DE102007032344A1 - Prodrug derivatives of 1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3,4 -c] pyridine-3-carboxamide - Google Patents

Abstract

The present application relates to prodrug derivatives of 1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-c] pyridine-3-carboxamide, process for their preparation, their use for the treatment and / or prophylaxis of diseases and their use for the preparation of medicaments for the treatment and / or prophylaxis of diseases, in particular thromboembolic diseases.

Description

The present application relates to prodrug derivatives of 1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [ 3,4-c] pyridine-3-carboxamide, Process for their preparation, their use for treatment and / or Prophylaxis of diseases and their use for the production of medicaments for the treatment and / or prophylaxis of diseases, in particular of thromboembolic diseases.

Prodrugs are derivatives of an active substance which undergo a single or multistage biotransformation of enzymatic and / or chemical nature in vivo before the actual active substance is released. A prodrug residue is usually used to improve the property profile of the underlying drug [ Ettmayer et al., J. Med. Chem. 47, 2393 (2004). ]. In order to achieve an optimal profile of action, the design of the prodrug residue as well as the desired release mechanism must be tailored very precisely to the individual drug, the indication, the site of action and the route of administration. A large number of drugs are administered as prodrugs which have improved bioavailability over the underlying drug, for example, by improving physicochemical profile, especially solubility, active or passive absorption properties or tissue-specific distribution. From the extensive literature on prodrugs may be mentioned as an example: H. Bundgaard (Ed.), Design of Prodrugs: Bioreversible Derivatives for Various Functional Groups and Chemical Entities, Elsevier Science Publishers BV, 1985 ,

1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3,4-c] pyridine -3-carboxamide [Apixaban, compound (A)] is an orally active, direct inhibitor of serine protease factor Xa, which exerts an essential function in the regulation of blood coagulation. The compound is currently undergoing in-depth clinical trials as a potential new drug for the prevention and treatment of thromboembolic disease [ WO 03/026652 . WO 03/049681 ].

connection (A), however, has only limited solubility in water and physiological media, such as an intravenous Application of the drug is difficult. Object of the present invention was therefore the identification of derivatives or prodrugs of compound (A) which has an improved solubility in said Media possess and at the same time after application a controlled Release of the active substance (A) in the body of the patient allow.

In WO 2005/028473 are described as prodrugs for carboxamides Acyloxymethylcarbamat prodrugs of oxazolidinones, which serve to increase the oral bioavailability. In WO 01/00622 discloses acyl prodrugs of carbamate inhibitors of inosine 5'-monophosphate dehydrogenase. Another type of amide prodrugs that release the underlying drug through a multi-step activation mechanism is for oxazolidinones in WO 03/006440 described.

in welcher
R¹ für Wasserstoff oder C₁-C₄-Alkyl steht,
wobei Alkyl substituiert sein kann mit einem Substituenten, wobei der Substituent ausgewählt wird aus der Gruppe bestehend aus Hydroxy und C₁-C₄-Alkoxy,
R² für Wasserstoff oder C₁-C₄-Alkyl steht
und
L für eine C₁-C₄-Alkandiyl-Gruppe steht, in welcher eine CH₂-Gruppe gegen ein O-Atom ausgetauscht sein kann,
oder
L für eine Gruppe der Formel

steht, worin
* die Anknüpfstelle an das N-Atom ist,
# die Anknüpfstelle an das C-Atom ist,
R³ für die Seitengruppe einer natürlichen α-Aminosäure oder ihrer Homologe oder Isomere steht,
oder
R¹ und R³ sind über eine (CH₂)₃- oder (CH₂)₄-Gruppe verknüpft und bilden zusammen mit dem Stickstoff- bzw. Kohlenstoffatom, an das sie gebunden sind einen 5- bzw. 6-Ring,
R⁴ für Wasserstoff oder Methyl steht,
R⁵ für C₁-C₄-Alkyl steht
und
R⁶ für Wasserstoff oder C₁-C₄-Alkyl steht,
sowie ihre Salze, Solvate und Solvate der Salze.The present invention relates to compounds of the general formula

in which
R ^{1 is} hydrogen or C ₁ -C ₄ -alkyl,
wherein alkyl may be substituted with a substituent, wherein the substituent is selected from the group consisting of hydroxy and C ₁ -C ₄ alkoxy,
R ^{2 is} hydrogen or C ₁ -C ₄ alkyl
and
L is a C ₁ -C ₄ alkanediyl group in which a CH ₂ group can be exchanged for an O atom,
or
L for a group of the formula

stands in which
* is the point of attachment to the N atom,
# is the link to the C atom,
R ^{3 is} the side group of a natural α-amino acid or its homologs or isomers,
or
R ¹ and R ³ are linked via a (CH ₂ ) ₃ or (CH ₂ ) ₄ group and together with the nitrogen or carbon atom to which they are attached form a 5- or 6-membered ring,
R ^{4 is} hydrogen or methyl,
R ^{5 is} C ₁ -C ₄ alkyl
and
R ^{6 is} hydrogen or C ₁ -C ₄ -alkyl,
and their salts, solvates and solvates of the salts.

invention Compounds are the compounds of the formula (I) and their salts, Solvates and solvates of the salts, the compounds encompassed by formula (I) the following formulas and their salts, solvates and solvates the salts as well as those of formula (I), hereinafter referred to as exemplary embodiments mentioned compounds and their salts, solvates and solvates of Salts, as far as those of formula (I), hereinafter not already mentioned salts, solvates and solvates salts.

The Compounds according to the invention can depending on their structure in stereoisomeric forms (Enantiomers, diastereomers) exist. The invention therefore comprises the enantiomers or diastereomers and their respective mixtures. From such mixtures of enantiomers and / or diastereomers can the stereoisomerically uniform constituents in known Isolate way.

Provided the compounds of the invention in tautomers Forms may include the present invention all tautomeric forms.

Salts used in the context of the present invention are physiologically acceptable salts of the compounds according to the invention. Also included are salts which are not suitable for pharmaceutical applications themselves, but for example for the isolation or purification of the inventive Connections can be used.

physiological acceptable salts of the compounds of the invention include acid addition salts of mineral acids, Carboxylic acids and sulfonic acids, e.g. B. salts of Hydrochloric acid, hydrobromic acid, sulfuric acid, Phosphoric acid, methanesulfonic acid, ethanesulfonic acid, Toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, Acetic acid, trifluoroacetic acid, propionic acid, Lactic acid, tartaric acid, malic acid, Citric acid, fumaric acid, maleic acid and benzoic acid.

When Solvates are in the context of the invention, such forms of the invention Compounds referred to in solid or liquid Condition through coordination with solvent molecules to form a complex. Hydrates are a special form of solvates, where coordination takes place with water. As solvates are in the context of the present invention, hydrates are preferred.

Unless otherwise specified, in the context of the present invention, the substituents have the following meaning:
Alkyl per se and "Alk" and "alkyl" in alkoxy is a linear or branched alkyl radical having usually 1 to 4, preferably 1 or 2 carbon atoms, by way of example and preferably methyl, ethyl, n-propyl, isopropyl and tert-butyl. butyl.

alkoxy is exemplary and preferably methoxy, ethoxy, n-propoxy, isopropoxy and tert-butoxy.

The side group of an α-amino acid in the meaning of R ³ includes both the side groups of the naturally occurring α-amino acids and the side groups of homologues and isomers of these α-amino acids. The α-amino acid can be present both in the L and in the D configuration or else as a mixture of the L and D form. As side groups are exemplified: hydrogen (glycine), methyl (alanine), propan-2-yl (valine), propan-1-yl (norvaline), 2-methylpropan-1-yl (leucine), 1-methylpropane-1 -yl (isoleucine), butan-1-yl (norleucine), phenyl (2-phenylglycine), benzyl (phenylalanine), p-hydroxybenzyl (tyrosine), indol-3-ylmethyl (tryptophan), imidazol-4-ylmethyl (histidine ), Hydroxymethyl (serine), 2-hydroxyethyl (homoserine), 1-hydroxyethyl (threonine), mercaptomethyl (cysteine), methylthiomethyl (S-methylcysteine), 2-mercaptoethyl (homocysteine), 2-methylthioethyl (methionine), carbamoylmethyl (asparagine ), 2-carbamoylethyl (glutamine), carboxymethyl (aspartic acid), 2-carboxyethyl (glutamic acid), 4-aminobutan-1-yl (lysine), 4-amine-3-hydroxybutan-1-yl (hydroxylysine), 3-aminopropane 1-yl (ornithine), 3-guanidinopropan-1-yl (arginine), 3-ureidopropan-1-yl (citrulline). Preferred α-amino acid side groups in the meaning of R ² are hydrogen (glycine), methyl (alanine), propan-2-yl (valine), propan-1-yl (norvaline), imidazol-4-ylmethyl (histidine), Hydroxymethyl (serine), 1-hydroxyethyl (threonine), carbamoylmethyl (asparagine), 2-carbamoylethyl (glutamine), 4-aminobutan-1-yl (lysine), 3-aminopropan-1-yl (ornithine), 3-guanidinopropane 1-yl (arginine). The L configuration is preferred.

If Residues substituted in the compounds of the invention Unless otherwise specified, the radicals may be be mono- or polysubstituted. In the context of the present Invention is that for all residues that occur several times, whose meaning is independent of each other. A substitution with one or two identical or different substituents prefers. Most preferably, the substitution with a Substituents.

In the formulas of the group which may stand for L, the end point of the line next to each of which a * or # is not a carbon atom or a CH ₂ group but is part of the bond to the atom to which L is bound is.

steht, worin
* die Anknüpfstelle an das N-Atom ist,
# die Anknüpfstelle an das C-Atom ist,
R³ für Wasserstoff, Methyl, Propan-2-yl, Propan-1-yl, Imidazol-4-ylmethyl, Hydroxymethyl, 1-Hydroxyethyl, Carbamoylmethyl, 2-Carbamoylethyl, 4-Aminobutan-1-yl, 3-Aminopropan-1-yl oder 3-Guanidinopropan-1-yl steht,
oder
R¹ und R³ sind über eine (CH₂)₃- oder (CH₂)₄-Gruppe verknüpft und bilden zusammen mit dem Stickstoff- bzw. Kohlenstoffatom, an das sie gebunden sind einen 5- bzw. 6-Ring,
R⁴ für Wasserstoff oder Methyl steht,
R⁵ für Methyl steht
und
R⁶ für Wasserstoff oder Methyl steht,
sowie ihre Salze, Solvate und Solvate der Salze.Preference is given to compounds of the formula (I) in which
R ^{1 is} hydrogen or C ₁ -C ₄ -alkyl,
R ^{2 is} hydrogen
and
L is a C ₂ -C ₄ alkanediyl group,
or
L for a group of the formula

stands in which
* is the point of attachment to the N atom,
# is the link to the C atom,
R ^{3 is} hydrogen, methyl, propan-2-yl, propan-1-yl, imidazol-4-ylmethyl, hydroxymethyl, 1-hydroxyethyl, carbamoylmethyl, 2-carbamoylethyl, 4-aminobutan-1-yl, 3-aminopropane-1 is -yl or 3-guanidinopropan-1-yl,
or
R ¹ and R ³ are linked via a (CH ₂ ) ₃ or (CH ₂ ) ₄ group and together with the nitrogen or carbon atom to which they are attached form a 5- or 6-membered ring,
R ^{4 is} hydrogen or methyl,
R ^{5 is} methyl
and
R ^{6 is} hydrogen or methyl,
and their salts, solvates and solvates of the salts.

steht, worin
* die Anknüpfstelle an das N-Atom ist,
# die Anknüpfstelle an das C-Atom ist,
R³ für Wasserstoff, Methyl, Propan-2-yl, Propan-1-yl, Imidazol-4-ylmethyl, Hydroxymethyl, 1-Hydroxyethyl, Carbamoylmethyl, 2-Carbamoylethyl, 4-Aminobutan-1-yl, 3-Aminopropan-1-yl oder 3-Guanidinopropan-1-yl steht,
oder
R¹ und R³ sind über eine (CH₂)₃- oder (CH₂)₄-Gruppe verknüpft und bilden zusammen mit dem Stickstoff- bzw. Kohlenstoffatom, an das sie gebunden sind einen 5- bzw. 6-Ring,
R⁴ für Wasserstoff oder Methyl steht,
und
R⁶ für Wasserstoff oder Methyl steht,
sowie ihre Salze, Solvate und Solvate der Salze.Preference is also given to compounds of the formula (I) in which
R ^{1 is} hydrogen, methyl or n-butyl,
R ^{2 is} hydrogen
and
L is a CH ₂ CH ₂ group or a group of the formula

stands in which
* is the point of attachment to the N atom,
# is the link to the C atom,
R ^{3 is} hydrogen, methyl, propan-2-yl, propan-1-yl, imidazol-4-ylmethyl, hydroxymethyl, 1-hydroxyethyl, carbamoylmethyl, 2-carbamoylethyl, 4-aminobutan-1-yl, 3-aminopropane-1 is -yl or 3-guanidinopropan-1-yl,
or
R ¹ and R ³ are linked via a (CH ₂ ) ₃ or (CH ₂ ) ₄ group and together with the nitrogen or carbon atom to which they are attached form a 5- or 6-membered ring,
R ^{4 is} hydrogen or methyl,
and
R ^{6 is} hydrogen or methyl,
and their salts, solvates and solvates of the salts.

Preference is also given to compounds of the formula (I) in which R ^{1 is} hydrogen or methyl.

Preference is also given to compounds of the formula (I) in which R ^{2 is} hydrogen.

Preference is also given to compounds of the formula (I) in which R ^{3 is} hydrogen, methyl, propan-2-yl, propan-1-yl, imidazol-4-ylmethyl, hydroxymethyl, 1-hydroxyethyl, carbamoylmethyl, 2-carbamoylethyl, 4 -Aminobutan-1-yl, 3-aminopropan-1-yl or 3-guanidinopropan-1-yl.

Preference is also given to compounds of the formula (I) in which R ^{4 is} hydrogen.

• [A] die Verbindung
  
  zunächst in einem inerten Lösungsmittel in Gegenwart einer Base mit einer Verbindung der Formel
  
  in welcher R² die oben angegebene Bedeutung hat und Q für eine Abgangsgruppe wie beispielsweise Chlor, Brom oder Iod steht, in eine Verbindung der Formel
  
  in welcher Q und R² die oben angegebenen Bedeutungen haben, überführt, diese dann in einem inerten Lösungsmittel mit dem Caesiumsalz einer α-Aminocarbonsäure der Formel
  
  in welcher R¹, R³ und R⁴ jeweils die oben angegebenen Bedeutungen haben, PG für eine Amino-Schutzgruppe wie beispielsweise tert.-Butoxycarbonyl (Boc) oder Benzyloxycarbonyl (Z) steht zu einer Verbindung der Formel
  
  in welcher R¹, R², R³, R⁴ und PG jeweils die oben angegebenen Bedeutungen haben, umsetzt und nachfolgend die Schutzgruppe PG nach üblichen Methoden unter Erhalt einer Verbindung der Formel
  
  in welcher R¹, R², R³ und R⁴ jeweils die oben angegebenen Bedeutungen haben, entfernt oder
• [B] Verbindung (A) in einem inerten Lösungsmittel in Gegenwart einer Base mit einer Verbindung der Formel
  
  in welcher PG die oben angegebene Bedeutung hat, R^1A für C₁-C₄-Alkyl steht, wobei Alkyl substituiert sein kann mit einem Substituenten, wobei der Substituent ausgewählt wird aus der Gruppe bestehend aus Hydroxy und C₁-C₄-Alkoxy, und L¹ für eine C₁-C₄-Alkandiyl-Gruppe steht, in welcher eine CH₂-Gruppe gegen ein O-Atom ausgetauscht sein kann, zu einer Verbindung der Formel
  
  in welcher R^1A, L¹ und PG jeweils die oben angegebenen Bedeutungen haben, umsetzt und anschließend die Schutzgruppe PG nach üblichen Methoden unter Erhalt einer Verbindung der Formel
  
  in welcher R^1A und L¹ die oben angegebenen Bedeutungen haben, entfernt oder
• [C] Verbindung (A) in einem inerten Lösungsmittel in Gegenwart einer Base mit einer Verbindung der Formel
  
  in welcher L¹ für eine C₁-C₄-Alkandiyl-Gruppe steht, in welcher eine CH₂-Gruppe gegen ein O-Atom ausgetauscht sein kann, und PG¹ und PG² unabhängig voneinander für eine Amino-Schutzgruppe wie beispielsweise tert.-Butoxycarbonyl (Boc), Benzyloxycarbonyl (Z) oder p-Methoxybenzyl (PMB) stehen und gleich oder verschieden sein können, zu einer Verbindung der Formel
  
  in welcher L¹, PG¹ und PG² jeweils die oben angegebenen Bedeutungen haben, umsetzt und anschließend die Schutzgruppen PG¹ und PG² nach üblichen Methoden, gleichzeitig oder sequentiell, unter Erhalt einer Verbindung der Formel
  
  in welcher L¹ die oben angegebene Bedeutung hat, entfernt und die jeweils resultierenden Verbindungen der Formel (I-A), (I-B) bzw. (I-C) gegebenenfalls mit den entsprechenden (i) Lösungsmitteln und/oder (ii) Säuren in ihre Solvate, Salze und/oder Solvate der Salze überführt.

Another object of the invention is a process for the preparation of the compounds of formula (I), characterized in that either

• [A] the connection
  
  first in an inert solvent in the presence of a base with a compound of the formula
  
  in which R ^{2 has} the abovementioned meaning and Q is a leaving group such as chlorine, bromine or iodine, into a compound of formula
  
  in which Q and R ^{2 have} the meanings given above, these are then converted in an inert solvent with the cesium salt of an α-aminocarboxylic acid of the formula
  
  in which R ¹ , R ³ and R ^{4 are} each as defined above, PG is an amino-protecting group such as tert-butoxycarbonyl (Boc) or benzyloxycarbonyl (Z) is a compound of formula
  
  in which R ¹ , R ² , R ³ , R ⁴ and PG each have the meanings given above, and subsequently the protective group PG by customary methods to obtain a compound of the formula
  
  in which R ¹ , R ² , R ³ and R ⁴ each have the meanings given above, removed or
• [B] Compound (A) in an inert solvent in the presence of a base with a compound of the formula
  
  in which PG has the abovementioned meaning, R ^{1A is} C ₁ -C ₄ -alkyl, where alkyl may be substituted by a substituent, where the substituent is selected from the group consisting of hydroxy and C ₁ -C ₄ -alkoxy, and L ^{1 is} a C ₁ -C ₄ alkanediyl group in which a CH ₂ group may be replaced by an O atom to a compound of the formula
  
  in which R ^1A , L ¹ and PG each have the meanings given above, and then the protective group PG by customary methods to obtain a compound of the formula
  
  in which R ^1A and L ^{1 have} the meanings given above, removed or
• [C] Compound (A) in an inert solvent in the presence of a base with a compound of the formula
  
  in which L ^{1 is} a C ₁ -C ₄ alkanediyl group in which a CH ₂ group may be replaced by an O atom, and PG ¹ and PG ² independently of one another for an amino-protecting group such as tert. Butoxycarbonyl (Boc), benzyloxycarbonyl (Z) or p-methoxybenzyl (PMB) and may be the same or different, to a compound of formula
  
  in which L ¹ , PG ¹ and PG ² each have the meanings given above, and then the protecting groups PG ¹ and PG ² by conventional methods, simultaneously or sequentially, to obtain a compound of formula
  
  in which L ^{1 has} the abovementioned meaning, and the respectively resulting compounds of the formula (IA), (IB) or (IC) optionally with the corresponding (i) solvents and / or (ii) acids in their solvates, salts and / or solvates of the salts.

The Compounds of formulas (I-A), (I-B) and (I-C) can in the preparation according to the methods described above also directly arise in the form of their salts. These salts may optionally by treatment with a base in an inert solvent chromatographic methods or by means of ion exchange resins be transferred to the respective free bases.

Functional groups optionally present in the radicals R ¹ , R ^1A and / or R ^{3 may also} be present in temporarily protected form in the case of the reaction sequences described above, if appropriate or necessary. The introduction and removal of such protective groups as well as the protective groups PG, PG ¹ and PG ² is carried out here by customary methods known from peptide chemistry [see, for example, US Pat. B. TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, Wiley, New York, 1999 ; M. Bodanszky and A. Bodanszky, The Practice of Peptide Synthesis, Springer-Verlag, Berlin, 1984 ].

Such protective groups optionally present in R ¹ , R ^1A and / or R ³ can be removed simultaneously with the removal of PG or in a separate reaction step before or after the removal of PG.

As the amino-protecting group PG, PG ¹ or PG ² , tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Z) or p-methoxybenzyl (PMB) is preferably used in the above processes. The cleavage of these protecting groups is carried out by conventional methods, preferably by reaction with a strong acid such as hydrogen chloride, hydrogen bromide or trifluoroacetic acid in an inert solvent such as dioxane, dichloromethane or acetic acid; if appropriate, the cleavage can also be carried out without an additional inert solvent.

As inert solvents, in the process steps (A) + (II) → (III), (A) + (VI) → (VII) and (A) + (VIII) → (IX) preferably uses tetrahydrofuran, N, N-dimethylformamide or dimethylsulfoxide; particularly preferred is N, N-dimethylformamide. In particular, sodium hydride is suitable as the base in these reactions. The reactions mentioned are generally carried out in a temperature range from 0 ° C to + 40 ° C at atmospheric pressure.

Of the Process step (III) + (IV) → (V) is preferably carried out in N, N-dimethylformamide as solvent. In general the reaction is in a temperature range from 0 ° C to + 50 ° C, preferably at + 20 ° C to + 50 ° C, carried out at atmospheric pressure. The implementation can also be beneficial be carried out under ultrasound treatment.

The compounds of the formulas (II), (IV), (VI) and (VIII) are commercially available, known from the literature or can be prepared by literature methods. The preparation of the compounds (A) is in WO 03/026652 and WO 03/049681 described.

Syntheseschema 2

The preparation of the compounds of the invention can be illustrated by the following synthesis schemes: Synthetic Scheme 1

Synthesis Scheme 2

The Compounds of the invention and their salts provide useful prodrugs of the active ingredient compound (A) On the one hand, they show good stability, for example at pH 4 and, on the other hand, show efficient conversion to the active ingredient compound (A) in vivo. In addition, own the compounds of the invention a good solubility in water and other physiologically compatible media, what they are for therapeutic use in particular suitable for intravenous administration.

Another The present invention is the use of the invention Compounds for the treatment and / or prophylaxis of diseases, preferably of thromboembolic disorders and / or thromboembolic Complications.

To the "thromboembolic diseases" within the meaning of the present Invention include in particular diseases such as myocardial infarction with ST segment increase (STEMI) and without ST segment increase (non-STEMI), stable angina pectoris, unstable angina pectoris, reocclusions and restenosis following coronary interventions such as angioplasty or aortocoronary bypass, peripheral arterial occlusive diseases, pulmonary embolism, deep venous thrombosis and renal vein thrombosis, transitory ischemic attacks as well as thrombotic and thromboembolic Stroke.

The Substances are therefore also suitable for prevention and treatment cardiogenic thromboembolism, such as brain ischemia, Stroke and systemic thromboembolism and ischemia, in patients with acute, intermittent or persistent Cardiac arrhythmias, such as atrial fibrillation, and such who undergo cardioversion, and in patients with Heart valve diseases or with artificial heart valves. In addition, the invention Compounds for the treatment of disseminated intravascular coagulation (DIC) suitable.

thromboembolic Complications also occur in microangiopathic hemolytic Anemias, extracorporeal blood circuits, such as Hemodialysis, as well as heart valve prostheses.

Furthermore the compounds according to the invention also come for the prophylaxis and / or treatment of atherosclerotic Vascular diseases and inflammatory diseases such as rheumatic diseases of the musculoskeletal system, beyond that also for prophylaxis and / or Treatment of Alzheimer's disease. In addition, you can the compounds of the invention for inhibition tumor growth and metastasis, in microangiopathies, age-related macular degeneration, diabetic retinopathy, diabetic nephropathy and other microvascular diseases as well as for the prevention and treatment of thromboembolic Complications such as venous thromboembolism Tumor patients, especially those who are larger undergo surgical procedures or chemo- or radiotherapy, be used.

Another The present invention is the use of the invention Compounds for the treatment and / or prophylaxis of diseases, in particular the aforementioned diseases.

Another The present invention is the use of the invention Compounds for the manufacture of a medicament for treatment and / or prophylaxis of diseases, in particular those mentioned above Diseases.

Another The subject of the present invention is a method of treatment and / or prophylaxis of diseases, in particular those mentioned above Diseases using the invention Links.

• • Lipidsenker, insbesondere HMG-CoA-(3-Hydroxy-3-methylglutaryl-Coenzym A)-Reduktase-Inhibitoren;
• • Koronartherapeutika/Vasodilatatoren, insbesondere ACE-(Angiotensin-Converting-Enzyme)-Inhibitoren; AII-(Angiotensin II)-Rezeptor-Antagonisten; β-Adrenozeptor-Antagonisten; alpha-1-Adrenozeptor-Antagonisten; Diuretika; Calciumkanal-Blocker; Substanzen, die eine Erhöhung von cyclischem Guanosinmonophosphat (cGMP) bewirken, wie beispielsweise Stimulatoren der löslichen Guanylatcyclase;
• • Plasminogen-Aktivatoren (Thrombolytika/Fibrinolytika) und die Thrombolyse/Fibrinolyse steigernde Verbindungen wie Inhibitoren des Plasminogen-Aktivator-Inhibitors (PAI-Inhibitoren) oder Inhibitoren des Thrombin-aktivierten Fibrinolyse-Inhibitors (TAFI-Inhibitoren);
• • antikoagulatorisch wirksame Substanzen (Antikoagulantien);
• • plättchenaggregationshemmende Substanzen (Plättchenaggregationshemmer, Thrombozytenaggregationshemmer);
• • Fibrinogen-Rezeptor-Antagonisten (Glycoprotein-IIb/IIIa-Antagonisten);
• • sowie Antiarrhythmika.

Another object of the present invention are pharmaceutical compositions containing a compound of the invention and one or more other active ingredients, in particular for the treatment and / or prophylaxis of the aforementioned diseases. As suitable combination active ingredients may be mentioned by way of example and preferably:

• • lipid-lowering drugs, in particular HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) reductase inhibitors;
• Coronary / vasodilators, especially angiotensin converting enzyme (ACE) inhibitors; AII (angiotensin II) receptor antagonists; β-adrenoceptor antagonists; alpha-1-adrenoceptor antagonists; diuretics; Calcium channel blockers; Substances that cause an increase in cyclic guanosine monophosphate (cGMP), such as soluble guanylate cyclase stimulators;
• Plasminogen activators (thrombolytics / fibrinolytics) and thrombolysis / fibrinolysis-enhancing compounds such as inhibitors of plasminogen activator inhibitor (PAI inhibitors) or inhibitors of Thrombin-activated fibrinolysis inhibitors (TAFI inhibitors);
• • anticoagulant substances (anticoagulants);
• • platelet aggregation inhibiting substances (platelet aggregation inhibitors, antiplatelet agents);
• Fibrinogen receptor antagonists (glycoprotein IIb / IIIa antagonists);
• • as well as antiarrhythmic drugs.

Another The present invention relates to medicaments which are at least a compound of the invention, usually together with one or more inert, non-toxic, pharmaceutical contain suitable excipients, as well as their use to the previously mentioned purposes.

The Compounds according to the invention can act systemically and / or locally. For this purpose can they are applied in a suitable manner, such as. Oral, parenteral, pulmonary or nasal. For these routes of application can the compounds of the invention in suitable Administration forms are administered.

For the oral administration are functional according to the prior art, the compounds of the invention quickly and / or modified donating application forms containing the compounds of the invention in crystalline and / or amorphised and / or dissolved Contain form such. As tablets (uncoated or coated Tablets, for example, with enteric or delayed dissolving or insoluble coatings that release control the compound of the invention), rapidly disintegrating tablets or films / wafers in the oral cavity, Films / lyophilisates, capsules (for example hard or soft gelatin capsules), Dragees, granules, pellets, powders, emulsions, suspensions, aerosols or solutions.

The Parenteral administration can bypass a resorption step happen (eg intravenous, intraarterial, intracardiac, intraspinal or intralumbar) or with involvement of absorption (eg, intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). Suitable for parenteral administration themselves as application forms u. a. Injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilisates or sterile powders.

For the other routes of administration are suitable for. B. inhalant dosage forms such as powder inhalers or nebulizers, or nasally administrable pharmaceutical forms like drops, solutions or sprays.

Prefers is the parenteral administration, especially the intravenous Application.

The Compounds according to the invention can converted into the mentioned application forms become. This can be done in a conventional manner by mixing with inert, Non-toxic, pharmaceutically suitable excipients happen. These adjuvants include u. a. excipients (for example, microcrystalline cellulose, lactose, mannitol), Solvents (eg liquid polyethylene glycols), Emulsifiers and dispersing or wetting agents (for example sodium dodecylsulphate, Polyoxysorbitanoleate), binders (for example polyvinylpyrrolidone), synthetic and natural polymers (for example albumin), Stabilizers (eg antioxidants such as ascorbic acid), Dyes (eg inorganic pigments such as iron oxides) and flavor and / or scent remedies.

in the In general, it has proven to be beneficial in parenteral Application amounts of about 0.001 to 1 mg / kg, preferably about 0.01 to 0.5 mg / kg body weight to achieve effective To give results. For oral administration is the dosage about 0.01 to 100 mg / kg, preferably about 0.01 to 20 mg / kg and most preferably 0.1 to 10 mg / kg of body weight.

Nevertheless it may be necessary, if necessary, of the quantities mentioned to deviate, depending on body weight, Application route, individual behavior towards the Active substance, method of preparation and time or interval, too which the application takes place. So it can in some cases be sufficient, with less than the aforementioned minimum quantity while in other cases the said upper limit must be exceeded. In case of application larger quantities may be recommended, this one in several single doses to distribute over the day.

The The following embodiments illustrate the Invention. The invention is not limited to the examples.

The Percentages in the following tests and examples are provided not stated otherwise, weight percentages; Parts are parts by weight. Solvent ratios, dilution ratios and concentration data of liquid / liquid solutions each refer to the volume.

A. Examples

Abbreviations and acronyms:

Section.

absolutely

Boc

tert-butoxycarbonyl

DMF

N, N-dimethylformamide

DMSO

dimethyl sulfoxide

d. Th.

the theory (at yield)

H

Hour (s)

HPLC

High pressure, high performance liquid chromatography

LC-MS

Liquid chromatography-coupled Mass spectrometry

min

Minute (s)

MS

Mass spectrometry

NMR

nuclear magnetic resonance spectrometry

Pd / C

Palladium on activated carbon

quant.

quantitative (at yield)

RT

room temperature

R

_t retention time (by HPLC)

UV

Ultraviolet spectrometry

v / v

Volume-to-volume ratio (a solution)

Z

benzyloxycarbonyl

LC-MS and HHPLC methods:

• Method 1 (LC-MS): Device Type MS: Micromass ZQ; Device type HPLC: Waters Alliance 2795; Column: Phenomenex Synergi 2μ Hydro-RP Mercury 20mm x 4mm; eluent A: 1 l of water + 0.5 ml of 50% formic acid, eluent B: 1 liter of acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; Flow: 0.0 min 1 ml / min 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 2 (LC-MS): Instrument: Micromass Quattro LCZ with HPLC Agilent 1100 series; Column: Phenomenex Synergi 2μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 liter of water + 0.5 ml of 50% Formic acid, eluent B: 1 l acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; River: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 208-400 nm.
• Method 3 (LC-MS): Device Type MS: Micromass ZQ; device type HPLC: HP 1100 Series; UV DAD; Column: Phenomenex Synergi 2 μ Hydro-RP Mercury 20 mm × 4 mm; Eluent A: 1 l water + 0.5 ml 50% formic acid, eluent B: 1 l acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; River: 0.0 min 1 ml / min 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 4 (LC-MS): Device Type MS: Micromass ZQ; device type HPLC: HP 1100 Series; UV DAD; Column: Phenomenex Gemini 3 μ 30 mm × 3.00 mm; Eluent A: 1 liter of water + 0.5 ml 50% formic acid, eluent B: 1 l acetonitrile + 0.5 ml of 50% formic acid; Gradient: 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5% A → 4.5 min 5% A; River: 0.0 min 1 ml / min → 2.5 min / 3.0 min / 4.5 min 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 5 (preparative HPLC): Device type HPLC: Abimed / Gilson Pump 305/306; Manometric Module 806; UV Knauer variable Wavelength Monitor; Column: Gromsil C18, 10 nm, 250 mm × 30 mm; Eluent A: 1 l of water + 0.5 ml of 99% trifluoroacetic acid, Eluent B: 1 liter acetonitrile; Gradient: 0.0 min 2% B → 10 min 2% B → 50 min 90% B; Flow: 20 ml / min; Volume: 628 ml A and 372 ml B.
• Method 6a (preparative HPLC): column: VP 250/21 Nuldeodur 100-5 C18 ec, Macherey & Nagel No. 762002; Eluent A: water / 0.01% trifluoroacetic acid, Eluent B: acetonitrile / 0.01% trifluoroacetic acid; Gradient: 0 min 0% B → 20 min 20% B → 40 min 20% B → 60 min 30% B → 80 min 30% B → 90 min 100% B → 132 min 100% B; Flow: 5 ml / min; Temperature: RT; UV detection: 210 nm.
• Method 6b (preparative HPLC): Column: VP 250/21 Nucleodur 100-5 C18 ec, Macherey & Nagel No. 762002; Eluent A: 1 L water / 1 mL 99% trifluoroacetic acid, eluent B: 1 L acetonitrile / 1 mL 99% trifluoroacetic acid; Gradient: 0 min 30% B → 20 min 50% B → 40 min 80% B → 60 min 100% B; Flow: 5 ml / min; Temperature: RT; UV detection: 210 nm.
• Method 7 (analytical HPLC): Column: XTerra 3.9 x 150 WAT 186000478; Eluent A: 10 ml of 70% perchloric acid in 2.5 liters of water, eluent B: acetonitrile; Gradient: 0.0 min 20% B → 1 min 20% B → min 90% B → 9 min 90% B; Temperature: RT; Flow: 1 ml / min.
• Method 8 (LC-MS): Instrument: Micromass Quattro LCZ with HPLC Agilent 1100 series; Column: Phenomenex Onyx Monolithic C18, 100 mm × 3 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, Eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 2 min 65% A → 4.5 min 5% A → 6 min 5% A; Flow: 2 ml / min; Oven: 40 ° C; UV detection: 208-400 nm.
• Method 9 (LC-MS): Instrument: Micromass Platform LCZ with HPLC Agilent 1100 series; Column: Thermo Hypersil GOLD 3 μ, 20 mm × 4 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, Eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 100% A → 0.2 min 100% A → 2.9 min 30% A → 3.1 min 10% A → 5.5 min 10% A; Oven: 50 ° C; Flow: 0.8 ml / min; UV detection: 210 nm.
• Method 10 (LC-MS): Device Type MS: Micromass ZQ; device type HPLC: Waters Alliance 2795; Column: Phenomenex Synergi 2.5 μ MAX-RP 100A Mercury 20 mm × 4 mm; Eluent A: 1 liter of water + 0.5 ml 50% formic acid, eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 0.1 min 90% A → 3.0 min 5% A → 4.0 min 5% A → 4.01 min 90% A; Flow: 2 ml / min; Oven: 50 ° C; UV detection: 210 nm.
• Method 11 (LC-MS): Instrument: Micromass Quattro LCZ with HPLC Agilent 1100 series; Column: Phenomenex Synergi 2.5 μ MAX-RP 100A Mercury 20 mm × 4 mm; Eluent A: 1 liter of water + 0.5 ml 50% formic acid, eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 0.1 min 90% A → 3.0 min 5% A → 4.0 min 5% A → 4.1 min 90% A; Flow: 2 ml / min; Oven: 50 ° C; UV detection: 208-400 nm.
• Method 12 (LC-MS): Instrument: Micromass QuattroPremier with Waters UPLC Acquity; Column: Thermo Hypersil GOLD 1.9 μ 50 mm × 1 mm; Eluent A: 1 l of water + 0.5 ml of 50% formic acid, Eluent B: 1 liter acetonitrile + 0.5 ml 50% formic acid; Gradient: 0.0 min 90% A → 0.1 min 90% A → 1.5 min 10% A → 2.2 min 10% A; Oven: 50 ° C; River: 0.33 ml / mm; UV detection: 210 nm.

NMR spectrometry:

NMR measurements are performed at a proton frequency of 400.13 MHz. The samples were usually dissolved in DMSO-d ₆ ; Temperature: 302 K.

starting compounds

The starting material is 1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3,4- c] pyridine-3-carboxamide [compound (A)] used, whose preparation is described elsewhere [ WO 03026652 . WO 03049681 ].

Example 1A

N- (chloroacetyl) -1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3 , 4-c] pyridine-3-carboxamide

130 mg (0.283 mmol) of 1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3 , 4-c] pyridine-3-carboxamide [Compound (A)] is dissolved under argon in 20 ml of absolute DMF. 20.4 mg (0.85 mmol) of sodium hydride are added and the mixture is stirred at RT for 20 min. Then 639 mg (5.66 mmol) of chloroacetyl chloride are added, keeping the reaction temperature at RT. After stirring at RT for 5 h, 25 ml of water are added with cooling. Then the solvent is removed in vacuo and the temperature should not rise above 25 ° C. The residue is taken up in 150 ml of dichloromethane and extracted by shaking three times with 100 ml of water. The organic phase is dried over sodium sulfate, filtered and concentrated. The residue is purified by flash chromatography on silica gel with toluene / ethanol 10: 1 as eluent. The appropriate fractions are combined and the solvent removed. The residue is lyophilized from dioxane / water 1: 1. 41 mg (27% of theory) of product are obtained.
HPLC (Method 7): R _t = 5.1 min;
LC-MS (Method 2): R _t = 2.22 min; m / z = 536 (M + H) ⁺

Example 2A

Benzyl- (4-chloro-4-oxobutyl) methyl-carbamate

First, 4 - [[(benzyloxy) carbonyl] (methyl) amino] butyric acid according to the literature [ Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) ] from commercially available 4 - {[(benzyloxy) carbonyl] amino} butyric acid. Alternatively, the preparation can also be carried out via the introduction of the benzyloxycarbonyl protective group into ω-N-methylaminoalkylcarboxylic acids obtained by P. Quitt et al. [Helv. Chim. Acta 46, 327 (1963)] are available, take place.

1.74 g (6.92 mmol) 4 - [[(benzyloxy) carbonyl] (methyl) amino] butyric acid are dissolved in 35 ml of dichloromethane and washed with 3.5 ml (48 mmol) of thionyl chloride. The mixture is heated for 1 h under reflux. It is then concentrated in vacuo, the residue added again with dichloromethane and again concentrated. Back remains a viscous oil which is dried under high vacuum. 1.8 g (96% of theory) of the target compound are obtained further reacted without further purification and characterization becomes.

Example 3A

Benzyl (5-chloro-5-oxopentyl) methylcarbamate

First becomes 5 - [[(benzyloxy) carbonyl] (methyl) amino] valeric acid as described for Example 2A according to known methods produced.

1.97 g (7.43 mmol) 5 - [[(benzyloxy) carbonyl] (methyl) amino] valeric acid are dissolved in 30 ml of dichloromethane and treated with 4.9 ml (67.3 mmol) of thionyl chloride. The mixture is heated for 1 h under reflux. It is then concentrated in vacuo, the residue added again with dichloromethane and again concentrated. Back remains a viscous oil which is dried under high vacuum. 2 g (95% of theory) of the target compound are obtained further reacted without further purification and characterization becomes.

Example 4A

Benzyl (3-chloro-3-oxopropyl) methyl-carbamate

First, 3 - [[(benzyloxy) carbonyl] (methyl) amino] propionic acid according to the literature [ Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) ] from commercially available 3 - {[(benzyloxy) carbonyl] amino} propionic acid. Alternatively, the preparation can also be carried out via the introduction of the benzyloxycarbonyl protective group into ω-N-methylaminoalkylcarboxylic acids obtained by P. Quitt et al. [Helv. Chim. Acta 46, 327 (1963)] are available, take place.

850 mg (3.58 mmol) of 3 - [[(benzyloxy) carbonyl] (methyl) amino] propionic acid are dissolved in 15 ml of dichloromethane and with 1.5 ml of oxalyl chloride added. The mixture is heated under reflux for 3 h. It is then concentrated in vacuo, the residue again with Dichloromethane added and concentrated again. Back stays a viscous oil which is dried in a high vacuum. you receives 915 mg (quant.) of the target compound, which without further purification and characterization is further implemented.

Example 5A

Benzyl- (6-chloro-6-oxohexyl) methyl-carbamate

First, 6 - [[(benzyloxy) carbonyl] (methyl) amino] caproic acid according to the literature [ Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) ] from commercially available 6 - {[(benzyloxy) carbonyl] amino} caproic acid. Alternatively, the preparation can also be carried out via the introduction of the benzyloxycarbonyl protective group into ω-N-methylaminoalkylcarboxylic acids obtained by P. Quitt et al. [Hely. Chim. Acta 46, 327 (1963) ] are available.

3850 mg (13.8 mmol) of 6 - [[(benzyloxy) carbonyl] (methyl) amino] caproic acid are dissolved in 60 ml of dichloromethane than dissolved and treated with 4 ml of oxalyl chloride. The mixture is heated under reflux for 3 h. It is then concentrated under reduced pressure, the residue is again combined with dichloromethane and concentrated again. What remains is a viscous oil, which is dried in a high vacuum. This gives 4.1 g (quant.) Of the target compound, which is further reacted without further purification and characterization.

embodiments

General Procedure 1 for the preparation of cesium salts of carboxylic acids or suitably protected amino acid derivatives:

1 mmol of the corresponding carboxylic acid is in a mixture from 10 ml of dioxane and 10 ml of water and dissolved with 0.5 mmol Cesium carbonate added. It is then lyophilized.

example 1

2 - [({1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro-1H-pyrazolo [3, 4-c] pyridin-3-yl} carbonyl) amino] -2-oxoethylglycinate trifluoroacetate

Stage a):

62 mg (116 μmol) of Example 1A are dissolved with 50 mg (162 μmol) of the cesium salt of Boc-glycine (prepared from Boc-glycine according to General Procedure 1) in 10 ml of DMF. Stir at RT overnight and then simmer. The remaining residue is purified by preparative HPLC (Method 6a). The appropriate fractions are combined, concentrated and the residue dried under high vacuum. The protected intermediate is isolated together with a by-product and initially used without further purification in the next stage.
HPLC (Method 7): R _t = 5.1 min;
LC-MS (Method 4): R _t = 2.54 min; m / z = 673 (MH) ^- .

Stage b):

21.2 mg (31 .mu.mol) of the protected intermediate obtained above are taken up in 5 ml of dichloromethane and mixed with 2 ml of anhydrous trifluoroacetic acid. After 30 min treatment in an ultrasonic bath at RT, the mixture is concentrated in vacuo. The remaining residue is purified by preparative HPLC (Method 6a). The appropriate fractions are combined, concentrated and the remaining residue is then lyophilized from dioxane. This gives 8.3 mg (11% of theory over 2 stages) of product.
HPLC (Method 7): R _t = 4.2 min;
LC-MS (method 1): R _t = 1.27 min; m / z = 575 (M + H) ⁺

Example 2

1- (4-Methoxyphenyl) -N- [4- (methylamino) butanoyl] -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro- 1H-pyrazolo [3,4-c] pyridine-3-carboxamide hydrochloride

Stage a):

100 mg (0.218 mmol) of compound (A) are dissolved in 10 ml of DMF, admixed with 16 mg (0.653 mmol) of sodium hydride, and the mixture is stirred for 30 min at RT. Then 587 mg (2.176 mmol) of Example 2A are added. The mixture is stirred for a further 30 min at RT and then the mixture is mixed with 1 ml of water. It is then concentrated and the residue is partitioned between 50 ml of ethyl acetate and 50 ml of water. The organic phase is separated off, shaken out three times with 10% strength sodium bicarbonate solution and then concentrated. The residue is purified by preparative HPLC (Method 6a). The appropriate fractions are combined, concentrated and dried under high vacuum. 62 mg (41% of theory) of the protected intermediate are obtained.
HPLC (Method 7): R _t = 5.4 min;
LC-MS (Method 12): R _t = 1.27 min; m / z = 693 (M + H) ⁺ .

Stage b):

55 mg (0.079 mmol) of the Z-protected intermediate obtained above are taken up in 10 ml of ethanol and 2 ml of glacial acetic acid and after addition of 100 mg of palladium on activated carbon (10% strength) hydrogenated at RT under atmospheric pressure for 2 h. The catalyst is filtered off and the filtrate is treated with 1 ml of 1N hydrochloric acid. It is then concentrated in vacuo and the residue is taken up in 30 ml of hydrochloric acid (pH 3). It is shaken twice with dichloromethane and once with ethyl acetate. The aqueous phase is then concentrated in vacuo to 2/3 of the volume and then lyophilized. 27 mg (57% of theory) of the title compound are obtained.
HPLC (method 7): R _t = 4.1 min;
LC-MS (Method 12): R _t = 0.8 min; m / z = 559 (M + H) ⁺ .

Example 3

1- (4-Methoxyphenyl) -N- [4- (methylamino) pentanoyl] -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro- 1H-pyrazolo [3,4-c] pyridine-3-carboxamide hydrochloride

Example 4

1- (4-Methoxyphenyl) -N- [4- (methylamino) propanoyl] -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro- 1H-pyrazolo [3,4-c] pyridine-3-carboxamide hydrochloride

Example 5

1- (4-Methoxyphenyl) -N- [4- (methylamino) hexanoyl] -7-oxo-6- [4- (2-oxopiperidin-1-yl) phenyl] -4,5,6,7-tetrahydro- 1H-pyrazolo [3,4-c] pyridine-3-carboxamide hydrochloride

B. Determination of solubility, Stability and release behavior

a) Determination of solubility:

The Test substance is dissolved in water pH 4 or in 5% dextrose solution pH 4 suspended. This suspension is left for 24 h at room temperature shaken. After ultra-centrifugation at 224000 g for 30 min, the supernatant is diluted with DMSO and analyzed by HPLC. Quantification is via a two-point calibration curve the test compound in DMSO.

HPLC method for acids:

Agilent 1100 with DAD (G1315A), quat. Pump (G1311A), autosampler CTC HTS PAL, degasser (G1322A) and column thermostat (G1316A); Pillar: Phenomenex Gemini C18, 5 μm, 50 mm × 2 mm; Temperature: 40 ° C; Eluent A: water / phosphoric acid pH 2, eluent B: acetonitrile; Flow rate: 0.7 ml / min; Gradient: 0-0.5 min 85% A, 15% B; Ramp 0.5-3 min 10% A, 90% B; 3-3.5 min 10% A, 90% B; Ramp 3.5-4 min 85% A, 15% B; 4-5 min 85% A, 15% B.

HPLC method for bases:

Agilent 1100 with DAD (G1315A), quat. Pump (G1311A), autosampler CTC HTS PAL, degasser (G1322A) and column thermostat (G1316A); Pillar: VDSoptilab Kromasil 100 C18, 3.5 μm, 60 mm × 2.1 mm; Temperature: 30 ° C; Eluent A: water + 5 ml perchloric acid / liter, Eluent B: acetonitrile; Flow rate: 0.75 ml / min; Gradient: 0-0.5 min 98% A, 2% B; Ramp 0.5-4.5 min 10% A, 90% B; 4.5-6 min 10% A, 90% B; Ramp 6.5-6.7 min 98% A, 2% B; 6.7-7.5 min 98% A, 2% B.

A Decomposition of example compounds in these solutions is not observed.

b) stability in buffer at different pH:

0.3 mg of the test substance are weighed in a 2 ml HPLC vial and added with 0.5 ml of acetonitrile. To release the substance the sample vial is for about 10 seconds placed in an ultrasonic bath. Then 0.5 ml added to the respective buffer solution and the sample treated again in an ultrasonic bath.

Used buffer solutions:

• pH 4.0: 1 liter of Millipore water is mixed with 1 N hydrochloric acid set to pH 4.0;
• pH 7.4: 90 g sodium chloride, 13.61 g potassium dihydrogen phosphate and 83.35 g of 1 N sodium hydroxide solution to 1 liter with Millipore water refilled.

about a period of 24 hours at 37 ° C will be hourly in each case 10 .mu.l of the sample solution by HPLC on their Content of unchanged test substance analyzed. quantified is about the area percent of the corresponding Peaks.

HPLC method:

Agilent 1100 with DAD (G1314A), binary pump (G1312A), autosampler (G1329A), column oven (G1316A), thermostat (G1330A); Pillar: Kromasil 100 C18, 125 mm × 4 mm, 5 μm; Column temperature: 30 ° C; Eluent A: water + 5 ml perchloric acid / liter, Eluent B: acetonitrile; Gradient: 0-3.0 mm 80% A, 20% B; 3.0-18.0 min 80% A, 20% B; 18.0-20.0 min 10% A, 90% B; 20.0-21.0 min 10% A, 90% B; 21.0-23.0 90% A, 10% B; 23.0-26.0 90% A, 10% B; Flow rate: 2.0 ml / min; UV detection: 278 nm.

Table 2 shows, for representative embodiments, the ratios of the peak areas (F) at the respective times in relation to the peak areas at the start time: TABLE 2 Example no. Buffer pH % Test substance after 4 h [F (t = 4 h) × 100 / F (t = 0 h)] % Test substance after 24 h [F (t = 24 h) × 100 / F (t = 0 h)] 1 4 99 95 1 7.4 0 0 2 4 100 96 2 7.4 0 0

c) in vitro stability in rat and human plasma:

1 mg of the test substance is weighed into a 2 ml HPLC vial and mixed with 1.0 ml of DMSO and 1.5 ml of water. To dissolve the substance, the sample vessel is placed in the ultrasonic bath for approx. 10 seconds. To 0.5 ml of this solution 0.5 ml 37 ° C warm rat or human plasma are given. The sample is shaken and removed for a first analysis about 10 .mu.l (time t ₀ ). In the period up to 2 hours after incubation 4-6 further aliquots are taken for quantification. The sample is kept at 37 ° C over the test time. Characterization and quantification are by HPLC.

HPLC method:

Agilent 1100 with DAD (G1314A), binary pump (G1312A), autosampler (G1329A), column oven (G1316A), thermostat (G1330A); Pillar: Kromasil 100 C18, 250 mm × 4 mm, 5 μm; Column temperature: 30 ° C; Eluent A: water + 5 ml perchloric acid / liter, Eluent B: acetonitrile; Gradient: 0-7.0 mm 71% A, 29% B; 7.0-18.0 min 71% A, 29% B; 18.0-20.0 min 10% A, 90% B; 20.0-21.0 min 10% A, 90% B; 21.0-22.5 min 98% A, 2% B; 22.5-25.0 min 98% A, 2% B; Flow rate: 2 ml / min; UV detection: 278 nm.

In Table 3, for representative embodiments, the respective times are indicated in which 50% of the maximum possible amount of active substance compound (A) have arisen (t _{50% A} ). For the evaluation, the ratio of the peak areas to the individual times with respect to the starting point is used. Table 3 Example no. t _{50% A} [min] (rat plasma) 1 2.0 2 5.0

d) i.v. pharmacokinetics in Wistar rats:

On the day before administration of the substance to the experimental animals (male Wistar rats, body weight 200-250 g) is implanted, a catheter for blood collection into the jugular vein under isoflurane ^® narcosis.

On the day of the test, a defined dose of the test substance is administered as a solution with a Hamilton ^® syringe into the tail vein (bolus administration, application time <10 s). Within 24 hours of substance administration, blood samples (8-12 times) are taken sequentially across the catheter. For plasma collection, the samples are centrifuged in heparinized tubes. At each point in time, a defined plasma volume for protein precipitation is mixed with acetonitrile. After centrifugation, test substance and optionally known cleavage products of the test substance are quantitatively determined in the supernatant with a suitable LC / MS-MS method.

The calculation of pharmacokinetic parameters of the test substance or of the active substance compound (A) released therefrom, such as AUC, C _max , T _1/2 (half-life) and CL (clearance), takes place from the measured plasma concentrations.

e) Determination of the antithrombotic effect in an arteriovenous shunt model in rats:

Fasting male rats (strain: HSD CPB: WU) are anesthetized by intraperitoneal administration of a Rompun / Ketavet solution (12 mg / kg / 50 mg / kg). The formation of thrombus in an arteriovenous shunt is based on that of PC Wong et al. method described [Thrombosis Research 83 (2), 117-126 (1996)]. ] triggered. For this purpose, the left jugular vein and the right carotid artery are dissected free. Into the artery is an 8 cm long polyethylene catheter (PE60, Becton-Dickinson), followed by a 6 cm long Tygon tube (R-3606, ID 3.2 mm, Kronlab), which is roughened to create a thrombogenic surface and closed a double loop nylon thread (60 x 0.26 mm, Berkley Trilene) included. In the jugular vein, a 2 cm long polyethylene catheter (PE60, Becton-Dickinson) is incorporated and connected via a 6 cm long polyethylene catheter (PE160, Becton-Dickinson) with the Tygonschlauch. The tubes are filled with saline before opening the shunt. The extracorporeal circuit is maintained for 15 minutes. Then the shunt is removed and the nylon thread with the thrombus weighed immediately. The net weight of the nylon thread was determined before the start of the test. The test substance (as a solution in physiological saline adjusted to pH 4 with 0.1N hydrochloric acid) is administered as a bolus injection prior to application of the extracorporeal circuit.

C. Embodiments for pharmaceutical compositions

The Compounds according to the invention can for example, converted into pharmaceutical preparations as follows become:

iv solution:

The Compound of the invention is in a concentration below the saturation solubility in a physiological compatible solvents (eg isotonic Saline, glucose solution 5% and / or PEG 400 solution 30%, each to a pH of 3-5 are set) solved. The solution is optionally sterile filtered and / or in sterile and pyrogen-free injection containers bottled.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• WO 03/026652 [0003, 0031]
• WO 03/049681 [0003, 0031]
• WO 2005/028473 [0005]
• WO 01/00622 [0005]
• WO 03/006440 [0005]
• WO 03026652 [0056]
• WO 03049681 [0056]

Cited non-patent literature

• Ettmayer et al., J. Med. Chem. 47, 2393 (2004) [0002]
• H. Bundgaard (Ed.), Design of Prodrugs: Bioreversible Derivatives for Various Functional Groups and Chemical Entities, Elsevier Science Publishers BV, 1985 [0002]
• TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, Wiley, New York, 1999. [0026]
• M. Bodanszky and A. Bodanszky, The Practice of Peptide Synthesis, Springer-Verlag, Berlin, 1984 [0026]
• - Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) [0058]
• - P. Quitt et al. [Helv. Chim. Acta 46, 327 (1963)] [0058]
• - Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) [0062]
• - P. Quitt et al. [Helv. Chim. Acta 46, 327 (1963)] [0062]
• Y. Aramaki et al., Chem. Pharm. Bull. 52, 258 (2004) [0064]
• - P. Quitt et al. [Hely. Chim. Acta 46, 327 (1963) [0064]
• PC Wong et al. method described [Thrombosis Research 83 (2), 117-126 (1996) [0085]

Claims (11)

Compound of the formula

in which R ^{1 is} hydrogen or C ₁ -C ₄ alkyl, wherein alkyl may be substituted with a substituent, wherein the substituent is selected from the group consisting of hydroxy and C ₁ -C ₄ alkoxy, R ^{2 is} hydrogen or C ₁ -C ₄ alkyl and L is a C ₁ -C ₄ alkanediyl group in which a CH ₂ group may be replaced by an O atom, or L is a group of the formula

where * is the point of attachment to the N atom, # is the point of attachment to the C atom, R ^{3 is} the side group of a natural α-amino acid or its homologs or isomers, or R ¹ and R ³ are above a ( CH ₂ ) ₃ - or (CH ₂ ) ₄ group and form together with the nitrogen or carbon atom to which they are attached a 5- or 6-membered ring, R ^{4 is} hydrogen or methyl, R ^{5 is} C ₁ -C ₄ -alkyl and R ^{6 is} hydrogen or C ₁ -C ₄ -alkyl, and their salts, solvates and solvates of the salts. A compound of the formula (I) according to claim 1, in which R ^{1 is} hydrogen or C ₁ -C ₄ -alkyl, R ^{2 is} hydrogen and L is a C ₂ -C ₄ -alkanediyl group, or L is a Group of formula

where * is the point of attachment to the N atom, # is the point of attachment to the C atom, R ^{3 is} hydrogen, methyl, propan-2-yl, propan-1-yl, imidazol-4-ylmethyl, hydroxymethyl, 1-hydroxyethyl, carbamoylmethyl, 2-carbamoylethyl, 4-aminobutan-1-yl, 3-aminopropan-1-yl or 3-guanidinopropan-1-yl, or R ¹ and R ³ are via a (CH ₂ ) ₃ - or (CH ₂ ) ₄ group and form together with the nitrogen or carbon atom to which they are attached a 5- or 6-membered ring, R ^{4 is} hydrogen or methyl, R ^{5 is} methyl and R ⁶ is hydrogen or methyl, and their salts, solvates and solvates of the salts. A compound of the formula (I) according to claim 1 or 2, in which R ^{1 is} hydrogen, methyl or n-butyl, R ^{2 is} hydrogen and L is a CH ₂ CH ₂ group or a group of the formula

where * is the point of attachment to the N atom, # is the point of attachment to the C atom, R ^{3 is} hydrogen, methyl, propan-2-yl, propan-1-yl, imidazol-4-ylmethyl, hydroxymethyl, 1-hydroxyethyl, carbamoylmethyl, 2-carbamoylethyl, 4-aminobutan-1-yl, 3-aminopropan-1-yl or 3-guanidinopropan-1-yl, or R ¹ and R ³ are via a (CH ₂ ) ₃ - or (CH ₂ ) ₄ group and form together with the nitrogen or carbon atom to which they are attached a 5- or 6-membered ring, R ^{4 is} hydrogen or methyl, and R ^{6 is} hydrogen or methyl , and their salts, solvates and solvates of salts Process for the preparation of a compound of the formula (I) or one of its salts, its solvates or the solvates of its salts according to Claim 1, characterized in that either [A] the compound

first in an inert solvent in the presence of a base with a compound of the formula

in which R ^{2 has} the meaning given in claim 1 and Q is a leaving group such as chlorine, bromine or iodine, in a compound of formula

in which Q and R ^{2 have} the meanings given above, these are then converted in an inert solvent with the cesium salt of an α-aminocarboxylic acid of the formula

in which R ¹ , R ³ and R ⁴ each have the meanings given in claim 1, PG is an amino-protecting group such as tert-butoxycarbonyl (Boc) or benzyloxycarbonyl (Z) is a compound of formula

in which R ¹ , R ² , R ³ , R ⁴ and PG each have the meanings given above, and subsequently the protective group PG by customary methods to obtain a compound of the formula

in which R ¹ , R ² , R ³ and R ^{4 are} each as defined in claim 1, or [B] compound (A) in an inert solvent in the presence of a base with a compound of formula

in which PG has the abovementioned meaning, R ^{1A is} C ₁ -C ₄ -alkyl, where alkyl may be substituted by a substituent, where the substituent is selected from the group consisting of hydroxy and C ₁ -C ₄ -alkoxy, and L ^{1 is} a C ₁ -C ₄ alkanediyl group in which a CH ₂ group may be replaced by an O atom to a compound of the formula

in which R ^1A , L ¹ and PG each have the meanings given above, and then the protective group PG by conventional methods to obtain a compound of formula

in which R ^1A and L ^{1 have} the meanings given above, or [C] Compound (A) in an inert solvent in the presence of a base with a compound of the formula

in which L ^{1 is} a C ₁ -C ₄ alkanediyl group in which a CH ₂ group may be replaced by an O atom, and PG ¹ and PG ² independently of one another for an amino-protecting group such as tert. Butoxycarbonyl (Boc), benzyloxycarbonyl (Z) or p-methoxybenzyl (PMB) and may be the same or different, to a compound of formula

in which L ¹ , PG ¹ and PG ² each have the meanings given above, and then the protecting groups PG ¹ and PG ² by conventional methods, simultaneously or sequentially, to obtain a compound of formula

in which L ^{1 has} the abovementioned meaning, and the respectively resulting compounds of the formula (IA), (IB) or (IC) optionally with the corresponding (i) solvents and / or (ii) acids in their solvates, salts and / or solvates of the salts A compound of the formula (I) as claimed in any one of the claims 1 to 3, for the treatment and / or prophylaxis of diseases. Use of a compound of formula (I) as in one of claims 1 to 3 defined for the production a drug for the treatment and / or prophylaxis of thromboembolic Diseases. Medicament containing a compound of the formula (I) as defined in any one of claims 1 to 3, optionally in combination with an inert, non-toxic, pharmaceutical suitable excipient. Medicament containing a compound of the formula (I) as defined in any one of claims 1 to 3, in combination with another active ingredient. Medicament according to claim 7 or 8 for the treatment and / or prophylaxis of thromboembolic disorders. Medicament according to one of claims 7 to 9 for intravenous use. Method for the treatment and / or prophylaxis of using thromboembolic diseases in humans and animals at least one compound of the formula (I) as in one of claims 1 to 3, or a drug as in any of the claims 7 to 10 defined.