WO2009127059A1

WO2009127059A1 - Complexes of isogranulatimide and granulatimide with a cyclodextrm, formulations and their use for the treatment of cancer

Info

Publication number: WO2009127059A1
Application number: PCT/CA2009/000495
Authority: WO
Inventors: Gilles Tremblay; Aida Kalbakji; Mario Filion
Original assignee: Alethia Biotherapeutics Inc
Current assignee: Alethia Biotherapeutics Inc
Priority date: 2008-04-15
Filing date: 2009-04-15
Publication date: 2009-10-22
Anticipated expiration: 2010-10-15

Abstract

Formulations of amorphous complexes of compounds of formula (I), such as isogranulatimide or granulatimide, with at least one cyclodextrin are disclosed. Said formulations further comprise a water- soluble compound for inducing osmosis, such as dextrose and/or sodium chloride. Also provided are methods for preparing the aforementioned formulations and methods for delivery in in vitro and in vivo applications. The formulations may be suitable for cancer indications where it is desired to enhance the cytotoxic effectiveness of anti-cancer drugs such as DNA damaging or anti-mitotic agents. In addition, these formulations may be suitable for cancer indications where it is desired to increase the effectiveness of radiotherapy. In the compounds of formula (I), W is defined as being a group of formula (i) or (ii) in which K, E and T are independently N, CR or CZ; Q is NR², O, S or C(R)₂; R¹ is R, RCO- or ArCO-; X and Y are O, H, OH or H₂ and R, Z, R² and Ar are as defined in claim 1.

Description

Complexes of isogranulatimide and granulatimide with a cyclodextrm, formulations and their use for the treatment of cancer

BACKGROUND PCT WO 99/47522 describes novel granulatimide compounds, isogranulatimide compounds, derivatives thereof and pharmaceutical formulations thereof.

Given the low solubility in water of isogranulatimide, granulatimide, and their derivatives, the initial studies describing the preliminary characterization of the molecules were carried out in organic solvents such as dichloromethane or dimethyl sulfoxide (DMSO). The studies described the discovery of isogranulatimide (Roberge et ah, (1998) Cancer Res. 58, 5701 5706), the determination of an improved synthesis (Piers et ah, (2000) J. Org. Chem. 65, 530 535), and the identification of the G₂ checkpoint target, Chkl (Jiang et ah, (2004) MoI. Cancer Ther. 3, 1221 1227). Despite the advances achieved, the utilization of isogranulatimide, granulatimide, and their derivatives in animal models and eventually, human subjects will be limited by the presence of high concentrations of organic solvents that have low maximum tolerated doses in vivo.

SUMMARY

This invention is based, in part, on the discovery that a complexation of granulatimide and/or isogranulatimide and/or derivatives thereof may be made with a cyclodextrin. Furthermore, it was discovered that such complexations have improved solubility and bioavailability. Such complexation may take place in the presence of dextrose. The efficacy of such compounds and complexes may be improved by the formulations and the methods described herein.

In accordance with a first embodiment, there is provided a pharmaceutical composition including a compound of formula I complexed with at least one cyclodextrin selected from: a hydrophilic β-cyclodextrin derivative; a hydrophobic β-cyclodextrins derivative; an ionaizable β-cyclodextrins derivative; an α-cyclodextrin; and a γ-cyclodextrin, wherein the compound of formula I has the structure:

W may be selected from the group consisting of formula (i) or (ii) wherein the structures may be as follows:

in which K, E and T may be independently selected from the group consisting of: N, CR, and CZ, and wherein R and Z may be as defined below:

and in which K and E may be independently selected from the group consisting of: N, CR and CZ, and wherein R, Z and Q may be as defined below.

R¹ may be selected from the group consisting of: R; RCO-; ArCO-; and, ArCH₂-, wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z may be defined as below.

R² may be selected from the group consisting of: H; a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂; -NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CHO; -COR³; -CONH₂; -CONHR³; -CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and - SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z. Each R may be independently selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂; -NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R³; -CHO; -COR³; -CONH₂; -CONHR³; -CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and -SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group.

Each Z may be independently selected from the group consisting of: H; -OH; -OR; -O₂CR; -SH; -SR; -SOCR; -NH₂; -NHR; -NH(R)₂; -NHCOR; NRCOR; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R; -CHO; -COR; -CONH₂; -CONHR; -CON(R)₂; -COSH; -COSR; -NO₂; -SO₃H; -SOR; and, -SO₂R.

Q may be selected from the group consisting of: NR²; O; S; and C(R)₂. X and Y may be independently selected from the group consisting of: O; H; OH; and H₂.

R¹ may be H or CH₃. Q may be NH. X and Y may be oxygen. W may be a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom. Where E, K and T are present, they may be N or CH.

In accordance with a further embodiment, there is provided a pharmaceutical composition that includes a compound of the formula:

complexed with at least one cyclodextrin selected from: a hydrophilic β-cyclodextrin derivative; a hydrophobic β-cyclodextrins derivative; an ionaizable β-cyclodextrins derivative; a α-cyclodextrin; and a γ-cyclodextrin.

The pharmaceutical compositions, may include a further pharmaceutically acceptable carrier. The pharmaceutical compositions, may be for treating cancer. The pharmaceutical compositions, may be for treating cancer wherein the cancer may be selected from: colon and ovarian.

The pharmaceutical compositions, may be for use in a method of treatment of the human or animal body. The pharmaceutical compositions, may have a volume-weighted mean particle size of less than 5 μm. The pharmaceutical compositions, may have a volume-weighted mean particle size within the range of 0.1 μm and 50 μm. The pharmaceutical compositions, may have a volume-weighted mean particle size of less than 10 μm. The pharmaceutical compositions, may have a volume-weighted mean particle size of less than 25 μm. Particle size may be measured using one or more of the methods known in the art. For example, laser diffraction, spectrophotometry (PSS), dynamic light scattering (DLS) or photon correlation spectroscopy (PCS), and single particle optical sizing (SPOS), also known as optical particle counting (OPC). Particle size may be measured using laser diffraction.

The pharmaceutical compositions, may contain a hydrophilic β-cyclodextrin derivative selected from: methylated β-cyclodextrin; hydroxyalkylated β-cyclodextrin; and branched β-cyclodextrin.

The pharmaceutical compositions, may contain a hydrophobic β-cyclodextrin derivative selected from: alkylated β-cyclodextrin; and acylated β-cyclodextrin. The ionaizable β-cyclodextrin derivative may be anionic β-cyclodextrin. The at least one cyclodextrin may comprise 2-hydroxypropyl-β-cyclodextrin (HPCD) .

The pharmaceutical compositions may comprise a water-soluble compound for inducing osmosis. The water-soluble compound for inducing osmosis may be selected from: sodium chloride, mannitol, dextrate, and dextrose. The water-soluble compound for inducing osmosis may be selected from: 0.9% sodium chloride and 5% dextrose.

The pharmaceutical compositions may be used in the manufacture of a medicament for the treatment of cancer. The medicament may sensitize cancer cells to the effects of DNA damaging agents on cancer cells. The pharmaceutical compositions may be used wherein the cancer may be colon or ovarian. The pharmaceutical compositions may be used for the treatment of cancer. The pharmaceutical compositions may be used wherein the cancer may be colon or ovarian.

In accordance with a further embodiment, there is provided a complexation product wherein a cyclodextrin may be complexed with a compound of formula I having the structure:

wherein, W may be selected from the group consisting of formula (i) or (ii) wherein the structures may be as follows:

in which K, E and T may be independently selected from the group consisting of: N, CR, and CZ, and wherein R and Z may be as defined below; and

in which K and E may be independently selected from the group consisting of: N, CR and CZ, and wherein R, Z and Q may be as defined below. R¹ may be selected from the group consisting of: R; RCO-; ArCO-; and, ArCH₂-, wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z may be defined as below.

R² may be selected from the group consisting of: H; a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂; -NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CHO; -COR³; -CONH₂; -CONHR³; -CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and - SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z.

Each R may be independently selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂;

-NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R³; -CHO; -COR³; -CONH₂; -CONHR³;

-CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and -SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group.

Q may be selected from the group consisting of: NR²; O; S; and C(R)₂. X and Y may be independently selected from the group consisting of: O; H; OH; and H₂. R¹ may be H or CH₃. Q may be NH. X and Y may be oxygen. W may be a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom. Where E, K and T are present, they may be N or CH.

In accordance with a further embodiment, there is provided a composition that includes a cyclodextrin complexed with a compound of formula I having the structure:

wherein W may be selected from the group consisting of formula (i) or (ii) wherein the structures may be as follows:

in which K and E may be independently selected from the group consisting of: N, CR and CZ, and wherein R, Z and Q may be as defined below.

R¹ may be selected from the group consisting of: R; RCO-; ArCO-; and, ArCH₂-, wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z may be defined as below. R² may be selected from the group consisting of: H; a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂; -NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CHO; -COR³; -CONH₂; -CONHR³; -CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and - SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar may be an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z.

Each R may be independently selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR³; -O₂CR³; -SH; -SR³; -SOCR³; -NH₂; -NHR³; -NH(R³)₂; -NHCOR³; NRCOR³; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R³; -CHO; -COR³; -CONH₂; -CONHR³; -CON(R³)₂; -COSH; -COSR³; -NO₂; -SO₃H; -SOR³; and -SO₂R³, wherein each R³ may be independently selected from the group consisting of: a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group.

Each Z may be independently selected from the group consisting of: H; -OH; -OR; -O₂CR; -SH;

-SR; -SOCR; -NH₂; -NHR; -NH(R)₂; -NHCOR; NRCOR; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R;

-CHO; -COR; -CONH₂; -CONHR; -CON(R)₂; -COSH; -COSR; -NO₂; -SO₃H; -SOR; and, -SO₂R.

Q may be selected from the group consisting of: NR²; O; S; and C(R)₂. X and Y may be independently selected from the group consisting of: O; H; OH; and H₂; wherein the weight ratio of the compound to cyclodextrin ranges from 1 :10 to 1 :1000, preferably from 1 :50 to 1 :1000, and more preferably from 1 :50 to 1 :500.

R¹ may be H or CH₃. Q may be NH. X and Y may be oxygen. W may be a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom. Where E, K and T, where present, they may be N or CH. In accordance with a further embodiment, there is provided a method of producing a water soluble amorphous pharmaceutical composition. The method involves combining a cyclodextrin and a compound of formula I having the structure:

Q may be selected from the group consisting of: NR²; O; S; and C(R)₂. X and Y may be independently selected from the group consisting of: O; H; OH; and H₂ and wherein the weight ratio of the compound to cyclodextrin ranges from 1:10 to 1:1000, preferably from 1 :50 to 1:1000, and more preferably from 1 :50 to 1 :500.

R¹ may be H or CH₃. Q may be NH. X and Y may be oxygen. W may be a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom. Where E, K and T, where present, they may be N or CH.

The method may include the pharmaceutical composition comprising a compound of the formula:

The method may include the pharmaceutical composition comprising a further pharmaceutically acceptable carrier. The method may include the pharmaceutical composition formulated for intravenous administration.

The method may include the pharmaceutical composition wherein the particles have a volume-weighted mean particle size of less than 5 μm. The method may include the pharmaceutical composition wherein the particles have a volume-weighted mean particle size within the range of 0.1 μm and 50 μm. The method may include the pharmaceutical composition wherein the particles have a volume-weighted mean particle size of less than 10 μm. The method may include the pharmaceutical composition wherein the particles have a volume-weighted mean particle size of less than 25 μm. Particle size may be measured using one or more of the methods known in the art. For example, laser diffraction, spectrophotometry (PSS), dynamic light scattering (DLS) or photon correlation spectroscopy (PCS), and single particle optical sizing (SPOS), also known as optical particle counting (OPC). Particle size may be measured using laser diffraction.

The method may include cyclodextrin selected from one or more of the following: a hydropnilic β-cyclodextrin derivative; a hydrophobic β-cyclodextrin; an ionaizable β-cyclodextrin derivative; an α-cyclodextrin; and a γ-cyclodextrin.

The method may include cyclodextrin selected from one or more of the following: methylated β-cyclodextrin; hydroxyalkylated β-cyclodextrin; branched β-cyclodextrin; alkylated β-cyclodextrin; acylated β-cyclodextrin; anionic β-cyclodextrin; α-cyclodextrin; and a γ-cyclodextrin. The method may include 2-hydroxypropyl-β-cyclodextrin (HPCD).

The method may include the addition of a water-soluble compound for inducing osmosis. For example, the water-soluble compound or inducing osmosis may be isotonic. For example, the water-soluble compound for inducing osmosis may be comprised of one or more of the following: sodium chloride, mannitol, dextrate, and dextrose. For example, the water-soluble compound for inducing osmosis may be selected from: 0.9% sodium chloride and 5% dextrose. The method may include a solution formed in an aqueous medium.

The method may comprise stabilizing the water soluble amorphous pharmaceutical composition with one or more of: ultrasound energy; heat; elevated pressure; and mechanical agitation. For example, wherein stabilizing the water soluble amorphous pharmaceutical composition may include the use of ultrasound energy. For example, wherein stabilizing the water soluble amorphous pharmaceutical composition may include elevation of the water soluble amorphous pharmaceutical composition to a temperature of between about 4O⁰C and about 70⁰C. For example, the mechanical agitation may include stirring.

In accordance with a further embodiment, there are provided uses of the pharmaceutical compositions described herein for the preparation of a medicament for the treatment of cancer. In accordance with a further embodiment, there are provided uses of the pharmaceutical compositions described herein for the treatment of cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures IA and IB are illustrations displaying the chemical structures of isogranulatimide (IA); and the chemical structures of granulatimide (IB).

Figure 2 are graphs displaying the effect of isogranulatimide and topotecan on cell lines TOV-21G and TOV-112D.

Figure 3 is a graph displaying the effect of isogranulatimide and topotecan on the cell line TOV-21G in the presence of DMSO and HPCD.

Figure 4 is a graph displaying the effect of isogranulatimide and irinotecan on the cell line HCT-116 in the presence of DMSO and HPCD.

Figure 5 is a graph displaying the effect of isogranulatimide and topotecan on the cell line TOV-21G.

Figure 6 is a graph displaying the effect of isogranulatimide and topotecan on the cell line HCT-116.

DETADLED DESCRIPTION Granulatimide, isogranulatimide, derivatives thereof and pharmaceutical formulations thereof are described in PCT WO 99/47522. These compounds include the naturally occurring compounds, granulatimide and isogranulatimide, which may be in purified or partially purified form, including extracts containing these compounds taken from naturally occurring sources (for example, Didemnum granulatum). Alternatively, these compounds may be synthesized or partially synthesized from purified extracts. These compounds may be useful as a cytotoxic agents, as a protein kinase inhibitors, or as G2 checkpoint inhibitors. Furthermore, the compounds may be used to sensitize cells to the effects of DNA damaging agents; and the use of such compounds in the formulation of agents, including medicaments.

Cyclodextrins are a family of cyclic oligosaccharides, which are linked via α-1,4 linkages. Depending upon the number of units, cyclodextrins can be classified as α-cyclodextrin, a six- membered oligosaccharide; β-cyclodextrin, a seven-membered oligosaccharide; and γ- cyclodextrin, an eight-membered oligosaccharide. As described generally in Davis, M.E. and

M.E. Brewster. (2004) Cyclodextrin-based pharmaceutics: past, present and future. Nature

Reviews Drug Discovery 3, 1023-1035, cyclodextrins may include glucosyl-β-cyclodextrin, maltosyl-β-cyclodextrin, dimaltosyl-β-cyclodextrin, carboxymethyl-α-cyclodextrin, 2- hydroxypropyl-γ-cyclodextrin, sulphobutylether-β-cyclodextrin, randomly methylated-β- cyclodextrin, and 2-hydroxypropyl-β-cyclodextrin.

As used herein the term "skeleton" refers to a group of atoms that make up the main atomic chain or ring of a chemical moiety. The main atomic chain or ring does not include atoms that are substituents on those atoms. For example, methane has a main atomic chain consisting of 1 carbon atom and 4-methyl-pentane has main atomic chain consisting of 5 carbon atoms. A skeleton containing one to ten carbon atoms contains 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms in the main atomic chain or ring, but the chemical moiety that comprises the skeleton, may have more than 10 atoms in total. The chemical moiety may have more than 10 carbon atoms in total. For example, methane, chloromethane, n-butane, tert-butane, 3-methyl-decane, and

1 -ethyl, 5-chloro-cyclodecane are all examples of chemical moieties that each comprise a skeleton having 1 to 10 carbon atoms.

Conjugations

Persons skilled in the art would be capable of conjugating the compounds described herein to various functional groups to further improve solubility, biocompatibility, delivery, targeting and thus the overall therapeutic index. Such modifications may allow a person skilled in the art to conjugate the compound to functional groups such that the antimitotic activity is enhanced.

The functional groups conjugated to the compound may be a biological delivery and targeting molecule. For the purposes of this invention, biological delivery and targeting molecules are those that bind to a specific biological substance or site. The biological substance or site is the intended target of the biorecognition molecule that binds to it, enabling the delivery of the compound to the tissue or cells of interest (for example, neoplastic colon or neoplastic ovary). Targeting of the compound may be accomplished by conjugating it to a biological delivery and targeting molecule. Examples of biological delivery and targeting molecules are described below.

A ligand may function as a biological delivery and targeting molecule by selectively binding or having a specific affinity for another substance. A ligand is recognized and bound by a specific binding body or binding partner, or receptor. Examples of ligands suitable for targeting are antigens, haptens, biotin, biotin derivatives, lectins, galactosamine and fucosylamine moieties, receptors, substrates, coenzymes and cofactors among others.

A ligand may include cancer and tumor antigens such as alpha-fetoproteins, prostate specific antigen (PSA) and CEA, cancer markers and oncoproteins, among others. Other substances that can function as ligands for delivery and targeting are certain steroids, prostaglandins, carbohydrates, lipids, certain proteins or protein fragments (i.e. hormones, toxins), and synthetic or natural polypeptides with cell affinity. Ligands may also include various substances with selective affinity for ligators that may be produced through recombinant DNA, genetic and molecular engineering.

Another type of delivery and targeting molecule may be an antibody, which is defined to include all classes of antibodies, monoclonal antibodies, chimeric antibodies, Fab fractions, fragments and derivatives thereof. Other delivery and targeting molecules include enzymes, especially cell surface enzymes such as neuraminidases, plasma proteins, avidins, streptavidins, chalones, cavitands, thyroglobulin, intrinsic factor, globulins, chelators, surfactants, organometallic substances, staphylococcal protein A, protein G, cytochromes, lectins, certain resins, and organic polymers.

Delivery and targeting molecules may also include various substances such as any proteins, protein fragments or polypeptides with affinity for the surface of any cells or tissues to be targeted by the compound. These proteins may be produced through recombinant DNA, genetic and molecular engineering techniques know in the art. Any suitable membrane transfer proteins to facilitate the transfer of the compound to the target cell interior may be of particular use.

Delivery and targeting molecules may not only be desirable for delivery to a target cell or tissue but may also to facilitate the transport of the compound into a cell. One such example is U.S. Pat. No 6,204,054, which describes the use of transcytosis vehicles and enhancers capable of transporting physiologically-active agents across epithelia, endothelia and mesothelia containing the GP60 receptor. The GP60 receptor has been implicated in receptor-mediated transcytosis of albumin across cell barriers. U.S. Pat. No 6,204,054 exploits GP60 receptor-mediated transcytosis for the transport of physiologically-active agents which do not naturally pass through epithelia, endothelia and mesothelia via the GP60 system. A compound can be coupled to albumin, albumin fragments, anti-GP60 polyclonal and monoclonal antibodies, anti-GP60 polyclonal and monoclonal antibody fragments, and GP60 peptide fragments to facilitate transport into the cell.

The conjugation to a functional group may also improve other properties of the compound. Such functional groups are often termed drug carriers and can improve the stability, solubility or biocompatibility of the drug being carried.

For example alternative or additional alterations to improve solubility of the compound may be achieved by conjugating the compound to a peptide polymer. U.S. Pat. Publication No. 2001041189 describes the use of polypeptides (containing glutamic acid and aspartic acid, or glutamic acid/alanine, or glutamic acid/asparagine, or glutamic acid/glutamine, or glutamic acid/glycine) as conjugated to drugs to act as carriers to improve the solubility of the drugs and/or their therapeutic efficacy in vivo. One such drug exemplified in US 2001041189 is the poorly soluble paclitaxel. U.S. Pat. No. 4,675,381 describes a polyaspartate and/or polyglutamate polymer as a drug carrier. This patent suggests the use of polyaspartate and/or polyglutamate polymers as drug carriers wherein the drug is encapsulated or incorporated in the matrix of the polymer. Similarly, U.S. Pat. No. 5,087,616 describes the use of a biodegradable polymeric carrier to which one or more cytotoxic molecules, such as daunomycin is conjugated. The biodegradable polymeric carrier is specified to be, for example, a homopolymer of polyglutamic acid. Also, U.S. Pat. No. 4,960,790 describes the anti-tumor agent taxol covalently conjugated to an amino acid (glutamic acid). U.S. Pat. No. 5,420,105 describes the use of polypeptide carriers that are capable of binding one drug or multiple drugs. In U.S. Pat. No. 5,420,105 the polypeptide carrier may be further attached to a targeting or delivery protein, such as an antibody or ligand capable of binding to a desired target site in vivo.

U.S. Pat. No. 6,127,349 describes the use of phospholipids to improve the solubility of the therapeutic agents (steroids, peptides, antibiotics and other biologically active agents and pharmaceutical formulations) and to improve their bio-availability. Similarly, fatty acids could be conjugated to the compound in order to stabilize the activity of the anti-angiogenic substances. U.S. Pat. No. 6,380,253 describes the conjugation of anti-angiogenic substances (proteins - angiostatin and endostatin etc.) to cis unsaturated fatty acids or polyunsaturated fatty acids to potentiate and stabilize the activity of the anti-angiogenic substances. Other suitable drug carriers include Polyethylene glycol (PEG) and related polymer derivatives. Such drug-PEG conjugates have been described as improving the circulation time (prolong serum half-life) before hydrolytic breakdown of the conjugate and subsequent release of the bound molecule thus increasing the drugs efficacy. U.S. Pat. No. 6,214,966 describes the use of PEG and related polymer derivatives having weak, hydrolytically unstable linkages near the reactive end of the polymer to conjugate to drugs such as proteins, enzymes and small molecules.

Alternatively, EP 1082105 (WO9959548) describes the use of biodegradable polyester polymers as a drug delivery system to facilitate controlled release of the conjugated drug.

As another alternative the compound may be conjugated to another pharmaceutically active compound to enhance the therapeutic effect on the target cell or tissue by delivering a second compound with a similar anti-mitotic effect or a different activity altogether. US 6,051,576 describes the use of co-drug formulations by conjugating two or more agents via a labile linkage to improve the pharmaceutical and pharmacological properties of pharmacologically active compounds.

Two distinct types of conjugations are described as follows. One type of conjugation can be through noncovalent or attractive binding as with an antigen and antibody or biotin and avidin. Noncovalent coupling is binding between substances through ionic or hydrogen bonding or van der waals forces, and/or their hydrophobic or hydrophilic properties. Alternatively, conjugation may be through covalent, electron-pair bonds or linkages. Many methods and agents for covalent conjugation (or crosslinking) are known and, with appropriate modification, can be used to conjugate the desired substances to the compound. Where stability is desired, linkages may be amide bonds, peptide bonds, ether bonds, and thio ether bonds, among others.

For example, the in vitro activity of isogranulatimide in ovarian and colon cancer cells in combination with DNA damaging agents such as the topoisomerase I inhibitors topotecan and irinotecan is described. Topoisomerase I is an essential human enzyme that is required for the unwinding of double-stranded DNA during processes such as DNA synthesis or transcription. The alkaloid captothecin, from which the clinically approved derivatives topotecan and irinotecan were derived, is a potent inhibitor of topoisomerase I that causes double-stranded DNA breaks. Topotecan and irinotecan are used as anti-cancer agents in several solid tumor indications (Pommier, Y. (2006) Nat. Rev. Cancer 6, 789 802). Other DNA damaging agents that could be used include topoisomerase II inhibitors such as etoposide and doxorubicin also function by inhibiting proper DNA unwinding (Nitiss, J.L (2002) Curr. Opin. Investig. Drugs 3, 1512 1516), DNA alkylating agents such as mitomycin C (Tomasz, M. and Palom, Y. (1997) Pharmacol. Ther. 76, 73 87); and DNA damage by UV or gamma irradiation. The first experiments were conducted with isogranulatimide resuspended in 100% DMSO (see Example 1) and show that complexation of isogranulatimide in 10% hydroxy-propyl β-cyclodextrin in the presence of 5% dextrose improves the efficacy of the molecules by close to 10-fold (see Examples 2, 3, and 4).

Pharmaceutical Compositions

Further modifications of the pharmaceutical compositions and conjugations set out herein may be made as set out below to enhance the properties of the pharmaceutical compositions. Humans, and other animals, in particular, mammals, suffering from proliferative diseases, and other similar conditions may be treated by administering an effective amount of one or more of the above- identified pharmaceutical compositions or a pharmaceutically acceptable derivative or salt thereof in a pharmaceutically acceptable carrier or diluent. The active materials may be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, or subcutaneously.

As used herein, the term pharmaceutically acceptable salts or complexes may refer to salts or complexes that retain the desired biological activity of the above-identified compounds and exhibit minimal undesired toxicological effects. Nonlimiting examples of such salts are (a) acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid; (b) base addition salts formed with polyvalent metal cations such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, nickel, cadmium, sodium, potassium, and the like, or with an organic cation formed from N,N-dibenzylethylene-diamine, D- glucosamine, ammonium, tetraethylamrnonium, or ethylenediamine; or (c) combinations of (a) and (b); e.g., a zinc tannate salt or the like.

The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount suffcient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated. A dose of the active compound for the above-mentioned conditions may be in the range from about 0.5 to 500 mg/kg, or 1 to 100 mg/kg per day. The effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered. If the derivative exhibits activity in itself, the effective dosage can be estimated as above using the weight of the derivative, or by other means known to those skilled in the art.

The compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing 1 to 3000 mg, preferably 5 to 500 mg of active ingredient per unit dosage form. An oral dosage of 25-250 mg is often convenient.

The active ingredient may be administered to achieve peak plasma concentrations of the active compound of about 0.1 to 100 μM, or about 1-10 μM. This may be achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or administered as a bolus of the active ingredient.

The concentration of active compound in the drug composition may depend on absorption, distribution, inactivation/metabolism, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens may be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application may include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of toxicity such as sodium chloride or dextrose.

If administered intravenously, carriers may be physiological saline or phosphate buffered saline (PBS). The active compound can also be administered through a transdermal patch. Methods for preparing transdermal patches are known to those skilled in the art. For example, see Brown L., and Langer R., Transdermal Delivery of Drugs, Annual Review of Medicine, 39:221-229 (1988).

The active compounds may be prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid.

Liposomal suspensions may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in US patent 4,522,811. For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine stearoyl phosphatidyl choline, arachadoyl phosphatidy choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound or its monophosphate, and/or triphosphate derivatives are then introduced into the container. The container is then swirled by hand to free the lipid aggregates, thereby forming the liposomal suspension.

Oral compositions may include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound may be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.

The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or other enteric agents.

Methods are known for encapsulating compositions (such as in a coating of hard gelatin) for oral administration are well known in the art (Baker, Richard, Controlled Release of Biological Active Agents, John Wiley and Sons, 1986). Suitable pharmaceutically acceptable carriers for parenteral application, such as intravenous, subcutaneous or intramuscular injection, include sterile water, physiological saline, bacteriostatic saline (saline containing 0.9 mg/ml benzyl alcohol) and phosphate-buffered saline. The active compound or pharmaceutically acceptable salt or derivative thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

The active compound or pharmaceutically acceptable derivatives or salts thereof can also be administered with other active materials that do not impair the desired action, or with materials that supplement the desired action.

Particle Size Measurement Techniques

Particle size spectrophotometry (PSS) uses visible and ultra-violet light to differentiate between very similar particle sizes in multi-modal distributions. Particle size spectrophotometry measures light scattered at a fixed angle as a function of its wavelength. By using a wide range of wavelengths of ultra-violet to visible light (for example, 190 run to 1100 nm) it is possible to measure a wide range of particles from 5 nm to 15 μm.

Dynamic light scattering (DLS) or photon correlation spectroscopy (PCS) is capable of precision particle size measurements from 0.5 nm to 6 μm. This method measures the intensity of light scattered in a particular direction by particles in a sample. The intensity of scattered light changes with time, due to the Brownian motion of particles in the suspension. Generally, instruments obtain a correlation factor from the intensity versus time profile. The exponentially decaying correlation function is related to diffusion coefficients, and using the Stokes-Einstein equation and these coefficients the instrument can calculate the particle radius. It is an absolute measurement (i.e. knowledge of the particle composition is not needed) and can measure particles ranging in size from 0.5 nm to 6 μm.

Laser Diffraction is a method for measuring particle sizes in a suspension. This method is usually used to measure the sizes of particles of a few μm to 1000's of μm. A laser is used to scatter light off particles in a dilute suspension. A lens focuses the scattered light on a detector and the intensity of light at various angles on the detector can be correlated using the Mie scattering effect to determine the particle size.

Single particle optical sizing (SPOS), also known as optical particle counting (OPC) is capable of identifying outlier particle sizes in distributions. Particles are passed one at a time past a laser where light scattering or light blockage is counted by a detector. This method is capable of measuring particles ranging in size from 0.5 μm to 2500 μm.

The size of a particle determined by a given technique may vary from another technique, due to the use of the equivalent sphere approximation. Equivalent sphere approximation is often used to derive a single diameter that is indicative of the size, and depending on the technique may provide different size distributions. Different techniques measure different particle properties (for example, sedimentation rate, light scattering patterns, or projected image size) and it is accordingly important to consider the non-sphericity and statistical representations when comparing the results of different methods. For example, a non-spherical particle may affect a measurement in different ways depending on the physical property being measured in order to derive the particle size. Accordingly, particle sizing standard reference materials (which tend to be spherical) may be used so that similar results can then be generated by multiple techniques or alternatively, particle sizing standard reference materials may be used to optimize settings and analysis. The statistical representations will likely vary when different techniques are used. Such statistical representations may apply different weightings to the particles within the distribution. For example, one technique may measure the number of particles of a given size whereas another may measure the mass of particles and may affect the shape reported size distribution and the technique's sensitivity to changes in the distribution width.

Ultimately, it is unlikely that any particular particle size determination method is superior in making particle size determinations than another. Furthermore, the focus is likely best placed on whether the measured parameter relates to the application being developed. In many applications, such as in the production of pharmaceuticals, it is important to understand how the mass or volume of a material is distributed between particles of different sizes. Accordingly, laser diffraction is widely used in the pharmaceutical industry.

EXAMPLES EXAMPLE 1 The ability of isogranulatimide to enhance the cytotoxic effect of topotecan was measured in ovarian cancer cells. Two human ovarian cancer cell lines derived from chemotherapy-naive patients were utilized that have been previously characterized in terms of their morphology, tumorigenicity, and global expression profiles. In addition, these analyses revealed that these cell lines were excellent models for in vivo behavior of ovarian tumors in humans (Provencher D.M. et al, (2000) In Vitro Cell. Dev. Biol Anim. 36, 357 361; Samouelian V. et ah, (2004) Cancer Chemother. Pharmacol. 54, 497 504). These cell lines are designated TOV-21G and TOV-112D. A lO mg/ml stock solution of isogranulatimide was prepared in 100% DMSO. In order to measure the efficacy, 4,000 cells/well TOV-21G or TOV-112D cells were seeded in 48-well plates and allowed to adhere overnight at 37°C in 5% CO₂. The cells were treated with 10 or 5 nM topotecan for 16 hours to induce DNA damage. Following this treatment, the cells were incubated for 3 days in the presence of 5 μM isogranulatimide after which the cell number was determined spectrophotometrically by MTT assay at 570 nm (Mosmann, T. (1983) J. Immunol. Methods 65, 55 63). As shown in Figure 2 (left panel, TOV-21G), treatment of TOV-21G cells with topotecan resulted in approximately 50% cell death. Incubation of the cells with isogranulatimide following the topotecan resulted in a further 50% reduction in the cell number. Furthermore, the same percentage of cell death was achieved in the presence of isogranulatimide at both the high and low dose of topotecan. This demonstrates that lower concentrations of the toxic chemotherapeutic compound can be used and exemplifies the chemosensitization effect of isogranulatimide. Very similar results were obtained in the TOV-112D cell line (see Figure 2, right panel, TOV-112D) or in the presence of another chemotherapeutic, doxorubicin. The data represents the mean of at least 2 experiments conducted separately ±SEM. These results represent the first demonstration of isogranulatimide chemosensitization activity in ovarian cancer cells.

EXAMPLE 2

In this example, a comparative study was performed to ascertain the efficacy of isogranulatimide formulated in HPCD compared to isogranulatimide resuspended in DMSO. The complexation reaction that resulted in the formulation of isogranulatimide in cyclodextrins can be accomplished as follows: 5 grams of solid HPCD (Sigma™ #H107) was resuspended in sterile water containing 5% dextrose (D5W, Baxter Corporation™ #JB0062) until complete dissolution after which the volume was increased to exactly 50 ml. This solution was filter sterilized through a 0.22 micron filter. Thirty milligrams of solid isogranulatimide was weighed and added to 3 ml HPCD/dextrose solution in a glass amber crimp vial. Following brief vortexing of the mixture, the vial was incubated for 15 minutes in a Branson 1200 sonicating water bath preheated to 55⁰C. This sonication was repeated three times until a homogenous suspension was achieved. Therefore, this procedure resulted in a 10 mg/ml stock solution of isogranulatimide in an aqueous milieu that completely lacked any organic solvents. Thus, a greater solubility was achieved with the cyclodextrin. The isogranulatimide-HPCD formulation was stored at 4⁰C in the dark.

The chemosensitizing activity of the isogranulatimide-HPCD formulation was tested in ovarian cancer cells and directly compared with isogranulatimide resuspended in DMSO. As described in Example 1, 4,000 cells/well TOV-21G cells were seeded in 48-well plates and allowed to adhere overnight at 37°C in 5% CO₂. The cells were treated with 10 nM topotecan for 16 hours. Following this treatment, the cells were incubated for 3 days in the presence of isogranulatimide at the indicated concentrations after which the cell number was determined spectrophotometrically by MTT assay at 570 nm. As shown in Figure 3, the isogranulatimide that was complexed with HPCD was significantly more active than the compound resuspended in DMSO. The maximum percentage of inhibition by isogranulatimide was achieved at the lowest concentration used in this example. There was no effect on the proliferation of the cells following 3 days treatment with the HPCD/dextrose vehicle alone. The data represents the mean of at least 2 experiments conducted separately ±SEM.

This example demonstrates that the water-insoluble isogranulatimide can be formulated in aqueous buffers in the presence of cyclodextrin and this formulation results in increased biological activity of the compound as a chemosensitizer of DNA damaging agents.

EXAMPLE 3

The activity of isogranulatimde was also assessed in a colon carcinoma cell line (ATCC #CCL-247). The experiment of Example 2 was repeated with the colon cancer cell line under very similar conditions except irinotecan replaced topotecan as the DNA damaging agent. As was observed with the ovarian cancer cell lines, isogranulatimide was more active when formulated in cyclodextrin (see Figure 4). The data represents the mean of at least 2 experiments conducted separately ±SEM.

EXAMPLE 4

The conditions of Examples 2 and 3 were repeated in this Example but using lower concentrations of isogranulatimide-HPCD formulation. When compared with isogranulatimide that was resuspended in DMSO (see Figure 2), the results shown in Figures 5 and 6 illustrate that enhanced cell death was observed in both ovarian (Figure 5) and colon (Figure 6) cancer cells at concentrations as low as 100 nM isogranulatimide when treated in combination with a DNA damaging agent. This demonstrates that the activity is greatly enhanced when the compound is formulated with cyclodextrins.

Although various embodiments of the invention are disclosed herein, many adaptations and modifications may be made within the scope of the invention in accordance with the common general knowledge of those skilled in this art. Such modifications include the substitution of known equivalents for any aspect of the invention in order to achieve the same result in substantially the same way. Numeric ranges are inclusive of the numbers defining the range. The word "comprising" is used herein as an open-ended term, substantially equivalent to the phrase "including, but not limited to", and the word "comprises" has a corresponding meaning. As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a thing" includes more than one such thing. Citation of references herein is not an admission that such references are prior art to the present invention. Embodiments of the invention include all examples and variations substantially as hereinbefore described and with reference to the examples and drawings.

Claims

CLAIMS:

1. A pharmaceutical composition, the composition comprising a compound of formula I complexed with at least one cyclodextrin selected from: a hydrophilic β-cyclodextrin derivative; a hydrophobic β-cyclodextrins derivative; an ionaizable β-cyclodextrins derivative; an α-cyclodextrin; and a γ-cyclodextrin, wherein the compound of formula I has the structure:

wherein:

W is selected from the group consisting of formula (i) or (ii) wherein the structures are as follows:

or in which K, E and T are independently selected from the group consisting of: N, CR, and CZ, and wherein R and Z are as defined below; and

, or in which K and E are independently selected from the group consisting of: N, CR and CZ, and wherein R, Z and Q are as defined below;

Ri is selected from the group consisting of: R; RCO-; ArCO-; and, ArCH₂-, wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z may be defined as below;

R₂ is selected from the group consisting of: H; a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃; -SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN; -CO₂H; - CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃; -NO₂; -SO₃H; -SOR₃; and - SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z;

R is selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃;

-SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN;

-CO₂H; -CO₂R₃; -CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃; -NO₂; -SO₃H; -SOR₃; and -SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group;

Z is an optional substituent selected from the group consisting of: H; -OH; -OR; -O₂CR; -SH; -SR; -SOCR; -NH₂; -NHR; -NH(R)₂; -NHCOR; NRCOR; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R; -CHO; -COR; -CONH₂; -CONHR; -CON(R)₂; -COSH; -COSR; - NO₂; -SO₃H; -SOR; and, -SO₂R:

Q is selected from the group consisting of: NR₂; O; S; and C(R)₂; and

X and Y are independently selected from the group consisting of: O; H; OH; and H₂.

2. The pharmaceutical composition of claim 1 , wherein Ri is H or CH₃.

3. The pharmaceutical composition of claim 1 or 2, wherein Q is NH.

4. The pharmaceutical composition of any one of claims 1 -3, wherein X and Y are oxygen.

5. The pharmaceutical composition of any one of claims 1 -4, wherein W is a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom; and wherein E, K and T, where present, are N or CH.

6. A pharmaceutical composition, the composition comprising a compound of the formula:

complexed with at least one cyclodextrin selected from: a hydrophilic β-cyclodextrin derivative; a hydrophobic β-cyclodextrins derivative; an ionaizable β-cyclodextrins derivative; a α- cyclodextrin; and a γ-cyclodextrin.

7. A pharmaceutical composition, the composition comprising a compound of the formula:

A pharmaceutical composition, the composition comprising a compound of the formula:

9. A pharmaceutical composition, the composition comprising a compound of the formula:

10. A pharmaceutical composition, the composition comprising a compound of the formula:

11. The pharmaceutical composition of any one of claims 1-10, further comprising a pharmaceutically acceptable carrier.

12. The pharmaceutical composition of any one of claims 1 -11 , for treating cancer.

13. The pharmaceutical composition of any one of claims 1-12, wherein the cancer is selected from: colon and ovarian.

14. The pharmaceutical composition of any one of claims 1 - 13 for use in a method of treatment of the human or animal body.

15. The pharmaceutical composition of any one of claims 1 -14, wherein the particles have a volume-weighted mean particle size of less than 5 μm as measured by laser diffractometry.

16. The pharmaceutical composition of any one of claims 1-15, wherein the hydrophilic β-cyclodextrin derivative is selected from: methylated β-cyclodextrin; hydroxyalkylated β-cyclodextrin; and branched β-cyclodextrin.

17. The pharmaceutical composition of any one of claims 1-15, wherein the hydrophobic β-cyclodextrin derivative is selected from: alkylated β-cyclodextrin; and acylated β- cyclodextrin.

18. The pharmaceutical composition of any one of claims 1-15, wherein the ionaizable β-cyclodextrin derivative is anionic β-cyclodextrin.

19. The pharmaceutical composition of any one of claims 1-16, wherein the at least one cyclodextrin comprises 2-hydroxypropyl-β-cyclodextrin (HPCD).

20. The pharmaceutical composition of any one of claims 1-19, further comprising a water-soluble compound for inducing osmosis.

21. The pharmaceutical composition of claim 20, wherein the water-soluble compound for inducing osmosis is selected from: sodium chloride, mannitol, dextrate, and dextrose.

22. The pharmaceutical composition of claim 20 or 21 , wherein the water-soluble compound for inducing osmosis is selected from: 0.9% sodium chloride and 5% dextrose.

23. A pharmaceutical composition of any one of claims 1 -22, for use in the manufacture of a medicament for the treatment of cancer.

24. The pharmaceutical composition of claim 23, wherein the medicament sensitizes cancer cells to the effects of DNA damaging agents on cancer cells.

25. The pharmaceutical composition of claim 23 or 24, wherein the cancer is colon or ovarian.

26. A use of a pharmaceutical composition of any one of claims 1 -22 for the treatment of cancer.

27. The use of claim 26, wherein the cancer is colon or ovarian.

28. A complexation product wherein a cyclodextrin in the presence of dextrose is complexed with a compound of formula I having the structure:

wherein:

R₁ is selected from the group consisting of: R; RCO-; ArCO-; and, ArCH₂-, wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z may be defined as below;

R₂ is selected from the group consisting of: H; a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃; -SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂;

-NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN; -CO₂H; -CHO; -COR₃; -CONH₂;

-CONHR₃; -CON(R₃)₂; -COSH; -COSR₃; -NO₂; -SO₃H; -SOR₃; and -SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z; R is selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃;

-SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN; -CO₂H; -CO₂R₃; -CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃;

-NO₂; -SO₃H; -SOR₃; and -SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group;

Q is selected from the group consisting of: NR₂; O; S; and C(R)₂; and

29. The complexation product of claim 28, wherein Ri is H or CH₃.

30. The complexation product of claim 28 or 29, wherein Q is NH.

31. The complexation product of any one of claims 28-30, wherein X and Y are oxygen.

32. The complexation product of any one of claims 28-31, wherein W is a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom; and wherein E, K and T, where present, are N or CH.

33. A composition comprising a cyclodextrin complexed with a compound of formula I having the structure:

wherein: W is selected from the group consisting of formula (i) or (ii) wherein the structures are as follows:

, , or in which K and E are independently selected from the group consisting of: N, CR and CZ, and wherein R, Z and Q are as defined below;

R₂ is selected from the group consisting of: H; a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃,

-SH; -SR₃; -SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN; -CO₂H; -

CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃; -NO₂; -SO₃H; -SOR₃; and -

SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z; R is selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃;

Q is selected from the group consisting of: NR₂; O; S; and C(R)₂; and X and Y are independently selected from the group consisting of: O; H; OH; and H_2; wherein the particles have a volume-weighted mean particle size of less than 5 μm.

34. The composition of claim 33, wherein volume-weighted mean particle size of less than 5 μm as measured by laser diffractometry.

35. The composition of claim 33 or 34, wherein Ri is H or CH₃.

36. The composition of any one of claims 33-35, wherein Q is NH.

37. The composition of any one of claims 33-36, wherein X and Y are oxygen.

38. The composition of any one of claims 33-37, wherein W is a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom; and wherein E, K and T, where present, are N or CH.

39. A method of producing a water soluble amorphous pharmaceutical composition, comprising combining a cyclodextrin and a compound of formula I having the structure:

wherein:

R₂ is selected from the group consisting of: H; a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃; -SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN; -CO₂H; - CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃; -NO₂; -SO₃H; -SOR₃; and - SO₂R₃, wherein R₃ is a linear, branched or cyclic, one to ten carbon saturated or unsaturated alkyl group; RCO-; ArCO-; and ArCH₂- wherein Ar is an aromatic substituent selected from the group consisting of: phenyl, napthyl, anthracyl, phenanthryl, furan, pyrrole, thiophene, benzofuran, benzothiophene, quinoline, isoquinoline, imidazole, thiazole, oxazole, and pyridine, and Ar may be optionally substituted with R or Z and R and Z; R is selected from the group consisting of: H; and, a structural fragment having a saturated or unsaturated linear, branched, or cyclic, skeleton containing one to ten carbon atoms, in which the carbon atoms may be optionally substituted with a substituent selected from the group consisting of: -OH; -OR₃; -O₂CR₃, -SH; -SR₃; -SOCR₃; -NH₂; -NHR₃; -NH(R₃)₂; -NHCOR₃; NRCOR₃; -I; -Br; -Cl; -F; -CN;

-CO₂H; -CO₂R₃; -CHO; -COR₃; -CONH₂; -CONHR₃; -CON(R₃)₂; -COSH; -COSR₃;

Z is an optional substituent selected from the group consisting of: H; -OH; -OR; -O₂CR; -SH; -SR; -SOCR; -NH₂; -NHR; -NH(R)₂; -NHCOR; NRCOR; -I; -Br;

-Cl; -F; -CN; -CO₂H; -CO₂R; -CHO; -COR; -CONH₂; -CONHR; -CON(R)₂; -COSH; -COSR; - NO₂; -SO₃H; -SOR; and, -SO₂R:

Q is selected from the group consisting of: NR₂; O; S; and C(R)₂; and

X and Y are independently selected from the group consisting of: O; H; OH; and H_2; wherein the particles have a volume-weighted mean particle size within the range of 0.1 μm and 50 μm.

40. The method of claim 39, wherein the volume-weighted mean particle size is measured by laser diffractometry.

41. The method of claim 39 or 40, wherein Ri is H or CH₃.

42. The method of any one of claims 39-41, wherein Q is NH.

43. The method of any one of claims 39-42, wherein X and Y are oxygen.

44. The method of any one of claims 39-43, wherein W is a five membered ring of formula (i) or (ii) comprising at least one nitrogen atom; and wherein E, K and T, where present, are N or CH.

45. The method of claim 39, wherein the pharmaceutical composition comprises a compound of the formula:

46. The method of claim 39, wherein the pharmaceutical composition comprises a compound of the formula:

47. The method of claim 39, wherein the pharmaceutical composition comprises a compound of the formula:

48. The method of claim 39, wherein the pharmaceutical composition comprises a compound of the formula:

49. The method of claim 39, wherein the pharmaceutical composition comprises a compound of the formula:

50. The method of any one of claims 39-49, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

51. The method of any one of claims 39-50, wherein the pharmaceutical composition is formulated for intravenous administration.

52. The method of any one of claims 39-51, wherein the particles have a volume- weighted mean particle size of less than 5 μm.

53. The method of claim 52, wherein the volume-weighted mean particle size is measured by laser diffractometry.

54. The method of any one of claims 39-53, wherein the cyclodextrin is selected from one or more of the following: a hydrophilic β-cyclodextrin derivative; a hydrophobic β- cyclodextrin; an ionaizable β-cyclodextrin derivative; an α-cyclodextrin; and a γ-cyclodextrin.

55. The method of any one of claims 39-53, wherein the cyclodextrin is selected from one or more of the following: methylated β-cyclodextrin; hydroxyalkylated β-cyclodextrin; branched β-cyclodextrin; alkylated β-cyclodextrin; acylated β-cyclodextrin; anionic β- cyclodextrin; an α-cyclodextrin; and a γ-cyclodextrin.

56. The method of any one of claims 39-55, wherein the cyclodextrin is 2- hydroxypropyl-β-cyclodextrin (HPCD).

57. The method of any one of claims 39-56, further comprising the addition of a water-soluble compound for inducing osmosis.

58. The method of claim 57, wherein the water-soluble compound for inducing osmosis is isotonic.

59. The method of claim 57 or 58, wherein the water-soluble compound for inducing osmosis is comprised of one or more of the following: sodium chloride, mannitol, dextrate, and dextrose.

60. The method of any one of claims 57-59, wherein the water-soluble compound for inducing osmosis is selected from: 0.9% sodium chloride and 5% dextrose.

61. The method of any one of claims 39-60, wherein a solution a solution is formed in an aqueous medium.

62. The method of any one of claims 39-61, wherein the method comprises stabilizing the water soluble amorphous pharmaceutical composition with one or more of: ultrasound energy; heat; elevated pressure; and mechanical agitation.

63. The method of claim 62, wherein stabilizing the water soluble amorphous pharmaceutical composition comprises the use of ultrasound energy.

64. The method of claim 62 or 63, wherein stabilizing the water soluble amorphous pharmaceutical composition comprises elevation of the water soluble amorphous pharmaceutical composition to a temperature of between about 4O⁰C and about 7O⁰C.

65. The method of any one of claims 62-64, wherein the mechanical agitation comprises stirring.