HK1094650B - The use of fumaric acid derivatives in the manufacture of a medicament for treating asthma and chronic obstructive pulmonary diseases - Google Patents

Description

Use of fumaric acid derivatives for the production of medicaments for the treatment of asthma and chronic obstructive pulmonary disease

Technical Field

The invention relates to the use of fumaric acid derivatives for producing medicaments for treating cardiac insufficiency and asthma.

Background

Dialkyl and monoalkyl fumarates and salts thereof have been used successfully for the treatment of psoriasis for a prolonged period of time. This application has been described in numerous patents, see, for example, DE 2530372, DE 2621214 or EP-B-0312697.

In addition, the use of fumaric acid mono-and diesters for the treatment of autoimmune diseases, such as multiple arthritis or multiple sclerosis, has already been described (see DE 19721099.6 and DE 19853487.6) and in transplantation pharmaceuticals (see DE 19853487.6 and DE 19839566.3). Furthermore, the use of fumaric acid monoesters and diesters for the treatment of NF-. kappa.B-mediated diseases and for the treatment of mitochondrial diseases and/or as NF-. kappa.B inhibitors is known from DE 10101307.8 and DE 10000577.2. All publications mentioned describe fumaric acid monoesters and diesters, optionally in the form of certain salts.

Furthermore, the use of fumaric acid monoamides and diamides for the treatment of the indicated indications is known from DE 10133004.9. These amides are formed from amino acids, and preferably from specific peptides. Finally, fumaric acid oligomers and their use in the treatment of said diseases are known from DE 10217314.1.

Paroxysmal, significant respiratory distress is called asthma (bronchial asthma), which is suffered by about 4-5% of the population in industrial countries and has an increasing trend. This respiratory distress is due to a variable and reversible obstruction of the respiratory tract by an overreacting bronchial system, which is triggered by exogenous and/or endogenous stimuli. These stimuli include chemical or physical stimuli, infection, physical effort and/or emotional factors. After the disease has lasted for a long time, secondary diseases such as chronic bronchitis, emphysema, bronchiectasis, atelectasis or pulmonary heart disease or respiratory cardiac insufficiency (respiratory heart insufficiency) frequently occur.

Depending on the cause, variants of asthma are classified as asthma caused by allergy, infection, analgesics, working conditions or exertion, mixed forms of asthma or cardiac asthma (asthma cardiac), nasal asthma and uremic asthma (asthma uremicum). In particular, cardiac asthma can lead to respiratory distress due to increased obstruction of the pulmonary circulation in case of left ventricular insufficiency.

Now, in addition to proven methods of just avoiding the stimulation that triggers asthma, for example, β -2 sympathomimetics, corticosteroids, vago-mimetics, theophylline, anti-inflammatory agents, and anti-allergic agents are also administered in the medical treatment of asthma and/or in the alleviation of asthma.

At the molecular level, asthma appears to be characterized by an increased activity of Th2 lymphocytes in the lung, which in turn leads to an increased release of some Th2 cytokines, ultimately leading to the known features of asthma such as IgE isotype switching, mucus production and recruitment and activation of eosinophils. In addition, the Th2 cytokine appears to result in differentiation of other Th2 cells via a signal transduction pathway called JAK-STAT, resulting from an enhanced cycle. Increased proliferation of mesenchymal cells, particularly bronchial smooth muscle cells, is also observed.

The so-called JAK-STAT signal transduction pathway (Janus kinase signal transducer and activator of transcription pathway) is a pathway for transmitting information transmitted by signal peptides such as cytokines to the inside of cells and/or nuclei. Signal transduction occurs through STAT proteins, which are present in the cytoplasm and are initially inactive; there are 7 different STAT proteins known in humans. These STAT proteins are rapidly activated by phosphorylation, e.g., by janus kinases, due to binding of receptor ligands to the cell surface. Phosphorylation leads to homo-and heterodimerization of STAT proteins, which are rapidly transported into the nucleus where they bind to the target promoter and dramatically enhance the transcription rate of the promoter.

Acute or chronic inability of the heart to export blood for metabolism and/or to receive venous return under stress (stress insufficiency) or when it has rested (rest insufficiency) is called cardiac insufficiency. The insufficiency may be pure left or right ventricular insufficiency, but may also affect 2 ventricles.

The clinical signs of cardiac insufficiency can be attributed, as far as the etiology is concerned, to a number of causes, in particular inflammatory and degenerative changes of the myocardium and endocardium, coronary circulatory disorders, myocardial infarctions and injuries. Subsequently, cardiac insufficiency leads to peripheral circulatory pathologies, respiratory disorders, in particular cardiac asthma, renal insufficiency and electrolyte metabolism disorders and oedema and impaired skeletal muscle function (reduced functional capacity).

Indications are classified as acute cardiac insufficiency, energetic cardiac insufficiency (energetic-dynamic cardiac insufficiency) and debilitating cardiac insufficiency (also known as HEGGLIN syndrome II), excitatory motor cardiac insufficiency (excitatory motor cardiac insufficiency), cardiac insufficiency caused by arrhythmia (cardiac arrytomics), hypoxemia, latent, primary, compensatory, relative or stress insufficiency and/or left ventricular insufficiency.

At present, contractile substances are used for the pharmacological treatment of cardiac insufficiency, and glycosides (especially digoxin and digitoxin) are still used today for the treatment of chronic forms. However, in recent years vasodilators (nitro compounds and dihydralazine, alpha blockers, calcium antagonists and in particular ACE inhibitors) have become increasingly important. ACE inhibitors are most important for long-term therapy. In addition, diuretics are used. Catecholamines are used to treat acute forms, also amrinone may be used.

It is an object of the present invention to provide another medicament for the treatment of cardiac insufficiency and asthma. In particular, it is an object of the present invention to provide a therapeutic agent for cardiac asthma and left ventricular insufficiency in a field where they overlap each other. It is another object of the present invention to provide a therapeutic agent for two indications, either alone or in their overlapping fields, which is suitable for long-term treatment due to its good tolerability.

The object of the invention is achieved by the use of fumaric acid derivatives for the preparation of a medicament or pharmaceutical preparation for the treatment of asthma and/or cardiac insufficiency, in particular in humans.

Disclosure of Invention

According to a first aspect, the present invention relates to the use of a fumaric acid derivative selected from the group consisting of: dialkyl fumarates, monoalkyl fumarate salts, monoamides fumarates, monoamide fumarates, diamides fumarates, monoalkyl fumarates, carbocyclic or oxacarbocyclic oligomers of these compounds and mixtures thereof.

According to a second aspect, the invention relates to the use of fumaric acid derivatives selected from the group consisting of: dialkyl fumarates, monoalkyl fumarate salts, monoamides fumarates, monoamide fumarates, diamides fumarates, monoalkyl fumarates, carbocyclic or oxacarbocyclic oligomers of these compounds and mixtures thereof.

As described below and in the appended claims, the present invention is also directed to inhibiting bronchial smooth muscle cell uptake³Methods of H-thymidine, and methods of inhibiting the proliferation of these cells.

Finally, the invention relates to the use of the above fumaric acid derivatives for inhibiting PDGF-induced activation of STAT 1.

Drawings

The bar graph in FIG. 1 shows the degree of infarction after DMF administration, ischemia and control groups.

FIG. 2 shows PDGF-induction when DMF was added³Incorporation of H-thymidine into bronchial smooth muscle thinPercent inhibition in cells.

The bar graph in figure 3 shows the percentage of cell proliferation of bronchial smooth muscle cells stimulated with PDGF in the absence or presence of DMF and/or dexamethasone.

The bar graph in fig. 4 shows the end-diastolic diameter of the left ventricle in Dahl rats 8 weeks before and after the high-salt meal, in the absence or presence of DMF.

Detailed Description

According to one of its aspects, the present invention relates generally to the use of fumaric acid derivatives for the preparation of pharmaceutical formulations for the treatment of asthma and chronic obstructive pulmonary disease. Preferably, the asthma is caused by allergy, infection, pain relief, working conditions or exertion, particularly preferred is cardiac asthma.

According to another of its aspects, the invention also relates to the use of fumaric acid derivatives for the production of pharmaceutical preparations for the treatment or prophylaxis of cardiac insufficiency, myocardial infarction and angina pectoris. The cardiac insufficiency involved may be any type of cardiac insufficiency, regardless of its form and/or cause. Examples of cardiac insufficiency to be treated according to the invention are acute cardiac insufficiency, energetic cardiac insufficiency, energy-dynamic cardiac insufficiency and debilitating cardiac insufficiency (also known as HEGGLIN syndrome II), excitatory motor cardiac insufficiency, cardiac arrhythmia (cardiac arrhythmia) induced cardiac insufficiency, hypoxemia, potential, primary, compensatory, decompensatory, relative or stress insufficiency and/or left ventricular insufficiency, most preferably left ventricular insufficiency. The composition is also effective in preventing these diseases and/or myocardial infarction, including first, second or more infarctions.

These applications are based on the finding that fumaric acid derivatives inhibit PDGF- (platelet-derived growth factor) -induced activation of STAT 1. As described above, it is believed that STAT activation leads to a shift in cytokine patterns in asthma and ultimately to a shift in The vicious circle and mucus secretion, IgE production, and eosinophil recruitment with enhanced Th2 cell activity (a.b. pernis, p.b. rothman, "JAK-STAT signaling in asthma", The j.of clin. investigation, vol.10, No.1, May 2002).

The shift in the cytokine pattern described for fumaric acid derivatives in this document from Th1 to Th2 (see the aforementioned patent specification) would instead lead to the expectation of an enhancement of the vicious cycle. Therefore, they are not suitable for the treatment of asthma. Surprisingly, it was found that fumaric acid derivatives can inhibit the proliferation of respiratory smooth muscle cells. This appears to occur by inhibiting the PDGF-induced transcription factor STAT 1. It can be demonstrated in particular that fumaric acid derivatives can inhibit PDGF-induced STAT1 activation and PDGF-stimulated thymidine incorporation into BSM (bronchial smooth muscle) cells. Without being limited thereto, this proliferation inhibitory effect may be responsible for the effectiveness of the fumaric acid derivative in treating asthma.

The fumaric acid derivative to be used according to the present invention may be one or more selected from the group consisting of: dialkyl fumarates (i.e. dialkyl fumarates), monoalkyl hydrogen fumarates (i.e. monoalkyl fumarates), physiologically acceptable cations, especially alkali metal or alkaline earth metal cations or transition metal cations such as Li⁺、Na⁺、K⁺、NH₄ ⁺、Mg²⁺、Ca²⁺、Fe²⁺、Mn²⁺And Zn²⁺The salts of fumaric acid monoalkyl ester salts (i.e., fumaric acid monoalkyl ester salts), fumaric acid monoamides and fumaric acid diamides and their salts, carbocyclic and heterocyclic oligomers of these compounds, and mixtures thereof.

In a preferred embodiment, the fumaric acid derivative is selected from optionally substituted dialkyl fumarates and monoalkyl fumarates in the form of the free acid or salts thereof and mixtures thereof.

In this case, particular preference is given to the use of dialkyl fumarates of the formula (I) as described in DE 19853487.6,

wherein R is₁And R₂May be the same or different and independently represent C_1-24Alkyl or C_5-20Aryl, and these groups may optionally be substituted by halogen (F, Cl, Br, I), hydroxy, C_1-4Alkoxy, nitro or cyano. Particularly preferably, the dialkyl fumarate is dimethyl fumarate, diethyl fumarate and/or ethyl methyl fumarate.

Generally, according to the invention, alkyl is understood to mean a saturated or unsaturated, linear, branched or cyclic hydrocarbon radical having from 1 to 24 carbon atoms, which may optionally be substituted by one or more substituents. Preferably, alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, cyclopentyl, 2-ethylhexyl, hexyl, cyclohexyl, heptyl, cycloheptyl, octyl, vinyl, allyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2, 3-dihydroxypropyl, 2-methoxyethyl, methoxymethyl, 2-methoxypropyl, 3-methoxypropyl or 2, 3-dimethoxypropyl. Methyl or ethyl is most preferred.

According to the invention, aryl is to be understood as meaning optionally substituted aryl, alkyl-substituted aryl or aralkyl having from 5 to 20 carbon atoms, preferably aryl, alkyl-substituted aryl or aralkyl having from 6 to 10 carbon atoms. Exemplary groups are phenyl, benzyl, phenethyl, tolyl, ethylphenyl, propylphenyl, and butylbenzene, with tert-butylphenyl, phenyl, and benzyl being particularly preferred.

The substituents of said groups are preferably selected from the group consisting of: halogen (F, Cl, Br, I), hydroxy, C_1-4Alkoxy radical, C_1-4Alkyl, nitro and cyano.

Monoalkyl fumarates of the formula (II) as described in DE 19721099.6 can also be used advantageously,

wherein R is₁Is as defined above, A is hydrogen, an alkali metal or alkaline earth metal cation or a physiologically acceptable transition metal cation, preferably selected from Li⁺、Na⁺、K⁺、Mg²⁺、Ca²⁺、Zn²⁺、Fe²⁺And Mn²⁺N is equal to 1 or 2 and corresponds to the valence of A.

Exemplary compounds of formulae (I) and (II) are dimethyl fumarate, diethyl fumarate, ethyl methyl fumarate, monomethyl fumarate, monoethyl fumarate, calcium methyl fumarate, calcium ethyl fumarate, magnesium methyl fumarate, magnesium ethyl fumarate, zinc methyl fumarate, zinc ethyl fumarate, iron methyl fumarate and iron ethyl fumarate. They may be used alone or as a mixture.

Preferably, the fumaric acid amides to be used according to the invention are those described in DE 10133004.9. They correspond to the general formula (III),

wherein

R_aRepresents OR₃Or a D-or L-amino acid group-NH-CHR bound via an amide bond₄-COOH, wherein R₃Is hydrogen, straight or branched, optionally substituted C_1-24Alkyl, phenyl or C_6-10Aryl or aralkyl, and R₄Is the side chain of a natural or synthetic amino acid; and is

R_bIs represented byAmide-bonded D-or L-amino acid group-NH-CHR₅-COOH, wherein R₅Is the side chain of a natural or synthetic amino acid, or a peptide group having 2 to 100 amino acids, each of which may be the same or different, bonded through an amide bond.

The side chain of a natural or synthetic amino acid is typically a side chain selected from the group consisting of: ala, Val, Leu, Ile, Trp, Phe, Met, Tyr, Thr, Cys, Asn, Gln, Asp, Glu, Lys, Arg, His, citrulline, Hcy, Hse, Hyp, Hyl, Orn, Sar, and the side chain of Me-Gly. The side chains of Gly, Ala, Val, Ile, Leu and Me-Gly are preferred. If R is_aIs an L amino acid group-NH-CHR₄-COOH, and R_bIs an L-amino acid group-NH-CHR₅-COOH, then R₄And R₅May be the same or different. More preferably, R₄And R₅The same is true. Most preferably, R_aAnd R_bAre all glycine.

Or, R_aMay be a group OR₃And R is_bCan be L-amino acid group-NH-CHR₅-COOH or peptide group, R₅Have the meaning defined above. In this case, the fumaric acid derivative is a monoalkyl fumarate monoamide.

The peptide groups are bound by amide bonds and have from 2 to 100, preferably from 2 to 30, most preferably from 2 to 15 amino acids which may be the same or different. Peptide group R_bMost preferably selected from the group consisting of: peptide hormones, growth factors, cytokines, neurotransmitters, neuropeptides, antibody fragments, blood clotting factors and cyclosporine and derivatives and fragments thereof. Preferably, R_aIs methoxy or ethoxy, and R_bIs Gly, Ala, Val, Ile, Leu and Me-Gly.

The above-mentioned fumaric acid amides may be used singly or in admixture, or may be used in admixture with the above-mentioned monoalkyl or dialkyl fumarates.

Finally, as described in DE 10217314.1, it is also possible to use carbocyclic or oxacarbocyclic fumaric acid oligomers. They contain 2 to 10, preferably 2 to 6 and most preferably 2 to 3 units derived from fumaric acid and/or its esters and/or amides as described above as repeating units.

These fumaric acid oligomers are preferably obtained by (olefin) polymerization of the C-C double bond (for carbocyclic oligomers) and/or the C-C double bond and the carbonyl oxygen of the unit (for oxacarbocyclic oligomers). Preferably, the units derived from fumaric acid are derived from monomers selected from the group consisting of: fumaric acid and dialkyl fumarates as defined above, hydrogen monoalkyl fumarates, monoamides fumarates, diamides fumarates, monoalkyl monoamides fumarates and their salts and mixtures thereof. More preferably, the oligomer contains only units derived from 1 or 2 monomers. Most preferably, the oligomers contain only the same monomer units.

The carbocyclic oligomer consists of units derived from fumaric acid in such a way that these units are bonded to carbon atoms 2 and 3 of the fumaric acid backbone by covalent C-C bonds in such a way that a carbocyclic oligomer is formed. The oligomer backbone comprises an even number of carbon atoms and does not contain any other monomers and/or heteroatoms. The main chain is substituted at each carbon atom by one of the carboxylic acid and/or carboxylic acid amide groups of the fumaric acid monomer units constituting the main chain.

The oxacarbocyclic oligomer is composed of fumaric acid monomers in such a way that these units are bonded to one another at carbon atoms 1 and 3 via ether bridges. At the same time, atom C₂And C₃Is converted to C₁And C₂Is ethylenically unsaturated. Thus, in the case of oxacarbocyclic oligomers according to the invention, the ring contains polyoxypropylene units.

The term "oligomer" as used herein relates to a number of units of at least 2 fumaric acid monomers. Typically, the or carbocyclic fumaric acid oligomer contains 2 to 10, preferably 2 to 6 and most preferably 2 to 3 units derived from fumaric acid. Preferably, the carboxylic acid and/or carboxylic acid amide groups as substituents of the ring are all trans to each other.

In a preferred embodiment, a carbocyclic fumaric acid oligomer corresponding to the following formula (IVa) is used,

wherein the radical R_eAnd R_dIdentical or different and selected from amine groups (-NR)₁R₂) Amino acid group-NH-C (COOH) -R₅Peptide group having 2 to 100 amino acids, alkoxy group (-OR)₁) And hydroxy, R₁、R₂And R₅As defined above, and n is an integer from 2 to 10 inclusive, preferably from 2 to 6 inclusive.

Preferably, the group R_cAnd R_dEach independently is alkoxy or hydroxy, R_cAnd R_dIt is most preferred not to be hydroxyl at the same time. Thus, the monomer is preferably one or several monoalkyl hydrogen fumarates. In another embodiment, the group R_cAnd R_dMay each represent alkoxy-OR₁More preferably they are the same. In this case, the monomer is dialkyl fumarate.

Very preferably, r-1, t-2, c-3, t-4-tetrakis (methoxycarbonyl) cyclobutane or r-1, t-2, c-3, t-4, c-5, t-6-hexa (alkoxycarbonyl) cyclohexane is used according to the invention, preferably r-1, t-2, c-3, t-4-tetrakis (methoxycarbonyl) cyclobutane and/or r-1, t-2, c-3, t-4, c-5, t-6-hexa (methoxycarbonyl) cyclohexane.

Alternatively, an oxacarbocyclic oligomer of formula (IVb) is used:

wherein R is_cAnd R_dAs defined above, and n is an integer from 2 to 10 inclusive, more preferably from 2 to 6 inclusive.

The fumaric acid derivatives used according to the invention can be prepared according to known processes, as described in DE 19721099.6, DE 10133004.9 or DE 10217314.1. The contents of these publications are incorporated herein by reference.

The pharmaceutical formulation may be in a form suitable for oral, rectal, transdermal, dermal, ocular, nasal, pulmonary or parenteral administration. Preferably, the pharmaceutical formulation is suitable for oral administration. It may then exist in the following form: tablets, coated tablets, capsules, granules, solutions for drinking, liposomes, nanoparticles, nanocapsules, microcapsules, microtablets, pellets or powders, and may also be present in the form of: granules filled in capsules or sachets, microtablets filled in capsules or sachets, pellets filled in capsules or sachets, nanoparticles filled in capsules or sachets, or powders filled in capsules or sachets. Preferably, the drug is in the form of nanoparticles, pellets or minitablets, which may optionally be enclosed in sachets or capsules.

Preferably, all solid oral dosage forms may be provided with an enteric coating. For example, the enteric coating may be applied to tablets, microtablets, pellets, and the like, but may also be applied to capsules containing them.

Basically, the oral pharmaceutical forms according to the invention can be prepared according to classical compression methods as well as by direct compression, and can be made as solid dispersions according to the melt process or by spray-drying methods. If desired, the enteric coating can be poured or sprayed stepwise onto the core tablet in a classical coating pan, or applied by a fluidized bed apparatus according to known methods. Subsequently, after drying has been completed, a film coating may be applied in the same device.

Preferably, the fumaric acid derivatives used for the preparation of the pharmaceutical preparation according to the invention are used in such an amount that the pharmaceutical preparation contains one or more fumaric acid derivatives in an amount corresponding to and/or equal to an amount of 1-500mg, preferably 10-300mg, most preferably 10-200mg of fumaric acid per dosage unit.

In the case of parenteral administration by injection (iv, im, sc, ip), the formulations are in a form suitable for this administration. All conventional liquid carriers suitable for injection may be used.

According to a preferred embodiment, the medicaments to be produced according to the invention may contain the following components, alone or in admixture: 10-500mg of dialkyl fumarate, in particular dimethyl fumarate and/or diethyl fumarate, 10-500mg of alkyl fumarate calcium salt, in particular methyl fumarate calcium salt and/or ethyl fumarate calcium salt, 0-250mg of alkyl fumarate zinc salt, in particular methyl fumarate zinc salt and/or ethyl fumarate zinc salt, 0-250mg of hydrogen alkyl fumarate, in particular hydrogen methyl fumarate and/or hydrogen ethyl fumarate, and 0-250mg of alkyl fumarate magnesium salt, in particular methyl fumarate magnesium salt and/or ethyl fumarate magnesium salt, the sum of said amounts corresponding to 1-500mg, preferably 10-300mg and most preferably 10-200mg of fumaric acid equivalents (equivalent).

Particularly preferably, the formulations according to the invention used contain only 10 to 300mg of dimethyl fumarate.

According to a particularly preferred embodiment, the pharmaceutical preparation is in the form of a minitablet or a pellet. Preferably, they have a size and/or mean diameter of < 5000 microns, preferably of 300-. Due to the administration of the fumaric acid derivative in the form of the microtablets preferred according to the invention, gastrointestinal irritation and/or side effects which are unavoidable with conventional single unit dose tablets can be further reduced. This is presumably due to the fact that the microtablets, preferably enteric coated microtablets, have been distributed in the stomach and thus enter the intestine block by block (bolus) where the active substance is released in a locally smaller dose, while the total dose is the same. For this reason, local irritation of intestinal epithelial cells can be avoided, thereby resulting in better gastrointestinal tolerance of the microtablets than conventional tablets.

Preparation example

In order to explain the use according to the invention, a number of examples for the preparation of preferred pharmaceutical preparations are given below. The examples are for illustrative purposes only and do not limit the invention.

Example 1

Film-coated tablets with an enteric coating were prepared, containing 100.0mg of monomethyl fumarate calcium salt, corresponding to 78mg of fumaric acid

Taking necessary precautions (breathing mask, gloves, protective clothing, etc.), 10kg of monomethyl fumarate calcium salt are comminuted, mixed vigorously and homogenized through a 800 mesh sieve. Then, an excipient mixture was prepared which consisted of: 21kg of starch derivative (STA-RX: 1500. ANG.), 2kg of microcrystalline cellulose (Avicel PH 101. ANG.), 0.6kg of polyvinylpyrrolidone (PVP, Kollidon. RTM. 25), 4kg of Primogel. ANG., 0.3kg of colloidal silicic acid (Aerosil. ANG.).

The active ingredient is added to the entire powder mixture, mixed, homogenized with a 200 mesh sieve, then processed with a 2% aqueous solution of polyvinylpyrrolidone (Kollidon. RTM. 25) in the usual way to form binder granules, which are then mixed with the outer phase in the dry state. The outer phase consists of 2kg of a so-called FST compound, which contains 80% talc, 10% silicic acid and 10% magnesium stearate.

The mixture was then compressed by conventional means into tabs of 400mg weight and 10.0mm diameter. In addition to the typical compression method, other methods may be used to prepare tablets, such as direct compression or solid dispersion according to a melting method and a spray drying method.

Enteric coating:

2.250kg of a solution of hydroxypropylmethylcellulose phthalate (HPMCP, Pharmacoat HP-50) are dissolved in a solvent mixture consisting of 2.50 l of deionized water, 13 l of acetone Ph.Helv.VII and 13 l of ethanol (94% by weight), and 0.240kg of castor oil (Ph.Bur.II) is then added to the solution. The solution is poured or sprayed stepwise onto the core of the tablet in the coating pan in a conventional manner.

After drying accordingly, film coating is subsequently carried out. The coating (solution) consisted of Eudragit E12.5% 4.8kg, talc Ph.Eur.II 0.34kg, titanium oxide (VI) Cronus RN 56-0.52 kg, pigmented lacquer (colorated lacquer) ZLT-2 blue (Siegle)0.21kg, and polyethylene glycol 6000Ph.Helv.VII 0.12kg in a solvent mixture consisting of 8.2kg 2-propanol Ph.Helv.VII, 0.06kg Triacetin (Triacetin. RTM.) and 0.2kg deionized water. After being uniformly distributed in a coating pan or a fluidized bed, the coating is dried and polished according to a conventional method.

Example 2

Enteric-coated capsules containing 86.5mg of monoethyl calcium fumarate and 110.0mg of dimethyl fumarate, corresponding to a total of 150mg of fumaric acid, were prepared

Taking the necessary precautions (breathing mask, gloves, protective clothing, etc.), 8.65kg of calcium monoethyl fumarate and 11kg of dimethyl fumarate were mixed vigorously with a mixture consisting of 15kg of starch, 6kg of lactose Ph.Helv.VII, 2kg of microcrystalline cellulose (Avicel. RTM.), 1kg of polyvinylpyrrolidone (Kollidon. RTM. 25) and 4kg of Primogel. RTM. and homogenized through an 800 mesh screen.

The entire powder mixture is processed into bonded particles in the usual manner together with a 2% aqueous solution of polyvinylpyrrolidone (Kollidon. RTM. 25) and mixed with the outer phase in the dry state. The outer phase consists of 0.35kg of colloidal silicic acid (Aerosil), 0.5kg of magnesium stearate and 1.5kg of talc Ph.Helv.VII. This homogeneous mixture is filled in suitable capsules, 500.0mg each, which are then provided with an enteric (gastric resistant) coating consisting of hydroxypropyl ethylcellulose phthalate and castor oil as softener, according to known methods.

Example 3

Enteric microtablets ("tablet" for) in capsules were prepared containing 87.0mg of calcium monoethyl fumarate, 120mg of dimethyl fumarate, 5.0mg of magnesium monoethyl fumarate and 3.0mg of zinc monoethyl fumarate, which corresponded to a total of 164mg of fumaric acid

Taking necessary precautions (breathing mask, gloves, protective clothing, etc.), 8.7kg of calcium monoethyl fumarate, 12kg of dimethyl fumarate, 0.5kg of magnesium monoethyl fumarate and 0.3kg of zinc monoethyl fumarate were crushed, mixed vigorously and homogenized with a 800 mesh sieve. An excipient mixture having the following composition was prepared: 18kg of starch derivative (STA-RX 1500), 0.3kg of microcrystalline cellulose (Avicel PH 101), 0.75kg of PVP (Kollidon 120), 4kg of Primogel, 0.25kg of colloidal silicic acid (Aerosil). All powder mixtures were added to the active ingredient mixture, homogenized with a 200 mesh sieve, treated in the conventional manner with a 2% aqueous solution of polyvinylpyrrolidone (Kollidon K25) to give binder granules, and mixed in the dry state with an external phase consisting of 0.5kg of magnesium stearate and 1.5kg of talc. The powder mixture was pressed into tabs with a total mass of 10.0mg and a diameter of 2.0mm by conventional methods.

Enteric (gastric acid resistant) coatings were applied in a fluid bed apparatus. To achieve resistance to gastric acid, a solution of 2.250kg of hydroxypropylmethylcellulose phthalate (HPMCP, Pharmacoat HP50) was dissolved in a solvent mixture consisting of 13 liters of acetone, 13.5 liters of 94% by weight ethanol which had been denatured with 2% ketone and 2.5 liters of deionized water. To the solution obtained above, 0.240kg of castor oil was added as a softener, and then coated portion by portion on the tablet core according to a conventional method.

Film coating: after completion of the drying, a suspension having the following composition was applied as a film coating in the same apparatus: 0.340kg of talc, 0.2kg of titanium (VI) oxide Cronus RN 560.4kg, 868370.324kg of pigmented paint L red paint, 12.5% of Eudragit E4.8 kg, and 0.12kg of polyethylene glycol 6000pH 11XI in a solvent mixture consisting of 8.17kg of 2-propanol, 0.2kg of deionized water and 0.6kg of glyceryl triacetate (Triacetin).

The stomach acid resistant microtablets were analyzed for their composition and then filled in hard gelatin capsules at the corresponding dry weight and sealed.

Example 4

Enteric microtablets prepared in capsules containing 120.0mg dimethyl fumarate, which corresponds to 96mg fumaric acid

Taking necessary precautionary measures (breathing mask, gloves, protective clothing, etc.), pulverizing 12kg dimethyl fumarate, and homogenizing with 800 mesh sieve. An excipient mixture having the following composition was prepared: 17.5kg of starch derivative (STA-RX-1500), 0.30kg of microcrystalline cellulose (Avicel-PH 101), 0.75kg of PVP (Kollidon-120), 4kg of Primogel, 0.25kg of colloidal silicic acid (Aerosil). The entire powder mixture is added to the active ingredient mixture, mixed and homogenized with a 200-mesh sieve, processed into cohesive granules in a conventional manner with a 2% aqueous solution of polyvinylpyrrolidone (Kollidon _ K25), and mixed in the dry state with an external phase consisting of 0.5kg of magnesium stearate and 1.5kg of talc.

The powder mixture was then compressed into 2.0mm diameter convex microtablets having a total mass of 10.0mg by conventional methods.

To achieve resistance to gastric acid, 2.25kg of a hydroxypropylmethylcellulose phthalate (HPMCP, Pharmacoat HP-50) solution are dissolved in portions, for example in a solvent mixture consisting of 13 liters of acetone, 13.5 liters of ethanol (94% by weight, denatured with 2% of ketone) and 1.5 liters of deionized water. Castor oil (0.24kg) was added to the resulting solution as a softener and then coated stepwise onto the tablet cores according to conventional methods.

After completion of the drying, a suspension having the following composition was applied as a film coating in the same apparatus: 0.34kg of talc, 0.2kg of titanium oxide (VI), CronsuRN 560.4kg, tinctorial L red paint 868370.324kg, Eudragit E12.5% 4.8kg, and polyethylene glycol 6000pH 11XI 0.12kg in a solvent mixture consisting of 8.17kg of 2-propanol, 0.2kg of deionized water, and 0.6kg of glyceryl triacetate (Triacetin).

The stomach acid resistant microtablets were analyzed for their ingredients and then filled in hard gelatin capsules at the corresponding net weights and sealed.

Example 5

Enteric microtablets prepared in capsules containing 120.0mg of fumaric acid diglycine diamide, which corresponds to 96mg of fumaric acid

12kg of fumaric acid diglycine diamide were comminuted and homogenized as described above. An excipient mixture was prepared consisting of: 23.2kg microcrystalline cellulose (Avicel _ PH 200), 3kg sodium carboxymethylcellulose (AC-Di-SOL-SD-711), 2.5kg talc, 0.1kg anhydrous silicic acid (Aerosil _200) and 1kg magnesium stearate. The entire powder mixture is added to the active ingredient mixture and mixed homogeneously. The powder mixture was then compressed by direct compression into convex microtablets with a total mass of 10.0mg and a diameter of 2.0 mm. 2.0mm convex microtablets.

Subsequently, 0.94kg of Eudragit was preparedSolution in isopropanol, which additionally contains 0.07kg of dibutyl phthalate. The solution was sprayed onto the tablet cores. Then, 17.32kg of Eudragit was preparedLD-55 and a dispersion of a mixture of 2.8kg of micronized talc (micro-talcum), 2kg of Macrogol 6000 and 0.07kg of Dimeticon in water were sprayed onto the tablet cores.

Subsequently, the enteric minitablets were analyzed for their ingredients, filled in hard gelatin capsules at the corresponding dry weight and sealed.

Example 6

Enteric microtablets prepared in capsules containing 60.0mg r-1, t-2, c-3, t-4-tetrakis (methoxycarbonyl) cyclobutane and 30.0mg r-1, t-2, c-3, t-4, c-5, t-6-hexa (methoxycarbonyl) cyclohexane

6.0kg of r-1, t-2, c-3, t-4-tetrakis (methoxycarbonyl) cyclobutane and 3.0kg of r-1, t-2, c-3, t-4, c-5, t-6-hexa (methoxycarbonyl) cyclohexane were crushed, mixed vigorously and homogenized by passing through a 800 mesh sieve. Preparation of an excipient mixture having the following compositionCompound (a): 18kg of starch derivative (STA-RX 1500)) 0.30kg microcrystalline cellulose (Avicel PH 101), 0.75kg PVP (Kollidon 120), 4.00kg Primogel, 0.25kg colloidal silicic acid (Aerosil). The active ingredient is added to the entire powder mixture, homogenized by means of a 200-mesh sieve, processed in the customary manner with a 2% aqueous solution of polyvinylpyrrolidone (Kollidon K25) to give cohesive granules, and mixed with the outer phase in the dry state. The external phase consisted of 0.50kg magnesium stearate and 1.50kg talc. Thereafter, the powder mixture was compressed by conventional methods into convex microtablets having a total mass of 10.0mg and a diameter of 2.0 mm.

The enteric (gastric acid resistant) coating is poured onto the tablet cores in a typical coating pan. To achieve resistance to gastric acid, a solution of 2.250kg of hydroxypropylmethylcellulose phthalate (HPMCP, Pharmacoat HP50) was dissolved in portions in a solvent mixture consisting of 13.00 liters of acetone, 13.5 liters of 94% by weight ethanol which had been denatured with 2% ketone, and 2.50 liters of deionized water. To the resulting solution 0.240kg of castor oil was added as a softener and then coated portion by portion on the tablet core according to a conventional method.

Film coating: after the drying was completed, a suspension of the following composition was applied as a film coating in the same apparatus: 0.340kg talc, 0.200kg titanium oxide (VI) Cronus RN 560.400kg, 868370.324kg pigmented paint L red paint, Eudragit E12.5% 4.800kg, and polyethylene glycol 6000pH 11XI 0.120kg in a solvent mixture consisting of 8.170kg 2-propanol, 0.200kg deionized water, and 0.600kg glyceryl triacetate (Triacetin).

Subsequently, the stomach acid resistant microtablets were analyzed for their composition and then filled in hard gelatin capsules at the corresponding dry weight and sealed.

Example 7

A suspension for parenteral administration is prepared containing 60.0mg of r-1, t-2, c-3, t-4-tetrakis (methoxycarbonyl) cyclobutane and 30.0mg of r-1, t-2, c-3, t-4, c-5, t-6-hexa (methoxycarbonyl) cyclohexane

Ingredient mg/ml

r-1, t-2, c-3, t-4-tetra (methoxycarbonyl) cyclobutane 60

r-1, t-2, c-3, t-4, c-5, t-6-hexa (methoxycarbonyl) cyclohexane 30

Methyl cellulose 0.25

Sodium citrate dihydrate 30

Benzyl alcohol 9.00

Methylparaben 1.8

Propyl p-hydroxybenzoate 1.2

Water for injection q.s.a.d.1.00

The foregoing ingredients are processed into parenteral suspensions using standard techniques.

Application example

Example A

In vivo data of DMF treatment of cardiac insufficiency using rat model.

The effect of dimethyl fumarate was tested in this experiment using an acute ischemia and reperfusion model in rats. For this purpose, healthy female rats were divided into 3 groups of 17 rats each. In the experiment, ischemia was caused by occlusion of arteries while exposing the heart for 45 minutes, followed by reperfusion for 120 minutes. Finally, myocardial infarction was caused by re-occlusion and the area of risk was determined by phthalocyanine blue staining.

At the beginning of the first occlusion, the test substance is administered intravenously. The control group received 0.02% DMSO (0.5ml/kg body weight) and the DMF group received 10mg dimethyl fumarate (0.5ml/kg body weight) in 0.02% DMSO. In group 2, animals were subjected to ischemic preconditioning (ischemia and reperfusion, respectively, twice for 5 minutes).

The results are shown in FIG. 1. Clearly, dimethyl fumarate (DMF) and Ischemic Preconditioning (IPC) limited the size of the infarct statistically significantly in our experiments, with the risk zones being similar in all 3 groups. Thus, the data demonstrate that dimethyl fumarate is used to significantly reduce infarct size and thereby prevent cardiac insufficiency.

Example B

Inhibition of PDGF-induced thymidine incorporation

Successful treatment of asthma involves 3 different pathways: (1) reducing the release of inflammatory mediators in allergy, (2) inhibiting T-lymphocyte invasion, and (3) inhibiting mesenchymal cell proliferation. Glucocorticoids, which are the treatment of choice in asthma, have been shown to inhibit mesenchymal cell proliferation. Thus, this experiment can be used to screen for possible other active substances for the treatment of asthma.

In the presence and absence of 10^-5BSM (bronchial smooth muscle) was cultured in RPMI, 0.3% albumin and 0.1% DMSO at 37 ℃ in the presence of 0, 1, 5, 10 and 20ng/ml PDGF in the presence of M dimethyl fumarate.

After a predetermined time, 5 μ Ci³H-thymidine was added to the medium and incubation continued for 24 hours. Finally, the incorporation was terminated by centrifugation, the supernatant was removed, the cells were washed and lysed. Determination of radioactivity in lysate by comparison to control in liquid scintillation device³Incorporation of H-thymidine. The results are shown in FIG. 2, and are expressed as a comparisonPercentage value of group (100%) comparison. The addition of PDGF clearly enhances³Incorporation of H-thymidine and thus enhanced cell proliferation, whereas the enhancement was significantly reduced upon addition of dimethyl fumarate.

Example C

Bronchial smooth muscle cells were grown in 96-well plates until they reached 60-70% confluence. Cells were then starved for 48 hours in serum-free RPMI medium containing 0.3% albumin. 1 hour before stimulation of cell proliferation with 10ng/ml PDGF, (a)10^-5M DMF，(b)10^-8M dexamethasone (dexa), or (c)10^-5Mdmf and 10^-8M dexa treated cells. Untreated cells (buffer only) were used as controls. Cells were treated for 36 hours, after which 4uCi of³H-thymidine was added for another 8 hours. Lysing the cells to incorporate DNA³H-thymidine was bound to the filter and the cpm of incorporation was measured in a liquid scintillation device. The results are shown in figure 3, expressed as a percentage of the control group (100%) and compared to PDGF-induced proliferation.

When only the therapeutically correct dose of dexa (10) is used^-8M) reduced cell proliferation to about 116. + -.11% when the cells were treated. In use 10^-5Similar reductions (117 ± 4%) were observed upon DMF treatment of M. The combined administration of these concentrations of DMF and dexa synergistically reduced cell proliferation to near baseline levels (95 ± 11%). These results indicate that DMF can be used alone or often in combination with dexamethasone or glucocorticoids to treat asthma.

In a particularly preferred embodiment for the treatment of asthma and chronic obstructive pulmonary disease, the treatment is thus in combination with a glucocorticoid. The administration can be in the same dosage unit or in separate dosage units. Administration may also be in parallel or sequentially. Preferably, the glucocorticoid is selected from the group consisting of: dexamethasone, cortisone, hydrocortisone, prednisolone, prednisone, methylprednisolone, fluocortolone, triamcinolone, betamethasone, beclomethasone, budesonide, flunisolide, fluticasone, and pharmaceutically acceptable salts and derivatives thereof. Most preferably, the glucocorticoid is dexamethasone.

Example D

Different doses of DMF were administered daily to salt-sensitive Dahl rats and were fed a high-salt meal. After 8 weeks of treatment, the end-stage left ventricular dilatation diameters were determined by echocardiographic analysis for the test and control groups. The measured group was a control group (0mg DMF; n ═ 9); group 1(2 × 5mg DMF/kg/d; n ═ 9) and group 2(2 × 15mg DMF/kg/d; n ═ 11).

In echocardiography analysis, DMF prevented left ventricular dilation after 8 weeks of high salt meals in a dose-dependent manner. Specifically, the inner diameter of the left ventricle in the DMF group remained within the same range as baseline (see fig. 4). In contrast, animals in the control group exhibited enlarged left ventricles, indicating left ventricular dilation. Importantly, left ventricular dilation marks the transition from compensatory hypertrophy to decompensated heart failure. Therefore, DMF delays the transition to heart failure and thus prevents myocardial infarction.

Claims (21)

1. Use of a fumaric acid derivative for the production of a medicament for the treatment of asthma and chronic obstructive pulmonary disease, wherein the fumaric acid derivative is selected from one or more dialkyl fumarates of formula I

Wherein R is₁And R₂May be the same or different and independently represent a linear, branched or cyclic,Saturated or unsaturated C_1-24An alkyl group, a carboxyl group,

or one or more monoalkyl fumarates of formula II

Wherein

R₁Represents linear, branched or cyclic, saturated or unsaturated C_1-24An alkyl group, a carboxyl group,

a represents hydrogen, an alkali metal or alkaline earth metal cation or a physiologically acceptable transition metal cation, and

n is equal to 1 or 2 and corresponds to the valence of A.

2. Use according to claim 1, wherein A is selected from Li⁺、Na⁺、K⁺、Mg²⁺、Ca²⁺、Zn²⁺、Fe²⁺And Mn²⁺。

3. Use according to claim 1, wherein the asthma is asthma caused by allergy, infection, analgesics, working conditions or exertion, or cardiac asthma.

4. Use according to claim 1 in combination with a glucocorticoid.

5. Use according to claim 4, wherein the glucocorticoid is selected from the group consisting of: dexamethasone, cortisone, hydrocortisone, prednisolone, prednisone, methylprednisolone, flucolone, triamcinolone, beclomethasone, budesonide, flunisolide, fluticasone, betamethasone, and pharmaceutically acceptable salts thereof.

6. Use according to claim 1, wherein the fumaric acid derivative is selected from one or more compounds of formulae (I) and (II) and mixtures thereof.

7. Use according to claim 1, wherein the fumaric acid derivative is selected from the group consisting of: dimethyl fumarate, diethyl fumarate, ethyl methyl fumarate, methyl hydrogen fumarate, ethyl hydrogen fumarate, calcium methyl fumarate, calcium ethyl fumarate, magnesium methyl fumarate, magnesium ethyl fumarate, zinc methyl fumarate, zinc ethyl fumarate, iron methyl fumarate, iron ethyl fumarate, and mixtures thereof.

8. Use according to claim 1, wherein the alkyl group having 1 to 24 carbon atoms is selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, cyclopentyl, 2-ethylhexyl, hexyl, cyclohexyl, heptyl, cycloheptyl, octyl, vinyl or allyl.

9. Use according to claim 8, wherein the alkyl group having 1 to 24 carbon atoms is a methyl or ethyl group.

10. Use according to claim 1, wherein the medicament is provided in a form suitable for oral, rectal, dermal, ocular, nasal or pulmonary administration.

11. The use according to claim 1, wherein the medicament is provided in a form suitable for parenteral administration.

12. Use according to claim 10 or 11, wherein the medicament is provided in the form of: tablet, capsule, granule, microcapsule or powder.

13. Use according to claim 10 or 11, wherein the medicament is provided in the form of: coated tablets, granules filled in capsules, microtablets filled in capsules, pellets filled in capsules, nanoparticles filled in capsules, or powders filled in capsules.

14. Use according to claim 10 or 11, wherein the medicament is in the form of nanocapsules, pellets or microtablets.

15. Use according to claim 12, wherein the dosage form is provided with an enteric coating.

16. Use according to claim 13, wherein the dosage form is provided with an enteric coating.

17. Use according to claim 14, wherein the dosage form is provided with an enteric coating.

18. Use according to any one of claims 1 to 11, wherein the amount of fumaric acid derivative in the medicament corresponds to 1-500mg of fumaric acid.

19. An in vitro method for inhibiting proliferation of bronchial smooth muscle cells, comprising the step of contacting bronchial smooth muscle cells, directly or indirectly, with a proliferation-inhibiting amount of a fumaric acid derivative, wherein said fumaric acid derivative is selected from one or more dialkyl fumarates of formula I

Wherein R is₁And R₂May be the same or different and independently represent linear, branched or cyclic, saturated or unsaturated C_1-24An alkyl group, a carboxyl group,

or one or more monoalkyl fumarates of formula II

Wherein

R₁Represents linear, branched or cyclic, saturated or unsaturated C_1-24An alkyl group, a carboxyl group,

a represents hydrogen, an alkali metal or alkaline earth metal cation or a physiologically acceptable transition metal cation, and

n is equal to 1 or 2 and corresponds to the valence of A.

20. The method according to claim 19, wherein a is selected from Li⁺、Na⁺、K⁺、Mg²⁺、Ca²⁺、Zn²⁺、Fe²⁺And Mn²⁺。

21. The process according to claim 19 or 20, wherein the fumaric acid derivative is selected from one or more compounds of formulae (I) and (II) and mixtures thereof.