DE19814546A1 - (S) -2- (Biphenyl-4-sulfonylamino) -3- (1H-indol-3-yl) propionic acid derivatives - Google Patents

Abstract

The invention relates to (S)-2-(biphenyl-4-sulphonylamino)-3-(1H-indolyl)-propionic acid derivatives. S enantiomers of the compound of formula (1) are suitable for the production of medicaments used in the prophylaxis and therapy of diseases in the course of which matrix-catabolic metal proteinases are involved.

Description

The invention relates to new (S) - (biphenyl-4-sulfonylamino) -3- (1H-indol-3-yl) - propionic acids, process for their preparation and use thereof as Drug.

Special arylsulfonamino carboxylic acids serve as intermediates for Presentation of thrombin inhibitors (EP 0 468 231). In the registration EP 0 757 037 also describes the effect of sulfonylamino acid derivatives Metalloproteinase inhibitors are described. In the application WO 97/27174 Biphenyl-4-sulfonylamino-propionic acid derivatives and their effect as Metalloproteinase inhibitors described. In the German patent application DE 197 19 621.7 describes sulfonylaminocarboxylic acids via their However, bioavailability is not reported.

Furthermore, the arylsulfonyl group has proven to be an effective protecting group for the Proven amino function of α-aminocarboxylic acids (R. Roemmele, H. Rapoport, J. Org. Chem. 53 (1988) 2367-2371).

In the pursuit of effective compounds for the treatment of connective tissue to find diseases, it has now been found that the invention substituted sulfonylaminocarboxylic acids strong inhibitors of matrix Metalloproteinases are. The inhibition of stromelysin (matrix Metalloproteinase 3) and the neutrophil collagenase (MMP-8) special value since both enzymes break down the proteoglycans as important components of cartilage tissue, are significantly involved (A. J. Fosang et al. J. Clin. Invest. 98 (1996) 2292-2299). The compounds of the invention also show significantly improved pharmacokinetic properties after in vivo administration.  

Although the racemic mixture as well as the R and S enantiomers of the Compound of Formula I essentially similar in vitro inhibitory values to that Have matrix metalloproteinases stromelysin and neutrophil collagenase, surprisingly show only the S enantiomers compared to the R Enantiomers have up to a factor of 10 higher in vivo bioavailability and longer Residence times in the body of the animals examined.

The invention therefore relates to the (S) -enantiomer of the compound of the formula I.

and / or a physiologically acceptable salt of the S-enantiomer of the compound of formula I, wherein
R ^{1 represents} hydrogen atom, fluorine, chlorine or bromine and
R ^{2 represents} hydrogen atom, fluorine, chlorine, bromine or -OR ³ ,
wherein R ^{3 is} hydrogen, (C ₁ -C ₆ ) alkyl or benzyl.

The (S) -enantiomers of 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5- fluoro-1H-indol-3-yl) propionic acid, 2- (biphenyl) -4-sulfonylamino) -3- (5-fluoro-1H-indole- 3-yl) propionic acid, 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-hydroxy-1H-indol-3yl) - propionic acid, 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (1H-indol-3-yl) propionic acid or 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-methoxy-1H-indole-3-propionic acid.

The term "alkyl" is understood to mean a hydrocarbon residue Carbon chain is straight or branched. The raw materials of the chemical  Implementations are known or can be done using methods known from the literature easy to manufacture.

The invention further relates to a process for the preparation of the S-enantiomer of the compound of the formula I and / or a physiologically tolerable salt of the S-enantiomer of the compound of the formula I, which is characterized in that

• a) the S enantiomer of a tryptophan derivative of the formula II,
  wherein R ^{2 is} as defined in formula I, with a sulfonic acid derivative of formula III
  in which R ^{1 is} as defined in formula I and Y represents a halogen atom, imidazoyl or -OR ³ , in which R ^{3 represents} hydrogen atom, (C ₁ -C ₃ ) -alkyl, phenyl or benzyl, optionally substituted,
  in the presence of a base or, if appropriate, a dehydrating agent, to give a compound of the formula I, and if appropriate
• b) the compound of formula I obtained in the corresponding physiological tolerated salt.

The S-form of tryptophan, which is substituted by R ^2, serves as starting compounds of the formula II.

The starting products used to prepare the sulfonic acid derivatives of the formula III are preferably sulfonic acids or their salts of the formula IV, for example

The arylsulfonic acids of the formula IV are preferably used the one in Houben / Weyl "Methods of Organic Chemistry" Volume 9, pp. 450-546 described sulfonation with concentrated sulfuric acid optionally in the presence of a catalyst, sulfur trioxide and its Addition compounds or halogen sulfonic acids, such as chlorosulfonic acid.

As sulfonic acid derivatives according to formula III in particular find the Sulfonic acid chlorides use. The corresponding are used in their manufacture Sulphonic acids, also in the form of their salts such as sodium, ammonium or Pyridine salts in a known manner with phosphorus oxychloride or an inert Solvents such as methylene chloride, cyclohexane or chloroform in general Reaction temperatures from 20 ° C to the boiling point of the used Reaction medium implemented.

The reaction of the sulfonic acid derivatives of the formula III with a tryptophan Formula II according to process variant a), advantageously runs according to the type of bulkhead Baumann reaction. Alkali metal hydroxides such as are particularly suitable for this purpose Sodium hydroxide, but also alkali metal acetates, bicarbonates, carbonates and Amines. The reaction takes place in water or in a water miscible or immiscible solvents such as tetrahydrofuran (THF), acetone, or dioxane Acetonitrile instead, the reaction temperature generally from -10 ° C to 50 ° C. is held. In the event that the reaction is carried out in an anhydrous medium is found mainly tetrahydrofuran or methylene chloride, acetonitrile or dioxane in the presence of a base such as triethylamine, N-methylmorpholine, N-ethyl or  Use of diisopropyl etheramine, possibly in the presence of N, N- Dimethylaminopyridine as a catalyst.

The production of physiologically compatible salts from those capable of salt formation Compounds of the formula I are carried out in a manner known per se. The carboxylic acids form with basic reagents such as hydroxides, carbonates, Bicarbonates, alcoholates and ammonia or organic bases for example trimethyl or triethylamine, ethanolamine or triethanolamine or also basic amino acids, such as lysine, ornithine or arginine, stable alkali Alkaline earth metal or optionally substituted ammonium salts. If the Compounds of formula I having basic groups can be with strong Acids also produce stable acid addition salts. For this come both inorganic and organic acids such as hydrogen chloride, hydrogen bromide, Sulfur, phosphorus, methanesulfone, benzenesulfone, p-ToOluolsulfon-, 4- Bromobenzene sulfone, cyclohexylamido sulfone, trifluoromethyl sulfone, vinegar, oxal, Tartaric, succinic or trifluoroacetic acid in question.

The invention also relates to medicinal products characterized by an effective one Content of at least one S-enantiomer of the compound of formula I and / or a physiologically acceptable salt of the S enantiomer of the compound of Formula I together with a pharmaceutically suitable and physiological compatible carrier, additive and / or other active substances and auxiliaries.

Due to the pharmacological properties, the Compounds according to the invention for the prophylaxis and therapy of all such Diseases, on the course of which an increased activity of matrix-degrading Metalloproteinases is involved. This includes degenerative joint diseases such as osteoarthritis, spondylosis, cartilage loss after joint trauma or prolonged immobilization of the joints after meniscus or patella injuries or Torn ligaments. It also includes connective tissue diseases such as Collagenosis, periodontal diseases, wound healing disorders and chronic Diseases of the musculoskeletal system such as inflammatory, immunological or Metabolic acute and chronic arthrides, arthropathies, myalgias and  Bone Metabolism Disorders. The compounds of Formula I for the treatment of ulceration, atherosclerosis and stenosis. Farther the compounds of the formula I are suitable for the treatment of inflammation, Cancer, tumor metastasis, cachexia, anorexia and septic Shock.

The pharmaceuticals according to the invention are generally orally or parenterally administered. Rectal or transdermal application is also possible.

The invention also relates to a method for producing a medicament which characterized in that at least one compound of formula I with a pharmaceutically acceptable and physiologically acceptable carrier and optionally other suitable active ingredients, additives or auxiliaries in one brings suitable dosage form.

Suitable solid or galenical forms of preparation are, for example Granules, powders, coated tablets, tablets (micro) capsules, suppositories, syrups, juices, Suspensions, emulsions, tropics or injectable solutions as well as preparations with protracted release of active ingredient, in the manufacture of conventional auxiliaries such as Carriers, explosives, binders, coatings, swelling agents, lubricants or lubricants Flavors, sweeteners and solubilizers find use. As Commonly used auxiliaries are magnesium carbonate, titanium dioxide, lactose Mannitol and other sugars, talc, milk protein, gelatin, starch, cellulose and their derivatives, animal and vegetable oils such as cod liver oil, sunflower, peanut or sesame oil, polyethylene glycol and solvents such as sterile water and monohydric or polyhydric alcohols such as glycerin.

The pharmaceutical preparations are preferably in dosage units manufactured and administered, each unit as an active ingredient contains certain dose of the compound of formula I according to the invention. At solid dosage units such as tablets, capsules, coated tablets or suppositories, this dose can be up to about 1000 mg, but preferably about 5 to 300 mg and at  Injection solutions in ampoule form up to about 300 mg, but preferably about 10 up to 100 mg.

For the treatment of an adult patient weighing approximately 70 kg are - each after effectiveness of the compounds according to formula I, daily doses of about 5 mg up to 1000 mg of active ingredient, preferably about 20 to 300 mg. In certain circumstances however, higher or lower daily doses may also be appropriate. The Administration of the daily dose can be done both by single entry in the form of a single dosage unit or several smaller dosage units than also done by multiple doses divided at certain intervals.

¹ H-NMR spectra were recorded on a 400 MHz device from Bruker, usually with tetramethylsilane (TMS) as the internal standard and at room temperature (RT). The solvents used are given in each case. End products are usually determined by mass spectroscopic methods (FAB-, ESI-MS). Temperatures in degrees Celsius, RT means room temperature (22-26 ° C). Abbreviations used are either explained or correspond to the usual conventions.

example 1 (R, S) - (2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-fluoro-1H-indol-3-yl) - propionic acid

35 g (R, S) -5-fluortryptophan (157 mmol) were suspended in 100 ml of tetrahydrofuran (THF) at 0 ° C. A pH of 10 was set using an autotitrator with 2 molar sodium hydroxide solution. The solution of 47.5 g (165 mmol) of 4'-chlorobiphenyl-4-sulfonic acid chloride in 100 ml of THF was slowly added dropwise with automatic pH maintenance. After 3 h at 0 ° C., the mixture was stirred overnight at room temperature (RT). THF was evaporated on a rotary evaporator, the remaining suspension was covered with ethyl acetate and adjusted to pH 2-3 with 1 molar aqueous HCl at 0 ° C. The ethyl acetate phase was separated off, the aqueous phase was extracted twice more with ethyl acetate and the combined ethyl acetate phases were dried over sodium sulfate. After the desiccant had been removed and the solvent had been evaporated under reduced pressure, 59 g of a beige-colored residue remained. To remove non-polar secondary components, this was mixed with diethyl ether and stirred at RT overnight. The residue was 95.9% pure. To remove polar constituents further, it was taken up in methanol and precipitated by slowly adding water. Drying was carried out with an oil pump vacuum.
Yield: 49.5 g, 67% of theory
Purity (HPLC): <98%
¹ H NMR: (DMSO-d ₆ , 400 MHz, in ppm) 12.71 (s, 1 H); 10.88 (s, 1H);
8.30 (d, 1H); 7.715 (d. 2H); 7.58 (m, 7H); 7.17 (m. 2H);
7.05 (dd, 1H); 6.79 (m, 1H); 3.90 (m, 1H); 3.04 (m, 1H);
2.85 (m, 1 H)
MS (M⁺): 473.1

Example 2 (S) -2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-fluoro-1H-indol-3-yl) - propionic acid

The S isomer was used instead of the (R, S) -5-fluortryptophan. With the same reaction procedure as described in Example 1, a compound with the following properties was obtained:
Purity (HPLC): <99%
¹ H NMR: (DMSO-d ₆ , 400 MHz, in ppm) 12.71 (s, 1 H); 10.87 (s, 1H);
8.29 (d, 1H); 7.70 (d, 2H); 7.58 (m, 7H); 7.17 (m. 2H);
7.07 (dd, 1H); 6.79 (m, 1H); 3.90 (m, 1H); 3.04 (m, 1H);
2.85 (m, 1 H)
MS (M⁺): 473.2
Rotation value: α D ²⁰ = -46.1 ° (c = 1, methanol (MeOH))
Determination of the enantiomeric excess: <99% (conditions: Daicel Chiralpak AD, 4 × 250 mm, 254 nm, isocratic 25% (ethanol + 0.1% trifluoroacetic acid (TFA), 75% (hexane + 0.1% TFA), RT

Example 3 (R) -2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-fluoro-1H-indol-3-yl) - propionic acid

The R isomer was used instead of the (R, S) -5-fluortryptophan. With the same reaction procedure as described in Example 1, a compound with the following properties was obtained:
Purity (HPLC): <97%
¹ H NMR: (DMSO-d ₆ , 400 MHz, in ppm) 12.71 (s, 1 H); 10.88 (s, 1H); 8.30 (d, 1H); 7.71 (d, 2H), 7.58 (m, 7H); 7.17 (m. 2H); 7.05 (dd, 1H); 6.79 (m, 1H); 3.89 (m, 1H); 3.04 (m, 1H); 2.85 (m, 1H)
Rotation value: α D ²⁰ = + 46.1 ° (c = 1, MeOH)
Determination of the enantiomeric excess: <97% (conditions: Daicel Chiralpak AD, manufacturer Daicel Chemical Industries, Ltd .; sales Mallinckrodt Baker, Groß-Gerau, Germany; 4 × 250 mm, 254 nm, isocratic 25% (ethanol + 0.1% (TFA), 75% (hexane + 0.1% TFA), RT
The following compounds were prepared analogously to Examples 1 to 3.

Pharmacological examples

Representation and determination of the enzymatic activity of the catalytic domain of human stromelysin and neutrophil collagenase.

The two enzymes stromelysin (MMP-3) and neutrophil collagenase (MMP-8) were presented according to Ye et al. (Biochemistry; 31 (1992) pages 11231-11235).

To measure the enzyme activity or the enzyme inhibitor effect, 70 ul buffer solution and 10 ul enzyme solution with 10 ul of a 10% (v / v) aqueous dimethyl sulfoxide solution, which optionally contains the enzyme inhibitor, are incubated for 15 minutes. After adding 10 μl of a 10% (v / v) aqueous dimethyl sulfoxide solution which contains 1 mmol / l of the substrate, the enzyme reaction is monitored by fluorescence spectroscopy (328 nm (ex) / 393 nm / em)). The enzyme activity is shown as the increase in extinction / minute. The IC ₅₀ values listed in Table 2 are determined as those inhibition concentrations which in each case lead to 50% inhibition of the enzyme.

The buffer solution contains 0.05% Brij (Sigma, Deisenhofen, Germany) as well as 0.1 mol / l Tris / HCl, 0.1 mol / l NaCI, 0.01 mol / l CaCl ₂ and 0.1 mol / l piperazine -N, N'-bis [2-ethanesulfonic acid] (pH = 6.5).

The enzyme solution contains 5 µg / ml of one of the Ye et al. enzyme domains shown. The substrate solution contains 1 mmol / l of the fluorogenic substrate (7-methoxycoumarin-4-yl) acetyl-Pro-Leu-Gly-Leu-3- (2'-, 4'-dinitrophenyl) -L-2,3-diaminopropionyl- Ala-Arg-NH ₂ (Bachem, Heidelberg, Germany).

Table 2 shows the results:

Table 2

Determination of the maximum drug concentration (C _max ) in blood plasma and the area under the curve (AUCD 0-72 h) within 72 hours in dogs and rabbits.

In Examples 1, 4, 8 and 9, the racemates were administered for the test, being used to detect the active substance concentration of the corresponding enantiomers prior separation via a chiral column was required.

For Examples 2, 3, 6 and 7 was used to determine the drug concentrations the enantiomerically pure forms are used in the blood plasma.

A) dog

Two or five male dogs (beagle), body weight about 16 to 24 kg, the received no food overnight but had free access to water for one drug application each in the subsequent investigations used. The drug compounds were administered orally in the form of a single dose suspended in 1% carboxymethyl cellulose (CMC). Dose administered: 6 mg of the respective compound per kg of body weight of the animal.

Blood samples (2.0 ml each) were taken from the jugular vein with normal 2.0 ml tips removed and immediately transferred to sample vessels that have a sufficient amount contained an anticoagulant. Sampling was carried out as follows: Before drug application 5, 10, 15, 25, 45, 60, 75, 90 minutes after Drug application and 2, 4, 6, 8, 24, 30, 48, 54 and 72 hours after Active substance application.

Protein precipitation was performed on the blood samples in the presence of an internal standard prior to quantification. The amount of active ingredient was analyzed using a high pressure liquid chromatography system (HPLC) using a chiral column (Daicel Chiralpak AD). The detection was carried out with a UV detector. The pharmacokinetic parameters (C _max , AUCD) from the concentration-time profiles were calculated using the software program TOPFIT.

B) rabbits

Two or five male rabbits (chinchilla bastard), body weight about 2.0 up to 4.5 kg, which received no food overnight but free access to water had for each drug application in the following Investigations used. The active ingredient compounds were orally in the form of a  Single dose suspended in 1% aqueous CMC. Dose administered: 15 mg of the respective compound per kg of body weight of the animal.

The blood sampling was done under A). The samples were taken at the following times:
Before drug application, 30 minutes after drug application, 1, 2, 3, 4, 6, 24, 30, 48, 54 and 72 hours after drug application.

The blood samples were analyzed as described under A).

Table 3 shows the results:

Table 3

Claims (8)

1. (S) -enantiomer of the compound of formula I.
and / or a physiologically acceptable salt of the S-enantiomer of the compound of formula I, wherein
R ^{1 represents} hydrogen atom, fluorine, chlorine or bromine and
R ^{2 represents} hydrogen atom, fluorine, chlorine, bromine or -OR ³ ,
wherein R ^{3 is} hydrogen, (C ₁ -C ₆ ) alkyl or benzyl. 2. (S) -enantiomer of the compound of formula I according to claim 1 from the group 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-fluoro-1H-indol-3-yl) propionic acid, 2- (biphenyl) -4-sulfonylamino) -3- (5-fluoro-1H-indol-3-yl) propionic acid, 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-hydroxy-1H-indol-3yl) propionic acid, 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (1H-indol-3-yl) propionic acid and 2- (4'-Chlorobiphenyl-4-sulfonylamino) -3- (5-methoxy-1H-indole-3-propionic acid. 3. (S) -enantiomer of 2- (4'-chlorobiphenyl-4-sulfonylamino) -3- (5-fluoro-1H-indole-3- yl) propionic acid. 4. A process for the preparation of the (S) -enantiomer of the compound of formula I according to one or more of claims 1 to 3, characterized in that

• a) the S enantiomer of the tryptophan derivative of the formula II
  wherein R ^{2 is} as defined in formula I, with a sulfonic acid derivative of formula III
  in which R ^{1 is} as defined in formula I and Y represents a halogen atom, imidazoyl or -OR ³ , in which R ^{3 represents} hydrogen atom, (C ₁ -C ₆ ) -alkyl, phenyl or benzyl, optionally substituted,
  in the presence of a base or, if appropriate, a dehydrating agent, to give a compound of the formula I, and if appropriate
• b) the compound of formula I obtained is converted into the corresponding physiologically acceptable salt.

5. Medicinal product, characterized by an effective content of at least one Compound of formula I according to one or more of claims 1 to 3 together with a pharmaceutically suitable and physiological compatible carrier, additive and / or other active substances and auxiliaries. 6. Use of at least one compound of the formula I according to one or several of claims 1 to 3, for the manufacture of medicaments for Prophylaxis and therapy of diseases, on the course of which an intensified Activity of matrix-degrading metalloproteinases is involved.   7. Use according to claim 6, for the treatment of degenerative Joint diseases such as osteoarthritis, spondylosis, cartilage loss Articular trauma or prolonged immobilization after meniscus or Patella injuries or torn ligaments, diseases of the connective tissue we Collagenosis, periodontal diseases, wound healing disorders and chronic musculoskeletal disorders such as inflammatory, acute and chronic arthritis caused by immunology or metabolism, Arthropathy, myalgia and disorders of bone metabolism, the Ulceration, atherosclerosis and stenosis, but also for the treatment of Inflammation, cancer, tumor metastasis, cachexia, Anorexia and septic shock. 8. A process for the manufacture of a medicament, characterized in that at least one compound of the formula I according to one or more of the Claims 1 to 3 with a pharmaceutically suitable and physiological compatible carriers and, if appropriate, other suitable active ingredients, additional or excipients in a suitable dosage form.