WO2006092017A1 - Combinaisons d'un glycoalcaloïde et d'un agoniste de tlr et diverses applications desdites combinaisons - Google Patents

Combinaisons d'un glycoalcaloïde et d'un agoniste de tlr et diverses applications desdites combinaisons Download PDF

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Publication number
WO2006092017A1
WO2006092017A1 PCT/AU2006/000269 AU2006000269W WO2006092017A1 WO 2006092017 A1 WO2006092017 A1 WO 2006092017A1 AU 2006000269 W AU2006000269 W AU 2006000269W WO 2006092017 A1 WO2006092017 A1 WO 2006092017A1
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group
glycoalkaloids
coramsine
compositions
radical
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PCT/AU2006/000269
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English (en)
Inventor
Stephen John Carter
Richard Lake
Dr Robbert Van Der Most
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Solbec Pharmaceuticals Limited
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Priority claimed from AU2005900969A external-priority patent/AU2005900969A0/en
Application filed by Solbec Pharmaceuticals Limited filed Critical Solbec Pharmaceuticals Limited
Priority to AU2006220238A priority Critical patent/AU2006220238A1/en
Priority to EP06704943A priority patent/EP1858526A1/fr
Publication of WO2006092017A1 publication Critical patent/WO2006092017A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/711—Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/12—Keratolytics, e.g. wart or anti-corn preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to the use of glycoalkaloids, such as solasonine and solamargine, in combination with TLR agonists to treat cancer and to compositions for use in such methods. More particularly the present invention relates to the use of glycoalkaloids in combination with TLR3, TLR7 or TLR9 agonists.
  • Cytotoxic chemotherapy remains one of the premier treatment options to combat cancer.
  • the efficacy of chemotherapy is limited by the fact that not all tumors respond optimally.
  • single-modality chemotherapy is rarely curative.
  • drug-resistant tumor cells often emerge when a single agent is used.
  • CORAMSINETM is mixture of solasonine and solamargine with anti-cancer properties.
  • TLRs Toll-Like receptors
  • PAMPs include various bacterial cell wall components such as Iipopolysaccharides (LPS), peptidoglycans and lipopeptides, as well as flagellin, bacterial DNA and viral double-stranded RNA. Stimulation of TLRs by PAMPs initiates a signalling cascade that involves a number of proteins, such as MyD88 and IRAK.
  • TLR2 is essential for the recognition of a variety of PAMPs, including bacterial lipoproteins, peptidoglycan, and lipoteichoic acids.
  • TLR3 is implicated in virus- derived double-stranded RNA.
  • TLR4 is predominantly activated by lipopolysaccharide.
  • TLR5 detects bacterial flagellin and TLR9 is required for response to unmethylated CpG DNA.
  • TLR7 and TLR8 have been shown to recognize small synthetic antiviral molecules.
  • the present invention seeks to improve the effectiveness and/or patient outcomes obtained using CORAMSINETM monotherapy through combination therapy with TLR agonists.
  • the present invention also provides a method of treating a tumorous growth comprising the step of administering a therapeutically effective amount of: (a) a first composition comprising at least two glycoalkaloids of formula I:
  • either one or both of the dotted lines represents a double bond, and the other a single bond, or both represent single bonds;
  • A represents a radical selected from the following radicals of general formulae (II) to (V):
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ; each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ; each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene; each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative; "X" is a radical selected from the group comprising -CH 2 -, -O- and -NH 2 -; and
  • the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof selected from the group comprising glyceric aldehyde, glycerose, erythrose, threose, ribose, arabinose, xylose, lyxose, altrose, allose, gulose, mannose, glucose, idose, galactose, talose, rhamnose, dihydroxyactone, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, and other hexoses, heptoses, octoses, nanoses, decoses, deoxysugars with branched chains, (e.g.
  • apiose hamamelose, streptose, cordycepose, mycarose and cladinose
  • compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar acids e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • benzimidazoles e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g
  • the present invention also provides a unit dosage form comprising a therapeutically effective amount of: (a) at least two glycoalkaloids of formula I: - A -
  • either one or both of the dotted lines represents a double bond, and the other a single bond, or both represent single bonds;
  • A represents a radical selected from the following radicals of general formulae (II) to (V):
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ;
  • each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ;
  • each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene;
  • each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative;
  • X is a radical selected from the group comprising -CH 2 -, -O- and -NH 2 -;
  • the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof selected from the group comprising glyceric aldehyde, glycerose, erythrose, threose, ribose, arabinose, xylose, lyxose, altrose, allose, gulose, mannose, glucose, idose, galactose, talose, rhamnose, dihydroxyactone, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, and other hexoses, heptoses, octoses, nanoses, decoses, deoxysugars with branched chains, (e.g.
  • apiose hamamelose, streptose, cordycepose, mycarose and cladinose
  • compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar acids e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • benzimidazoles e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g
  • the present invention still further provides a pack comprising a unit dosage form of glycoalkaloids and a unit dosage form of a TLR agonist.
  • Figure 1a is a graph illustrating the effect of different doses of intra-peritoneal CORAMSINETM on tumour size in mice;
  • Figure 1b is a graph plotting the survival of mice administered different doses of CORAMSINETM intra-peritoneally
  • Figure 2a is a graph illustrating the effect of different doses of intra-venous CORAMSINETM on tumour size in mice;
  • Figure 2b is a graph plotting the survival of mice administered different doses of CORAMSINETM intra-venously;
  • Figure 2c is a graph illustrating the effect of further (higher) doses of intra-venous CORAMSINETM on tumour size in mice;
  • Figure 2d is a graph plotting the survival of mice administered further (higher) doses of CORAMSINETM intra-venously
  • Figure 3a is a graph illustrating the effect of CORAMSINETM and FGK-45 monotherapy and combination therapy administered intra-peritoneally on tumour size in mice;
  • Figure 3b is a graph illustrating the survival of mice administered CORAMSINETM and FGK-45 monotherapy and combination therapy intra-peritoneally;
  • Figure 4a is a graph illustrating the effect of CORAMSINETM in combination with Poly I:C, loxoribine, CpG ODN or nonCpG ODN on tumour size in mice;
  • Figure 4b is a graph illustrating the survival of mice administered CORAMSINETM in combination with Poly I:C, loxoribine, CpG ODN or nonCpG ODN
  • Figure 4c is a graph illustrating the effect of CORAMSINETM in combination with CpG ODN or nonCpG ODN on tumour size in mice;
  • Figure 4d is a graph illustrating the survival of mice administered CORAMSINETM in combination with CpG ODN or nonCpG ODN;
  • Figure 5 is a plot illustrating the results of ApoStat staining in cells exposed to CORAMSINETM or gemcitabine;
  • Figure 6a is a graph illustrating the IL-6 levels when cells are exposed to CORAMSINETM.
  • Figure 6b is a graph illustrating the IL-6 levels when cells are exposed to gemcitabine.
  • the present invention also provides a method of treating a tumorous growth comprising the step of administering a therapeutically effective amount of: (a) a first composition comprising at least two glycoalkaloids of formula I:
  • either one or both of the dotted lines represents a double bond, and the other a single bond, or both represent single bonds;
  • A represents a radical selected from the following radicals of general formulae (II) to (V):
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ; each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ; each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene; each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative; "X" is a radical selected from the group comprising -CH 2 -, -O- and -NH 2 -; and
  • the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof selected from the group comprising glyceric aldehyde, glycerose, erythrose, threose, ribose, arabinose, xylose, lyxose, altrose, allose, gulose, mannose, glucose, idose, galactose, talose, rhamnose, dihydroxyactone, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, and other hexoses, heptoses, octoses, nanoses, decoses, deoxysugars with branched chains, (e.g.
  • apiose hamamelose, streptose, cordycepose, mycarose and cladinose
  • compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar acids e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • benzimidazoles e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g
  • the present invention is based on the surprising discovery that glycoalkaloids can cause cell death without stimulating the immune system.
  • glycoalkaloids can cause cell death without stimulating the immune system.
  • the glycoalkaloids and the TLR agonist are complimentary and/or synergistic in that they provide a better patient outcome relative to the respective monotherapies and relative to the additive effects of the respective monotherapies.
  • the present invention also provides a method for treating a tumorous growth comprising the steps of administering a therapeutically effective synergistic amount of the first and second compositions mentioned above.
  • the tumorous growth may be associated with a range of cancers including cancer selected from the group consisting of: melanomas and non-melanoma skin including lignin melanoma, solar keratosis, basal cell carcinoma, squamous cell carcinoma and actinic keratoses; mesothelioma; breast; prostate; liver; lung; colon; rectum; urinary bladder; non-Hodgkin lymphoma; kidney; pelvis; pancreas; pharynx; head & neck; ovarian; oral; thyroid; stomach; brain; multiple myeloma; oesophagus; liver and intrahepatic bile duct; cervix; larynx; acute myeloid leukemia; chronic lymphocytic leukemia; heart; Hodgkin lymphoma; testis; small intestine; chronic myeloid leukemia; acute lymphocytic leukemia; gallbladder; bones and joints; eye; nose;
  • glycoalkaloids used in the method of the present invention may be varied.
  • the glycoalkaloids are triglycoside glycoalkaloids, solasodine glycosides or are selected from the group of glycoalkaloids consisting of: solamargine, solasonine, solanine, tomatine, solanocapsine and 26- aminofurostane.
  • the glycoalkaloids may be chiral, stereoisomers and mixtures thereof including enantiomers and/or diastereoisomers. Furthermore, the glycoalkaloids may be obtained from natural sources, synthesized or produced by chemically modifying other glycoalkaloids.
  • the number of glycoalkaloids used may be varied, as may their relative ratios in the composition. However, when the composition comprises two glycoalkaloids they may be present in a ratio selected from the group of ratios consisting of approximately: 1 :6 - 1 :0.5; 1 :5; 1 :4; 1 :3; 1 :2, 1 : 1.5 and 1 : 1.
  • the glycoalkaloids are solamargine and solasonine in a ratio between about 1 :6 and 6:1 or more preferably in a ratio between about 1 :4 and 4:1 , 1 :3 and 3:1 or 1 :2 to 2:1.
  • the glycoalkaloids are solamargine and solasonine and they are present in a 1 :1 ratio it is preferred that the glycoalkaloids are isolated.
  • the glycoalkaloids are solasonine and solamargine it is preferred that they do not constitute 66% of glycosides in the composition.
  • the glycoalkaloids constitute greater than 70%-90% of the glycosides in the composition, more preferably 91-95% and even more preferably 96-100% of the glycosides in the composition.
  • compositions of the present invention may be varied depending on their intended end use.
  • the compositions comprise about 0.001% - 5% or 10% glycoalkaloids, more preferably 0.01 % - 5% or 10% and even more preferably 0.1%- 5% or 10% glycoalkaloids.
  • the actual concentration of glycoalkaloids in the composition will vary and depend at least on the nature of the tumour being treated and the condition of the subject to be treated. Skilled practitioners can determine the most appropriate dose using their ordinary skill and taking into account various parameters that apply in such situations. For example, the higher the cancer load in a particular patient the higher the dose of glycoalkaloids that can be administered and well tolerated by the patient.
  • the concentration of glycoalkaloids administered as part of the combination therapy is less than in a comparable situation in which it was to be administered as a monotherapy.
  • the amount of glycoalkaloid may be about 0.1mg/kg - 100mg/kg, 1mg/kg-80mg/kg, 5mg/kg-60mg/kg or 10mg/kg-40mg/kg.
  • the amount of glycoalkaloid is about 0.5-5mg/kg, 0.75-4mg/kg or 1- 3mg/kg.
  • the TLR agonist is a TLR3, TLR7, TLR8 or TLR9 agonist.
  • the TLR3 agonist may be double-stranded RNA (dsRNA) containing a molecular pattern associated with viral infection. More particularly the TLR3 agonist may be polyinosine-polycytidylic acid (poly(l:C)), a synthetic analog of dsRNA.
  • dsRNA double-stranded RNA
  • poly(l:C) polyinosine-polycytidylic acid
  • TLR7/TLR8 agonists include imidazoquinoline compounds such as ImiquimodTM (compound R387), guanine analogs such as loxoribine, single stranded PoIyU, single stranded RNA40 and R484, a small anti-viral molecule.
  • the agonist when it is a TLR9 agonist it may be a nucleotide comprising a sequence corresponding to a pathogen associated microbial pattern (PAMPs). More particularly, the TLR9 agonist is a CpG ODN such as a nucleotide comprising a sequence according to formula I:
  • C and G are unmethylated, X-i, X 2 , X 3 and X 4 are nucleotides and a GCG trinucleotide sequence is not present at or near the 5' and 3' termini.
  • Xi X 2 may be the dinucleotide GpA and/or X 3 X 4 may be the dinucleotide TpC or TpT.
  • Xi X 2 CGX 3 X 4 is preceded on the 5 1 end by a T.
  • the nucleotide comprises the sequence TGACGTT or TGACGTC.
  • the nucleotide may be of various sizes but preferably the nucleotide is about 8 to 40 bases or about 16-20 bases. In one particular example the nucleotide is 5' TCC ATG ACG TTC CTG ATG CT 3' (CpG 1668).
  • TLR9 agonists include oligonucleotides with human specific type B CpGs (e.g. ODN 2006), oligonucleotides with human specific type A CpGs (e.g. ODN 2216) and oligonucleotides with human specific type C CpGs (e.g. ODN M362).
  • Stabilized oligonucleotides include those with a modified phosphate backbone e.g. a phosphorothioate modified phosphate backbone (i.e. at least one of the phosphate oxygen's is replaced by sulfur).
  • Other stabilized oligonucleotides include: nonionic DNA analogs, such as alkyl- and aryl- phosphonates, phosphodiester and alkylphosphotriesters.
  • Oligonucleotides which contain a diol, such as tetraethyleneglycol or hexaethyleneglycol, at either or both termini have are also known to be resistant to nuclease degradation.
  • the concentration of TLR agonist in the composition will vary and depend at least on the nature of the tumour being treated and the condition of the subject to be treated. Skilled practitioners can determine the most appropriate dose using their ordinary skill and taking into account various parameters that apply in such situations.
  • the concentration of agonist administered as part of the combination therapy is less than in a comparable situation in which it was to be administered as a monotherapy.
  • the amount of agonist may be about 0.1mg/kg - 100mg/kg, 1mg/kg-80mg/kg, 5mg/kg-60mg/kg or 10mg/kg-40mg/kg.
  • the amount of agonist is 0.005mg/kg-10mg/kg and more preferably 0.01-1 mg/kg.
  • the present invention also provides a method of treating a tumorous growth comprising the step of administering a therapeutically effective amount of: (a) a first composition comprising at least two glycoalkaloids of formula I:
  • either one or both of the dotted lines represents a double bond, and the other a single bond, or both represent single bonds;
  • A represents a radical selected from the following radicals of general formulae (II) to (V):
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ; each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ; each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene; each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative; "X" is a radical selected from the group comprising -CH 2 -, -O- and -IMH2-; and
  • the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof selected from the group comprising glyceric aldehyde, glycerose, erythrose, threose, ribose, arabinose, xylose, lyxose, altrose, allose, gulose, mannose, glucose, idose, galactose, talose, rhamnose, dihydroxyactone, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, and other hexoses, heptoses, octoses, nanoses, decoses, deoxysugars with branched chains, (e.g.
  • apiose hamamelose, streptose, cordycepose, mycarose and cladinose
  • compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar acids e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • benzimidazoles e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g
  • the third composition is preferably an anti-CD40 antibody such as FGK-45.
  • FGK-45 an anti-CD40 antibody
  • a combination including FGK-45 was found to have very potent anti-tumour activity (results not shown).
  • compositions can be formulated in various ways depending on the route and mode of administration and whether the compositions are combined in a single dosage unit or are administered as separate dosage units.
  • the compositions used in the method of the invention may further comprise a pharmaceutically acceptable carrier.
  • compositions comprising one or more active ingredients are generally known in the art.
  • Such pharmaceutical compositions will generally be formulated for the mode of delivery that is to be used and will usually include one or more pharmaceutically acceptable carriers.
  • compositions of the invention may further comprise suitable carriers, excipient and diluents that are pharmaceutically acceptable and compatible with the active ingredient.
  • Suitable carriers, excipient and diluents include, without limitation, water, saline, ethanol, dextrose, cyclodextrins such as hydroxy propyl beta-cyclodextrin, glycerol, lactose, dextrose, sucrose sorbitol, mannitol, starches, gum acacia, calcium phosphates, alginate, tragacanth, gelatine, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methyl cellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate and mineral oil or combinations thereof.
  • suitable carriers, excipient and diluents include, without limitation, water, saline, ethanol, dextrose, cyclodextrins such as hydroxy propyl beta-cyclodextrin, glycerol, lactose, dextrose, sucrose sorbi
  • compositions can additionally include lubricating agents, pH buffering agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavouring agents.
  • lubricating agents pH buffering agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavouring agents.
  • the particular selection of constituent that can be included in the compositions described herein will generally depend on the particular mode of delivery used to bring the active agents into contact with their target cells or tissue.
  • the first and second compositions are administered in separate dosage forms.
  • the compositions may be administered simultaneously or sequentially.
  • “simultaneously” means that the compositions are administered at the same time or within one hour of each other.
  • the compositions are administered greater than one hour apart they are deemed to be administered sequentially.
  • compositions of the present invention should be administered in dosage unit form that is therapeutically effective.
  • Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated, each unit containing a quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the dosage unit forms of the invention are dictated by and directly dependent on at least (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved and (b) the limitations inherent in the art of compounding such an active material for the treatment sought.
  • the quantity of active compound to be administered will be largely dependent on the toxicity and specific activity of compound, the subject to be treated and the degree of treatment required. Precise amounts of compound required to be administered may depend on the judgement of the practitioner and may be peculiar to each subject.
  • compositions may be given as a single dose schedule, or more preferably, in a multiple dose schedule.
  • a multiple dose schedule is one in which a primary course of delivery may be with 1 to 100 separate doses, followed by other doses given at subsequent time intervals required to maintain or reinforce the treatment.
  • the dosage regimen will also, at least in part, be determined by the need of the individual and the judgement of the practitioner.
  • compositions are administered and their timing can be varied and will be determined by a practitioner of ordinary skill on a case by case basis.
  • the compositions be administered as separate dosage forms
  • the first and second compositions can be formulated into a single dosage form for more convenient administration.
  • unwanted interactions between the compositions must be managed to ensure the therapeutic activity of the compositions is not unduly compromised.
  • compositions may be administered by any route deemed appropriate by the practitioner based on the particular circumstances of a given case. These routes include: intradermal, transdermal, intramuscular, intraperitoneal, intravenous, subcutaneous, oral, topical, epidural, buccal, rectal, vaginal and intranasal.
  • compositions according to the invention may be administered to a patient using any technology or delivery route that permits contact between the active agents and their target site.
  • the composition is via the intraperitoneal or intra venous route.
  • any technology that allows targeted delivery of the pharmaceutical composition via, subcutaneously, intramuscularly, intraorbitally, ophthalmically, intraventricularly, intracranially, intracapsularly, intraspinally, intracistemally, buccal, rectally, vaginally, intranasally or by pulmonary administration may be used to deliver the composition.
  • the mode of administration must, however, be at least suitable for the form in which the composition has been prepared.
  • the mode of administration for the most effective response may need to be determined empirically and the means of administration described below are given as examples and do not limit the method of delivery of the composition of the present invention in any way. All the formulations described below are commonly used in the pharmaceutical industry and are commonly known to suitably qualified practitioners.
  • compositions may be adapted for parenteral administration that facilitates delivery of a therapeutically effective amount of the glycoalkaloids and TLR agonist to the target cell or tissue.
  • compositions suitable for injectable use include sterile aqueous solutions (where water-soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the glycoalkaloids may be encapsulated in liposomes and delivered in injectable solutions to assist their transport across cell membrane.
  • the solution may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol and liquid polyethylene glycol and the like), suitable mixtures thereof and vegetable oils or cyclodextrins such as hydroxy propyl beta-cyclodextrin.
  • Proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of superfactants.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatine.
  • Sterile injectable solutions may be prepared by incorporating the glycoalkaloids in the required amount in an appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilisation.
  • dispersions are prepared by incorporating the various sterilised active ingredient into a sterile vehicle that contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum drying and freeze-drying techniques that yield a powder of the active ingredient plus, optionally, any additional desired ingredient from previously sterile-filtered solution thereof.
  • Topical delivery systems may be appropriate for administering the compositions of the present invention, particularly the glycoalkaloids, depending upon the preferred treatment regimen.
  • Topical formulations may be produced by dissolving or combining the active agent in an aqueous or nonaqueous carrier.
  • any liquid, cream, or gel, or similar substance that does not appreciably react with the active or any other of the ingredients that may be introduced into the composition and which is non-irritating is suitable.
  • Appropriate non-sprayable viscous, semi-solid or solid forms can also be employed that include a carrier compatible with topical application and have a dynamic viscosity preferably greater than water.
  • Suitable formulations are well known to those skilled in the art and include, but are not limited to, solutions, suspensions, emulsions, creams, gels, ointments, powders, liniments, salves, aerosols, transdermal patches, etc, which are, if desired, sterilized or mixed with auxiliary agents, e.g., preservatives, stabilizers, emulsifiers, wetting agents, fragrances, colouring agents, odour controllers, thickeners such as natural gums etc.
  • Particularly preferred topical formulations include ointments, creams or gels.
  • Ointments generally are prepared using either (1) an oleaginous base, i.e., one consisting of fixed oils or hydrocarbons, such as white petroleum or mineral oil, or (2) an absorbent base, i.e., one consisting of an anhydrous substance or substances which can absorb water, for example anhydrous lanolin.
  • an oleaginous base i.e., one consisting of fixed oils or hydrocarbons, such as white petroleum or mineral oil
  • an absorbent base i.e., one consisting of an anhydrous substance or substances which can absorb water, for example anhydrous lanolin.
  • the glycoalkaloid is added to an amount affording the desired concentration.
  • Creams are oil/water emulsions. They consist of an oil phase (internal phase), comprising typically fixed oils, hydrocarbons and the like, waxes, petroleum, mineral oil and the like and an aqueous phase (continuous phase), comprising water and any water-soluble substances, such as added salts.
  • the two phases are stabilised by use of an emulsifying agent, for example, a surface active agent, such as sodium lauryl sulfate; hydrophilic colloids, such as acacia colloidal clays, veegum and the like.
  • an emulsifying agent for example, a surface active agent, such as sodium lauryl sulfate; hydrophilic colloids, such as acacia colloidal clays, veegum and the like.
  • Gels comprise a base selected from an oleaginous base, water, or an emulsion- suspension base.
  • a gelling agent that forms a matrix in the base, increasing its viscosity.
  • examples of gelling agents are hydroxypropyl cellulose, acrylic acid polymers and the like.
  • the active is added to the formulation at the desired concentration at a point preceding addition of the gelling agent.
  • the amount of the glycoalkaloid compound incorporated into a topical formulation is not critical; the concentration should be within a range sufficient to permit ready application of the formulation to the affected tissue area in an amount that will deliver the desired amount of glycoalkaloid to the desired treatment site.
  • compositions adapted for oral administration in such a manner that facilitates delivery of a therapeutically effective concentration of the active agent(s) to the target cell or tissue.
  • the effective dosages of the glycoalkaloid when administered orally, must take into consideration the diluent, preferably water.
  • the composition preferably contains about 1 to about 200mg active agent such as glycoalkaloids.
  • active agent such as glycoalkaloids.
  • Solid dosage forms are described generally in Martin, Remington's Pharmaceutical Sciences, 18th Ed. (1990 Mack Publishing Co. Easton PA 18042) at Chapter 89.
  • Solid dosage forms include tablets, capsules, pills, troches or lozenges, cachets or pellets.
  • liposomal or proteinoid encapsulation may be used to formulate compositions (as, for example, proteinoid microspheres reported in U.S. Patent No. 4,925,673).
  • Liposomal encapsulation may be used and the liposomes may be derivatised with various polymers (e.g., U.S. Patent No.
  • composition will also include inert ingredients that allow for protection against the stomach environment and release of the active agents in inappropriate areas of the body.
  • the active agent(s) in the stomach may be desirable to release the active agent(s) in the stomach, the small intestine (the duodenum, the jejunem, or the ileum), or the large intestine.
  • the small intestine the duodenum, the jejunem, or the ileum
  • the large intestine One skilled in the art has available formulations that will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine.
  • the release will avoid the deleterious effects of the stomach environment, either by protection of the composition or by release of the glycoalkaloids or TLR agonist beyond the stomach environment, such as in the intestine.
  • a coating impermeable to at least pH 5.0 is essential.
  • examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S and Shellac. These coatings may be used as mixed films.
  • a coating or mixture of coatings that are not intended for protection against the stomach can also be used on tablets. This can include sugar coatings, or coatings that make the tablet easier to swallow.
  • Capsules may consist of a hard shell (such as gelatine) for delivery of dry therapeutic i.e. powder; for liquid forms, a soft gelatine shell may be used.
  • the shell material of cachets could be thick starch or other edible paper. For pills, lozenges, moulded tablets or tablet triturates, moist massing techniques can be used.
  • compositions may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavouring agents.
  • diluents could include carbohydrates, especially mannitol, alpha-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch.
  • Certain inorganic salts may be also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride.
  • Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.
  • Disintegrants may be included in the pharmaceutical formulations of the present invention.
  • Materials used as disintegrants include but are not limited to starch including the commercial disintegrant based on starch, Explotab.
  • Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramyiopectin, sodium alginate, gelatine, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used.
  • Other disintegrants include insoluble cationic exchange resins.
  • Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.
  • Binders may be used to hold the pharmaceutical composition together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch and gelatine. Others include methylcellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic.
  • MC methylcellulose
  • EC ethyl cellulose
  • CMC carboxymethyl cellulose
  • PVP polyvinyl pyrrolidone
  • HPMC hydroxypropylmethyl cellulose
  • Lubricants may be used as a layer between the glycoalkaloid and the die wall and these can include but are not limited to: stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium Iauryl sulfate, polyethylene glycol of various molecular weights and Carbowax 4000 and 6000.
  • the glidants may include starch, talc, pyrogenic silica and hydrated silicoaluminate.
  • a surfactant might be added as a wetting agent.
  • Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate.
  • anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate.
  • Cationic detergents might be used and could include benzalkonium chloride or benzethomium chloride.
  • nonionic detergents that could be included in the pharmaceutical composition as surfactants are lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the composition either alone or as a mixture in different ratios.
  • Additives which potentially enhance uptake of the glycoalkaloids, are for instance the fatty acids oleic acid, linoleic acid and linolenic acid.
  • Controlled release formulations may be desirable.
  • the compositions could be incorporated into an inert matrix that permits release by either diffusion or leaching mechanisms i.e., gums.
  • Slowly degenerating matrices may also be incorporated into the pharmaceutical composition.
  • Another form of a controlled release is by a method based on the Oros therapeutic system (Alza Corp.), i.e. the composition is enclosed in a semipermeable membrane which allows water to enter and push the composition out through a single small opening due to osmotic effects. Some enteric coatings also have a delayed release effect.
  • Film coating may be carried out in a pan coater or in a fluidised bed or by compression coating.
  • the active agents may be included in the pharmaceutical composition as fine multiparticulates in the form of granules or pellets of particle size about 1mm.
  • the formulation for capsule administration could also be as a powder, lightly compressed plugs or even as tablets.
  • the active agent(s) could be prepared by compression.
  • Microparticles may be made by a variety of methods known to those in the art, for example, solvent evaporation, desolvation, complex coacervation, polymer/polymer incompatibility, interfacial polymerisation etc.
  • Hydrophilic polymers forming the microparticles may be attached to a targeting protein that acts to enable the microparticle to specifically bind selected target cells or tissues bearing the target molecule (e.g. characteristic marker).
  • the hydrophilic polymers may be conjugated to the Fab 1 fragment of an antibody. Smaller peptides from the hypervariable region or from another peptide interacting with a specific cell surface ligand may also be conjugated to the complexes. It is most preferred that the antibodies or antibody fragments are directed against target molecules associated with cancerous tissues or cells.
  • the targeting protein e.g. an antibody or an antibody fragment
  • the targeting protein can be attached to the hydrophilic polymers either before or after formation of the microparticle.
  • the targeting protein is coupled to the hydrophilic polymer, where the targeting protein/hydrophilic polymer is subsequently used to form the microparticle complex.
  • This provides a convenient means for modifying the targeting specificity of an otherwise generic microparticle.
  • Targeted microparticles may be prepared by incorporating the Fab 1 fragment into the microparticles by a variety of techniques well known to those of skill in the art. For example, a biotin conjugated Fab' may be bound to a microparticle containing a streptavidin.
  • the biotinylated Fab' may be conjugated to a biotin derivatised microparticle by an avidin or streptavidin linker. Typically about 30 to 125 and more typically about 50 to 100 Fab' fragments per microparticle complex are used.
  • compositions of the present invention may also be formed into powders or some other form that is suitable for delivery by inhalation. Whilst inhalation may be via the mouth it will be appreciated that the route of delivery may also by via the nose.
  • compositions are particularly useful for treatment of diseases or disorders of the respiratory system such as lung cancer or cancer that may affect other parts of the respiratory system.
  • compositions for delivery to the lungs they are preferably designed to reach the site of the alveoli.
  • the compositions may contain various doses of active agent and the particular dose will be determined by a skilled practitioner with due consideration to the recipient and the state of the disease to be treated.
  • the compositions contain between about 100ug- 100mg of glycolalkaloids, about 200ug-50mg of glycoalkaloids or 200ug - 10mg glycoalkaloids.
  • the method of the present invention involves the administration of glycoalkaloids and TLR agonists.
  • Particular dosage forms may be produced that are particularly useful and these form separate aspects of the present invention.
  • the present invention also provides a unit dosage form comprising a therapeutically effective amount of: (a) at least two glycoalkaloids of formula I:
  • either one or both of the clotted lines represents a double bond, and the other a single bond, or both represent single bonds;
  • A represents a radical selected from the following radicals of general formulae (II) to (V):
  • each of R 1 is a radical separately selected from the group consisting of hydrogen, amino, oxo and OR 4 ;
  • each of R 2 is a radical separately selected from the group consisting of hydrogen, amino and OR 4 ;
  • each of R 3 is a radical separately selected from the group consisting of hydrogen, alkyl and R 4 -alkylene;
  • each of R 4 is a radical separately selected from the group consisting of hydrogen, carbohydrate and a carbohydrate derivative;
  • X is a radical selected from the group comprising -CH 2 -, -O- and -NH 2 -;
  • the compound includes at least one R 4 group in which R 4 is a carbohydrate or a derivative thereof selected from the group comprising glyceric aldehyde, glycerose, erythrose, threose, ribose, arabinose, xylose, lyxose, altrose, allose, gulose, mannose, glucose, idose, galactose, talose, rhamnose, dihydroxyactone, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, and other hexoses, heptoses, octoses, nanoses, decoses, deoxysugars with branched chains, (e.g.
  • apiose hamamelose, streptose, cordycepose, mycarose and cladinose
  • compounds wherein the aldehyde, ketone or hydroxyl groups have been substituted e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar acids e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • benzimidazoles e.g. N-acetyl, acetyl, methyl, replacement of CH 2 OH
  • sugar alcohols e.g
  • the active components may be provided as separate unit dosage forms in a pack or kit arrangement.
  • the present invention also provides a pack comprising a unit dosage form of glycoalkaloids and a unit dosage form of a TLR agonist.
  • derived and "derived from” shall be taken to indicate that a specific integer may be obtained from a particular source albeit not necessarily directly from that source.
  • the term "isolated” means essentially free of (i) mono and diglycosides and, preferably, essentially free of (ii) free sugars such as mono, di, tri, oligo or polysaccharides and (iii) aglycone.
  • steps are taken to stabilise the glycoalkaloids, it will be appreciated that even in an isolated glycoalkaloid composition of the present invention there will be a small amount of free sugars and mono and diglycosides that result from degradation of the glycoalkaloids.
  • composition consisting essentially of means that the glycoalkaloids in the composition are the only glycosides therein.
  • a composition consisting essentially of solamargine and solasonine includes solamargine and solasonine and may include other non-glycoside constituents.
  • the murine AB1-HA (1 , 2) and human JU77 and LO68 (3) mesothelioma cell lines were cultured in DMEM (Invitrogen) supplemented with 20 mM HEPES, 0.05 mM 2-mercaptoethanol, 100 units/ml penicillin (CSL, Melbourne, Australia), 50 ⁇ g/ml gentamicin (David Bull Labs, Warwick, UK) and 5% FCS (Invitrogen).
  • SBP002 (CORAMSINETM) - a 1 :1 solamargine and solasonine formulation in 3% acetic acid was dissolved to 200 ⁇ g/ml in 3% filter sterilised glacial acetic acid.
  • mice BALB/c (H-2 d ) mice were obtained from the Animal Resources Centre (Perth, WA, Australia) and maintained under standard conditions.
  • AB1-HA cells (1 x 10 6 ) were inoculated subcutaneously (s.c.) on one side of the ventral surface in the lower flank region.
  • treatment commenced 10 days later when a small palpable lump was evident, ranging from 1 to 2 mm in diameter (day 0).
  • CORAMSINETM was injected into mice either intraperitoneal (i.p.) or intravenous (i.v.) in a range of doses and volumes to optimize the treatment regimen: i.v.
  • mice were given in the tail vein of mice in 500 - 1000 ⁇ l volume (dose diluted in saline) and i.p. doses were given in a total of 200 ⁇ l.
  • Acetic acid diluted in saline was used as the control treatment in the majority of experiments. This control was selected as CORAMSINETM is initially resuspended in 3% glacial acetic acid before diluting to treatment concentrations. Tumor size was measured with callipers three times weekly during the course of chemotherapy and subsequently until tumor size reached 100 mm 2 at which point mice were euthanized.
  • mice receiving activating anti-CD40 antibody received 100 ⁇ g in 100 ⁇ l PBS i.v. three times in 6 days (days 4, 6, and 9).
  • mice with palpable tumours were injected i.t. with 10 ⁇ g polyinosinic:cytidylic acid
  • Caspase induction was detected through the use of ApoStat-FITC (R&D, MN, US). 2 x 10 5 AB1-HA cells (logarithmic growth) were inoculated into individual wells of 6-well plates and incubated overnight at 37 0 C with 5% CO 2 . Individual wells were treated with dilutions of gemcitabine or CORAMSINETM (in a total of 1 ml media). 30 minutes prior to completion of the incubation, 10 ⁇ l of 50 ⁇ g/ml ApoStat was added. After 30 minute incubation with ApoStat, cells were harvested, washed in PBS and resuspended in 400 ⁇ l of PBS.
  • mice were injected with 1 x 10 6 AB1-HA cells subcutaneously in the flank and assessed every 2 days for tumour growth until the tumour became palpable (day 0).
  • CORAMSINETM 14 mg/kg or saline/acetic acid solution equivalent to the 14 mg/kg dose (as control) was injected i.p. in a 300 ⁇ l volume at days 0, 1 , 2, 7, 8, 9, 14, 15 and 16. Mice were sacrificed at Day 3, 10 and 17. Tumors were excised and made into histological blocks. Sections of the tumors were stained with TUNEL (Roche, Castle Hill, NSW, Australia) as per manufacturers protocol.
  • the IL-6 assay was performed using a human IL-6 ELISA kit (eBioscience, USA) according to the manufacturers protocol. Briefly, 96-well plates were incubated at room temperature overnight with 100 ⁇ l of anti-human IL-6 monoclonal antibody. After washing and blocking steps, 100 ⁇ l of each experimental sample was added, followed by incubation with streptavidin-HRP conjugate and ABTS substrate. Plates were read at 405 nm on a SpectraMax 250 plate reader (Molecular Devices) and the concentration of IL-6 for each sample was calculated from the standard curve. Samples for analysis in the ELISA were prepared by seeding cells in 96-well plates with or without CORAMSINETM or gemcitabine. Chemotherapeutic agents were added 24 hours after cell seeding. The concentration of IL-6 shown represents the mean value generated from three separate experiments.
  • AB1-HA mesothelioma cells (10 6 ) were inoculated subcutaneously and treatment was initiated when tumors became palpable (usually 10 days after inoculation).
  • Chemotherapy treatment protocols were optimised by varying the route of administration, the dose, and the treatment schedule.
  • mice were injected with different doses of CORAMSINETM (12 and 14 mg/kg) once AB1-HA tumors were palpable.
  • a treatment schedule of 'three days on - four days off' was optimal in the context of i.p. injection (i.e., mice did not tolerate daily injections for more than three days). Therefore, we used this protocol throughout this study.
  • This dosing schedule significantly slowed tumor growth during the first two weeks of treatment with 12 or 14 mg/kg CORAMSINETM (Fig.ia). In fact, tumor size even decreased at day 15 after treatment with 14 mg/ml CORAMSINETM. However, mice did not tolerate further treatment beyond day 15, and tumor growth rates increased after treatment stopped. Nevertheless, i.p. CORAMSINETM treatments at these doses did prolong survival time, although all mice eventually succumbed to the tumor (Fig.1 b).
  • mice (5/group) were inoculated s.c. with AB1-HA and treated with a single high dose of CORAMSINETM (30 mg/kg) when their tumors had reached 77 ⁇ 23 mm 2 .
  • Example 2 - CORAMSINETM is potentiated by immunotherapy with CpG- containing oligodeoxynucleotides.
  • Example 3 - CORAMSINETM does not induce cell death through apoptosis.
  • CORAMSINETM-exposed cells do take up Pl (see the increased number of PI + /ApoStat " cells in CORAMSINETM-treated cells), which is indicative of primary necrosis when observed in the absence of indications for apoptosis (32,33).
  • gemcitabine treatment (1-10 ⁇ g/ml) did induce apoptosis using ApoStat staining (Fig.5) .
  • TUNEL staining revealed similar numbers of apoptotic cells in CORAMSINETM- (10 ⁇ g/ml, 16 hours) and PBS- treated AB1 cells (data not shown).
  • CORAMSINETM led to a decrease of IL-6 production as cell viability decreased

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Abstract

La présente invention décrit une méthode de traitement d'une tumeur en croissance qui comprend une étape d'administration d'une quantité thérapeutiquement significative de : (a) une première composition comprenant au moins deux glycoalcaloïdes, et (b) une seconde composition comprenant un agoniste de TLR.
PCT/AU2006/000269 2005-03-02 2006-03-01 Combinaisons d'un glycoalcaloïde et d'un agoniste de tlr et diverses applications desdites combinaisons WO2006092017A1 (fr)

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WO2008064425A1 (fr) * 2006-11-30 2008-06-05 Solbec Pharmaceuticals Limited Combinaisons de glycoalkaloïdes et d'agents chimiothérapeutiques et utilisations diverses
EP2116602A1 (fr) * 2008-05-07 2009-11-11 Institut Gustave Roussy Combinaison de produits pour traiter le cancer
US8557247B2 (en) 2007-05-24 2013-10-15 Glaxosmithkline Biologicals Sa Lyophilised antigen composition

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WO2004002497A1 (fr) * 2002-07-01 2004-01-08 Solbec Pharmaceuticals Limited Compositions contenant des glyco-alcaloides et leurs diverses utilisations

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WO2000061153A1 (fr) * 1999-04-09 2000-10-19 Cura Nominees Pty. Ltd. Compositions medicinales et leur procede de preparation
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008064425A1 (fr) * 2006-11-30 2008-06-05 Solbec Pharmaceuticals Limited Combinaisons de glycoalkaloïdes et d'agents chimiothérapeutiques et utilisations diverses
US8557247B2 (en) 2007-05-24 2013-10-15 Glaxosmithkline Biologicals Sa Lyophilised antigen composition
EP2116602A1 (fr) * 2008-05-07 2009-11-11 Institut Gustave Roussy Combinaison de produits pour traiter le cancer
WO2009136282A1 (fr) * 2008-05-07 2009-11-12 Institut Gustave Roussy Associations médicamenteuses pour traiter le cancer

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