HUT71796A - Condensed cyclic compounds with nitrogen heteroatoms and pharmaceutical compositions of fibrinogen receptor antagonistic activity containing them - Google Patents

Description

FIELD OF THE INVENTION The present invention relates to fibrinogen receptor antagonists which generally modulate cell adhesion and inhibit the binding of fibrinogen and other proteins to platelets and inhibit platelet aggregation specifically at the IIb / IIIa fibrinogen receptor sites. Fibrinogen is a glycoprotein that is present in the plasma and is involved in platelet aggregation and fibrin formation. Platelets are cellular nucleated fragments that are found in the blood of all mammals and are also involved in blood coagulation. Interaction of fibrinogen with the IIb / IIIa receptor is known to be essential for the normal formation of platelets.

When a blood vessel is damaged by injury or other factors, platelets adhere to the injured subendothelial surface. The adherent platelets will then release biologically active components and aggregate. Aggregation is initiated by binding of agonists such as thrombin, epinephrine or ADP to specific platelet membrane receptors. As a result of agonist stimulation, latent fibrinogen receptors on the platelet surface open and fibrinogen binding to the glycoprotein IIb / IIIa receptor complex occurs.

Many attempts have been made to use natural materials and synthetic peptides to determine the mechanism of adhesion and platelet aggregation. For example, Rouslahti and Pierschbacher, Science, 238, 491-497 (1987), disclose adhesive proteins such as fibronectin, vitronectin, osteopontin, collagens, thrombospondin, fibrinogen and von Willebrand factor present in extracellular matrix and . These proteins contain arginine-gly-3

cin aspartic acid (RGD) tripeptide as a recognition site for glycoptrotein IIb / IIIa. These tripeptides containing arginine, glycine and aspartic acid are recognized by at least one member of the family of structurally related receptors, the so-called. integrins, which are heterodimeric proteins that contain two membrane stretching subunits. The authors of that publication state that the nature of the tripeptide sequence may be critical to the specificity of recognition.

Cheresh, Proc. Nat'l Acad. Sci. 84, 6471-6475 (1987), discloses an Arg-Gly-Asp adhesion receptor expressed by human endothelial cells that is structurally similar to the platelet IIb / IIIa complex but is antigenically distinct and functionally distinct. This receptor is directly involved in the attachment of endothelial cells to fibrinogen, von Willebrand factor and vitronectin.

Pierschbacher and Rouslahti, J. of Bioi. Chem., 262, (36), 17294-17298 (1987), suggest that the Arg-Gly-Asp sequence alone would be sufficient for signal recognition and binding, and that the conformation of the tripeptide sequence would be nature. Various synthetic peptides have been prepared and the authors conclude that the stereochemical conformational change of the Arg-Gly-Asp enantiomeric substitutions or additions significantly influences the receptor-ligand interaction. The authors also demonstrated that cyclization of the decapeptide via a disulfide bridge between the non-terminal residues Pen and Cys results in the peptide being much less effective in inhibiting the binding to fibronectin.

-4A Proc. Nat'l Acad. Sci. U.S.A., 81, 5985-5988 (1984), describe the tetrapeptide-like variants of the fibronectin cell recognition site which retain their binding-promoting activity. U.S. Patent Nos. 4,589,881 and 4,614,517 also disclose peptides having a tetrapeptide recognition site. Many large polypeptide fragments have cell-associated activity in the cell binding domain of fibronectin. In this regard, reference is made to 4,517,686; U.S. Patent Nos. 4,661,111 and 4,578,079.

Ruggeri et al., Proc. Nat'l Acad. Sci. USA, 83, 5708-5712 (1986), describes a series of synthetic peptides consisting of 16 amino acid residues in length and containing RGD and valine bound to the aspartic acid residue of RGD, which inhibit fibrinogen binding. platelets. In this regard, reference is made to Koczewiak et al., Biochem., 23, 1767-1774 (1984), and Ginsberg et al., J. Biol. Chem., 260 (7), 3931-3936 (1985), and by Blood, 66 (4), 946-952 (1985), and Haverstick et al. Other inhibitors are known from European Publication Nos. 275,748 and 298,820.

Several low molecular weight polypeptide factors have been isolated from snake venom. Obviously, these factors show high activity for gpl1b / IIIa complexes. For example, Huang et al., J. Biol. Chem., 262, 16157-16163 (1987) and Biochemistry, 28, 661-666 (1989), describe the structure of a snake venom trigramin primer, a 72 amino acid polypeptide containing an RGD subunit. Another substance that can be distinguished from the snake venom is echistatin, which has a high affinity for the gplIb / IIIa complex. This polypeptide contains 49 amino acids and has an RGD subunit and various disulfide bridges [I. Gan et al., J. Bioi. Chem. 263: 19827-19832 (1988); Dennis et al., Proc. Nat'l Acad. Sci. U.S.A. 87: 2471-2475 (1989)]. However, these snake venom factors also have a high affinity for other members of the adhesive protein receptor family, including the vitronectin and fibronectin receptors, so that they are not selective for the gpIIb / IIIa complexes.

Although the Arg-Gly-Asp tripeptide sequence is known to be present in polypeptides capable of duplicating or inhibiting the cellular binding effect of fibronectin and vitronectin, the Arg-Gly-Asp tripeptide itself has little activity. At present, it is not entirely clear how other amino acids linked to this sequence affect the specificity of the binding. U.S. Patent No. 5,023,233 discloses small cyclic hexapeptides containing the Arg-Gly-Asp sequence and useful as inhibitors of platelet aggregation. U.S. Patent No. 5,037,808 discloses indole-type platelet aggregation inhibitors that are believed to act by inhibiting interactions between fibrinogen and / or extracellular matrix proteins and platelet gpl1b / IIIa receptor. U.S. Patent No. 5,037,808 also discloses compounds having a structure resembling the structure of a guanidino-peptide that contain an Asp residue and inhibit platelet aggregation. THE

International Application No. -6PCT / US90 / 02746 discloses the use of antibody-polypeptide conjugates wherein the polypeptides comprise the Arg-Gly-Asp (RGD) sequence.

PCT / US91 / 00564 discloses large cyclic peptides which contain RGD among proline residues and also inhibit platelet aggregation. PCT / US90 / 03788 discloses small ring platelet aggregation inhibitors which are synthetic cyclic pentapeptides having the Arg-Gly-Asp tripeptide sequence and a thioether bond in the ring. PCT / US90 / 05367 also discloses peptides and pseudopeptides, such as N-amidinopiperidine-3-carboxyglycyl-L-aspartyl-L-valine, which inhibit platelet aggregation and thrombus formation in mammalian blood. European Patent Application No. 91103462.7 discloses linear compounds which include piperazinyl or piperidinyl international derivatives. European Patent Application No. 91300179.8 (published July 17, 1991) discloses linear polypeptide-type fibrinogen receptor antagonists. European Patent Application 901001404.3 discloses compounds of the formula R ¹ -A- (W) _a -X- (CH ₂ ) b- (Y) c-B'Z-COOR, wherein R ¹ is a guanidino or amidino moiety. , and A and B specific monosubstituted aryl groups or heterocyclic groups.

Although many compounds or peptide analogs are believed to be inhibited by platelet aggregation by preventing the platelet from binding to fibrinogen, the present invention provides novel fibrinogen receptor antagonists that have significant binding activity and are therefore for very good reasons. A number of very serious diseases and disorders are known, in which hyperthrombotic complications occur, resulting in the formation of thrombi and embolus intravascularly. There is a need for new and effective fibrinogen receptor antagonists for the treatment of myocardial infarction, stroke, varicose veins and many other serious diseases.

It has now been found that novel compounds of formula I and II, such as compound of formula III, have such fibrinogen receptor antagonist activity.

Accordingly, the present invention relates to compounds of formula (I) and (II). In these formulas

Q is a group of formula (IV) or (V) or a group of formula (VI) or Q is a 4- to 9-membered mono- or bicyclic ring group selected from 1, 2 or 3 atoms selected from nitrogen, oxygen and sulfur atoms. containing a heteroatom and optionally substituted with R ⁸ as defined below;

AB is a condensed ring system containing adjacent carbon and nitrogen atoms in which

A is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; and

B is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen;

-8R ¹ is hydrogen, 1-4 alkyl, or -N ^(R8) 2, -N ^(R8) SO2R ^7, -NR ⁸ CO ₂ R ^7, -NR ⁸ C (O) R ^7, -NR ⁸ C (O) N (R ⁷ ) R ⁸ ,

-N (R ^e ) SO 2 N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁸ ) C (O) OR ⁷ , -C (O) N (R ⁷ ) ₂ or R ⁶ as defined below a cyclic ring;

R ² is hydrogen or C 1 -C 4 straight or branched alkyl, aryl (C 1 -C 4) alkyl or aryl;

R ⁴ is hydrogen or C 1-4 straight or branched chain or cyclic alkyl or C 1-4 alkenyl;

^R5 is CH, -CH _{(CH2) n} group, a bond or - when ^R5 is adjacent N ^(R4) group - of formula (VII) group;

R ⁶ is carboxyl, hydroxymethyl, tetrazolyl, acylsulfonamido, -C (O) N (R ⁷ ) ₂ , -CO ₂ R ⁹ , or (VIII) or R ⁶ R ¹ or a substituent of formula (IX) or (X);

y is oxygen or sulfur;

R ⁷ is hydrogen, or C 1-4 straight or branched, optionally substituted alkyl, straight or branched lower alkenyl, aryl (C 1-4) alkyl, substituted aryl, or 5- or 6-membered, Heteroaryl radicals containing 1, 2, or 3 nitrogen, sulfur or oxygen atoms as heteroatoms; wherein the substituted alkyl group is substituted by hydroxy or C 1 -C 4 alkoxy and the substituted aryl group has 1, 2 or 3 substituents selected from halogen, C 1 -C 4 alkoxy, hydroxy or C 1 -C 4 alkyl;

-9R ⁸ is hydrogen or 1-4 carbon atoms, straight or branched chain alkyl;

R ⁹ is hydrogen or C 1-4 alkyl or aryl;

n is 0, 1, 2, 3, 4, 5, 6 or 7;

ri is 0,1,2 or 3; and a is a group of formula XI or a chemical bond.

The present invention also encompasses pharmaceutically acceptable salts of the compounds of Formulas I and II.

A narrower group of compounds of Formulas I and II are compounds wherein

Q represents (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII), (XXIII), A group of formula (XXIV), (XXV), (XXVI), (XXVII), (XXVIII), (XXIX) or (XXX);

n is 0, 1, 2, 3, 4, 5, 6 or 7;

ri is 0, 1, 2 or 3;

R ⁴ is hydrogen, straight or branched or cyclic alkyl of 1 to 4 carbon atoms or alkenyl of 1 to 4 carbon atoms;

^R5 is CH, -CH _{(CH2) n-} or a bond;

R ² is hydrogen, straight or branched (C 1 -C 4) alkyl, aryl (C 1 -C 4) alkyl, or aryl;

^R1 is hydrogen, C1 -C4 alkyl or -N ^(R8) 2, -N ^(R8) SO2R ^7, -NR ⁸ CO ₂ R ^7, -NR ⁸ C (O) R ^7, -NR ⁸ C (O) N (R ⁷ ) R ⁸ ,

-N (R ⁸ ) SO 2 N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁸ ) C (O) OR ⁷ , -C (O) N (R ⁷ ) ₂ or R ⁶ ;

• ·

- R ¹⁰ is carboxyl, hydroxymethyl, tetrazolyl, acylsulfonamido, -C (O) N (R ⁷ ) ₂ , -CO ₂ R ⁹ or (VIII) or together with R ^{1 is a} cyclic group; R ¹ and R ⁶ together form a cyclic group of formula (IX) or (X);

y is oxygen or sulfur;

R ⁷ is hydrogen, or C 1-4 straight or branched, optionally substituted alkyl, straight or branched lower alkenyl, aryl (C 1-4) alkyl, substituted aryl, or 5- or 6-membered, Heteroaryl containing 1, 2 or 3 nitrogen, sulfur or oxygen atoms as heteroatoms; wherein the substituted alkyl group is substituted with hydroxy or C 1-4 alkoxy and the substituted aryl group has 1, 2 or 3 substituents selected from halogen, C 1-4 alkoxy, hydroxy or C 1-4 alkyl;

R ⁸ is hydrogen or C 1-4 straight or branched chain alkyl;

R ⁹ is hydrogen or C 1-4 alkyl or aryl;

a is a group of formula XI or a bond;

A is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; and

B is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen;

• · ·

- 11, A and B together form a condensed ring system in which adjacent carbon and nitrogen atoms are common.

For compounds of the invention, those containing an asymmetric center may exist in the form of racemates, racemic mixtures, and individual enantiomers and / or diastereomers. It will be understood by those skilled in the art that they are all within the scope of the invention.

A narrower class of compounds of formula (I) are those in which

AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH 2 - or (XXXII) - or (XXXIII) a group of the formula: in which X is nitrogen or -CR ³ , in which R ³ is cyano, -C (O) N (R ⁷ ) R ⁸ , (XXXIV), (XXXV) or (XXXVI). or a group of formula (XXXIX), (XL) or (XLI).

A further narrower group of the latter compounds is the compound of formula (I) wherein

B is a compound of formula (XLII), wherein V is nitrogen or methine and D is ethylene or -CH ₂ C (R ⁴ ) ₂ CH ₂ -.

A narrower class of compounds of formula (II) of the present invention are those in which

AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH ₂ - or (XXXII) - or

-12 (XXXIII) - in which X is nitrogen or -CR ³ , in which R ³ is cyano, -C (O) N (R ⁷ ) R ⁸ , (XXXIV) or (XXXV) ) - or a compound of formula (XL), (XLI) or (XLV), wherein in the latter, y ³ is oxygen or two hydrogen.

These compounds of formula (II) also form a separate group where a is a compound of formula (XI); and

AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH 2 - or (XXXII) or (XXXIII) a radical of formula: - wherein X represents a nitrogen atom or a CR ³ group of formula and where R ³ is cyano, -C (O) N ^{(R7) R8,} (XXXIV) or (XXXV) following group.

A narrower group of compounds of formula II are those wherein a is a bond; and

AB is a group of formula (XL), (XLI) or (XLV), and in the latter, y ³ is oxygen or two hydrogen.

Specific examples of compounds of the present invention include the following compounds and pharmaceutically acceptable salts thereof: (XLVI), (XLVII), (XLVIII), (XLIX), (L), (III), (III), ( Lili), (LIV), (LV), (LVI), (LVII), (LVIII), (LIX), (LX), (LXI), (LXll), (LXIII), (LXIV), (LXV) , (LXVI), (LXVII), (LXVIII), (LXIX), (LXX), (LXXI), (LXXII), (LXXIII), (LXXIV), (LXXV), (LXXVI I), (LXXVIII), (LXXIX) and (LXXX).

The compounds of formula (LXXXI), (LXXXI I), (LXXXIII), (LXXXIV) and (LXXXV) are also included.

The term "pharmaceutically acceptable salts" refers to the non-toxic salts of the compounds of the invention which are generally prepared by reacting the appropriate free base with a suitable organic or inorganic acid. Representative salts include: acetate, benzene sulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edizate, , glyceptate, gluconate, glutamate, glycollyl-arazanylate, hexyl resorcinate, hydrabaminate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide , methylsulfate, mycate, sun silicate, nitrate, oleate, oxalate, pamoate, palmitate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, theoclate, tosylate, triethyl.

The term "therapeutically effective amount" means an amount of an active ingredient that exerts a biological or therapeutic effect in a tissue, organ, or animal, as desired by the researching or treating physician. The term anticoagulant includes, for example, heparin and warfarin. The term thrombolytic agent includes, for example, streptokinase and tissue plasminogen activator. Platelet anti-aggregation agent includes, for example, aspirin, ticlopidine and dipyridamole.

The term alkyl means straight or branched chain alkanes, alkenes or alkynes.

The term aryl refers to 5- to 10-membered, unsaturated mono- or bicyclic ring groups.

• ·

-ΜΑ heteroaryl is a subgroup containing 1, 2, 3 or 4 heteroatoms.

The term heteroatom refers to nitrogen, oxygen or sulfur.

The term cyclic, unless otherwise specified, refers to 5 to 10 membered mono- or bicyclic saturated ring groups.

The term heterocyclic refers to ring groups containing 1,2,3 or 4 heteroatoms. In the compounds of the invention, the heteroaryl groups and the heterocyclic groups contain up to two oxygen or sulfur atoms.

The term "alkoxy" refers to groups containing the alkyl moiety as defined above.

Arylalkyl and alkylaryl groups are understood to mean those alkyl groups as defined above and aryl groups as above. In the case of Co _n or Cl _n , n is a number from 1 to 10 or 2 to 10 or refers to the alkyl portion of arylalkyl or alkylaryl groups.

The term halogen means fluorine, chlorine, iodine and bromine.

The term oxy refers to an oxygen atom. By oxo is meant (= 0) a group. The term thio refers to a sulfur atom. In the nomenclature used herein, the terminal end of a particular side chain is mentioned first, and then the related functional group, such as C 1-6 alkyl substituted with C 1-6 alkylcarbonylamino, corresponds to formula LXXXVI.

The following abbreviations are used for the various reagents used in the preparation procedures and examples which follow:

· .. · · '· *' ·:: ·· ·· ···· .. · .. · - 15- BOC (Boc): tert-butoxycarbonyl Pd-C: palladium on activated carbon DMF: dimethylformamide DMSO: dimethylsulfoxide CBZ: benzyloxy carbonyl CH ₂ CI ₂ : methylene chloride CHCI ₃ : chloroform EtOH: ethanol MeOH: methanol EtOAc: ethyl acetate HOAc: acetic acid BOP: benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexa Fluoro-phosphate EDC: 1- (3-dimethyl-aminopropyl) -3-ethylcarbodiimide Oxone: potassium peroxymonosulfate LDA: lithium diisopropylamide DMA: N, N-dimethylaniline HOBT: hydroxybenzotriazole.

The compounds of the present invention are useful in the so-called "cell adhesion" activity. inhibition of integrin protein complex function. The compounds of the invention may also be administered to patients in whom inhibition of human or mammalian platelet aggregation or adhesion is desired.

Some of the compounds of the invention are rapidly cleared from the circulation and are particularly effective in inhibiting platelet aggregation. so these

Compounds of -16a may be used in peripheral arterial surgery (arterial transplantation or carotid artery surgery) and coronary artery surgery, in which manipulation of arteries and organs and / or interactions between arterial surfaces and platelets results in platelet aggregation. Aggregated platelets can cause thrombi and thromboembolism (i.e., the compounds of the present invention may be administered to patients undergoing such surgical intervention to prevent the formation of thrombi and thromboembolism).

The compounds of the present invention may be administered in the form of conventional oral formulations such as tablets, capsules (all of which may be formulated as sustained release or delayed release), pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. The compounds of the present invention may also be administered intravenously (by bolus injection or infusion), intraperitoneally, subcutaneously, sublingually, intranasally or intramuscularly, and methods of administration and related formulations are well known to those skilled in the art. An effective amount of the compound of the present invention as an anti-aggregation agent is required but not yet toxic.

The compounds of the present invention can be administered to patients in whom prevention of thrombosis is desirable by inhibiting fibrinogen at the platelet membrane-glycoprotein complex IIb / IIIa receptor. Thus, these compounds can be used in peripheral arterial surgery (arterial transplantation or cervical vascular surgery) and in coronary artery surgery, where manipulation of arteries and organs and / or interference between arterial surfaces and platelets occurs.

interaction leads to platelet aggregation. Aggregated platelets can cause thrombi and thromboembolism. Thus, the compounds of the present invention can be administered to patients undergoing such surgical intervention to prevent the formation of thrombi and thromboembolism.

In coronary heart surgery, extracorporeal circulation is routinely used to enrich the blood with oxygen. Platelets adhere to the surfaces of the body outside the body. Adhesion depends on the interaction between gpl1b / IIIa points on platelet membranes and fibrinogen adsorbed to the surface of the extracellular unit (Gluszko et al., Amer. J. Physiol., 252 (H), 615-621 (1987)). Platelets separated from artificial surfaces show impaired haemostatic function. The compounds of the invention may be used to prevent said adhesion.

Further use of the compounds of the invention is in the prevention of platelet thrombosis, thromboembolism and reocclusion during and after thrombolytic therapy, and in the prevention of platelet thrombosis, thromboembolism and reocclusion in vascular surgery or in coronary artery bypass graft and other arterial bypass. The compounds of the invention may also be used for the prevention of myocardial infarction.

The mode of administration of the compounds of the invention will depend on a number of factors including the nature, species, age, weight, sex and health of the patient; the severity of the disease to be treated; the route of administration; the patient's renal and hepatic function; and depends on the nature of the compound or salt used. A physician or veterinarian skilled in the art can determine or prescribe the active ingredient

-18 which is capable of interfering with, curing or preventing the course of the disease being treated.

Oral dosages of the compounds of the invention, when used in the above-mentioned areas of activity, are about 0.01 mg to 100 mg, preferably 0.05 to 100 mg, especially 0.1 to 20 mg per kg body weight per day. Intravenously, the most preferred dose at a constant infusion flow rate is about 1-10 pg / kg / min. Preferably, the compounds of the invention are administered in divided doses of 2, 3 or 4 times daily. In addition, the preferred compounds of the invention may be administered intranasally topically using suitable intranasal vehicles or transdermally in the form of transdermal patches well known to those skilled in the art. For transdermal administration, it will be appreciated that the dose is continuous, rather than a single dose.

Pharmaceutical compositions containing the compounds of the present invention typically contain, in addition to the active ingredient, customary pharmaceutical excipients, diluents or other excipients, hereinafter collectively referred to as excipients. The choice of carrier will depend on the intended route of administration, for example, the use of oral tablets, capsules, elixirs or syrups, and will be made according to principles well known in the art of pharmacy.

For example, in the case of tablets or capsules for oral administration, the active ingredient may be formulated with an oral, non-toxic, pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate. :, ·· in combination with nitt or sorbitol; in the case of oral administration in liquid form, the active ingredient may be combined with any non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol or water for oral administration. In addition, suitable binders, lubricants, disintegrating agents and coloring agents may be added to the mixture as desired or necessary. Suitable binders include starch, gelatin, natural sugars such as glucose or β-lactose, sweeteners based on cereals, natural and synthetic resins such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol. and waxes. Examples of lubricants that may be used in such formulations include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride. Examples of disintegrants include starches, methylcellulose, agar, bentonite, and xanthan gum.

The compounds of the invention may also be administered by the so-called in the form of liposome delivery systems such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed with various phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

The compounds of the invention may be administered using monoclonal antibodies, in which the molecules of the compounds are coupled thereto. The compounds of the invention may also be coupled with soluble polymers as active ingredient carriers. Examples of such polymers include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethyl ·

-20-acrylamide phenol, polyhydroxyethyl aspartamide phenol or polyethylene oxide polylysine substituted with palmitoyl residues. The compounds of the invention may also be linked to biodegradable polymers, such as copolymers of polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, polyepsilone caprolactone, polyhydroxy butyric acid, polycarboxylate, hydrogel-block copolymers.

However, the compounds of the invention may be co-administered with suitable anticoagulants. Such anticoagulants include agents that inhibit platelet aggregation, such as heparin, aspirin, warfarin, dipyridamole and other compounds, as well as known substances capable of inhibiting blood clot formation, as well as thrombolytic agents such as plasminogen activators or thrombolytic agents. By co-administration, various vascular pathological symptoms are preferably treated.

The compounds of the invention may be prepared by the following examples. All of the most preferred compounds of the invention are described in these Examples. However, these compounds are not meant to be the only compounds that can be produced according to the invention, and these compounds or portions thereof can be combined arbitrarily to give a gene. The following examples, therefore, illustrate the process of the invention. One skilled in the art will appreciate that reaction conditions other than the preparative examples described below may be used. Temperatures in the examples are values in ° C unless otherwise indicated.

···· ··

• * ···· · * 'I ··

In addition to the preparation examples set forth below, many of the compounds of the invention exhibit in vitro bioactivity. To this end, one of the assays used is the evaluation of the fibrinogen receptor antagonist activity by measuring the inhibition of ADP-stimulated platelets. Aggregation requires fibrinogen to bind to and occupy the platelet fibrinogen receptor site. Inhibitors of fibrinogen inhibit this aggregation. In the ADP-stimulated platelet aggregation assay (performed to determine the inhibitory ability of the compounds of the invention), human platelets were isolated from fresh blood by collecting platelets in acid citrate / dextrose, followed by differential centrifugation, and then labeled with Sepharose 2B. divalent deionized Tyrode buffer (pH 7.4) containing 2% bovine serum albumin.

Platelet aggregation was performed at 37 ° C on a Chronolog aggregometer. The reaction mixture contained 2 x 10 ⁸ / ml of gel-filtered human platelets, 100 pg / ml of fibrinogen, 1 mmol of calcium ions and the test compound. Aggregation was achieved by adding 10 mM ADP 1 minute after the other components were added. The reaction is then allowed to proceed for at least two minutes. The degree of inhibition of aggregation is expressed as the percentage of aggregation measured in the absence of inhibitor. The IC50 value is the dose of a given compound that reduces aggregation by 50% compared to a control in the absence of a test compound.

Scheme A illustrates the preparation of sulfonamide derivatives as intermediates.

-22 • ♦. . · * · ··· ·····. .. ·. ·, ·, * ·

L-Asparagine-a-Butanesulfonamide (also known as N- (n-Butylsulfonyl) -L-Asparagine) - A-2

A solution of 6.45 g (48.9 mmol) of L-asparagine and 2.0 g (50.0 mmol) of sodium hydroxide in 100 ml of 50% aqueous dioxane was cooled to 0 ° C in an ice bath. a solution of sodium hydroxide (2.2 g, 55.0 mmol) in water (50 mL) and butanesulfonyl chloride (7.0 mL, 53.9 mmol) were added under vigorous stirring for 30 minutes. alternate reagents are added. After the addition was complete, the reaction mixture was concentrated under reduced pressure to a volume of 50 ml. The resulting aqueous residue was cooled, acidified with concentrated hydrochloric acid and then extracted three times with 100 ml of ethyl acetate. The organic extracts were dried over anhydrous sodium sulfate and concentrated to a volume of approximately 50 ml. Anhydrous diethyl ether (50 mL) was added to the concentrate and the white precipitate was collected by vacuum filtration to give compound A-2, m.p. 154-155 ° C.

L-Boc-α-butanesulfonamido-β-amino-alanine [aka 2 (S) - (n-butylsulfonylamino) -3- (N-Boc-amino) -propionic acid] - (A- A solution of sodium hydroxide (6.04 g, 151 mmol) in water (50 mL) was cooled to 0 ° C and elemental bromine (1.40 mL, 26.9 mmol) was added. The resulting solution was stirred at 0 ° C for 5 min and then 5.23 g (20.7 mmol) of A-2 and 1.66 g (41.4 mmol) of sodium hydroxide in 15 mL were added all at once. water-cooled solution. The resulting reaction mixture was stirred at 0 ° C for 5 min and then warmed to 80 ° C over 15 min. The solution was cooled to 25 ° C and acidified with 11 ml of 12N hydrochloric acid. Stirring is then continued until gas evolution subsides. Then the solution

After basification by addition of N sodium hydroxide solution, tetrahydrofuran (20 ml) and di-tert-butyl dicarbonate (9.0 g, 41.4 mmol) were added. After stirring at 25 ° C overnight, the tetrahydrofuran is removed under reduced pressure, and the remaining basic aqueous phase is extracted twice with 50 ml of ethyl acetate each time. The aqueous phase was then made basic with 10% potassium bisulfate solution and extracted with ethyl acetate (3 x 100 mL). The combined acidic extracts were dried over anhydrous sodium sulfate, filtered, and evaporated to give A-2 as a white solid, m.p. 111-112 ° C.

The compound of formula (A-4) can be prepared as shown in Scheme B.

2 (S) - (n-Butylsulfonylamino) -3-aminopropionic acid (A-4)

A solution of A-3 (3.83 g, 11.8 mmol) in ethyl acetate (200 mL) was cooled to 0 ° C and hydrogen chloride gas was bubbled through the solution for 5 min. The solution was then warmed to 25 ° C, stirred for 30 minutes, then concentrated under reduced pressure to 50% of its original volume and diluted with 100 ml of diethyl ether. The resulting white precipitate was collected by vacuum injection to give A-4.

Ethyl 2 (S) - (n-butylsulfonylamino) -3-aminopropionate (A-5)

A solution of the compound (A-4) (1.0 g, 3.8 mmol) in anhydrous ethanol (50 mL) was saturated with gaseous hydrogen chloride and refluxed for 3 hours. The solvent is then evaporated to give pure A-5 as a white precipitate.

¹ H NMR (300 MHz, CD ₃ , OD): δ 4.38 (m, 2H), 4.32 (m, 1H), 4.23 (m, 2H), 2.85 (m, 2H) , 1.65-1.45 (m, 4H), 1.3 (t, 2H), 0.96 (t, 3H).

The following compounds of formulas (A-6) to (A-8) may be prepared as shown in Scheme C.

N-Tosyl-L-Asparagine - A-6

To a 500 ml round-bottomed flask equipped with a magnetic stir bar and a addition funnel was added 10.0 g (75.7 mmol) of L-asparagine and 85 ml (1.1 molar equivalent) of 1 N sodium hydroxide solution was added. Meanwhile, 15.88 g (83.27 mmol) of p-toluenesulfonyl chloride was dissolved in 100 mL of ethyl acetate and the resulting solution was added to the reaction vessel with vigorous stirring. Then, 85 ml (1.1 molar equivalent) of 1 N sodium hydroxide solution was added to the addition funnel and this solution was added dropwise over 2 hours with vigorous stirring. The reaction mixture was then stirred at room temperature for an additional 2 hours, then the organic and aqueous layers were separated and the aqueous layer was washed twice with 50 mL each of ethyl acetate. The aqueous phase is then cooled to 0 ° C and then acidified with concentrated hydrochloric acid. A white crystalline precipitate is obtained. Recrystallization from hot water gives the compound (A-6).

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 7.91 (d, J = 8.79 Hz, 1H), 7.64 (d, J = 8.06 Hz, 2H), 7.32 [s, d (overlapping), J = 8.06 Hz, 3H], 6.87 (s, br, 1H), 4.03 (m, 1H), 3.32 (s, H ₂ O); 2.49 (m, 1H), 2.43 (d, d, J = 7.08, 15.38 Hz, 1H), 2.35 (s, 3H), 2.21 (d, d, J = 6.11, 15.38 Hz, 1H).

-252 (S) - (Tosylamino) -3-aminopropionic acid - A-7

A solution of sodium hydroxide (22.0 g, 550 mmol) in water (100 mL) was cooled to 0 ° C and elemental bromine (5.03 mL, 97.5 mmol) was added dropwise. The resulting solution was then stirred at 0 ° C for 10 min, and 21.5 g (75.0 mmol) of A-6 and 6.68 g (161 mmol) of sodium hydroxide in 50 mL were added in one portion. water, cooled to 0 ° C. After stirring at 0 ° C for 20 minutes, the reaction mixture was heated to 80 ° C over 30 minutes and then cooled. The cooled solution was adjusted to pH 7 with concentrated hydrochloric acid and the white precipitate formed was filtered off to give A-7.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.2-7.2 [broad, 2H, (NH.COOH)], 7.70 (d,

J = 8.18 Hz, 2H), 7.38 (J = 8.18 Hz, 2H), 3.7 -3.0 [broad, 2H, (NH ₂ , H ₂ O) J, 3.12 ( q, J = 4.76 Hz, 1H), 2.99 (d, d, J = 4.64, 11.96 Hz, 1H), 2.79 (d, d, J = 9.52, 11, 96 Hz, 1H), 2.36 (s, 3H).

Tert-Butyl 2 (S) - (toluenesulfonylamino) -3-aminopropionate (A-8) in a liter of pressurized vessel was treated with 5.0 g (19.4 mmol) of dioxane (100 mL). The compound of formula A-7 is suspended and the vessel is cooled to -15 ° C and then 100 ml of isobutylene are condensed in dioxane. Concentrated sulfuric acid (5 ml) was added and the vessel was sealed and stirred at room temperature for 36 hours. The vessel is then opened and the excess isobutylene is carefully drained. The reaction mixture was diluted with ethyl acetate (200 mL) and washed with 1N sodium hydroxide solution (200 mL). The organic layer was then dried over anhydrous sodium sulfate, filtered and evaporated. A white crystalline precipitate was obtained which gave A-8.

-26 ^1H-NMR (300 MHz, _CDCl3): δ 7.68 (d, J = 8.18 Hz, 2H), 7.35 (d, J = 8.18 Hz, 2H);

3.85 (m, H), 2.93-2.79 (m, 2H), 2.32 (s, 3H), 1.38 (s, 9H).

The preparation of compounds (1-2) and (1-3) is illustrated in Scheme 1.

Tert-Butyl 2-hydrazinopyridine-3-carboxylate - (1-2)

A solution of tert-butyl 6-chloronicotinate (735 g, 3.44 mmol) in ethanol (5 mL) was cooled to 0 ° C and a solution of anhydrous hydrazine (2.75 g, 86 mmol) in ethanol (5 mL) was added. The resulting reaction mixture was allowed to warm to 25 ° C and was then stirred at this temperature for 6 hours and then at 60 ° C for 2 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with water and then with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give (1-2) as an oil, which was used directly in the next step.

Tert-Butyl 2,3-dihydro-3-oxo-1,4-triazolo [4,3-a] pyridin-6-ylcarboxylate Compound 1-3

To a solution of the compound (1-2) in toluene (35 mL) was slowly added a solution of bis (trichloromethyl) carbonate (1.15 g, 3.9 mmol) in toluene (35 mL) heated to reflux, and the resulting mixture was stirred at room temperature. reflux for 1.2 hours and then cool. Water was added and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate, evaporated and the residue subjected to flash chromatography on silica gel to give compound (1-3).

-27 ^1H-NMR (300 MHz, _DMSO-d6): δ 1.58 (s, 9H), 7.25 (d, 1H), 7.45 (d, 1H), 8.25 (s, 1H), 12.40 (s, 1H).

The preparation of compound (1-5) is shown in Scheme 2 and the preparation of compound (1-4) is shown in Scheme 3.

2- (N-CBZ-Piperidin-4-yl) ethyl iodide - Compound 1-4

To a mixture of 4- (2-hydroxyethyl) piperidine (5.0 g, 38.7 mmol) from Aldrich, saturated aqueous sodium bicarbonate (50 mL) and dichloromethane (150 mL) was added 6.05 mL (42.5 mmol) benzyl chloroformate and stirred at room temperature for 2.5 hours. The organic layer was then separated, washed with water and brine, dried over anhydrous sodium sulfate, filtered and evaporated to give the protected alcohol as a colorless oil.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.26 (m, 5H), 5.13 (s, 2H), 4.23 (d, 2H), 3.63 (t,

2H), 2.85 (t, 2H), 1.83 (d, 2H), 1.68 (m, 2H), 1.43 (m, 1H), 1.03 (m, 2H).

To this 6.3 g (23.9 mmol) of this protected alcohol is added 7.0 g (23.9 mmol) of Ph ₃ P, 6.06 g (23.9 mmol) of elemental iodine and 1.95 g (28). 7 mmol) of imidazole in 100 ml of benzene and the resulting mixture was refluxed for 2.5 hours. The reaction mixture was cooled, filtered and evaporated. The residue was purified by chromatography on silica gel, eluting with ethyl acetate / hexane (1: 1). This gives a compound of formula (I-4) as a white precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.26 (m, 5H), 5.14 (s, 2H), 4.23 (d, 2H), 3.86 (m,

4H), 1.85 (d, 2H), 1.68 (m, 2H), 1.45 (m, 1H), 1.03 (m, 2H).

3 - [(2,3-Dihydro-3-oxo- (2- (N-CBZ-piperidin-4-yl) ethyl) -1,2,4-triazolo [4,3-a] pyridine-6) -yl] carboxylic acid - a compound of the formula (I-5)

To a solution of 1-3 (350 mg, 1.6 mmol) in acetonitrile (20 mL) was added potassium carbonate (630 mg) in powder form and the resulting mixture was heated at 60 ° C for 20 h. The mixture was cooled to 25 ° C, water was added and the aqueous mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel flash chromatography using 35% ethyl acetate / hexane as eluent. This gives the tert-butyl ester corresponding to the desired compound as a yellow oil.

This crude tert-butyl ester is then converted to the corresponding acid by addition of a mixture of 15 ml of methylene chloride and 15 ml of trifluoroacetic acid at 0 ° C and then heated to 25 ° C over a period of 1.2 hours. . The reaction mixture was evaporated to dryness in vacuo, water was added to the residue, and the aqueous mixture was extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and evaporated. The residue was crystallized from ethyl acetate: diethyl ether (25: 1). The title compound (1-5) is thus obtained as a pale yellow powder.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 1.05 (m, 2H), 1.45 (m, 1H), 1.75 (m, 4H), 2.79 (m, 2H) , 3.95 (q, 4H), 5.05 (s, 2H), 7.30 (m, 6H), 7.50 (d, 1H), 8.25 (s, 1H).

The compound of formula (1-6) can be prepared as shown in Scheme 4.

-293 - / ((2,3-Dihydro-3-oxo-2- [2- (N-CBZ-piperidin-4-yl) ethyl] -1,2,4-triazolo [4,3-a] Pyridin-6-yl} carbonyl] -amino-propionic acid (1-6) To a solution of the compound (1-5) in mg (0.18 mmol) in 1 mL of dimethylformamide was added successively 40 mg (0 , 26 mmol) hydroxybenzotriazole, 87 µl (0.5 mmol) diisopropylethylamine, 40 mg (0.26 mmol) ethyl 2-aminopropionate hydrochloride and 50 mg (0.26 mmol) EDC The reaction mixture was stirred at 25 ° C for 15 hours, then dissolved in water and extracted with ethyl acetate, the organic phase was washed with water, brine, dried over anhydrous sodium sulfate, and dried over anhydrous sodium sulfate. evaporated to give the corresponding ethyl ester of the desired compound.

This crude ethyl ester was dissolved in tetrahydrofuran (5 mL) and treated with water (5 mL) followed by 1N aqueous lithium hydroxide (0.37 mL). The reaction mixture was stirred at 25 ° C for 3 hours, then dissolved in water, followed by extraction with ethyl acetate. The extract was washed with water followed by saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give compound 1-6, m.p. 201-203 ° C.

The preparation of compound (1-7) is illustrated in Scheme 5.

3 - / (((2,3-Dihydro-3-oxo-2- (2-piperidin-4-yl) ethyl] -1,2,4-triazolo (4,3-a] pyridin-6-yl } -carbonyl] -amino-propionic acid (1-7) To a solution of the compound (1-6) in mg (0.129 mmol) in acetonitrile (15 mL) at 0 ° C was added 107.0 mg (0.533 mmol) of iodine. trimethylsilane and the resulting mixture was stirred for 0.5 hour, quenched with water, extracted with diethyl ether and silica gel.

The residue was purified by flash chromatography using a 10: 1: 1 by volume mixture of ethanol, ammonium hydroxide solution and water. On evaporation, a white foam is obtained. Crystallization from ethanol gives compound (1-7) as a white solid, m.p. 267-269 ° C.

The preparation of compound (1-9) is illustrated in Scheme 6. 2 (S) - [6- (butylsulfonyl) amino] -3 - / [{2,3-dihydro-3-oxo-2- / 2- (N-CBZ-piperidin-4-yl) ethyl (-1,2,4-Triazolo [4,3-a] pyridin-6-yl} carbonyl] amino] -propionic acid (1-8)

As described for the preparation of the compound (1-6), the compound (1-5) is coupled with the compound (A-4) to give the compound (1-8).

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.26 (t, 1H), 8.50 (s, 1H), 7.63 (d, 1H), 7.54 (d, 1H) , 7.4-7.31 (m, 6H), 5.01 (s, 2H), 4.10 (m, 1H), 3.96 (m, 4H), 3.60 (m, 1H), 3.46 (m, 1H), 2.95 (t, 2H), 2.73 (broad m, 2H), 1.71 (m, 2H),

1.53 (m, 2H), 1.46 (m, 1H), 1.01 (m, 2H), 0.83 (t, 3H).

2 (S) - [(n-butylsulfonyl) amino] -3 - / [{2,3-dihydro-3-oxo-2- / 2- (piperidin-4-yl) ethyl / -1, 2,4-Triazolo [4,3-a] pyridin-6-yl} carbonyl] -amino] -propionic acid (1-9)

Treatment of the compound (1-8) with trimethylsilyl iodide in acetonitrile as described for the preparation of the compound (1-7) gives the compound (1-9).

¹ H-NMR (300 MHz, D ₂ O): δ 8.10 (s, 1H), 7.43 (d, 1H), 7.15 (d, 1H), 4.00 (m, 4H), 3.60 (d, 2H), 3.31 (m, 2H), 3.23 (d, 2H), 3.93 (m, 2H), 3.81 (t, 2H), 1.93 (d) , 2H), 1.80 (m, 2H), 1.6-1.23 (m, 7H), 0.85 (t, 3H).

The preparation of compound (2-3) is illustrated in Scheme 7.

Methyl 6-methylpyridine-3-carboxylate - 2-2

Into a 250 ml three-necked flask fitted with a dropping funnel, a vertical reflux condenser and a calcium chloride drying tube, a solution of 5 g (36.5 mmol) of 6-methylnicotinic acid, i.e. 2-1, in 100 ml of anhydrous methanol, the solution was cooled to -15 ° C in an ice-acetone bath and 5 mL (69.1 mmol) of SOCl ₂ was added dropwise. The solution was then refluxed for 3 hours, cooled and the solvent removed under reduced pressure. To the residual white precipitate was added 60 mL of saturated aqueous sodium bicarbonate solution, followed by extraction with methane (3 x 50 mL). The combined extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting oil was allowed to crystallize on standing to give 2-2 as a white precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 9.05 (d, J = 1.4 Hz, 1H), 8.08 (dd, J = 1.4 and

6.8 Hz), 7.19 (d, J = 6.8 Hz, 1H), 3.98 (s, 3H), 2.63 (s, 3H).

Methyl 6- (bromoethyl) pyridine-3-carboxylate - (2-3)

To 10.6 g (71.5 mmol) of compound 2-2 is added 12.73 g (71.5 mmol) of NBS, 100 mg of benzoyl peroxide and 200 mL of carbon tetrachloride, and then under reflux under an inert atmosphere Heat to reflux for 18 hours. The reaction mixture was cooled, filtered and concentrated, and the resulting viscous orange oil was subjected to flash chromatography on silica gel using 20% ethyl acetate / hexane as eluent. This gives the desired compound of formula (2-3).

• · · • ·

-32 ^1H-NMR (300 MHz, _CDCl3): δ 9.05 (d, J = 1.4 Hz, 1H), 8.08 (dd, J = 1.4 and

6.8 Hz), 7.19 (d, J = 6.8 Hz, 1H), 5.38 (s, 2H), 3.98 (s, 3H).

The preparation of compound (2-6) is illustrated in Scheme 8.

Methyl 6- [2- (N-Boc-piperidin-4-yl) -ethylamino] -methyl-piperidine-3-carboxylate - 2-5

1.0 g (4.34 mmol) of compound (2-3), 2.16 g (10.0 mmol) of compound (2-4) and 0.66 g (4.4 mmol) of potassium carbonate 100 of anhydrous acetonitrile was weighed into a 250 ml reaction vessel and refluxed for 3 hours. After cooling and filtration, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel, eluting with 10% methanol in ethyl acetate. The residue (2-5) is obtained as a yellow residue. ¹ H-NMR (CDCl 3): δ 9.18 (d, J = 1.4 Hz, 1H), 8.15 (dd, J = 1.4 and 6.8 Hz, 1H), 7.39 (d , J = 6.8 Hz, 1H), 4.08 (broad d, J = 12 Hz, 2H), 3.98 (s, 2H), 3.95 (s, 3H), 2.75 (overlapping m , 6H), 1.78 (d, J = 12 Hz, 2H), 1.5 (overlapping m, 4H),

1.4 (s, 9H), 0.98 (m, 2H).

Methyl 1- (chlorocarbonyl) -2,3-dihydro-3-oxo-2- [2- (N-Boc-piperidin-4-yl) -ethyl] -imidazo [1,5-a] pyridine. 6-yl Carboxylate 2-6

To a solution of 2-5 (480 mg, 1.27 mmol) in toluene (50 mL) was added Ν, Ν-dimethylaniline (645 mL, 4.08 mmol) and the resulting mixture was cooled to 0 ° C. A solution of triphosgene (1.13 g, 3.18 mmol) in toluene (15 mL) was added dropwise over 30 min. The resulting reaction mixture was then heated at 100 ° C for 1.5 hours and then cooled, washed twice with 1 N hydrochloric acid, once with water and once with saturated aqueous sodium chloride (using 50-50 mL each).

It was dried over anhydrous sodium sulfate and evaporated. 515 mg of the compound of formula (2-6) are obtained as a yellow crystalline precipitate. ¹ H-NMR (CDCl 3): δ 8.82 (d, J = 1.4 Hz, 1H), 8.25 (d, J = 6.8 Hz, 1H), 7.91 (d,

J = 6.8 Hz, 1H) (dd, J = 1.4 and 6.8 Hz, 1H), 4.08 (broad d, J = 12 Hz, 2H),

3.98 (s, 2H), 3.95 (s, 3H), 2.75 (overlapping m, 6H), 1.78 (d, J = 12 Hz, 2H),

1.5 (overlapping m, 4H), 1.4 (s, 9H), 0.98 (m, 2H).

The preparation of compound (2-7) is illustrated in Scheme 9.

Methyl 1 - [(N, N-diethylamino) carbonyl] -2,3-dihydro-3-oxo-2- [2- (N-Boc-piperidin-4-yl) ethyl] imidazo [ 1,5-a] pyridin-6-ylcarboxylate - 2-7

To a solution of compound (2-6) (0.3 g, 0.64 mmol) in dichloromethane (100 mL) was added diethylamine hydrochloride (105 mg, 0.97 mmol) and saturated aqueous sodium bicarbonate (50 mL). and the resulting biphasic mixture was stirred for 1 hour, then the organic phase was separated, washed with 10% citric acid solution, then with 50 ml of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to a residue. 2-7).

¹ H-NMR (CDCl 3): δ 8.45 (d, J = 1.4 Hz, 1H), 7.15 (dd, J = 1.4 and 6.8 Hz, 1H), 6.78 (d, J = 6.8 Hz, 1H), 4.08 (m, 4H), 3.95 (s, 3H), 3.51 (m, 4H), 2.75 (m, 3H),

1.78 (d, J = 12 Hz, 2H), 1.5 (overlapping m, 4H), 1.4 (s, 9H), 1.1 (t, 6H), 0.98 (m, 2H) .

The compound of formula (2-9) can be prepared as shown in Scheme 10.

• · ·

-34-tert-Butyl 1 - [[(N, N-diethylamino) carbonyl] -2,3-dihydro-3-oxo-2- [2- (N-Boc-pipendin-4-yl) ethyl] -imidazo [1,5-a] pyridin-6-yl] carbonyl-aminopropionate - 2-8

To a solution of compound 2-7 (315 mg, 0.64 mmol) in methanol (10 mL) was added water (15 mL) and 1N sodium hydroxide solution (0.725 mL), and the resulting mixture was stirred at room temperature for 3.5 hours. The organic solvent was then removed under reduced pressure and the aqueous residue was acidified with citric acid and extracted several times with dichloromethane. The organic extracts were washed with water followed by brine, dried over anhydrous sodium sulfate, filtered and evaporated to give the desired acid.

¹ H-NMR (CDCl 3): δ 8.32 (d, J = 1.4 Hz, 1H), 7.10 (dd, J = 1.4 and 6.8 Hz, 1H), 6.78 (d , J = .8Hz, 1H), 4.08 (m, 4H), 3.51 (m, 4H), 2.75 (m, 3H), 1.78 (d,

J = 12 Hz, 2H), 1.5 (overlapping m, 4H), 1.4 (s, 9H), 1.1 (t, 6H), 0.98 (m, 2H).

105 mg (0.215 mmol) of this acid is dissolved in 10 mL of dichloromethane and then 48 mL (0.47 mmol) of triethylamine are added and the resulting mixture is cooled to -10 ° C. Isobutyl chloroformate (30 mL, 2.36 mmol) was added and the resulting mixture was stirred at -10 ° C for 30 min. A solution of 58.7 mg (0.323 mmol) of β-alanine tert-butyl ester hydrochloride and 32 mL (0.323 mmol) of triethylamine in 10 mL of methylene chloride was added and the resulting solution was warmed to room temperature. The reaction mixture is then washed with 10-10 ml of 10% aqueous citric acid, water and saturated brine, dried over anhydrous sodium sulfate, concentrated and chromatographed to give compound (2-8).

-35 ^1H-NMR (CDCl₃) δ 8.16 (s, 1H), 7.02 (d, J = 6.8 Hz, 1H), 6.78 (d, J = 6.8 Hz, 1H )

6.63 (t, J = 5.6 Hz, 1H), 4.08 (m, 4H), 3.51 (m, 6H), 2.75 (m, 2H), 2.45 (m, 2H) ), 1.78 (d, J = 12 Hz, 2H), 1.5 (overlapping m, 6H), 1.4 (s, 9H), 1.37 (s, 9H), 1.1 (t, 6H), 0.98 (m, 2H).

- [(N, N-Diethylamino) carbonyl] -2,3-dihydro-3-oxo-2 - [(2- (piperidin-4-yl) ethyl} imidazo [1,5-a] ] pyridin-6-yl] carbonyl / propionic acid - 2-9

A solution of 2-8 (100 mg, 0.16 mmol) in ethyl acetate (20 mL) was stirred at 0 ° C for 5 min under anhydrous hydrogen chloride gas and stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure, and the residue was triturated with ethyl acetate and filtered to give 2-9.

¹ H-NMR (DMSO-d ₆ ): δ 8.90 (bs, 1H), 8.60 (bs, 1H), 8.3 (s, 1H), 7.6 (t, 3H), 7.1 (d, 1H), 6.89 (d, 1H), 4.08 (m, 4H), 3.51 (m, 6H), 2.75 (m,

2H), 2.45 (m, 2H), 1.78 (d, J = 12 Hz, 2H), 1.5 (overlapping m, 6H), 1.1 (t,

6H), 0.98 (m, 2H).

The compound of formula (2-4) can be prepared as shown in Scheme 11.

2- (N-Boc-piperidin-4-yl) -ethanol - Compound 2-11

A solution of 130 g (1.0 mole) of 4-piperidin-2-ethanol (2-10) available from Aldrich in 700 ml of dioxane is cooled to 0 ° C and 336 ml (1.0 mole) of 3 N are added. sodium hydroxide solution and 221.8 g (1.0 mole) of di-tert-butyl dicarbonate. The ice bath was then removed and the reaction mixture was stirred overnight. The reaction mixture was evaporated, diluted with water and extracted several times with diethyl ether. Unified Foods · V · · · · · ·

The phases were washed with a saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to give 2-11.

TLC Rf = 0.37 using ethyl acetate: hexanes (1: 1 v / v) as the eluent and staining with ninhydrin.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 4.07 (bs, 2H), 3.7 (bs, 2H), 2.7 (t

J = 12.5 Hz, 2H), 1.8-1.6 (m, 6H), 1.51 (s, 9H), 1.1 (ddd, J = 4.3, 12.5, 12 Hz) , 2 H).

2- (N-Boc-piperidin-4-yl) -ethyl-iodide - Compound 2-12

To a solution of the compound 2-11 (10.42 g, 0.048 mol) in benzene (400 ml) was added imidazole (4.66 g, 0.068 mol), triphenylphosphine (15.24 g, 0.05 mol) and elemental iodine (0.048 mol). . After 6 hours, the reaction mixture was filtered and the filtrate was evaporated to give a dark residue. This was then purified by flash chromatography on silica gel eluting with 10% ethyl acetate in hexane to give 2-12 as a yellow oil.

2- (N-Boc-pipendin-4-yl) -ethylamine - Compound (2-4)

To a solution of 27.9 g (0.082 mol) of 2-12 in 400 ml of dimethyl sulfoxide was added 5.01 g (0.086 mol) of sodium azide at room temperature and the resulting solution was heated at 65 ° C for 2 hours. . After cooling, the reaction mixture was diluted with ethyl acetate (250 mL), extracted with water (2 x 100 mL), and brine (50 mL, 50 mL). After drying over magnesium sulfate and removal of the solvent, the desired azide is obtained as a yellow oil having a Rf value of 0.5 on silica gel, eluting with 70% acetone in hexane.

To a solution of this azide (19.3 g, 0.076 mol) in tetrahydrofuran (400 ml) and water (195 ml) was added triphenylphosphine (80.0 g, 0.305 mol) at room temperature and the resulting mixture was stirred at room temperature for 3 hours. The organic solvents are then removed in vacuo and the residue is adjusted to pH 2 with 10% potassium bisulfate. The mixture was extracted with ethyl acetate (4 x 100 mL) and the combined extracts (2 x 100 mL x 10% potassium bisulfate). The combined aqueous phase was adjusted to pH 10 with 2N sodium hydroxide solution and extracted with dichloromethane (4 x 200 mL). The combined extracts were dried over magnesium sulfate and the solvent removed to give 24 as an oil, Rf 0.3 on silica gel using 10% methanol in chloroform in ammonia.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 4.05 (broad, 2H), 2.72 (t, J = 7.2 Hz, 2H), 2.62 (m, 2H), 1.64. (d, J = 12.2 Hz, 2H), 1.43 (s, 9H), 1.42-1.32 (m, 5H), 1.09 (m, 2H).

The preparation of compound (3-2) is illustrated in Scheme 12.

A solution of dimethyl 1- (2-bromoethyl) pyrazole-3,5-dicarboxylate 3-2 in pyrazole-3,5-dicarboxylic acid (431 mmol) in anhydrous methanol (1 L) was treated with anhydrous hydrogen chloride gas. After 30 minutes, the reaction mixture was allowed to cool to room temperature and allowed to stand for 16 hours. The solution was then refluxed for 3 hours, cooled and the solvent removed under reduced pressure. The received

* · · · · ·· ···· «·

Saturated sodium bicarbonate solution (600 mL) was added to the white precipitate, followed by extraction with dichloromethane (3 x 500 mL). The combined extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting white precipitate is then recrystallized from methanol with addition of anhydrous diethyl ether to give dimethyl pyrazole-3,5-dicarboxylate, (3-1a). ¹ H-NMR (CDCl 3): δ 7.38 (s, 1H), 3.98 (s, 3H), 3.93 (s, 3H).

To a solution of the ester thus obtained in 5.0 g (27.2 mmol) of anhydrous acetonitrile was added 5.2 g (40.0 mmol) of potassium carbonate and 25.0 ml (291 mmol) of 1,2-dibromoethane. and the resulting reaction mixture is refluxed under an argon atmosphere. After refluxing for 25 minutes, the resulting suspension is cooled, filtered and the filtrate is evaporated to dryness under reduced pressure. The residue is maintained under high vacuum for 12 hours and the resulting white precipitate is recrystallized from hexane to give (3-2) as a white precipitate. ¹ H-NMR (CDCl 3): δ 7.38 (s, 1H), 5.03 (t, J = 8.2 Hz, 2H), 3.98 (s, 3H), 3.93 (s, 3H) ), 3.75 (t, J = 8.5 Hz, 2H).

The preparation of compound (3-3) is illustrated in Scheme 13. Methyl [4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -pyrazolo [1,5-a] pyrazin-2-yl] carboxylate - a compound of formula (3-3)

Under a nitrogen atmosphere, 14.0 g (48.0 mmol) of compound 3-2, 25 mL (144 mmol) of diisopropylethylamine, 12.0 g (52.6 mmol) of Boc-4- (amino- A solution of ethyl) piperidine and potassium iodide (2.39 g, 0.3 mmol) in acetonitrile (250 mL) was heated at reflux for 4.5 hours, cooled, filtered and concentrated under reduced pressure. The resulting yellow residue was purified by chromatography on silica gel using ethyl acetate as eluent. An off-white crystalline precipitate is obtained which gives 3-3.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.15 (s, 1H), 4.29 (t, J = 7.0 Hz, 2H), 3.93 (bd, J = 12 Hz, 2H), 3.76 (s, 3H), 3.61 (t, J = 5.3 Hz, 2H), 3.42 (t, J = 7.3 Hz, 2H), 2.65 (t, J = 7.6 Hz, 2H), 1.55 (d, J = 12.5 Hz, 2H), 1.38 (m, 2H), 1.33-1.25 (m, 1H), 1, 27 (s, 9H), 1.01 (m. 2H).

To a solution of (3-3) (90.81 mg, 0.223 mmol) in methanol (10 mL) was added a solution of lithium hydroxide (14.05 g, 0.335 mmol) in water (10 mL) and the resulting mixture was heated to 60 ° C. , For 5 hours. The reaction mixture was cooled and the methanol was removed under reduced pressure. The residual aqueous phase was acidified with 10% aqueous citric acid solution and extracted with dichloromethane (2 x 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate and evaporated to give the desired acid as a white precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.43 (s, 1H), 4.48 (t, J = 7.0 Hz, 2H), 4.01 (bd, J = 12 Hz, 2H), 3.77 (t, J = 5.3 Hz, 2H), 3.51 (t, J = 7.3 Hz, 2H), 2.71 (t, J = 8.3 Hz, 2H) , 1.72 (d, J = 12.5 Hz, 2H), 1.53 (m, 2H), 1.42-1.37 (m, 1H), 1.35 (s, 9H), 1, 10 (m, 2H).

Compound (3-4) can be prepared as shown in Scheme 14.

-40 (2S) - [(n-butylsulfonyl) amino-3 - / [{4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4- yl) ethyl] pyrazolo [1,5-a] pyrazin-2-yl) carbonyl] amino] -propionic acid - Compound 3-4

To a solution of 3.98 g (4.98 g, 12.72 mmol) in N-methylmorpholine (1.53 mL, 14.00 mmol) in THF (100 mL) was added 1.75 mL. (13.35 mmol) of isobutyl chloroformate and the resulting reaction mixture was stirred under a dry nitrogen atmosphere. After 1 hour, analysis of an aliquot by high performance liquid chromatography showed more than 90% complete reaction. The precipitated N-methylmorpholine hydrochloride is filtered off and the filtrate is poured into 4.30 g (16.54 mmol) of A-4, 4.27 mL (33.10 mmol) of diisopropylethylamine, 60 ml of tetrahydrofuran and 20 ml of water. The tetrahydrofuran is then removed under reduced pressure and the remaining aqueous portion is acidified with a saturated aqueous solution of potassium bisulfate. The mixture was extracted with ethyl acetate (3 x 200 mL), and the combined extracts were dried over sodium sulfate, filtered and evaporated. This gives a red oil from which the compound (3-4) precipitates as a white precipitate.

¹ H-NMR (DMSO-d 6): δ 8.31 (t, J = 6 Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.01 (s,

1H), 4.43 (t, J = 6.6 Hz, 2H), 4.11 (m, 1H), 3.92 (d, J = 12 Hz, 2H), 3.80 (t, J = 6.6 Hz, 2H), 3.51 (t, J = 7.3 Hz, 2H), 3.65 (m, 2H), 3.51 (t, J = 6.8 Hz, 2H), 2 , 96 (t, J = 7.2Hz, 2H), 1.70 (d, J = 11Hz, 2H), 1.53 (m, 2H), 1.60 -1.49 (overlapping m, 5H). ), 1.40 (s, 9H), 1.28 (q, J = 7.1 Hz, 2H), 1.05 (m, 2H), 0.79 (t, J = 7.1 Hz, 3H) ).

-41 2 (S) - [(n-Butylsulfonyl) amino] -3 - [[4,5,6,7-tetrahydro-4-oxo-5- [2- (piperidin-4-yl)] -ethyl] -pyrazolo [1,5-a] pyrazin-2-yl) carbonyl] -amino-propionic acid monohydrochloride - Compound 3-5

A solution of 3-4 (278 mg, 0.437 mmol) in ethyl acetate (30 mL) was cooled to 0 ° C and hydrogen chloride gas was bubbled through for 3 min. The reaction mixture was warmed to room temperature, stirred for 30 minutes and then evaporated to dryness in a rotary evaporator. The residual white precipitate was recrystallized from ethanol / water (90/10) and then filtered and dried in vacuo over phosphorus pentoxide. This gave compound (3-5) as a white precipitate. ¹ H-NMR (DMSO-d 6): δ 8.95 (bs, 1H), 8.33 (t, J = 5.7 Hz, 1H), 7.64 (d,

J = 9 Hz, 1H), 7.02 (s, 1H), 4.35 (t, J = 5.1 Hz, 2H), 4.10 (m, 1H), 3.81 (t,

J = 5.2 Hz, 2H), 3.6-3.4 (m, 4H), 3.21 (d, J = 10.5 Hz, 2H), 2.95 (t,

J = 7.8 Hz, 2H), 2.81 (br m, 2H), 1.96 (d, J = 11 Hz, 2H), 1.62-1.2 (overlapping multiplets, 9H), 0, 80 (t, J = 7.3 Hz, 2H).

The compound of formula 4-1a is prepared as shown in Scheme 15.

Methyl 2 (S) -N-Benzyloxycarbonylamino-3-aminopropionate hydrochloride - Compound 4-1a

The commercially available 2 (S) -N-benzyloxycarbonylamino-3-aminopropionic acid, available from Fluka, was refluxed in methanolic hydrochloric acid for 2.5 hours, then the reaction mixture was concentrated and crystallize from a mixture of methanol and diethyl ether. This gives a white precipitate of 4-1a.

-42 ^1H-NMR (300 MHz, DMSO-d _e): δ 7.63 (m, 5H), 5.93 (d, 1H), 5.15 (s, 2H), 4.56 (m, 1 H), 3.95-3.83 (m, 2H), 3.73 (s, 2H).

The compound of formula (4-3) can be prepared as shown in Scheme 16.

Methyl 2 (S) - [(CBZ) -amino] -3 - [[4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) - Ethyl] pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] amino] propionate - Compound 4-2

5.6 g (14.0 mmol) of compound (3-3) under a nitrogen atmosphere,

4.5 g (15.5 mmol) of methyl N- _a- Cbz-L-2,3-diaminopropionic acid hydrochloride,

A solution of HOBT (2.37 g, 15.5 mmol) and triethylamine (4.1 mL, 29.5 mmol) in dry dimethylformamide (65 mL) was stirred at room temperature for 48 hours. The dimethylformamide was removed under reduced pressure and the residue Dissolve in 700 ml of ethyl acetate. The resulting solution was washed successively with 100 ml of saturated aqueous sodium bicarbonate solution, 100 ml of water, 100 ml of 10% citric acid solution, 100 ml of water and 100 ml of a saturated aqueous solution of sodium chloride. dried over sulfate, filtered and evaporated. The resulting clear glassy precipitate was purified by chromatography on silica gel using dichloromethane (3% methanol) as eluent. The pure compound (4-2) is thus obtained in the form of a white precipitate. ¹ H-NMR (CDCl ₃ ): δ 7.43 (m, 5H), 7.35 (s, 1H), 7.18 (t, J = 6.5 Hz, 1H), 5.98 (d,

J = 6.8 Hz, 1H), 5.09 (s, 2H), 4.59 (m, 1H), 4.38 (m, 2H), 4.10 (br d, J = 12 Hz, 2H) ), 3.8 (s, 3H), 3.73 (t, J = 5.3 Hz, 2H), 2.71 (t, J = 8.3 Hz, 2H),

1.72 (d, J = 12.5 Hz, 2H), 1.53 (m, 2H), 1.42-1.37 (m, 1H), 1.35 (s, 9H),

1.10 (m, 2H).

Methyl 2 (S) -amino-3 - [[4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -pyrazolo [1 , 5-a] pyrazin-2-yl} carbonyl] amino] propionate - 4-3

To a solution of 4-2 (6.3 g, 10.26 mmol) in methanol (700 mL) was added 650 mg of 10% palladium on carbon and the resulting mixture was stirred under a hydrogen atmosphere at 1 atm for 48 hours. The catalyst was then removed by filtration through Celite and the filtrate was evaporated. The resulting colorless glass was triturated with diethyl ether and filtered. This gives a compound of formula (4-3) as a white precipitate. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.43 (m, 5H), 7.35 (s, 1H), 7.18 (t, J = 6.5 Hz, 1H),

5.98 (d, J = 6.8 Hz, 1H), 5.09 (s, 2H), 4.59 (m, 1H), 4.38 (m, 2H), 4.10 (broad d, J = 12 Hz, 2H), 3.81 (s, 3H), 3.73 (t, J = 5.3 Hz, 2H), 2.71 (t, J = 8.3 Hz, 2H), 1 , 72 (d, J = 12.5 Hz, 2H), 1.53 (m, 2H), 1.42-1.37 (m, 1H), 1.35 (s, 9H), 1.10 ( m, 2H).

Compound 4-4 is prepared as shown in Scheme 17.

Methyl 2 (S) - (acetylamino) -3 - {[{4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -ethyl] -pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] amino] -propionate - 4-4

To a solution of (4-3) (350 mg, 0.69 mmol) in tetrahydrofuran (10 mL) cooled to 0 ° C was added acetic anhydride (70 mL, 0.76 mmol) and the reaction mixture was allowed to warm to room temperature. After stirring for 18 hours, the reaction mixture was concentrated. The residue was dissolved in ethyl acetate (50 mL) and the resulting solution was stirred

-44 was then washed with 25 ml of sodium bicarbonate solution, 25 ml of water, 25 ml of 10% potassium hydrogen sulphate solution, 25 ml of water and 25 ml of a saturated aqueous sodium chloride solution, and the organic phase was dried over anhydrous. dried over sodium sulfate and evaporated. The resulting colorless residue was chromatographed on silica gel, eluting with 3% methanol in dichloromethane, to give compound 4-4 as a white precipitate.

¹ H-NMR (CDCl ₃ ): δ 7.29 (s, 1H), 7.24 (t, J = 6.4 Hz, 1H), 6.81 (d, J = 7.6 Hz, 1H). .

4.79 (m, 1H), 4.38 (m, 2H), 4.10 (br d, J = 12 Hz, 2H), 3.81 (s, 3H),

3.80 (m, 2H), 3.73 (t, J = 5.3 Hz, 2H), 2.71 (t, J = 8.3 Hz, 2H), 2.01 (s, 3H),

1.72 (d, J = 12.5 Hz, 2H), 1.57 (m, 2H), 1.42-1.37 (m, 1H), 1.37 (s, 9H),

1.09 (m, 2H).

2 (S) - (Acetylamino) -3 - / [{4,5,6,7-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) ethyl] pyrazolo [1, 5-a] Pyrazin-2-yl} carbonyl] -amino-propionic acid - 4-5

A solution of 4-3 (203 mg, 0.38 mmol) in 1 N lithium hydroxide (0.76 mL, 0.76 mmol) in water (5 mL), methanol (5 mL) and tetrahydrofuran (5 mL) at room temperature overnight the organic solvents are evaporated under reduced pressure. The resulting solution was diluted with water (25 mL), acidified with 10% potassium bisulfate solution and then extracted with ethyl acetate. The extract was washed with water and then with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and evaporated to give the desired acid.

¹ H-NMR (CDCl 3): δ 7.93 (br, 1H), 7.81 (br, 1H), 7.29 (s, 1H), 4.79 (m,

1 H), 4.348 (m, 2H), 4.10 (br d, J = 12 Hz, 2H), 3.80 (br m, 2H),

3.73 (broad, t, 2H), 2.71 (t, J = 8.3 Hz, 2H), 2.11 (s, 3H), 1.72 (d,

-45J = 12.5 Hz, 2H), 1.57 (m, 2H), 1.42-1.37 (m, 1H), 1.37 (s, 9H), 1.12 (m, 2H) .

169 mg (32.8 mmol) of this acid are dissolved in 50 ml of ethyl acetate and the resulting solution is cooled to 0 ° C and treated with dry gaseous hydrogen chloride for 30 minutes. The solution was concentrated in vacuo and the residue was triturated with anhydrous diethyl ether, filtered and dried over phosphorus pentoxide. This gave compound 4-5 as a white solid, m.p. 150-156 ° C.

The conversion of 4-4 into 4-5 is shown in Scheme 18.

The preparation of compound (4-7) is illustrated in Scheme 19. 2 (S) - [(Cbz-Amino)] - 3 - / [{[2- (N-Cbz-piperidin-4-yl) -4,5,6,7-tetrahydro-4-oxo-5- ethyl ] -pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] amino] -propionsa v- (4-6)

If, as described for the preparation of (3-4) compound, the compound (4-3) can be obtained from Fluka (4-1) is coupled with N _to N -Cbz-L-2,3-diaminopropionic acid, the (4 The compound of formula -6, i.e. the doubly protected adduct, is obtained.

2 (S) -Amino-3 - / [{4,5,6,7-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) ethyl] pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] -amino-propionic acid - Compound 4-7

Treatment of the compound (4-6) with hydrogen gas in the presence of palladium on carbon gives the desired acid having a melting point of 157 ° C. The Boc group is then removed in a conventional manner.

-46-chloride in ethyl acetate to give the pure compound 4-7, m.p. 195-198 'C.

The compound of formula (4-8) can be prepared as shown in Scheme 20.

Methyl 2 (S) - [(n-butylsulfonylamino)] - 3 - [[4,5,6,7-tetrahydro-4-oxo-5- [2- (N-CBZ-piperidine-4) -yl) ethyl] pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] amino] propionate (4-8)

A solution of 4- (3) (0.30 g, 0.61 mmol), n-butylsulfonyl chloride (0.16 g, 0.91 mmol) and tetrahydrofuran (50 mL) was added at room temperature for 12 hours. the solvent is evaporated under reduced pressure. The resulting oil was dissolved in dichloromethane (50 mL), and the resulting solution was washed with 10% potassium bisulfite (50 mL), dried over anhydrous sodium sulfate, filtered and evaporated. The resulting residue was chromatographed on silica gel to give 4-8 as a colorless glass.

¹ H-NMR (CDCl 3): δ 8.31 (t, J = 6 Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.01 (s, 1H),

4.43 (t, J = 6.6 Hz, 2H), 4.11 (m, 1H), 3.92 (d, J = 12 Hz, 2H), 3.83 (s, 3H),

3.80 (t, J = 6.6 Hz, 2H), 3.51 (t, J = 7.3 Hz, 2H), 3.65 (m, 2H), 3.51 (t, J = 6) , 8 Hz, 2H), 2.96 (t, J = 7.2 Hz, 2H), 1.70 (d, J = 11 Hz, 2H), 1.53 (m, 2H), 1.60 - 1.49 (overlapping m, 5H), 1.40 (s, 9H), 1.28 (q, J = 7.1 Hz, 2H), 1.05 (m, 2H), 0.79 (t, J = 7.1 Hz, 3H).

The compound (5-8) is prepared according to the sequence outlined in Scheme 21.

«·

-47Dimethyl 1- (3-bromopropyl) pyrazole-3,5-dicarboxylate - Compound 5-1

As described for the preparation of compound (3-2), compound (5-1) can be obtained from compound (3-1a) using 1,3-dibromopropane as a white crystalline precipitate.

¹ H-NMR (CDCl ₃ ): δ 7.38 (s, 1H), 4.95 (t, J = 8.2 Hz, 2H), 3.95 (s, 3H), 3.92 (s,

3H), 3.75 (t, J = 8.5 Hz, 2H), 2.51 (m, 2H).

Dimethyl 1- (3-azidopropyl) pyrazole-3,5-dicarboxylate - (5-2)

To a solution of (5-1) (1.0 g, 3.45 mmol) in dimethyl sulfoxide (10 mL) was added sodium azide (0.883 g, 13.8 mmol), and the resulting mixture was stirred at 25 ° C for 5 hours. stirred. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined extracts were washed with water (2 x 100 mL) followed by brine (100 mL), dried over anhydrous sodium sulfate and evaporated to give 5-2 as a colorless oil.

Methyl [5,6,7,8-tetrahydro-4-oxo-4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl] carboxylate (5-3)

To a solution of compound (5-2) (851 mg, 3.25 mmol) in dry ethanol (100 mL) was added palladium on carbon (100 mg, 10% by weight) and the resulting mixture was shaken in a Parr hydrogenator at 45 pounds per square inch for 5 hours. . The catalyst was filtered off with celite and the filtrate was evaporated to give 800 mg of a colorless oil. NMR analysis of the oil showed that 1- (3-aminopropyl) dimethylpyrazole-3,5-dicarboxylate »· 4 4 · <* · <

·· · · * · ·· ··· * «

-48and a mixture of cyclic diazepinone. This mixture was dissolved in benzene (50 mL) and the resulting solution was refluxed for 15 hours and then concentrated. The tan precipitate thus obtained was recrystallized from a mixture of dichloromethane and hexane to give 5-3 as a white solid, m.p. 220-221 ° C.

¹ H-NMR (CDCl 3): δ 7.36 (s, 1H), 6.42 (broad t, 1H), 4.58 (t, J = 8.0 Hz, 2H), 3.95 (s, 3H), 3.39 (dt, J = 7.2 Hz, 2H), 2.31 (m, 2H).

Methyl [5,6,7,8-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -4H-pyrazolo [1,5-a] [1,4] ] diazepin-2-yl] carboxylate - (5-5)

To a solution of 5-3 (175 mg, 0.83 mmol) in dimethylformamide (50 mL) was added sodium hydride (60 mg, 60 mg, 0.91 mmol) and the resulting mixture was stirred at -15 ° C under nitrogen. Stir at C for 30 minutes. A solution of 2- (N-Boc-piperidin-4-yl) -ethyl iodide (283 mg, 0.83 mmol) in dimethylformamide (25 mL) was added dropwise over 20 minutes. The resulting reaction mixture was stirred at -15 ° C for 30 minutes and then allowed to warm to room temperature. After stirring overnight, the dimethylformamide was evaporated under reduced pressure and the residue redissolved in ethyl acetate. The resulting solution was filtered and purified by chromatography on silica gel using ethyl acetate as the eluent. This gives the pure compound (5-5) as a glassy material.

¹ H-NMR (CDCl 3): δ 7.24 (s, 1H), 4.50 (t, J = 7.0 Hz, 2H), 3.93 (broad d,

J = 12 Hz, 2H), 3.94 (s, 3H), 3.61 (t, J = 5.3 Hz, 2H), 3.42 (t, J = 7.3 Hz, 2H), 2 , 7 (broad t, J = 6.3 Hz, 2H), 2.3 (m, 2H), 1.55 (d, J = 12.5 Hz, 2H),

1.38 (m, 2H), 1.33-1.25 (m, 1H), 1.27 (s, 9H), 1.01 (m, 2H).

• •• 9 «

-49 [5,6,7,8-tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) ethyl-4H-pyrazolo [1,5-a] [1,4 ] diazepin-2-yl] carboxylic acid - Compound 5-6

To a solution of 5-5 (166 mg, 0.395 mmol) in methanol (10 mL) was added 1N sodium hydroxide (0.435 mL, 0.43 mmol), and the resulting mixture was stirred at room temperature for 18 hours and then methanol was reduced. removed under pressure. The residual aqueous phase was acidified with 10% aqueous citric acid solution and extracted with dichloromethane (2 x 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate and evaporated to give (5-6) as a white precipitate.

¹ H-NMR (CDCl 3): δ 7.29 (s, 1H), 4.52 (t, J = 7.0 Hz, 2H), 4.12 (broad d,

J = 12 Hz, 2H), 3.94 (s, 3H), 3.61 (t, J = 5.3 Hz, 2H), 3.42 (t, J = 7.3 Hz, 2H), 2 , 7 (broad t, J = 6.3 Hz, 2H), 2.3 (m, 2H), 1.55 (d, J = 12.5 Hz, 2H),

1.38 (m, 2H), 1.33-1.25 (m, 1H), 1.27 (s, 9H), 1.01 (m, 2H).

Ethyl 2 (S) - [(n-butylsulfonyl) amino] -3 - [[5,6,7,8-tetrahydro-4-oxo-5- [2- (N-Boc-piperidine-4) -yl) ethyl] -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl} carbonyl] aminoZ-propionate (5-7)

To a solution of 5-6 (147 mg, 0.36 mmol) in dichloromethane (5 mL) was added ethyl 2 (S) -n-butanesulfonamido-3 (115 mg, 0.40 mmol). aminopropionate, HOBT (49 mg, 0.36 mmol) and triethylamine (0.10 mL, 0.724 mmol) in dichloromethane (50 mL). The reaction mixture was stirred under nitrogen at room temperature for 18 hours, then successively with 20 ml of saturated sodium bicarbonate solution, 20 ml of water, 20 ml of 10% citric acid solution, 20 ml of water and 20 ml of saturated aqueous sodium chloride solution. Over anhydrous sodium sulfate

After drying, filtration and evaporation, the resulting clear glass material was purified by chromatography on silica gel using 5% methanol in ethyl acetate as eluent. This gives the pure compound (5-7).

¹ H-NMR (CDCl 3): δ 7.32 (s, 1H), 7.24 (t, J = 6.8 Hz, 1H), 5.54 (t, J = 7.2 Hz, 1H),

4.43 (t, J = 7.8 Hz, 2H), 4.32 (m, 1H), 4.28 (q, J = 7.1 Hz, 2H), 4.10 (broad d, J = 12 Hz, 2H), 3.85 (m, 2H), 3.61 (t, J = 5.3 Hz, 2H), 3.42 (t, J = 7.3 Hz, 2H), 3.03 (t, J = 7.1 Hz, 2H), 2.7 (broad t, J = 6.3 Hz, 2H), 2.3 (m, 2H), 1.65 -1.45 (overlapping m, 7H), 1.38 (m, 2H), 1.33-1.25 (m, 1H),

1.37 (s, 9H), 1.30 (t, J = 7.4 Hz, 3H), 1.01 (m, 2H), 0.96 (t, J = 7.3 Hz, 3H). 2 (S) - [(n-butylsulfonyl) amino] -3 - / [{5,6,7,8-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) ethyl ] -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl} carbonyl] -amino} -propionic acid - Compound 5-8

To a solution of 5-7 (100 mg, 0.156 mmol) in methanol (10 mL) was added 1N sodium hydroxide (160 mL, 0.16 mmol) and water (10 mL), and the resulting solution was stirred at room temperature for 3.5 hours. stirred. The methanol is then removed under reduced pressure and the remaining aqueous phase is acidified with 10% aqueous citric acid. The mixture was extracted twice with dichloromethane (50 ml) and the combined organic extracts were dried over anhydrous sodium sulfate and concentrated to give the desired acid.

¹ H-NMR (CDCl 3): δ 17.24 (t, J = 6.8 Hz, 1H), 7.28 (s, 1H), 6.0 (d, J = 7.2 Hz, 1H),

4.43 (t, J = 7.8 Hz, 2H), 4.32 (m, 1H), 4.10 (broad d, J = 12 Hz, 2H), 3.85 (m, 2H), 3 , 61 (t, J = 5.3 Hz, 2H), 3.42 (t, J = 7.3 Hz, 2H), 3.03 (t, J = 7.1 Hz, 2H), 2.7 (broad t, J = 6.3 Hz, 2H), 2.3 (m, 2H), 1.65-1.45 (overlapping m,

7Η), 1.38 (m, 2H), 1.33 - 1.25 (m, 1H), 1.37 (s, 9H), 1.01 (m, 2H), 0.96 (t, J = 7.3 Hz, 3H).

A solution of this acid (89 mg) in ethyl acetate (15 mL) was cooled to 0 ° C and hydrogen chloride gas was bubbled through it for 3 min. The reaction mixture was then warmed to room temperature, stirred for 30 minutes and finally evaporated to dryness in a rotary evaporator. The residual white precipitate was triturated with diethyl ether, filtered and dried in vacuo over phosphorous pentoxide to give 5-8 as a white precipitate. ¹ H-NMR (DMSO-d 6): δ 8.59 (bs, 1H), 8.33 (t, J = 5.7 Hz, 1H), 7.64 (d,

J = 9 Hz, 1H), 7.02 (s, 1H), 4.35 (t, J = 5.1 Hz, 2H), 4.10 (m, 1H), 3.81 (t,

J = 5.2 Hz, 2H), 3.6-3.4 (m, 4H), 3.21 (d, J = 10.5 Hz, 2H), 2.95 (t,

J = 7.8 Hz, 2H), 2.81 (br m, 2H), 1.96 (d, J = 11 Hz, 2H), 1.62-1.2 (overlapping multiplets, 9H), 0, 80 (t, J = 7.3 Hz, 2H).

The compound of formula (6-3) can be prepared as shown in Scheme 22.

Ethyl [3-oxo-2- [2- (N-Boc-piperidin-4-yl) -ethyl] -octahydro-imidazo [1,5-a] pyridin-6-yl] -carboxylate (6-1A) and compounds 6-1B

To a solution of compound 2-6 (514 mg, 1.1 mmol) in acetone (25 mL) was added water (10 mL) and the mixture was heated to 60 ° C.

3.5 hours. The acetone is then removed under reduced pressure and the yellow precipitate formed is filtered off. This crude material was dissolved in 100 mL of toluene and refluxed for 3 hours. The toluene was then evaporated to give a yellow precipitate.

500 mg (1.11 mmol) of this precipitate is dissolved in 100 ml of ethanol and 75 mg of 10% w / w of carbon are added to the resulting solution.

-52 Palladium catalyst is added. The reaction mixture was shaken for 13 hours in a Parr hydrogenator at 55 pounds per square inch. The catalyst is then filtered off using Celite filtration aid and the solvent is evaporated from the filtrate. The resulting colorless oil was purified by chromatography on silica gel using a mixture of 70% ethyl acetate and 30% hexane as eluent. 268 mg of cis product 6-1A and 132 mg of trans product 6-1B are obtained.

(6-1 A) isomer:

¹ H-NMR (CDCl ₃ ): δ 4.15 (m, 1H), 4.06 (m, 2H), 3.68 (s, 3H), 3.41 (m, 2H), 3.22 ( m, 1H), 2.90 (m, 1H), 2.80 (m, 2H), 2.68 (m, 2H), 2.43 (m, 1H), 2.16 (m, 1H), 1.81 (m, 1H), 1.69 (m, 2H), 1.60 (m, 1H), 1.45 (s, 9H), 1.43 (m, 3H),

1.39 (m, 1H), 1.11 (m, 2H).

(6-1B) isomer:

¹ H-NMR (CDCl 3): δ 4.34 (m, 1H), 4.06 (m, 2H), 3.69 (s, 3H), 3.40 (m, 3H), 3.36 (m , 1H), 3.30 (m, 1H), 3.11 (m, 1H), 2.89 (m, 1H), 2.84 (m, 1H), 2.67 (m, 2H), 2 , 63 (m, 1H), 2.30 (m, 1H), 1.69 (m, 2H), 1.67 (m, 2H), 1.60 (m, 1H), 1.45 (s, 9H), 1.41 (m, 1H), 1.09 (m, 2H).

Tert-Butyl (±) -cis - [[3-oxo-2- [2- (N-Boc-piperidin-4-yl) -ethyl] -octahydro-imidazo [1,5-a] pyridine-6- (ii) carbonyl] amino propionate (6-2)

In a mixture of methanol and water, compound (6-1A) is hydrolyzed with 1N sodium hydroxide as described for the preparation of compound (1-6) to give the desired acid. This is then coupled with β-alanine tert-butyl ester as described for the preparation of compound (2-8) to give compound (6-2).

¹ H-NMR (CDCl ₃ ): δ 6.32 (t, 1H), 4.06 (m, 2H), 3.98 (m, 1H), 3.56 (m, 5H), 3.21 ( m, 2H), 2.91 (m, 1H), 2.92 (m, 1H), 2.80 (m, 2H), 2.40 (t, 2H), 2.23 (m, 1H), 2.09 (m, 1H), 1.81 (m, 3H), 1.69 (m, 2H), 1.60 (m, 1H), 1.45 (s, 18H), 1.43 (m , 3H), 1.11 (m, 2H).

(±) cis - / [{3-Oxo-2- [2- (pipendin-4-yl) ethyl] octahydro-imidazo [1,5-a] pyridin-6-yl} carbonyl] amino Propionic acid - a compound of formula (6-3)

A solution of 6-2 (65.3 mg, 0.13 mmol) in dichloromethane (10 mL) was cooled to 0 ° C and trifluoroacetic acid (0.200 mL) was added. The resulting solution is stirred for 1 hour and then evaporated to dryness under reduced pressure to give pure 6-3.

¹ H-NMR (CD 3 OD): δ 4.06 (m, 2H), 3.91 (m, 1H), 3.56 (m, 5H), 3.21 (m, 2H),

2.91 (m, 1H), 2.92 (m, 1H), 2.80 (m, 2H), 2.40 (t, 2H), 2.23 (m, 1H), 2.09 (m) , 1H), 1.81 (m, 3H), 1.68 (m, 2H), 1.60 (m, 1H), 1.28 (m, 3H), 0.91 (m, 2H).

The compound of formula (7-3) can be prepared as shown in Scheme 23.

Methyl (2S) - [(N-CBZ-aminosulfonyl) amino] -3 - [[4,5,6,7-tetrahydro-4-oxo-5- [2- (N-BOC-piperidine)] 4-yl) ethyl] pyrazolo [1,5-a] pyrazin-2-yl} carbonyl] aminoZ-propionate - Compound 7-1

To a solution of chlorosulfamyl isocyanate (45.1 µL, 0.508 mmol) in methylene chloride cooled to 0 ° C was added benzyl alcohol (53 mL, 0.508 mmol) and the resulting mixture was allowed to stand at 0 ° C for 90 min. forget it. A solution of compound 4-3 (250 mg, 0.508 mmol) in methylene chloride (142 mL, 1.02 mmol) was then added. The reaction mixture was allowed to warm to room temperature and stirred for 18 hours. The reaction mixture was then adjusted to pH 3.0 with aqueous sodium hydrogen sulfate solution and extracted with methylene chloride (3 x 10 mL). The organic extracts were combined, evaporated and the residue was purified by chromatography on silica gel using a 95% (v / v) mixture of methylene chloride and 5% (methanol) as eluent. The compound of formula (7-1) is obtained. ¹ H-NMR (300 MHz, CDCl ₃ ): δ 1.40 (s, 9H), 2.65 (t, 2H), 3.65 (s, 3H), 5.10 (s, 2H), δ , 65 (d, 1H), 7.2-7.5 (m, 6H), 8.90 (s, 1H).

2 (S) - [(N-CBZ-Aminosulfonyl) amino] -3 - / [{4,5,6,7-tetrahydro-4-oxo-5- [2- (N-Boc-piperidine 4-yl-ethyl] -pyrazolo [1,5-a] pyrazin-2-yl} -carboxyl] -amino} -propionic acid - Compound 7-2

To a solution of compound (7-1) (100 mg) in tetrahydrofuran (5 mL) was added lithium hydroxide solution (0.6 mL) and the resulting mixture was stirred at room temperature for 18 hours. The reaction mixture is quenched by the addition of aqueous sodium hydrogen sulfate solution to pH 3.0 and extracted twice with 15 ml of ethyl acetate. Concentration of the extracts yields the desired acid.

¹ H-NMR: δ 1.4 (s, 9H), 2.6 (broad, t, 2H), 7.1-7.2 (broad m, 5H), 7.3 (s, 1H).

This acid was dissolved in ethyl acetate and the resulting solution was cooled to -5 ° C and then treated with gaseous hydrogen chloride. Finally, the reaction mixture was concentrated and rinsed with ethyl acetate. There is thus obtained a compound of formula (7-2) having a melting point higher than 200 ° C.

-55Analysis Results:

Calculated: C, 44.86; H, 5.92; N, 14.20;

Found: C, 44.50; H, 6.09; N, 13.80.

2 (S) - (Aminosulfonyl) amino-3 - / [{4,5,6,7-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) ethyl] pyrazolo [1 , 5-a] Pyrazin-2-yl) carbonyl] amino / propionic acid - To a solution of 7-2 mg of the compound 7-2 in 20 ml of methanol is added 35 mg of 10% palladium on carbon and then the reaction mixture was hydrogenated at 1 atm for 18 hours. The reaction mixture was filtered and the filtrate was evaporated to give 46 mg of an oil. This oil was dissolved in ethyl acetate and the solution cooled to 0 ° C and bubbled with hydrogen chloride gas for 30 minutes. Evaporation of the reaction mixture gave a white precipitate (7-3) having a melting point higher than 200 ° C. FAB mass spectrum, M + 1 = 458.

Compound 8-5 can be prepared as shown in Scheme 24.

Diethyl 1- (2-Bromoethyl) pyrrole-2,4-dicarboxylate (8-2)

A solution of diethyl pyrrole-2,4-dicarboxylate (5.50 g, 29.4 mmol) in tetrahydrofuran (200 mL) was cooled in an ice bath and added continuously.

A suspension of 6.5 g (68.6 mmol) of 60% sodium hydride in 50 ml of tetrahydrofuran was added. The reaction vessel was then warmed to room temperature and, after stirring for 1 hour at room temperature, 1,2-dibromoethane (25.2 mL, 294 mmol) was added. The reaction mixture was refluxed for 24 hours and water (50 mL) was added. Then the reaction

The reaction mixture was subjected to rotary evaporation in a rotary evaporator, and the residue was treated with 100 ml of saturated sodium bicarbonate solution, extracted 4 times with 50 ml of methylene chloride, and the combined organic extracts were dried over anhydrous sodium hydroxide. the desiccant is filtered off and the filtrate is evaporated in a rotary evaporator. The resulting yellow precipitate was recrystallized from hexane / ethyl acetate (80:20 by volume) to give 8-2 as a yellow precipitate.

¹ H-NMR (DMSO-d 6): δ 7.83 (d, 1H), 7.15 (d, 1H), 4.69 (t, 2H), 4.25 (m, 4H),

3.78 (t, 2H), 3.31 (H ₂ O), 1.29 to 1.22 (m, 6H).

Ethyl [4,5,6,7] -tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -pyrrolo [1,5-a] pyrazin-2-yl ] carboxylate 8-3

3.30 g (10.4 mmol, 1.0 equivalents) of alkyl bromide (8-2), 3.52 g (15.5 mmol, 1.5 equivalents) of compound (2-4), 5, A suspension of 18 g (31.2 mmol) of potassium iodide and 5.42 mL (31.2 mmol, 3.0 equivalents) of diisopropylethylamine in 50 mL of acetonitrile was refluxed for 24 hours and then the acetonitrile was rotated in a vacuum evaporator. removed. Saturated sodium bicarbonate solution (100 mL) was added to the residue, followed by extraction with ethyl acetate (5 x 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate and evaporated to give a brown oil. The latter is then purified by column chromatography on silica gel using methylene chloride as the eluant, followed by methylene chloride containing 1% (v / v) methanol. This afforded pure 8-3 as a white precipitate.

-57 ¹ H-NMR (CDCl): δ 7.32 (d, 1H), 7.29 (d, 1H), 4.28 (q, 2H), 4.19 to 4.00 (m, 4H) .

3.7 - 3.6 (t, 2H), 3.6 - 3.5 (t, 2H), 2.68 (t, 2H), 1.8 - 1.7 (m, broad, 2H, H) ₂ O), 1.6-1.4 (s, m, 11H), 1.33 (t, 3H), 1.2 to 1.1 (m, 2H).

Tert-Butyl [4,5,6,7] -tetrahydro-4-oxo-5- [2- (N-Boc-piperidin-4-yl) -ethyl] -pyrrolo [1,5-a] pyrazine-2 -yl] aminopropionate (8-4)

Into a 50 mL round bottom flask equipped with a magnetic stir bar was added 620 mg (1.54 mmol) of compound 8-3, 160 mg (4.00 mmol, 2.6 equiv.) Of lithium hydroxide monohydrate, 15 mL of water and 10 mL. methanol. The resulting solution was stirred at room temperature for 4 hours and then heated at 90 ° C for 1 hour. The residual methanol was then removed by rotary evaporation, the aqueous residue was acidified with 10% potassium sulfate solution and extracted with ethyl acetate (4 x 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and evaporated to give the desired acid as a white precipitate.

¹ H-NMR (DMSO-d 6): δ 7.51 (d, 2H), 6.84 (d, 1H), 4.19 (m, 2H), 3.90 (d, broad,

2H), 3.63 (m, 2H), 3.43 (m, 2H), 3.4-3.2 (H2O), 2.6-2.8 (broad, 2H),

1.66 (d, 2H), 1.43 (m, 2H), 1.36 (s, 9H), 1.1-0.9 (m, 2H).

Into a 100 ml round-bottomed flask equipped with a magnetic stirrer, 150 mg (554 mmol), 117 mg (0.609 mmol) EDC, 82.2 mg (0.609 mmol) 1-hydroxybenzothiazole, 0.300 ml (1, 11 mmol (4.0 equivalents) of triethylamine, 111 mg (0.609 mmol) of β-alanine tert-butyl ester and 10 ml of methylene chloride. The resulting solution was stirred at room temperature overnight and the solvent was removed in a rotary evaporator. The resulting residue was purified by column chromatography on silica gel using methylene chloride containing 0, 2 and 4% by volume of methanol as the eluent after one to 58 min. The product-containing fractions were combined and evaporated to give (8-4) as a white precipitate.

¹ H-NMR (CDCl ₃ ): δ 7.32 (d, 1H), 7.04 (d, 1H), 6.58 (m, 1H), 4.2 - 4.0 (m, 4H),

3.7 to 3.5 (m, 6H), 2.68 (t, 2H), 2.51 (t, 2H), 1.70 (m, 3H, H ₂ O), 1.53 (m, 2H), 1.45 (s, 9H), 1.21-1.0 (m, 2H).

4,5,6,7 - / [{tetrahydro-4-oxo-5- [2- (piperidin-4-yl) ethyl] [1,5-a] pyrazin-2-yl} carbonyl] amino N-Propionic acid (8-5) In a round-bottomed flask (180 ml, 0.347 mmol) was added compound (8-4) and ethyl acetate (10 ml), and the resulting slurry was cooled in an ice bath. Hydrogen chloride gas was bubbled through the suspension for 1.5 minutes, the reaction vessel was warmed to room temperature and the solvent was removed by vacuum filtration. 248 DEG-249 DEG C. as a white precipitate to give compound 8-5.

¹ H-NMR (DMSO-d ₆ ): δ 9.0 - 8.5 (broad, 2H), 8.05 (m, 1H), 7.42 (d, 1H), 7.05 (d, 1H) ), 4.15 (m, 2H), 3.62 (m, 2H), 3.5 - 3.3 (m, 4H, H ₂ O), 3.25 - 3.15 (d, broad, 2H) ), 2.85-2.7 (broad, 2H), 2.5-2.4 (m, 4H), 1.81 (d, 2H),

1.6-1.4 (m, 3H), 1.4-1.2 (m, 2H).

Compound 9-5 can be prepared as shown in Scheme 25.

Ethyl 2 (S) -amino-3- (N-Boc-amino) -propionate (9-1) g (96.2 mmol), 2 (S) -3-diaminopropanoic acid available from Fluka of anhydrous ethanol solution was saturated with anhydrous gaseous hydrogen chloride and refluxed for 2.5 hours. The solvent was removed and the residue was ethanol and diethyl ether

It is recrystallized from a mixture of -59 to give the corresponding ethyl ester dihydrochloride as a hygroscopic white precipitate.

5 g (24.3 mmol) of this material are suspended in 200 ml of dichloromethane cooled to -50 [deg.] C., then 7.0 ml (51 mmol) of triethylamine are added and the mixture is stirred for 5 minutes. . A solution of di-tert-butyl dicarbonate (5.30 g, 24.3 mmol) in dichloromethane (100 mL) was added dropwise over 30 min, and the reaction mixture was stirred at -50 ° C for 1.5 h and then heat to room temperature. The mixture is washed twice with 100 ml of water each and then dried over anhydrous sodium sulfate and evaporated. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane: methanol = 80:20, v / v). This gives pure 9-1.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 6.03 (broad t, 1H), 4.35 (m, 1H), 3.85 (m, 2H),

3.23 (m, 2H), 1.21 (t, 3H).

Ethyl 2 (S) - (n-butylaminosulfonylamino) -3- (N-BOC-amino) propionate - Compound 9-2

To a solution of 9-1 (500 mg, 2.15 mmol) in dichloromethane (10 mL) was added pyridine (261 mL, 3.23 mmol) and n-butylsulfamoyl chloride (406 mg, 2.37 mmol). the resulting solution was stirred at room temperature for 3 hours and then poured onto silica gel. Elution with 30% acetone in hexane afforded pure 9-2 as a white precipitate.

-60 ^1H-NMR (300 MHz, _CDCl3): δ 5.43 (d, 1H), 4.98 (t, 1H), 4.40 (br s, 1H);

4.10 (q, 2H), 4.05 (m, 1H), 3.56 (m, 2H), 3.08 (m, 2H), 1.8-1.2 (overlapping multiplets, 16H), 0.90 (t, 3H).

Ethyl 2 (S) - (n-butylaminosulfonylamino) -3-aminopropionate - Compound 9-3

To a solution of 9-2 (612 mg, 1.65 mmol) in ethyl acetate (50 mL) at -5 ° C was bubbled anhydrous hydrogen chloride at -5 ° C for 30 minutes, then the reaction mixture was concentrated and the product was isolated by filtration. to give 9-3 as a white precipitate.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 5.82 (d, 1H), 4.56 (bs, 1H), 4.20 (q,

2H), 4.02 (m, 1H), 3.45 (m, 2H), 3.01 (m, 2H), 1.9-1.36 (m, 7H), 0.93 (t, 3H). ).

Ethyl 2 (S) - [(n-butylaminosulfonyl) amino] -3- [4,5,6,7-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) -ethyl] -pyrazolo [1,5-a] pyrazin-2-yl] carbonyl-aminopropionate - (9-4)

The compound (9-3) is coupled with the compound (3-3) in dimethylformamide by EDC and HOBT as described for the preparation of the compound (1-6) to give the compound (9-4).

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.29 (s, 1H), 7.19 (t, 1H), 5.43 (d, 1H), 4.41 (t,

2H), 4.26 (q, 2H), 4.20 (m, 1H), 4.08 (d, 2H), 3.86 (m, 2H), 3.76 (m, 2H),

3.60 (t, 2H), 3.08 (t, 2H), 2.68 (t, 2H), 1.78 (d, 2H), 1.6 - 1.08 (m, 8H),

1.43 (s, 9H), 0.92 (t, 3H).

-61 2 (S) - [(n-Butylaminosulfonyl) amino] -3 - [[4,5,6,7-tetrahydro-4-oxo-5- (piperidin-4-yl) - ethyl] -pyrazolo [1,5-a] pyrazin-2-yl) carbonyl] amino] -propionic acid - Compound 9-5

As described for the preparation of compound 5-7, the compound 9-4 is hydrolyzed with sodium methylate and the crude acid is isolated and then treated with hydrogen chloride in ethyl acetate. 155-160 ° C as a white precipitate to give compound 9-5.

Compound (10-4) can be prepared as shown in Scheme 26.

N- [2- (5-Methoxycarbonyl) pyridylmethyl] -? - alanine tert-butyl ester - Compound 10-1

Into a 250 ml flask was placed 871 mg (3.79 mmol) of compound (2-3), 2.7 g (15 mmol) of β-alanine tert-butyl ester hydrochloride and 4.5 g (30 mmol) of potassium. carbonate in 100 ml of anhydrous acetonitrile and refluxed for 3 hours. The reaction mixture was cooled and filtered. The filtrate was concentrated under reduced pressure and the residue was chromatographed on silica gel, eluting with ethyl acetate. This gives compound 10-1 as a colorless glass.

¹ H-NMR (CDCl ₃ ): δ 9.18 (d, J = 1.4 Hz, 1H), 8.1 (dd, J = 1.4 and 6.8 Hz, 1H), 7.39 ( d, J = 6.8 Hz, 1H), 4.08 (s, 2H), 3.95 (s, 3H), 3.04 (t, 2H), 2.60 (t, 2H),

1.4 (s, 9H).

· · ·

-62-tert-Butyl 2- (2-carboxyethyl) -1- (chlorocarbonyl) -3-oxo-2,3-dihydroimidazo [1,5-a] pyridine-6-carboxylate - (10- 2)

To a solution of 10-1 (800 mg, 2.71 mmol) in toluene (50 mL) cooled to 0 ° C was added NN-dimethylaniline (2.0 mL, 16.7 mmol), followed by 30 min. solution of triphosgene (1.7 g, 5.7 mmol) in toluene (15 mL) was added dropwise. The reaction mixture was heated to 25 ° C and stirred for 3 hours. It is then washed twice with 50 ml of 1 N hydrochloric acid, 50 ml of water and 50 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give compound 10-2 as a yellow crystalline solid.

¹ H-NMR (CDCl ₃ ): δ 8.83 (d, J = 1.4 Hz, 1H), 8.25 (d, J = 6.8 Hz, 1H), 7.82 (dd, J = 1.4 and 6.8 Hz, 1H), 4.43 (t, J = 7.2 Hz, 2H), 3.98 (s, 3H), 2.75 (t, J = 7.2 Hz, 2H), 1.4 (s, 9H).

Tert-Butyl 2- (2-carboxyethyl) -1-cyano-3-oxo-2,3-dihydroimidazo [1,5-a] pyridine-6-carboxylate - Compound (10-3)

To a solution of compound (10-2) (250 mg, 0.65 mmol) in dichloromethane (100 mL) was added a solution of ammonium hydroxide (10 mL) and the resulting biphasic mixture was stirred for 1 hour. The organic layer was separated, washed with 10% citric acid solution, then with 50 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated.

This residue (150 mg, 0.42 mmol) was dissolved in dichloromethane (100 mL), and methoxycarbonylsulfamoyl triethylammonium hydroxide inner salt (Burgess- 355.7 mg, 1.48 mmol) was added to the resulting solution. reagent) in three doses over a 2 hour period. The resulting solution was stirred at room temperature for an additional 1 hour and then concentrated. The residue was purified on silica gel

Purify by -63 chromatography eluting with hexane / ethyl acetate (1: 1). This gives the desired nitrile compound in quantitative yield. This compound is then saponified with 1 N lithium hydroxide to give the desired carboxylic acid (10-3). ¹ H-NMR (CDCl ₃ ): δ 8.75 (s, 1H), 7.43 (d, 1H), 7.18 (d, 1H), 4.21 (t, 2H), 2.85 ( t, 3H), 1.4 (s, 9H).

3- [1-cyano-3-oxo-6-2- (piperidin-4-yl) ethyl carbamoyl) -2,3-dihydro-imidazo [1,5-a] pyridin-2-yl!] - propionic acid - a compound of formula (10-4)

To a solution of 10-3 (170 mg, 0.51 mmol) in dichloromethane (10 mL) was added triethylamine (71 mL, 0.51 mmol), HOBT (69.3 mg, 0.51 mmol), 6 mg (0.52 mmol) of EDC and 117.2 mg (0.51 mmol) of compound 2-4. The reaction mixture was stirred under nitrogen for 18 hours, then washed successively with 10% 10% citric acid solution, 10 ml water and 10 ml saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. Chromatography of the residue gave a yellow precipitate (215 mg, 0.43 mmol). This material is then treated with trifluoroacetic acid in dichloromethane to cleave the protecting group to give the trifluoroacetic acid salt of compound (10-4) as a yellow solid, mp 173 ° C.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 1.75 (s, 1H), 7.43 (t, 1H), 7.31 (d, 1H), 7.15 (d, 1H) , 4.23 (t, 2H), 3.41 (t, 2H), 3.23 (d, 2H), 2.83 (m, 2H), 1.85 (d, 2H), 1.53 ( m, 2H), 1.41 (m, 1H), 1.32 (m, 2H).

The compound of formula (11-2) can be prepared as shown in Scheme 27.

-64Methyl [4,5,6,7-tetrahydro-4-oxo-5- [3- (tert-butylpropionyl)] pyrazolo [1,5-a] pyrazin-2-yl] carboxylate (11 Compound of formula (-1)

1.4 g (4.8 mmol) of compound 3-2, 0.90 g (5 mmol) of β-alanine tert-butyl ester hydrochloride and 0.78 g (5.28 mmol) of potassium A mixture of carbonate in acetonitrile (150 mL) was heated at reflux under nitrogen for 4.5 hours, then cooled, filtered and concentrated under reduced pressure. The resulting yellow residue was purified by chromatography on silica gel using dichloromethane (2% methanol) as eluent. This affords the diester (11-1) as a colorless glass.

¹ H-NMR (CDCl 3): δ 7.31 (s, 1H), 4.48 (t, 2H), 3.93 (s, 3H), 3.61 (t, 2H), 2.71 (t .

2H), 2.35 (t, 2H), 1.23 (s, 9H).

3- [4,5,6,7-tetrahydro-4-oxo-2- / 2- (piperidin-4-yl) ethylcarbamoyl / -pyrazolo [1,5-a] pyrazin-5-yl] - Propionic acid - Compound 11-2

To a solution of compound (11-1) (1.0 g, 3.1 mmol) in methanol (10 mL) was added a solution of lithium hydroxide (145 mg, 3.41 mmol) in water (10 mL) and the resulting mixture was heated to 60 ° C. Heat for 2.5 hours. The mixture was cooled and the solvent removed under reduced pressure. The residue was acidified with 10% citric acid and extracted twice with 100 ml of dichloromethane each time. The combined organic extracts were washed with water, dried and evaporated to give the desired acid as a colorless glass.

¹ H-NMR (CDCl 3): δ 7.21 (s, 1H), 4.48 (t, 2H), 3.63 (t, 2H), 2.71 (t, 2H), 2.32 (t .

2H), 1.23 (s, 9H).

-65Of this acid, 500 mg (1.62 mmol) was dissolved in 10 mL of dichloromethane, followed by 220 mg (1.62 mmol) of HOBt, 309 mg (1.62 mmol) of EDC and 356 2-4 mg (1.63 mmol) of compound was added. The reaction mixture was stirred under nitrogen for 16 hours and then washed successively with 10 ml of 10% citric acid solution, 10 ml of water and 10 ml of saturated sodium chloride solution. After drying over anhydrous sodium sulfate and evaporation, the residue obtained is chromatographed to give a colorless foam. This is then deprotected in ethyl acetate with hydrogen chloride to give the hydrochloride salt of compound (11-2) as a white solid, m.p. 192-194 ° C.

¹ H-NMR (300 MHz, DMSO-d 6): δ 9.0 (broad, s, 1H), 8.35 (m, 1H), 6.99 (s, 1H),

4.38 (t, J = 6.0 Hz, 2H), 3.83 (t, J = 5.5 Hz, 2H), 3.64 (t, J = 7.1 Hz, 2H),

3.30-3.1 (m, 4H), 2.85-2.65 (q, 2H), 2.54 (t, J = 7.1 Hz, 2H), 1.85 -1.75 ( s, broad, 2H), 1.60-1.20 (overlapping m, 5H).

Compound 12-4 can be prepared as shown in Scheme 28.

2- (4-Pyridyl) ethylamine (12-1) was charged with a solution of ammonium chloride in water (200 mL) and 4-vinylpyridine (56.4 mL, 0.52 mol) was added and ml of methanol. The resulting mixture was then heated at 60 ° C for 18 hours and then cooled to 0 ° C in an ice bath and made basic by the addition of 30% sodium hydroxide solution. The basic solution was then extracted with dichloromethane (5 x 100 mL), and the combined extracts were dried and evaporated. Vacuum distillation of the residue gave 12-1 as a colorless liquid.

-66 ^1H-NMR (300 MHz, _CDCl3): δ 8.53 (d, J = 6.1 Hz, 2H), 7.25 (d, J = 6.1 Hz, 2H);

3.02 (t, 2H), 2.77 (t, 2H), 1.4 (broad s, 2H).

Methyl [4,5,6,7-tetrahydro-4-oxo-5- [2- (pyridin-4-yl) ethyl] pyrazolo [1,5-a] pyrazin-2-yl] carboxylate - ( 12-2)

A solution of 1.4 g (4.8 mmol) of 3-2, 0.645 g (5.28 mmol) of 12-1 and 0.78 g (5.28 mmol) of potassium carbonate in 150 mL of acetonitrile was added. After refluxing for 4.5 hours, the mixture was cooled, filtered and concentrated under reduced pressure. The resulting yellow residue was redissolved in 50 ml of dimethylformamide and 200 mg of sodium hydride in 60% by weight mineral oil dispersion was added. The reaction mixture was heated at 90 ° C for 3 hours and then concentrated under reduced pressure. The residue was purified by silica gel chromatography using 20% methanol in dichloromethane as eluent. This gave compound 12-2 as a pale yellow glass (0.91 g, 3.0 mmol, 68%).

¹ H-NMR (CDCl 3): δ 8.32 (d, 2H), 7.52 (d, 2H), 7.34 (s, 1H), 4.48 (t, 2H), 3.91 (s) , 3H), 3.61 (t, 2H), 2.71 (t, 2H), 2.35 (t, 2H).

Tert-Butyl 2 (S) - [(p-toluenesulfonyl) amino] -3- [4,5,6,7-tetrahydro-4-oxo-5- [2- (4-pyridyl) ethyl] -pyrazolo [1,5-a] pyrazin-2-yl] carboxylate - Compound 12-3

To a solution of 12-2 (910 mg, 3.0 mmol) in methanol (10 mL) was added a solution of lithium hydroxide (130 mg, 3.05 mmol) in water (10 mL) and the resulting mixture was heated at 60 ° C for 2 hours. Heat for 5 hours. After cooling, the solvent was removed under reduced pressure. The resulting residue was subjected to ion exchange chromatography on Dowex-50W.

-67 further, giving the desired acid as an off-white precipitate, m.p. 187 ° C.

300 mg (0.78 mmol) of this acid are suspended in 50 ml of anhydrous dimethylformamide, and 293 mg (81 mmol) of A-8, 150 mg (0.78 mmol) of EDC are added. HOBt (105 mg, 0.78 mmol) and N-methylmorpholine (87 mL, 0.78 mmol) were added. The resulting clear solution was stirred at 25 ° C for 19 hours and then diluted with 100 mL of ethyl acetate followed by 25 mL of saturated sodium bicarbonate, 25 mL of water, and 25 mL of saturated sodium chloride. washed, dried over anhydrous sodium sulfate and evaporated to give 12-3.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.63 (d, 2H), 7.80 (d, 2H), 7.58 (d, 2H), 7.29 (t, 1H) , 6.93 (s, 1H), 5.95 (d, 2H), 4.40 (t, 2H), 4.08 (m, 1H), 3.86 - 3.74 (m, 4H), 3.35-3.20 (m, 2H), 3.10 (t, 2H), 1.30 (s, 9H).

2 (S) - [(p-toluene-sulfonyl) amino] -3- [4,5,6,7-tetrahydro-4-oxo-5- [2- (4-pyridyl) ethyl] pyrazolo [ 1,5-a] pyrazin-2-yl] carboxylic acid · TFA salt (12-4)

The compound of formula (12-3) is deprotected with trifluoroacetic acid in dichloromethane and purified by reverse phase chromatography. The compound 12-4 is thus obtained in the form of the trifluoroacetate salt, m.p. 182-185 ° C.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.72 (br d, 2H), 8.19 (t, 1H), 8.00 (d, 1H),

7.81 (d, 2H), 7.58 (d, 2H), 7.19 (d, 2H), 6.85 (s, 1H), 4.40 (t, 2H), 4.01 (m , 1H), 3.83-3.76 (m, 4H), 3.48 (m, 1H), 3.26 (m, 1H), 3.10 (t, 2H).

68Α (13-4) can be prepared as shown in Scheme 29.

Methyl 5,6,7,8-tetrahydro-4-oxo-5- [3- (cyanophenyl) methyl-4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl] carboxylate - a compound of Formula 13-1

A solution of 5- (3) (3.02 g, 14.5 mmol) in anhydrous dimethylformamide (60 mL) was cooled to 0 ° C and sodium hydride (636 mg, 15.98 mmol) was added at 60% w / w. in the form of a mineral oil dispersion. The resulting mixture was stirred at 0 ° C for 1.5 hours and a solution of 3.11 g (15.89 mmol) of 3-cyanobenzyl bromide in 50 ml of dimethylformamide was added dropwise. The reaction mixture was stirred at 25 ° C for 18 hours and then diluted with ethyl acetate (200 mL). The reaction mixture was washed with water (3 x 100 mL), saturated brine (100 mL), and the organic layer was dried over anhydrous sodium sulfate, filtered and evaporated. The resulting precipitate was recrystallized from dichloromethane / methanol to give compound 13-1 as a white precipitate. ¹ H-NMR (CDCl ₃ ): δ 7.85 (s, 1H), 7.78 (d, J = 8 Hz, 1H), 7.59 (d, J = 8 Hz, 1H),

7.46 (m, 1H), 7.30 (s, 1H), 4.80 (s, 1H), 4.43 (t, J = 8Hz, 2H), 3.89 (s, 3H), 3.38 (t, J = 8Hz, 2H), 2.09 (m, 2H).

Tert-Butyl 2 (S) - [(p-toluenesulfonyl) amino] -3 - [[5,6,7,8-tetrahydro-4-oxo-5 - [(3-cyanophenyl) - methyl] -4H-pyrazolo [1,5-a] [1,4] diazapin-2-yl) carbonyl] aminoZ-propanoate - Compound 13-2

To a solution of 13-1 (1.5 g, 4.04 mmol) in THF (100 mL) was added 1N lithium hydroxide (5.1 mL, 5.1 mmol) and water (100 mL), followed by a mixture of Stir at 25 ° C for 1.5 hours. The tetrahydrofuran is then removed under reduced pressure and the aqueous residue is acidified with 1 N hydrochloric acid. The precipitate formed is filtered off and dried in vacuo. This gave the desired product as a white precipitate. ¹ H-NMR (CDCl 3): δ 7.95 (s, 1H), 7.73 (d, 1H), 7.53 (d, 1H), 7.43 (m, 1H), 7.30 (s) , 1H), 4.85 (s, 2H), 4.43 (t, 2H), 3.31 (t, 2H), 2.08 (m, 2H).

1.0 g (3.23 mmol) of the acid thus obtained is mixed with 1.24 g (3.54 mmol) of A-9 in 100 ml of dichloromethane, 480 mg (3.54 mmol) of HOBt. and 641 mg (3.54 mmol) of EDC. A further 403 µl (3.83 mmol) of N-methylmorpholine were added and the resulting solution was stirred at room temperature for 16 hours. The mixture was washed successively with 100 ml of saturated sodium bicarbonate solution, 100 ml of 10% potassium bisulphite solution and 100 ml of saturated sodium chloride solution, drying over anhydrous sodium sulphate, filtration and evaporation. . The residue thus obtained was purified by chromatography on silica gel using ethyl acetate as the eluent. This gives compound 13-2 as a white precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.73 (d, J = 6.8 Hz, 2H), 7.68 (s, 1H), 7.65 (d, 1H),

7.51 (m, 1H), 7.37 (d, J = 6.8 Hz, 2H), 7.20 (d, 1H), 7.18 (t, 1H), 5.63 (d, J) = 6.5 Hz, 1H), 4.80 (s, 2H), 4.78 (m, 1H), 4.45 (t, 2H), 3.85 (m, 1H), 3.08 (m , 1H), 3.40 (t, 2H), 2.43 (s, 3H), 2.19 (m, 2H), 1.65 (s, 9H). 2 (S) - [(p-toluenesulfonyl) amino] -3 - / [{5,6,7,8-tetrahydro-4-oxo-5 - [(3-cyanophenyl) methyl] - 4H-Pyrazolo [1,5-a] [1,4] diazapin-2-yl) carbonyl] -amino-propanoate (13-3)

A solution of the compound (13-2) in dichloromethane (15 mL) was treated with trifluoroacetic acid (5 mL) and the resulting solution was stirred at 0 ° C for 2.5 h.

And then evaporated to give 13-3 as a colorless precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.73 (d, J = 6.8 Hz, 12H), 7.65 (s, 1H), 7.65 (d,

1H), 7.50 (m, 1H), 7.31 (d, J = 6.8 Hz, 2H), 7.3 (d, 1H), 7.28 (t, 1H), 6.15 ( d, J = 6.5 Hz, 1H), 4.80 (s, 2H), 4.63 (m, 1H), 4.43 (t, 2H), 3.82 (m, 1H), 3, 65 (m, 1H), 3.48 (m, 2H), 2.43 (s, 3H), 2.19 (m, 2H).

2 (S) - [(p-toluenesulfonyl) amino] -3 - / [{5,6,7,8-tetrahydro-4-oxo-5 - [(3-amidinophenyl) methyl] - 4H-Pyrazolo [1,5-a] [1,4] diazapin-2-yl} carbonyl] -amino} -propionic acid - Compound 13-4

Dissolve 400 mg (0.73 mmol) of 13-3 in 10 ml of a 4: 1 mixture of pyridine and triethylamine and saturate with hydrogen sulfide and stir until the nitrile has a high resolution. liquid chromatography (requires 2.5 hours). The excess hydrogen sulfide is then removed from the solution by passing nitrogen gas and the solution is evaporated. The residue was triturated with 1 N hydrochloric acid and filtered to give a yellow precipitate. This was dissolved in acetone (15 mL) and methyl iodide (250 mL) was added. The resulting mixture was then heated at 50 ° C until thioamide was no longer detectable by high performance liquid chromatography (2 hours). The solvent and excess methyl iodide were evaporated and the residue redissolved in methanol containing 144 mg (1.14 mmol) of ammonium carbonate. The resulting solution was then heated at 50 ° C for 12.5 hours and then concentrated. The compound of formula (13-4) is isolated by preparative reverse phase chromatography.

-71 ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 9.38 (s, 2H), 9.17 (s, 2H), 8.19 (t, 1H), 8.16 (d, 2H), 7.78 (s, 1H), 7.75 (m, 2H), 7.63 (m, 1H), 7.60 (d, 2H), 7.21 (d, 2H), 6, 93 (s, 1H), 4.81 (s, 2H), 4.40 (t, 2H), 3.93 (m, 1H), 3.40 (m, 2H), 3.35 (m, 2H) ), 2.23 (s, 3H), 2.13 (m, 2H).

The preparation of compound (14-5) is illustrated in Scheme 30.

Methyl 5,6,7,8-tetrahydro-4-oxo-5- (3-chloropropyl) -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl] carboxylate - ( 14-1

As described for the preparation of 13-1, 2.0 g (9.5 mmol) of 5-3 is alkylated with 1.5 ml (10.5 mmol) of 1-chloro-3-bromopropane to give Yielding compound (14-1) as a white precipitate.

¹ H-NMR (300 MHz, CDCl ₃ ): δ 7.28 (s, 1H), 4.58 (t, 2H), 3.93 (s, 3H), 2.78 (t, 2H), 2 , 68 (t, 2H), 2.46 (t, 2H), 3.27 (m, 2H), 2.18 (m, 2H).

5,6,7,8-Tetrahydro-4-oxo-5- (3-azidopropyl) -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl] carboxylic acid - (14 -2

A solution of 14-1 (909 mg, 3.2 mmol) and sodium azide (620 mg, 9.5 mmol) in dimethylformamide (15 mL) was stirred at room temperature for 36 hours and then diluted with ethyl acetate (50 mL). It was washed three times with 50 ml of water, dried over anhydrous sodium sulfate, filtered and evaporated to give the azide as a white precipitate. This is then hydrolyzed in the usual manner to give 14-2 as a white precipitate.

¹ H-NMR (300 MHz, DMSO-d 6): δ 7.00 (s, 1H), 4.41 (t, 2H), 3.52 (t, 2H), 3.41 (t, 2H), 3.26 (t, 2H), 2.20 (m, 2H), 1.80 (m, 2H).

-72-tert-Butyl 2 (S) - (p-toluenesulfonylamino) -3- [5,6,7,8-tetrahydro-4-oxo-5- (3-aminopropyl) -4H-pyrazole [1,5-a] [1,4] Diazepin-2-yl] carboxyl] amino] -propionate (14-3)

As described for the preparation of 13-2, the acid 14-2 is coupled with A-9 to give the desired product as a white precipitate. This was then dissolved in ethanol and hydrogenated under an atmosphere of hydrogen in the presence of 10% palladium on carbon to give 14-3 as a white precipitate.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.18 (t, 1H), 7.68 (d, 2H), 7.23 (d, 2H), 6.98 (s, 1H) , 4.4 (m, 3H), 3.93 (t, 2H), 3.48-3.2 (m, 6H), 2.78 (t, 2H), 2.43 (s, 3H), 2.69 (m, 2H), 1.86 (m, 2H), 1.08 (s, 9H).

2 (S) - (p-Toluenesulfonylamino) -3- [5,6,7,8-tetrahydro-4-oxo-5- (3-guanidino-propyl) -4 H -pyrazolo [1,5- a] [1,4] Diazepin-2-yl] carbonyl) amino) -propionic acid (14-4) in a solution in mg (0.1 mmol) of (14-3) in 5 mL of dimethylformamide 90 µl (0.5 mmol) DIPEA and 30 mg (0.5 mmol) are added

3,5-dimethylpyrazole-1-carboxamidine and the resulting mixture was stirred at 80 ° C for 12 hours. The reaction mixture was evaporated and the residue was purified by chromatography on neutral alumina using 80: 20: 1 by volume dichloromethane: methanol: ammonium hydroxide as eluent. This gave the desired product as a white precipitate. This is then treated with trifluoroacetic acid for the deprotection in the usual manner and then subjected to preparative reverse-phase chromatography. This gives compound 14-4 as a white precipitate.

• ♦ · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

¹ H-NMR (300 MHz, D ₂ O): δ 8.2 (t, 1H), 7.58 (s, 2H), 7.18 (d, 2H), 4.38 (t, 2H) )

3.51 (m, 5H), 3.45 (t, 2H), 3.2 (m, 1H), 3.18 (m, 2H), 2.10 (s, 3H), 2.08 (m , 2H), 1.8 (m, 2H).

2 (S) - (p-toluenesulfonyl) -3- [5,6,7,8-tetrahydro-4-oxo-5- [3- [N- (imidazolin-2-yl) amino] -propyl] -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl] carboxylaminopropionic acid - Compound 14-5

As described for the preparation of compound (14-4), the solution of compound (14-3) was reacted with 2- (methylthio) -2-imidazoline hydroiodide and the resulting crude product was treated with trifluoroacetic acid for deprotection. . Thus, compound 14-5 can be isolated by preparative reverse phase chromatography.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ 8.20 (t, 1H), 8.10 (d, 2H), 7.60 (d, 2H), 7.21 (d, 3H) , 6.86 (s, 1H), 4.46 (t, 2H), 4.01 (m, 2H), 3.8 - 3.5 (overlapping m,

8H), 2.23 (s, 3H), 2.10 (t, 2H), 1.80 (t, 2H).

The compound of formula (15-2) can be prepared as shown in Scheme 31.

2 (S) - [(p-toluenesulfonyl) amino] -3 - / [{5,6,7,8-tetrahydro-4-oxo-5- [2- (N-BOC-piperidin-4- yl) ethyl] -4H-pyrazolo [1,5-a] [1,4] diazepin-2-yl) carbonyl] aminoZ-propionic acid - Compound 15-1

A solution of 5.0 g (12.3 mmol) of 5-6 in 150 mL of tetrahydrofuran was cooled to 0-10 ° C and added via syringe

N-methylmorpholine (2.11 mL, 19.2 mmol). After stirring for 20 minutes, 2.38 ml (18.2 mmol) of isobutyl chloroformate was added dropwise via syringe and the resulting solution was stirred for 0.5 hour to give the desired mixed anhydride.

• · ·

In a 500 mL round bottom flask equipped with a magnetic stir bar, 7.00 g (27.1 mmol) of A-7, 125 mL of tetrahydrofuran and 4.71 mL (27.1 mmol) of diisopropylethylamine are mixed and water is added in portions until a clear solution is obtained. This solution was then cooled in an ice bath, and the mixed anhydride suspension prepared as described in the previous paragraph was added in one portion with vigorous stirring. After stirring for 20 minutes, the reaction mixture was concentrated to remove tetrahydrofuran. The residual aqueous material was acidified with 10% potassium bisulfate solution and the resulting precipitate was filtered to give a white precipitate. This was then subjected to flash chromatography on a silica gel column (14.6-25.4 mm) supplied by EM Science with dichloromethane, methanol and ammonium hydroxide 98: 2: 0.2: 95: 5: 0.5, 90: 10: 1 and finally 85: 15:: 1.5 (v / v). This gives pure 15-1 as a white precipitate.

¹ H-NMR (DMSO-d ₆ ): δ 8.23 (q, J = 3.40 Hz, 1H), 7.64 (d, J = 8.20 Hz, 2H), 7.32 (d, J = 8.20 Hz), 7.2 - 7.0 (broad, 1H), 6.86 (s, 1H), 4.36 (t, J = 6.70 Hz, 2H), 3.89 ( d, broad, J = 12.21 Hz, 2H), 3.59 (m, 1H), 3.47 (t, J = 7.08 Hz, 2H), 3.5 - 3.1 (m, broad , 5H, H ₂ O), 2.8-2.6 (broad, 2H), 2.33 (s, 3H),

2.17 (t, J = 6.47 Hz, 2H), 1.66 (d, broad, J = 11.97 Hz, 2H), 1.55 - 1.45 (m, broad, 3H), 1 , 37 (s, 9H), 1.1-0.9 (m, broad, 2H).

-752 (S) - [(p-toluenesulfonyl) amino] -3 - / [{5,6,7,8-tetrahydro-4-oxo-5- [2- (piperidin-4-yl) - ethyl] -4H-pyrazolo [1,5-a] _[1L 4] diazepin-2-yl} carbonyl] amino / propionic acid - (15-2) the compound formula

Weigh into a 1 liter round bottom flask equipped with a magnetic stirrer

7.42 g (11.48 mmol) of compound 15-1 are added followed by methylene chloride and the mixture is cooled to 0-5 ° C. The resulting suspension is then bubbled through with gaseous hydrogen chloride while stirring. After about 2 minutes the precipitate goes into solution and a second precipitate soon forms. After gas was bubbled through the suspension for an additional 5 minutes, the reaction vessel was warmed to room temperature. After 30 minutes, the contents of the reaction vessel were evaporated. The resulting white precipitate is the hydrochloride salt of compound (15-2), which has a purity of greater than 99% as determined by HPLC.

The hydrochloride salt of compound (15-2) is subjected to ion exchange chromatography at 110 g (4.11 molar equivalent / g) from Dowex 50X8-200 ion exchange resin. The resin was prepared by washing with 500 ml water, 500 ml methanol, 500 ml water, 500 ml 6N hydrochloric acid and 500 ml water. At this time, the eluent had a pH of 7. The hydrochloride salt was dissolved in 30 ml of water and applied to the top of the column. The column was eluted with water. The pH of the eluent becomes strongly acidic. When the pH of the eluent returns to 7, the column was eluted with a 50:25:25 mixture of ammonium hydroxide, acetonitrile and water.

Elute with 1.5 liters. The ultraviolet light-containing fractions were pooled and concentrated under high vacuum. The resulting white foam was dried under high vacuum for 8 hours to give 15-2.

-76 ^1H-NMR _(DMSO-d6): δ 9.0 to 8.5 (br, 1H), 8.19 to 8.16 (m, 1H), 7.67 (d,

J = 8.18 Hz, 2H), 7.32 (d, J = 8.18 Hz, 2H), 6.89 (s, 1H), 4.38 (t,

J = 6.84 Hz, 2H), 3.75 - 3.65 (m, broad, 1H), 3.46 (t, broad, 2H), 3.5 - 3.1 (m, broad, 8H) H ₂ O), 2.77 (t, broad, J = 11.36 Hz, 2H), 2.35 (s, 3H), 2.17 (t, J = 6.47 Hz, 2H), 1, 80 (d, broad, J = 12.7 Hz, 2H), 1.53 -

1.42 (m, broad, 3H), 1.33-1.24 (m, broad, 2H).

Using the methods described above, in particular those depicted in Schemes 12 and 15, the compounds listed in Table 1 can be prepared.

TABLE 1 (LXXXVII)

R mp (° C) Salt form (LXXXIX) 115-120 TFA (XC) 110-120 TFA (XCI) 160-165 double ion (XCII) 195-198 double ion 180-188 HCI (XCIII) 150-156 HCI (XCIV) 195-200 double ion (XCV) 121-124 TFA (XCVI) 210-212 TFA 85-95 TFA (XCVI) 175180 TFA (XCVII) 115-120 HCI (XCVIII) 78-80 TFA (XCIX) HCI (C) 200 (decomposes) HCI so _2- nh ₂ 210 (decomposes) HCI

Further compounds may be prepared in an analogous manner to the compounds described in the Examples above. These additional compounds are listed in Tables 2-4.

TABLE 2 (Cl)

R THE B o p. (° C) (CII) H H 145-152 (CIH) H H 168-170 (CIII) ch ₃ H 164-167 (CIV) H H 110-135 CN H H 180-188 (CV) H NHSO2C4H9 156-160 (CIII) H NHSO2C4H9 134-140 H NHSO2C4H9 110-125 (CVI) H NHSO2C4H9 130-135 (CV) H (CVIII) 185-190

TABLE 3 (CIX)

n m mp (° C) 2 0 110-115 2 1 115-120 1 1 121-123 0 2 135-141 0 1 140-145

TABLE 4 (CX)

R Amide's stereochemistry of H _e space- NH * chemical shift section (PP-m). H cis 6.35 H trans 7.38 NHSO ₂ C ₄ H ₉ cis 6.31 nhso ₂ c ₄ h ₉ trans 7.29

He refers to the 6-position hydrogen of the bicyclic structure.

Table 5 (CXII)

Ri r ₂ mp (° C) H SO ₂ C ₄ H ₉ 110-116 H (CVIII) 186 ch ₃ (CVIII) 164-171 H (XCVI) 164-170 H (CXIII) 135-140 H (CXIII) not defined H (CXV) 173-174 H (XCVI II) 185-189 H H 178-179

TABLE 6 (CXVI)

n Ri r ₂ mp (° C) 1 H H 248-249 1 H (CXVII) 175-180 1 ch ₃ H 118-122 1 ch ₃ (CXVII) 155-160 2 Ph H 137-139 2 Ph (CXVII) 185 - 188 2 H (CXVII) 192-194 2 H NHCO ₂ CH ₂ Ph 176-178 2 H NHSO ₂ C ₃ H ₇ 168-170 2 H nhso ₂ c ₂ h ₅ 173-174

Claims (11)

PATENT CLAIMS CLAIMS 1. Compounds of formula (I) or (II) and pharmaceutically acceptable salts thereof Q is a group of formula (IV) or (V) or a group of formula (VI) or Q is a 4- to 9-membered mono- or bicyclic ring group selected from nitrogen, oxygen and sulfur atoms 1, 2 or 3 containing a heteroatom and optionally substituted with R ⁸ as defined below; AB is a condensed ring system containing adjacent carbon and nitrogen atoms in which A is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; and B is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; R ¹ is hydrogen, 1-4 alkyl, or -N ^(R8) 2, -N ^(R8) SO2R ^7, -NR ⁸ CO ₂ R ^7, -NR ⁸ C (O) R ^7, -NR ⁸ C (O) N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁸ ) C (O) OR ⁷ , -C (O) N (R ⁷ ) ₂ or R ⁶ as defined below a cyclic ring; R ² is hydrogen or C 1-4 straight or branched alkyl, arylC 1-4 alkyl or aryl; R ⁴ is hydrogen, C 1 -C 4 straight or branched chain or cyclic alkyl, or C 1 -C 4 alkenyl; ········································································································································································································································ Management -82R ⁵ is a methyl group, -CH (CH ₂ ) n -, a bond or, when R ^{5 is} adjacent to N (R ⁴ ) - a group of formula VII; R ^e is carboxyl, hydroxymethyl, tetrazolyl, acylsulfonamido, -C (O) N (R ⁷ ) ₂ , -CO ₂ R ⁹ , or (VIII) or R ⁶ R ¹ with a substituent forms a group of formula (IX) or (X); y is oxygen or sulfur; R ⁷ is hydrogen, or C 1-4 straight or branched, optionally substituted alkyl, straight or branched lower alkenyl, aryl (C 1-4) alkyl, substituted aryl, or 5- or 6-membered, Heteroaryl radicals containing 1, 2 or 3 nitrogen sulfur or oxygen atoms as heteroatoms; wherein the substituted alkyl group is substituted with hydroxy or C 1-4 alkoxy and the substituted aryl group has 1, 2 or 3 substituents selected from halogen, C 1-4 alkoxy, hydroxy or C 1-4 alkyl; R ⁸ is hydrogen or C 1-4 straight or branched chain alkyl; R ⁹ is hydrogen or C 1-4 alkyl or aryl; n is 0, 1, 2, 3, 4, 5, 6 or 7; ri is 0, 1, 2 or 3; and a is a group of formula XI or a chemical bond. Compounds of formula (I) or (II) according to claim 1, and also their racemates, their racemic mixtures, their enantiomers and their diastereomers, characterized in that: -83Q represents (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI), (XXII), (XXIII) , (XXIV), (XXV), (XXVI), (XXVII), (XXVIII), (XXIX) or (XXX); n is 0, 1, 2, 3, 4, 5, 6 or 7; ri is 0, 1, 2 or 3; R ⁴ is hydrogen, straight or branched or cyclic alkyl of 1 to 4 carbon atoms or alkenyl of 1 to 4 carbon atoms; ^R5 is CH, -CH _{(CH2) n-} or a bond; R ² is hydrogen, straight or branched (C 1 -C 4) alkyl, aryl (C 1 -C 4) alkyl, or aryl; ^R1 is hydrogen, C1 -C4 alkyl or -N ^(R8) 2, -N ^(R8) SO2R ^7, -NR ⁸ CO ₂ R ^7, -NR ⁸ C (O) R ^7, -NR ⁸ C (O) N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁷ ) R ⁸ , -N (R ⁸ ) SO 2 N (R ⁸ ) C (O) OR ⁷ , -C (O) N (R ⁷ ) ₂ or R ⁶ ; R ⁶ is carboxyl, hydroxymethyl, tetrazolyl, acylsulfonamido, -C (O) N (R ⁷ ) ₂ , -CO ₂ R ⁹ or (VIII) or together with R ^{1 is} cyclic; R ¹ and R ⁶ together form a cyclic group of formula (IX) or (X); y is oxygen or sulfur; R ⁷ is hydrogen, or C 1 -C 4 straight or branched, optionally substituted alkyl, straight or branched lower alkenyl, aryl (C 1 -C 4) alkyl, substituted aryl, or 5- or 6-membered, 1, 2 or 3 nitrogen, sulfur or oxygen atom as a 84-week • a heteroatom · · · ·· ♦ ♦ • ♦ * · _Λ · w ·· ·· ·· ··· ·· ·· ·· · ♦ · "-Aryl"; wherein the substituted alkyl group is substituted by hydroxy or C 1-4 alkoxy and the substituted aryl group has 1, 2 or 3 substituents selected from halogen, C 1-4 alkoxy, hydroxy-alkyl, and C 1-4; R ⁸ is hydrogen or C 1-4 straight or branched chain alkyl; R ⁹ is hydrogen or C 1-4 alkyl or aryl; a is a group of formula XI or a bond; A is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; and B is a 5-, 6- or 7-membered saturated or unsaturated ring containing 1, 2 or 3 heteroatoms selected from oxygen, sulfur and nitrogen; whereas A and B together form a fused ring system in which adjacent carbon and nitrogen atoms are common. Compounds of formula (I) according to claim 2, characterized in that AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH ₂ - or (XXXII) - or (XXXIII) ) - in which X represents a nitrogen atom or a group -CR ³ , in which R ³ ···· 4 «» • · · * <» Cyano -85lentése -C (O) N ^{(R7) R8,} (XXXIV), (XXXV) or (XXXVI), a group of formula - or (XXXIX), (XL) or (XLI) R. Compounds of formula (I) according to claim 3, characterized in that AB is a group of formula (XLII), in which V is a nitrogen or methine group and D is an ethylene group or a -CH ₂ C (R ⁴ ) ₂ CH ₂ - group. Compounds of formula (II) according to claim 2, characterized in that AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH ₂ - or (XXXII) - or (XXXIII) ) - in which X is nitrogen or -CR ³ and in which R ³ is cyano, -C (O) N (R ⁷ ) R ⁸ , (XXXIV) or (XXXV) - or a group of formula (XL), (XLI) or (XLV), wherein in the latter, y ³ is oxygen or two hydrogen. Compounds of formula (II) according to claim 5, characterized in that a is a compound of formula (XI); and AB is a group of formula (XXXI) wherein V is nitrogen or -CR ⁷ and D is methylene, ethylene, -CH 2 C (R ⁷ ) 2 CH ₂ - or (XXXII) or (XXXIII) - where X is nitrogen -86 atoms or -CR ³ , wherein R ³ is cyano, -C (O) N (R ⁷ ) R ⁸ , (XXXIV) or (XXXV). Compounds of formula (II) according to claim 5, characterized in that a is a chemical bond; and AB is a group of formula (XL), (XLI) or (XLV), and in the latter, y ³ is oxygen or two hydrogen. The compound of formula (I) or (II) according to claim 2, which is (XLVI), (XLVII), (XLVIII), (XLIX), (L), (Ll), (III), (Lil). , (Lili), (LIV), (LV), (LVI), (LVII), (LVIII), (LIX), (LX), (LXI), (LXII), (LXIII), (LXIV), ( LXV), (LXVI), (LXVII), (LXVIII), (LXIX), (LXX), (LXXI), (LXXII), (LXXIII), (LXXIV), (LXXV), (LXXVI), (LXXVII) , LXXVI II, LXXIX and LXXX. A compound of formula (I) or (II) according to claim 2, which is a compound of formula (LXXXI), (LXXXII), (LXXXIII), (LXXXIV) and (LXXXV). Compounds of formula (I) or (II) according to claim 2 for inhibiting fibrinogen binding to platelets, inhibiting platelet aggregation, treating thrombus formation or embolism, or preventing thrombus formation or embolism. 11. A pharmaceutical composition for inhibiting fibrinogen binding to platelets in mammals, inhibiting platelet aggregation, treating thrombus formation or embolism, or preventing thrombus formation or embolism, comprising a compound according to claim 2 and a pharmaceutically acceptable carrier.