Patent Description:
In <CIT> we described tri-and tetra-substituted naphthalene diimides and processes for producing them. None of the exemplified products that were tri-substituted had different amino-functional ligands at equational and polar positions on the core ligand. The method said to be suited for producing tri-substituted compounds was based on the following schematic:
<CHM>
where R<NUM> is optionally substituted alkyl or aryl and n is <NUM> or <NUM>. In practice a mixture of the tetra- (n=<NUM>) and tri-substituted (n=<NUM>) compounds was produced. All substituents, i.e. R<NUM> groups, are the same.

The specification describes methods for producing tetra-substituted compounds starting from the dichlorosubstituted analogue of the dibromo compound used above. The processes proceeded in one step, in which case the same H<NUM>NR<NUM> reagent reacted at both anhydride groups and both -chlorine-substituted carbon to give <NUM> identical R<NUM> substituents on the product, or in two steps where in the first step a first reagent H<NUM>NR<NUM> is reacted at both the anhydride groups and in a second step a second reagent H<NUM>NR<NUM> is reacted at both chlorine-substituted carbon atoms. Compounds with basic substituents on the imide substituent and/or on the aromatic rings have strong DNA quadruplex binding properties.

In <CIT> we described tri-substituted naphthalene diimides and processes for producing them. The method said to be suited for producing tri-substituted compounds was based on the following schematic:
<CHM>
where Y is H or Br, the group R<NUM> are the same and are selected from the group consisting of straight and branched chain C<NUM>-<NUM> alkenediyl, R<NUM> is selected from the group consisting of H and C<NUM>-<NUM> alkyl, R<NUM> is selected from the group consisting of straight and branched chain C<NUM>-<NUM> alkanediyl and C<NUM>-<NUM> aralkanediyl, X<NUM> is selected from the group consisting of halo, R<NUM>, NR<NUM><NUM>, CONR<NUM><NUM>, COOR<NUM>, SH and COR<NUM>, R<NUM> is selected from the group consisting of H, optionally substituted C<NUM>-<NUM> alkyl, optionally substituted C<NUM>-<NUM> cycloalkyl, C<NUM>-<NUM> heterocycloalkyl and aryl, each R<NUM> is selected from the group consisting of H, C<NUM>-<NUM> alkyl, aryl and C<NUM>-<NUM> aralkyl, N the groups R<NUM> together with the N-atom to which they are attached form a saturated heterocyclic ring of <NUM>-<NUM> atoms, each R<NUM> is selected from the group consisting of H and C<NUM>-<NUM> alkyl groups or the groups R<NUM> together with the N atom to which they are attached form a <NUM>-<NUM> membered heterocyclic ring, R<NUM> is selected from the group consisting of optionally substituted C<NUM>-<NUM> alkyl, C<NUM>-<NUM> aralkyl and aryl, R<NUM> is selected from the group consisting of optionally substituted C<NUM>-<NUM> alkyl, C<NUM>-<NUM> aralkyl and aryl, and whereby the Br atom or one of the or each Br atom is substituted by the nucleophilic amine nitrogen of the amine reagent to form the substituted NDI compound.

The tetra-substituted products, including products with groups R<NUM> different to groups R<NUM>, have been tested in <CIT>, <CIT> and in <NPL>, <NPL>, <NPL>, <NPL>, <NPL> and <NPL> for their binding properties to quadruplexes of telomeres and also those found in the promoter region of some genes. The data show the effective down-regulation of several proteins, the promoters of whose genes are targeted by the diimides, and hence result in growth inhibition of several cell-lines from a panel of cancer cell-lines. We have proposed in these publications to investigate further the impact of changing the nature of the substituent groups and the basicity of the tertiary amine groups in the cationic-substituents, on binding specificity and strength, and to investigate the potential of the compounds in cancer treatment, by testing models of cancers including pancreatic cancer.

In <NPL>, et al. , disclose the activity of <NUM>,<NUM>-bis((<NUM>-(<NUM>-methylpiperazine-l-yl)-propyl)amino)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone, also known as M M41 , in vivo in a mouse model of human pancreatic cancer.

<NPL>, disclose a tri-substituted naphthalene diimide compound, having <NUM>-dimethylamino ethyl groups substituted at each imido nitrogen atom and having, as the third substituent a <NUM>-(<NUM>-hydroxy-<NUM>-dimethyl amino methyl phenyl)ethyl amino group substituted at the <NUM>-position on the NDI core. It has activity stabilising the telomeric G-quadruplex (GQ), causing telomere dysfunction and telomerase down regulation. Global gene expression on a panel of cell lines showed modulation of genes implicated in telomere function and mechanisms of cancer. However the authors conclude that direct evidence for the biological relevance of G-<NUM> in the cell context is still lacking (<NPL>).

The synthesis of the tri-substituted compound reported by Nadai et al. is disclosed in <NPL>.

It has surprisingly been found by the inventors that a particular group of side-chains on a tetra-substituted naphthalene diimide compound results in improved binding of the diimide compound to GQ resulting in improved anti-cancer activity.

Accordingly, in a first aspect of the invention there is provided a new compound of Formula I as defined in claim <NUM>.

The invention further provides the new compounds for use in a method of treatment of an animal to treat cancer or to inhibit the growth of a solid tumour, or to reduce the size of a solid tumour, for instance pancreatic and prostate tumours.

The invention also provides compositions containing the new compound and a diluent or carrier. The compositions are preferably pharmaceutical compositions and the carrier is then pharmaceutically acceptable.

In a second aspect of the invention there is provided a method for synthesising a substituted naphthalene diimide compound according to the first aspect of the invention, comprising the steps of:.

wherein L, X and R<NUM> to R<NUM> are as defined for Formula I of the first aspect of the invention.

<FIG> shows the tumour regression in a pancreatic cancer tumour in mice treated with a compound of the invention and comparative compounds.

As used herein, "alkyl", "cycloalkyl", "heterocycloalkyl", "heterocyclic", "aryl", and "aralkyl" groups may be monovalent or divalent unless otherwise specified.

As used herein, unless otherwise specified "aryl" means a monocyclic, bicyclic, or tricyclic monovalent or divalent (as appropriate) aromatic radical, such as phenyl, biphenyl, naphthyl, anthracenyl, which can be optionally substituted with up to three substituent.

As used herein, unless otherwise specified "optionally substituted" is with an of the substituents selected from the group of C<NUM>-C<NUM> alkyl, hydroxy, C<NUM>-C<NUM> hydroxyalkyl, C<NUM>-C<NUM> alkoxy, C<NUM>-C<NUM> haloalkoxy, amino, C<NUM>-C<NUM> mono alkylamino, C<NUM>-C<NUM> bis alkylamino, C<NUM>-C<NUM> acylamino, C<NUM>-C<NUM> aminoalkyl, mono (C<NUM>-C<NUM> alkyl) amino C<NUM>-C<NUM> alkyl, bis(C<NUM>-C<NUM> alkyl) amino C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM>-acylamino, C<NUM>-C<NUM> alkyl sulfonylamino, halo, nitro, cyano, trifluoromethyl, carboxy, C<NUM>-C<NUM> alkoxycarbonyl, aminocarbonyl, mono C<NUM>-C<NUM> alkyl aminocarbonyl, bis C<NUM>-C<NUM> alkyl aminocarbonyl, -SO<NUM>H, C<NUM>-C<NUM> alkylsulfonyl, aminosulfonyl, mono C<NUM>-C<NUM> alkyl aminosulfonyl and bis C<NUM>-C<NUM>-alkyl aminosulfonyl.

As used herein, unless otherwise specified "heterocycloalkyl" and "heterocyclic" groups are carbocyclic radicals containing up to <NUM> heteroatoms selected from oxygen, nitrogen and sulfur. They may be bicyclic or monocyclic. They are preferably saturated. If the heterocycle is a divalent linker, the heterocycle may be attached to neighbouring groups through a carbon atom, or through one of the heteroatoms, e.g. a nitrogen atom. Examples of heterocycles are pyrrolidine, piperazine, and morpholine.

In the first aspect of the invention, L is preferably (CH<NUM>)<NUM>-<NUM>, even more preferably (CH<NUM>). Preferably L is in the para position of the phenyl. When R<NUM> is the optionally substituted nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl or the NR<NUM>R<NUM>, it is preferable that R<NUM> is joined to L via the nitrogen atom of R<NUM>.

It is envisaged that L or R<NUM> comprises a basic nitrogen atom. R<NUM> is preferably a nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl, preferably a nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl, more preferably a nitrogen-containing <NUM> membered heterocycloalkyl. Preferably the nitrogen of the nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl is the only heteroatom in the heterocycloalkyl. In another aspect, the nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl comprises a second heteroatom, such as an oxygen atom.

Suitably the nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl is selected from the group consisting of pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl, diazepanyl, preferably pyrrolidinyl. Suitably L is (CH<NUM>) and R<NUM> is pyrrolidinyl.

Suitably R<NUM> is NR<NUM>R<NUM>. R<NUM> and R<NUM> are independently C<NUM>-<NUM> alkyl, more preferably C<NUM>-<NUM> alkyl, even more preferably C<NUM>-<NUM> alkyl. Suitably the NR<NUM>R<NUM> is diethylamino, dipropylamino or ethylpropylamino.

In another embodiment, L comprises the basic nitrogen atom. Suitably L is (CH<NUM>)<NUM>-<NUM>NH, preferably (CH<NUM>)<NUM>-<NUM>NH, more preferably (CH<NUM>)<NUM>-<NUM>NH, even more preferably (CH<NUM>)NH and R<NUM> is a C<NUM>-<NUM>cycloalkyl, preferably a Cscycloalkyl. Preferably L is in the para position of the phenyl.

Both R<NUM> groups in Formula I are the same as one another. R<NUM> is a straight chain C<NUM>-<NUM>-alkanediyl, most preferably straight chain C<NUM>-alkanediyl. R<NUM> may or may not be the same as R<NUM>, and is a straight chain C<NUM>-<NUM>-alkanediyl, most preferably C<NUM>-alkanediyl.

X is NR<NUM><NUM>. The R<NUM> groups together with the N-atom to which they are attached form pyrrolidin-<NUM>-yl.

Preferably, Formula I has the following structure of Formula II:
<CHM>
wherein L and R<NUM> are as defined for Formula I, with any of the preferred groups as outlined above.

Suitably the compound is selected from the group consisting of: <NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)-<NUM>-(<NUM>-(pyrrolidin-<NUM>-ylmethyl)phenyl)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>-(<NUM>-(morpholinomethyl)phenyl)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)-<NUM>-(<NUM>-(pyrrolidin-<NUM>-ylmethyl)phenyl)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-(<NUM>-(piperidin-<NUM>-ylmethyl)phenyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>-(<NUM>-((diethylamino)methyl)phenyl)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>-(<NUM>-((cyclopentylamino)methyl)phenyl)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>-(<NUM>-(azepan-<NUM>-ylmethyl)phenyl)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; <NUM>-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methyl)phenyl)-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone; and
salts, hydrates and solvates thereof.

In the second aspect of the invention, L, X and R<NUM> to R<NUM> for Formula III to Formula VI are preferably as defined as the preferred features above for L, X and R<NUM> to R<NUM> of Formula I of the first aspect of the invention.

The method of the invention comprises a first step of reacting a brominated naphthalene diimide of Formula III with an amine reagent of Formula IV in an aromatic nucleophilic substitution reaction whereby the bromine atom is replaced by an amino group N(R<NUM>)(R<NUM>X). The starting diimide is a dibromo compound, and the aromatic nucleophilic substitution reaction may result in both bromine atoms being replaced by an amine group or just one of them (i.e. a compound of Formula V), although it is preferred that just one of the bromines is replaced, and there must be at least one compound of Formula V produced. It is preferable to separate a mixture of both the singly and doubly substituted naphthalene diimide, for example by using column chromatography.

In a second step, the compound of Formula V produced in the first step is reacted with a reagent of Formula VI in a substitution reaction whereby the bromine atom is replaced by the phenyl in Formula VI via the carbon atom that the LG (leaving group) is attached to initially. In one aspect the LG may be a boronic acid group, however, the skilled person will appreciate there are multiple ways to undergo the substitution reaction and form the aryl-aryl bond between Formulas V and VI. As a result, at least one compound of Formula I is produced.

Preferably, in another step, the compound of Formula I is isolated by using column chromatography.

Preferably, the specific form of column chromatography used is selected from gel and flash column chromatography.

The compounds of the present invention may be provided in the form of pharmaceutically acceptable compositions. The compounds of the present invention, especially when presented in the form of acid addition salts, for instance where some or all of the basic amine groups are converted to salt form, are water soluble and have approximately neutral pH. As such these salts are suitable for administration in the form of aqueous solution, which would be appropriate for intravenous administration. The pharmaceutical aqueous solutions preferably comprise <NUM> to <NUM>/I, of the compound.

The compounds of the present invention may be provided in a form suitable to be made up into pharmaceutical compositions, for instance, in dried, rehydratable form, for instance with carrier or diluent. Such dried forms may be produced by crystallisation and/or evaporation. Alternatively, the compounds may be presented as concentrates, for instance in water or an organic, pharmaceutically acceptable, solvent for dilution before administration.

As used herein, a pharmaceutically acceptable salt is a salt with a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include both inorganic acids such as hydrochloric, sulphuric, phosphoric, diphosphoric, hydrobromic or nitric acid and organic acids such as citric, fumaric, maleic, malic, ascorbic, succinic, tartaric, benzoic, acetic, methanesulfonic, ethanesulfonic, salicylic, stearic, benzenesulfonic or p-toluenesulfonic acid. Pharmaceutically acceptable bases include alkali metal (e.g. sodium or potassium) and alkali earth metal (e.g. calcium or magnesium) hydroxides and organic bases such as alkyl amines, aryl amines or heterocyclic amines.

Combinations according to the present invention may also be used in conjunction with other agents to inhibit undesirable and uncontrolled cell proliferation, for example antibodies. The compound may be conjugated to the antibody or administered as two separate components.

The compounds of the invention and compositions comprising them may be administered by any route. In one embodiment, a pharmaceutical composition comprising a compound of the invention may be formulated in a format suitable for oral, rectal, parenteral, intranasal or transdermal administration or administration by inhalation or by suppository. Typical routes of administration are parenteral, intranasal or transdermal administration or administration by inhalation. For chemotherapy of tumours, the compositions are most conveniently administered intravenously.

When used as treatment for existing tumours, the compounds of the present invention may be administered using regimens developed for chemotherapeutic agents.

The compounds of the invention and compositions have utility in treating subjects who have cancer. One particular class of cancers are known as solid tumours, in which a solid mass of cancerous material can be identified. Another class comprises haematological cancers, known as cancers that affect the blood system.

Specific types of cancers that can be treated using the compounds and compositions of the present invention include, but are not limited to prostate, pancreatic, small cell lung or gastro-intestinal. In a preferred embodiment, the cancer is prostate or pancreatic.

The compounds of the invention and compositions are useful in treatment to inhibit the growth of a solid tumour, or to reduce the size of a solid tumour, for example wherein the tumour is a pancreatic or prostate tumour.

The subject to be treated is suitably an animal, preferably a human.

The invention is further illustrated in the accompanying examples.

A series of tetrasubstituted naphthalene diimides have been synthesised and evaluated as G-quadruplex ligands, and as potential anti-cancer agents.

All chemicals, reagents, and solvents were purchased from commercial sources and used as received unless otherwise stated. Solvents were commercial HPLC grade unless dry solvent is specified, in which case the Aldrich 'Sure Seal' dry solvents were used. Column chromatography was performed on pre-packed silica (<NUM>-<NUM> mesh, <NUM>-<NUM>) cartridges using the eluent indicated.

<NUM>H NMR Spectra were acquired on a Bruker Avance III spectrometer at <NUM> using residual undeuterated solvent as reference.

Analytical LCMS was carried out using either acidic or basic methods as follows:.

Alternatively analytical UPLC/MS was carried out using either acidic or basic methods as follows:.

Preparative HPLC was carried out using a Waters Xselect CSH C18, <NUM>, 19x50 mm column using either a gradient of either <NUM>% Formic Acid in MeCN in <NUM>% aqueous Formic Acid or a gradient of MeCN in aqueous <NUM> Ammonium Bicarbonate; or a Waters Xbridge BEH C18, <NUM>, 19x50 mm column using a gradient MeCN in aqueous <NUM> Ammonium Bicarbonate. Fractions were collected following detection by UV at a single wavelength measured by a variable wavelength detector on a Gilson <NUM> preparative HPLC or Varian PrepStar preparative HPLC; by mass and UV at a single wavelength measured by a ZQ single quadrupole mass spectrometer, with positive and negative ion electrospray, and a dual wavelength detector on a Waters FractionLynx LCMS.

<NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthr oline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol), (<NUM>,<NUM>-dimethoxyphenyl)boronic acid (<NUM>, <NUM> mmol) or <NUM>-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)pyrrolidine (<NUM>, <NUM> mmol) and Pd(Ph3P)<NUM> (<NUM>, <NUM>µmol) were dissolved in THF/<NUM> K<NUM>CO<NUM> (<NUM>:<NUM>, <NUM>) and degassed, backfilling with nitrogen three times. The mixture was heated (<NUM> block temperature) with stirring for <NUM>. The reaction was cooled, diluted with DCM (<NUM>), washed with water (<NUM>), passed through a hydrophobic frit and concentrated in vacuo. The crude product was purified by preparative HPLC, Basic, <NUM>-<NUM> MeCN in Water to afford the title compound (<NUM>, <NUM>µmol, <NUM> % yield) as a dark red solid.

<NUM> NMR (<NUM>, Chloroform-d) δ <NUM> (t, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> (dt, J = <NUM>, <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). <NUM> NMR in CDCl3 <NUM>-<NUM>-prep2 was consistent with product structure at <NUM>% purity. LCMS, Basic, <NUM>-70B-prep, m/z <NUM> [M+H]+ at <NUM>, <NUM>% purity @ <NUM>. Contains <NUM> % CMO3 by LC @ <NUM>.

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthrolin e-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and (<NUM>-(morpholinomethyl)phenyl)boronic acid (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was added and the mixture again de-gassed then the whole heated to <NUM> (block temp, pre-heated). After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> Buchi FlashPure, pre-adsorbed, <NUM>-<NUM>% [<NUM>:<NUM> (<NUM>:<NUM> THF:DCM): <NUM> NH<NUM> in MeOH] in (<NUM>:<NUM> THF:DCM)) gave two cuts of moderately pure product. The centre of the product band was evaporated and taken up in MeCN (<NUM>). After ~<NUM> hr, this was filtered and the solid discarded. Meanwhile, material from the edge of the product band was evaporated and re-slurried from iso-hexanes. This material was combined with the MeCN liquors from the above batch and the resultant purified by column chromatography (<NUM> RediSep Gold, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM). The central cut of this band was evaporated to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM> (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Chloroform-d) δ <NUM> (t, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (app p, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>).

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthro line-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and <NUM>-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)pyrrolidine (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> RediSep Gold, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water), loading in DMSO) to afford product in better but still unsatisfactory purity. The residue was re-purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (br s, <NUM>), <NUM> (dt, J = <NUM>, <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (p, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>).

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and <NUM>-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)piperidine (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> RediSep Gold, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (p, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>).

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and N-ethyl-N-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)ethanamine (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> BuchiFlashPure, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (p, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J= <NUM>, <NUM>).

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroli ne-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and N-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)cyclopentanamine (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> BuchiFlashPure, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, Hz, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (p, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> -<NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), CH<NUM>NHCH not observed.

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroli ne-<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and <NUM>-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)azepane (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> Buchi FlashPure, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (p, J= <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>).

A stirred mixture of <NUM>-bromo-<NUM>,<NUM>-bis(<NUM>-morpholinopropyl)-<NUM>-((<NUM>-(pyrrolidin-<NUM>-yl)ethyl)amino)benzo[lmn][<NUM>,<NUM>]phenanthroline -<NUM>,<NUM>,<NUM>,<NUM>(<NUM>,<NUM>)-tetraone (<NUM>, <NUM> mmol) and <NUM>-methyl-<NUM>-(<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)benzyl)piperazine (<NUM>, <NUM> mmol) in dioxane (<NUM>) was treated with potassium carbonate (<NUM>µL of a <NUM> aq solution, <NUM> mmol) and de-gassed. S-Phos Pd G3 (<NUM>, <NUM>µmol) was charged, the mixture again de-gassed and the whole heated to <NUM>. After <NUM> hr, the mixture was allowed to cool then diluted with water (<NUM>) and sat aq NaHCOs (<NUM>) and extracted with DCM (<NUM> × <NUM>). The combined organics were dried over Na<NUM>SO<NUM> and evaporated. Column chromatography (<NUM> BuchiFlashPure, <NUM>-<NUM>% (<NUM>:<NUM> DCM: <NUM> NH<NUM> in MeOH) in DCM, loading in DCM) gave product in moderate purity. The residue was purified by reverse phase column chromatography (<NUM> Reveleris C-<NUM>, <NUM>-<NUM>% (<NUM> NH<NUM> in MeOH) in water, loading in DMSO) to afford the product as a bright red glassy solid (<NUM>, <NUM>%).

LCMS: Found m/z <NUM>: (C<NUM>H<NUM>N<NUM>O<NUM> (MH+) requires <NUM>) @ <NUM>. <NUM>H NMR (<NUM>, Methylene Chloride-d<NUM>) δ <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>).

The CellTiter <NUM>® AQueous One Solution Cell Proliferation Assay (Invitrogen) is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The CellTiter <NUM>® AQueous One Solution Reagent contains a novel tetrazolium compound [<NUM>-(<NUM>,<NUM>-dimethylthiazol-<NUM>-yl)-<NUM>-(<NUM>-carboxymethoxyphenyl)-<NUM>-(<NUM>-sulfophenyl)-<NUM>-tetrazolium, inner salt; MTS] and an electron coupling reagent (phenazine ethosulfate; PES). PES has enhanced chemical stability, which allows it to be combined with MTS to form a stable solution. The MTS tetrazolium compound (Owen's reagent) is bioreduced by cells into a colored formazan product that is soluble in tissue culture medium. Assays are performed by adding a small amount of the CellTiter <NUM>® AQueous One Solution Reagent directly to culture wells, incubating for <NUM>-<NUM> hours and then recording the absorbance at <NUM> with a <NUM>-well plate reader. The quantity of formazan product as measured by absorbance at <NUM> is directly proportional to the number of living cells in culture. The kit was used as per the manufacturers' instructions. After <NUM>-hour incubation with each example compound in MIA-PACA2 cells, the cell proliferation of each sample was measured using the MTS Cell Titre <NUM> Aqueous One Solution Cell Proliferation Assay (Promega Ltd). The percentage of inhibition was calculated against the mean of the DMSO treated controls samples.

Mice aged <NUM>-<NUM> weeks weighing approximately <NUM>-<NUM> were implanted for the study and purchased from Charles River. The pancreatic tumour cell implantation procedure involved MIA-PACA2 cells (<NUM>×<NUM><NUM> in Matrigel) being implanted subcutaneously using a <NUM>-gauge needle onto the rear flank of the mice. Parameters evaluated include: tumour size and animal bodyweight. Tumour volume was measured three times weekly and bodyweight at least <NUM> times weekly. Allocation to treatment groups was done randomly when tumours reached approximately <NUM><NUM> for animals in the efficacy study. Animals (female athymic nude mice bearing MIA-PACA2 tumours) were IV dosed for <NUM> days, twice weekly, at doses of <NUM> and <NUM>/kg for C1, and at doses of <NUM> and <NUM>/kg for Example <NUM>, on account of its <NUM>-fold greater cellular potency. Each group comprised <NUM> animals. All protocols used in this study were approved by the appropriate Animal Welfare and Ethical Review Board, and all procedures were carried out under the guidelines of the UK Animal (Scientific Procedures) Act <NUM>. Results are shown in the tables below and <FIG>.

The graph in <FIG> shows the Xenograft data in the MIA-PACA2 model, after <NUM> days IV administration, followed by <NUM> days measurement (performed by AXIS BioServices). The data shown is mean ± SD n=<NUM> up to day <NUM> and n=<NUM> to the end of the study. The data shows that the compound of Example <NUM> inhibited the growth of the pancreatic tumour and reduced the tumour's size considerably more than the comparative compound C1 or the known anti-cancer drug, Gemcitabine, did, even at a once-weekly dosing regimen. Furthermore, Example <NUM> and the dosing schedules were well-tolerated showing no sign of adverse effects. The starting tumour volumes were <NUM><NUM>. Example <NUM> was active in both of the dosage regimens examined, <NUM> x weekly and <NUM> x weekly, both at a <NUM>/kg dose. Both had <NUM>/<NUM> complete regression in tumour volume at the end of the dosing period. In the complete regression cohorts, tumours have completely disappeared and no regrowth seen after <NUM> days post-dosing. A minority of tumours in the C1 and Example <NUM> groups do not show complete regression, but do show reductions in tumour growth, leading to consistently smaller volumes than in the vehicle control groups.

The CyQUANT XTT Cell Viability Assay (Invitrogen) is a complete, optimized assay that generates a consistent colorimetric detection of viable mammalian cells. The assay kit consists of two reagents, XTT Reagent (<NUM>,<NUM>-bis-(<NUM>-methoxy-<NUM>-nitro-<NUM>-sulfophenyl)-<NUM>-tetrazolium-<NUM>-carboxanilide) and Electron Coupling Reagent. XTT Reagent is used to assess cell viability as a function of cellular redox potential, and the electron coupling reagent improves the dynamic range of the assay. The kit was used as per the manufacturers' instructions.

Claim 1:
A compound of Formula I:
<CHM>
L is in the meta or para position of the phenyl ring;
L is (CH<NUM>)<NUM>-<NUM> and R<NUM> is selected from the group consisting of nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl and NR<NUM>R<NUM>, or L is (CH<NUM>)<NUM>-<NUM>NH and R<NUM> is a C<NUM>-<NUM>cycloalkyl; wherein the nitrogen-containing <NUM>-<NUM> membered heterocycloalkyl is optionally substituted with a C<NUM>-C<NUM> alkyl group;
R<NUM> is a straight chain C<NUM>-<NUM> alkanediyl;
R<NUM> is H;
R<NUM> is a straight chain C<NUM>-<NUM>-alkanediyl;
R<NUM> and R<NUM> are independently C<NUM>-<NUM> alkyl;
X is NR<NUM><NUM>, wherein the R<NUM> groups together with the N atom to which they are attached form pyrrolidin-<NUM>-yl; and
salts, hydrates and solvates thereof.