HK1227293A1 - Use of odiparcil in the treatment of mucopolysaccharidosis - Google Patents

Abstract

The present invention relates to a method of treatment of a mucopolysaccharidosis with 4-methyl-2-oxo-2H-1-benzopyran-7-yl-5-thio-ß-D-xylopyranoside.

Description

The present invention relates to the use of odiparcil, or a pharmaceutical composition containing this compound, in the treatment of mucopolysaccharidosis.

Technical background of the invention

Mucopolysaccharidosis (MPS) is a degenerative genetic disease associated with enzyme deficiency. In particular, MPS is caused by deficiency or inactivity of lysosomal enzymes that catalyze the progressive metabolism of complex sugar molecules called glycosaminoglycans (GAGs). These enzyme deficiencies result in an accumulation of GAG in the cells, tissues and especially the cell lysosomes of affected subjects, resulting in permanent and progressive cell damage that affects the appearance, physical abilities, organ functioning, and, in most cases, mental development of affected subjects.

Eleven distinct enzyme deficiencies have been identified, corresponding to seven distinct clinical categories of MPS. Each MPS is characterized by a deficiency or inactivity of one or more enzymes that degrade mucopolysaccharides such as heparanesulfate, dermatanesulfate, chondroitinesulfate, keratanesulfate.

Type III mucopolysaccharidosis (MPS III) or Sanfilippo disease is a lysosomal overload disease, of the mucopolysaccharidosis group, characterized by severe and rapid intellectual degradation. The first symptoms appear between 2 and 6 years of age: behavioural disturbances (hyperkinesia, aggressiveness) and intellectual degradation, sleep disturbances with very moderate dysmorphic signs. The neurological impairment becomes more marked around the age of 10 with loss of psychomotor acquisitions and communication with the environment. Epilepsy often occurs after the age of 10. The disease is due to the presence of hepase sulfate due to a deficiency of this type of transaminase or another enzyme responsible for the catabolism: alpha-Nicotensin type 4 (Nicotensin type II), sulfuric acid (Nicotensin type III), alpha-Nicotensin type 4 (Nicotensin type II), and sulfuric acid (Nicotensin type II), sulfuric acid (Nicotensin type II), and sulfuric acid (Nicotensin type II), which are not effective in the treatment of this disease.

Mucopolysaccharidosis type VI (MPS VI) or Maroteaux-Lamy disease is a lysosomal overload disease, of the group of mucopolysaccharidoses, characterised by severe somatic impairment and no psycho-intellectual regression. The prevalence of this rare mucopolysaccharidosis is between 1/250 000 and 1/600 000 births. In severe forms, the first clinical manifestations appear between 6 and 24 months and become more pronounced gradually: facial dysmorphia (macroglossia, constantly open mouth, thick features), joint limitations, very severe multiple dysplasia (platysomyosis, pectoris, scoliosis, valgus, carnatum, deformity of the lower bones), small size (inflammation of the heart valve), heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve, heart valve,The symptoms and severity of the disease vary greatly from one patient to another and there are also intermediate to very moderate forms (spondylo-epiphyseal-metaphyseal dysplasia associated with cardiorespiratory disorders). Like other mucopolysaccharidoses, Maroteaux-Lamy disease is related to a deficiency of the enzyme of mucopolysaccharide metabolism, in particular N-acetyl-galactosamine-fatase (also called the B-acetyl-metaphyseal enzyme).The enzyme deficiency blocks the progressive breakdown of dermatanesulfate, which leads to the accumulation of dermatanesulfate in the lysosomes of overloaded tissues. To date, there is only one drug approved for the treatment of this disease: Naglazyme® (human recombinant galsulfase) which is extremely expensive (in the United States, it is in the order of $350,000 per year).

Type VII mucopolysaccharidosis (MPS VII) or Sly's disease is a very rare lysosomal overload disease of the mucopolysaccharidosis group. The symptoms are extremely heterogeneous: antenatal forms (non-immune foetal-placental anasark), severe neonatal forms (with dysmorphia, hernias, hepatosplenomegaly, bogged feet, dysostosis, significant hypotonia and neurological disorders progressing to a natural stuttering and a profound intellectual disability in case of survival) and very moderate forms discovered in adolescence and even adulthood (thoracic cystitis). The disease is due to beta-D-glucanidase, responsible for the accumulation of various deficiencies in the lysosomes: chondroformis, chondroformis and chondroformis, not yet effective treatment of this disease, glyphosate and chondroformis.

Odiparcil (4-methyl-2-oxo-2H-1-benzopyran-7-yl-5-thio-β-D-xylopyranoside; CAS 137215-12-4) belongs to the thioxyloside family and is described in patent application EP-A-0 421 829. - What?

This compound was clinically developed (phases 1 and 2) in the treatment of thrombosis in the late 1990s and early 2000s. Its mechanism of action can be summarized as follows: Odiparcil acts as a substrate for an enzyme, GT1 (galactosyl transferase 1), which initiates the synthesis of GAG chains towards the dermatanesulfate/chondroitinesulfate pathway. These GAGs are constituents of the cell as proteoglycans (when bound to proteins on a primary serine and a sucrase that is xylose) and are also secreted into the extracellular membrane. They have a variety of coagulation control roles (paranasal/hepatal and secreted into the circulation of the dermatanesulfate) from the bloodstream to the bloodstream (xylose).

It has now been shown, and this is the subject of the present invention, that Odiparcil increases the synthesis of total GAG at the extracellular level and thereby will contribute to decreasing the intracellular load of GAG by acting as a lure, making the residual activity of N-acetylgalactosamine-4-sulfatase more effective.

Subject matter of the invention

The first aspect concerns Odiparcil for use in the treatment of mucopolysaccharidosis characterised by accumulation of chondroitin sulphate and/or dermatan sulphate.

Odiparcil and its production process are described in patent application EP-A-0 421 829.

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In one embodiment, the Odiparcil used in the present invention has at least 60%, preferably at least 70%, preferably at least 80%, preferably at least 90%, preferably at least 95%, preferably at least 98% or preferably at least 99% D configuration. In this embodiment, the Odiparcil is preferably in the form of a β-anomer.

In another embodiment, the Odiparcil used in the present invention is at least 60%, preferably at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or at least 99% as a β-anomer.

For example, approximately 100, 250, 300, 375, 400, 500, 750, 800, 1000, 1500, 2000, 3000, 4000 or 5000 mg of Odiparcil is given daily.

In one embodiment, at least about 0.1 mg to about 70 mg of Odiparcil per kg of patient body weight is administered daily, for example, at least about 1 or 2 mg, at about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 or 70 mg of Odiparcil per kg of patient body weight is administered daily.

In one embodiment, Odiparcil is administered once or twice daily (e.g. every 10 to 12 hours). Thus, the above mentioned daily doses can be divided for bi-daily (bid) administration, e.g. a daily dose of 1000 mg will be administered in two doses of 500 mg each. It is understood that each dose may consist of one or more pharmaceutical forms, e.g. a 500 mg dose may consist of two pharmaceutical forms of 250 mg each.

In one embodiment, Odiparcil is administered on an empty stomach (i. e. at least 1 hour before eating or more than 2 hours after eating) or with food (i. e. at the same time as or just before eating, e. g. about 20 to 30 minutes before or within 5 minutes after eating).

In one embodiment, Odiparcil is formulated in a pharmaceutical formulation containing one or more pharmaceutically acceptable excipients, using techniques well known to the trade, such as those described in Remington, The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, 2006 .

Thus, according to the second aspect, the invention relates to a pharmaceutical composition containing Odiparcil and one or more pharmaceutically acceptable excipients for use in the treatment of mucopolysaccharidosis characterised by accumulation of chondroitin sulphate and/or dermatan sulphate.

The pharmaceutical composition may be in any form adapted to the desired route of administration, which may be by bone, tongue, sublingual, oral, rectal, topical, intravenous, intra-arterial, subcutaneous, intranasal, transdermal, intramuscular or intraperitoneal.

In one embodiment, the pharmaceutical composition contains approximately 100 to 1000 mg of Odiparcil, e.g. 100, 125, 150, 250, 375, 400, 500 or 1000 mg of Odiparcil.

In one embodiment, the pharmaceutical composition is administered by injection, and comprises a vehicle which is typically a sterile aqueous solution sometimes containing, in addition to water, one or more ingredients such as sugars, preservatives, salts, tampons, etc. Injectable suspensions may include a suspension agent and a given liquid vehicle.

In one embodiment, the pharmaceutical composition is administered orally. Appropriate oral galenic forms include both solid and liquid formulations. When the pharmaceutical composition is a solid formulation (such as, for example, capsules, tablets, dry powders), useful excipients include, among others, diluents, lubricants, binders, disintegrating agents, fillers, etc. Solid formulations may be coated or uncoated; when they are, the coating may be enteric or non-enteric. When the pharmaceutical composition is a liquid formulation (such as, for example, a helix or syrup), useful excipients include, for example, water, glycols, a saline solution, aromatizing agents, etc.

In one embodiment, the composition is prepared by a wet granulation process, a technique well known to the trade. For example, Odiparcil, all or part of the diluent, the binder, and a sufficient amount of granulation fluid (such as water) are combined, granulated, dried, and ground to form granules. The granules are then combined or not with the rest of the liquids and the mixture is compressed. The compressed Odiparcil comprises about 5% to about 90% of the total compressed Odiparcil, compared to the compressed Odiparcil.

The invention is illustrated by the experimental part below.

Pharmacological activity 1. Results obtained on cells in culture 1.1. Endothelial cells from bovine aorta

Endothelial cells from bovine aorta (ECACC 92010601), cultured in 6-well plates, are incubated for 24 h in the presence of sodium 35S sulfate (10 μci/mL) and odiparcil soluble in DMSO at different concentrations (1-10 μM; 0.1% final DMSO). The culture surfactants are recovered and the cell carpets rinsed with phosphate buffer (PBS). The culture surfactants and rinsing solutions are grouped in tubes. An unlabeled deresulfate solution (200 μg) is then added to act as a trainer. The unincorporated 35S is then removed by filtration on Sephadex G25 columns.The samples are then centrifuged and the supernatant removed. The resulting precipitate is returned to solution in 2 M magnesium chloride and the GAGs are precipitated by 5 volumes of 95% ethanol. After centrifugation, the alcoholic precipitates are returned to solution in 0,9% sodium chloride and then the radioactivity is measured on a fraction after adding sparkling liquid in counting vials.

To type the GAGs produced in the culture cell supernatants, the alcohol precipitates returned to solution are treated with chondroitinase ABC (promeus vulgaris) at 0.5 mU/μL for 3 h at 37°C. After inactivation of the enzyme 3 min at 100°C, the undigested GAGs are precipitated by 5 volumes of 95% ethanol for 1 night at 4°C. After centrifugation the alcohol precipitates are returned to solution in 0.9% sodium chloride and then the radioactivity is measured on an aliquot fraction after addition of a scintillating liquid in counting vials.

For heparin sulphate-type GAGs, they are treated with heparinase II (Flavobacterium heparinum) at 4 mU/μL for 12 h at 30°C. After inactivation of the enzyme for 3 min at 100°C, the undigested GAGs are precipitated by 5 volumes of 95% ethanol for 1 night at 4°C. After centrifugation the alcoholic precipitates are reconstituted in 0.9% sodium chloride and then the radioactivity is measured on an aliquot fraction after addition of a sparkling liquid in vials.

As shown in Figure 1, Odiparcil increases dose-dependently the 35S-labelled GAG levels in bovine aortic endothelial cell culture surgeon.

In addition, enzymatic digestion indicates that the GAGs synthesised by cultured cells are predominantly of the chondroitin sulfate type.

1.2. Human fibroblasts

Normal human dermal fibroblasts (BIOAlternatives PF2) are cultured in 96-well plates for 24 h. The culture medium is then replaced with culture medium containing or not (control) Odiparcil at different concentrations (1 μM, 3 μM, 10 μM) or TGF-β reference at 10 ng/mL (positive control) and then the cells are incubated for 72 h with addition of the radioactive marker 3H-glucosamine for evaluation of total GAG synthesis. At the end of incubation, a chaotropic buffer is added to the culture plates to incorporate the fibroblasts. Total GAG from the cell units are then purified by liquid chromatography (Qi-colon lysis) and the radioactive activity is measured in the lysophore.

As shown in Figure 2, Odiparcil dose-dependently stimulates the synthesis of total GAG by human dermal fibroblasts (+94% at 10 μM). The data were analysed statistically by one-factor variance analysis followed by a Dunnett assay (* p< 0.05 vs control; ** p< 0.01 vs control; *** p< 0.001 vs control).

In vivo results obtained in rabbits after oral administration.

Odiparcil is administered orally at a dose of 400 mg/kg to the New Zealand rabbit. 4 h after administration the animals are anesthetized, and blood is drawn from citrated tubes after carotid artery catheterization. After centrifugation, plasma is collected and frozen. Plasma GAGs are isolated after protein digestion by Pronase E, for 48 h at 50°C. Proteins and protein residues are precipitated by addition of trichloracetic acid and incubation for 1 night at 4°C. After centrifugation the surfactants are recrypted, then dialysed 100 volumes of sodium chloride buffer, for 48 h at 4°C. A solution of 0.25 g of sodium chloride (0.1% diethyl chloride) is added to the final sample in order to precipitate the solution to 95% sodium chloride (GMP) and then the samples are precipitated to 2 volumes of sodium chloride (GMP) and dialysed at ambient temperature.

The plasma GAG extracts are quantified by measuring the uronic acid content, modified Bitter and Muir carbazole method. The qualitative analysis of plasma GAG extracts is performed by HPLC of disaccharides obtained after enzymatic digestion by Proteus vulgaris chrondroitinase ABC and Arthrobacter aurescens chrondroitinase AC.

The table below shows that treatment of animals with Odiparcil at a dose of 400 mg/ kg increases plasma GAG levels (as measured by uronic acid content) by a factor of 5 compared to control animals. Qualitatively, chondroitin-type GAGs have an increased galactosamine 6-sulfate component and dermatanesulfate component (chondroitin B) as measured by galactosamine 4-sulfate disaccharides (Δdi-4S DS). - What?

	µg UA/ml plasma	Δdi-0S (%)	Δdi-4S (%)	Δdi-6S (%)	Δdi-UA2S (%)	Δdi-4S DS (%)
Témoin	2,1	51,1	45,8	3,1	0	0
Odiparcil	11,4	18,6	26	30,8	4,1	20,5

The following is a list of the active substances in the active substance:

These results demonstrate that Odiparcil has the ability to increase the synthesis of total GAG (human fibroblasts), increase the concentration of extracellular chondroitin-type GAG (bovine aortic endothelial cells) and increase the synthesis of plasma GAG, particularly for chondroitin-type GAG.

Example of galenic formulation

Compact tablet obtained by a wet granulation process containing (% by weight): - What?

Odiparcil	90%
Cellulose microcrystalline (NF or Ph Eur)	7%
Povidone ou Polyvinylpyrrolidone (USP ou Ph Eur)	3%
Eau (USP ou Ph Eur)	qs pour granulation humide

Claims (8)

1. Odiparcil for use in the treatment of mucopolysaccharidosis characterised by accumulation of chondroitin sulphate and/ or dermatan sulphate.
2. Odiparcil for use as claimed 1, which is intended to be administered at approximately 100 mg to approximately 5000 mg daily.
3. Odiparcil for use as claimed 1 or claimed 2 which is intended for oral administration.
4. Odiparcil for use as claimed 3, which is intended to be taken with food.
5. A pharmaceutical formulation containing odiparcil and one or more pharmaceutically acceptable excipients, for use in the treatment of mucopolysaccharidosis characterised by accumulation of chondroitin sulphate and/or dermatan sulphate.
6. Pharmaceutical formulation for use as claimed 6, containing 100 mg to 1000 mg odiparcil.
7. Pharmaceutical formulation for use as claimed 5 or claimed 6, which is an oral galenic form, preferably a solid formulation.
8. Pharmaceutical formulation for use as claimed 7, which is a tablet.