyclic in nature. Heterocyclic groups mentioned include 1-azabicyclo [2.2.2]Octaalkyl, benzimidazolyl, benzo [ c ]]Isoxazolidinyl, benzisoxazolyl, benzodioxanyl, benzodioxolyl, benzofuranyl, benzofurazanyl, benzomorpholinyl, 2,1, 3-benzoxadiazolyl, benzoxazolinyl, benzoxazolyl, benzopyrazolyl, benzo [ e ] alkyl]Pyrimidinyl, 2,1, 3-benzothiazolyl, benzothiophenyl, benzotriazolyl, benzoxazolyl, heteroalkyl, chromenyl, cinnolinyl, coumarinyl, 2, 3-dihydrobenzimidazolyl, 2, 3-dihydrobenzo [ b]Furyl, 1, 3-dihydrobenzo [ c ]]Furyl, 1, 3-dihydro-2, 1-phenylisoxazolyl, 2, 3-dihydropyrrolo [2, 3-b ]]Pyridyl, dioxinyl, furyl, hexahydropyrimidyl, hydantoinyl, imidazole, imidazo [1, 2-a ]]Pyridyl, imidazo [2, 3-b ]]Thiazolyl, indolyl, isoquinolinyl, isoxazolidinyl, isoxazolyl, maleimido, morpholinyl, oxadiazolyl, 1, 2-or 1, 3-oxazinyl, oxazolyl, phthalazinyl, piperazinyl, piperidinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrrolidonyl, pyrrolylAlkyl, pyrrolinyl, pyrrolo [2, 3-b ]]Pyrimidinyl, pyrrolo [5, 1-b ]]Pyridyl, pyrrolo [2, 3-c ]]Pyridyl, pyrrolyl, quinazolinyl, quinolinyl, dioxothienylalkyl, 3-dioxothienylalkenyl, 4, 5,6, 7-tetrahydrobenzimidazolyl, 4, 5,6, 7-tetrahydrobenzopyrazolyl, 5,6, 7, 8-tetrahydrobenzo [ e ] l]Pyrimidinyl, tetrahydrofuryl, pyranyl, 3, 4, 5, 6-tetrahydropyridyl, 1, 2, 3, 4-tetrahydropyrimidinyl, 3, 4, 5, 6-tetrahydropyrimidinyl, thiadiazolyl, thiazolidinyl, thiazolyl, thienyl, thieno [5, 1-c ]]Pyridyl, thiochromanyl, triazolyl, 1, 3, 4-triazolo [2, 3-b ]]Pyrimidinyl, xanthenyl, and the like.

Mentioned Het¹The meaning of (a) includes the pyrrolodin-1-yl or, in particular, morpholin-4-yl.

Mentioned Het³The meaning of (A) includes isoquinolinyl or, in particular, quinolyl (e.g. when Q is^aRepresents Het³When mentioned, Het³The meanings of (A) include quinolin-2-yl or quinolin-3-yl, wherein the numbering of the position of the attachment is relative to-C (R)¹¹) Determined by the connection location).

Mentioned Het^bIncluded within the meaning of (a) include benzotriazolyl (e.g., benzo-1-yl).

Pharmaceutically acceptable derivatives include salts and solvates. The salts mentioned include acid addition salts.

The compounds of formula I exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention.

The compounds of formula I may also contain one or more asymmetric carbon atoms and may therefore exhibit optical and/or diastereoisomerism. The diastereomers may be separated using conventional techniques, such as chromatography or fractional crystallization. The various stereoisomers may be separated by separation of the racemates or other mixtures of the compounds using conventional techniques, such as fractional crystallization or high performance liquid chromatography. Alternatively, the desired optical isomer may be isolated by reaction of appropriate optically active starting materials under conditions which do not cause racemisation or epimerisation, or by derivatization, e.g. with a chiral acid followed by separation of the diastereomeric ester by conventional methods (e.g. high performance liquid chromatography, chromatography over silica). All stereoisomers are included within the scope of the present invention.

The compounds of the invention may be provided in a bioavailable form. As used herein, the term "bioavailable" includes compounds that, when used, are in a form that can interact with a biological system to provide a measurable therapeutic response. This term may thus be understood to include compounds that are provided to the DNA in a form and/or level sufficient to provide a measurable desired or required therapeutic response.

The bioavailability of a compound is predicted by a variety of methods well known to those skilled in the art, including measurement of the partition coefficient of the compound between water (e.g., at a pH between 5 and 9) and an organic, water-insoluble solvent (e.g., octanol), which can be used to predict the behavior of the compound in question in human tissue (see, e.g., j. med. chem.43, 3257-3266 (2000)).

Bioavailability is achieved by providing a compound of the invention in a form (e.g., pharmaceutical formulation) that is presented to the DNA at an appropriate concentration to provide a measurable therapeutic response. Bioavailability can also be achieved by altering the physicochemical properties of the active center, such as by increasing water solubility (e.g., by introducing additional basic groups, as described in j.med.chem.43, 3257-3266(2000)) using techniques well known to those skilled in the art.

The compounds of the invention may have a high affinity for at least one DNA sequence. As used herein, the term "having high affinity for at least one DNA sequence" includes having a molecular weight of at least 10 when bound to the minor groove of at least one DNA oligomer or polymer^-5A dissociation constant of less than M, preferably 10^-6M (e.g. 10)^-7M) below, in particular at 10^-8Compounds having M or less. In this regard, the dissociation constant may be atIt is known to measure under conditions of the art, for example at room temperature (e.g.at or about 20 ℃) in a buffer (e.g.a buffer which is stable at pH 7.5, such as a borate (e.g.0.02M) or Tris/hydrochloric acid (e.g.at 0.01M) buffer) in water and in the presence of DNA at a concentration of 10 to 30. mu.M (e.g.20. mu.M). In addition, the dissociation constant can be estimated by comparing the binding affinity of a compound to a set of DNA sequences and the binding affinity of a well-known compound (e.g., distamycin) to the same sequences.

Unless otherwise specified, the term "DNA" refers to double-stranded DNA. Further, as used herein, the term "DNA sequence" includes any oligomer or polymer of DNA that partially (or completely) spans three or more base pairs.

Particular embodiments of the invention are mentioned in relation to compounds of formula I wherein:

when used herein, R¹¹And R¹²Each independently represents H or C_1-6Alkyl (such as R)¹¹And R¹²Both represent H).

Particular embodiments of the present invention relate to compounds of formula I, wherein:

(a) the compounds of formula I contain at least one structural fragment of Q different from formula Ia^aOr Q^bA group;

(b) the compounds of formula I contain a compound representing naphthyl (optionally selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4Substituted with one or more substituents of alkoxy), Het³Or at least one Q of a structural fragment of the formula Id or, in particular, of a structural fragment of the formula If^aOr Q^bAnd (4) a base.

Other particular embodiments of the present invention relate to compounds of formula I, wherein:

(i)Q^arepresents a structural fragment of the general formula Ia or If (e.g. a structural fragment of the general formula If), Het³Or naphthyl (except speciallyUnsubstituted naphthyl, which may optionally be substituted in the latter radical by radicals selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4Substituted with one or more substituents of alkoxy);

(ii)Q^brepresents Het³Or, in particular, a structural fragment of the formula Ia, Id or If.

Other particular embodiments of the present invention relate to compounds of formula I, wherein:

(I) when Q is^aRepresents a structural fragment of the formula Ia, Q^bRepresents Het³Or, in particular, a structural fragment of the formula Id or If (e.g.a structural fragment of the formula If);

(II)Q^aand Q^bOne of them represents a structural fragment of the general formula If, and Q^aAnd Q^bAnd the other represents naphthyl (optionally substituted by one or more substituents selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4Substituted with one or more substituents of alkoxy), Het³Or, in particular, structural fragments of the formulae Ia, Ib, Ic, Id or If,

or Q^aRepresents Het³And Q^bRepresents naphthyl (optionally selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4Substituted with one or more substituents of alkoxy), Het³Or a structural fragment of formula Ia, Ib, Ic, Id, Ie or If;

(III)Q^brepresents Het³A structural fragment of formula Id or in particular If.

Still further, particular embodiments of the invention relate to compounds of formula I, wherein:

R¹represents NO₂、-N(R^2a)R^2bOr, in particular, H;

R^1arepresents H or C_1-8An alkyl group;

a represents 1, 2 or 3 (e.g. 1 or 2);

a represents C_2-6Alkylene (e.g. C)_2-3N-alkylene);

d represents Het¹or-N (R)^3a)R^3b；

Het¹Represents a five-to seven-membered heterocyclic group containing at least one N atom and optionally one or more heteroatoms selected from N, O or S, consisting of C_1-4Alkyl or C_1-4Alkoxy (such as Het)¹Represents a six-membered heterocyclic ring containing one N atom and an O or S atom, thereby forming, for example, a thiomorpholine, or, in particular, a morpholine ring) is optionally substituted;

R^3aand R^3bRepresents C_1-4Alkyl (e.g., methyl);

Q^aand Q^bIndependently represent

Naphthyl (optionally selected from halo, nitro, N (R)^2a)R^2b、C_1-3Alkyl or C_1-3One or more substituents of alkoxy groups being optionally substituted) (e.g., unsubstituted naphthyl, such as unsubstituted naphthalen-2-yl),

Het³，

or a structural fragment of the formula Ia, Id or If,

Q^crepresents a structural fragment of formula Ia, Ib, Ic, Id or If (e.g. a structural fragment of formula Ia or Id);

R⁴represents H;

R⁵represents C_1-6Alkyl (e.g., methyl);

R⁸represents H or, in particular, C_1-8Alkyl (e.g. C)_3-7Alkyl, such as isopropyl or, in particular, 3-methylbutan-1-yl);

R¹⁰each independently represents OH, halo, nitro, N (R)^2a)R^2b、C_1-3Alkyl or C_1-3Alkoxy (e.g. nitro or C)_1-2Alkoxy, such as methoxy);

b represents 0, 1 or 2 (e.g. 0 or 1, when G²When representing N, is 0 or G²Represents CH or C-R¹⁰Is 1);

G¹represents CH;

G²represents CH or N;

Het³represents a ten-membered bicyclic heterocyclic group containing a nitrogen atom and optionally one or more heteroatoms selected from N, O or S, consisting of a group selected from halo (e.g. chloro), nitro, N (R)^2a)R^2b、Het^a、C_1-3Alkyl OR OR^a(e.g., a ten-membered bicyclic heterocyclic group containing one or two N atoms, consisting of a group selected from halo (e.g., chloro), nitro, C_1-2Alkyl OR OR^aOne to three substituents of (a) optionally substituted) is optionally substituted;

R^arepresents C_1-2Alkyl or, especially Het^b；

Het^aAnd Het^bIndependently represent a ten-membered or, in particular, a nine-membered heterocyclic group containing one or more (e.g. one to three) heteroatoms selected from N, O or S, consisting of_1-2Alkyl or C_1-2An oxyalkyl group (e.g. a nine membered bicyclic heterocyclic group containing one, two and especially three N atoms, which group is optionally substituted by one to three substituents selected from halo, methyl or methoxy).

R¹¹And R¹²All represent H;

when used herein, R^2aAnd R^2bEach independently represents H or C_1-2An alkyl group.

Particular embodiments of the present invention relate to compounds of formula I, wherein:

(a)Q^arepresents Het³(e.g., quinolinyl, such as quinolin-2-yl or quinolin-3-yl);

(b)Q^arepresents naphthyl (selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy groups are optionally substituted) (e.g., unsubstituted naphthyl, such as unsubstituted naphthalen-2-yl);

(c)Q^arepresents a structural fragment of formula If;

(d)Q^arepresents a structural fragment of the general formula If, in which G²Represents CH (e.g. phenyl optionally substituted by R in the 3 or 4 position¹⁰In which R is¹⁰As defined hereinbefore (e.g. nitro or especially methoxy);

(e)Q^arepresents a structural fragment of the general formula If, in which G²Represents N (e.g. 3-or 4-pyridine);

(f)Q^arepresents a structural fragment of the formula Ia, in which G¹Represents CH, R⁴And R⁵As defined hereinbefore (e.g. R)⁴Represents H, R⁵Represents a methyl group).

Other particular embodiments of the present invention relate to compounds of formula I, wherein:

(i)Q^brepresents a structural fragment of formula If (e.g., wherein G²A structural fragment of formula If representing CH or N, b representing 0, such as pyridyl (e.g. 2, 5-pyridinylene, e.g. wherein the 2-position of the pyridine ring is bound to a substituent R¹²On the C atom) or phenylene (e.g., 1, 4-phenylene));

(ii)Q^brepresents (e.g. when Q is^aRepresents Het³Or of the formula If) a structural fragment of the formula Ia (for example G)¹Represents CH, R⁴And R⁵As defined hereinbefore (e.g. R)⁴Represents H and R⁵A structural fragment of the general formula Ia representing methyl);

(iii)Q^brepresents (e.g. when Q is^aRepresents a structural fragment of formula If, in particular Het³) Structural fragments of the formula Id (e.g., wherein R⁸Represents C_1-8Alkyl or, in particular, H, a structural fragment of the formula Id);

(iv)Q^bis (such as Q)^aRepresents a structural fragment of the general formula If) Het³(e.g., quinonylene, such as 2, 6-quinonylene, e.g., wherein the 2-position of the quinoline ring is bonded to a substituted group R¹²On the C atom) of (C).

A further particular embodiment of the present invention relates to compounds of the general formula I, wherein:

Q^aand Q^bAre connected in a trans-form with each other,

R¹¹and R¹²Are all represented by H, and are,

a represents 2;

each Q^cIndependently represents a structural fragment of formula Ia or Id as defined above.

Thus, particular embodiments of the invention are mentioned including those wherein the compound of formula I is a compound of formula Ig;

wherein

The wavy line represents optional cis or trans stereochemistry;

R¹represents NO₂、-N(R^2a)R^2bOr, in particular, H;

a represents 1 or, in particular, 2;

Q^arepresents naphthyl (selected from OH, halo, cyano, nitro, N (R)^2a)R^2b、C_1-4Alkyl or C_1-4One or more substituents of alkoxy being optionally substituted) or especially Het³(e.g., a hydroquinone group such as a benzenediol-2-yl or benzenediol-3-yl group) or a structural fragment of formula If;

G²represents CH or N; and is

R^2a、R^2b、Het³、Q^cA and D are as defined above.

Further particular embodiments of the invention which may be mentioned include those in which, in the compounds of the general formula Ig:

R¹represents H;

a represents 2;

Q^aand contain G²The six-membered rings of (a) are trans-linked to each other;

Q^arepresents

Het³(e.g. hydroquinone radicals, such as hydroquinone-2-yl or hydroquinone-3-yl) or

Structural fragment of formula If (e.g. phenyl from R)¹⁰Optionally substituted in the 3-or 4-position, wherein R¹⁰As defined above (e.g. nitro or, especially, methoxy));

G²represents CH (e.g. when Q^aRepresents Het³When Q is not satisfied) or N (e.g. when Q is satisfied)^aRepresents a structural fragment of formula If);

each Q^cRepresents a structural fragment of formula Ia;

G¹represents CH;

R⁴represents H;

R⁵represents C_1-6Alkyl (e.g., methyl);

a represents C₃N-alkylene or, especially, C₂An n-alkylene group;

d represents-N (R)^3a)R^3bOr, especially Het¹；

Het¹Represents a six-membered heterocyclic ring containing one N atom and an O or S atom, thus forming, for example, a thiomorpholine or, in particular, a morpholine ring;

R^3aand R^3bRepresents C_1-4Alkyl (e.g., methyl).

Specific embodiments of the invention are mentioned including the specific example compounds disclosed below.

A further embodiment of the invention relates to compounds of the general formula I as defined above, but wherein:

a represents 0, 1, 2, 3 or 4; or

a represents 0.

Preferred compounds of formula I include those that bind to the minor groove of DNA.

The affinity of the DNA can be measured by techniques well known to those skilled in the art, such as capillary electrophoresis. Furthermore, the affinity of a piece of DNA can be measured by techniques well known to those skilled in the art, such as DNA footprinting.

Preparation of

The compounds of the invention may be prepared by techniques well known to those skilled in the art (e.g., polypeptide synthesis), using commercially available starting materials well known in the literature, or starting materials obtained by analogous processing methods as described below, or by conventional synthetic procedures that conform to standard techniques and use appropriate reagents and reaction conditions.

According to the present invention there is also provided a process for the preparation of a compound of formula I, which process comprises:

(a) reacting a compound of formula III with a compound of formula IV:

wherein Q^cD and A are as defined above and, c is as defined below,

wherein L is¹Represents a leaving group (e.g. OH, halo (e.g. Cl or Br) or-OC (O) R¹⁴Wherein R is¹⁴Represents C_1-6Alkyl radical, Q_1-6Alkenyl radical, C_5-6Cycloalkyl or aryl (which in the latter group is selected from halo, cyano, nitro, C_1-4Alkyl or C_1-4One or more substituents of alkoxy are optionally substituted), c and d are each integers from 0 to 4, wherein the sum of c and d is from 1 to 4 and is, for example, under conditions well known to those skilled in the art (e.g.: (i) when L is¹Represents OH in the presence of a coupling agent (e.g. oxalyl chloride in DMF, EDC, DCC, HBTU, HATU, PyBOP or TBTU), a suitable base (e.g. pyrimidine, DMAP, TEA, 2, 4, 6-trimethylpyridine or DIPEA) and a suitable organic solvent (e.g. DCM, MeCN, EtOAc or DMF); or (ii) when L¹Represents halogen or OC (O) R¹⁴In the presence of a suitable base (e.g. pyrimidine, DMAP, TEA, 2, 4, 6-trimethylpyrimidine or DIPEA) and a suitable organic solvent (e.g. DCM, MeCN, EtOAc or DMF) R¹、R¹¹、R¹²、Q^aTo Q^cAnd aryl is as defined above;

(b) reaction of a Compound of formula Va with a Compound of formula VI

Wherein R is¹⁵Represents aryl (the latter group of which is selected from halo, cyano, nitro, C_1-4Alkyl or C_1-4Alkoxy) or C_1-6One or more substituents of alkyl are optionally substituted), and R¹、R¹¹、Q^aAnd the aryl group is as defined above,

wherein A, a, D, R¹²Q^bAnd Q^cAs defined above, for example under conditions well known to those skilled in the art (e.g. in the presence of a suitable organic solvent (e.g. diethyl ether, THF, toluene));

(c) reacting a compound of formula Vb with a suitable base such as sodium hydride,

wherein R is¹⁶Represents C_1-6Alkyl, and R¹、R¹¹、Q^aAnd aryl are as defined above, e.g. under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene)), followed by reaction with a compound of formula VI as defined above, e.g. under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene));

(d) reacting a compound of formula VIIa with a compound of formula VIII,

wherein R is¹²、R¹⁵、Q^b、Q^cA, A and D are as defined above,

R¹-Q^a-C(O)R¹¹ VIII

wherein R is¹、R¹¹And Q^aAs defined above; for example under conditions well known to those skilled in the art (as described in method (b) above), or

(e) The compound of formula VIIb is reacted with a suitable base,

wherein R is¹²、R¹⁶、Q^b、Q^cA, A and D are as defined above, for example under conditions well known to those skilled in the art (as described in method (c) above), followed by reaction with a compound of formula VIII as defined above, for example under conditions well known to those skilled in the art (as described in method (c) above).

In another embodiment of the process for preparing the compound of formula I according to process (a) above, the integer c for the compound of formula III above and the integer d for the compound of formula IV above are both integers from 0 to 4, wherein the sum of c and d is from 0 to 4.

The compounds of formula IV can be prepared as follows:

(i) a compound of formula Va as defined above with a compound of formula IX:

or a protected derivative thereof, wherein L¹、Q^b、Q^cD and R¹²As defined above, for example under conditions well known to those skilled in the art (as described in method (b) above), followed (if desired) by the addition of an L¹Conversion of a group to another (e.g., OH to halo, etc.);

(ii) reaction of a compound of formula Vb as defined above with a suitable base (e.g. sodium hydride), for example under conditions well known to those skilled in the art (as described in method (c) above), followed by reaction with a compound of formula IX as defined above (for example under conditions well known to those skilled in the art as described in method (c) above);

(iii) a compound of the general formula Xa,

or a protected derivative thereof, with a compound of the general formula VIII as defined above, in which L¹、R¹²、R¹⁵、Q^b、Q^cAnd d is as defined above, e.g. under conditions well known to the person skilled in the art (as described in method (b) above); or

(iv) A compound of the general formula Xb,

or a protected derivative thereof, with a suitable base (e.g. sodium hydride), wherein L¹、R¹²、R¹⁶、Q^b、Q^cAnd d is as defined above, for example under conditions well known to those skilled in the art (such as when in the presence of a suitable solvent (as described in method (c) above)), followed by reaction with a compound of formula VIII as defined above, for example under conditions well known to those skilled in the art (such as described in method (c) above).

Wherein Q^bRepresents quinoline (Het)³) Or a pyrimidine (structural fragment of the general formula If) linked to C (R) via the 2-position of the ring system¹²) And R is¹¹And R¹²Compounds of the formula IV, which both represent H, can be prepared by reacting a compound of the formula XI,

or a protected derivative thereof, wherein Q^xRepresents an optional fused benzene ring and L¹、Q^cD and aryl are as defined above, with a compound of formula XII,

wherein R is¹And Q^aAs defined above, for example under conditions well known to those skilled in the art (e.g. as described in US 4,009,174, such as in the presence of acetic anhydride and a suitable catalyst (e.g. zinc chloride), and optionally in the presence of a suitable solvent (e.g. an aromatic hydrocarbon such as xylene)).

Compounds of formula Va can be prepared by compounds of formula XIII,

wherein L is³Are halo (e.g. Br, Cl) and Q^a、R¹And R¹¹As defined above, with a compound of formula XIV,

P(R¹⁵)₃ XIV

wherein R is¹⁵As defined aboveAs defined herein, the compound of formula Va obtained is used without isolation (e.g. in a "one-shot" preparation of a compound of formula I selected from compounds of formula XIII), or alternatively, isolated prior to use, e.g. under conditions well known to the person skilled in the art, such as in the presence of a suitable solvent (e.g. diethyl ester, THF, toluene) and a suitable base (e.g. sodium methoxide, n-butyllithium, sodium hydride).

The compound of formula Vb may be prepared by reaction of a compound of formula XIII as defined above with a compound of formula XV,

P(O)(OR¹⁶)₃ XV

wherein R is¹⁶As defined above, for example under conditions well known to those skilled in the art (such as in the presence of a suitable solvent (e.g., diethyl ether, THF, toluene)).

The compounds of formula VI can be prepared by reacting a compound of formula XVI,

wherein Q^b、Q^cA, A and D are as defined above, with a compound of general formula XVII,

wherein L is³And R¹²As defined above, e.g. under conditions well known to the person skilled in the art (e.g. in the presence of a suitable catalyst (e.g. FeCl)₃Or AlCl₃) And in the presence of a suitable solvent (e.g., nitrobenzene, methylene chloride).

Compounds of the formula VIIa can be prepared by compounds of the formula XVIII

Wherein L is³、Q^b、Q^cA, D, a and R¹²The reaction of a compound of the formula XIV as defined above, with a compound of the formula XIV as defined above, is carried out, for example, under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene) and a suitable base (e.g. sodium methoxide, n-butyllithium, sodium hydride)) wherein the compound of the formula VIla obtained can be used without isolation (e.g. in a "one-shot" preparation of a compound of the formula I selected from compounds of the formula XVIII) or, alternatively, isolated prior to use.

The compound of formula VIIb may be prepared by reaction of a compound of formula XVIII, as defined above, with a compound of XV, as defined above, for example under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent such as diethyl ether, THF and toluene).

The compounds of formula IX can be prepared by reacting a compound of formula XIX,

or a protected derivative thereof, wherein Q^b、Q^cD and L¹As defined above, and a compound of formula XVII as defined above, for example under conditions well known to those skilled in the art (such as those described above in connection with the synthesis of compounds of formula VI).

The compounds of formula Xa may be prepared by reacting a compound of formula XX,

or a protected derivative thereof, wherein Q^b、Q^c、L¹、L³D and R¹²The reaction of a compound of formula XIV as defined above, with a compound of formula XIV as defined above, is carried out, for example, under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene) and a suitable base (e.g. sodium methoxide, n-butyllithium, sodium hydride), wherein the obtained compound of formula Xa is used without isolation (e.g. in a "one-shot" preparation of a compound of formula IV from a compound of formula XX) or, alternatively, is isolated prior to use).

Compounds of formula Xb may be prepared by reaction of a compound of formula XX as defined above and a compound of formula XV as defined above, for example under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene)).

Compounds of formula XIII can be prepared by reacting compounds of formula XXI,

wherein R is¹、Q^aAnd R¹¹As defined above, and a reducing agent (e.g., LiAlH)₄、NaBH₄) For example in the presence of a suitable solvent (for example diethyl ether, THF, toluene), followed by the resulting intermediate alcohol and the appropriate halogen atom (for example in combination with a suitable catalyst (for example ZrCl) under conditions well known to the person skilled in the art (for example in the presence of a suitable solvent (for example diethyl ether, THF, toluene))₄) Thionyl chloride or sodium iodide) to-OH, for example under conditions well known to those skilled in the art, such as in the presence of a suitable solvent, such as diethyl ether, THF or acetonitrile.

The compound of formula XVIII may be prepared by reacting a compound of formula VI as defined above with a reducing agent (e.g. LiAlH)₄、NaBH₄) ToShould, for example, be prepared by subsequent reaction of the resulting intermediate alcohol and a reagent suitable for effecting substitution of the halogen atom (as described above) for the-OH, under conditions well known to those skilled in the art, such as in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene).

The compounds of the formula XX can be prepared by reaction of the corresponding compounds of the formula IX as defined above with a reducing agent (e.g.LiAlH)₄、NaBH₄) For example under conditions well known to those skilled in the art (e.g. in the presence of a suitable solvent (e.g. diethyl ether, THF, toluene)), followed by reaction of the resulting intermediate alcohol with a reagent suitable for effecting substitution of the halogen atom (as described above) for-OH.

Compounds of formula XXI can be prepared by compounds of formula XXII,

R¹-Q^a-H XXII

wherein R is¹And Q^aAs defined above, with a compound of formula XXIII,

wherein L is³And R¹¹As defined above, e.g. under conditions well known to the person skilled in the art (e.g. in the presence of a suitable catalyst (e.g. FeCl)₃Or AlCl₃) And in the presence of a suitable solvent (e.g., nitrobenzene, methylene chloride).

Compounds of formulae III, VIII, XI, XII, XIV to XVII, XIX, XXII, XXIII and derivatives thereof are either commercially available as known in the literature or obtained in analogous methods as described herein or from WO 2003/059881, or obtained by conventional synthetic methods according to standard techniques from readily available starting materials using appropriate reagents and reaction conditions.

Substituents on aryl (e.g., phenyl) and heterocyclic groups of the compounds defined herein may be converted to other desired substituents using methods well known to those skilled in the art. For example, a hydroxy group may be converted to an alkoxy group, a phenyl group may be halogenated to provide a halophenyl group, a nitrogen may be reduced to form an amino group, a halo group may be replaced by a cyano group, and the like.

Those skilled in the art will also appreciate that different standard substituents or functional groups that are interconverted and transformed within certain compounds of formula I will yield other compounds of formula I. For example, chlorine may be replaced by alkoxy, aryloxy or heteroaryloxy, the carbonyl group may be reduced to a hydroxyl or methylene group and the hydroxyl group may be converted to a halo group.

The compounds of the present invention may be isolated from their reaction mixtures by conventional techniques.

In the above process, it is possible or possible for the person skilled in the art that the functional groups of the intermediate compounds may or may need to be protected by protecting groups.

Functional groups to be protected include hydroxyl, amino and carboxylic acid. Suitable protecting groups for the hydroxyl group include trialkylsilyl and diarylalkylsilyl groups (e.g.tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl and alkylcarbonyl groups (e.g.methyl and ethylcarbonyl). Suitable protecting groups for the amino group include benzyl, t-butoxycarbonyl, 9-fluoromethoxycarbonyl or benzyloxycarbonyl. Suitable protecting groups for carboxylic acids include C_1-6Alkyl or benzyl esters.

The protection or deprotection of the functional groups may be carried out before or after any of the above described reaction steps.

The protecting group may also be removed according to methods well known to those skilled in the art and techniques described below.

The use of protecting Groups is described in detail in J.W.F.McOmie, Plenum Press (1973) editions "Protective Groups in Organic Chemistry" and "Protective Groups in Organic Synthesis", 3rd edition, T.W.Greene & P.G.M.Wutz, Wiley-Interscience (1999).

It will be appreciated by those skilled in the art that in order to obtain the compounds of the invention by other routes and in an in some cases more convenient methods, each of the steps mentioned herein may be performed in a different order and/or each reaction may be performed at a different stage in the overall route (i.e., substituents may be added to the different intermediates described above in connection with the particular reaction and/or chemical transformations may be performed thereon). This will depend in particular on factors such as the nature of the other functional groups present in the particular substrate, the main intermediates and the availability of strategies (if any) to employ protecting groups. Clearly, the type of chemistry involved will influence the choice of reagents used in the above synthetic steps, the need and type of protecting groups employed, and the order in which the synthesis is performed.

Use and pharmaceutical preparation

The compounds of the present invention are valuable because they possess pharmaceutical activity. They are therefore useful as pharmaceuticals.

Thus, according to a further aspect of the invention, the compounds of the invention may be used as pharmaceuticals.

In particular, the compounds of the invention bind to DNA and thus may replace or inhibit the binding of enzymes or other regulatory proteins to DNA. Enzymes mentioned in this connection include enzymes necessary for replication (thus providing an effect of inhibiting DNA replication), enzymes involved in transcription (thus providing an effect of inhibiting the expression of certain peptides (proteins, enzymes, etc.)).

Thus, in a further aspect, the invention provides a method of inhibiting DNA replication, which method comprises contacting DNA with an inhibitory amount of a compound of the invention.

Due to the ability of the compounds to inhibit DNA replication (e.g., inhibit transcription by blocking binding or replacement of regulatory proteins or DNA-enzyme complexes, such as reverse transcriptase or topoisomerase), the compounds of the invention can be used to treat diseases that rely on DNA replication for transmission. Such diseases include cancer and diseases involving viruses, bacteria, fungi or other microorganisms (e.g., parasitic diseases such as malaria).

Thus viewed from a further aspect the invention provides a method of treating a disease which is dependent on DNA replication for transmission, such as cancer or a viral, bacterial, fungal or other microbial infection, which method comprises administering a compound of the invention in a therapeutically effective amount to a patient suffering from the disease. Such treatment would be particularly useful for patients suffering from an immune deficiency.

Because they have different modes of action against many conventional antiviral, antibacterial, antifungal or other antimicrobial (e.g., antiparasitic) agents, the compounds of the present invention are particularly useful in the treatment of viral, bacterial, fungal or other microbial (e.g., parasitic) infections when the infectious agent is resistant to one or more antiviral, antibacterial, antifungal or antimicrobial (e.g., antiparasitic) drugs that have different modes of action. From this aspect, according to a further aspect, the present invention provides a method of treating a viral, bacterial, fungal or other microbial (e.g. parasitic) infection, wherein the source of the infection by the virus, bacteria, fungus or other microbial (e.g. parasitic) is resistant to one or more antiviral, antibacterial, antifungal or antimicrobial (e.g. antiparasitic) drugs, respectively, that do not act by inhibiting DNA replication, the method comprising administering to a human suffering such infection a therapeutically effective amount of a compound of the present invention.

In addition to being useful by themselves in the treatment of diseases that rely on DNA replication for transmission, the compounds of the present invention may be used in combination with one or more other compounds or treatment regimens useful in the treatment of such diseases. Thus, according to a further aspect of the invention, there is provided a method of treating a disease which is mediated by DNA replication (such as cancer or a viral, bacterial, fungal or other microbial infection) which comprises administering to a patient a therapeutically effective amount of a compound of the invention in combination with one or more other therapeutic agents known to be effective against the disease.

As used herein, the term "combination" encompasses the use of other known agents that are effective in treating the disease prior to, during and/or after the use of the compounds of the present invention. When more than one agent is used, the term also encompasses the use of different other drugs at different times than the use of the compounds of the present invention.

Known agents for treating diseases that rely on DNA replication for transmission (e.g., anti-tumor, anti-viral, anti-bacterial, anti-fungal, or anti-other microbial (e.g., anti-parasitic) include those listed under The relevant headings "Martindale: The Complete Drug Reference," 32 th edition, The Pharmaceutical Press, London (1999)), which are incorporated herein by Reference.

Anti-cancer drugs also include non-chemical agents such as ionizing radiation (e.g., subatomic particle radiation such as alpha particles, beta particles, neutrons, protons, mesons, and heavy ions or electromagnetic radiation such as high frequency X-rays or gamma rays). Chemical anti-cancer agents that are mentioned include:

(a) an alkylating agent comprising:

(i) nitrogen mustards, e.g. mechlorethamine (HN)₂) Cyclophosphamide, ifosfamide, melphalan (L-propylbenzene mustard), and chlorambucil;

(ii) aziridines and analgin, such as hexamethylmelamine, thiotepa;

(iii) alkyl sulfonates and thiosulfates such as busulfan, Methyl Methanesulfonate (MMS) and methyl methane thiosulfate (methyl methane thiosulfonate);

(iv) nitrosoureas and nitrosoguanidines, such as carmustine (BCNU), lomustine (CCNU), nitrosomethylurea (methyl-CCNU), streptozotocin (streptozotocin), and N-methyl-N' -nitro-N-nitrosoguanidine (MNNG); and

(v) triazenes, such as dacarbazine (DTIC, dimethyltriazinylimidazole-amide).

(b) An antimetabolite comprising:

(i) folic acid analogs, such as methotrexate (amethopterin);

(ii) pyrimidine analogs such as fluorouracil (5-fluorouracil; 5-FU), fluorouracil deoxynucleoside (FUdR), and cytarabine (cytosine arabinoside); and

(iii) purine analogs and related inhibitors, such as mercaptopurine (6-mercaptoprine; 6-MP), thioguanine (6-thioguanine; TG) and pentostatin (2' -deoxykomycin).

(c) A natural product comprising:

(i) vinblastine such as Vinblastine (VLB) and vincristine;

(ii) etoposide, such as etoposide and teniposide;

(iii) antibiotics, including dactinomycin (actinomycin A, C, D or F), daunorubicin (daunomycin; daunomycin), doxorubicin, bleomycin, mithramycin (milamycin), and mitomycin (mitomycin A, B or C);

(iv) enzymes such as levo-asparaginase; and

(v) biological response modifiers such as interferon alphaphenomers.

(d) Various reagents, including:

(i) platinum coordination complexes, such as cisplatin (cis-DDP) and carboplatin;

(ii) anthracenediones, such as mitoxantrone and anthracyclines;

(iii) substituted ureas, such as hydroxyurea;

(iv) methylhydrazine derivatives such as procarbazine (N-procarbazine, MIH);

(v) adrenocortical suppressants, such as mitotane (o, p' -DDD) and amiperidone;

(vi) sequoyitol and analogs/derivatives;

(vii) hormone agonists and antagonists, including flutamide and tamoxifen;

(viii) photosensitizing compounds (e.g., psoralen);

(ix) DNA topoisomerase inhibitors (e.g., methacridine and camptothecin);

(x) Anti-angiogenic drugs (e.g., SU6668, SU5416, combretastatin a4, angiostatin, and endostatin); and

(xi) Immunotherapeutics (e.g. radiolabeled antibodies such as Bexxar^TMAnd Theragyn^TM(Pemtumomab^TM)).

The antiviral drugs mentioned include acyclovir, gancyclovir, AZT, ddI, amantadine hydrochloride, isopropyl creatinine, vidarabine and the like.

Antibacterial agents which may be mentioned include natural and synthetic penicillins and cephalosporins, sulfonamides, erythromycin, kanamycin, tetracycline, chloramphenicol, rifampin and antibiotics gentamicin, ampicillin, benzylpenicillin, benzathine, benzyloxyethylpenicillin, benzyloxymethylpenicillin, procainazine, cloxacillin, flucloxacillin, methicillin sodium, amoxicillin, bacampicillin hydrochloride, cyclohexacillin, mezlocillin, pimaricillin, ampicillin peptidolipidium hydrochloride, carbenicillin sodium, piperacillin, ticarcillin, mecillin, pirecellinan, cefaclor, cephalexin, cefotaxime, cefoxitin, cefixime, cefuroxime, cephalexin, cephalotin, cefamandole sodium, cefazolin, cephamotrimadin, doxycycline, aztreonam, chlortetracycline hydrochloride, sodium oxymetamycin, demeclocycline hydrochloride, doxycycline, lymecycline, minocycline, oxytetracycline, amikacin, neomycin B, neomycin, netilmicin, tobramycin, polymyxin E, sodium fusidate, polymyxin B, spectinomycin, vancomycin, calcium sulphanilate, sulfalene, sodium sulfadiazine, sulfadimidine, sulfaguanidine, sulfonylurea, capreomycin, metronidazole, tinidazole, cinoxacin, ciprofloxacin, nitrofurantoin, urotropin, streptomycin, hydroxybenzyl, polymyxin M, polymyxin B, furazolidone, nalidixic acid, trimethoprim-sulfamethoxazole, clindamycin, lincomycin, cycloserine, isoniazid, butanol, ethionamide, pyrazinamide, and the like.

The antifungal agents to be mentioned include miconazole, ketoconazole, itraconazole, fluconazole, fusidic acid, amphotericin, flucytosine, griseofulvin, natamycin, nystatin and the like.

Antiparasitic drugs (antimalarial drugs) mentioned include pyrimethamine, proguanil, chloroquine, primaquine, mefloquine, quinine, tetracycline, atovaquone, artemisinin, dihydroartemisinin, artemether, artesunate and salts thereof and sulfonamides.

When the compounds of the present invention are administered to a patient in combination with one or more agents known to be effective in treating diseases that rely on DNA replication, the compounds of the present invention and the other agents may conveniently be administered separately as a single component. Thus according to a further aspect of the invention there may be provided a combination product comprising the following components:

(A) formulations comprising the compounds of the invention; and

(B) including one or more other chemical formulations known to be effective in treating diseases that rely on DNA replication.

According to this aspect of the invention, the combination product provides for the use of a compound of the invention in combination with one or more other agents effective in the treatment of diseases that are dependent on DNA replication and therefore may be presented (i.e. formulated) as separate components (i.e. (A) and (B) are used alone) or as a combined preparation (i.e. a single formulation of a compound of the invention and one or more other agents known to be effective in the treatment of diseases that are dependent on DNA replication). When the components (a) and (B) are used as mutually independent components, the combination product is alternatively referred to as a "combination kit".

Other drugs known to be effective in treating diseases that rely on DNA replication to transmit disease in this aspect of the invention include the drugs mentioned above. Thus, in a preferred embodiment of the invention, other chemical agents effective in treating diseases that rely on DNA replication to transmit disease include one or more chemical anti-neoplastic, anti-viral, anti-bacterial, anti-fungal and/or anti-parasitic agents (such as those mentioned above).

In a further preferred embodiment of the invention, each of components (a) and (B) is prepared in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

The compounds of the invention can be conveniently administered in the form of pharmaceutical preparations containing the active ingredient in free radical or non-toxic organic or inorganic acid addition salts, in pharmaceutically acceptable dosage forms, by oral, subcutaneous, intravenous, arterial, transdermal, intranasal, inhalation or any other parenteral route. The components may be administered in different doses depending on the condition, the patient to be treated and the route of administration.

According to a further aspect of the invention there may be provided a pharmaceutical formulation comprising a compound of the invention in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier. Such formulations may be used to treat diseases that rely on DNA replication for transmission. Accordingly, in one embodiment the present invention provides a pharmaceutical formulation comprising a compound of the invention in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier, and one or more other chemical agents known to be effective in the treatment of diseases which rely on DNA replication for transmission (e.g. one or more chemical anti-tumour, anti-viral, anti-bacterial, anti-fungal and/or anti-parasitic agents such as those mentioned above).

Suitable daily dosages of the compounds of the present invention for the treatment of humansIs about 1 to 2000mg/m²。

The most effective mode of administration and dosage of the compounds of the invention will depend upon several factors including the particular condition being treated, the extent and location of the condition in the patient being treated, and the health of the patient being treated and the response to the administration of the compound. Thus, the dosage of the compounds of the present invention administered will vary from one person to another. Methods for determining an appropriate dosage for an individual are well known to those skilled in the art.

In addition to being applicable to the treatment of disease, the compounds of the invention are also useful in a variety of assays that rely on DNA binding. For example, it is now known that compounds that bind to the minor groove of DNA can stabilize DNA duplexes, which can more stably match DNA duplexes than mismatched DNA duplexes, thus allowing for convenient discrimination between matched and mismatched DNA duplexes (e.g., in terms of duplex melting temperatures).

Thus, according to a further aspect of the invention, there is provided a method of stabilising a DNA duplex formed between a first and a second DNA single strand by contacting the compound of the invention with the DNA duplex.

In addition, the present invention provides a method of increasing the difference between the melting temperatures of a first strand and a second strand of DNA, wherein each DNA duplex consists of the same first single strand in each DNA duplex and a different second single strand in each DNA duplex, comprising contacting each DNA duplex with a compound of the present invention. In a preferred embodiment, the first DNA duplex has a higher degree of base pair match (e.g., a perfect match) than the second DNA duplex containing at least one base pair mismatch.

Compounds that stabilize perfectly matched DNA duplexes to a higher degree than mismatched DNA duplexes may be used to reduce the level of "false positives" in DNA hybridization analysis techniques, such as the technique described in US 6,221,589, which is incorporated herein by reference. Reduced "false positive" results can be obtained by using more stringent conditions followed by hybridization conditions (e.g., high rinse temperatures) in the presence, but not in the absence, of a compound that stabilizes the duplex. Accordingly, the present invention provides a method of increasing the maximum temperature of a wash followed by a DNA hybridization reaction, which comprises supplying a compound of the present invention to the hybridization reaction mixture. As used herein, the term "maximum temperature of washing subsequent to DNA hybridization reaction" refers to the highest possible washing temperature that does not result in substantial loss of a "true positive" result (i.e., a perfect or highest match DNA duplex).

When the above methods involving DNA duplexes are used herein, the term "contacting" includes mixing a compound of the invention with a DNA duplex. However, the term also includes attaching (e.g., covalently bonding) a compound of the invention (e.g., a compound of the invention with a haloalkyl group), or a derivative thereof (e.g., a compound of formula V) with a functional group (e.g., a carbonyl, amino, or carboxylic acid group) that can be used to form a suitable linkage, to one or both of the single-stranded DNAs that form the duplex. The "labeled" single stranded DNA may be used as primers, capture probes, or in a variety of different assays (e.g., capture detection assays, 5' nuclease assays, and indicator assays).

The compounds of the invention also have fluorescent properties. The fluorescent compounds of the present invention may function in a variety of assays that rely on DNA binding involving or requiring fluorescence.

Thus, according to a further aspect of the invention, there is provided a method of detecting dsDNA in a sample, the method comprising contacting a compound of the invention with the sample and comparing the fluorescence of said compound contacted with said sample with the fluorescence of said compound independently, a change in fluorescence being indicative of the presence of DNA in the sample.

In particular embodiments of the compounds of the invention, the change in fluorescence can be, for example, a change in the wavelength of light emitted by the compounds of the invention, a change in the wavelength of light absorbed by the compounds, or a change in the intensity of light emitted by the compounds. Additionally, in certain embodiments, the dsDNA may be fluorescently labeled. When labeled by this method (and even when not so labeled), the dsDNA can serve as a donor or acceptor in a "FRET" type assay that detects the presence of dsDNA.

In another embodiment of the invention, a method is provided for detecting and viewing dsDNA in a dsDNA-containing sample, the method comprising contacting the sample with a compound of the invention and illuminating the sample with ultraviolet light and then viewing the dsDNA. In a particular embodiment of the invention, the sample may be selected from an agarose gel electrophoresis experiment or a DNA microarray.

The compounds of the invention may have the particular advantage that once bound to the dsDNA minor groove, their dissociation is relatively small (e.g. dissociation coefficient for dissociation from the DNA minor groove is in the range of 0.1 to 10 nM), indicating that the pharmacokinetic effect of the compounds may be maintained significantly longer than suggested by plasma concentrations in vivo. For a discussion of this effect, see Nucleic Acid Res.26, 3053-3058 (1998); and Chapter 2 of pharmaceutical and Metabolism in drug Design, Smith et al, Mannhold et al, eds, Wiley-VCH, Weinheim, 2001.

In connection with the above, the dissociation constant from the minor groove of dsDNA can be determined, for example, by determining the dissolution temperature of a mixture of various compounds of the invention and DNA, or by microcalorimetry measurements.

Compared with the known compounds, the compounds of the invention have the following advantages: they are more potent, less toxic, more broadly active, more potent, longer acting, produce fewer side effects, are more easily absorbed, or have other useful pharmacological properties.

Biological assay

The effect of the compounds of the invention on inhibiting the growth of various microorganisms is determined by methods well known to those skilled in the art, for example in vitro as described in J.Med.chem.47, 2133-.

In vitro assay

In particular, The Minimum Inhibitory Concentration (MICs) of a compound of The invention against a microorganism (e.g., staphylococcus aureus (s. aureus), Streptococcus faecalis (Streptococcus faecalis), Aspergillus niger (Aspergillus niger), Candida albicans (Candida albicans), or mycobacterium fortuitum) can be determined using methods such as those described in a.j. dry and r.d. wave "The degradation of microbial methods for phytochemical screening" Recent res.device.phytochem.4, 143-152(2000), The disclosures of which are incorporated herein by reference.

Sample concentration the test sample (2mg) was typically dissolved in sterile water to provide a working concentration of 200. mu.g/mL. Test wells on 96-well microtiter plates were inoculated with Mueller-Hinton medium (100. mu.L) for antibacterial analysis and then with Sabouraud medium for antifungal analysis. Solutions (100 μ L) of each test sample were added to one row of each plate and a series of two-fold dilutions were made for successive rows. Incubation was at 37 ℃ for antibacterial assays and at 25 ℃ for antifungal assays. Plates were visually inspected for growth and 7-hydroxy-3H-phenoxazin-3-one-10-oxo was added to each well; a distinct color change from blue to red indicates the occurrence of growth in each individual well. The MIC was determined from the observed color pattern. All tests included sterility and growth controls.

In vivo testing

Toxicity of the compounds of the invention can be determined by direct and indirect methods well known to those skilled in the art, such as those described in J.Imunol. methods 94, 57-63(1986), the disclosures of which are incorporated herein by reference.

With respect to the method described in J.Med.Microbiol.46, 208-213(1997), compounds can be evaluated in vivo in a model of the S.aureus LS-1 strain, which when injected intravenously into mice consistently caused transient bacteremia by joint immobilization for 3-4 days. Normal control mice show inflammation in 50-60% of the joints within 3-4 days. The severity of joint sepsis was determined by measuring the diameter of the affected joint with a caliper. Five groups of mice, typically weighing an average of between 18-28g (5 mice per group), were used for each test compound, where each group was infected with staphylococcus aureus and then provided a metered amount of test compound (one group served as a control).

Examples

Routine experimental procedures

Electrospray mass spectrometry (ES-MS) inVG platform LC-MS. Electron impact (EI-MS) and fast atom bombardment (FAB-MS) mass spectrometryJMS-AX505HA on a mass spectrometer. The exact mass was recorded on a Jeol JMS-700 Mstation high resolution sector magnetic field mass spectrometer, university of Glasgow.

NMR spectroscopy was performed on¹H of 400MHzObtained on an AMX 400 spectrometer. In that¹In H NMR spectra, the abbreviation ` exch ` means that the relevant resonance is in use D₂O disappears when the solution is treated.

HPLC purification of the final product was achieved using Vydac protein and peptide C18 columns on a gradient elution system. The solvent is A: water + 0.1% trifluoroacetic acid (TFA), and B: acetonitrile + 0.1% TFA. All final products obtained after HPLC purification were lyophilized and obtained as TFA salts.

IR spectrum: by usingImpact 400D, KBr pellet and liquid run solid with membrane method.

For column chromatographySilica gel (200 and 400 meshes).

Preparation 1

4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]-1H-pyrrole-2-amides

(i)1-methyl-4-nitro-1H-pyrrole-2-carboxylic acid

(see, e.g., Suckling, C.J., Khalaf, A.I., Pitt A.R., Scobie, M., Tetrahedron, 2000, 56, 5225.)

HNO₃(70%) (8.4mL) was added dropwise to anhydrous acetic acid (40mL) at-25 deg.C and stirred for an additional 20 minutes. This solution was added dropwise to commercially available 1-methyl-1H-pyrrole-2-carboxylic acid (Aldrich) (7.74g, 61.9mmol) in anhydrous acetic acid (60mL) at-25 ℃ and returned to room temperature over a period of 2 hours. The solution was cooled to-40 ℃ at which point a precipitate formed. Before drying under reduced pressure to give the desired product (2.21g, 21%), it was collected and washed with hexane.

m.p.＝199-201℃，(lit.＝199-199.5℃)。

IR(KBr)：3500-2500υ(O-H)，3141υ(N-Me)，2920υ(C-H)，1702υ(C＝O)，1422，1399υ(N-O)，1269υ(C-O)cm^-1。

¹H NMR(CDCl₃)：4.04(3H，s，CH₃)，7.51(1H，d，Ar-H，J＝1.6Hz)，7.71(1H，d，Ar-H，J＝I.6Hz)。

(ii)1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride

(see, e.g., Suckling, c.j., Khalaf, a.i., Pitt a.r., Scobie, m., Tetrahedron, 2000, 56, 5225.)

1-methyl-4-nitro-1H-pyrrole-2-carboxylic acid (0.510g, 3.02 mmol; see step (i) above) and thionyl chloride(7mL) in a round bottom beaker and let the solution in N₂Reflux for 2 hours. The solution was removed under reduced pressure to yield the product as a white to off-white solid (0.556g, 98%).

m.p.＝91-92℃(lit.＝91-92℃)。

IR(KBr)：3126υ(N-Me)，2974υ(Ar-H)，1744υ(C＝O)，1511，1314υ(N＝O)，592δ(C-Cl)cm^-1。

¹H NMR(DMSO)：3.91(3H，s，CH₃)，7.25(1H，d，Ar-H，J＝I.6Hz)，8.22(1H，d，Ar-H，J＝I.6Hz)。

(iii)1-methyl-N- [2- (4-morpholinyl) ethyl]-4-nitro-1H-pyrrole-2-amides

(see, e.g., Kaizerman, j.a., Gross, m.i., Ge, Y, White, S Hu, W, Duan, J, Baird, e.e., Johnson, k.w., Tanaka, r.d., Moser, h.e., Berli, r.w., j.med.chem., 2003, 46, 3914.)

1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride (0.585g, 4.70 mmol; see step (ii) above) in DCM (10mL) was added dropwise over 15 minutes to 2- (4-morpholino) ethylamine (0.673g, 5.17mmol) and NEt in DCM (10mL)₃(0.735. mu.L, 9.42 mmol). The resulting solution was stirred overnight and the reaction was quenched with 5% NaOH solution (20 mL). The layers were separated and the DCM fractions were collected and dried (MgSO)₄) The solution was filtered and removed under reduced pressure to yield the sub-title compound as a white/light yellow solid (1.166g, 88%).

m.p.＝141-143℃，(lit＝143-145℃)。

v_max KBr/cm^-1：3325υ(N-H)，3118，3023υ(Ar-H)，2967，2865，υ(C-H)，1638υ(C＝O)，1539，1311υ(N＝O)，1146υ(C-O-C)。

δ_H ¹H(DMSO)：2.40(4H，m，2(CH₂))，3.31(4H，q，(CH₂)-N-(CH₂)(J＝6.8Hz))，3.56(4H，t，(CH₂)-O-(CH₂)(J＝4.6Hz))，3.89(3H，s，N-Me)，7.39(1H，d，Ar-H(J＝1.6Hz))，8.10(1H，d，Ar-H(J＝1.6Hz))，8.33(1H，tNH(J＝5.6Hz))。

LREIMS: found 283.08(M + H) calculated value C₁₂H₁₈N₄O₄，282.13。

(iv)1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -N- [2- (4-morpholinyl) ethyl]-1H-pyrrole-2-amides

(see, e.g., Kaizerman, j.a., Gross, m.i., Ge, Y, White, S Hu, W, Duan, J, Baird, e.e., Johnson, k.w., Tanaka, r.d., Moser, h.e., Berli, r.w., j.med.chem., 2003, 46, 3914.)

1-methyl-N- [2- (4-morpholinyl) ethyl]-4-Nitro-1H-pyrrole-2-amide (1.42g, 3.41 mmol; see step (iii) above) dissolved in methanol (10mL) and cooled to 0 ℃ Pd/C (0.055g) was added in small portions and the solution was taken up in H₂Stirred for 3 hours. The solution was then filtered and the solvent removed under reduced pressure. 1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride (0.642g, 3.42mmol) dissolved in DCM (10mL) was added and the mixture was stirred at room temperature for 1H. The solvent was removed under reduced pressure and the crude product was separated on basified silica with 1: 1 methanol ethyl acetate to yield the sub-title product.

Yield 0.923g, 67%, m.p. > 230 ℃.

v_max KBr/cm^-1：3339，3284υ(N-H)，3135，3068υ(Ar-H)，2929，2867，υ(C-H)，1666，1635υ(C＝O)，1537，1306υ(N＝O)，1112δ(C-O)。

δ_H ¹H(DMSO)：2.42(4H，m，2(CH₂))，3.30(4H，q，(CH₂)-N-(CH₂)(J＝6.8Hz))，3.57(4H，t，(CH₂)-O-(CH₂)(J＝4.6Hz))，3.80(3H，s，N-Me)， 3.94(3H，s，N-Me)，6.82(1H，d，Ar-H(J＝1.6Hz))，7.20(1H，d，Ar-H(J＝1.6Hz))，7.57(1H，d，Ar-H(J＝1.6Hz))，7.93(1H，t，NH(J＝5.6Hz))，8.16(1H，d，Ar-H(J＝1.6Hz))，10.2(1H，s，NH)。

LREIMS: found 405.29(M + H) calcd for C₁₈H₂₄N₆O₅，404.18。

(v)4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } -carbonyl) -1H-pyrrol-3-yl]-1H-pyrrole-2-amides

1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrol-2-amide (150mg, 0.371 mmol; see step (iv) above) was suspended in methanol (25mL), to which was added Pd/C-10% (108mg) with stirring under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 5 hours. The catalyst was removed from the celite and the methanol was removed under reduced pressure to give the title compound which was used without further purification.

Preparation 2

4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]Amino } -N- [3- (dimethylamino) -propyl]-1-methyl-1H-pyrrole-2-amide

(i)N- [ 3-dimethylamino) propyl group]-1-methyl-4-nitro-1H-pyrrole-2-amide

(see, e.g., Abresia, n.g.a., Malinina, l., subira, j.a., j.mol.biol, 1999, 294, 657.)

A solution of 1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride (0.254g, 1.35 mmol; see preparation 1(ii) above) in DCM (10mL) was added dropwise over 15 minutes to DMPA (0.153g,. 5.05mmol) (Aldrich), NEt₃(0.152g, 1.5mmol) in DCM (10 mL). The resulting solution was stirred overnight before quenching the reaction with 5% NaOH solution (20 mL). The DCM fraction was then over MgSO₄Dried, filtered and the solvent removed under reduced pressure to yield the sub-title product (0.254g, 74%).

m.p.＝127-129℃，(lit.＝125-127℃)。

IR(KBr)：1498υ(C＝C)，1306，1532υ(N＝O)，1657υ(C＝O)，2794υ(CH₂)，2947υ(Ar-H)，3126υ(N-Me)，3284υ(N-H)cm^-1。

¹H NMR(DMSO)：1.62(2H，m，CH₂)，2.13(6H，s，2(CH₃))，2.24(2H，m，CH₂)，3.21(2H，m，CH₂)，3.90(3H，s，CH₃)，7.40(1H，d，Ar-H(J＝1.6Hz))，8.12(1H，d，Ar-H(J＝1.6Hz))，8.39(1H，t，NH(J＝5.6Hz))。

(ii)1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -N- [3- (dimethylamino) propyl group]-1H-pyrrole-2-amides

(see, e.g., Abresia, n.g.a., Malinina, l., subira, j.a., j.mol.biol, 1999, 294, 657.)

Reacting N- [3- (dimethylamino) propyl group]-1-methyl-4-nitro-1H-pyrrole-2-amide (0.241g, 0.95 mmol; see step (i) above) dissolved in methanol (10mL) and cooled to 0 deg.C, Pd/C (55mg) was added in small portions and the solution was in H₂Stirred for 3 hours. The solution was then filtered and the solvent removed under reduced pressure. A solution of 1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride (0.179g, 0.95 mmol; see preparation 1(ii) above) in DCM (10mL) was then added and stirred at room temperature for 1 hour, the solvent was removed under reduced pressure, and the crude product obtained was isolated on basified silica with a 1: 1 solution of methanolic ethyl acetate to yield the sub-title product (0.239g, 67%).

m.p.＝191-193℃，(lit.＝190-191℃)。

v_max KBr/cm^-1，1498υ(C＝C)，1537，1308υ(N＝O)，1621，1663υ(C＝O)，2821υ(CH₂)，2944υ(Ar-H)，3140υ(N-Me)，3287υ(N-H)。

δ_H ¹H(DMSO)，1.61(2H，m，CH₂)，2.14(6H，s，N(CH₃)₂)，2.25(2H，m，CH₂)，3.18(2H，m，CH₂)，6.81(1H，d，Ar-H(J＝1.6Hz))，7.20(1H，d，Ar-H(J＝1.6Hz))，7.56(1H，d，Ar-H(J＝1.6Hz))，8.11(1H，t，NH(J＝5.6Hz))，8.18(1H，d，Ar-H(J＝1.6Hz))，10.22(1H，s，NH)。

(iii)4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]amino-N- [3- (dimethylamino) propyl group]-1-methyl-1H-pyrrole-2-amide

1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrol-2-amide (150mg, 0.371 mmol; see step (ii) above) was suspended in methanol (25mL), to which was added Pd/C-10% (108mg) under stirring at 0 ℃ under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 5 hours. The catalyst was removed on celite and the methanol removed under reduced pressure to give the title compound which was used without further purification.

Preparation 3

2- { [ 4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]Amino } -N- [3- (dimethylamino) -propyl]-5-isopentyl-1, 3-thiazole-4-amide

(i)4-methylpentanal

To a vigorously stirred suspension of DCM (250mL) in pyridine chloro-chromium acid (Aldrich) (50g, 0.489mo1) was added over 45 minutes a solution of 4-methylpentanol (10g, 97.8mol) (Aldrich) in DCM (30 mL). The temperature was raised to 35 ℃ and the reaction mixture turned dark brown. After a total reaction time of 6 hours, ether (300mL) was added. The resulting brown solution was obtained by(50g, 30-60 mesh, Aldrich), and the precipitate from the ether was washed with an additional amount of ether (3X 30mL), and the solution was filtered as well. The resulting brown solution of 4-methylpentanal was concentrated to volume (350mL) and used in the next step without further purification or isolation.

(ii)Methyl 2-amino-5-isoamyl-1, 3-thiazole-4-carboxylate

(see, e.g., Wasserman, H.H.; Petersen, A.K. and Xia, M., Tetrahedron, 2003, 59, 6771-one 6784).

A solution prepared from Na (3g) and methanol (50mL, dry) was added over 45 minutes to a solution of methyl dichloroacetate (20g, 0.139mmol) and 4-methylpentanal (see step (i) above) and stirred vigorously at 0 ℃. After 1 hour at 0 deg.C, ether (50mL) and brine were added, the layers were collected and dried (MgSO)₄) The volatile solvent was removed under reduced pressure to yield a green liquid (16.40g) dissolved in methanol (60mL, dry) containing thiourea (8.50 g). The solution was heated at reflux for 4 hours, concentrated under reduced pressure and concentrated with 18M NH₄OH is neutralized. Extraction with DCM yielded the desired product as a light brown solid (13.50g, crude). This material was extracted with ethyl acetate/hexane (1/1R)_F0.15) through a column. Recrystallization of the product from acetone/hexane yielded pale yellow crystals (7.15g, 32%), m.p. ═ 108-. Recrystallization from petroleum ether (60-80) yielded the sub-title compound as white crystals.

¹H NMR(DMSO-d₆): 6.95(2H, s), 3.70(3H, s), 2.97(2H, t, J ═ 7.7Hz), 1.53(1H, heptad, J ═ 6.6Hz), 1.41(2H, q, J ═ 7.6Hz), 0.89(6H, d, J ═ 6.6 Hz).

(iii)Methyl 5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino) -1, 3-thiazole-4-carboxylic acid salts

1-methyl-4-nitro-1H-pyrrole-2-carboxylic acid (500mg, 2.94mmol) was suspended in thionyl chloride (5mL), and the reaction mixture was heated at reflux for 4 hours. Excess thionyl chloride was removed under reduced pressure at 50 ℃ and the acid chloride formed was dissolved in DCM (5mL, dry). Methyl 2-amino-5-isopentyl-1, 3-thiazole-4-carboxylate (728mg, 3.19 mmol; see step (ii) above) was dissolved in DCM (5mL, dry) and NMM (0.5mL, dry) was added with stirring at room temperature. The acid chloride solution is added dropwise to the amine at room temperature with stirringThe solution was stirred overnight. The reaction mixture was extracted with KOH solution (840mg in 10mL of water). The organic layer was extracted with brine and dried (MgSO)₄) The solvent was removed under reduced pressure. The crude product was eluted with silica gel and acetic acid/n-hexane (1/4), R_FPlace on column 0.20. The sub-title product was obtained as a white solid after crystallization from ethyl acetate/n-hexane (667mg, 60%), m.p. ═ 173-.

¹H NMR(DMSO-d₆): 12.81(1H, s), 8.30(1H, d, J ═ 1.6Hz), 7.99(1H, d, J ═ 1.6Hz), 3.97(3H, s), 3.80(3H, s), 3.31(2H, t, J ═ 7.7Hz), 1.58(1H, septematic, J ═ 6.6Hz), 1.53(2H, q, J ═ 7.6Hz), 0.92(6H, d, J ═ 6.6 Hz).

IR(KBr)：1720，1677，1561，1510，1423，1313，1230，1200，1112cm^-1。

HRFABMS: the following are found: 381.1223 calculation of value C₁₆H₂₁O₅N₄S 381.1233。

(iv)5-Isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -1, 3-thiazole-4-carboxylic acid

Methyl 5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -1, 3-thiazole-4-carboxylate (660mg, 1.74 mmol; see step (iii) above) was suspended in a mixture of water (25mL) and aluminum hydroxide (125mg, 5.21mmol) in methanol (5 mL). The reaction mixture was stirred vigorously at room temperature for 48 hours. Some of the methanol was removed at 50 ℃ under partial reduced pressure. The cooled solution was extracted with ether and the ether-containing layer was discarded. The aqueous layer was cooled to 0 ℃ and acidified by dropwise addition of HCl (concentrated) with stirring. The pale yellow solid was filtered off, washed with water and dried under reduced pressure at 45 ℃ overnight to give the sub-title compound (584mg, 92% yield), m.p. ═ 296-300 ℃.

¹H NMR(DMSO-d₆): 12.79(2H, br), 8.29(1H, d, J ═ 1.6Hz), 7.99(1H, d, J ═ 1.6Hz), 3.97(3H, s), 3.13(2H, t, J ═ 7.7Hz), 1.56(1H, septematic, J ═ 6.6Hz),1.53(2H，q，J＝7.6Hz)，0.92(6H，d，J＝6.6Hz)。

IR(KBr)：1669，1563，1514，1424，1315，1231，1200，1115cm^-1。

HRFABMS: the following are found: 367.1068 calculation of value C₁₅H₁₉O₅N₄S 367.1076。

(v)N- [3- (dimethylamino) propyl group]-5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -1, 3-thiazole-4-amides

5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -1, 3-thiazole-4-carboxylic acid (570mg, 1.56 mmol; see step (iv) above) was dissolved in DMF (2.5mL, dry), to which NMM (0.5mL, dry), HBTU (1.18g, 3.12mmol) and dimethylaminopropylamine (318mg, 3.12mmol, Aldrich) were added with stirring at room temperature. After standard work-up and purification, the sub-title compound was obtained as a yellow solid (660mg, 94%), m.p. > 230 ℃.

¹H NMR(DMSO-d₆): 8.31(1H, d, J ═ 1.6Hz), 7.96(1H, d, J ═ 1.6Hz), 3.98(3H, s), 3.33(2H, q, J ═ 6.4Hz), 3.19(2H, t, J ═ 7.7Hz), 2.62(3H, s), 1.84(2H, quintuple, J ═ 7.7Hz), 1.57(1H, septuple, J ═ 6.7Hz), 1.51(2H, q, J ═ 6.5Hz), 0.91(6H, d, J ═ 6.3).

IR(KBr)：1674，1642，1561，1502，1421，1310，1120cm^-1。

HRFABMS: the following are found: 451.2125 calculation of value C₂₀H₃₁O₄N₆S 451.2127。

(vi)2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]Amino } -N- (3- (dimethylamino) propyl group]-5-isopentyl-1, 3-thiazole-4-amide

N- [3- (dimethylamino) propyl ] -5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -1, 3-thiazole-4-amide (265mg, 0.588 mmol; see step (v) above) was suspended in methanol (25mL), to which Pd/C-10% (300mg) was added with stirring at 0 ℃ under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 3 hours. The catalyst was removed on celite and methanol removed under reduced pressure to give the title compound which was used without further purification.

Preparation 4

4- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid

(i)Methyl 4- [ (diethoxyphosphoryl) methyl group]Benzoic acid esters

(see, e.g., Tetrahedron, 2002, 58, 1425-

A mixture of methyl 4- (bromomethyl) benzoate (2.50g, 10.9mmol) and triethylphosphite (3.62g, 21.8mmol, 2 molar equivalents) was heated at 160 deg.C under nitrogen for 2 hours. Excess triethylphosphite was removed in vacuo to yield the subtitle compound as a colorless oil (3.03g, 97%).

¹H NMR(DMSO-d₆)：7.91(2H，d，J＝8.0Hz)，7.44(2H，dd，J＝2.4Hz &J ═ 8.4Hz), 4.00(4H, quintuple, J ═ 6.8Hz), 3.84(3H, s), 3.36(2H, d, J ═ 22.0Hz), 1.16(6H, t, J ═ 6.8 Hz).

(ii)Methyl 4- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid esters

Sodium hydride (0.678g, 60%, 18.8mmol) was added to methyl 4- [ (diethoxyphosphoryl) methyl group under nitrogen]Benzoate (3.03g, 10.6 mmol; see step (i) above) in THF (10mL, dry). After cooling the reaction mixture to 0 ℃, m-anisaldehyde (1.54g, 11.3mmol in THF 20mL, dry) was added carefully dropwise with stirring. The reaction mixture was stirred at room temperature for 1 hour and then quenched with water. After neutralization with dilute HCl, the two layers were separated. The aqueous layer was extracted with ethyl acetate, the organic layers were combined, dried (MgSO4), filtered and the solvent was removed under reduced pressure to yield a white solid. TLC displayShows two points (R)_F0.80 and R_F0.10, 20% ethyl acetate/n-hexane). The two compounds were separated by silica gel chromatography (20% ethyl acetate/n-hexane).

The first fraction was methyl 4- [ (E) -2- (3-methoxyphenyl) vinyl ] -benzoate isolated as a white solid (1.57g, 55%), m.p. ═ 92-94 ℃.

¹H NMR(DMSO-d₆)：7.96(2H，d，J＝8.4Hz)，7.74(2H，d，J＝8.4Hz)，7.42-7.21(4H，m)，6.88(1H，m)，3.85(3H，s)，3.80(3H，s)。

IR(KBr)：1708，1595，1438，1280，1244，1174，1105，1033，965，865，784，697cm^-1。

HREIMS: found 268.1100 calculated value C₁₇H₁₆O₃ 268.1099。

(iii)4- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid

In line with step (ii) above, the second fraction isolated as a white solid by silica gel chromatography (20% ethyl acetate/n-hexane) was 4- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoic acid (0.250g, 9%), m.p. -. 200-.

¹H NMR(DMSO-d₆)：12.88(1H，br)，7.94(2H，d，J＝8.4Hz)，7.72(2H，d，J＝8.4Hz)，7.37(2H，d，J＝3.6Hz)，7.31(1H，t，J＝8.0Hz)，7.22(2H，m)，6.88(1H，m)，3.80(3H，s)。

IR(KBr)：1674，1596，1429，1317，1280，1242，1180，1036，948，849，770cm^-1。

HREIMS: 254.0945 calculation of value C₁₆H₁₄O₃ 254.0977。

Preparation 5

4- [ (E) -2- (3-quinolyl) ethenyl]Benzoic acid

(i)Methyl 4- [ (E) -2- (3-quinolyl) ethenyl]Benzoic acid salts

Sodium hydride (0.678g, 60%, 18.8mmol) was added to methyl 4- [ (diethoxyphosphoryl) methyl group under nitrogen]A solution of benzoate (0.911g, 3.18 mmol; see preparation 4(i) above) in THF (10mL, dry). After cooling the reaction mixture to 0 ℃, quinoline-3-aldehyde (0.500g, 3.18mmol) in THF (10mL, dry) was added carefully dropwise with stirring. The reaction mixture was stirred at room temperature for 1 hour and quenched with water. After neutralization with dilute HCl, the two layers were separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined and dried (MgSO)₄) Filtration and removal of the solvent under reduced pressure gave a white solid. The product was taken up in 25% ethyl acetate/n-hexane (R)_FPurification on silica gel column (0.50) yielded the sub-title compound as a white solid (0.900g, 98%), m.p. ═ 92-94 ℃.

¹H NMR(DMSO-d₆)：9.25(2H，d，J＝2.0Hz)，8.54(2H，d，J＝2.0Hz)，8.01(2H，m)，7.82(2H，d，J＝8.4Hz)，7.75(1H，t，J＝6.8Hz)，7.64(2H，d，J＝3.6Hz)，7.63(1H，t，J＝8.1Hz)，3.87(3H，s)。

IR(KBr)：1716，1598，1460，1273，1173，750cm^-1。

HREIMS: the following are found: 289.1104 calculation of value C₁₉H₁₅NO₂ 289.1103。

(ii)4- [ (E) -2- (3-quinolyl) ethenyl]Benzoic acid

Methyl 4- [ (E) -2- (3-quinolyl) ethenyl ] benzoate (0.840mg, 3.36 mmol; step (i) above) was suspended in methanol (10mL) and water (20mL), to which was added, with stirring, sodium hydroxide solution (NaOH 0.580g, 14.5mmol in 10mL water). The reaction mixture was heated at reflux for 2 hours. At the beginning, the starting material dissolved and a white precipitate appeared. The reaction mixture was cooled in an ice bath and then diluted HCl was added dropwise with vigorous stirring until the pH was equal to 2. The title compound formed a yellow solid material, which was filtered, washed with water and dried in vacuo at 60 ℃ (0.640g, 69%), m.p. ═ 287 and 290 ℃.

¹H NMR(DMSO-d₆)：12.91(1H，br)，9.25(1H，d，J＝2.1Hz)，8.54(1H，d，J＝2.1Hz)，8.03-7.97(4H，m)，7.69-7.59(3H，m)。

IR(KBr)：1694，1586，1541，1423，1310，1272，1172，962，767，687cm^-1。

HREIMS: the following are found: 275.0948 calculation of value C₁₈H₁₃NO₂ 275.0946。

Preparation 6

4- [ (E) -2- (1-methyl-1H-pyrrol-2-yl) ethenyl]Benzoic acid, lithium salts

(i)Methyl 4- [ (E) -2- (1-methyl-1H-pyrrol-2-yl) ethenyl]Benzoic acid esters

To methyl 4- [ (diethoxyphosphoryl) methyl group in THF (10mL, dry)]To a solution of the benzoate (0.820g, 2.86 mmol; see preparation 4(i) above) was added sodium hydride (0.573g, 60%, 14.3mmol) under nitrogen. After cooling the reaction mixture to 0 ℃, N-methylpyrrole-2-aldehyde (0.312g, 2.86mmol) in THF (10mL, dry) was added dropwise with careful stirring. The reaction mixture was stirred at room temperature for 1 hour and quenched with water. After neutralization with dilute HCl, the two layers were separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure to yield a yellow solid. The product was taken up in 2% and gradually increased to 10% ethyl acetate/n-hexane (R)_F0.20) was purified by aluminium peroxide chromatography to give the sub-title compound as a yellow solid (0.140g, 20%), m.p. 95-98 ℃.

¹H NMR(DMSO-d₆)：7.91(2H，d，J＝8.4Hz)，7.68(2H，d，J＝8.0Hz)，7.35(1H，d，J＝16.4Hz)，6.95(1H，d，J＝16.4Hz)，6.83(1H，t，J＝2.0Hz)，6.56(1H，dd，J＝1.5Hz & J＝3.7Hz)，6.06(1H，t，J＝3.1Hz)，3.84(3H，s)，3.71(3H，s)。

IR(KBr)：1704，1597，1420，1270，1175，1107，955，767，745cm^-1。

HREIMS: the following are found: 241.1106 calculation of value C₁₅H₁₅NO₂ 241.1103。

(ii)4- [ (E) -2- (1-methyl-1H-pyrrole-2) ethenyl]Benzoic acid, lithium salts

Methyl 4- [ (E) -2- (1-methyl-1H-pyrrole-2) vinyl ] benzoate (40mg, 0.166 mmol; see step (i) above) was suspended in a mixture of methanol (1mL) and lithium hydroxide solution (16mg LiOH in 2mL water). The reaction mixture was heated at 60 ℃ with stirring overnight. This solution was lyophilized and the title compound was used without further purification.

Preparation 7

3- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid

(i)3- [ (diethoxyphosphoryl) methyl group]Benzoic acid salts

A mixture of methyl 3- (bromomethyl) benzoate (2.51g, 10.9mmol) and triethylphosphite (3.62g, 21.8mmol, 2 molar equivalents) was heated at 160 deg.C under nitrogen for 2 hours. Excess triethylphosphite was removed in vacuo to yield the subtitle compound as a colorless oil (3.03g, 97%).

¹H NMR(DMSO-d₆): 7.91(1H, s), 7.84(1H, d, J ═ 6.7Hz), 7.56(1H, d, J ═ 6.7Hz), 7.46(1H, t, J ═ 8.0Hz), 4.00(4H, quintuple, J ═ 6.8Hz), 3.85(3H, s), 3.36(2H, d, J ═ 22.0Hz), 1.16(6H, t, J ═ 6.8 Hz).

IR(KBr)：1722，1590，1442，1289，1251，1197，1103，1035，966，847，803，753cm^-1。

HREIMS: the following are found: 296.1179 calculation of value C₁₅H₂₁O₄P 296.1177。

(ii)Methyl-3- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid salts

To methyl 3- [ (diethoxyphosphoryl) methyl group]To a solution of benzoate (1.02g, 3.56 mmol; see step (i) above) in THF (5mL, dry) was added sodium hydride (0.212g, 60%, 18.8mmol) under nitrogen. After cooling the reaction mixture to 0 ℃, m-anisaldehyde (0.485g, 3.56mmol) dissolved in THF (10mL, dry) was added dropwise with careful stirring. The reaction mixture was stirred at room temperature for 1 hour and then quenched with water. After neutralization with dilute HCl, the two layers were separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. Silica gel chromatographic column for product (R)_F0.8020% ethyl acetate/n-hexane) gave the subtitle compound as a white solid (0.747g, 78%), m.p. ═ 92-94 ℃.

¹H NMR(DMSO-d₆)：8.16(1H，s)，7.91(1H，d，J＝8.1Hz)，7.86(1H，d，J＝8.1Hz)，7.55-7.20(6H，m)，6.87(1H，dd，J＝1.7Hz & J＝8.0Hz)，3.88(3H，s)，3.80(3H，s)。

IR(KBr)：1710，158，1467，1440，1267，1161，792，744，686cm^-1。

HREIMS: the following are found: 268.1101 calculation of value C₁₇H₁₆O₃ 268.1099。

(iii)3- [ (E) -2- (3-methoxyphenyl) vinyl]Benzoic acid

Methyl-3- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoate (0.740mg, 2.75 mmol; see step (ii) above) was suspended in methanol (10mL) and water (20mL), to which was added with stirring a lithium hydroxide solution (LiOH 0.199g, 8.27mmol in 10mL water). The reaction mixture was heated at reflux for 4 hours. The reaction mixture was cooled in an ice bath and then diluted HCl was added dropwise with vigorous stirring until the pH was equal to 2. The product was filtered as a white solid, washed with water and dried under vacuum at 60 ℃ to yield the title compound (0.235g, 34%) m.p. ═ 195-.

¹H NMR(DMSO-d₆)：8.12(1H，s)，7.78(1H，d，J＝7.6Hz)，7.63(1H，d，J＝7.6Hz)，7.37-7.18(7H，m)，6.83(1H，dd，J＝1.7Hz & J＝8.0Hz)，3.80(3H，s)。

IR(KBr)：1684，1586，1541，1423，1310，1272，961，767，687cm^-1。

HREIMS: the following are found: 254.0935 calculation of value C₁₆H₁₄O₃ 254.0943。

Preparation 8

1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl]-1H-pyrrole-2-carboxylic acid

(i)Diethyl 4-nitrobenzyl phosphate

A mixture of 4-nitrobenzyl bromide (2.05g, 0.949mmol) and triethyl phosphite (2.23g, 1.34mmol) was heated at 160 deg.C under nitrogen for 2 hours.

Excess triethylphosphite was removed in vacuo to yield the sub-title product as a brown oil (2.50g, 96%).

¹H NMR(DMSO-d₆)：8.20(2H，d，J＝8.1Hz)，7.57(2H，dd，J＝2.4Hz &J＝8.8Hz)，3.97(4H，q，J＝7.0Hz)，3.48(2H，d，J＝22.4Hz)，1.18(6H，t，J＝7.0Hz)。

IR(KBr)：2982，2910，1601，1521，1392，1347，1254，1028，959，864，777，695cm^-1。

HREIMS: the following are found: 273.0765 calculation of value C₁₁H₁₆O₅NP 273.0766。

(ii)Ethyl-1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl group]-1H-pyridinePyrrole-2-carboxylic acid salts

To a solution of diethyl 4-nitrobenzyl phosphate (0.525g, 1.92 mmol; see step (i) above) in THF (5mL, dry) under nitrogen was added sodium hydride (0.115g, 60%, 2.88 mmol). After cooling the reaction mixture to 0 ℃, ethyl 4-formyl-1-methyl-1H-pyrrole-2-carboxylate (0.348g, 1.92mmol) in THF (10mL, dry) was added dropwise with careful stirring. The reaction mixture was stirred at room temperature for 1 hour and then quenched with water. After neutralization with dilute HCl, the two layers were separated. The aqueous layer was extracted with ethyl acetate, and the organic layers were combined and dried (MgSO)₄) Filtered and the solvent removed under reduced pressure. The yellow solid was collected and triturated with warm methanol to give a yellow solid (288 mg). The mother liquor was collected and chromatographed on silica gel (R)_F0.5050% ethyl acetate/hexanes) to yield additional material (100 mg). The sub-title compound was isolated as a yellow solid (0.388g, 40%), m.p. -. 165-.

NMR(DMSO-d₆)：8.19(2H，d，J＝8.8Hz)，7.73(2H，d，J＝8.8Hz)，7.40(1H，d，J＝1.5Hz)，7.34(1H，d，J＝16.3Hz)，7.20(1H，d，J＝1.5Hz)，7.09(1H，d，J＝16.3Hz)，4.26(2H，q，J＝7.1Hz)，3.87(3H，s)，1.28(3H，t，J＝7.1Hz)。

IR(KBr)：1680，1632，1588，1546，1508，1338，1249，1142，1101，980，849，760cm^-1。

HREIMS: the following are found: 300.1111 calculation of value C₁₆H₁₆O₄N₂ 300.1110。

(iii)1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl]-1H-pyrrole-2-carboxylic acid

Ethyl-1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl]-1H-pyrrole-2-carboxylate (0.100mg, 0.333 mmol; see step (ii) above) was suspended in ethanol (4mL), THF (8mL) and water (20mL), to which was added with stirring a sodium hydroxide solution (NaOH 0.190g, 4.75mmol in 10mL water). The reaction mixture was heated at reflux for 4 hours. The reaction mixture was cooled to room temperature and thenAdding HCl dropwise with vigorous stirring_(Thick)Until the pH is equal to 2. The yellow solid was filtered, washed with water, and dried under vacuum at 60 ℃ to give the title compound (50mg, 55%), m.p. -. 212-.

¹H NMR(DMSO-d₆)：12.37(1H，br)，8.18(2H，d，J＝8.8Hz)，7.72(2H，d，J＝8.8Hz)，7.36(1H，d，J＝1.5Hz)，7.34(1H，d，J＝16.3Hz)，7.15(1H，d，J＝1.5Hz)，7.04(1H，d，J＝16.3Hz)，3.86(3H，s).

IR(KBr)：1671，1634，1588，1505，1449，1337，1281，1255，1184，1145，1105，833，802，742，688cm^-1。

HREIMS: the following are found: 272.0795 calculation of value C₁₄H₁₂O₄N₂ 272.0797。

Preparation 9

4- [ (E) -2- (4-pyridinyl) ethenyl]Phenyl acid

4-Carboxybenzaldehyde (2.03g, 13.3mmol, Aldrich) and 4-methylpyridine (1.24g, 13.3mmol, Aldrich) were placed in a round-bottomed flask, to which was added 20mL of anhydrous acetic acid. The reaction mixture was heated at reflux for 24 hours. The solid was filtered, washed with acetic acid and then water before drying in vacuo at 50 ℃ overnight to give the title compound as an off-white solid (426mg, 14%).

¹H NMR(DMSO-d₆)：7.40(1H，d，J＝16Hz)，7.60(2H，d，J＝6Hz)，7.62(1H，d，J＝16Hz)，7.77(2H，d，J＝8Hz)，7.97(2H，d，J＝8Hz)，8.57(2H，d，J＝6Hz)，10.10(1H，s)。

IR(KBr)：1606，1690，2995cm^-1。

HRFABMS: the following are found: 226.2424 calculation of value C₁₄H₁₁NO₂ 225.2426。

Preparation 10

Diethyl (1-methyl-4-nitro-1H-pyrrol-2-yl) methyl phosphate

(i)1-methyl-4-nitro-1H-pyrrole-2-carbaldehyde

(see, e.g., Suckling, c.j., Khalaf, a.i., Pitt a.r., Scobie, m., Tetrahedron, 2000, 56, 5225.)

Adding HNO₃(70%) (1.6mL) was added dropwise to anhydrous acetic acid (8mL) at-25 deg.C and stirred for a further 20 minutes. This solution was added dropwise to 1-methylpyrrole-2-carbaldehyde (Aldrich) (1.74g, 15.96mmol) in anhydrous acetic acid (12mL) at-25 ℃ and allowed to come to room temperature over 2 hours. The solution was cooled to-40 ℃ at which temperature a precipitate formed. The precipitate was collected, washed with hexane, and then dried under reduced pressure to give the sub-title compound (0.540g, 22%).

m.p.＝157-159℃，(Lit＝158-160℃)。

¹H NMR(CDCl₃)：4.04(3H，s，CH₃)，7.43(1H，d，Ar-H，J＝1.6Hz)，7.68(1H，d，Ar-H，J＝1.6Hz)。

IR(KBr)：3139υ(N-Me)，3125υ(Ar-H)，2958υ(C-H(COH))，1671υ(C＝O)，1504，1311υ(N＝O)cm^-1。

Analytical calculation C₆H₆O₃N₂C, 46.76; h, 3.92; n, 18.18; o, 31.14, found: c, 46.29; h, 3.68; n, 17.47.

(ii)(1-methyl-4-nitro-1H-pyrrol-2-yl) methanol

1-methyl-4-nitro-1H-pyrrole-2-carbaldehyde (0.400g, 2.08 mmol; see step (i) above) was placed under nitrogen in 50mL of anhydrous ethanol. Adding small amount of NaBH over 5 min₄(0.040g, 1.04mmol), and the solution was stirred for 20 minutes. The reaction was quenched by slow addition of water (10 mL). The organic was extracted with ethyl acetate (2X 20mL) to give an extractDrying of the fractions (MgSO)₄) Filtration, and removal of solvent under reduced pressure gave the sub-title compound as a pale brown solid (0.318g, 98%). m.p. ═ 89-90 ℃ and (Lit ═ 90.5-91.5 ℃).

IR(KBr)：3521υ(O-H)，3131υ(N-Me)，2934，2888υ(Ar-H)，1490，1412υ(C＝C)，1520，1337υ(N＝O)cm^-1。

¹H NMR(CDCl₃)：3.67(3H，s，N-Me)，4.40(2H，d，CH₂，J＝5.4Hz)，5.18(1H，t，OH，J＝3.0Hz)，6.57(1H，d，Ar-H，J＝1.6Hz))，7.92(1H，d，Ar-H，J＝1.6Hz)。

(iii)Diethyl (1-methyl-4-nitro-1H-pyrrol-2-yl) methylphosphonate

(1-methyl-4-nitro-1H-pyrrol-2-yl) methanol (0.100g, 0.64 mmol; see step (ii) above) was treated in DCM (5mL) and SOCl was slowly added₂(5mL), the solution was then refluxed for 15 minutes and excess SOCl was removed under reduced pressure₂. The residue was treated at 160 ℃ P (OEt)₃(3mL) for 1h, excess P (OEt) is removed under high vacuum (1.5mmHg at 70 ℃ C.)₃To afford the title compound (0.173g, 98%) as a brown oil which solidified to a brown crystalline solid after 48 hours at 0-4 ℃.

IR(NaCl)：3137υ(N-Me)，2985υ(Ar-H)，1556，1438υ(C＝C)，1519，1346υ(N＝O)，1308υ(P＝O)，1163δ(P-O-C)cm^-1。

¹H NMR(CDCl₃)：1.20(6H，t，CH₃，J＝6.8Hz)，3.08(2H，d，(CH₂)P，J＝20.4Hz)，3.65(3H，s，N-Me)，4.01(4H，q，(CH₂)CH₃，J＝6.8Hz)，6.54(1H，d，Ar-H，J＝1.6Hz)，7.39(1H，d，Ar-H，J＝1.6Hz)，δ_P δ³¹P(CDCl₃)，23.44.。

HRFABMS: found 276.0875 calculated value C₁₀H₁₇N₂O₅P 276.0873。

Preparation 11

Ethyl 4-formyl-1-methyl-1H-pyrrole-2-carboxylate

(i)2-trichloroacetyl-N-methylpyrrole

(see, e.g., Suckling, c.j., Khalaf, a.i., Pitt a.r., Scobie, m., Tetrahedron, 2000, 56, 5225.)

Trichloroacetyl (36.2g, 200.65mmol) in DCM (130mL) was placed in a round bottom flask under nitrogen at room temperature. A solution of N-methylpyrrole (16.2g, 200.32mmol) (Aldrich) in DCM (70mL) was added dropwise over 2.5 hours, and the solution was stirred overnight. The solvent was removed under reduced pressure to give the crude product, which was filtered through a silica gel column with DCM as eluent to give the sub-title product as a yellow solid (31.690g, 70%).

m.p.＝62-64℃，(Lit＝64-65℃)。

IR(KBr)：3137，3119υ(N-Me)，3005，2952υ(C-H)，1655υ(C＝O)，1238，1124υ(C-O)，742δ(C-H)cm^-1。

¹H NMR(CDCl₃)：3.98(3H，s，CH₃)，6.23(1H，dd，Ar-H，J＝2.6Hz &J＝3.9Hz)，5.97(1H，d，Ar-H，J＝1.6Hz)，7.51(1H，q，Ar-H，J＝7.2Hz)， ¹³C(CDCl₃)：38.68(CH₃)，96.52(CCl₃)，109.07(C)，122.02(C-H)，124.18(C-H)，133.80(C-H)，173.04(C＝O)。

(ii)Ethyl 1-methyl-1H-pyrrole-2-carboxylate

(see, e.g., Suckling, c.j., Khalaf, a.i., Pitt a.r., Scobie, m., Tetrahedron, 2000, 56, 5225.)

2-trichloroacetyl-N-methylpyrrole (1.47g, 6.51 mmol; see step (i) above) and EtOH (20mL) were placed in a round-bottomed flask, to which was added NaOEt (0.33g, 6.5)2mmol) and the resulting mixture was refluxed and heated with stirring for 1 hour. The reaction was quenched with water (10mL) and extracted with DCM (2X 20 mL). DCM fractions were combined and dried (MgSO)₄) Reduction in vacuo gave the sub-title compound (0.926g, 93%) as a yellow oil.

IR(KBr)，3136υ(N-Me)，2980υ(C-H)，1713υ(C＝O)，1244，1114υ(C-O)，599δ(C-H)cm^-1。

¹H NMR(CDCl₃)：1.35(3H，t，CH₂(CH₃)，J＝7.2Hz)，3.93(3H，s，CH₃)，4.27(2H，q，CH₂，J＝7.2Hz)，6.11(1H，dd，Ar-H，J＝2.6Hz & J＝4.0Hz)，6.78(1H，t，CH₂，J＝2.1Hz)，6.94(1H，dd，Ar-H，J＝2.6Hz & J＝4.0Hz)。

(iii)Ethyl 4-formyl-1-methyl-1H-pyrrole-2-carboxylate

Ethyl 1-methyl-1H-pyrrole-2-carboxylate (3.69g, 28.24 mmol; see step (ii) above) and AlCl₃(8.02g, 60.11mmol) was added to a solution of nitromethane (40mL) and 1, 2-dichloroethane (40mL) at-30 ℃. Methylene chloride methyl ether (2.5mL, 28mmol) in 1, 2-dichloroethane (10mL) was added quickly to the solution and the mixture was stirred at-30 ℃ for 16 h. The solution was poured over ice (50g) and the layers separated. The aqueous layer was then extracted with ether (50 mL). The combined organic fractions were dried (MgSO)₄) Filtration and removal of the solvent under reduced pressure gave the title compound as a crystalline brown/black solid (4.77g, 94%).

m.p.＝69-71℃，(Lit＝66-68℃)。

IR(NaCl)：3129υ(N-Me)，2981υ(Ar-H)，2767，2719υ(C-H(CHO))，1676υ(C＝O)，1541，1500，1471υ(C＝C)1260，1210υ(C-O)，1437δ(CH₃，CH₂)cm^-1。

¹H NMR(CDCl₃)：1.36(3H，t，CH₂(CH₃)，J＝7.2Hz)，3.98(3H，s，N-Me)，4.32(2H，q，(CH₂)CH₃，J＝7.2Hz)，7.37(2H，m，2(Ar-H))，9.76(1H，s，CO(H))。

Preparation 12

1-methyl-4- [ (E) -2- (3-quinolyl) ethenyl]-1H-pyrrole-2-carboxylic acid

(i)3-quinolylmethanol

3-Quinolinecarboxaldehyde (Aldrich) (1.04g, 6.64mmol) was dissolved in absolute ethanol (20mL) and NaBH was added in small amounts over 10 minutes₄(0.250g, 3.95mmol) and the resulting solution was stirred for an additional 30 minutes. Water (20mL) was then added and the resulting solution was extracted with ethyl acetate (2X 30 mL). The combined organics were dried (MgSO)₄) Filtration and removal of the solvent under reduced pressure gave the sub-title compound.

(ii)3- (chloromethyl) quinoline chloride

3-Quinolinemethanol (prepared using step (i) above) was dissolved in DCM (2mL) and the solution was added (initially dropwise) SOCl₂(5mL) and then refluxed for 1 hour. DCM and excess SOCl were then removed under reduced pressure₂To give the sub-title compound, which was used without further purification in the next step.

(iii)Diethyl 3-quinolinemethylcarbochosphates

3- (chloromethyl) quinoline chloride (prepared in step (ii) above) was dissolved in water (5mL) and washed with NaCO₃(1M) washed and the aqueous phase extracted with ethyl acetate (2X 15 mL). The combined organics were over MgSO₄Dried and the solvent removed under reduced pressure. The residue was then dissolved in POEt₃(3mL), the solution was refluxed for 1 hour. Removing excess POEt under high vacuum₃To give the sub-title compound as a viscous orange oil (1.816g, 98%).

v_max KBr/cm-1，3056，2982，2931，2907υ(C-H)，1606υ(C＝N)，1571，1495，1443υ(C＝C)，1253υ(P＝O)，1052δ(P-O-C)。

δ_H ¹H(CDCl₃)，1.25(6H，m，2(CH₃))，3.30(2H，d，CH₂(J＝21.9Hz))，4.06(4H，m，2(CH₂))，7.55(1H，t，Ar-H(J＝7.2Hz))，7.70(1H，t，Ar-H(J＝7.2Hz))，7.81(1H，d，Ar-H(J＝8.0Hz))，8.09(1H，d，Ar-H(J＝8.0Hz))，8.12(1H，s，Ar-H)，8.81(1H，s，Ar-H)，δ_P δ³¹P(CDCl₃)，25.77。

LREIMS: found 280.10(M + H) calculated value C₁₄H₁₈NO₃P 279.10。

(iv)Ethyl-1-methyl-4- [ (E) -2- (3-quinolyl) ethenyl]-1H-pyrrole-2-carboxylate

3-quinolinemethylcarbochosphate (0.503g, 1.93 mmol; see step (iii) above) was dissolved in anhydrous THF (2mL), NaH (0.273g, 11.37mmol) was added in small amounts to the solution, and the resulting mixture was aged for an additional 10 minutes. Ethyl 4-formyl-1-methyl-1H-pyrrole-2-carboxylate (0.313g, 1.92 mmol; see preparation 11 above) dissolved in anhydrous THF (3mL) was added dropwise to the solution and stirred for 16H. Water (5mL) was added to the mixture (initially dropwise) during which time the sub-title compound precipitated as a yellow solid (0.201g, 34%).

m.p.＝134-137℃。

v_max KBr/cm^-13001, 2924 υ (C-H), 2854 υ (N-Me)1699 υ (C ═ O), 1636 υ (C ═ C olefin), 16001546, 1494, 1436 υ (C ═ C), 1367 δ (C-H).

δ_H ¹H(CDCl₃)，1.30(3H，t，CH₃(J＝7.2Hz))，3.88(3H，s，N-CH₃)，4.25(2H，q，CH₂(J＝7.0Hz))，7.08(1H，d，C＝C(J＝16.4Hz))，7.19(1H，d，Ar-H(J＝1.6Hz))，7.35(2H，m，Ar-H)，7.58(1H，t，Ar-H(J＝7.2Hz))，7.68(1H，t，Ar-H(J＝7.2Hz))，.7.92(1H，d，Ar-H(J＝8.0Hz))，7.97(1H，d，Ar-H(J＝8.0Hz))，8.32(1H，s，Ar-H)，9.10(1H，d，Ar-H(J＝2.0Hz))。

HRFABMS: found 306.1371 calculated value C₁₉H₁₈N₂O₂ 306.1368。

(v)1-methyl-4- [ (E) -2- (3-quinolyl) ethenyl]-1H-pyrrole-2-carboxylic acid

Ethyl 1-methyl-4- [ (E) -2- (3-quinolyl) vinyl ] -1H-pyrrole-2-carboxylate (0.137g, 0.44 mmol; see step (iv) above) was suspended in ethanol (2mL), NaOH (0.052g, 1.32mmol) dissolved in water (5mL) was added to the solution, and the resulting mixture was refluxed for 2 hours. The reaction was filtered hot and then cooled to 0 ℃. Dilute HCl was added dropwise until the title compound precipitated as a yellow solid (0.076g, 62%).

m.p.＞230℃.

v_max KBr/cm^-1: 3462 υ (O-H), 2982 υ (N-Me), 2824 υ (Ar-H), 2854 υ (N-Me)1685 υ (C ═ O), 1639 υ (C ═ C olefin), 16031552, 1494 υ (C ═ C).

δ_H ¹H(DMSO)：3.86(3H，s，N-CH₃) 6.78(1H, d, Ar-H (J ═ 1.6Hz)), 6.88(1H, d, C ═ C olefin (J ═ 16.4Hz)), 6.97(1H, d, Ar-H (J ═ 1.6Hz)), 7.30(1H, d, C ═ C olefin (J ═ 16.4Hz)), 7.56(1H, t, Ar-H (J ═ 7.2Hz)), 7.65(1H, d, Ar-H (J ═ 7.2Hz)), 7.90(1H, d, Ar-H (J ═ 8.0Hz)), 7.95(1H, d, Ar-H (J ═ 8.0Hz)), 8.28(1H, s, Ar-H), 9.08(1H, s, Ar-H).

HRFABMS: found 278.1054 calculated value C₁₇H₁₄N₂O₂ 278.1055。

Preparation 13

1-methyl-4- [ (E) -2- (2-quinolyl) ethenyl]-1H-pyrrole-2-carboxylic acid

(i)Diethyl 2-quinolinemethyl-carbochosphates

The subtitle compound is prepared according to a similar procedure described above for preparation 12(iii) using 2- (chloromethyl) quinoline chloride instead of 3- (chloromethyl) quinoline chloride.

Yield 1.790g, 97%.

v_max KBr/cm^-1，3059，2982，2930，2907υ(C-H)，1618υ(C＝N)，1599，1562，1478，1442υ(C＝C)，1254υ(P＝O)，1027δ(P-O-C)。

δ_H ¹H(CDCl₃)，1.26(6H，m，2(CH₃))，3.67(2H，d，CH₂(J＝22.4Hz))，4.11(4H，m，2(CH₂))，7.52(2H，m，Ar-H)，7.70(1H，t，Ar-H(J＝7.2Hz))，7.81(1H，d，Ar-H(J＝8.0Hz))，8.05(1H，d，Ar-H(J＝8.0Hz))，8.12(1H，d，Ar-H(J＝8.0Hz))，δ_P δ³¹P(CDCl₃)，25.35。

HRFABMS: found 280.1098(M + H) calculated value C₁₄H₁₈NO₃P 279.1024。

(ii)Triphenyl (2-quinolylmethyl) phosphoryl chloride

2- (chloromethyl) phosphoryl chloride (0.669g, 1.52mmol) (Aldrich) was dissolved in water (5mL) and Na was added₂CO₃And (1M) washing. The aqueous phase was then extracted with ethyl acetate (2X 15 mL). The ethyl acetate extract was dried (MgSO)₄) The ethyl acetate was removed under reduced pressure. The residue was dissolved in toluene (10mL) and PPh₃(0.324g, 1.52mmol) the resulting solution was refluxed for 18 hours before cooling to 0 ℃. The precipitate formed was filtered and dried to yield the sub-title compound as a white solid.

Yield 0.468g, 31.3%, m.p. > 230 deg.c

v_max KBr/cm^-1，3053，2990，2964，2903υ(C-H)，1615υ(C＝N)，1591，1561，1485，1473υ(P-C)，1436υ(C＝C)。

δ_H ¹H(DMSO)，5.76(2H，d，CH₂(J＝15.1Hz))，7.5(1H，d，Ar-H，J＝8.4)，7.55(2H，t，Ar-H(J＝7.0Hz))，7.71(6H，m，Ar-H)，7.83(10H，m，Ar-H)，8.35(1H，d，Ar-H(J＝8.5Hz)).δ_P δ³¹P(CDCl₃)，25.35。

HRFABMS: discovery 404.1570 (M)⁺) Calculated value C₂₈H₂₃NP⁺ 404.1563。

(iii)Ethyl-1-methyl-4- [ (E) -2- (2-quinolyl) ethenyl]-1H-pyrrole-2-carboxylate

Alternative I

The subtitle compound was prepared according to a similar procedure to that described in preparation 12(iv) above, using diethyl 2-quinolinylmethyl phosphate (see step (i) above) instead of 3-quinolinylmethyl phosphate.

Yield 0.248g, 42%, m.p. 140-.

Substitute II

Triphenyl (2-quinolylmethyl) phosphoryl chloride (0.40g, 0.9 mmol; see step (ii) above) dissolved in THF (10mL) was placed in a round-bottomed flask at room temperature under nitrogen. BuLi (325. mu.L of 2.5M in hexane, 0.8mmol) was added dropwise with high stirring and the solution was stirred for an additional 1 hour. Ethyl 4-formyl-1-methyl-1H-pyrrole-2-carboxylate (0.175g, 0.9mmol) in THF (10mL) was then added dropwise over 10 minutes and the solution stirred overnight. The solvent was removed under reduced pressure and the residue was dry loaded onto silica and the product was eluted with 1: 2 ethyl acetate: hexane.

Yield 0.062g, 22%, m.p. 138-.

v_max KBr/cm^-1: 2997, 2894 υ (C-H), 2854 υ (N-Me)1689 υ (C ═ O), 1629 υ (C ═ C olefin) 1552, 1488, 1428 υ (C ═ C), 1359 δ (C-H).

δ_H ¹H(CDCl₃)，1.38(3H，t，2(CH₃)(J＝7.2Hz))，3.95(3H，s，N-CH₃)，4.31(2H，q，CH₂ J＝7.0Hz))，7.02(1H，d，Ar-H(J＝1.6Hz))，7.11(1H，d，(H)C＝CH(J＝16.4Hz))，7.28(1H，d，Ar-H(J＝1.6Hz))，7.48(1H，t，Ar-H(J＝7.2Hz))，7.55(2H，m，Ar-H)，7.72(1H，m，Ar-H)，7.88(1H，d，Ar-H(J＝8.0Hz))，8.08(2H，m，Ar-H)。

HRFABMS: found 306.1370 calculated value C₁₉H₁₈N₂O₂ 306.1368。

(iv)1-methyl-4- [ (E) -2- (2-quinolyl) ethenyl]-1H-pyrrole-2-carboxylic acid

The title compound was prepared according to a similar procedure described in preparation 12(v) above, using ethyl 1-methyl-4- [ (E) -2- (2-quinolinyl) vinyl ] -1H-pyrrole-2-carboxylate (see step (ii) above) in place of ethyl 1-methyl-4- [ (E) -2- (3-quinolinyl) vinyl ] -1H-pyrrole-2-carboxylate.

Yield 0.095g, 78%, m.p. > 230 ℃.

v_max KBr/cm^-1: 3423 υ (O-H)2978 υ (N-Me), 2822 υ (Ar-H), 2864 υ (N-Me)1680 υ (C ═ O), 1628 υ (C ═ C olefin), 15981554, 1489 υ (C ═ C).

δ_H ¹H(CDCl₃)：3.87(3H，s，N-CH₃) 6.84(1H, d, Ar-H (J ═ 1.6Hz)), 6.92(1H, d, C ═ C olefin (J ═ 16.4Hz)), 7.13(1H, d, Ar-H (J ═ 1.6Hz)), 7.48(1H, t, Ar-H (J ═ 7.2Hz)), 7.62(1H, d, C ═ C olefin (J ═ 16.4Hz)), 7.70(2H, m, Ar-H),. 7.89(2H, m, Ar-H), 8.22(1H, d, Ar-H (J ═ 8.0 Hz)).

HRFABMS: found 278.1054 calculated value C₁₇H₁₄N₂O₂ 278.1055。

Preparation 14

2- [ (E) -2- (2-quinolyl) ethenyl]-1, 3-thiazole-4-carboxylic acid

(i)2, 2-diEthoxyacetamide

(see, e.g., Inami, k., Shiba, t., bull. chem. soc. jpn., 1985, 58, 352.)

Ethyldiethoxyacetate (Aldrich) (10mL, 56.41mmol) was added to the concentrated NH₄In OH (50mL), the mixture was stirred until a homogeneous solution (40 h). The solvent was removed under reduced pressure to yield the fine white needle-like sub-title compound (8.209g, 99%).

m.p.＝47-51℃。

v_max KBr/cm^-1：3495，3298υ(N-H)，2898υ(C-H)，υ(N-Me)1673υ(C＝O)，1317δ(C-O)。

δ_H ¹H(CDCl₃)：1.11(6H，t，2(CH₃)(J＝8.0Hz))，4.55(4H，m，2(CH₂))，4.64(1H，s，C-H)，7.25(2H，d，NH₂(J＝188.0Hz))。

LREIMS: found 148.00(M + H) calculated value C₆H₁₃NO₃ 147.09。

(ii)Diethoxyacetonitrile

(see, e.g., Inami, k., Shiba, t., bull. chem. soc. jpn., 1985, 58, 352.)

2, 2-Dimethoxyacetamide (7.13g, 49.04 mmol; see step (i) above) was dissolved in toluene (50mL) and NEt was added₃(10.4mL) then P₂O₅(9.04g, 64.32mmol) and the mixture was stirred for 1 hour. The solvent was distilled off under normal pressure, and then the distillation was continued by placing the distiller under low pressure, and the subtitle compound (4.049g, 64%) was collected as a transparent colorless liquid.

B.p. 100 ℃ at 12-15 mmHg.

v_max KBr/cm^-1：2983，2937，2896υ(C-H)，2247υ(N≡C)，1067δ(C-O)。

δ_H ¹H(CDCl₃)：1.27(6H，t，2(CH₃)(J＝8.0Hz))，3.73(4H，m，2(CH₂))，5.31(1H，s，C-H)。

LREIMS: found 130.13(M + H) calculated value C₆H₁₁NO₂ 129.08。

(iii)2, 2-diethoxyethanethiolamine

(see, e.g., Inami, k., Shiba, t., bull. chem. soc. jpn., 1985, 58, 352.)

Diethoxyacetonitrile (0.050g, 0.5 mmol; see step (ii) above) was placed in a 10mL microwave oven vial, followed by addition of methanol (5mL) and (NH)₄)₂S (54. mu.L, 0.5mmol (40% wt aqueous solution)). The vial was heated at 100W, 80 ℃ for 15 minutes and then cooled to room temperature. The solvent was then removed under reduced pressure and the residue was dissolved in ethyl acetate (15mL) and extracted with water (2X 10mL) and brine (2X 5 mL). The organic layer was then dried (MgSO₄) Filtration and concentration under reduced pressure gave the sub-title compound as a white/off-white solid (0.081g, 99%).

m.p.＝92-94℃。

v_max KBr/cm^-1：3335，3171υ(N-H)，2976，2885υ(C-H)，2247υ(N≡C)，1645，1449υ(C＝S)，1019δ(C-O)。

δ_H ¹H(CDCl₃)：1.26(6H，t，2(CH₃)(J＝8.0Hz))，3.71(4H，m，2(CH₂))，5.05(1H，s，C-H)，7.70(2H，d，N-H)。

(iv)Ethyl 2- (diethoxymethyl) -1, 3-thiazole-4-carboxylate

(see, e.g., Inami, k., Shiba, t., bull. chem. soc. jpn., 1985, 58, 352.)

2, 2-Diethoxyethylthioamine (0.654g, 6.03 mmol; see step (iii) above) and ethylbromopyruvate (Aldrich) (1.28g, 6.10mmol) in 4Molecular sieves were dissolved in ethanol (10mL) in the presence of molecular sieves. The mixture was refluxed for 45 minutes and the solvent was removed under reduced pressure. The residue was dissolved in ethyl acetate (20mL) and taken up with saturated NaHCO₃The solution (2X 20mL) and brine (2X 20mL) were extracted. Then dried (MgSO)₄) The organic portion was filtered and the solvent removed under reduced pressure to yield the sub-title compound (1.452g, 93%).

v_max KBr/cm^-1：3108υ(Ar-H)，2981，2885，2856υ(C-H)，1735υ(C＝O)，1505υ(C＝N)，1042δ(C-O)。

δ_H ¹H(CDCl₃)：1.29(9H，m，2(CH₃))，3.71(4H，m，2(CH₂))，4.47(2H，q，CH₂(J＝7.8Hz))，5.70(1H，s，C-H)，8.19(1H，s，Ar-H)。

HRFABMS: found 260.0956(M + H) calculated value C₁₁H₁₇NO₄S 259.0878。

(v)Ethyl 2-formyl-1, 3-thiazole-4-carboxylic acid salt

(see, e.g., Inami, k., Shiba, t., bull. chem. soc. jpn., 1985, 58, 352.)

Ethyl 2- (diethoxymethyl) -1, 3-thiazole-4-carboxylate (1.34g, 5.17 mmol; see step (iv) above) was treated (take up) in acetone (100mL) and then 1N HCl (12.8mL) was added. The solution was refluxed for 45 minutes and the solvent was removed under reduced pressure. The residue was then dissolved in ethyl acetate (40mL) and saturated NaHCO₃The solution (2X 40mL) and brine (2X 40mL) were extracted. Then dried (MgSO)₄) The organic portion was filtered and the solvent removed under reduced pressure to yield the sub-title compound as a light brown solid (0.937g, 98%).

m.p.＝65-67℃(Lit＝67-68℃)。

v_max KBr/cm^-1：3116υ(Ar-H)，2983，2910，2814υ(C-H)，173υ(C＝O)，1513υ(C＝N)，1060δ(C-O)。

δ_H ¹H(CDCl₃)：1.47(3H，t，CH₃(J＝8.0Hz))，4.50(2H，q，CH₂(J＝8.0Hz))，8.52(1H，d，Ar-H(J＝1.2Hz))，10.08(1H，d，Ar-COH(J＝1.2Hz))。

HRFABMS: found 186.0228(M + H) calculated value C₇H₇NO₃S 185.0147。

(vi)Ethyl 2- [ (E) -2- (2-quinolyl) ethenyl]-1, 3-thiazole-4-carboxylic acid salt

Diethyl 2-quinolinemethylphosphate (0.580g, 2.08 mmol; see preparation 13(i) above) was dissolved in anhydrous THF (2mL) and NaH (0.273g, 11mmol) was added in small amounts and the resulting solution was stirred for 10 min. Ethyl 2-formyl-1, 3-thiazole-4-carboxylate (0.387g, 2.08 mmol; see step (v) above) in anhydrous THF (3mL) was added dropwise and the solution stirred for 16 hours. Water (5mL) was then added (initially dropwise) over this time, during which time the sub-title compound precipitated as a light brown/yellow solid.

Yield 0.232g, 36%, m.p. 183-186 ℃.

v_max KBr/cm^-1: 3124, 3043, 2955, 2925, 2899, 2853, 1730, 1612, 1627, 15921553, 1479, etc.

δ_H ¹H(CDCl₃)：1.34(3H，t，CH₃(J＝8.0Hz))，4.34(2H，q，CH₂(J ═ 8.0Hz)), 7.62(1H, t, Ar-H (J ═ 7.2Hz)), 7.74(1H, d, C ═ C olefin (J ═ 16.1Hz)), 7.79(1H, t, Ar-H (J ═ 6.8Hz)), 7.98(3H, m, Ar-H), 8.05(1H, d, C ═ C olefin (J ═ 16.1Hz)), 8.42(1H, d, Ar-H (J ═ 8.5Hz)), 8.56(1H, s, Ar-H).

HRFABMS: found 310.0772 calculated value C₁₇H₁₄N₂O₂S 310.0776。

(vii)2-[(E)-2-(2-quinolyl) ethenyl]-1, 3-thiazole-4-carboxylic acid

2- [ (E) -2- (2-quinolyl) vinyl ] -1, 3-thiazole-4-carboxylic acid salt (0.137g, 0.44 mmol; see step (vi) above) was suspended in ethanol (2mL), NaOH (0.052g, 1.32mmol) in water (5mL) was added, and the solution was refluxed for 2 hours. The reaction was then filtered while hot and subsequently cooled to 0 ℃. Diluted HCl was then added dropwise until the title compound precipitated as a yellow solid (0.103g, 83%).

m.p.＝218-220℃。

v_max KBr/cm^-1: 3469 upsilon (O-H), 3105, 3068 upsilon (Ar-H), 2923, 2853, upsilon (C-H), 1715 upsilon (C ═ O), 1640, 1632 upsilon (C ═ C olefin), 15991541, 1493 upsilon (C ═ C), 1320 δ (C-O).

δ_H ¹H(CDCl₃): 7.74(1H, t, Ar-H (J ═ 7.2Hz)), 7.76(1H, d, C ═ C olefin (J ═ 16.1Hz)), 7.93(1H, t, Ar-H (J ═ 7.2Hz)), 8.11(1H, d, Ar-H (J ═ 8.5Hz)), 8.05(1H, d, Ar-H (J ═ 8.5Hz)), 8.19(2H, m, Ar-H), 8.24(1H, d, C ═ C olefin (J ═ 16.1Hz)), 8.58(1H, s, Ar-H), 8.65(1H, d, Ar-H (J ═ 8.5 Hz)).

HRFABMS: found 282.0465 calculated value C₁₇H₁₄N₂O₂ 282.0463。

Preparation 15

4- [ (E) -2- (2-quinolyl) ethenyl]Benzoic acid

To a solution of methyl 4- [ (diethoxyphosphoryl) methyl ] carbonyl in THF (20mL, dry)]Benzoate (910mg, 3.18 mmol; see preparation 4(i) above) sodium hydride (520mg, 60% dispersion in oil, 13.00mmol) was added at 0 ℃ under nitrogen. 2-Quinolinecarboxaldehyde (500mg, 3.18mmol) was then dissolved in THF (10mL, dry) and added dropwise to the reaction mixture at 0 deg.C under nitrogen. The reaction mixture was stirred at room temperature for 1 hour and then quenched with water. The pH level of the mixture was added HCl dropwise with stirring at 0 deg.C_(Thick)Adjusting to pH4. The resulting yellow solid material was collected by filtration and suspended in methanol (5mL), to which was added sodium hydroxide (572mg, 10mL of water). The reaction mixture was heated at reflux for 3 hours and a yellow solid formed. The reaction mixture was extracted with ether (100mL) and the solid dissolved. The water layer was collected and cooled to 0 ℃, concentrated HCl was added dropwise with stirring until pH 4, at which time a precipitate formed. The yellow precipitate was collected, washed with water and dried to obtain the title compound (403mg, 46%, carboxylic acid), m.p.: 265-.

¹H NMR(DMSO-d₆)：13.01(1H，br)，8.50(1H，d，J＝8.5Hz)，8.07(1H，d，J＝8.5Hz)，8.02-7.96(5H，m)，7.87(2H，d，J＝8.4Hz)，7.84(1H，t，J＝7.2Hz)，7.67-7.61(d，J＝16.5Hz & t，J＝6.5Hz)。

IR(KBr)：3383，2593，1716，1628，1603，1416，1324，1209，1105，970，845，747cm^-1。

HRCIMS: the following are found: 276.1022 calculation of value C₁₈H₁₄O₂N 276.1025。

Preparation 16

4- [ (E) -2- (2-chloro-3-quinolinyl) ethenyl]Benzoic acid

The title compound was prepared according to a similar procedure described in preparation 15 above, substituting 2-chloro-3-quinolinecarboxaldehyde for 2-quinolinecarboxaldehyde to give the title compound as a white solid (495mg, 61%), m.p. ═ 190 ℃ (softened) then sublimed at about 270 ℃.

¹H NMR(DMSO-d₆)：13.00(1H，br)，8.92(1H，s)，8.08(1H，d，J＝7.6Hz)，8.03-7.95(3H，m)，7.84-7.77(3H，m)，7.70-7.55(3H，m)。

IR(KBr)：3383，2593，1716，1628，1603，1416，1324，1209，747cm^-1。

HREIMS: found 309.0558 calculated value C₁₈H₁₂ ³⁵ClNO₂ 309.0556。

Preparation 17

4- [ (E) -2- (2-naphthyl) ethenyl]Benzoic acid sodium salt or sodium 4- ((E) -2- (naphthalen-6-yl) vinyl) benzoate

(i)Methyl 4- [ (E) -2- (2-naphthyl) ethenyl]Benzoic acid salts

To methyl 4- [ (diethoxyphosphoryl) methyl group]To a solution of benzoate (916mg, 3.20 mmol; see preparation 4(i) above) in THF (20mL, dry) was added sodium hydride (520mg, 60% dispersion in oil, 13.00mmol) at 0 deg.C under nitrogen. 2-Naphthaldehyde (500mg, 3.20mmol) was dissolved in THF (10mL, dry) and the reaction mixture was added dropwise at 0 deg.C under nitrogen and stirred at room temperature for 1 hour followed by quenching with water. The pH level of the mixture was added HCl dropwise with stirring at 0 deg.C_(Thick)The pH was adjusted to 4. TLC analysis indicated it to be the desired product (282 mg). The aqueous layer was extracted with ethyl acetate and then dried (MgSO)₄) The sub-title compound (total 846mg, 92%) was produced, m.p. ═ 195 ℃ 198 ℃.

¹H NMR(DMSO-d₆)：8.07(1H，s)，7.99(2H，d，J＝8.4Hz)，7.95-7.90(3H，d & m，J＝6.3Hz)，7.81(2H，d，J＝8.4Hz)，7.63(1H，d，J＝16.4Hz)，7.55-7.48(3H，d & m，J＝16.4Hz)，3.87(3H，s)。

IR(KBr)：1718，1601，1455，1439，1411，1284，1193，1179，1110，962，869，819，748，700cm^-1。

HREIMS: found 288.1151 calculated value C₂₀H₁₆O₂ 288.1150。

(ii)4- [ (E) -2- (2-naphthyl) ethenyl]Benzoic acid sodium salt or sodium 4- ((E) -2- (naphthalen-6-yl) vinyl) benzoate

Methyl 4- [ (E) -2- (2-naphthyl) vinyl ] benzoate (280mg, 0.972 mmol; see step (i) above) was suspended in methanol (2mL) and sodium hydroxide solution (NaOH550mg, 13.75mmol in 10mL water) was added. The reaction mixture was stirred at reflux overnight and heated. The solvent was removed under reduced pressure and the resulting white solid was triturated with water and filtered to give the title compound (285mg, 99%).

m.p.＞230℃。

¹H NMR(DMSO-d₆)：8.15-7.75(8H，m)，7.60-7.35(5H，m)。

IR(KBr)：3393，1717，1583，1535，1418，1283，1102，957，865，824，741cm^-1。

HREIMS: found 229.1015 calculated value C₁₈H₁₃229.1017 (decarboxylation occurs in the probe).

Preparation 18

2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]Amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl-1, 3-thiazole-4-carboxamide

(i)5-Isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-carboxamide

Coupling 5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -1, 3-thiazole-4-carboxylic acid (330mg, 0.90 mmol; see preparation 3(iv) above) was dissolved in DMF (2.5mL, dry), to which NMM (0.1mL, dry), 2- (4-morpholinyl) ethylamine (117mg, 0.90mmol) and HBTU (682mg, 1.80mmol) were added with stirring at room temperature. Stirring was continued at room temperature for 24 h, then the reaction mixture was extracted with sodium hydroxide solution (381mg, 9.52mmol in 10mL water) and ethyl acetate (3X 50 mL). The organic layer was collected and dried (MgSO)₄) The solvent was removed under reduced pressure. The crude product was eluted with 1% TEA in ethyl acetate/methanol (R)_F0.15) was purified by chromatography column. The sub-title compound was obtained as a light yellow solid (400mg, 93%).

m.p.＝189-192℃。

¹H NMR(DMSO-d₆)：12.48(1H，br)，8.30(1H，d，J＝1.6Hz)，7.95(1H，d，J＝1.6Hz)，7.66(1H，t，J＝5.6Hz)，3.98(3H，s)，3.59(4H，t，J＝4.4Hz)，3.40(2H，q，J＝6.2Hz)，3.20(2H，t，J＝7.7Hz)，2.69(3H，s)，2.45-2-40(4H，m)，1.61-1.48(3H，m)，0.92(6H，d，J＝6.4Hz)。

IR(KBr)：3404，1661，1559，1474，1310，1292，1110，841cm^-1。

HRFABMS: the following are found: 479.2074 calculation of value C₂₁H₃₁O₅N₆S 479.2077。

(ii)2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl]Amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-carboxamide

5-isopentyl-2- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (400mg, 0.836 mmol; see step (i) above)) was dissolved in methanol (25mL), to which was added Pd/C-10% (300mg) under stirring at 0 ℃ under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 4 hours to give the title compound, which was used without further purification.

Preparation 19

6- ((E) -2- [ 4-methylthio) phenyl]Ethyl nicotinic acid

(i)Methyl 6- ((E) -2- [4- (methylthio) phenyl]Ethyl) } nicotinate

A mixture of 4- (methylthio) benzaldehyde (250mg, 1.52mmol), methyl 6-methylnicotinate (230mg, 1.52mmol), anhydrous acetic acid (310mg, 3.04mmol) and a catalytic amount of zinc chloride was heated at reflux for 12 hours. The reaction mixture was cooled to room temperature until the mixture was basic (pH 8) before crushed ice and then 10% (stirring and dropping) of sodium hydroxide solution were added. Dichloromethane ice was added and the layers were separated. The combined organic layers were collected and dried (MgSO)₄) Concentration under reduced pressure gave the crude product as a brown solid. The crude mixture was purified by column chromatography (ethyl acetate/n-hexane, 1: 1, R)_F0.80), followed by recrystallization from methanol to give the sub-title compound as pale yellow crystals (251mg, 58%).

m.p.＝157-159℃。

¹H NMR(DMSO-d₆)：9.04(1H，d，J＝2.0Hz)，8.26(1H，dd，J＝2.2Hz &J＝8.2Hz)，7.81(1H，d，J＝16.0Hz)，7.65(3H，d，J＝8.3Hz)，7.38(1H，d，J＝16.0Hz)，7.29(2H，d，J＝8.4Hz)，3.15(3H，s)，2.49(3H，s)。

IR(KBr)：1712，1586，1435，1280，1114，978，849，813，774，733cm^-1。

HRCIMS: the following are found: 286.0899 calculation of value C₁₆H₁₆O₂NS 286.0902。

(ii)6- { (E) -2- [4- (methylthio) phenyl]Vinyl nicotinic acid

Reacting methyl 6- { (E) -2- [4- (methylthio) phenyl]Vinyl nicotinate (80mg, 0.280 mmol; see step (i) above) was suspended in methanol (4mL) and sodium hydroxide solution (250mg in 10mL of water), and the reaction mixture was heated at reflux for 4 hours. The solvent was removed under reduced pressure and the aqueous layer was extracted with DCM. The aqueous layer was ice cooled to 0 ℃ and HCl was added dropwise with stirring_(Thick)Until a pH of 4 was reached, a precipitate formed. The solid was filtered, washed with water and dried under reduced pressure to give the title compound as a light yellow solid (75mg, 99%).

m.p.＝250-253℃.

¹H NMR(DMSO-d₆)：9.02(1H，d，J＝2.0Hz)，8.23(1H，dd，J＝2.2Hz &J＝8.2Hz)，7.79(1H，d，J＝16.0Hz)，7.63(3H，m)，7.37(1H，d，J＝16.0Hz)，7.29(2H，d，J＝8.4Hz)，4.49(3H，s)。

IR(KBr)：3419，2920，2500，1698，1565，1493，1381，1287，1142，1090，967，809，775，786cm^-1。

HRFABMS discovery: 272.0739 calculation of value C₁₅H₁₄O₂NS 272.0745。

Preparation 20

2- { (E) -2- [4- (methylthio) phenyl]Ethyl } | -6-quinolinecarboxylic acid

A mixture of 4- (methylthio) benzaldehyde (220mg, 1.33mmol), 2-methyl-6-quinolinecarboxylic acid (250mg, 1.33mmol), anhydrous acetic acid (310mg, 3.04mmol), zinc chloride (catalytic amount) and xylene (1mL) was heated at 140 ℃ overnight. The yellow solid precipitated and was triturated with water and ethyl acetate containing 5% methanol and filtered. The title compound was dried under reduced pressure at 50 ℃ overnight and yielded a yellow solid (270mg, 63%).

294-_F0.50 (ethyl acetate, fluorescent under UV lamp).

¹H NMR(DMSO-d₆)：13.12(1H，br)，8.61(1H，d，J＝1.8Hz)，8.53(1H，d，J＝8.6Hz)，8.19(1H，dd，J＝1.8Hz & J＝8.7Hz)，8.03(8.7Hz)，7.93(1H，d，J＝8.6Hz)，7.90(1H，d，J＝16.2Hz)，7.72(2H，d，J＝8.4Hz)，7.49(1H，d，J＝16.3Hz)，7.33(2H，J＝8.4Hz)，2.52(3H，s)。

IR(KBr)：3422，2918，2534，1681，1615，1586，1493，1472，1290，1185，1089，960，820，756cm^-1。

HRCIMS: the following are found: 322.0905 calculation of value C₁₉H₁₆O₂NS 322.0902。

Preparation 21

6- [ (E) -2- (4-methoxyphenyl) vinyl]Nicotinic acid

(i)Methyl 6- [ (E) -2- (4-methoxybenzene)Radical) vinyl]Nicotinate salt

4-Methoxybenzaldehyde (210mg, 1.52mmol), methyl 6-methylnicotinate (230mg, 1.52mmol), anhydrous acetic acid (310mg, 3.04mmol) and a catalytic amount of zinc chloride were heated at 140 ℃ with stirring for 12 hours. To the cooled reaction mixture were added ethyl acetate and brine, and the product was extracted. The combined organic layers were dried (MgSO)₄) The solvent was removed under reduced pressure. The crude product was placed on a silica gel column and the product was eluted with ethyl acetate/n-hexane (1: 1R)_FFluorescence under UV lamp 0.60). Fractions containing the desired compound were collected and the solvent was removed under reduced pressure to give the sub-title compound as a yellow solid (87mg, 21%).

170 ℃ [ reference m.p.: 170 ℃: cluzan, r.and Katz, l.the books Company ltd. U.S. Pat. No. 4,009,174, 1977 ].

¹H NMR(DMSO-d₆)：9.04(1H，d，J＝2.0Hz)，8.25(1H，dd，J＝2.2Hz &J＝8.2Hz)，7.81(1H，d，J＝16.0Hz)，7.67-7.62(3H，m)，7.28(1H，d，J＝16.0Hz)，7.00(2H，d，J＝8.8Hz)，3.88(3H，s)，3.80(3H，s)。

IR(KBr)：1717，1606，1591，1511，1433，1290，1254，1175，1111，1020，844，818，760，734cm^-1。

HRCIMS: the following are found: 270.1127 calculation of value C₁₆H₁₆O₃N 270.1130。

(ii)6- [ (E) -2- (4-methoxyphenyl) vinyl]Nicotinic acid

Methyl 6- [ (E) -2- (4-methoxyphenyl) vinyl ] nicotinate (80mg, 0.297 mmol; see step (i) above) was dissolved in methanol (5mL), to which was added sodium hydroxide solution (145mg in 10mL water). The reaction mixture was heated at reflux for 3 hours. The solvent was removed under reduced pressure and the remaining solution was cooled to 0 ℃. Hydrochloric acid (concentrated) was added dropwise with vigorous stirring until pH 4 and a precipitate formed. The yellow solid was collected by filtration, washed with water and dried under reduced pressure at 50 ℃ to give the title compound (59mg, 78%).

233 ℃ (about 200 ℃ sublimation) 230-.

¹H NMR(DMSO-d₆)：9.03(1H，d，J＝2.0Hz)，8.29(1H，dd，J＝2.1Hz &J＝8.2Hz)，7.90-7.65(4H，m)，7.29(1H，d，J＝16.0Hz)，7.01(2H，m)，3.80(3H，s)。

IR(KBr)：1717，1681，1635，1595，1513，1292，1250，1173，1023，825cm^-1。

HRFABMS: the following are found: 256.0972 calculation of value C₁₅H₁₄O₃N 256.0974。

Preparation 22

2- [ (E) -2- (4-methoxyphenyl) vinyl]-6-quinolinecarboxylic acids

A mixture of 2-methyl-6-quinolinecarboxylic acid (200mg, 0.936mmol), 4-methoxybenzaldehyde (228mg, 0.936mmol), anhydrous acetic acid (630mg, 6.18mmol) and a catalytic amount of zinc chloride was heated at 140 ℃ with stirring for 12 hours. The cooled reaction mixture was extracted with water and ethyl acetate. The organic layer was collected and dried (MgSO)₄) The solvent was removed under reduced pressure to yield a dark brown solid. The material was placed on a silica gel column and the product was eluted with ethyl acetate/n-hexane (1/4). Collecting the fractions containing the desired product (R)_F0.20 ethyl acetate/n-hexane 2/1 with fluorescence under UV lamp), the solvent was removed under reduced pressure. The residue as a yellow solid was triturated with n-hexane and filtered to give the title compound (71mg, 25%), m.p. -. 274-.

¹H NMR(DMSO-d₆)：13.11(1H，br)，8.60(1H，d，J＝1.8Hz)，8.51(1H，d，J＝8.6Hz)，8.19(1H，dd，J＝1.9Hz & J＝8.8Hz)，7.91(1H，d，J＝8.7Hz)，7.89(1H，d，J＝16.4Hz)，7.72(2H，d，8.8Hz)，7.38(1H，d，J＝16.3Hz)，7.02(2H，d，J＝8.8Hz)，3.81(3H，s)。

IR(KBr)：1682，1615，1593，1510，1467，1289，1252，1170，1030，968，830cm^-1。

Hreims. discovery: 305.1055 calculation of value C₁₉H₁₅NO₃ 305.1052。

Preparation 23

1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino) -carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-4-amino-1H-pyrrole-2-amides

(i)1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino) -carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-4-nitro-1H-pyrrole-2-amides

Coupling 1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl]Amino } -N- [2- (4-morpholinyl) ethyl]-1H-pyrrole-2-amide (0.343g, 0.85 mmol; see preparation 1(iv) above) dissolved in methanol (10mL) and cooled to 0 deg.C, Pd/C (200mg) was added in small amounts and the solution was in H₂Stirred for 3 hours. The solution was then filtered and the solvent removed under reduced pressure. A solution of 1-methyl-4-nitro-1H-pyrrole-2-carbonyl chloride (160g, 0.85mmol) in DCM (10mL) was added, stirred at room temperature for 1 hour, the solvent was removed under reduced pressure, and the crude product obtained was isolated with a solution of 1: 1 ethyl methylacetate on basified silica to give the sub-title compound.

Yield 0.259g, 58%, m.p. > 230 ℃.

v_max KBr/cm^-1：3334，3279υ(N-H)，3135，3068υ(Ar-H)，2929，2867，υ(C-H)，1671，1633υ(C＝O)，1535，1308υ(N＝O)，1115δ(C-O)。

δ_H ¹H(DMSO)：3.13(2H，m，CH₂)，3.25(2H，m，CH₂)，3.58(4H，m，(CH₂)-O-(CH₂))，3.74(2H，m，CH₂)，3.82(3H，s，N-Me)，3.86(3H，s，N-Me)，3.96(3H，s，N-Me)，6.98(1H，d，Ar-H(J＝1.6Hz))，7.05(1H，d，Ar-H(J＝1.6Hz))，7.22(1H，d，Ar-H(J＝1.6Hz))，7.26(1H，d，Ar-H(J＝1.6Hz))，7.60(1H，d，Ar-H(J＝1.6Hz))，8.18(1H，d，Ar-H(J＝1.6Hz))， 8.28(1H，t，N-H(J＝5.6Hz))，9.97(1H，s，NH)，10.12(1H，s，NH)。

LREIMS: found 527.42(M + H) calculated value C₂₄H₃₀N₈O₆ 526.25。

(ii)1-methyl-N- [ 1-methyl-5- ({ [ 1-methyl-5- { ([ 2-4-morpholinyl)]Ethyl } amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-4-amino-IH-pyrrole-2-amide

1-methyl-N- [ 1-methyl-5- ({ [ 1-methyl-5- { ([ 2-4-morpholinyl ] ethyl } amino } carbonyl) -1H-pyrrol-3-yl ] -4-nitro-1H-pyrrol-2-amide (156mg, 0.33 mmol; see step (i) above) was suspended in methanol (25mL), to which was added Pd/C-10% (108mg) with stirring at 0 ℃ under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 5 hours. The catalyst was removed on celite and methanol was removed under reduced pressure to give the title compound, which was used without further purification.

Example 1

4- ({ [4- ({4- [ (E) -2- (3-methoxyphenyl) ethenyl)]Benzoyl } amino) -1-methyl-1H-pyrrol-2-yl]Carboxy } amino) -1-methyl-N- [2- (4-morpholinyl) ethyl]-1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added HBTU (100mg, 0.264mmol), 4- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoic acid (32mg, 0.124 mmol; see preparation 4 above) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (without work-up) to afford the title compound as a pale yellow solid with no apparent melting point (26mg, 29%).

¹H NMR(DMSO-d₆)：10.32(1H，s)，9.97(1H，s)，9.68(1H，br)，8.23(1H，t，5.6Hz)，7.97(2H，d，8.4Hz)，7.75(2H，d，J＝5.6Hz)，7.3-7.29(4H，m)，7.22(2H，m)，7.12(1H，d，J＝1.5Hz)，7.00(1H，d，J＝1.5Hz)，6.90(1H，dd，J＝1.5Hz & J＝3.7Hz)，3.99(2H，m)，3.88(3H，s)，3.83(3H，s)，3.81(3H，s)，3.67-3.55(6H，m)，3.27(2H，m)，3.14(2H，m)。

IR(KBr)：1681，1642，1577，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: the following are found: 611.2971 calculation of value C₃₄H₃₉N₆O₅ 611.2982。

Example 2

1-methyl-4- ({ [ 1-methyl-4- {4- [ (E) -2- (3-quinoline) ethenyl]Benzoyl } amino) -1H-pyrrole-2-]Carbonyl } amino) -N- [2- (4-morpholine) ethyl]-1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added HBTU (100mg, 0.264mmol), 4- [ (E) -2- (3-quinoline) ethenyl ] benzoic acid (34mg, 0.123 mmol; see preparation 5 above) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work-up required) to afford the title compound as a pale yellow solid with no apparent melting point (36mg, 39%).

¹H NMR(DMSO-d₆)：10.35(1H，s)，9.98(1H，s)，9.55(1H，br)，9.28(1H，d，J＝2.0Hz)，8.59(1H，d，J＝2.0Hz)，8.23(1H，t，J＝8.0Hz)，8.05-7.97(5H，m)，7.83-7.75(4H；m)，7.70-7.60(4H，m)，7.34(1H，d，J＝1.7Hz)，7.21(1H，d，J＝1.7Hz)，7.13(1H，d，J＝1.7Hz)，7.01(1H，d，J＝1.7Hz)，4.03-3.99(2H，m)，3.88(3H，s)，3.83(3H，s)，3.69-3.63(2H，m)，3.59-3.54(4H，m)，3.28(2H，m)，3.15(2H，m)。

IR(KBr)：1681，1642，1577，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: the following are found: 632.2982 calculation of value C₃₆H₃₈N₇O₄ 632.2985。

Example 3

1-methyl-N- [ 1-methyl-5- { [2- (4-morpholinyl) ethyl]Amino } carbonyl-1H-pyrrol-3-yl]-4- { [ (4- (E) -2- (1-methyl-1H-pyrrol-2-yl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

(i)1-methyl-4- { [ (1-methyl-4-amino-1H-pyrrol-2-yl) carbonyl]Amino } -N- [2- (4-morpholinyl) ethyl]-1H-pyrrole-2-amides

1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrole-2-carboxamide (86mg, 0.212 mmol; see preparation 1(iv) above) was suspended in methanol (25mL), to which was added Pd/C-10% (80mg) under nitrogen with stirring at 0 ℃. The reaction mixture was hydrogenated at room temperature under normal pressure for 4 hours. The catalyst was removed on celite and methanol was removed under reduced pressure to give the amine, which was dissolved in DMF (1mL, dry). The amine obtained was used in the next step without purification.

(ii)1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino) carbonyl } -1H-pyrrol-3-yl]-4- { [4- (E) -2- (1-methyl-1H-pyrrol-2-yl) ethenyl]Benzoyl amino-1H-pyrrole-2-amide, trifluoroacetate salt

4- (E) -2- (1-methyl-1H-pyrrol-2-yl) vinyl ] benzoic acid, lithium salt (39mg, 0.166 mmol; see preparation 6 above) was suspended in DMF (1mL, dried) to which was added HBTU (315mg, 0.830mmol) followed by the amine solution of step (i) above. The reaction mixture was left to stand at room temperature and stirred overnight. The product was purified by HPLC (no work up required) to give the title compound as a pale yellow solid (23mg, 16%) with no distinct melting point.

¹H NMR(DMSO-d₆)：10.26(1H，s)，9.96(1H，s)，9.55(1H，br)，8.22(1H，br)，7.93(2H，d，J＝8.4Hz)，7.68(2H，d，J＝8.4Hz)，7.32(2H，m)，7.20(1H，s)，7.11(1H，s)，7.00(1H，s)，6.95(1H，d，J＝16.2Hz)，6.82(1H，s)，6.53(1H，s)，6.06(1H，t，J＝3.1Hz)，4.02(2H，m)，3.87(3H，s)，3.83(3H，s)，3.72(3H，s)，3.69-3.66(2H，m)，3.63-3.53(4H，m)，3.26-3.23(2H，m)，3.14-3.11(2H，m)。

IR(KBr)：1681，1642，1577，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: the following are found: 584.2984 calculation of value C₃₂H₃₈N₇O₄ 584.2985。

Example 4

N- [5- ({ [3- (dimethylamino) propyl ] amino]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-4- ({4- [ (E) -2- (3-methoxyphenyl) ethenyl]Benzoyl } amino) -1-methyl-1H-pyrrole-2-amide, trifluoroacetate salt

To 4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1-methyl-1H-pyrrole-2-amide (41mg, 0.124 mmol; see preparation 2 above) in DMF (1mL) was added HBTU (141mg, 0.372mmol), 4- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoic acid (47mg, 0.186 mmol; see preparation 4 above) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work-up required) to give the title compound as a pale yellow solid (66mg, 51%) with no distinct melting point.

¹H NMR(DMSO-d₆)：10.32(1H，s)，9.94(1H，s)，9.38(1H，s)，8.15(1H，t，J＝5.8Hz)，7.98(2H，d，J＝8.4Hz)，7.75(1H，d，J＝8.4Hz)，7.37(2H，d，J＝1.5Hz)，7.33(2H，m)，7.21(2H，m)，7.19(1H, d, J ═ 1.5Hz), 7.11(1H, d, J ═ 1.5Hz), 6.95(1H, d, J ═ 1.5Hz), 8.89(1H, m), 3.87(3H, s), 3.82(3H, s), 3.81(3H, s), 3.25(2H, q, J ═ 6.1Hz), 3.06(2H, m), 2.79(6H, d, J ═ 4.8Hz), 1.84(2H, quintuple, J ═ 6.7 Hz).

IR(KBr)：1681，1642，1581，1540，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: found 583.3036 calculated value C₃₃H₃₉O₄N₆ 583.3033。

Example 5

N- [5- ({ [3- (dimethylamino) propyl ] amino]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

To 4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1-methyl-1H-pyrrol-2-amide (41mg, 0.124 mmol; see preparation 2 above) in DMF (1mL) was added HBTU (141mg, 0.372mmol), 4- [ (E) -2- (3-quinoline) vinyl ] benzoic acid (51mg, 0.186 mmol; see preparation 5 above) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to give the title compound as a pale yellow solid (15mg, 19%) with no distinct melting point.

¹H NMR(DMSO-d₆)：10.35(1H，s)，9.95(1H，s)，9.34(1H，br)，9.27(1H，d，J＝1.9Hz)，8.57(1H，d，J＝1.9Hz)，8.15(1H，t，J＝5.7Hz)，8.02(2H，d，J＝8.5Hz)，7.83(2H，d，J＝8.5Hz)，7.76(1H，dt，J＝1.4Hz &J6.9 Hz), 7.66(3H, m), 7.34(1H, d, J1.5 Hz), 7.19(1H, d, J1.5 Hz), 7.12(1H, d, J1.5 Hz), 6.95(1H, d, J1.5 Hz), 3.88(3H, s), 3.82(3H, s), 3.26(2H, q, J6.4 Hz), 3.10(2H, m), 2.79(6H, d, J4.9 Hz), 1.84(2H, quintuple, J7.9 Hz).

IR(KBr)：1681，1642，1577，1464，1436，1404，1266，1202，1134cm^-1。

HRFABMS: found 604.3038 calculated value C₃₅H₃₈O₃N₇ 604.3036。

Example 6

N- [5- ({ [3- (dimethylamino) propyl ] amino]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-4- ({3- [ (E) -2- (3-methoxyphenyl) ethenyl]Benzoyl) amino) -1-methyl-1H-pyrrole-2-amide, trifluoroacetate salt

To 4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1-methyl-1H-pyrrole-2-amide (41mg, 0.124 mmol; see preparation 2 above) in DMF (1mL) was added HBTU (141mg, 0.372mmol), 3- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoic acid (47mg, 0.186 mmol; see preparation 7 above) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to give the title compound as a pale yellow solid (67mg, 52%) with no distinct melting point.

¹H NMR(DMSO-d₆): 10.37(1H, s), 9.95(1H, s), 9.40(1H, br), 8.17(2H, m), 7.83(1H, d, J ═ 7.9Hz), 7.79(1H, d, j7.9hz), 7.52(1H, t, J ═ 7.7Hz), 7.36-7.29(4H, m), 7.22(2H, m), 7.19(1H, d, J ═ 1.5Hz), 6.95(1H, d, J ═ 1.5Hz), 6.89(1H, m), 3.88(3H, s), 3.82(3H, s), 3.81(3H, s), 3.25(2H, q, J ═ 6.0Hz), 3.06(2H, m), 2.79(6H, d, J ═ 6.84, J ═ 6.7H, 7.84 Hz), and quintuple (7H, 7.9Hz).

IR(KBr)：1682，1641，1578，1464，1436，1404，1266，1202，1134cm^-1。

HRFABMS: found 583.3029 calculated value C₃₃H₃₉O₄N₆583.3033。

Example 7

N- [5- ({ [3- (dimethylamino) propyl ] amino]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-1-methyl-4- ({4- [ (E) -2- (4-pyridinyl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

(i)1-methyl-4- { [ (1-methyl-4-amino-1H-pyrrol-2-yl) carbonyl]Amino } -N- [3- (dimethylamino) propyl group]-1H-pyrrole-2-amides

1-methyl 4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1H-pyrrole-2-amide (59mg, 0.156 mmol; see preparation 2(ii) above) was suspended in methanol (25mL), to which was added Pd/C-10% (59mg) under stirring at 0 ℃ under nitrogen. The reaction mixture was hydrogenated at room temperature under normal pressure for 3 hours. The catalyst was removed on celite and methanol was removed under reduced pressure to give the amine, which was dissolved in DMF (1.5mL, dry) and used without further purification.

(ii)N- [5- ({ [3- (dimethylamino) propyl ] amino]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-1-methyl-4- ({4- [ (E) -2- (4-pyridinyl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

HBTU (121mg, 0.32mmol), 4- [ (E)2- (4-Pyridyl radical) Vinyl radical]Benzoic acid (36mg, 0.16 mmol; see preparation 9 above) and NMM (0.30mL, dry). The reaction mixture was left to stir at room temperature overnight. The product was purified by HPLC (no work up required) to give the title compound as a pale yellow solid (33mg, 34%) with no distinct melting point.

¹H NMR(DMSO-d₆)：10.38(1H，s)，9.94(1H，s)，9.28(1H，br，TFA)，8.15(1H，t，J＝5.8Hz)，8.03(2H，d，J＝8.4Hz)，7.89(1H，d，J＝4.9Hz)，7.85(2H，d，J＝11.6Hz)，7.53(2H，d，J＝16.4Hz)，7.33(1H，d，J＝1.5Hz)，7.18(1H，d，J＝1.5Hz)，7.12(1H，d，J＝1.5Hz)，6.96(1H，d，J＝1.5Hz)，3.87(3H，s)，3.82(3H，s)，3.25(2H，q，J＝6.1Hz)，3.06(2H，m)，2.79(6H，d，J＝4.8Hz)，1.84(2H，Quintuple, J ═ 6.7 Hz).

IR(KBr)：1623，1638，1679，3000，3430cm^-1。

HRFABMS: the following are found: 554.2882 calculation of value C₃₁H₃₆N₇O₃ 554.2880。

Example 8

N- [5- ({ [5- ({ [3- (dimethylamino) propyl group)]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]Amino } carbonyl) -1-methyl-1H-pyrrol-3-yl]-1-methyl-4- [ (E) -2- (4-nitrophenyl) -vinyl]-1H-pyrrole-2-amide, trifluoroacetate salt

To 1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl ] -1H-pyrrole-2-carboxylic acid (40mg, 0.146 mmol; see preparation 8 above) and HBTU (111mg, 0.292mmol) was added a solution of 4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1-methyl-1H-pyrrole-2-amide (41mg, 0.146 mmol; see preparation 2 above) in DMF (1.5mL) with stirring at room temperature. The reaction mixture was left at room temperature overnight and then purified by HPLC without workup. Fractions containing the desired material were collected and lyophilized to give the title compound as an orange solid (15.1mg, 15%) with no apparent melting point.

¹H NMR(DMSO-d₆): 10.03(1H, s), 9.91(1H, s), 9.21(1H, br), 8.19(2H, d, J ═ 8.8Hz), 8.14(1H, t, J ═ 5.5Hz), 7.75(2H, d, J ═ 8.8Hz), 7.41(1H, d, J ═ 16.2Hz), 7.31(1H, d, J ═ 1.5Hz), 7.23(2H, d, J ═ 1.5Hz), 7.17(1H, d, J ═ 1.5Hz), 7.07(1H, d, J ═ 1.5Hz), 6.95(1H, d, J ═ 1.5Hz), 6.94(1H, d, J ═ 16.2), 3.90(3H, s), 3.86(3H, s), 3.82(3H, 5H, 3.06H, 3H, 5H, 3.79 (7.06 Hz), 7.7.7H, 7H, 7.06, 3H, 5Hz), 7.7.7.7.7.7.7H, 3H, 5Hz, 7.7.7.7H, 3H, 5Hz, 7H, 5Hz, 7.

IR(KBr)：1663，1551，1401，1288，1202，750cm^-1。

HRFABMS: the following are found: 601.2888 calculation of value C₃₁H₃₇N₈O₅ 601.2887。

Example 9

N- [3- (dimethylamino) propyl group]-5-isopentyl-2- (([ 1-methyl-4- ({4- (E) -2- (3-quinolyl) ethenyl)]Benzoyl } amino) -1H-pyrrol-2-yl]Carbonyl } amino) -1, 3-thiazole-4-carboxamide, trifluoroacetate salt

To 4- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -N- [3- (dimethylamino) propyl ] -1-methyl-1H-pyrrol-2-amide (49mg, 0.118 mmol; see preparation 2 above) in DMF (1mL) was added HBTU (90mg, 0.188mmol), 4- [ (E) -2- (3-quinolyl) vinyl ] benzoic acid (33mg, 0.188 mmol; see preparation 5) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to give the title compound as a yellow solid (21mg, 31%) with no distinct melting point.

¹H NMR(DMSO-d₆): 12.11(1H, s), 10.45(1H, s), 9.33(1H, br), 9.28(1H, d, J ═ 2.0Hz), 8.59(1H, d, J ═ 2.0Hz), 8.03(4H, m), 7.96(1H, t, J ═ 6.2Hz), 7.83(2H, d, J ═ 8.4Hz), 7.77(1H, t, J ═ 7.1Hz), 7.65(3H, m), 7.54(1H, d, J ═ 1.5Hz), 7.46(1H, d, J ═ 1.5Hz), 3.92(3H, s), 3.36(2H, q, J ═ 6.4Hz), 3.19(2H, t, J ═ 7.06, 3.7 (2H, 3.63, 2H, 8.84 Hz), 3.6H, J ═ 6.4Hz), 3.19(2H, t, J ═ 6.7.7, 3.4 Hz), 3.3.3.7H, 3.63 (3.4 Hz), 3.3.3.3.3.7H, 3.3.3.3.3H, 3.3, 3.7, 3H, 3.3H, 3H.

IR(KBr)：1681，1642，1581，1540，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: the following are found: 678.3228 calculation of value C₃₈H₄₄N₇O₃S 678.3226。

Example 10

1-methyl-N- [ 1-methyl-5- ({ [ (1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino) -carbonyl) -1H-pyrroles-3-yl]Amino) carbonyl) -1H-pyrrol-3-yl]-4- [ (E) -2- (3-quinolinyl) ethenyl]-1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-carboxamide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added HBTU (83mg, 0.22mmol), 1-methyl-4- [ (E) -2- (3-quinolinyl) ethenyl ] -1H-pyrrole-2-carboxylic acid (31mg, 0.11 mmol; see preparation 12 above) and 4-methylmorpholine (25. mu.L, 0.22mmol), the resulting solution was stirred for 16H before HPLC purification and the product fractions were lyophilized to give the title compound (0.023g, 26%).

m.p. ═ no apparent m.p.

v_max NaCl/cm^-1：3404υ(N-H)，3118υ(N-Me)，2926υ(Ar-H)，1676，1616υ(C＝O)，1553，1465υ(C＝C)，1257υ(C-N)1132υ(C-O-C)，720δ(C-H)。

δ_H ¹H(CDCl₃)：3.18(2H，m，CH₂)，3.38(2H，m，CH₂)，3.56(4H，m，CH₂)，3.83(3H，s，NMe)，3.87(3H，s，NMe)，3.91(3H，s，NMe)，4.00(2H，m，CH₂) 7.00(2H, m, Ar-H and (CH ═ CH)), 7.08(1H, d, Ar-H (J ═ 1.6Hz)), 7.21(1H, d, Ar-H (J ═ 1.6Hz)), 7.25(3H, m, Ar-H), 7.42(1H, d, (CH ═ CH) (J ═ 16.4Hz)), 7.62(1H, t, Ar-H (J ═ 7.2Hz)), 7.72(1H, t, Ar-H (J ═ 7.2Hz)), 7.98(2H, m, Ar-H), 8.25(1H, t, NH (J ═ 5.6Hz)), 8.43(1H, s, Ar-H), 9.16(1H, s, Ar-H), 9.86(1H, s) N (1H, s-H), and (N ═ 1H, s-H))⁺)，9.96(1H，s，NH)，10.04(1H，s，NH)。

HRFABMS: found 635.3083 calculated value C₃₅H₃₉N₈O₄ 635.3094。

Example 11

1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]-4-[({1-Methyl-4- [ (E) -2- (2-quinolyl) ethenyl]-1H-pyrrol-2-yl } carbonyl-amino]-1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-carboxamide (46mg, 0.124 mmol; see preparation 1 above) dissolved in DMF (1mL) was added HBTU (83mg, 0.22mmol), 1-methyl-4- [ (E) -2- (2-quinolinyl) ethenyl ] -1H-pyrrole-2-carboxylic acid (31mg, 0.11 mmol; see preparation 13 above) and 4-methylmorpholine (25. mu.L, 0.22mmol), the resulting solution was stirred for 16H before HPLC purification and the product fractions were lyophilized to give the title compound (0.027g, 26%).

m.p. ═ no apparent m.p.

v_max NaCl/cm^-1：3407υ(N-H)，3122υ(N-Me)，2924υ(Ar-H)，1678，1619υ(C＝O)，155，1464υ(C＝C)，1247υ(C-N)1131υ(C-O-C)，722δ(C-H)。

δ_H ¹H(CDCl₃)：3.17(2H，m，CH₂)，3.39(2H，m，CH₂)，3.57(4H，m，CH₂)，3.82(3H，s，NMe)，3.85(3H，s，NMe)，3.90(3H，s，NMe)，4.00(2H，m，CH₂) 7.00(1H, d, Ar-H (J ═ 1.6Hz)), 7.10(2H, m, Ar-H, and (H) C ═ CH), 7.21(1H, d, Ar-H (J ═ 1.6Hz)), 7.27(1H, d, Ar-H (J ═ 1.6Hz)), 7.34(1H, d, Ar-H (J ═ 1.6Hz)), 7.45(1H, d, Ar-H (J ═ 1.6Hz)), 7.62(2H, m, Ar-H), 7.83(2H, m, Ar-H), 8.02(3H, m, Ar-H), 8.25(1H, t, N-H (J ═ 5.6Hz)), 8.50(1H, s, NH, H, and NH ═ C), and C⁺)，9.89(1H，s NH⁺)，9.97(1H，s，NH)，10.12(1H，s，NH)。

LREIMS: found 635.27 calculated value C₃₅H₄₁N₈O₄ 635.31。

Example 12

N- [ 1-methyl-5- (([ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyridinePyrrol-3-yl]-2- [ (E) -2- (2-quinolinyl) ethenyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (50mg, 0.12 mmol; see preparation 1 above) dissolved in DMF (1mL) was added HBTU (83mg, 0.22mmol), 2- [ (E) -2- (2-quinolinyl) vinyl ] -1, 3-thiazole-4-carboxylic acid (35mg, 0.12 mmol; see preparation 14 above) and 4-methylmorpholine (25. mu.L, 0.22mmol), and the resulting solution was stirred for 16H before HPLC purification and the product fractions were lyophilized to give the title compound (0.020g, 21%).

m.p. ═ no apparent m.p.

v_max NaCl/cm^-1：3421υ(N-H)，3116υ(N-Me)，2928υ(Ar-H)，1677，1647，1638υ)(C＝O)，1556，1465υ(C＝C)，1241υ(C-N)1204，1131υ(C-O-C)，722δ(C-H)。

δ_H ¹H(CDCl₃)：3.17(2H，m，CH₂)，3.39(2H，m，CH₂)，3.57(4H，m，CH₂)，3.84(3H，s，NMe)，3.88(3H，s，NMe)，4.00(2H，m，CH₂)，7.00(1H，d，Ar-H(J＝1.6Hz))，7.22(2H，m，Ar-H)，7.35(1H，d，Ar-H(J＝1.6Hz))，7.63(1H，t，Ar-H(J＝6.9Hz))，7.80(1H，t，Ar-H(J＝6.9Hz))，7.90(1H，d，(H)C＝CH(J＝16.2Hz))，7.96(1H，d，Ar-H(J＝8.6Hz))，8.01(2H，m，Ar-H)，8.09(1H，d，(H)C＝CH(J＝16.2Hz))，8.24(1H，t N-H(J＝5.6Hz))，8.40(1H，s，Ar-H)，8.45(1H，d，Ar-H(J＝8.6Hz))，9.66(1H，s，NH⁺) 9.98(1H, s, N-H), 10.39(1H, s, N-H), calculated 634.30 for MS M/z M, found 635.27(M + H).

LREIMS: found 639.20 calculated value C₃₃H₃₅N₈O₄S 639.25。

Example 13

1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] ethaneBase of]Amino } carbonyl-1H-pyrrol-3-yl) -4- ({4- (E) -2- (2-naphthyl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added 4- [ (E) -2- (2-naphthyl) vinyl ] benzoic acid sodium salt (37mg, 0.124 mmol; see preparation 17 above) and HBTU (94mg, 0.248mmol) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) and after lyophilization gave the title compound as a pale yellow solid (20mg, 22%) with no apparent melting point.

¹H NMR(DMSO-d₆)：10.33(1H，s)，9.97(1H，s)，9.5(1H，br)，8.22(1H，t，J＝4.5Hz)，8.06(1H，s)，8.00-7.92(6H，m)，7.81(2H，d，J＝8.4Hz)，7.60-7.47(4H，m)，7.34(1H，d，J＝1.6Hz)，7.21(1H，d，J＝1.6Hz)，7.13(1H，d，J＝1.6Hz)，7.01(1H，d，J＝1.6Hz)，4.02(2H，m)，3.90(3H，s)，3.84(3H，s)，3.72-3.54(6，m)，3.28-3.12(4H，m)。

IR(KBr)：1681，1642，1581，1540，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: finding out; 631.3030 calculation of value C₃₇H₃₉N₆O₄ 631.3028。

Example 14

4- [ (4- { (E) -2- [2- (1H-1, 2, 3-benzotriazol-1-yloxy) -3-quinolinyl)]Vinyl benzoyl amino]-1-methyl-N- [ 1-methyl-5- { [2- (4-morpholinyl) ethyl]Amino } carbonyl-1H-pyrrol-3-yl]-1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added 4- [ (E) -2- (2-chloro-3-quinolinyl) ethenyl ] benzoic acid (38mg, 0.124 mmol; see preparation 16 above) and HBTU (94mg, 0.248mmol) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to afford the title compound as a lyophilized, light yellow solid (19mg, 17%) with no apparent melting point.

¹H NMR(DMSO-d₆)：10.39(1H，s)，9.97(1H，s)，9.55(1H，s)，9.03(1H，s)，8.24(2H，d，J＝8.3Hz)，8.08-8.02(3H，m)，7.93-7.89(2H，m)，7.85-7.77(3H，m)，7.66-7.54(4H，m)，7.37(1H，d，J＝7.8Hz)，7.34(1H，d，J＝1.6Hz)，7.21(1H，d，J＝1.6Hz)，7.14(1H，d，J＝1.6Hz)，7.01(1H，d，J＝1.6Hz)，4.02(2H，m)，3.88(3H，s)，3.86(3H，s)，3.69-3.53(6H，m)，3.27-3.14(4H，m)。

IR(KBr)：1681，1642，1581，1540，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: the following are found: 765.3267 calculation of value C₄₂H₄₁N₁₀O₅ 765.3261。

Example 15

1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]-4- ({4- [ (E) -2- (2-quinolyl) ethenyl]Benzoyl } amino) -1H-pyrrole-2-amide, trifluoroacetate salt

To 4-amino-1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrole-2-amide (46mg, 0.124 mmol; see preparation 1 above) in DMF (1mL) was added 4- [ (E) -2- (2-quinolinyl) ethenyl ] benzoic acid (34mg, 0.124 mmol; see preparation 15 above) and HBTU (94mg, 0.248mmol) with stirring at room temperature. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to afford the title compound as a lyophilized, light yellow solid (16mg, 20%) with no apparent melting point.

¹H NMR(DMSO-d₆): 10.39(1H, s), 10.08(1H, br), 9.99(1H, s), 8.42(1H, d, 8.7Hz), 8.27(1H, t, unresolved), 8.03-7.88(8H, m), 7.78(1H, t, J ═ 7.0Hz), 7.65-7.57(2H, m), 7.35(1H, d, J ═ 1.6H), 7.23(1H, d, J ═ 1.6Hz), 7.13(1H, s), 6.99(1H, d, J ═ 1.6Hz), 4.01(2H, m), 3.88(3H, s), 3.83(3H, s), 3.75-3.69(4H, m), 3.60-3.40(6H, m).

IR(KBr)：1681，1642，1581，1540，1464，1435，1404，1266，1202，1134cm^-1。

HRFABMS: found 632.2996 calculated value C₃₈N₇O₄ 632.2985。

Example 16

5-isopentyl-2- ({ [ 1-methyl-4- ((4- [ (E) -2- (3-quinolyl) ethenyl)]Benzoyl } amino) -1H-pyrrol-2-yl]Carbonyl } amino) -N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (75mg, 0.167 mmol; see preparation 18 above) dissolved in DMF (1mL) was added 4- [ (E) -2- (3-quinolyl) vinyl ] benzoic acid (46mg, 0.167 mmol; see preparation 5 above) and HBTU (126mg, 0.334mmol) at room temperature with stirring. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to afford the title compound as a lyophilized, light yellow solid (33mg, 24%) with no apparent melting point.

¹H NMR(DMSO-d₆)：12.09(1H，s)，10.46(1H，s)，9.71(1H，br)，9.28(1H，d，J＝1.6Hz)，8.58(1H，s)，8.10(1H，t，J＝5.8Hz)，8.03(4H，m)，7.83(2H，d，＝8.4Hz)，7.77(1H，dt，J＝8.2Hz & J＝1.4Hz)，7.71-7.60(3H，m)，7.54(1H，d，J＝1.6Hz)，7.45(1H，d，J＝1.6Hz)，4.02(2H，m)，3.93(3H，s)，3.70-3.54(6H，m)，3.31-3.15(6H，m)，1.64-1.51(3H，m)，0.93(6H，d，J＝6.3Hz)。

IR(KBr)：1671，1552，1288，1202，1134，834，799，721cm^-1。

HRFABMS: the following are found: 706.3179 calculation of value C₃₉H₄₄O₄N₇S 706.3176。

Example 17

2- { (4- ({4- [ (E) -2- (2-chloro-3-quinolinyl) ethenyl]Benzoyl } amino) -1-methyl-1H-pyrrol-2-yl]Carbonyl } amino) -5-isopentyl-N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (75mg, 0.167 mmol; see preparation 18 above) dissolved in DMF (1mL) was added 4- [ (E) -2- (2-naphthyl) vinyl ] benzoic acid sodium salt (46mg, 0.167 mmol; see preparation 16 above) and HBTU (126mg, 0.334mmol) at room temperature with stirring. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work-up required) to give the title compound as a lyophilized pale yellow solid (12mg, 9%) with no apparent melting point.

¹H NMR(DMSO-d₆)：12.08(1H，s)，10.49(1H，s)，9.72(1H，br)，9.03(1H，s)，8.24(1H，d，J＝8.4Hz)，8.08-7.99(3H，m)，7.94-7.88(1H，m)，7.85-7.79(2H，m)，7.66-7.56(3H，m)，7.54(1H，d，J＝1.6Hz)，7.46(1H，d，J＝1.6Hz)，7.37(1H，d，J＝7.9Hz)，4.02(2H，m)，3.93(3H，s)，3.70-3.54(6H，m)，3.31-3.15(6H，m)，1.64-1.51(3H，m)，0.93(6H，d，J＝6.3Hz)。

IR(KBr)：1663，1551，1502，1401，1288，1202，1137，778，750cm^-1。

HRFABMS: the following are found: 740.2786 calculation of value C₃₉H₄₃O₄N₇ ³⁵ClS 740.2737。

Example 18

2- [ { (4- (4- { (E) -2- [2- (1H-1, 2, 3-benzotriazol-1-yloxy) -3-quinolinyl)]Vinyl } -benzoyl) amino]-1-methyl-1H-pyrrol-2-yl } carbonyl) amino]-5-isopentyl-N- [2- (4-morpholinyl) ethyl-1, 3-thiazole-4-amide, trifluoroacetate salt

The reaction mixture described in example 17 above was isolated by HPLC purification to give the title compound as a yellow solid with no apparent melting point (19mg, 12%).

¹H NMR(DMSO-d₆)：12.09(1H，s)，10.49(1H，s)，9.68(1H，br)，9.03(1H，s)，8.93(1H，s)，8.23(1H，d，J＝8.4Hz)，8.09-7.95(6H，m)，7.90-7.77(6H，m)，7.70(1H，t，J＝7.0Hz)，7.65-7.57(4H，m)，7.55(1H，d，J＝1.6Hz)，7.46(1H，d，J＝1.6Hz)，7.37(1H，d，J＝9.1Hz)，12.08(1H，s)，10.49(1H，s)，9.72(1H，br)，9.03(1H，s)，8.24(1H，d，J＝8.4Hz)，8.08-7.99(3H，m)，7.94-7.88(1H，m)，7.85-7.79(2H，m)，7.66-7.56(3H，m)，7.54(1H，d，J＝1.6Hz)，7.46(1H，d，J＝1.6Hz)，7.37(1H，d，J＝7.9Hz)，4.02(2H，m)，3.93(3H，s)，3.70-3.54(6H，m)，3.31-3.15(6H，m)，1.64-1.51(3H，m)，0.93(6H，d，J＝6.3Hz)。

IR(KBr)：1663，1551，1502，1401，1288，1202，1137，778，750cm^-1。

HRFABMS: the following are found: 839.3455 calculation of value C₄₅H₄₇O₅N₁₀S 839.3452。

Example 19

5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) -2- (2-quinolyl) ethenyl)]Benzoyl } amino) -1H-pyrrol-2-yl]Carbonyl } amino) -N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (75mg, 0.167 mmol; see preparation 18 above) dissolved in DMF (1mL) was added 4- [ (E) -2- (2-chloro-3-quinolinyl) vinyl ] benzoic acid (46mg, 0.167 mmol; see preparation 15 above) and HBTU (126mg, 0.334mmol) at room temperature with stirring. The reaction mixture was left to stand at room temperature overnight. The title compound was purified by HPLC (no work up required) to afford the desired product as an orange solid after lyophilization (25mg, 18%) with no apparent melting point.

¹H NMR(DMSO-d₆)：12.09(1H，s)，10.49(1H，s)，9.82(1H，br)，8.45(1H，d，J＝8.6Hz)，8.11(1H，t，J＝5.7Hz)，8.05(2H，d，J＝8.3Hz)，8.00-7.93(4H，m)，7.92(2H，d，J＝8.4Hz)，7.79(1H，dd，J＝8.3 & J＝1.3Hz)，7.62(2H，m)，7.55(1H，d，J＝1.6Hz)，7.45(1H，d，J＝1.6Hz)，4.02(3H，m)，3.93(3H，s)，3.71(3H，m)，3.56(2H，m)，3.30(2H，m)，3.22-3.14(4H，m)，1.63-1.51(3H，m)，0.93(6H，d，J＝6.4Hz)。

IR(KBr)：1663，1551，1502，1401，1288，1202，1137，778，750cm^-1。

HRFABMS: the following are found: 706.3180 calculation of value C₃₉H₄₄O₄N₇S 706.3175。

Example 20

5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) -2- (2-naphthalenyl) ethenyl]Benzoyl } amino) -1H-pyrrol-2-yl]Carbonyl } amino) -N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (75mg, 0.167 mmol; see preparation 18 above) dissolved in DMF (1mL) was added 4- [ (E) -2- (2-naphthyl) vinyl ] benzoic acid sodium salt (50mg, 0.167 mmol; see preparation 17 above) and HBTU (126mg, 0.334mmol) at room temperature with stirring. The reaction mixture was left to stand at room temperature overnight. The title compound was purified by HPLC (no work up required) to afford the desired product as a light solid after lyophilization (19mg, 14%) with no apparent melting point.

¹H NMR(DMSO-d₆)：12.08(1H，s)，10.43(1H，s)，9.59(1H，br)，8.10-8.07(2H，m)，8.01(2H，d，J＝8.4Hz)，7.94-7.91(4H，m)，7.81(2H，d，J＝8.4Hz)，7.55-7.45(6H，m)，4.02(3H，m)，3.93(3H，s)，3.66(3H，m)，3.57(2H，m)，3.30(2H，m)，3.22-3.15(4H，m)，1.63-1.51(3H，m)，0.93(6H，d，J＝6.4Hz)。

IR(KBr)：1663，1551，1502，1401，1288，1202，1137，778，750cm^-1。

HRFABMS: the following are found: 705.3221 calculation of value C₄₀H₄₅O₄N₆S 705.3223。

Example 21

5-isopentyl-2- [ ({ 1-methyl-4- { [ (1-methyl-4- [ (E) -2- (4-morpholinyl) ethenyl)]-1H-pyrrol-2-yl } carbonyl) amino]-1H-pyrrol-2-yl } carbonyl) amino]-N- [2- (4-morpholinyl) ethyl]-1, 3-thiazole-4-amide, trifluoroacetate salt

To 2- { [ (4-amino-1-methyl-1H-pyrrol-2-yl) carbonyl ] amino } -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide (75mg, 0.167 mmol; see preparation 18 above) dissolved in DMF (1mL) was added 1-methyl-4- [ (E) -2- (4-naphthyl) vinyl ] -1H-pyrrole-2-carboxylic acid (46mg, 0.167 mmol; see preparation 8 above) and HBTU (126mg, 0.334mmol) at room temperature with stirring. The reaction mixture was left to stand at room temperature overnight. The product was purified by HPLC (no work up required) to afford the title compound as an orange solid after lyophilization (19mg, 14%) with no apparent melting point.

¹H NMR(DMSO-d₆)：12.05(1H，s)，10.15(1H，s)，9.78(1H，br)，8.20(2H，d，J＝8.8Hz)，8.09(1H，t，J＝5.7Hz)，7.76(2H，d，J＝8.8Hz)，7.46(1H，d，J＝1.6Hz)，7.41(1H，d，J＝16.3Hz)，7.39(1H，d，J＝1.6Hz)，7.32(1H，d，J＝1.6Hz)，7.27(1H，J＝1.6Hz)，6.95(1H，d，J＝16.3Hz)，4.02(3H，m)，3.90(6H，s)，3.66(3H，m)，3.57(2H，m)，3.30(2H，m)，3.22-3.15(4H，m)，1.63-1.51(3H，m)，0.93(6H，d，J＝6.4Hz)。

IR(KBr)：1663，1551，1502，1401，1288，1202，1137，778，750cm^-1。

HRFABMS: found 702.2950 calculated value C₃₅H₄₂O₆N₈S 702.2948。

Example 22

6- (E) -2- (4-methoxyphenyl) vinyl]-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl-1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]Nicotinamide, trifluoroacetate salt

(i)4-amino-1-methyl-N- [ 1-methyl-5- ({ [3- (4-morpholinyl) ethyl)]Amino } -carbonyl) -1H-pyrrol-3-yl]-1H-pyrrole-2-amides

1-methyl-4- { [ (1-methyl-4-nitro-1H-pyrrol-2-yl) carbonyl ] amino } -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrole-2-amide (200mg, 0.495 mmol; see preparation 1(iv) above) was dissolved in methanol (25mL) at 0 deg.C under nitrogen. Pd/C-10% (170mg) was added portionwise under nitrogen at 0 ℃. The reaction mixture was hydrogenated at room temperature under normal pressure for 4 hours. The catalyst was removed over celite and the solvent was removed under reduced pressure to give the sub-title compound. It was dissolved in DMF (1mL, dry) and divided into four equal parts which were used without further purification to prepare the compounds in examples 22 to 25.

(ii)6- [ (E) -2- (4-methoxyphenyl) vinyl]-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrole-3-]Nicotinamide, trifluoroacetate salt

(ii) to the first portion of DMF solution obtained in step (i) above, the following were added with stirring at room temperature: HBTU (94mg, 0.248mmol) and 6- [ (E) -2- (4-methoxyphenyl) vinyl ] nicotinic acid (32mg, 0.124 mmol; see preparation 21 above). The reaction mixture was left to stir at room temperature overnight, then the product was purified by HPLC to give the title compound as an orange solid (29mg, 32%) with no apparent melting point.

IR(KBr)：3427，1673，1588，1402，1253，1202，1174，832，720cm^-1。

¹H NMR(DMSO-d₆)：10.48(1H，s)，9.98(2H，s & br)，9.07(1H，s)，8.28(1H，d，J＝2.3Hz)，8.26(1H，d，J＝2.3Hz)，7.77(1H，d，J＝16.0Hz)，7.67(3H，d，J＝8.8Hz)，7.35(1H，d，J＝1.6Hz)，7.27(1H，d，J＝16.0Hz)，7.22(1H，d，J＝1.6Hz)，7.12(1H，d，J＝1.6Hz)，7.01(3H，m)，4.01(2H，m)，3.88(3H，s)，3.83(3H，s)，3.80(3H，s)，3.73(2H，m)，3.56(4H，m)，3.27(2H，m)，2.99(2H，m)。

HRFABMS: found 611.2990 calculated value C₃₄H₃₉O₅N₆ 611.2982。

Example 23

2- [ (E) -2- (4-methoxyphenyl) vinyl]-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-6-quinolinamides, trifluoroacetates

To the second portion of DMF solution obtained in step (i) of example 22 above, the following was added with stirring at room temperature: HBTU (94mg, 0.248mmol) and 2- [ (E) -2- (4-methoxyphenyl) vinyl ] -6-quinolinecarboxylic acid (38mg, 0.124 mmol; see preparation 22 above). The reaction mixture was left to stir at room temperature overnight, then the product was purified by HPLC to give the title compound as an orange solid (60mg, 63%) with no apparent melting point.

IR(KBr)：3422，1672，1583，1514，1249，1203，1173，1133，833，720cm^-1。

¹H NMR(DMSO-d₆)：10.58(1H，s)，10.00(1H，s)，9.84(1H，br)，8.55(1H，d，J＝1.6Hz)，8.53(1H，d，J＝8.7Hz)，8.28(2H，m)，8.09(1H，d，J＝8.8Hz)，7.98(1H，d，J＝8.7Hz)，7.94(1H，d，J＝16.2Hz)，7.73(2H，d，J＝8.6Hz)，7.41(1H，t，J＝4.6Hz)，7.38(1H，d，J＝1.6Hz)，7.23(1H，d，J＝1.6Hz)，7.16(1H，d，J＝1.6Hz)，7.04(3H，m)，4.02(2H，m)，3.89(3H，s)， 3.84(3H，s)，3.82(3H，s)，3.73(2H，m)，3.56(4H，m)，3.27(2H，m)，3.14(2H，m)。

HRFABMS: found 662.3089 calculated value C₃₇H₄₀O₅N₇ 662.3091。

Example 24

N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-2- ((E) -2- [4- (methylsulfanyl) -phenylvinyl } -6-quinolinamide, trifluoroacetate salt

To the third portion of DMF solution obtained in step (i) of example 22 above, the following was added with stirring at room temperature: HBTU (94mg, 0.248mmol) and 2- { (E) -2- [4- (methylthio) phenyl ] vinyl } -6-quinoline-carboxylic acid (40mg, 0.124 mmol; see preparation 20 above). The reaction mixture was left to stir at room temperature overnight, then the product was purified by HPLC to give the title compound as an orange solid (43mg, 43%) with no apparent melting point.

IR(KBr)：3424，1673，1581，1514，1250，1203，1173，1133，833，720cm^-1。

¹H NMR(DMSO-d₆)：¹H NMR(DMSO-d₆)：10.57(1H，s)，10.00(1H，s)，9.67(1H，br)，8.55(1H，d，J＝1.6Hz)，8.50(1H，d，J＝8.7Hz)，8.28(2H，m)，8.09(1H，d，J＝8.8Hz)，7.96(1H，d，J＝8.7Hz)，7.91(1H，d，J＝16.2Hz)，7.72(2H，d，J＝8.6Hz)，7.50(1H，t，J＝4.6Hz)，7.38(1H，d，J＝1.6Hz)，7.34(2H，d，J＝8.5Hz)，7.22(1H，d，J＝1.6Hz)，7.16(1H，d，J＝1.6Hz)，7.01(1H，d，J＝1.6Hz)，4.02(2H，m)，3.89(3H，s)，3.84(3H，s)，3.73(2H，m)，3.56(4H，m)，3.27(2H，m)，3.14(2H，m)，3.53(3H，s)。

HRFABMS: the following are found: 677.2788 calculation of value C₃₇H₃₉N₇O₄S 677.2784。

Example 25

N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl)]Amino } carbonyl) -1H-pyrrol-3-yl]Amino } carbonyl) -1H-pyrrol-3-yl]-6- { (E) -2- [4- (methylthio) -phenyl]Vinyl nicotinamide, trifluoroacetate salt

To the fourth portion of the DMF solution obtained in step (i) of example 22 above, the following was added with stirring at room temperature: HBTU (94mg, 0.248mmol) and 6- { (E) -2- [4- (methylthio) phenyl ] vinyl } nicotinic acid (34mg, 0.124 mmol; see preparation 19 above). The reaction mixture was left to stir at room temperature overnight, then the product was purified by HPLC to give the title compound as an orange solid (39mg, 42%) with no apparent melting point.

IR(KBr)：3424，1673，1581，1514，1250，1203，1173，1133，833，720 cm^-1。

¹H NMR(DMSO-d₆)：10.49(1H，s)，9.99(1H，s)，9.76(1H，br)，9.08(1H，s)，8.29-8.24(2H，m)，7.78(1H，d，J＝16Hz)，7.68(1H，d，J＝8.9Hz)，7.66(1H，d，J＝8.6Hz)，7.36-7.29(4H，m)，7.21(1H，d，J＝1.5Hz)，7.11(1H，d，J＝1.5Hz)，6.99(1H，d，J＝1.5Hz)，4.02(2H，m)，3.89(3H，s)，3.84(3H，s)，3.73(2H，m)，3.56(4H，m)，3.27(2H，m)，3.14(2H，m)，2.52(3H，s)。

HRFABMS: the following are found: 628.2710 calculation of value C₃₃H₃₈O₄N₇S 628.2706。

Example 26

The title compounds in the examples show bactericidal activity against a number of different microorganisms as detailed in table 1 below.

Organism

Bacteria

Staphylococcus aureus strain 1 (BSAC04(l)) (clinical from Glasgow)

Royal hospital separation)

Staphylococcus aureus strain 2 (NCTC 6571)

Streptococcus faecalis (NCTC 775)

Mycobacterium fortuitum (NCTC 10394)

Fungi

Candida albicans (NCPF3179)

Aspergillus niger (IMI 1745)

Table 1: the Minimum Inhibitory Concentrations (MICs) of the title compounds in the examples are indicated in. mu.M and (. mu.g/mL).

Keyword

not active at 100mg/mL (i.e., MIC > 100mg/mL)

not tested

Example 27

The title compounds of examples 2 and 22 showed bactericidal activity against a number of different strains of staphylococcus aureus as detailed in table 2 below. BSAC03/04/05(1) - (10) represent clinical strains of Staphylococcus aureus collected in clinical isolates in Scotland.

Table 2: the Minimum Inhibitory Concentrations (MICs) of the title compounds of examples 2 and 22 are expressed in μ g/mL.

Example 28

Determination of the minimum effective concentration of the title compound of example 2 in a staphylococcus aureus infection model.

Mice were infected with staphylococcus aureus LS-1 (5 million cfu i) and treated with a quantitative amount of the compound of example 2 (i.v. over 30 minutes). Each group included 5 mice and contained the following.

A first group: staphylococcus aureus LS-1 alone

Second group: staphylococcus aureus LS-1 plus 60mg/kg of the compound of example 2

Third group: staphylococcus aureus LS-1 plus 40mg/kg of the compound of example 2

And a fourth group: staphylococcus aureus LS-1 plus 20mg/kg of the compound of example 2

And a fifth group: staphylococcus aureus LS-1 plus 10mg/kg of the compound of example 2

Mice were assessed daily for weight loss, development of arthrocele and morbidity/mortality. The results are summarized in tables 3 to 5.

Table 3: development of arthrocele after infection

Table 4: mortality of mice after infection

Table 5: weight loss in mice after infection

Abbreviations:

br as broad (related to NMR)

CE-capillary electrophoresis

cfu is a colony-forming unit

d ═ bimodal (related to NMR)

DCM ═ dichloromethane

DMF ═ N, N-dimethylamide

DMSO ═ dimethyl sulfoxide

DNA-deoxyribonucleic acid

dsDNA ═ double-stranded deoxyribonucleic acid

eq. -. equivalent

FRET (fluorescence resonance energy transfer)

h is hour

HBTU ═ O-benzotriazol-1-yl-N, N' -tetramethyluronium hexafluorophosphate

HCl ═ hydrochloric acid

HOBT ═ 1-hydroxybenzotriazole

HPLC ═ high performance liquid chromatography

High resolution electron ionization mass spectrometry (HRIMS)

HRFABMS (high resolution fast atom bombardment Mass Spectrometry)

IR infrared (relating to spectrum)

iv is intravenous injection

LRESMS low resolution electrospray mass spectrometry

multiplet (related to NMR)

Me is methyl

min. (min.)

MIC is minimum inhibitory concentration

m.p. ═ melting point

MS mass spectrum

Vmax wave number (related to infrared spectrum)

NMM ═ N-methylmorpholine

NMR (nuclear magnetic resonance)

Pd/C-Palladium on carbon

q ═ quartet (related to NMR)

RT/RT ═ room temperature

single peak (related to NMR)

t-Tri-Peak (related to NMR)

TEA ═ triethylamine

THF ═ tetrahydrofuran

TFA-trifluoroacetic acid

The prefixes n-, s-, i-, t-and tert-have the usual meanings: normal, secondary, iso and tertiary.

Claims (19)

1. A compound of formula I or a pharmaceutically acceptable salt thereof:

wherein

Wavy lines indicate trans stereochemistry;

R¹-Q^arepresents methoxyphenyl, quinolyl, 1-methyl-2-pyrrolyl, pyridyl, naphthyl, 1, 2, 3-benzotriazoleOxyquinolinyl, methylthiophenyl, nitrophenyl or chloroquinolinyl;

Q^brepresents phenyl, pyridyl, quinolyl or a structural fragment of Ia or Id, wherein G¹Represents CH, R⁴Represents H, R⁵Represents methyl, and R⁸Represents H;

Q^crepresents a structural fragment of Ia or Id, wherein G¹Represents CH, R⁴Represents H, R⁵Represents methyl, and R⁸Represents C_1-12An alkyl group;

a represents 2;

a represents C_2-6An alkylene group; and

d represents 4-morpholinyl, -N (CH)₃)₂，

With the proviso that when R¹-Q^aWhen it represents 1-methyl-2-pyrrolyl, Q^bA structural fragment other than Ia; and when Q^bWhen represents a structural fragment of Ia, R¹-Q^aIs not 1-methyl-2-pyrrolyl.

2. The compound of claim 1, which is a compound of general formula Ig:

wherein

Wavy lines indicate trans stereochemistry;

R¹-Q^arepresents methoxyphenyl, quinolyl, 1-methyl-2-pyrrolyl, pyridyl, naphthyl, 1, 2, 3-benzotriazolyloxyquinolyl, methylthiophenyl, nitrophenyl or chloroquinolyl;

Q^crepresents a structural fragment of Ia or Id, wherein G¹Represents CH, R⁴Represents H, R⁵Represents methyl, and R⁸RepresentsC_1-12An alkyl group;

a represents 2;

G²represents CH or N;

a represents C_2-6An alkylene group; and

d represents 4-morpholinyl, -N (CH)₃)₂。

3. The compound of claim 1, wherein the compound is:

(i)4- ({ [4- ({4- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoyl } amino) -1-methyl-1H-pyrrol-2-yl ] carbonyl } amino) -1-methyl-N- [2- (4-morpholinyl) ethyl ] -1H-pyrrole-2-amide;

(ii) 1-methyl-4- ({ [ 1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-yl ] carbonyl } amino) -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrole-2-amide;

(iii) 1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -4- ({4- [ (E) -2- (1-methyl-1H-pyrrol-2-yl) ethenyl ] benzoyl } amino) -1H-pyrrole-2-amide;

(iv) n- [5- ({ [3- (dimethylamino) propyl ] amino } carbonyl) -1-methyl-1H-pyrrol-3-yl ] -4- ({4- [ E) -2- (3-methoxyphenyl) ethenyl ] benzoyl } amino) -1-methyl-1H-pyrrole-2-amide;

(v) n- [5- ({ [3- (dimethylamino) propyl ] amino } carbonyl) -1-methyl-1H-pyrrol-3-yl ] -1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl ] benzoyl } amino) -1H-pyrrole-2-amide;

(vi) n- [5- ({ [3- (dimethylamino) propyl ] amino } carbonyl) -1-methyl-1H-pyrrol-3-yl ] -4- ({3- [ (E) -2- (3-methoxyphenyl) vinyl ] benzoyl } amino) -1-methyl-1H-pyrrole-2-amide;

(vii) n- [5- ({ [3- (dimethylamino) propyl ] amino } carbonyl) -1-methyl-1H-pyrrol-3-yl ] -1-methyl-4- ({4- [ (E) -2- (4-pyridyl) ethenyl ] benzoyl } amino) -1H-pyrrole-2-amide;

(viii) n- [5- ({ [5- ({ [3- (dimethylamino) propyl ] amino } carbonyl) -1-methyl-1H-pyrrol-3-yl ] -1-methyl-4- [ (E) -2- (4-nitrophenyl) ethenyl ] -1H-pyrrol-2-amide;

(ix) n- [3- (dimethylamino) propyl ] -5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-yl ] carbonyl } amino) -1, 3-thiazole-4-amide;

(x) 1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] amino } carbonyl) -1H-pyrrol-3-yl ] -4- [ (E) -2- (3-quinolinyl) ethenyl ] -1H-pyrrol-2-amide;

(xi) 1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -4- [ ({ 1-methyl-4- [ (E) -2- (2-quinolinyl) ethenyl ] -1H-pyrrol-2-yl } carbonyl) amino ] -1H-pyrrol-2-amide;

(xii) N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -2- [ (E) -2- (2-quinolinyl) ethenyl ] -1, 3-thiazole-4-amide;

(xiii) 1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -4- ({4- [ (E) -2- (2-naphthyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-amide;

(xiv)4- [ (4- { (E) -2- [2- (1H-1, 2, 3-benzotriazol-1-yloxy) -3-quinolinyl ] vinyl } -benzoyl) amino ] -1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } -carbonyl) -1H-pyrrol-3-yl ] -1H-pyrrol-2-amide;

(xv) 1-methyl-N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -4- ({4- [ (E) -2- (2-quinolinyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-amide;

(xvi) 5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl ] benzoyl } -amino) -1H-pyrrol-2-yl ] carbonyl } amino) -N' - [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xvii)2- ({ [4- ({4- [ (E) -2- (2-chloro-3-quinolinyl) ethenyl ] benzoyl } amino) -1-methyl-1H-pyrrol-2-yl ] carbonyl } amino) -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xviii)2- [ ({4- [ (4- { (E) -2- [2- (1H-1, 2, 3-benzotriazol-1-yloxy) -3-quinolinyl ] -vinyl } benzoyl) amino ] -1-methyl-1H-pyrrol-2-yl } carbonyl) amino ] -5-isopentyl-N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xix) 5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) - (2- (2-quinolinyl) ethenyl ] benzoyl } -amino) -1H-pyrrol-2-yl ] carbonyl } amino) -N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xx) 5-isopentyl-2- ({ [ 1-methyl-4- ({4- [ (E) -2- (2-naphthyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-yl ] carbonyl } amino) -N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xxi) 5-isopentyl-2- [ ({ 1-methyl-4- [ (E) -2- (4-nitrophenyl) vinyl ] -1H-pyrrol-2-yl } carbonyl) amino ] -N- [2- (4-morpholinyl) ethyl ] -1, 3-thiazole-4-amide;

(xxii)6- [ (E) -2- (4-methoxyphenyl) vinyl ] -N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] nicotinamide;

(xxiii)2- [ (E) -2- (4-methoxyphenyl) vinyl ] -N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -6-quinolinecarboxamide;

(xxiv) N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -2- { (E) -2- [4- (methylsulfanyl) -phenyl ] vinyl } -6-quinolinecarboxamide;

(xxv) N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] -6- { (E) -2- [4- (methylsulfanyl) -phenyl ] ethenyl } nicotinamide,

or a pharmaceutically acceptable salt thereof.

4. The compound of claim 1, wherein the compound is 4- ({ [4- ({4- [ (E) -2- (3-methoxyphenyl) ethenyl ] benzoyl } amino) -1-methyl-1H-pyrrol-2-yl ] carbonyl } amino) -1-methyl-N- [2- (4-morpholinyl) ethyl ] -1H-pyrrol-2-amide or a pharmaceutically acceptable salt thereof.

5. The compound of claim 1, wherein the compound is 1-methyl-4- ({ [ 1-methyl-4- ({4- [ (E) -2- (3-quinolinyl) ethenyl ] benzoyl } amino) -1H-pyrrol-2-yl ] carbonyl } amino) -N- [2- (4-morpholinyl) ethyl ] -1H-pyrrol-2-carboxamide or a pharmaceutically acceptable salt thereof.

6. The compound of claim 1, wherein the compound is 6- [ (E) -2- (4-methoxyphenyl) ethenyl ] -N- [ 1-methyl-5- ({ [2- (4-morpholinyl) ethyl ] amino } carbonyl) -1H-pyrrol-3-yl ] nicotinamide or a pharmaceutically acceptable salt thereof.

7. A formulation including a compound as claimed in claim 1 in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

8. A formulation for use in the treatment of a bacterial, fungal or parasitic infection, which comprises a compound as claimed in claim 1 in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

9. The formulation of claim 8, wherein the disease is malaria.

10. A compound according to claim 1 for use as a medicament.

11. A compound according to claim 1 for use as a medicament for the treatment of a bacterial, fungal or parasitic infection.

12. A compound according to claim 11 wherein the disease is malaria.

13. Use of a compound according to claim 1 as active ingredient in the manufacture of a medicament for the treatment of a bacterial, fungal or parasitic infection.

14. The use according to claim 13, wherein the disease is malaria.

15. A combination product comprising the following components:

(A) a composition comprising the compound of claim 1, and

(B) compositions comprising one or more chemical agents known to be effective in treating diseases that depend on DNA replication for transmission.

16. A combination product according to claim 15 wherein each of (a) and (B) is formulated in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

17. A combination kit comprising the following components:

(A) a composition comprising the compound of claim 1, and

(B) compositions comprising one or more chemical agents known to be effective in treating diseases that are dependent on DNA replication,

wherein (A) and (B) are present as separate components.

18. The combination product of claim 15 or 16, wherein (a) and (B) are present as a single composition.

19. A process for the preparation of a compound of formula I according to claim 1, which process comprises:

(a) reaction of a compound of formula III with a compound of formula IV:

wherein Q^cD and A are as defined in claim 1 and c is as defined below,

wherein L is¹Represents a leaving group, c and d both represent integers from 0 to 4, wherein the sum of c and d is 2 and R is¹And Q^aTo Q^cAs defined in claim 1;

(b) reaction of a compound of formula Va with a compound of formula VI:

wherein R is¹⁵Represents aryl, the latter radical being selected from halogen, cyano, nitro, C_1-4Alkyl and C_1-4One or more substituents of alkoxy being optionally substituted, or R¹⁵Represents C_1-6Alkyl, and R¹And Q^aAs defined in claim 1, wherein the first and second substrates are,

wherein A, a, D, Q^bAnd Q^cAs defined in claim 1;

(c) reaction of a compound of formula Vb with a suitable base followed by reaction with a compound of formula VI as defined above:

wherein R is¹⁶Represents C_1-6Alkyl, and R¹And Q^aAs defined in claim 1, wherein the first and second substrates are,

(d) reacting a compound of formula VIIa with a compound of formula VIII:

wherein Q^b、Q^cA, A and D are as defined in claim 1 and R¹⁵As defined above, the above-mentioned,

wherein R is¹And Q^aAs defined in claim 1; or

(e) Reaction of a compound of formula VIIb with a suitable base, followed by reaction with a compound of formula VIII as defined above:

wherein Q^b、Q^cA, A and D are as defined in claim 1 and R¹⁶As defined above in (c).