HRP20010429A2 - Formulation with an improved therapeutic range, containing nucleotide synthesis inhibitors - Google Patents

Description

Highly basic anion exchangers are used as lipid-lowering agents in heterozygous familial hypercholesterolemia and other primary hyperlipoproteinemias with a particular increase in LDL-fractions, or collagenous diarrhea. Examples of suitable active substances that can be used as lipid-lowering agents are N-(2-aminoethyl)-N'-[2-[(2-aminoethyl)amino]-ethyl]-1,2-ethanediamine polymer with (chloromethyl) -oxirane, which is called colestipol (Colestid®) or cholestyramine (CAS-No. 11 041-12-6), which is a styrenedivinyl-benzyl copolymer. Derivatives of isoxazole or crotonic acid amide are described in patent applications EP 484 223; EP 529 500; US 4,061,767; EP 538 783 or EP 551 230. Compounds, which inhibit the synthesis of purines or pyrimidines, and are designated as inhibitors of nucleotide synthesis (Burkhardt and Kalden; Rheumat. Int. (1997); 17:85-90), for example compounds of the formula I and/or II, brekinar (6-fluoro-2-(2'-fluoro[1,1'-biphenyl]-4-yl)-3-methyl-4-quinolinecarboxylic acid), mycophenolate mofetil (2-morpholinoethyl-( E)-6-(1,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxoisobenzofuran-5-yl)-4-methyl-4-hexenoate), methotrexate (CAS-No. 59- 05-02) or mizoribine (CAS-No. 50924-49-7), after oral application are resorbed in the intestines of the patient and soon after a short-term increase in the amount in the blood, after intake (resorption peak value) lead to a constant high level in the blood. The above-mentioned inhibitors of nucleotide synthesis are re-excreted via the liver and bile into the intestines. The mentioned, excreted compounds from the intestine can be partially reabsorbed and enter the blood. The mentioned compounds are therefore prone to enterohepatic circulation.

When using inhibitors of nucleotide synthesis to influence the immune system, it was surprisingly found that only a short-term effect of these substances is sufficient for the desired effect on the immune system. If the amount of these substances that leads to an effective effect is properly maintained in the blood over a long period of time, then the side effects decrease, but the desired effect on the immune system does not increase. By limiting the effect to a short period of time, the tolerability of the therapy can be improved while properly maintaining the desired pharmacodynamic effect on the immune system (= improving the therapeutic range).

In the case of inhibitors of nucleotide synthesis that are prone to enterohepatic circulation, the duration of action can be shortened by administering substances that interrupt enterohepatic circulation. Interruption of the enterohepatic circulation maintains the desired effect on the immune system, however side effects are significantly reduced. The above-mentioned inhibitors of nucleotide synthesis may also show an improved therapeutic breadth of action if the action of the compounds that antagonize the action of the inhibitors of nucleotide synthesis is time-delayed - so that they are administered later than the inhibitors of nucleotide synthesis.

In addition, the term therapeutic range refers to the measure of tolerability of the drug, and it is mainly the range between the lowest dose that still leads to the desired therapeutic effect and the dose that leads to side effects. The measure of the achieved improvements are, for example, the amount of red blood cells, hemoglobin content, hematocrit, amount of glutamate-oxalacetate-transaminase, glutamate-pyruvate-transaminase, alkaline phosphatase (from bone marrow) or amylase and weight compared to untreated patients. The invention relates to a preparation, which contains

1) at least one compound that substantially prevents the enterohepatic circulation of nucleotide synthesis inhibitors or temporarily delays the action of nucleotide synthesis inhibitors, and

2) at least one nucleotide synthesis inhibitor from the group consisting of brekinar, mycophenolate mofetil, 2-morpholinoethyl-(E)-6-(1,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxoisobenzofuran-5- yl)-4-methyl-4-hexenoate, methotrexate, mizoribine and compounds of formula I or II

[image]

and/or, if necessary, a stereoisomeric form of the compound of formula I or II and/or a physiologically tolerable salt of the compound of formula II, in which

R1 represents

a) (C1-C4)-alkyl,

b) (C3-C5)-cycloalkyl,

c) (C2-C6)-alkenyl or

d) (C2-C6)-alkynyl,

R2 is

a) -CF3,

b) -O-CF3,

c) -S-CF3,

d) -OH,

e) -NO2,

f) halogen,

g) benzyl,

h) phenyl,

i) -O-phenyl,

k) -CN or

l) -O-phenyl, singly or multiply substituted with

1) (C1-C4)-alkyl,

2) halogen, or

3) -O-CF3 or

4) -O-CH3,

R3 is

a) (C1-C4)-alkyl,

b) halogen, or

c) hydrogen atom, i

X is

a) -CH-group or

b) nitrogen atom.

A mixture that inhibits the synthesis of nucleotides and compounds of formula I and II or salts of compounds of formula II and a mixture of compounds that substantially prevent the enterohepatic circulation of compounds of formula I or II can be used.

By the term "compound, which significantly prevents the enterohepatic circulation of the compound of formula I or II" is meant, for example, strongly basic anion exchangers such as colestipol and cholestyramine or activated charcoal. By the term "compounds, which by delaying it, antagonize the action of inhibitors of nucleotide synthesis" are meant compounds such as uridine, purine, purine nucleotides or pyrimidine nucleotides.

It is preferable to use the compound of formula I and/or II and/or, if necessary, the stereoisomeric form of the compound of formula I or II and/or the salt of the compound of formula II, in which formula

R1 is

a) methyl,

b) cyclopropyl or

c) (C3-C5)-alkyl,

R2 is -CF3 or -CN,

R 3 is a hydrogen atom or methyl, and

X is a -CH group, in combination with at least one compound from the group consisting of colestipol, cholestyramine and activated carbon.

The use of N-(4-trifluoromethyl-phenyl)-5-methylisoxazole-4-carboxamide, N-(4-trifluoromethyl-phenyl)-2-cyano-3-hydroxy-crotonic acid amide, 2-cyano-3- cyclopropyl-3-hydroxy-acrylic acid-(4-cyano-phenyl)-amide or N-(4-trifluoromethylphenyl)-2-cyano-3-hydroxy-hept-2-ene-6-yne-carboxylic acid-amide in in combination with cholestyramine.

The production of compounds of formula I or II is carried out by a known process described in EP 484 223; EP 529 500; US 4,061,767; EP 538 783 or in EP 551 230. The starting substances for the chemical conversion are known or can be easily produced by methods known from the literature.

The term alkyl, alkenyl or alkynyl refers to residues whose carbon chain can be straight or branched. Furthermore, alkenyl or alkynyl residues may also contain several double or triple bonds. Cyclic alkyl radicals are, for example, 3- to 5-membered monocycles such as cyclopropyl, cyclobutyl or cyclopentyl. Salts of the compound of formula II are, for example, sodium salts or salts of lysine, which can be produced in the manner described in European patent application no. EP 0769296.

The preparation according to the invention is suitable, for example, for treatment

- immunological diseases,

- inflammatory and cytotoxic processes associated with gene therapy procedures,

- cancer treatment, such as lung cancer, leukemia, ovarian cancer, sarcoma and Kaposi's sarcoma, meningioma, bowel cancer, lymph node cancer, brain tumors, breast cancer, pancreatic cancer, prostate cancer or skin cancer,

- autoimmune diseases such as systemic lupus erythematosus or multiple sclerosis,

- rheumatic diseases,

- in transplants or host-against-graft reactions or graft-against-host reactions,

- diseases caused by strong cell proliferation,

- psoriasis or atypical dermatitis,

- allergies, asthma, urticaria, rhinitis or uveitis,

- type II diabetes,

- cystic fibrosis, colitis, liver fibrosis or sepsis,

- chronic inflammatory diseases such as arteriosclerosis, Crohn's disease or ulcerative colitis.

The invention also relates to a method for the production of a preparation, which is indicated by the fact that the inhibitor of nucleotide synthesis and a compound that significantly prevents the enterohepatic circulation of the inhibitor of nucleotide synthesis, or, by increasing it over time, antagonizes the action of the inhibitor of nucleotide synthesis, is processed into a form suitable for application together with a pharmaceutically suitable and physiologically tolerable carrier and, if necessary, further suitable active, additional or auxiliary substances.

The preparation according to the invention can also contain a composition or combined packages in which the ingredients are placed next to each other and therefore can be administered simultaneously, separately or gradually in time in one and the same human or animal body. Time-graded application of the compound of formula I and/or application of a compound that substantially prevents the enterohepatic circulation of the compound of formula I or II is preferred. For this purpose, for example, N-(4-trifluoromethylphenyl)-2-cyano-3-hydroxy-crotonic acid amide (referred to below as compound 1) is first applied. Cholestyramine, which significantly prevents the enterohepatic circulation of compound 1, is applied later, i.e. 2 or 4 hours after the application of compound 1. With this time-shifted administration of compound 1 and cholestyramine, compound 1 is first resorbed unhindered from the digestive tract. After application of cholestyramine, which is not resorbed systemically, compound 1, excreted via the bile, binds to cholestyramine and therefore cannot be reabsorbed; this results in interruption of the enterohepatic circulation. In this way, the duration of action and the amount of compound 1 in the blood are significantly reduced. Despite this significantly reduced amount in the blood, the efficacy in a pathological animal model, such as adjuvant-induced arthritis at lower, actually still effective dosages of approximately 2.5 mg/kg/day of compound 1, is not reduced by cholestyramine administration. When a high dosage of 25 mg/kg/day of compound 1, which already leads to various side effects, is applied in the same animal model, a clear reduction of side effects is observed when cholestyramine is administered while maintaining the desired effect on the immune system.

The preparation according to the invention can be made in the form of a drug dosage unit such as capsules (including microcapsules, which generally do not contain any pharmaceutical carriers), tablets including dragees and pills, or suppositories, whereby when using capsules, the capsule material can have the function of a carrier and it may contain content, for example, in the form of a powder, gel, solution, emulsion or dispersion. However, in any case, it is particularly advantageous to produce oral or peroral formulations with both active components 1) (e.g. cholestyramine) and 2) (compound of formula I and/or II), which contain calculated amounts of active substances together with the respective, desired pharmaceutical carrier . The appropriate formulation (suppositories) for rectal therapy can also be applied. Transdermal application in the form of ointment or cream, parenteral (intraperitoneal, subcutaneous, intramuscular) injection or oral application of a solution containing the combination according to the invention is also possible. In addition to the active substance, ointments, pastes, creams and powders may contain usual carriers, for example animal or vegetable fats, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, silicon dioxide, aluminum hydroxide, talc, zinc oxide, milk sugar, bentonite, calcium silicate and polyamide powder or a mixture of these substances. Tablets, pills or granules can be produced by processes such as pressing, dipping, swirling and making dragees in a boiler and they contain a carrier material and common excipients such as gelatin, agarose, starch (e.g. potato, corn or wheat starch), cellulose as ethyl cellulose, silicon dioxide, magnesium stearate, various sugars such as milk sugar and/or calcium phosphate. The solution for making dragees usually consists of sugar and/or thick syrup and usually also contains gelatin, synthetic cellulose ester, gum arabic, polyvinylpyrrolidone, pigments, surfactants, softeners and similar additives in accordance with the state of the art. Any conventional flow control, lubricating or sliding agent such as magnesium stearate and mold release agent can be used for the production of molded articles. Preference is given to preparations in the form of tablets with a shell and a core or multi-layered tablets, where the active component 2 is in the shell, i.e. in the core in one layer, while the active component 1 is in the core, in the shell or in another layer. Active components can also be in a delayed form or absorbed on a material for a delayed release, or included in a material for a delayed release (eg based on cellulose or polystyrene resin, eg hydroxyethyl cellulose). Slow release of the active substance can also be achieved by coating the layer or compartment in question with a conventional coating insoluble in gastric acid. It is preferable to slow down the release of the compound which significantly prevents the enterohepatic circulation of the compound of formula I or II.

The applied dosage depends, of course, on various factors, such as the species being treated (i.e. human or animal), age, weight, general state of health, degree of severity of the symptoms of the disease being treated, possible concomitant diseases (if present), method of concomitant treatment with other drugs or frequency of treatment. Dosing is generally given several times a day, preferably one to three times a day. The amounts of individual substances used are adjusted here according to the recommended daily dose of the active substance in question and generally in the combined preparation they should be present in an amount of 10% to 300% of the recommended daily dose, preferably from 50% to 150%, especially 80%. Suitable therapy with the combinations according to the invention therefore consists, for example, in the administration of one, two or three individual doses of a preparation consisting of N-(4-trifluoromethylphenyl)-5-methylisoxazole-4-carboxamide) or N-4-trifluoromethylphenyl)-2 -cyano-3-hydroxy-crotonxic acid-amide in an amount of 2 mg to 250 mg, preferably 5 mg to 150 mg, especially 10 mg to 50 mg, especially preferably 10 mg to 20 mg, and cholestyramine in an amount of 250 mg to 6000 mg , especially from 1500 mg to 3000 mg.

Furthermore, the preparations according to the invention can also be used together with other suitable active substances such as, for example, antiuricopatics, analgesics, steroidal or non-steroidal anti-inflammatory drugs, platelet aggregation inhibitors, cytokines, cytokine agonists, cytokine antagonists or immunosuppressive compounds such as cyclosporin A, FK 506 or rapamycin.

Example 1

Adjuvant-induced arthritis, modified by Perper (Proc. Soc. exp. Biol. Med. 137, 506 (1971))

Male Levis rats (Moellegard, Denmark) with a body weight of 160 to 210 g were used as experimental animals. On the first day, the animals received a subcutaneous injection in the root of the tail with complete Freund's Adjuvant, which contained a suspension of Mycobacterium butyricum in heavy paraffin oil (Difco; 6 mg/kg in paraffin oil; Merck). The compounds N-(4-trifluoromethylphenyl)-2-cyano-3-hydroxy-crotonic acid-amide and cholestyramine were suspended in carboxymethyl cellulose (1% in water) and administered orally. The compounds were applied once a day from the 1st to the 17th day of the experiment; then, on the 18th day, paw volume and arthritis index were determined.

The severity of the disease was determined by measuring paw volume on both hind paws. The measurement was performed using the water displacement method with a Plethysmometer 2060 (Rhema-Labortechnik, Hofheim, Germany). After that, the arthritis index was determined on the 18th day after the injection.

Determining the arthritis index:

1. ears 0.5 points for each ear where redness has appeared and where nodules have formed,

2. nose 1 point for swelling of the connective tissue,

3rd tail 1 point for submerging the knot,

4. front paws

0.5 points for each paw on which at least one joint inflammation appeared,

5. hind paws

1 point for mild inflammation (swelling),

2 points for moderate inflammation,

3 points for extensive inflammatory reaction.

Animals of one control group "arthritis control" received on day 1 one subcutaneous injection into the root of the tail of complete Freund's Adjuvant and received only the solvent (1% carboxymethylcellulose in water). Six animals were used for each dosage and in the control group. As a further control group, "healthy control", untreated animals were used. The criterion of effect was the reduction by half of the increase in the volume of the paw and the reduction of the arthritis index compared to the untreated control group and the weight of the animals in each case in percentages and in relation to the arthritis control. In the following table, N-(4-trifluoromethylphenyl)-2-cyano-3-hydroxy-crotonic acid-amide is designated as compound 1. Cholestyramine was applied 4 hours after compound 1. Table 1 shows the results obtained.

Table 1

[image]

Values listed in the table with the sign "-" represent data; all other values show an increase compared to the beginning of the experiment.

Animals that received the preparation according to the invention showed an increase in weight, which at the amount of 2.5 and 7.5 g of compound 1 was very close to the healthy control and significantly better than with compound 1 alone, while the effectiveness of compound 1 remained completely maintained.

Example 2

The experimental conditions are analogous to example 1. The effects of compound 1 and cholestyramine on the amount of red blood cells (RBC), hemoglobin content (HGB), hematocrit (HCT), on the amount of glutamate-oxalacetate-transaminase (CGOT) and glutamate-pyruvate-transaminase ( GPT). Cholestyramine was administered 4 hours after compound 1. Table 2 shows the results obtained.

Table 2

[image]

Values listed in the table with the sign "-" represent data; all other values show an increase compared to the beginning of the experiment.

Animals that received the preparation according to the invention showed a normal amount of red blood cells (RBC), hemoglobin content (HGB), hematocrit (HCT), amount of glutamate-oxalacetate-transaminase (GOT) and glutamate-pyruvate-transaminase (GPT), healthy control was very close and significantly better than in animals that received only compound 1, while the effect of compound 1 was completely retained.

Example 3

The experimental conditions are analogous to example 1. The effect of compound 1 and cholestyramine on the amount of alkaline phosphatase (AP) and amylase was determined. Cholestyramine was applied 4 hours after compound 1. Table 3 shows the results obtained.

Table 3

[image]

Values listed in the table with the sign "-" represent data; all other values show an increase compared to the beginning of the experiment.

Animals that received the preparation according to the invention showed a normalization of the amount of alkaline phosphatase, the healthy control was very close and significantly better than the group that received only compound 1, while the action of compound 1 remained completely preserved.

Example 4

The preparation according to the invention consisted of a small hard gelatin capsule containing 400 mg of cholestyramine and a larger hard gelatin capsule containing 20 mg of N-(4-trifluoromethyl-phenyl)-5-methylisoxazole-4-carboxamide. The smaller gelatin capsule was completely surrounded by the larger capsule. Glucose was used as a filling material between the two capsules.

Claims (8)

1. Preparation, characterized in that it contains 1) at least one compound from the group consisting of colestipol, cholesterin and activated carbon, which significantly prevents the enterohepatic circulation of nucleotide synthesis inhibitors or a compound from the group consisting of uridine, purine, purine nucleotides or pyrimidine nucleotides that antagonize the action of nucleotide synthesis inhibitors by delaying it , i 2) at least one inhibitor of nucleotide synthesis from the group consisting of 2-morpholinoethyl-(E)-6-(1,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxoisobenzofuran-5-yl)-4 -methyl-4-hexenoate, mizoribine and compounds of formulas I and II [image] and/or, if necessary, a stereoisomeric form of the compound of formula I or II and/or a physiologically tolerable salt of the compound of formula II, in which R1 represents a) (C1-C4)-alkyl, b) (C3-C5)-cycloalkyl, c) (C2-C6)-alkenyl or d) (C2-C6)-alkynyl, R2 is a) -CF3, b) -O-CF3, c) -S-CF3, d) -OH, e) -NO2, f) halogen, g) benzyl, h) phenyl, i) -O-phenyl, k) -CN or l) -O-phenyl, singly or multiply substituted 1) with (C1-C4)-alkyl, 2) halogen, 3) -O-CF3 or 4) -O-CH3, R3 is a) (C1-C4)-alkyl, b) halogen, or c) hydrogen atom, i X is a) -CH-group or c) nitrogen atom. 2. The preparation according to claim 1, characterized in that the compound of formula I and/or II and/or, if necessary, the stereoisomeric form of the compound of formula I or II and/or the salt of the compound of formula II is used, in which formula R1 is a) methyl, b) cyclopropyl or c) (C3-C5)-alkyl, R2 is -CF3 or -CN, R 3 is a hydrogen atom or methyl, and X is a -CH group. 3. Preparation according to claim 1 or 2, characterized in that N-(4-trifluoromethyl-phenyl)-5-methylisoxazole-4-carboxamide is used as the compound of formula I, or N-(4-trifluoromethylphenyl) is used as the compound of formula II )-2-cyano-3-hydroxycrotonamide, 2-cyano-3-cyclopropyl-3-hydroxy-acrylic acid-(4-cyanophenyl)amide or N-(4-trifluoro-methylphenyl)-2-cyano-3-hydroxyhept- 2-ene-6-yne-carboxamide. 4. Preparation according to one or more claims 1 to 3, characterized in that it additionally contains active substances from the group consisting of antiuricopatics, analgesics, steroidal or non-steroidal anti-inflammatory drugs, cytokines, cytokine agonists, platelet aggregation inhibitors, cytokine antagonists or immunosuppressive compounds such as cyclosporin A, FK 506 or rapamycin. 5. Preparation according to one or more claims 1 to 4, characterized in that it consists of formulations or combined packages in which the ingredients are placed next to each other and are therefore applied simultaneously, separately or sequentially on one and the same human or animal body. 6. The preparation according to claim 5, characterized by the fact that the application of the compound of formula I and/or II is carried out in time before the application of the compound that essentially prevents the enterohepatic circulation of the compound of formula I or II. 7. The use of a preparation according to one or more claims 1 to 6, characterized in that it is used for the production of a drug for the treatment of immunological diseases, inflammatory and cytotoxic processes associated with gene therapy procedures, carcinomatous disorders such as lung cancer, leukemia, ovarian cancer , sarcoma and Kaposi's sarcoma, meningioma, colon cancer, lymph node cancer, brain tumors, breast cancer, pancreatic cancer, prostate cancer or skin cancer, autoimmune diseases such as systemic lupus erythematosus or multiple sclerosis, rheumatic diseases, transplants or host reactions against graft or graft-versus-host reactions, diseases caused by severe cell proliferation, psoriasis or atypical dermatitis, allergies, asthma, urticaria, rhinitis or uveitis, and type II diabetes, cystic fibrosis, colitis, liver fibrosis or sepsis, chronic inflammatory diseases such as arteriosclerosis, Crohn's disease or ulcerative colitis. 8. The method for producing the preparation according to claims 1 to 8, characterized in that at least one inhibitor of nucleotide synthesis from the group consisting of 2-morpholinoethyl-(E)-6-(1,3-dihydro-4-hydroxy-6-methoxy -7-methyl-3-oxo-isobenzofuran-5-yl)-4-methyl-4-hexenoate, mizoribine and compounds of formula I or II, and a compound from the group consisting of colestipol, cholestyramine and activated charcoal, which significantly prevent enterohepatic circulation inhibitor of nucleotide synthesis, or a compound from the group consisting of uridine, pyrin, purine nucleotides or pyrimidine nucleotides which, by delaying it in time, antagonize the action of the inhibitor of nucleotide synthesis, processed into a form suitable for application together with a pharmaceutically suitable and physiologically tolerable carrier and, if necessary, with further suitable active and additional or auxiliary substances.