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WO2013004658A1 - Formulations de 5-fluorocytosine et leurs utilisations - Google Patents

Formulations de 5-fluorocytosine et leurs utilisations Download PDF

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Publication number
WO2013004658A1
WO2013004658A1 PCT/EP2012/062822 EP2012062822W WO2013004658A1 WO 2013004658 A1 WO2013004658 A1 WO 2013004658A1 EP 2012062822 W EP2012062822 W EP 2012062822W WO 2013004658 A1 WO2013004658 A1 WO 2013004658A1
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Prior art keywords
batch
granule formulation
granule
formulation
dissolution
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Inventor
Jérôme HECQ
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Transgene SA
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Transgene SA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5161—Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine

Definitions

  • the invention relates to formulations of 5-fluorocytosine, and to use of them for treating hy erproliferative diseases such as cancer and other disease, such as fungal diseases.
  • the invention further relates to the manufacture of said formulations.
  • TK thymidine kinase
  • GCV ganciclovir
  • CDase cytosine deaminase
  • 5-FC 5-fluorocytosine
  • CDase is involved in the pyrimidine metabolic pathway by which exogenous cytosine is transformed into uracil by means of a hydrolytic deamination. While CDase activities have been demonstrated in prokaryotes and lower eukaryotes, especially some fungal cells, they are not present in mammals. CDase is further able to deaminate an analogue of cytosine, the 5- fluorocytosine (5-FC), thereby forming 5-fluorouracil (5-FU), which is further converted to metabolites some of which (e.g. 5-fluoro-UMP (5-FUMP)) interfere with fungal RNA and DNA synthesis leading to strong cytotoxicity and cell death.
  • 5-fluoro-UMP 5-fluoro-UMP
  • 5-fluorocytosine is a synthetic drug marketed by Valeant Pharmaceuticals in various countries under brand names such as Ancotil (Registered trademark) and Ancobon (Registered trademark). Both of these products are immediate release tablet and capsule formulation, respectively.
  • 5-FC is well absorbed after oral administration, and penetrates into the body tissues well. This drug is used for treating severe systemic mycoses, such as cryptococcosis, candidosis, chromoblastomycosis and aspergillosis.
  • 5-FC has been successfully used combined with vector based delivery and expression of exogenous CDase activity into cancer cells (see Erbs et al., 2008, Cancer Gene Therapy, 15, 18-28; Dias et al, Clin Cancer Res (2010),16,2540-9; Foloppe et al., 2008, Gene Ther., 15, 1361-1371).
  • This approach to fight cancer consists in local chemotherapy directed by a suicide gene. More particularly, a viral-based vector carrying a suicide gene, e.g. the gene FCUl (see W099/54481), is injected directly into the tumor before 5-FC administration. This FCUl gene is encoding CDase activity combined with UPRTase activity. Phase I clinical trials are ongoing.
  • the recommended dose of 5-FC for human use (200 mg/kg/day) needs to be administered to patients for 14 days.
  • Ancotil ® is currently available as 500 mg tablets or 1% intravenous solution bottles of 250 mL for systemic administration (or 250 mg and 500 mg capsules in the US under Ancobon ® trademark).
  • patients average weight 60 kg will have to take daily 12 g of 5-FC (4 intakes of 3.0 g each). This means that patients will need daily to swallow 24 Ancotil ® tablets (500 mg each) or to be injected with 1.2 L of the infusion solution, making the treatment not compliant with their welfare. Accordingly, there is a strong need to develop a new pharmaceutical formulation of 5-FC, much more patient friendly.
  • a new 5FC pharmaceutical formulation has been developed.
  • Main characteristics of pharmaceutical formulations are: (i) physical, such as size, hardness, friability, disintegration and dissolution; (ii) chemical, such as drug content; and (iii) stability and sensory, such as appearance, odor and taste. All of these three characteristics are equally important with respect to patient acceptance, preference and compliance. For example, drinkable formulation reduces patient acceptance, preference and compliance, especially for patients treated for cancer.
  • aspects of the present invention include pharmaceutical formulations that comprise at least one prodrug for oral administration useful in treating/preventing hyperproliferative diseases such as cancer, fungal and other disease.
  • the formulation of the Invention is particularly useful for improving patient compliance with administration by reducing the drug intake difficulty.
  • the prodrug is 5-fluorocytosine (5-FC).
  • compositions that comprise, consist essentially of, or that consist of granules comprising 5-fluorocytosine (5- FC).
  • these granules comprise a core matrix and a coating.
  • the core matrix comprises 5-fluorocytosine (5-FC) and at least one pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier included in the core matrix is a granulating agent.
  • the carrier amount in the core matrix can range from about 5% to about 40%, advantageously from about 10% to about 15%, by weight of the core matrix.
  • said carrier comprises microcrystalline cellulose and polyvinyl pyrrolidone.
  • said carrier comprises microcrystalline cellulose, polyvinyl pyrrolidone and polyvinyl acetate.
  • said carrier comprises microcrystalline cellulose and copovidone.
  • the weight ratio microcrystalline cellulose / copovidone is from about 2 to about 6, advantageously about 4.
  • the core matrix of the granule formulation of the Invention comprises 5-fluorocytosine (5-FC), microcrystalline cellulose and polyvinyl pyrrolidone, and preferably further contains polyvinyl acetate.
  • the core matrix of the granule formulation of the Invention comprises 5-fluorocytosine (5-FC), microcrystalline cellulose and polyvinyl acetate, and preferably further contains polyvinyl pyrrolidone.
  • Certain aspects of the pharmaceutical formulations of the Invention concern a core matrix that further comprises a sustained release agent.
  • the sustained release agent is in an amount above 10 %, advantageously above 20%, by weight of the core matrix.
  • said sustained release agent is a mixture of polyvinyl acetate and polyvinyl pyrrolidone.
  • said sustained release agent is at least one component selected in the group consisting of high molecular weight hydroxypropylmethylcellulose (HPMC 1000 to 100 000), high molecular weight hydroxypropyl cellulose, ethylcellulose, high molecular weight polyethylene glycol, compritolTM, precinol, starch, carboxymethylcellulose, polyvinyl alcohol, methyl cellulose, alginate sodium or carraghenans.
  • the weight ratio polyvinyl acetate / polyvinyl pyrrolidone is 80%/20%.
  • the sustained release agent is Kollidon SR (BASF-Laserson).
  • compositions that comprise one first coating.
  • the first coating comprises hydrophilic polymer.
  • the first coating comprises hydroxypropylmethylcellulose
  • the first coating further comprises at least one lubricant.
  • the first coating further comprises stearic acid.
  • the first coating comprises, consists essentially of, or consists of hydroxypropylmethylcellulose (HPMC) and stearic acid.
  • HPMC hydroxypropylmethylcellulose
  • the first coating composition comprises at least 10%, advantageously at least 14%, or at least 30%, by weight of stearic acid.
  • the first coating composition comprises 30% of stearic acid.
  • the first coating composition is Sepifilm LP030.
  • compositions that further comprise one second coating.
  • the second coating composition comprises at least one water soluble polymer.
  • the said water-soluble polymer is selected in the group consisting in polyvinylpyrrolidone, polyvinylpyrrolidone vinyl acetate, hydroxypropylmethyl cellulose and polyethylene glycol.
  • the said water-soluble polymer comprises polyvinylpyrrolidone and polyethylene glycol.
  • the po ly vi ny lpyrrol idonc is PVP K 1 7 or K90.
  • the polyethylene glycol is PEG4000.
  • the water-soluble polymer comprises polyvinylpyrrolidone and polyethylene glycol in weight ratio 10/3 (w/w), respectively.
  • the granule formulation comprises at least 1000 mg, preferably at least 1500 mg, of 5-FC.
  • the granules show a 5-FC dissolution rate defined as follows:
  • 15 min at least 80%, in particular 80%-95% (preferably about 81.00 %).
  • the granules exhibit a 5-fluorocytosine (5-FC) dissolution rate comparable to the commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets (500 mg).
  • Preferred methods of making 5FC pharmaceutical formulation involve over- granulation step.
  • Figure 1 Comparative analysis of batch 100255 to reference Ancotil® dissolution profiles
  • Figure 5 Comparative analysis of batch 100286 to reference Ancotil®
  • Figure 6 Comparative analysis of batches 100283, 100315 and 100317 and reference Ancotil ® tablets
  • Figure 8 Comparative analysis of batch 100333 to reference Ancotil® pH 6.8 and 4.5
  • Figure 9 Redispersion analysis, batch 100341
  • Figure 12 Comparative analysis of batch 100356 to reference Ancotil® batch 80026952, pH 1.2 and 6.8 (mean values)
  • Figure 13 Comparative analysis of batch 100356 to reference Ancotil® batch 80026952, pH 4.5 and 6.8 (mean values)
  • Figure 14 Comparative analysis of batch 100397 to reference Ancotil® pH 4.5 and
  • Figure 15 Comparative analysis of batches 100356 and 100397, pH 4.5 and 6.8
  • Figure 16 Particle size distribution analysis (sieving) of batch 100412
  • Figure 17 Particle size distribution analysis (sieving) of batch 100413
  • Figure 19 Comparative analysis of batch 100412 to reference Ancotil®, pH 4.5 and
  • Figure 20 Comparative analysis of batch 100413 to reference Ancotil®, pH 4.5 and 6.8 (mean values)
  • Figure 22 Particle size distribution analysis (sieving) of batch 100426
  • Figure 23 Particle size distribution analysis (sieving) of batch 100428
  • Figure 24 Redispersion analysis, batches 100397, 100426 and 100428
  • Figure 27 Comparative analysis of batch 100426 to reference Ancotil®, pH 4.5 and 6.8 (mean values)
  • Figure 28 Particle size distribution analysis (sieving) of batch 100333
  • Figure 29 Particle size distribution analysis (sieving) of batch 100432
  • Figure 30 Redispersion analysis, batches 100432 and 100433
  • Figure 32 Comparative analysis of batch 100432 to reference Ancotil®, pH 4.5 and 6.8 (mean values)
  • Figure 34 Particle size distribution (sieving) of batch 100412
  • Figure 37 Comparative analysis of batch 100434 to reference Ancotil®, pH 4.5 and 6.8 (mean values)
  • the present invention concerns in a first aspect a granule formulation comprising 5- fluorocytosine wherein said granule comprises :
  • the core matrix comprises 5-fluorocytosine (5-FC) and at least one pharmaceutically acceptable carrier.
  • the coating comprises a moisture barrier.
  • the core matrix comprises 5-fluorocytosine (5- FC) and at least one pharmaceutically acceptable carrier, wherein said pharmaceutically acceptable carrier can be selected in the group consisting of solid diluents, disintegrants, binding agents, granulating agents, and optionally other formulating agents known in the art such as, for example, suspending agents, dispersing agents, solvents, preservatives or lubricants.
  • pharmaceutically acceptable carrier can be selected in the group consisting of solid diluents, disintegrants, binding agents, granulating agents, and optionally other formulating agents known in the art such as, for example, suspending agents, dispersing agents, solvents, preservatives or lubricants.
  • said pharmaceutically acceptable carriers are granulating agents selected from the group consisting in cellulose derivatives, microcrystalline cellulose, gelatin, starches, sugar (e.g. glucose, sucrose, lactose, trehalose, maltose), polyols (e.g. sorbitol, mannitol), polyvinylpyrrolidone (polyvidone), copolymers of vinylpyrrolidone and vinylacetate (also referred to as copovidone), gums (e.g. tragacanth, xantham, acaciae), sodium alginate or combinations thereof.
  • sugar e.g. glucose, sucrose, lactose, trehalose, maltose
  • polyols e.g. sorbitol, mannitol
  • polyvinylpyrrolidone polyvidone
  • copolymers of vinylpyrrolidone and vinylacetate also referred to as copovidone
  • gums e.g. trag
  • the core matrix of the granule formulation of the Invention comprises 5-fluorocytosine (5-FC), microcrystalline cellulose and a cop o lymer o f vinylpyrrolidone and vinylacetate (also referred to as copovidone, e.g. Plasdone® S-630, a synthetic, 60:40, linear, random copolymer of N-vinyl-2-pyrrolidone and vinyl acetate).
  • the carrier amount in the core matrix can range from about 5% to about 50%, especially from about 10% to about 20%, more particularly from about 10% to about 15%, by weight of the core matrix.
  • the weight ratio microcrystalline cellulose (e.g. Vivapur®) / copovidone is from about 2 to about 6, preferably about 4.
  • Preferably water is used in conjunction with the core matrix components as a wetting agent.
  • the granule formulation further comprises one first coating which comprises a water soluble coat former.
  • the first objective of this coating is to slow-down the 5-FC release from the granule for obtaining a dissolution rate profile comparable to the commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets.
  • said first coating composition comprises at least one binder chosen from starches, modified starches and the like, one or more compounds selected in the group consisting of natural wax either from vegetable or animal origin or synthetic wax, hydrophilic p o lymer such as hydroxypropylce llulo s e (HP C) , hydroxypropylmethylcellulose (HPMC - e.g. hypromellose 2910), hydroxypropylethylcellulose (HPEC), ethylcellulose, methylcellulose, cellulose acetate phthalate (CAP), microcrystalline cellulose or carrageenan, and mixtures thereof.
  • hydrophilic p o lymer such as hydroxypropylce llulo s e (HP C) , hydroxypropylmethylcellulose (HPMC - e.g. hypromellose 2910), hydroxypropylethylcellulose (HPEC), ethylcellulose, methylcellulose, cellulose acetate phthalate (CAP), microcrystalline cellulose or carrageen
  • said first coating composition further comprises at least one hydrophobic compound chosen from the group consisting of magnesium stearate, aluminium stearate, calcium stearate, stearic acid, talc, hydrogenated castor oil, and mixtures thereof.
  • at least one hydrophobic compound chosen from the group consisting of magnesium stearate, aluminium stearate, calcium stearate, stearic acid, talc, hydrogenated castor oil, and mixtures thereof.
  • one hydrophobic compound is sufficient, namely stearic acid benefits the manufacture of the particular granule formulation of the Invention.
  • said first coating composition comprises hydroxypropylmethylcellulose (HPMC) and stearic acid. It may further contain microcrystalline cellulose. According to preferred embodiment, said first coating composition comprises at least 10% of stearic acid, preferably at least 14% of stearic acid, and more preferably at least 30%> by weight of stearic acid. In particular embodiment, it comprises 30% by weight of stearic acid.
  • said first coating composition shows pH- independent solubility.
  • said first coating composition is a mixture of hydroxypropylmethylcellulose (also referred to as hypromellose), microcrystalline cellulose, stearic acid, and optionally other components such as pigments and lakes.
  • hydroxypropylmethylcellulose also referred to as hypromellose
  • microcrystalline cellulose stearic acid
  • pigments and lakes optionally other components such as pigments and lakes.
  • SepifilmTM LP030 SepifilmTM LP030 (Seppic).
  • the granule formulation of the Invention is meant to be dispersed in a glass of water prior to administration to the patient.
  • the 5-FC release from the granules during the reconstitution step i.e. aqueous dispersion before administration period of 5 to 15 minutes
  • the reconstitution step i.e. aqueous dispersion before administration period of 5 to 15 minutes
  • the core matrix may further comprise a sustained release agent, preferably a pH independent one.
  • this sustained release agent is in an amount above 10 % by weight of the core matrix, preferably above 20%, most preferably above 30 %, by weight of the core matrix.
  • said sustained release agent is a mixture of polyvinyl acetate and polyvinyl pyrrolidone.
  • said sustained release agent is at least one component selected in the group consisting of high molecular weight hydroxypropylmethylcellulose (HPMC 1000 to 100 000), high molecular weight hydroxypropyl cellulose, ethylcellulose, high molecular weight polyethylene glycol, compritolTM (a mixture of Glyceryl Dibehenate (CAS Number 94201-62-4 ), Tribehenin (CAS number 18641-57-1), and Glyceryl Behenate (CAS number 30233-64-8), precinol, starch, carboxymethylcellulose, polyvinyl alcohol, methyl cellulose, alginate sodium or carraghenans.
  • HPMC 1000 to 100 000 high molecular weight hydroxypropylmethylcellulose
  • ethylcellulose high molecular weight polyethylene glycol
  • compritolTM a mixture of Glyceryl Dibehenate (CAS Number 94201-62-4 ), Tri
  • the weight ratio polyvinyl acetate / polyvinyl pyrrolidone is 80%/20%.
  • said sustained release agent in the core matrix is a spray dried, non-hygroscopic powder consisting of polyvinyl acetate (8 parts w/w) and polyvinyl pyrrolidone (2 parts w/w).
  • a specific example is Kollidon® SR.
  • the preferred granule formulation of the Invention comprises approximately in the core matrix :
  • microcrystalline cellulose - 10.8% by weight of microcrystalline cellulose
  • non-hygroscopic powder consisting of polyvinyl acetate (8 parts w/w) and polyvinyl pyrrolidone (2 parts w/w)(e.g. kollidon® SR), and
  • the granule formulation of the invention further comprises a second coating.
  • This second coating objective is to increase the wettability of the granules of the Invention, and thus to enhance the granule water-redispersion characteristics, because the developed granule formulation is meant to be dispersed in a glass of water prior to administration to the patient.
  • 5-FC 5-fluorocytosine
  • Ancotil ® tablets it was requested for this granule formulation that 5-FC release from the granules during the reconstitution step (i.e. before administration - period of 5 to 15 minutes) should be avoided or as limited as possible.
  • said second coating composition comprises at least one water soluble polymer, i.e. polymeric composition, soluble in an aqueous solution.
  • said water-soluble polymer is selected in the group consisting in modified starch, gelatin, polyvinylpyrrolidone, copolymers of vinylpyrrolidone and vinylacetate (copovidone), cellulose derivatives (such as for example hydroxypropyl methylcellulose (HPMC) and hydroxypropyl cellulose (HPC)), polyethylene glycol, polyvinyl alcohol and mixtures thereof.
  • said water-soluble polymer is selected in the group consisting in polyvinylpyrrolidone and polyethylene glycol.
  • said water-s o lub l e p o lymer i s polyvinylpyrrolidone.
  • said water-soluble polymer is polyethylene glycol.
  • said water-soluble polymer comprises polyvinylpyrrolidone and polyethylene glycol.
  • the term "polyvinylpyrrolidone” or PVP refers to any of the polymers of vinylpyrrolidone, or derivatives thereof. While PVP is typically made by a free radical polymerization process, any soluble grade polymer of vinylpyrrolidone can be used according to the invention. Typically, linear PVP polymers are water soluble and cross-linked PVP polymers are not water soluble.
  • PVP is selected in the group consisting of grade K12, K15, K17, K25, K30, K60, and K90. According to preferred embodiment, it is PVP K17 or K90.
  • PVP Kn refers to a PVP with a K value of n.
  • K-value of a polymer is a well- known constant, which is function of the average degree of polymerization and the intrinsic viscosity of the polymer, and may be determined easily by one skilled in the art by viscosity measurements.
  • polyethylene glycol refers to any of the polymer of ethylene oxide or derivatives thereof.
  • PEG is the range of the 300-20000, more particularly it is selected in the group consisting of PEGIOOO, PEG1500, PEG2000, PEG4000, PEG6000 and PEG20000. According to preferred embodiment, it is PEG4000.
  • PEGn refers to a PEG having an average molecular weight of n.
  • said water-soluble polymer comprises polyvinylpyrrolidone and polyethylene glycol in ratio 10/3 w/w, respectively.
  • the granule formulation comprises at least 1000 mg, preferably at least 1500 mg, of 5-FC.
  • the preferred granule formulation of the Invention comprises approximately:
  • the granule formulation of the invention shows a 5-FC pH-independent solubility.
  • the granule formulation according to the invention shows a 5-FC dissolution rate defined as follows:
  • the dissolution is carried out by using USP type II apparatus with a paddle rotating speed of 75 rpm in 500 ml NaCl pH 6.8 and the time percent 5-FC release of the granule formulation of the Invention is :
  • the dissolution is carried out by using eq. 3g of 5-FC, USP type II apparatus with a paddle rotating speed of 75 rpm, 37°C, in 500 ml NaCl pH 4.5 and the time percent 5-FC release of the granule formulation of the Invention is :
  • 15 min at least 80%, in particular 80%-95% (preferably about 81.00 %).
  • the granule formulation according to the invention shows a 5-FC dissolution rate defined as follows:
  • the dissolution is carried out by using USP type II apparatus with a paddle rotating speed of 75 rpm in 500 ml NaCl pH 6.8 and the time percent 5-FC release of the granule formulation of the Invention is : 2 min: 15%-25% (preferably about 17.50 %),
  • the dissolution is carried out by using eq. 3g of 5-FC, USP type II apparatus with a paddle rotating speed of 75 rpm, 37°C, in 500 ml NaCl pH 4.5 and the time percent 5-FC release of the granule formulation of the Invention is :
  • the granule formulation according to the invention exhibits a 5-fluorocytosine (5-FC) dissolution rate comparable to that of a matricial tablet comprising 500 mg 5-FC as an active ingredient, the matrix preferably comprising at least one binding agent (such as PVP and/or microcrystalline cellulose), a diluent (such as corn starch) and at least one lubricant (such as magnesium stearate and/or hydrated precipitated colloidal silica). More preferably, the granule formulation according to the invention exhibits a 5-fluorocytosine (5- FC) dissolution rate comparable to that of a matricial tablet comprising or consisting of:
  • This composition is the composition of the commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablet.
  • the granule formulation according to the invention thus preferably exhibits a 5-fluorocytosine (5-FC) dissolution rate comparable to that of the commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablet.
  • said 5-fluorocytosine (5-FC) granule dosage form has a bioavailability equivalent to that of a matricial tablet as defined above, in particular the commercially available reference 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets.
  • the Invention provides a granule formulation showing similar 5-FC dissolution rate than a matricial tablet as defined above, in particular reference Ancotil ® tablets (see Experimental section).
  • the granule formulation according to the invention may show similar 5- FC dissolution rate than a capsule comprising 250 to 500 mg of 5-FC as an active ingredient, mixed with corn starch, lactose and talc, the capsule shells preferably comprising parabens (butyl, methyl, propyl) and sodium propionate and a coloring agent.
  • This is the composition of commercially available reference Ancobon ® capsules.
  • the granule formulation according to the invention thus preferably exhibits a 5-fluorocytosine (5-FC) dissolution rate comparable to that of the commercially available 5-fluorocytosine (5-FC) capsules i.e. Ancobon ® capsules (250 or 500 mg).
  • “comparable” or “similar” dissolution rate is meant a dissolution rate from the granule formulation into liquid phase, measured under the same conditions, which is at least 80% of the dissolution rate of commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets (500mg), preferably at least 90% of the dissolution rate of commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets (500mg), even more preferably at least 100%) of the dissolution rate of commercially available 5-fluorocytosine (5-FC) tablet i.e. Ancotil ® tablets (500mg).
  • Ancotil ® tablets and the Ancobon ® capsules being commercially available, it is within the reach of the one skilled in the art to compare dissolution rate of this reference compound with any other tested compound in comparable conditions.
  • the invention also relates to a granule formulation comprising 5- fluorocytosine wherein said granule comprises:
  • Such a granule formulation may further have any feature or combination of features as defined above.
  • the present invention further concerns composition comprising the said granule formulation.
  • the invention also provides methods for preparation of the granules and compositions comprising the granules.
  • the granules of the Invention in particular the preferred ones, can conveniently be manufactured by a wet granulation route, followed by dry blending and coating steps.
  • "Granulation” is commonly defined as a size-enlargement process in which small particles are gathered into larger, permanent aggregates in which the original particles can still be identified.
  • Weight granulation is a variation on this process, as refers to a granulation that adds solvents and binders to the enlargement process (see for example Lipps (1993) J. Pharm. Sci. 83: 937-947).
  • a variety of blending, or mixing, or granulating, apparatus are commonly available.
  • the granulation can be done on a Fluid Bed Granulator, (e.g. Glatt Air Techniques Inc., N. J.).
  • the granulation is done on high shear granulator (e.g. GLATT TMG).
  • the said granulator activity can further be completed by using granule drying apparatus, such as a fluidized bed drier (e.g. GLATT GPCG1) and a granule coating apparatus, (e.g. GLATT GPCG1).
  • a fluidized bed drier e.g. GLATT GPCG1
  • a granule coating apparatus e.g. GLATT GPCG1
  • the temperature during granulation can be set at any point as long as it does not exceed the melting point of any components in the formulation and the balance between spraying and drying is kept. Typically, the temperature is set in the range of 20° C to 50°C.
  • the granule formulation of the invention are prepared by high shear wet granulation (HS WG) process, and more preferably said process comprises at least one step of over-granulation.
  • HS WG high shear wet granulation
  • Over granulation of the formulation of the Invention is obtained by adding an excess of granulating solution after the point at which granulation of the blend is achieved. Over granulation allows obtaining granules with a greater particle size.
  • the granule formulation of the invention present a particle size distribution such that at least 40% (in weight) of the particles present a particle size above 500 ⁇ , preferably above 700 ⁇ ; in some embodiments said particle size distribution is such that at least 50% (in weight) of the particles present a particle size above 500 ⁇ , preferably above 700 ⁇ ; in some embodiments said particle size distribution is such that at least 60%> (in weight) of the particles present a particle size above 500 ⁇ , preferably above 700 ⁇ ; in some embodiments said particle size distribution is such that at least 70% (in weight) of the particles present a particle size above 500 ⁇ , preferably above 700 ⁇ ; in preferred embodiments said particle size distribution is such that at least 70% (in weight) of the particles present a particle size above 700 ⁇ .
  • the final pharmaceutical form of the formulation of the Invention is intended to be a sachet with granules to be reconstituted in liquid, preferably water, before administration, and flavouring can be added in order to taste-mask the bitterness of 5-fluorocytosine contained in the granules and to reach good patient compliance.
  • flavouring can be added in order to taste-mask the bitterness of 5-fluorocytosine contained in the granules and to reach good patient compliance.
  • a non- limiting representative list of examples includes toffee, orange, lime, lemon, grapefruit, pinenapple, or combination thereof.
  • said flavoring compound can represent about 1% to 2% of the final granule formulation.
  • the invention concerns the use of a granule formulation comprising
  • 5-fluorocytosine according to the invention for the preparation of a medicament for use in the treatment of fungal diseases, proliferative diseases, especially cancer or diseases associated to an increased osteoclast activity (e.g. rheumatoid arthritis, osteoporosis), infectious disease and other disease therapies, preferably for use in the treatment of cancer.
  • the present invention also relates to a granule formulation comprising 5- fluorocytosine (5-FC) according to the invention, for use in the treatment of fungal diseases, proliferative diseases, especially cancer or diseases associated to an increased osteoclast activity (e.g. rheumatoid arthritis, osteoporosis), infectious disease and other disease therapies, preferably for use in the treatment of cancer.
  • 5- fluorocytosine 5-FC
  • the present invention also concerns a kit of parts, comprising:
  • a polypeptide comprising cytosine deaminase activity a polypeptide comprising cytosine deaminase activity, a polynucleotide encoding a polypeptide comprising cytosine deaminase activity, or an expression vector to induce expression of a polypeptide comprising cytosine deaminase activity,
  • kits of part are also intended for use in the treatment of fungal diseases, proliferative diseases, especially cancer or diseases associated to an increased osteoclast activity (e.g. rheumatoid arthritis, osteoporosis), infectious disease and other disease therapies, preferably for use in the treatment of cancer.
  • the present invention further provides methods of treating a fungal infection by administering a granule formulation of the invention.
  • the present invention also provides a method of treating proliferative diseases, cancer, diseases associated to an increased osteoclast activity, infectious disease or other disease in a mammal by administering a granule formulation of the invention in conjunction with a polypeptide comprising cytosine deaminase activity or in combination with a polynucleotide encoding such a polypeptide.
  • the present invention concerns a method of treating a fungal disease.
  • the method comprises administering to a subj ect in need thereof a therapeutically effective amount of a granule formulation of the invention comprising 5- fluorocytosine.
  • the method can be used to treat any applicable fungal disease, including a fungal disease which is a brain fungal disease, an internal organ fungal disease, vaginal fungal disease, mouth fungal disease foot fungal disease and/or which is an infection by a fungus.
  • a fungal disease which is a brain fungal disease, an internal organ fungal disease, vaginal fungal disease, mouth fungal disease foot fungal disease and/or which is an infection by a fungus.
  • it can be beneficial to coadminister another anti-fungal agent.
  • Any appropriate antifungal agent may be coadministered with the granule formulation of the invention comprising 5- fluorocytosine, including amphotericin B and azole antifungals such as fluconazole and itraconazole.
  • additional agents may be formulated with the granule of the invention or administered separately.
  • the present invention concerns a method for treating a hyperproliferative disease.
  • the method comprises administering to a subject in need thereof a sufficient amount of (i) a polypeptide comprising cytosine deaminase activity or (ii) an expression vector to induce expression of a polypeptide comprising cytosine deaminase activity in hyperproliferating cells, and a therapeutically effective amount of a granule formulation of the invention comprising 5-fluorocytosine.
  • the method can be used to treat any applicable hyperproliferative disease such as cancer, restenosis, diseases associated to an increased osteoclast activity, rheumatoid arthritis, Crohn's disease, chronic obstructive pulmonary disease, benign prostate hyperplasia, and others.
  • a hyperproliferative disease such as cancer, restenosis, diseases associated to an increased osteoclast activity, rheumatoid arthritis, Crohn's disease, chronic obstructive pulmonary disease, benign prostate hyperplasia, and others.
  • a hyperproliferative disease with a granule formulation of the invention comprising 5-fluorocytosine
  • it can beneficial to coadminister another antiproliferative disease agent.
  • Any appropriate anti-proliferative disease agent may be coadministered with the 5- fluorocytosine containing formulation of the invention, including, but not limited to methotrexate, cyclophosphamide, leucovorin, steroids and cancer drugs.
  • the present invention concerns a method for treating cancer.
  • the method comprises administering to a subject in need thereof a sufficient amount of (i) a polypeptide comprising cytosine deaminase activity or (ii) an expression vector to induce expression of a polypeptide comprising cytosine deaminase activity in cells of the cancer and a therapeutically effective amount of a granule formulation of the invention comprising 5-fluorocytosine.
  • the method can be used to treat any applicable cancer, including brain cancer (e.g.
  • glioblastoma multiforme lung cancer, colon-rectum cancer, breast cancer, prostate cancer, urinary tract cancer, uterine cancer, lymphoma, oral cancer, pancreatic cancer, leukemia, melanoma, stomach cancer, ovarian cancer, cancers of the adrenal gland, bladder cancer, bone cancer, bone marrow cancer, cervix cancer, ganglia cancer, kidney cancer, liver cancer, salivary glands cancer, skin cancer (e.g., melanoma), spleen cancer, testis cancer, thymus cancer, and thyroid cancer.
  • bladder cancer bone cancer, bone marrow cancer, cervix cancer, ganglia cancer, kidney cancer, liver cancer, salivary glands cancer, skin cancer (e.g., melanoma), spleen cancer, testis cancer, thymus cancer, and thyroid cancer.
  • Cancers to be treated include solid tumors including metastases to a brain cancer, lung cancer, colon-rectum cancer, breast cancer, prostate cancer, urinary tract cancer, uterine cancer lymphoma, oral cancer, pancreatic cancer, leukemia, melanoma, stomach cancer and ovarian cancer.
  • a cancer with a formulation comprising 5- fluorocytosine of the invention, it can be beneficial to coadminister another anticancer agent.
  • any appropriate anti-cancer agent may be coadministered with the 5- fluorocytosine -containing formulation of the invention, including, but not limited to leukovorin, busulfan, cisplatin, mitomycin C carboplatin; antimitotic agents, such as colchicine, vinblastine, paclitaxel, and docetaxel; topo I inhibitors, such as camptothecin and topotecan; topo II inhibitors, such as doxorubicin and etoposide; RNA/DNA antimetabolites, such as 5- azacytidine, 5-fluorouracil and methotrexate; DNA antimetabolites, such as 5-fluoro-2 ' deoxy- uridine, hydroxyurea and thioguanine; EGFR inhibitors, such as IressaTM (gefitinib) and TarcevaTM (erlotinib) ; proteosome inhibitors; antibodies, such as campath, HerceptinTM (trastuzumab), A
  • the present invention concerns a method for treating diseases associated to an increased osteoclast activity.
  • the method comprises administering to a subject in need thereof a sufficient amount of (i) a polypeptide comprising cytosine deaminase activity or (ii) an expression vector to induce expression of a polypeptide comprising cytosine deaminase activity in targeted cells, and a therapeutically effective amount of a granule formulation of the invention comprising 5- fluorocytosine.
  • the method can be used to treat any applicable diseases associated to an increased osteoclast activity, including osteoporosis, hypercalcemia of malignancy, rheumatoid arthritis, tumor metastases and Paget's disease, periodontal disease, fibrous dysplasia, bone resorption by osteoclasts exceeds, bone formation by osteoblasts leading to decreased bone mass, skeletal fragility and bone fracture.
  • any applicable diseases associated to an increased osteoclast activity including osteoporosis, hypercalcemia of malignancy, rheumatoid arthritis, tumor metastases and Paget's disease, periodontal disease, fibrous dysplasia, bone resorption by osteoclasts exceeds, bone formation by osteoblasts leading to decreased bone mass, skeletal fragility and bone fracture.
  • an extended release formulation comprising 5- fluorocytosine
  • any appropriate of these agent may be coadministered with the granule formulation of the invention comprising 5- fluorocytosine, including, but not limited to biphosphonate, selective oestrogen receptor modulators (SERMs), parathyroid hormone (PTH) (e.g. teriparatide (Forteo)), strontium ranelate, DenosumabTM or calcitonin, and others or a combination thereof.
  • SERMs selective oestrogen receptor modulators
  • PTH parathyroid hormone
  • Formulateo teriparatide
  • strontium ranelate DenosumabTM or calcitonin, and others or a combination thereof.
  • Such additional agents may be formulated with the granule of the invention or administered separately.
  • the present invention concerns a method for treating infectious diseases.
  • the method comprises administering to a subject in need thereof a sufficient amount of (i) a polypeptide comprising cytosine deaminase activity or (ii) an expression vector to induce expression of a polypeptide comprising cytosine deaminase activity in infected cells, and a therapeutically effective amount' of a granule formulation of the invention comprising 5- fluorocytosine.
  • the method can be used to treat any applicable non-fungal infectious disease, including viral, bacterial and mycoplasma based diseases. Examples of such diseases include HIV infection, HBV infection, HCV infection, HPV infection, herpes viral infection, Tuberculosis.
  • an infectious disease with an extended release formulation comprising 5- fluorocytosine
  • another anti-infectious disease agent may be coadministered with the granule formulation of the invention comprising 5-fluorocytosine, including, but not limited to antibiotics, and antivirals such as valcyclovir, neverapin, anti-HIV drug combos, ribovirin and others.
  • additional agents may be formulated with the granule of the invention or administered separately.
  • the term 'therapeutically effective amount' means the amount of a prodrug and/or other biological substance necessary to induce a desired pharmacological effect.
  • the amount can vary greatly according to the effectiveness of a particular active substance, the age, weight, and response of the individual host as well as the nature and severity of the host's symptoms. Other factors that can be taken into account include the expression rate of a cytosine deaminase polynucleotide from a vector and the activity of a cytosine deaminase for 5-FC. Accordingly, there is no upper or lower critical limitation upon the amount of the prodrug or active substance.
  • the therapeutically effective amount to be employed in the methods of the disclosure can readily be determined by those skilled in the art.
  • said polynucleotide or expression vector encoding a polypeptide comprising cytosine deaminase activity can comprise natural or synthetic nucleotides, be an oligonucleotide, be RNA or DNA, and be single or double stranded.
  • the polynucleotide is delivered using an expression vector, and more particularly, a viral or viral derived expression vector.
  • the viral or viral derived expression vector can be replicating or non-replicating, especially in human cells, can be delivered as a viral particle or as polynucleotides encoding the viral vector, and can be an adenoviral vector, a measles vector, a herpesvector, a retroviral vector, a poxvirus vector (including animal pox or vaccinia derived vectors), a parvovirus vector or any other viral vector known to one skilled in the art.
  • the viral vector is a replication competent vector capable of infecting mammalian cells, particularly dividing cells (i.e. oncogenic vectors), and more specifically is replication competent poxviral vector.
  • the viral vector is oncolytic viral vector.
  • the oncolytic virus is selected from the group consisting of a Newcastle Disease Virus (NDV), a Mumps Virus, a Measles Virus, a Vesicular Stomatitis Virus, a Para-influenza Virus, an Influenza Virus, an Adenovirus, a Herpes I Virus, a Vaccinia Virus, and a Reovirus.
  • the poxvirus according to the invention is a Vaccinia virus strains Copenhagen or WR (see for example WO2009/065547, WO 2009/065546 or W09531105).
  • the prodrug 5-FC is converted to a cytotoxic drug by the action of enzymes inherent in a microorganism or which have been otherwise recombinantly introduced into an organism.
  • the yeast, or bacterial, cytosine deaminase converts the innocuous antibiotic prodrug 5-FC into the cytotoxic chemotherapeutic agent 5-fluorouracil (5-FU).
  • 5-FU cytotoxic chemotherapeutic agent 5-fluorouracil
  • Humans (and mammals in general) are not known to have a naturally occurring gene encoding an enzyme with significant cytosine deaminase activity.
  • Yeast and bacterial cytosine deaminase have gained recognition in the treatment of cancers using gene delivery and viral vectors for the delivery of the enzyme followed by treatment with 5-FC, which is then converted by the enzyme to a cytotoxic drug.
  • the cytosine deaminase polypeptide which will catalyze the conversion of 5- fluorocytosine to 5-fluorouracil includes those from bacteria and fungi, such as cytosine deaminase from E.coli or Saccharomyces cerevisiae as well as improved recombinant cytosine deaminases (e.g. WO 99/54481 and WO05/07857) can be used. According to special embodiment the cytosine deaminase activity is combined to UPRTase activity.
  • the methods described according to the present invention can be used in any mammalian species, including human, monkey, cow, sheep, pig, goat, horse, mouse, rat, dog, cat, rabbit, guinea pig, hamster and horse. Humans are preferred.
  • an granule formulation comprising 5- fluorocytosine of the invention can be administered to the host by an appropriate route, either alone or in combination with another drug.
  • a 'therapeutically effective amount' of granule formulation comprising 5- fluorocytosine of the invention is administered.
  • a 'therapeutically effective amount' sufficient to achieve the desired therapeutic effect under the conditions of administration such as an amount sufficient to reduce or eliminate a fungal infection, reduce or eliminate a nonfungal infectious disease, or to slow the growth, reduce or eliminate cancer cells or cells of a hyperproliferative disease.
  • the 5-FC formulation of the invention is used to treat a subject for 7 days out of every month for months or years.
  • the dose can be from about 1500 to 6000 mg administered 1-4 doses per day.
  • the 5-FC formulation of the invention is administered from about 1500 to 3000 mg one or more times per day.
  • the dose of 5-FC is adjusted based upon the activity of cytosine deaminase activity within a subject, tissue or cell.
  • Example 1 5-FC dissolution from commercially available reference Ancotif 500 mg tablets and Ancobon ® 500 mg capsules
  • Dissolution medium NaCl pH 1.2
  • Dissolution Ancotil batch 80026952. buffer pH 1.2. 500 ml. 75 rpm
  • Dissolution medium buffer pH 6.8
  • Dissolution Ancotil batch 80026952. buffer pH 6.8. 500 ml. 75 rpm
  • Dissolution medium buffer pH 4.5
  • Dissolution Ancotil batch 80026952. buffer pH4.5. 500 ml. 75 rpm
  • Dissolution medium NaCl pH 1.2
  • Stearic acid is an hydrophobic agent present in the Sepifilm TM LP excipients that allows to protect humidity- sensitive active pharmaceutical ingredients against moisture and also to slightly delay drug dissolution from coated cores (granules, tablets).
  • Blending step Add 5-fluorocytosine (sieved on 1 mm screen) and microcrystalline cellulose (sieved on 1mm screen) and mix 4 minutes in a high shear granulator GLATT TMG (1 liter tank, impeller speed 200 rpm and chopper speed 300 rpm).
  • Granulation step (GLATT TMG 1 liter tank):
  • Calibration step Granules of batch 100255 were calibrated manually (forced sieving) on a 2 mm screen. Batch 100257 was calibrated (forced sieving) manually on a 1.6 mm screen before the drying operation.
  • Figure 2 compares batch 100257 to reference Ancotil ® tablets analysed in similar conditions. The comparisons made on Figure 2 (mean values) clearly show that batch 100257 was much closer to reference Ancotil ® tablets than batch 100255. The pH-dependence of the dissolution profiles for batch 100257 was more marked than for batch 100255 and presented moreover the same pattern than reference Ancotil ® tablets.
  • Example 3 Influence of the amount of Sepifilm TM LP030 on 5-FC dissolution from coated granules
  • a 20%) w/w content of Sepifilm TM LP 030 was evaluated (vs. 15%> w/w for batch 100257 - see above).
  • the 5-FC granules were also calibrated (forced sieving/before drying) on a larger screen size (i.e. 2 mm vs. 1.6 mm for batch 100257) to evaluate the influence of granule size on 5-FC dissolution.
  • Blending step Add 5-fluorocytosine (sieved on 1 mm screen) and microcrystalline cellulose (sieved on 1 mm screen) and mix 4 minutes in a high shear mixer GLATT TMG (1 litter tank, impeller speed 200 rpm and chopper speed 300 rpm)
  • Granulation step (GLATT TMG 1 liter tank):
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes).
  • 5-FC dissolution rate from batch 100283 was evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Tests were carried out in 900 ml of deionised water, in 900 ml of a pH 1.2 buffer and in 900 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 grams of 5-FC. Comparisons were made to reference Ancotil ® tablets using the same dissolution test protocol.
  • Figure 3 compares batch 100283 to reference Ancotil ® tablets analysed in similar conditions (mean values).
  • Figure 4 compares 5-FC dissolution profile from batch 100283 and 100257 (mean values). This Figure shows that 5-FC drug dissolution from the prepared granules was found to be slower for batch 100283 than for batch 100257 in deionised water and at pH 1.2. This phenomenon was attributed to the higher amount of Sepifilm TM LP 030 present in formulation batch 100283 (i.e. 20% w/w) than in formulation batch 100257 (i.e. 15% w/w).
  • Example 4 Influence of the presence of a viscosity increasing agent in the extra- granular phase on 5-FC dissolution from coated granules
  • a viscosity increasing agent such as xanthan gum
  • This excipient would be mixed with the granules following the Sepifilm TM LP 030 coating.
  • Xanthan gum is a common pharmaceutical excipient used to increase formulation viscosity following its dispersion in water. In this trial, xanthan gum was mixed with the coated granules of batch 100257 in a 2:98 w/w ratio.
  • Blending step Add coated granules of batch 100257 and xanthan gum and mix 1 minute in a Turbula blender (1 liter tank, 44 rpm).
  • 5-FC dissolution rate from batch 100286 was evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Tests were carried out in 900 ml of deionised water, in 900 ml of a pH 1.2 buffer and in 900 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 grams of 5-FC. Comparisons were made to reference Ancotil ® tablets using the same dissolution test protocol. Discussion
  • Figure 5 compares batch 100286 to reference Ancotil ® tablets analysed in similar conditions (mean values). The comparisons made on this figure clearly indicate that formulation of batch 100286 show dissolution profiles quite different than reference Ancotil ® tablets, especially in a pH 6.8 buffer and in deionised water. At pH 1.2, 5-FC dissolution from the prepared granules was however very close to drug dissolution observed for reference Ancotil ® tablets.
  • Redispersion characteristics of batches 100306 and 100307 were shown to be better than for batches 100283, 100302 and 100303 indicating that sodium lauryl sulphate alone can enhance granule dispersion characteristics. These characteristics were however shown to be less interesting than for batch 100305. This clearly indicates that the combination of sodium lauryl sulphate with mannitol should be the preferred choice to enhance granule dispersion.
  • - Xanthan gum a viscosity increasing agent
  • 5-FC dissolution rate from batches 100315 and 100317 were evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Tests were carried out in 900 ml of deionised water. All analyses were carried out at 37°C in deionised water and on an equivalent of 3 grams of 5-FC. Comparisons were made to reference Ancotil ® tablets using the same dissolution test protocol (see Figure 6).
  • Example 6 Enhancement of the granules redispersion characteristics with a second coating using hydrophilic polymers Qualitative and quantitative composition
  • the povidone/polyethylene glycol ratio used for this trial is 10/3 w/w.
  • Blending step - Add 5-fluorocytosine (sieved on 1 mm screen) and microcrystalline cellulose (sieved on 1 mm screen) and mix 4 minutes in a high shear mixer GLATT TMG (4 liter tank, impeller speed 200 rpm and chopper speed 300 rpm).
  • GLATT TMG 4 liter tank, impeller speed 200 rpm and chopper speed 300 rpm.
  • Granulation step (GLATT TMG 4 liter tank) (see note 1 in part 6.7.1):
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes).
  • 5-FC dissolution rate from batch 100333 was evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Tests were carried out in 900 ml of deionised water, in 900 ml of a pH 1.2 buffer, in 900 ml of a pH 6.8 buffer, in 500 ml of a pH 4.5 buffer and in 500 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 grams of 5-FC. Comparisons were made to reference Ancotil tablets using the same dissolution test protocol.
  • Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 grams of 5-FC) in 120 ml of water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5 minutes, 10 minutes and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each sample withdrawal. This protocol was designed to mimic what would be done in the clinical trials or by the patient taking the 5-FC granule formulation.
  • the followng table shows the results obtained for batch 100333 :
  • Figure 8 compares batch 100333 to reference Ancotil tablets analysed in similar conditions (500 ml volume dissolution medium, mean values). This experiment shows that 5-FC dissolution from granules of batch 100333 is still too rapid when compared to reference Ancotil ® tablets and especially in deionised water (not shown) and in the pH 6.8 buffer.
  • Example 7 Influence on 5-FC dissolution from coated granules of the presence of an additional amount of stearic acid in the granule core and of the povidone grade used for the second coating
  • Blending step - Add 5-fluorocytosine and stearic acid (sieved on 1 mm screen) and mix during 2 minutes in a high shear mixer GLATT TMG (1 liter tank, impeller speed 200 rpm
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen
  • PVPK17/PEG4000 coating PVP 90/PEG4000 coating (Batch 100340) (Batch 100341)
  • 5-FC dissolution rates from batch 100340 and batch 100341 were evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Tests were carried out in 900 ml of deionised water for both batches. For batch 100341, additional dissolution rates were performed in 900 ml of a pH 1.2 buffer, in 900 ml of a pH 6.8 buffer, in 500 ml of a pH 4.5 buffer and in 500 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 grams of 5-FC. Comparisons were made to reference Ancotil ® tablets using the same dissolution test protocol.
  • Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 grams of 5-FC) in 120 ml of water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5 minutes, 10 minutes and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each withdrawal of sample. This protocol was designed to mimic what it would be done in the clinical trials or by the patient taking the 5-FC granule formulation.
  • Figure 10 compares (mean values) batch 100340 and batch 100341 to reference Ancotil ® tablets and batch 100333 analysed in similar conditions (900 ml volume dissolution media - deionised water).
  • Example 8 Influence of the presence and of the amount of Kollidon SR ® in the granule core on 5-FC dissolution from Sepifilm TM LP 030/povidone/polyethylene glycol coated granules
  • stearic acid as an hydrophobic agent inside the granules, at least at a 6% w/w ratio, having been shown to be unsuccessful with regards to slowing down 5-FC dissolution, it has been decided to evaluate the use of a specific pharmaceutical excipient aiming at sustaining the release of drugs.
  • the pharmaceutical excipient to be tested was Kollidon SR ® . This excipient is a co-processed mixture of 80% (w/w) of polyvinylacetate and 20% (w/w) of povidone K30. Polyvinylacetate is an insoluble polymer used in the development of extended release formulations. litative and quantitative composition
  • the povidone/polyethylene glycol ratio used for these trials is 10/3 w/w.
  • Blending step - Add 5-fluorocytosine, Kollidon SR ® and microcrystallme cellulose and mix 4 minutes in a high shear mixer GLATT TMG (1 liter tank, impeller speed 200 rpm and chopper speed 300 rpm).
  • Granulation step (GLATT TMG 4 liters tank) (see note 1 in part 6.9.1.1):
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes).
  • Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 grams of 5-FC) in 120 ml of water. The granules were stirred until a homogeneous 15 dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5, 10 and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each withdrawal of sample. This protocol was designed to mimic what would be done in a clinical trial or by the patient taking the 5-FC granule formulation.
  • FIGS 12 and 13 compares batch 100356 to reference Ancotil tablets (batch
  • batch 100356 demonstrated an interesting dissolution profile very close to reference Ancotil ® tablets in water and in a pH 6.8 buffer. In these medium, drug dissolution in the first 20 minutes was still however a little too fast when compared to the reference Ancotil ® tablets and attempts should be made to slightly slow down this dissolution rate. 5-FC dissolution from batch 100356 was found to be slower than reference Ancotil ® tablets in a pH 1.2 buffer. For dissolution trials carried out in 500 ml of dissolution media ( Figure 13), drug release from batch 100356 has been shown to be slower than the reference Ancotil ® tablets in the pH 6.8 buffer and faster in the pH 4.5 buffer. The dissolution profiles of batch 100356 have been shown in both test protocols to be greatly pH-dependent.
  • batch 100357 exhibited dissolution profiles that were very different compared to the reference Ancotil ® tablets (not shown). Under these analytical conditions, drug release from batch 100357 was faster than reference Ancotil ® tablets in the pH 6.8 buffer and more particularly in water where a significant difference could be observed. This difference in deionised water was actually greater for the batch 100357 than for the batch 100356, where 5- FC dissolution was found to be very close to the reference Ancotil ® tablets.
  • Batches 100356 and 100357 showed completely different behaviours by using the two dissolution protocols (i.e. either 900 ml or 500 ml of dissolution media), the first one being closer to the reference Ancotil ® tablets in tests carried out in 900 ml of dissolution media and the second being closer to reference Ancotil ® tablets in tests carried out in 500 ml of dissolution media.
  • the two dissolution protocols i.e. either 900 ml or 500 ml of dissolution media
  • Figure 14 compares batch 100397 to reference Ancotil ® tablets analysed in similar conditions (500 ml volume dissolution media, mean values).
  • batch 100397 presented dissolution profiles that were very close to reference Ancotil ® tablets.
  • 5-FC dissolution from batch 100397 in the pH 4.5 was slightly slower than the one observed with the reference Ancotil ® tablets but dissolution in the pH 6.8 was almost similar.
  • batch 100397 was actually the batch showing the closest dissolution profile to the reference Ancotil ® tablets.
  • Example 9 Optimization of the granulation process: introduction of Kollidon SR ® in the granulating solution
  • Blending step - Add 5-fluorocytosine, 1 ⁇ 2 Kollidon SR ® (sieved on 1mm screen) and microcrystalline cellulose and mix 4 minutes in a high shear mixer GLATT TMG (1 liter tank, impeller speed 200 rpm and chopper speed 300 rpm)
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes for batch 100412 and 15 minutes for batch 100413).
  • Particle size distribution analysis Particle size distribution, determined by sieving, for batches 100412 and 100413 (after the coating step) are given in Figures 16 and 17.
  • Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 g of 5-FC) in 120 ml of water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5, 10 and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each withdrawal of samples. This protocol was designed to mimic what it would be done in a clinical trial or by the patient taking the 5-FC granule formulation.
  • the following Figure 19 compares batch 100412 to the reference Ancotil ® tablets (batch 80026952) analysed in similar conditions (500 ml volume dissolution medium, mean values).
  • the following Figure 20 compares batch 100413 to reference Ancotil tablets (batch 80026952) analysed in similar conditions (500 ml volume dissolution media, mean values).
  • the granulation protocol using a part of the Kollidon SR ® in the granulating solution led to an acceleration of 5-FC dissolution out of the granules when compared to the dissolution profiles obtained for batch 100397 (see previous example) in similar conditions. It should be noted that, in contrast to what was done for batch 100397, the batches 100412 and 100413 were not over-granulated.
  • composition % mg/sachet % t % mg/sachet
  • Blending step - Add 5-fluorocytosine (sieved on 1 mm screen), Kollidon SR ® (sieved on 1mm screen) and microcrystalline cellulose (sieved on 1 mm screen) and mix 4 minutes in a high shear mixer GLATT TMG (1 liter tank, impeller speed 200 rpm and chopper speed 300 rpm)
  • Granulation step (GLATT TMG 4 liter tank):
  • Calibration step Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes).
  • Particle size distribution analysis Particle size distribution, determined by sieving, for batches 100397, 100426 and 100428 (after the coating step) are given in Figures 21, 22 and 23.
  • Dissolution rate analysis 5-FC dissolution rate from batches 100397, 100426 and 100428 was evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm. Trials were carried out in 500 ml of a pH 4.5 buffer and in 500 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 grams of 5-FC. Comparisons were made to the reference Ancotil ® tablets using the same dissolution test protocol.
  • Redispersion trials were carried out by dispersing the formulations (for an equivalent of 1.5 g of 5-FC) in 120 ml of tap water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5, 10 and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each withdrawal of sample. This protocol was designed to mimic what it would be done in a clinical trial or by the patient taking the 5-FC granule formulation.
  • the next trial consisted in repeating the manufacturing of batch 100397 without over- granulating the powder during the granulation step. This trial was carried out in order to evaluate if the over-granulation process was necessary to achieve the desired dissolution profile or if this was only formulation- dependent.
  • Figure 27 compares batch 100426 to reference Ancotil ® tablets analysed in similar conditions (500 ml volume dissolution media, mean values).
  • Formulation batch 100426 has been shown to present, the closest dissolution profiles to the reference Ancotil ® tablets.
  • This step was carried out to evaluate the over-granulation process for a batch composition that does not contain Kollidon SR ® as a sustained release agent in the granule core.
  • Blending step Add 5-fluorocytosine (sieved on 1 mm screen) and microcrystalline cellulose (sieved on 1mm screen) and mix 4 minutes in a high shear mixer GLATT TMG (4 liters tank, impeller speed 200 rpm and chopper speed 300 rpm).
  • GLATT TMG 4 liters tank, impeller speed 200 rpm and chopper speed 300 rpm.
  • Granules were calibrated manually (forced sieving) on a 2 mm screen.
  • Particle size distribution analysis Particle size distribution, determined by sieving, for batches 100333 and 100432 (after the coating step) are given in Figures 28 and 29.
  • Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 grams of 5-FC) in 120 ml of tap water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained. Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5, 10 and 15 minutes. The suspension was allowed to settle between each time points and was re-homogenized (spatula) before each withdrawal of the samples. This protocol was designed to mimic what it would be done in a clinical trial or by the patient taking the 5-FC granule formulation.
  • Figure 31 compares batch 100432 to batch 100333 analysed in similar conditions (500 ml volume dissolution media, mean values). Both batches have the same qualitative and quantitative compositions but, on the contrary to the batch 100333, the batch 100432 was over- granulated.
  • composition % mg/sachet % mg/sachet
  • Blending step Add 5-fluorocytosine (sieved on 1 mm screen), 1 ⁇ 2 Kollidon SR ® (sieved on 1 mm screen) and microcrystalline cellulose (sieved on 1 mm screen) and mix 4 minutes in a high shear mixer GLATT TMG (1 liter tank, impeller speed 200 rpm and chopper speed 300 rpm).
  • Granules were calibrated manually (forced sieving) on a 2 mm screen (time of operation: 10 minutes for batch 100412 and 15 minutes for batch 100434).
  • Particle size distribution analysis Particle size distribution, determined by sieving, for batch 100412 and batch 100434 (after the coating step) are given in Figures 33 and 34.
  • Dissolution rate analysis 5-FC dissolution rate from batches 100434 and 100412 was evaluated using a USP type II apparatus with a paddle rotating speed of 75 rpm in 500 ml of a pH 4.5 buffer and in 500 ml of a pH 6.8 buffer. All analyses were carried out at 37°C and on an equivalent of 3 g of 5-FC. Comparisons were made to the reference Ancotil ® tablets using the same dissolution test protocol. Redispersion analysis : Redispersion trials were carried out by dispersing the formulation (for an equivalent of 1.5 g of 5-FC) in 120 ml of water. The granules were stirred until a homogeneous dispersion using a spatula (equivalent to a spoon) was obtained.
  • Sample withdrawal was made at time 0 minute (i.e. just after dispersion) and after 5, 10 and 15 minutes.
  • the suspension was allowed to settle between each time points and was re-homogenized (spatula) before each sample withdrawal.
  • This protocol was designed to mimic what would be done in a clinical trial or by the patient taking the 5-FC granule formulation.
  • Figure 36 compares batches 100434 and 100412 analysed in similar conditions (500 ml volume dissolution media, mean values). Both batches have the same qualitative and quantitative compositions but, contrarily to the batch 100412, batch 100434 was over- granulated.
  • 5-FC dissolution from batch 100434 was similar to the dissolution profile of the reference Ancotil ® tablets at pH 6.8. At pH 4.5, 5-FC dissolution from batch 100434 was found to be slightly slower than for the reference Ancotil ® tablets.

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Abstract

L'invention concerne des formulations granulaires de 5-fluorocytosine (5-FC) et leur utilisation pour le traitement de maladies d'hyperprolifération, telles que le cancer ou une autre maladie, telle que des maladies fongiques. L'invention concerne en outre la fabrication desdits granules.
PCT/EP2012/062822 2011-07-01 2012-07-02 Formulations de 5-fluorocytosine et leurs utilisations Ceased WO2013004658A1 (fr)

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WO1992015308A1 (fr) * 1991-03-06 1992-09-17 The Wellcome Foundation Limited Utilisation de 5-fluoro-2'-desoxy-3'-thiacytidine pour le traitement de l'hepatite b
WO1995031105A1 (fr) 1994-05-13 1995-11-23 Thomas Jefferson University Procede permettant d'induire une reponse immunitaire au moyen de virus vaccinaux recombines
WO1999054481A1 (fr) 1998-04-17 1999-10-28 Transgene Mutant ayant une activite uracile phosphoribosyl transferase
WO2005007857A1 (fr) 2003-07-21 2005-01-27 Transgene S.A. Polypeptide a activite cytosine-desaminase amelioree
CN101288659A (zh) * 2007-04-18 2008-10-22 王雷波 一种胃内漂浮型微丸及其制备方法
WO2009065547A2 (fr) 2007-11-19 2009-05-28 Transgene Sa Vecteurs oncolytiques poxviraux
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WO2010002937A1 (fr) * 2008-06-30 2010-01-07 Tocagen Inc. Formulations de 5-fluorocytosine et leurs utilisations
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WO1992015308A1 (fr) * 1991-03-06 1992-09-17 The Wellcome Foundation Limited Utilisation de 5-fluoro-2'-desoxy-3'-thiacytidine pour le traitement de l'hepatite b
WO1995031105A1 (fr) 1994-05-13 1995-11-23 Thomas Jefferson University Procede permettant d'induire une reponse immunitaire au moyen de virus vaccinaux recombines
WO1999054481A1 (fr) 1998-04-17 1999-10-28 Transgene Mutant ayant une activite uracile phosphoribosyl transferase
WO2005007857A1 (fr) 2003-07-21 2005-01-27 Transgene S.A. Polypeptide a activite cytosine-desaminase amelioree
CN101288659A (zh) * 2007-04-18 2008-10-22 王雷波 一种胃内漂浮型微丸及其制备方法
WO2009065547A2 (fr) 2007-11-19 2009-05-28 Transgene Sa Vecteurs oncolytiques poxviraux
WO2009065546A1 (fr) 2007-11-19 2009-05-28 Transgene Sa Vecteurs oncolytiques dérivés de poxvirus
WO2010002937A1 (fr) * 2008-06-30 2010-01-07 Tocagen Inc. Formulations de 5-fluorocytosine et leurs utilisations
US20100029482A1 (en) * 2008-08-01 2010-02-04 Benko Zoltan L Use of 5-fluorocytosine as a fungicide

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