SI21271A - Fibrinogen receptor antagonists - Google Patents

Abstract

The invention refers to fibrinogen receptor antagonists having the general formula (I) in which substituents have meanings given in the description.

Description

The invention relates to the pharmaceutical industry and relates to novel heterocyclic analogues of the Arg-Gly-Asp tripeptide that modulate cell adhesion and prevent the binding of fibrinogen and other proteins to platelets via the integrin GPIIb / lll receptor. They prevent platelet aggregation and thus blood clot formation. The invention also relates to processes for the preparation of new compounds, to pharmaceutical products containing them, and to use in therapy.

A technical problem

Pathological changes in blood clotting are the cause of many cardiovascular diseases, which are among the most common diseases of today in the developed world. A common problem affecting patients with cardiovascular disease is increased platelet aggregation and subsequent clot formation. This is due to the complexity of the physiological mechanisms that control the flow of blood through the cardiovascular system and allow it to clot upon contact with nonendothelial surfaces at the sites of vascular injury. In the normal state, there is a homeostatic balance between thrombosis and hemorrhage, which permits physiological fibrinolysis of intravascular plugs while preventing excessive pathological fibrinogenolysis. The agents that influence this complex system of regulation of blood flow vary greatly in mechanism of action, all of which affect the balance between procoagulant and anticoagulant reactions. The effectiveness of these compounds is closely related to their toxicity; the desired therapeutic effect of a compound with anticoagulant action may be rapidly accompanied by bleeding due to overdose of the active substance. Many cardiovascular complications, such as arterial and venous thrombosis, stroke, heart attack, and pulmonary embolism, arise as a direct result of pathological changes in the structure of the vascular wall that trigger the mechanisms of blood clotting. Therefore, the compounds that influence these mechanisms play the most important role in therapy, and especially in the prevention of cardiovascular disease.

The state of the art

Anticoagulants, which are currently used in therapy, have many side effects (JG Hardman, Gilman AG, Limbird LE: Goodman &Gilman's The Pharmacoiogical Basis of Therapeutics, 9 ^th ed., McGraw-Hill, New York, 1996: 1341-59) . Also, the vast majority of active substances need to be administered intravenously. Therefore, in the field of anticoagulants, especially in recent decades, attempts have been made to synthesize new, more specific agents that would be acceptable for oral therapy, which would reversibly and specifically inhibit platelet aggregation, regardless of the type of agonist, but would not significantly affect other normal physiological processes. in the body.

The fibrinogen receptor GPIIb / llla belongs to the beta 3 integrin receptor family and is the best known and studied integrin receptor (Cox D, Aoki T, Seki J, et al .: The Pharmacology of the Integrins, Med. Res. Rev., 1994, 14 : 195-228; Eldred CD, Judkins BD: Fibrinogen Receptor Antagonists: Design and Ciinical Applications, Progress in Medical Chemistry, Elsevier, eds. King FD and Oxford AW, Amsterdam, 1999, 36: 29-90). This receptor subfamily is characterized by a tripeptide recognition sequence of ligands composed of arginine, glycine, and aspartic acid (RGD). The fibrinogen receptor is located on the membrane of platelets that it is characterized by. Like other integrins, it consists of a complex of two trans me m defers of gl glio co proteins (alpha- and beta-subunits) with long extracellular globular domains and a short cytoplasmic moiety. Extracellular domains contain binding sites for ligands and magnesium and calcium ions, which are necessary for maintaining the integrity of the molecule, but are also essential for ligand binding.

Under normal conditions, the GPIIb / llla receptor is dormant, fibrinogen binding sites are not exposed, and plasma fibrinogen is not bound to it. Fibrinogen binding sites at the GPIIb / llla receptor are exposed after complex intracellular signaling following platelet activation. Despite the presence of other ligands, the fibrinogen is mainly bound to the GPIIb / llla receptor. The identification of the RGD sequence as the smallest fraction of fibrinogen that allows binding to the fibrinogen receptor provided a starting point for fibrinogen receptor antagonist design. From the structure of RGD and some analogues, it was found that the guanidine group Arg and the beta-carboxyl group Asp are essential for antagonistic action and that the spatial distance between these groups must range between 1.2-1.8 nm.

Because it is a peptide structure, we encounter well-known limitations of its therapeutic utility, such as low bioavailability, poor metabolic stability and indiscriminacy, which cause undesirable side effects. Therefore, the modification and replacement of individual parts sought to improve the properties of the well compound. The development of the firinogen receptor antagonists has gone in the search for metabolically stable and orally effective and selective compounds, from simple linear peptides to non-peptide compounds - peptidomimetics (Eldred CD, Judkins BD: Fibrinogen Receptor Antagonists: Design and Clinical Applications, Progress in Medical Chemistry, Elsevier, eds King FD and Oxford AW, Amsterdam, 1999, 36: 29-90; Ojima I, Chakravarty S, Dong Q: Antithrombotic Agents: From RGD to Peptide Mimetics, Bioorganic & Medical Chemistry, 1995, 3, 4: 337-60 Duggan ME, Duong LT, Fisher JE, et al: Nonpeptide α _ν β3 Antagonists 1. Transformation of a Potent, Integrin-Selective GPIIb / llla Antagonist into a Potent α _ν β3 Antagonist, J. Med. Chem., 2000 , 43: 3736-45).

Initially, the researchers synthesized simple acyclic peptides containing the RGD sequence. The synthesized compounds had a low potency and a very short duration of action, and were also indiscriminate to other integrin receptors.

This led to the synthesis of cyclic peptides. The cyclic structure is more rigid and contributes to the greater selectivity of the binding. The approaches for synthesis of cyclic peptides containing the RGD sequence in their structure were different: binding of the peptide chain to the cyclic form via the amino acid cysteine with a disulfide bridge, binding of the peptide chain to the cyclic form via a non-peptide linkage, incorporation of the RGD sequence into the non-peptide linkage. The additional binding of lipophilic amino acids to the RGD sequence was found to increase the potency. Despite the improved selectivity, the metabolic stability of cyclic peptides is not satisfactory.

In order to improve the poor bioavailability of peptide analogues of RGD, nonpeptide analogues were synthesized on the basis of the peptide compound of water. Benzamidine or various cyclic amines have often been used as mimetics of the arginine portion of the RGD sequence. Semi-peptide structures with a benzamidine group had a favorable potency and were selective for the GPIIb / lll receptor. Many non-peptide analogues have been developed, such as e.g. lamifiban, sibrafiban, eptifibatide, torofibanic chloride, abciximab, lefradafiban.

The optimal spatial structure of the GPlIb / llla receptor binding molecules has been attempted by introducing different non-peptide spacers. Many compounds were shown to be effective, which included the benzofuran, thiazole, isoxazoline, indole, benzodiazepine, benzimidazole, benzopyran ring and many others as a spacer.

The bioavailability of fibrinogen antagonists after oral administration is very low due to their zwitterionic structure. Their volume of distribution and half-life are also small, which is reflected in the short duration of the pharmacodynamic effect. The half-life of fibrinogen antagonists can be extended by technological modifications (eg intravenous administration of microspheres). Bioavailability can be increased by the preparation of prodrugs. Non-peptide RGD analogs indicate a new direction for the search for fibrinogen antagonists in the form of small and rigid structures with greater bioavailability after oral administration.

The therapeutic use of fibrinogen receptor antagonists due to their mechanism of action always poses a varying degree of risk of bleeding and related problems, which is a disadvantage of all anti-aggregating agents. Despite the aforementioned disadvantages of this group of anticoagulants, their advantage is primarily in their extremely favorable site of action in the aggregation process, since they inhibit all pathways that cause platelet plug formation.

Description of the invention with embodiments

The subject of the invention are novel compounds of general formula (I):

where they mean:

A is an O, S, N or methylene group,

B is C or N,

E and D are H or a chain of the following structure:

Xa- (CH ₂ ) _n -Y-, where:

where

R represents H, OH, COORi or COR1,

R4 represents O or S and

R ₅ H is an alkyl branched or unbranched C1-C7 chain, but is a CO or 5-7 membered alicyclic or aromatic ring system which, in addition to C, may contain 1 or 2 heteroatoms (O, N). The nitrogen atom in these structures can be substituted with H or with group X, and the benzene ring system at the p- or m-site is substituted with group X, n is 0-4,

Y is O, -NH-CO-, -CO-NH-, NH-CO-NH-, -NH-, -NH-CH ₂ -, -CH2-NH-, -CH2-NH-CO-, -CO -NH-CH 2 -;

wherein one of E and D is always H and the other is a chain;

R1 is H or an alkyl branched or unbranched C1-C4 chain,

R ₂ is HaliCOORi, -CONHR ₆ , -NHCOR7, -CH ₂ -COORi, -CH ₂ -CONHR ₆ , -O-CH ₂ -COORi, -O-CH ₂ -CO NHRe,

CO., LTD

o.

wherein R 6 is a group of the formula wherein R _{8 is} H, NH-SO 2 R ₁₀ . branched or unbranched alkyl chain of C1-C4 atoms, benzyl group,

Rg is H or a branched or unbranched C1-C4 alkyl chain,

Rio is an aliphatic group of C 1 -C 6 or wherein R 11 is -CH 3, -NH 2 or -NHCOCl · ^

R7 is a benzene ring which is substituted at the p- or m-position with a group X,

R ₃ is H or a branched or unbranched alkyl group of C 1 -C 4 atoms, - (CH 2) _n , -COOR 1 or

x

The compounds described in the invention have one or more stereogenic centers at which the absolute configuration may be R or S and may occur in the form of racemates, racemic mixtures, pure enantiomers, mixtures of diastereoisomers or in the form of pure diastreomers.

The invention also relates to pure diastereomers, mixtures of diastereomers, pure enantiomers and mixtures of enantiomers, and to the pharmaceutically acceptable salts of the compounds of formula I. The subject of the invention is also the use of these compounds in therapy and the pharmaceutical compositions containing them. The invention also relates to processes for the preparation of compounds of general formula I.

The novel compounds are fibrinogen receptor antagonists. They inhibit platelet aggregation induced by the binding of fibrinogen to the fibrinogen receptor on platelets. They can be used to prevent and treat arterial thrombosis, ischemic stroke, peripheral arterial disease, acute coronary syndrome, prevent acute heart complications, treat acute myocardial syndrome, and prevent cardiac ischemic complications in patients with surgery. They can be used orally and / or parenterally.

The pharmaceutical products of the present invention may be in the form of injections or oral preparations. In addition to the active substance, they contain various standard additives, depending on the type of use. Pharmaceutical products are prepared by standard procedures. The dose, frequency and method of administration depend on the individual active substance and its pharmacokinetic properties and the patient's condition.

The starting heterocyclic compounds for the preparation of the compounds of the present invention are prepared according to the methods described in the literature.

Compounds 10, 12 were prepared according to the procedure described in the literature (D. R. Shridhar, S. S. Gandhi, K. Srinivasa Rao: Synthesis; 1982, 986-987).

For the removal of the protecting groups of the carboxyl group, commonly known methods are used in the art, and peptide bonds are also formed following the standard procedures described in Bodanszky M., Bodanszky A. The Peptide Synthesis Practice, Springer, Berlin, 1994.

Pharmaceutically acceptable salts of the compounds of formula I are prepared by treating these compounds with acids or bases in suitable organic solvents, as is commonly known in the art.

Evaluation of the biological activity of fibrinogen receptor antagonists

1. Measurement of platelet aggregation

The measurements were performed with the help of the automatic coagulation apparatus Behring Coagulation Timer (BCT, Behring Diagnostics GmbH), whose operation is based on the principle of turbidimetry. The apparatus measured the optical density of the plasma by platelets, which decreased with platelet aggregation. In the platelet plasma sample (PRP), the optical density value was highest before aggregation. On complete aggregation, however, the optical density of PRP was equal to the optical density of platelet poor (PPP).

Work was performed by first inserting plasma, both agonists (ADP, collagen, adrenaline) into the BCT, and the antagonist or appropriate solvent. The apparatus was then dosed with 135 µL of platelet plasma, 15 µL of antagonist or solvent, and 15 µL of agonist in each cuvette. The plasma sample and the antagonist or solvent were incubated for 300 seconds, after which the aggregation was triggered by the apparatus which triggered aggregation. The duration of the entire measurement was 600 seconds. First, we measured the optical density of PPP and the optical density of PRP with the added solvent (saline) without the antagonist, which was then used as a control.

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A PC connected to the BCT plotted a curve that showed the change in optical density as a function of time. At the same time, it displayed the initial and final values of the optical density (mOG unit) and the maximum rate of change of optical density over time (mOG / min).

Aggregation expressed as a percentage was calculated by the formula:

% aggregation ⁼ (OG _{for even} ~ OGfinal / OG _{for even} ^- OG ppp) x 100

Initial: Optical plasma density immediately after agonist addition

OGfinal: Optical plasma density after 10 minutes

OGrpp: Optical density of poor plasma with platelets

For ADP-induced aggregation, the maximum change in optical density was read manually from the curve because the plasma had already disaggregated after 10 minutes.

2. Calculation of aggregation inhibition

The percentage inhibition of aggregation was calculated from% aggregation upon addition of antagonist and% aggregation of control without antagonist by the equation:

% aggregation inhibition = [1 - (% aggregation with antagonist:% control aggregation)] x 100

The percentage inhibition of aggregation rate was calculated from the aggregation rate with the addition of the antagonist and the aggregation rate of the control without the antagonist by the equation:

% aggregation rate inhibition = [1- (mOG / min with antagonist: mOG / min controls)] x 100

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For known antagonists, IC50 was expressed as the arithmetic mean ± standard error of measurements in 2 to 6 different plasmas and for peptidomimetics as the average of measurements in at least two different plasmas.

3. Calculation of IC ₅ o for certain compounds of the invention

We have shown inhibition of aggregation as the concentration of an antagonist that has 50% inhibited aggregation (IC 50 aggregation) or aggregation rate (IC 50 velocity). IC ₅ o values were read from curves showing the percentage inhibition of aggregation with respect to the rate of aggregation as a function of peptidomimetic concentration. IC ₅ o are for some of the compounds which are the subject of the patent application in embodiment 34.

Statistical methods

Platelet aggregation reference values in apparently healthy volunteers were expressed as arithmetic means ± standard deviations and aggregation variability as a coefficient of variability. For known GPIIb / IIIa antagonists, lC ₅ o was expressed as the arithmetic mean ± standard error of measurements in 2 to 6 different plasmas and for peptidomimetics as the average of measurements (arithmetic mean) in at least two different plasmas.

EXECUTIVE EXAMPLES

The invention is explained, but in no way limited by the following embodiments:

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Reaction scheme for the preparation of compound 5:

A: BrCH ₂ COOEt, KF, dimethylformamide, B: BrCH ₂ CH ₂ CH ₂ CH ₂ COOEt, K ₂ CO ₃ , triethylbenzylammonium chloride, MeCN, C: H ₂ , Pd / C, EtOH, D: 4-cyanobenzoyl chloride, Et ₃ N, CHCl ₃ , E: 1. HCl (g), EtOH; 2. Ammonium acetate

Example 1:

7-NITRO-2 / 7-1,4-BENZOXAZIN-3 (4H) -ONE (1)

Weigh 14.74 g (253.7 mmol) of KF into a 100 ml flask, add 75 m of dimethylformamide and stir the resulting suspension. Then add 15.00 g

-1313 (97.3 mmol) of 2-amino-5-nitrophenol and 10.8 ml, (97.3 mmol) of dibromoacetic acid ethyl ester. The reaction mixture was stirred for 15 minutes at room temperature and then started heating on an oil bath at 50-60 ° C. After 6 hours, stop heating and allow the reaction mixture to stir at room temperature overnight. Pour the reaction mixture into 441 ml of a mixture of water and ice. A brown precipitate disappears, which is filtered off by suction and air-dried. Crystallization from ethanol (570 ml) followed. Yield: 10.747 g (56.9%), melting point: 235 239 ° C

IR (KBr, cm-1): 3567,3191, 1697, 1598, 1508, 1391, 1341, 1222, 1043,862, 742, 525

1 H-NMR (300 MHz, DMSO-d 6): δ (ppm) = 4.72 (s, 2H, -OCH ₂ -); 7.06 (d, 1H, J = 8.7 Hz, BO-H); 7.75 (d, 1H, J = 2.3 Hz, BO-H); 7.89 (dd, 1H, J1 = 8.7 Hz, J2 = 2.3 Hz, BO-H); 11.30 (s, 1H, -NH-)

Example 2;

Ethyl 5- (7-nitro-3-oxo-2,3-dihydro-4tf-1,4-benzoxazin-4-yl) pentanoate (2)

To a suspension of 2.50 g (12.9 mmol) of compound 1, 4.46 g (32.2 mmol) of K ₂ CO3, 0.59 g (2.58 mmol) of benzyltriethylammonium chloride and 90 ml of acetonitrile were added 2 while stirring. 55 ml (15.45 mmol) of 5-bromopentanoic acid ethyl ester. The reaction mixture was heated in an oil bath at 60 ° C for three days. After 3 days, 0.61 ml (3.89 mmol) of 5-bromovalerioic acid ethyl ester was added and heated to the next day. Again, a little of 5-5bromopentanoic acid ethyl ester (0.37 ml; 2.33 mmol) was added and heated until the next day. Increase the heating temperature to 75 ° C, add 0.25 ml (1.55 m mol) of 5-bromopentanoic acid ethyl ester and heat to the next

-1414 days. The precipitate is filtered off. The solvent was evaporated under reduced pressure and the residue was dissolved in 250 ml of ethyl acetate. The organic phase was washed successively with 0.1 M HCl (2 χ 50 mL), saturated aqueous NaHCO ₃ (2 χ45 ml) and saturated aqueous NaCl (2 χ 45 mL). It was dried over Na ₂ SO ₄ and the solvent was evaporated under reduced pressure. Crystallization from ethanol (45 ml) was carried out. Yield: 3.103 g (74.8%), melting point: 94 - 95 ° C

IR (KBr, cm-1): 3426,3088, 1731, 1702, 1598, 1503,1394, 1345, 1241,1091, 898, 811,742,648

1 H-NMR (300 MHz, CDCl ₃ ): δ (ppm) = 1.25 (t, 3H, J = 7.2 Hz, -COOCH ₂ CH ₃ ); 1.70 -1.80 (m, 4H, -CH ₂ CH ₂ CH ₂ CH ₂ COOCH ₂ -); 2.344-2.43 (m, 2H,

CH ₂ CH ₂ CH ₂ CH ₂ COOCH ₂ -); 3.96 - 4.07 (m, 2H, 2CH ₂ CH ₂ CH ₂ CH ₂ COOCH ₂ -); 4.13 (q, 2H, J = 7.2 Hz, -COOCH ₂ CH ₃ ); 4.70 (s, 2H, -OCH ₂ -); 7.08 (d, 1H, J =

8.7 Hz, BO-H); 7.86 (d, 1H, J = 2.6 Hz, BO-H); 7.96 (dd, 1H, J1 = 8.9 Hz, J2 = 2.5 Hz, BO-H)

M (calculated) = 322; found (FAB) = 323 (MH +)

Elemental analysis for Ci5HigN ₂ O ₆ : calculated measured% C 55.90 55.74% H 5.63 5.63% N 8.69 8.67

Example 3:

Ethyl 5- (7-amino-3-oxo-2,3-dihydro-4 / 7-1,4-benzoxazin-4-yl) pentanoate (3)

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1.50 g (4.65 mmol) of compound 2 was dissolved in 310 ml of absolute ethanol. Purify with argon, add 0.130 g Pd / C (10%) and re-purge with argon. H ₂ is introduced into the reaction mixture. After two hours, the introduction was stopped and 0.065 g Pd / C (10%) was added. Then continue to introduce H ₂ into the suspension for another 40 minutes. The catalyst was filtered off by suction and the solvent was evaporated under reduced pressure. The product is light brown in color and oily. By trituration a compound 3 is ejected from the solvent mixture of ether and hexane.

Yield: 1.187 g (87.3%), melting point: 71 - 74 ° C

IR (KBr, cm-1): 3394, 3326, 3231, 2943, 1727,1658, 1515, 1423, 1312,1196, 1043,857, 801,651,586

H-NMR (300 MHz, CDCl ₃ ): δ (ppm) = 1.26 (t, 3H, J = 7.2 Hz, -COOCH ₂ CH ₃ );

1.66- 1.80 (m, 4H, -CH2CH2CH2CH2COOCH2-); 2.26-2.48 (m, 2H, CH2CH2CH2CH2COOCH2);

3.62 (s, 2H, BO-NH2); 3.83 - 3.98 (m, 2H, -CH ₂ CH ₂ CH ₂ CH ₂ COOCH ₂ -); 4.14 (q, 2H, J = 7.2 Hz, -COOCH ₂ CH ₃ ); 4.54 (s, 2H, -OCH ₂ -); 6.25 - 6.48 (m, 2H, BO-H);

6.66- 6.86 (m, 1H, BO-H)

M (calculated) = 292; found (FAB) = 293 (MH +)

Elemental analysis for C15H20N2O4: calculated measured % C 61.63 61.42 % H 6.90. 6.87 % N 9.58 9.71

Example 4:

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Ethyl 5- {7 - {(4-cyanobenzoyl) amino] -3-oxo-2,3-dihydro-4H-1,4-oxoxazin-4-ylpentanoate (4)

Dissolve 0.60 g (2.05 mmol) of compound 3 in 21 ml of pre-distilled chloroform and add 0.29 ml (2.05 mmol) of Ε ^ Ν. Then 0.34 g (2.05 mmol) of 4-cyanobenzoyl chloride is added while stirring. The reaction mixture was stirred at room temperature. After two days, 0.01 ml of Et ₃ N and 0.02 g of 4-cyanobenzoyl chloride are added. The reaction mixture was stirred at room temperature for another ten days. The solution was then washed successively with 1 M HCl (3 * 15 ml), water (3><15 ml) and saturated aqueous NaCl solution (3 × 15 ml). The organic phase was dried over Na ₂ SO 4 and the solvent was evaporated under reduced pressure. Crystallization from 30 ml of ethanol is carried out. Yield: 0.628 g (72.6%), melting point: 153 - 155 ° C

IR (KBr, cm-1): 3322, 2978, 2228, 1721, 1679, 1515, 1408, 1334, 1189, 1049, 861,762,540

H-NMR (300 MHz, CDCl ₃ ): δ (ppm) = 1.27 (t, 3H, J = 7.2 Hz, -COOCH2CH3); 1.68 -1.79 (m, 4H, -CH2CH2CH2CH2COOCH2-); 2.32 -2.43 (m, 2H, CH2CH2CH2CH2COOCH2); 3.93 - 4.10 (m, 2H, -CH2CH2CH2CH2COOCH2-); 4.14 (q, 2H, J = 7.2 Hz, -COOCH _? CH ·,): 4.62 (s, 2H, -OCH ₂ -); 6.98 (d, 1H, J =

8.7 Hz, BO-H); 7.29 -7.34 (m, 1H, BO-H); 7.38 (d, 1H, J = 2.3 Hz, BO-H); 7.81 (d, 2H, J = 8.3 Hz, Ar-H); 7.93 (s, 1H, -CONH-); 7.99 (d, 2H, J = 8.7 Hz, Ar-H)

M (calculated) = 421; found (FAB) = 422 (MH +)

Elemental analysis for C23H23N3O5: calculated measured % C 65.55 65.36 % H 5,50 5.70 % N 9.97 9.73

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Example 5:

5- {7 - {(4- (AMINOIMINOMETHYL) PHENYL) CARBOXAMIDO} -3-OXO-2,3-Dihydro-4H-1,4-BENZOXAZIN-4-yl} PENTANOIC ACETATE ACETATE ACETATE (5)

0.31 g (0.73 mmol) of compound 4 was added to 25 ml of absolute ethanol and the suspension cooled on ice. Gaseous HCl was introduced for 1 hour and then stirred at room temperature for 4 hours. Evaporate the solvent under reduced pressure and wash the residue with diethyl ether (anhydrous). It was then dissolved in 25 ml of absolute ethanol, 0.15 g (1.98 mmol) of ammonium acetate (anhydrous) was added and stirring was started at room temperature. After two days, the precipitate was removed, which was filtered off by suction and washed with absolute ethanol. The mother liquor was concentrated and 0.05 g of ammonium acetate was added. The precipitate again falls out and is filtered off by suction the next day and washed with absolute ethanol. Yield: 0.235 g (64.9%), melting point: 155 - 165 ° C

IR (KBr, cm-1): 3367, 2958, 1716, 1684, 1559, 1507, 1418, 1313, 1263, 1159, 1045,867,812,716,618 ¹ H-NMR (300 MHz, DMSO-d 6); δ (ppm) = 1.17 (t, 3H, J = 7.2 Hz, COOCH2CH3); 1.49 -1.64 (m, 4H, -CH2CH2CH2CH2COOCH2-); 1.79 (s, 3H, CH3COOH); 2.28-2.41 (m, 2H, -CH2CH2CH2COOCH2-); 3.85-4.00 (m, 2H, CH2CH2CH2CH2COO-); 4.04 (q, 2H, J = 7.2 Hz, -COOCH ₂ CH ₃ ); 4.64 (s, 2H, OCH2-); 7.21 (d, 1H, J = 9.0 Hz, BO-H); 7.42 -7.52 (m, 1H, BO-H); 7.56 (d, 1H,

J = 2.3 Hz, BO-H); 7.93 (d, 2H, J = 8.3 Hz, Ar-H); 8.05 (d, 2H, J = 8.3 Hz, Ar-H); 10.35 (s, 1H.-CONH-)

M (calculated) = 438; found (FAB) = 439 (MH +)

Elemental analysis for C23H26N4O5 * 1.35CH3COOH: calculated measured% C 59.41 59.45

-1818% Η 6.09% Ν 10.78

5.72

10.73

Reaction scheme for the preparation of compound 9:

A: BrCH ₂ COOEt, KF, dimethylformamide, B: BrCH ₂ COOEt, K ₂ CO ₃ , triethylbenzylammonium chloride, MeCN, C: H ₂ , Pd / C, EtOH, D: 4-cyanobenzoyl chloride, Et ₃ N, CHCI ₃ , E: 1. HCl (g), EtOH; 2. Ammonium acetate

Example 6:

Ethyl 2- (7-nitro-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-4-yl) acetate (6)

To a suspension of 1.00 g (5.15 mmol) of compound 1, 1.78 g (12.88 mmol) of K ₂ CO ₃ , 0.59 g (2.58 mmol) of benzyltriethylammonium chloride and 36 ml of acetonitrile were added to the honey

-1919 by stirring 0.69 ml (6.18 mmol) of α-bromoacetic acid ethyl ester and heating at 60 ° C in an oil bath for three days. The precipitate is filtered off. The solvent was evaporated under reduced pressure and the residue was dissolved in 72 ml of toluene. The organic phase was washed successively with 0.1 M HCl (2 * 15 ml), saturated aqueous NaHCO ₃ solution (2 x 15 ml) and saturated aqueous NaCl solution (1><15 ml). It is dried over Na ₂ SO ₄ and the solvent is evaporated.

Yield: 1,073 g (74.4%), melting point: 116 -119 ° C

IR (KBr, ^cm-1): 3095, 2999, 1733 1688, 1603 1521, 1376 1347, 1224, 1052, 890, 811.743, six hundred and first

¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1.32 (t, J = 7.2 Hz, 3H, CH ₂ CH ₃ ), 4.29 (q, J = 7.1 Hz, 2H. COOCH 2), 4.73 (s, 2H, OCH ₂ ), 4.79 (s, 2H, CH ₂ COO), 6.86 (d, J =

8.7 Hz, 1H, H _ar ), 7.91-7.98 (m, 2H, H _ar ) ppm.

M (calculated) = 280; found (FAB) = 281 (MH +)

Elemental analysis for Ci ₂ Hi ₂ N ₂ O ₆ : calculated measured % C 51.43 51.34 % H 4.29 4.28 % N 10.0 9.83

Example 7:

Ethyl 2- (7-amino-3-oxo-3,4-dihydro-2W-1,4-benzoxazin-4-yl) acetate (7)

0.70 g (2.50 mmol) of compound 6 was dissolved in 120 ml of absolute ethanol. After purging with argon, 0.08 g Pd / C (10%) was slowly added. H _{2 is} introduced into the suspension and the reaction begins after 20 minutes. The Pd / C was filtered off by suction and washed with ethanol. The solvent was evaporated under reduced pressure.

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Yield: 0.586 g (93.8%), melting point: 131 - 134 ° C

IR (KBr, ^cm-1): 3411, 3327, 3228, 2984, 1744, 1668, 1519, 1414, 1312, 1217, 1051.902, 797, 538th

¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1.29 (t, J = 7.2 Hz, 3H, CH ₂ CH ₃ ), 3.62 (s, 2H, NH ₂ ), 4.25 (q, J = 7.2 Hz, 2H, COOCH ₂ ), 4.62 (d, J = 7.9 Hz, 4H, OCH ₂ , CH ₂ COO), 6.32-6.39 (m, 2H , H _ar ), 6.56 (d, J = 8.7 Hz, 1H, H _ar ) ppm.

M (calculated) = 250; found (FAB) = 250 (MH +)

Elemental analysis for Οΐ2Η ₁₄ Ν ₂ Ο ₄ : calculated measured % C 57.60 57.62 % H 5.60 5.72 % N 11,20 11.02

Example 8:

ETHYL 2- {7 - [(3-CLANOPHENYL) CARBOXAMIDO] -3-OXO-3,4-Dihydro-2 / 7-1.4BENZOXAZIN-4-ILACETATE (8)

To 0.50 g (2.00 mmol) of compound 7 was added 0.28 ml (2.00 mmol) of Et ₃ N, 20 ml of pre-distilled chloroform and finally 0.33 g (2.00 mmol) of 3-cyanobenzoyl chloride. The reaction mixture was stirred at room temperature. After 24 hours, the filtered compound was removed. 0.52 g of the compound is obtained. The remaining solution was washed successively with 1 M HCl (2 x 20 ml), water (2 x 20 ml) and saturated aqueous NaCl solution (1 x 20 ml). The organic phase was dried over Na ₂ SO ₄ and the solvent was evaporated. 0.12 g of the compound is obtained.

Yield: 0.641 g (84.6%), melting point: 207 - 211 ° C

-2121

IR (KBr, cm ¹ ): 3295, 2230, 1732, 1692, 1657, 1517, 1403, 1340, 1217, 1057, 875, 681.

¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1.31 (t, J = 7.2 Hz, 3H, CH? CHA 4.28 (q, J = 7.2 Hz, 2H, COOCH ₂ ) , 4.70 (d, J = 11.3 Hz, 4H, OCH ₂ , CH ₂ COO), 6.77 (d, J =

8.7 Hz, 1H, H _ar ), 7.28 (s, 1H, NH), 7.43 (d, J = 2.3 Hz, 1H, H _ar ), 7.66 (m, 1H, H) »), 7.86 (m, 2H, H _ar ), 8.12-8.19 (m, 2H, H _ar ) ppm.

M (calculated) = 379; found (El) = 379

Elemental analysis for C20H17N3O5: calculated measured % C 63.32 63.11 % H 4,48 4.61 % N 11.08 11.04

Example 9;

ACETATE OF ETHYL ESTER 2- {7 [(4- (AMINOIMINOMETHYL)

FENlL) CARBOXAMIDO] -3-OXO-3,4-DIHYDRO-2 / - / - 1,4-BENZOXAZ1N-4-IL} ACETIC ACIDS (9)

0.25 g (0.66 mmol) of compound 8 was dissolved in 70 ml of absolute ethanol and the solution cooled on ice. Introduce gaseous HCl for 45 minutes and then stir for a further 6 hours at room temperature. The solvent was evaporated and the residue was washed with diethyl ether. It was dissolved in 20 ml of absolute ethanol, 0.15 g (1.98 mmol) of ammonium acetate was added and stirring was started at room temperature. After four days, the precipitate disappears, which is filtered off and washed with absolute ethanol. Yield: 0.137 g (45.5%), melting point: 200 - 204 ° C

IR (KBr, ^cm-1): 2987, 1750, 1694, 1612, 1559, 1515, 1415, 1284, 1222, 1153, 1050, 861, 707th

-2222 ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ = 1.22 (t, J = 7.2 Hz, 3H, CH ₂ CH ₃ ), 4.17 (q, J = 7.1 Hz, 2H, COOCH ₂ ), 4.73 (s, 4H, OCH ₂ , CH ₂ COO), 7.08 (d, J = 9.0 Hz, 1H, H _ar ), 7.45 (m, 1H , H _ar ), 7.58 (d, J = 2.3 Hz, 1H, H _ar ), 7.93 (d, J = 8.7 Hz, 2H, H _ar ), 8.08 (d, J = 8.3 Hz, 2H, H _ar ), 10.43 (s, 1 H, NH) ppm.

M (calculated) = 396; found (FAB) = 397

Reaction Scheme for the Preparation of Compound 16:

-2323

A: EtOOCC (CH ₃ ) BrCOOEt, KF, dimethylformamide; B: MeJ, KF, dimethylformamide; C: NaOH, dioxane; D: GlyOEt, EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide

-2424 hydrochloride), HOBT (1-hydroxybenzotriazole), N-methylmorpholine, DMF; E: H ₂ , Pd / C, EtOH; F: 4-cyanobenzoyl chloride, Et ₃ N, CHCl ₃ , G: 1. HCl (g), EtOH; 2. Ammonium acetate, EtOH; H: NH ₂ OH, EtOH; l: EtOCOCI, pyridine; J: NaOH, EtOH; K: H ₂ , Pd / C, CH ₃ COOH, dimethylformamide

Example 10;

Ethyl 2-methyl-7-nitro-3-oxo-3,4-dihydro-2H-1,4-benzoxazine-2carboxylate (10)

To a suspension of 5.81 g (100.0 mmol) of KF in 25 ml of anhydrous DMF was added 10.12 g (40.0 mmol) of diethyl 2-bromo-2-methylmalonate while stirring. After stirring at room temperature for 30 min, 6.17 g (40.0 mmol) of 2-amino-5-nitrophenol were added and allowed to stir overnight in an oil bath at 60 ° C. The reaction mixture was cooled to room temperature and gradually poured into a mixture of 60 g of ice and 40 ml of water. The precipitated precipitate was filtered off by suction, washed with cold water and the product (compound 10) was recrystallized from anhydrous ethanol (ca. 90 ml). Needle crystals of pink color are obtained. Efficiency; 9.30 g (83%), melting point: 191-193 ° C

IR (KBr, cm ' ¹ ): 3088, 1749, 1697, 1605, 1540, 1508, 1340, 1237, 1123, 1012,

826, 744 ¹ H-NMR (300 MHz. DMSO): δ (ppm) = 1.07 (t, 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.75 (s, 3H, 2- CH ₃ ), 4.12 (m, 2H, CH ₂ ), 7.10 (d, 1H, J = 8.67 Hz, Ar-Hs), 7.84 (d, 1 H,

J = 2.64 Hz, Ar-Hs), 7.95 (dd, 1H, J = 9.04, 2.63 Hz, Ar-H ₆ ), 11.60 (wid s, 1H, NH)

Mr (calculated) = 280; found (FAB) = 281 (MH ⁺ )

-2525

Example 11:

Ethyl 2,4-dimethyl-7-nitro-3-oxo-3,4-dihydro-2H-1,4-benzoxazine-2carboxylate (11)

To a suspension of 2.91 g (20.0 mmol) of KF in 40 ml of DMF was added 5.60 g (20.0 mmol) of compound 10 and 1.32 ml (21.0 mmol) of methyl iodide. The reaction mixture was stirred for 16 hours at 40 ° C and then poured onto 80 g of ice. A white precipitate precipitates, which is filtered off by suction, washed with water and recrystallized from isopropanol. Yield: 5.68 g (96%), melting point: 118-120 ° C. ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.05 (t, 3H, J = 7.15 Hz, CH ₂ CH ₃ ), 1.78 (s, 3H, 2-CH ₃ ), 3.40 (s, 3H, N-CH ₃ ), 4.10 (m, 2H, CH ₂ ); 7.43 (d, 1H, J = 8.67 Hz, Ar-H ₅ ), 7.87 (d, 1H, J = 2.26 Hz, Ar-H ₈ ), 8.02 (dd, 1H. J = 9.04, 2.63 Hz, Ar-H ₆ )

Mr (calculated) = 294; found (FAB) = 295 (MH ⁺ )

Example 12:

2,4-Dimethyl-7-nitro-3-oxo-3,4-dihydro-2 / 7-1,4-benzoxazine-2-carboxylic acid (12)

5.88 g (20.0 mmol) of compound 11 was dissolved in 70 ml of dioxane and 15.0 ml (30 mmol) of 2M NaOH was added while stirring. After 24 hours, the dioxane was evaporated from the reaction mixture under reduced pressure and the residue was dissolved in 60 ml of water. The resulting solution was extracted with ethyl acetate (2 x 50 mL), acidified with 4M HCl to pH 2, and extracted with ether (2x50 mL). The ether phase is dried over Na ₂ SO4, the solvent is evaporated off under reduced pressure and the solid residue is dried on a diaphragm pump. Yield: 5.00 g (94%), melting point: 158Ί60 ° C

-2626

IR (KBr, ^cm-1): 3540, 3381, 1760, 1678, 1599, 1527, 1341 1250 1114 1026 871 745 ¹ H NMR (DMSO, 300 MHz) *: δ (ppm) = 1, 76 (s, 3H, 2-CH ₃ ), 3.39 (s, 3H, NCH ₃ ), 7.41 (d, 1H, J = 9.05 Hz, Ar-H ₅ ), 7.84 (d , 1H, J = 2.64 Hz, Ar-H ₈ ), 7,988.02 (dd, 1 H, J = 9.04, 2.64 Hz, Ar-H _e ) * signal for COOH not visible, signal for H ₂ O is shifted to 3.57 ppm

Mr (calculated) = 266; found (El) = 266

HRMS: Mr for ΟιιΗ ₁₀ Ν ₂ Ο ₆ (calculated) = 266.0539; found (El) = 266.0549

Example 13:

ETHYL {{(2,4-DIMETHYL-7-NITRO-3-OXO-3,4-DIHYDRO-2A7-1,4-BENZOXAZIN-2IL) CARBONYLAMINOJACETATE (13)

0.96 g (3.6 mmoi) of acid 12 and 0.54 g (3.8 mmol) of the glycine ethyl ester of hydrochloride are dissolved in 20 ml of anhydrous DMF, cooled to -10 ° C and stirred for 10 minutes Add HOBT (0 , 66 g; 4.3 mmol) followed by NMM (1.22 ml; 11.0 mmol) and finally EDC (0.90 g; 4.6 mmol). The reaction mixture was stirred for 24 hours, gradually increasing the T from -10 ° C to room temperature. The DMF was evaporated under reduced pressure and the residue was taken up in ethyl acetate (40 ml). Extract with 10% citric acid aqueous solution (2 x 30 ml), saturated NaHCO ₃ solution (2 x 30 ml) and saturated NaCl solution (30 ml). The organic phase was dried with Na ₂ SO4, filtered and the solvent was evaporated under reduced pressure. A pure yellowish product is obtained which is dried on a diaphragm pump. Yield: 0.92 g (73%), mp 147-150 ° C

IR (KBr, cm ' ¹ ): 3340, 2993, 1746, 1697, 1602, 1531, 1342, 1207, 1128, 1070, 1027, 898, 816, 746, 589

-2727 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.07 (t, 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.78 (s, 3H, 2-CH ₃ ), 3.37 (s, 3H, N-CH ₃ ), 3.72 (d, 2H, J = 6.02 Hz, NH-CH ₂ -), 3.96 (q, 2H, J = 7. 16 Hz, -CH ₂ CH ₃ ), 7.39 (d, 1H, J = 8.66 Hz, Ar-H ₅ ), 7.98-8.02 (m, 2H, Ar-H ₆ , H _e ), 8.73 (t, 1 H, J = 6.02 Hz, NH)

Mr (calculated) - 351; found (FAB) ~ 352 (MH ⁺ )

Elemental analysis for C15H17N3O7'0.25 H ₂ O: calculated% C 50.63% H 4.96% N 11.81 measured

50.51

5.10

11.60

Example 14:

ETHYL {{(7-AMINO-2,4-DIMETHYL-3-OXO-3,4-DIHYDRO-2H-1,4-BENZOXAZIN-2IL) CARBONYL} AMINO} ACETATE (14)

755 mg (2.15 mmol) of Compound 13 was dissolved in 120 ml of anhydrous ethanol, aspirated with argon, 100 mg Pd / C added, aspirated again with argon and hydrogen stirred for 1 hour. The reaction mixture was purged with argon and the catalyst was filtered off by suction and the solvent was evaporated off under reduced pressure. Drying gives a pure product. Yield: 675 mg (97.7%), melting point: 146-148 ° C

IR (KBr, cm ' ¹ ): 3380, 2989, 1743, 1681, 1639, 1518, 1394, 1312, 1187, 1018, 836, 615 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.13 (t, 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.66 (s, 3H, 2-CH ₃ ), 3.21 (s, 3H, N-CH ₃ ), 3.66- 3.74 (m, 2H, NH-CH ₂ -), 4.02 (m, 2H, CH ₂ CH ₃ ), 5.56 (s, 2H, NH ₂ ), 6.31-6.35 (dd , 1H, J = 8.29, 2.27 Hz, Ar-H ₆ ), 6.39

-2828 (d, 1H, J = 2.26 Hz, Ar-H ₈ ), 6.82 (d, 1H, J = 8.66 Hz, Ar-H ₅ ), 8.37 (t, 1H, J = 6.02 Hz, NH)

Mr (calculated) = 321; found (El) = 321

Example 15:

ETYL2 - {{(7 - {(4-CYANOBENZOIL) AMINO} -2,4-DIMETHYL-3-OXO-3,4-DIHYDRO2H-1,4-BENZOXAZIN-2-yl) CARBONYL} AMINO} ACETATE (15)

645 mg (2.0 mmol) of amine 14 was dissolved in 25 ml of pre-distilled dichloromethane, 0.29 ml (2.1 mmol) of Et ₃ N was added and cooled to -10 ° C. 355 mg (2.1 mmol) of 4-cyanobenzoyl chloride was added and allowed to stir overnight, with T gradually increasing to room temperature. After 16 hours, the solvent was evaporated, the residue was dissolved in 40 ml of ethyl acetate and extracted with water (30 ml), 10% aqueous citric acid solution (2 x 30 ml), saturated NaHCO ₃ solution (2 x 30 ml) and saturated NaCl solution ( 30 ml). The organic phase was dried with Na ₂ SO4, filtered and the solvent was evaporated under reduced pressure. The residue was taken up in ether (15 ml) and the precipitate was filtered off by suction. Yield: 681 mg (76%); melting point: 171174 ° C.

IR (KBr, cm · ¹ ): 3376, 2986, 2228, 1734, 1697, 1514, 1431, 1389, 1221, 1151, 1018, 865, 761,634 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1 , 11 (t, 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.73 (s, 3H, 2-CH ₃ ), 3.30 (s, 3H, N-CH 3), 3.62 3.80 (m, 2H, NH-CH ₂ -), 4.01 (m, 2H, CH ₂ CH ₃ ), 7.15 (d, 1H, J = 9.04 Hz, Ar-H ₅ ) , 7.39-7.42 (dd, 1H, J = 8.66, 2.26 Hz, Ar-He), 7.74 (d, 1H, J = 2.26 Hz, Ar-H ₈ ), 8.02 (d, 2H, J = 8.29 Hz, Ar-2H), 8.10 (d, 2H, J = 8.67 Hz, Ar-2H), 8.54 (t, 1H, J = 6.03 Hz, CO-NH-CH ₂ ), 10.51 (s, 1H, CONH-Ar)

-2929

Mr (calculated) = 450; found (FAB) = 451, (MH ⁺ )

Elemental analysis for C23H22N4O6: calculated measured % C 61.33 61.07 % H 4.92 4,85 % N 12,44 12,45

Example 16:

ETHYL 2 - {{(7 - {(4- {AMINO (IMINO) METHYL} BENZOIL) AMINO} -2,4-DIMETHYL-3-OKSO3,4-DIHYDRO-2H-1,4-BENZOXAZIN-2-yl) CARBON ILYAMINE OJACETAT

600 mg (1.33 mmol) of compound 15 was dissolved in 30 ml of anhydrous ethanol and HCI (g) was added to saturation (45 minutes) while stirring on an ice bath (0 ° C). The mixture was stirred at room temperature for 24 hours, then the solvent was evaporated off under reduced pressure and the residue was suspended in 20 ml of pre-distilled diethyl ether. The compound was filtered off by suction, dried on a membrane pump, 310 mg (4.0 mmol) of purified ammonium acetate were added and the mixture was dissolved in 20 m anhydrous ethanol. Stirred for 24 hours at room temperature. A few approx. 5 ml of diethyl ether and cool in the freezer. After 2 hours, the ammonium chloride was filtered off and the mother liquor was concentrated to approx. 10 ml and leave it in the freezer overnight. Compound 16 was removed, which was filtered off by suction and washed with a few ml of ethanol and diethyl ether. Yield: 330 mg (47%), melting point: 146-149 ° C

IR (KBr, cm ' ¹ ): 3318, 2988, 1688, 1512, 1384, 1260, 1153, 1018,863, 649 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.11 (t, 3H , J = 7.16 Hz, CH ₂ CH ₃ ), 1.73 (s, 3H, 2-CH ₃ ), 1.76 (s, 3H, CH ₃ COOH), 3.30 (s, 3H, N-CH ₃ ), 3.63-3.79 (m, 2H, NHCH ₂ -), 4.00 (q, 2H, J = 7.16 Hz, -CH ₂ CH ₃ ), 7.15 (d, 1H. J = 9.04 Hz, Ar-Hs), 7,417,45 (dd, 1H, J = 8.66, 2.26 Hz, Ar-H _6), 7.76 (d, 1 H, J = 1, 89 Hz, Ar-Hs), 7.94 (d,

-3030

2Η, J = 8.29 Hz, Ar-2H), 8.11 (d, 2H, J = 8.29 Hz, Ar-2H), 8.54 (wid s, 1H, CONH-CH ₂ ), 10.48 (s, 1H, CO-NH-Ar) * protons of amidine group and signal for H ₂ O not visible, baseline offset

Mr (calculated) = 467 (527 as acetate); found (FAB) = 468 (MH ⁺ )

Elemental analysis for C23H25N5O6 '1.2 CH ₃ COOH 1 H ₂ O:

calculated measured% C 54.72 55.06% H 5.75 6.03% N 12.56 12.17

Example 17:

ETHYL 2 - {{(74 (4- {AMINO (HYDROXYIMINO) METHYL} BENZOIL) AMINO} -2,4-DIMETHYL-OXO-3,4-Dihydro-2H-1,4-BENZOXAZIN-2-yl) CARBONYL}

AMINO} ACETATE (17)

Hydroxylammonium chloride (215 mg; 3.0 mmol) was dissolved in 20 ml of anhydrous ethanol, 0.42 ml Et ₃ N (3.0 mmol) was added and stirred for 20 minutes. To the mixture was added 450 mg (1.0 mmol) of compound 15 and 10 ml of ethanol and heated at 50 ° C for 3 hours and then at room temperature overnight. The reaction mixture was cooled in the freezer and the precipitate was filtered off.

Yield: 340 mg (70.4%), melting point: 150-153 ° C

IR (KBr, cm ' ¹ ): 3346, 2986, 2936, 1732, 1697, 1608, 1516, 1430, 1389, 1317, 1253, 1161, 1088, 1015,928,859,814,705 ¹ H NMR (DMSO, 300 MHz): δ ( ppm) = 1.11 (t, 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.73 (s, 3H, 2-CH ₃ ), 3.32 (s, 3H, N-CH ₃ ), 3.63-3.80 (m, 2H, NH-CH 2 -), 4.00 (q, 2H, J = 7.16 Hz, -CH ₂ CH ₃ ), 5.93 (s, 2H. NH ₂ ), 7.13 (d, 1H, J = 8.67 Hz, Ar-H _s ), 7.40-7.44

-3131 (dd, 1H, J = 8.67, 2.26 Hz, Ar-H ₆ ), 7.76 (d, 1H, J ~ 2.26 Hz, Ar-H _B ), 7.83 (d , 2H, J = 8.29 Hz, Ar-2H), 7.95 (d, 2H, J = 8.66 Hz, Ar-2H), 8.53 (t, 1H, J = 6.03 Hz. CO-NHCH ₂ ), 9.83 (s, 1H, OH), 10.29 (s, 1H, CO-NH-Ar)

Mr (calculated) = 483; found (FAB) = 484 (MH ⁺ )

Elemental analysis for C23H25N5O7 0.5 H ₂ O: calculated% C 56.09% H 5.32% N 14.22 measured

56.18

5.37

14,31

Example 18:

ETYL2 - {{(2,4-DIMETHYL-3-OXO-7 - {(4- (5-OXO-4,5-Dihydro-1,2,4-OXADIAZOL · 3-yl) BENZOIL) AMINO} -3 , 4-DIHYDRO-2H-1,4-BENZOXAZIN-2-yl) CARBONYL} AMINOJACETATE (18)

300 mg (0.62 mmol) of compound 17 was dissolved in 12 ml of pre-distilled pyridine, purged with argon and cooled to -15 ° C. Add 0.062 ml (0.66 mmol) of ethyl chloroformate and stir for 45 minutes in an ice bath. The temperature was then raised to room temperature and stirred for 30 minutes, the mixture was bubbled with argon and heated for 6 hours at boiling point (ca. 120 ° C). The pyridine was then evaporated under reduced pressure and 30 ml of methanol was added to the residue and the resulting product was filtered off. Yield: 180 mg (57%), melting point: 221-225 ° C

IR (KBr, cm ' ¹ ): 3366, 2987, 1760, 1693, 1513, 1430, 1250, 1150, 1018, 862, 758 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.11 (t , 3H, J = 7.16 Hz, CH ₂ CH ₃ ), 1.73 (s, 3H, 2-CH ₃ ), 3.34 (s, 3H, N-CH ₃ ), 3.62-3.80 (m, 2H, NH-CH ₂ -), 4.00 (q, 2H, J = 7.16 Hz, -CH ₂ CH ₃ ), 7.15 (d, 1H, J = 9.05 Hz, Ar -H ₅ ), 7.40-7.44 (dd, 1H, J = 8.66, 2.26 Hz, Ar-H _s ), 7.76 (d, 1H, J = 2.26 Hz, Ar -H ₈ ), 7.96 (d, 2H, J = 8.66 Hz, Ar-2H),

-3232

8.12 (d, 2H, J = 8.67 Hz, Ar-2H), 8.54 (t, 1H, J = 6.02 Hz, CO-NH-CH ₂ ), 10.45 (s, 1H , CO-NH-Ar), 13.11 (dec. S, 1H, NH-form)

Mr (calculated) = 509; found (FAB) = 510 (MH ⁺ )

Elemental analysis for C24H ₂₃ N ₅ O ₈ 0.25 H ₂ O: calculated% C 56.08% H 4.61% N 13.63 measured

55.98

4,42

13,47

Example 19:

2 - {{(2,4-Dimethyl-3-oxo-7 - {(4- (5-oxo-4,5-dihydro-1,2,4-oxadiazol-3yl) benzoyl) AMINO} -3,4 -Dihydro-2H-1,4-benzoxazazin-2-yl) carbonyl} aminoacetic acid (19)

150 mg (0.29 mmol) of compound 18 was suspended in 10 ml of ethanol, 0.3 ml (0.6 mmol) of 2M NaOH was added and stirred for 4 hours at room temperature. To the reaction mixture was added 1 ml of acetic acid and left in the freezer overnight. The precipitate was filtered off by suction, washed with chilled ethanol and dried at a diaphragm pump.

Yield: 140 mg (99%), melting point: 206-209 ° C

IR (KBr, cm ^-1 ): 3447, 1779, 1680, 1552, 1513, 1436, 1290, 1205, 1032, 946, 816, 756, 670 ¹ H NMR (DMSO, 300 MHz) *: δ (ppm) = 1.72 (s, 3H, 2-CH ₃ ), 3.30 (s, 3H, N-CH ₃ ), 3.52-3.75 (m, 2H, NH-CH ₂ -), 7.13 (d, 1H, J = 9.05 Hz, Ar-H ₅ ), 7.40-7.44 (dd, 1H, J = 8.66, 2.26 Hz, Ar-H ₆ ), 7.72 (d, 1H, J = 2.26 Hz, Ar-H ₈ ), 7.96 (d, 2H, J = 8.66 Hz, Ar-2H), 8.11 (d, 2H, J = 8. 29 Hz, Ar-2H), 8.38 (t, 1H, J = 5.98 Hz, CO-NH-CH ₂ ), 10.42 (s, 1H, CO-NH-Ar), 12.66 ( s, 1H, NH-turn) * COOH signal not visible, water distortion signal

Mr (calculated) = 481; found (FAB) = 482 (MH ⁺ )

-3333

Elemental analysis for C ₂₂ Hi9N ₅ O ₈ '1.75 H ₂ 0: calculated% C 51.51% H 4.42% N 13.65 measured

51.55

4.38

13,34

Example 20:

2 - {{(7 - {(4- {AMINO (1MINO) METHYL} BENZOIL) AMINO} -2,4-DIMETHYL-3-OXO-3,4-Dihydro-2H-1,4-BENZOXAZIN-2-yl) CARBONYL } AMINO} ACETIC ACID (20)

Dissolve 110 mg (0.23 mmol) of compound 19 in a mixture of 10 ml of DMF and 5 ml of acetic acid, purge with argon, add 30 mg of Pd / C, re-purge with argon and stir with hydrogen for 4 hours. The reaction mixture was purged with argon and the catalyst was filtered off by suction. Add 40 ml of diethyl ether to the mother liquor and filter out the precipitate. Yield: 82 mg (72%), melting point: 242-245 ° C

IR (KBr, cm ' ¹ ): 3338.1659, 1514, 1389, 1314, 1150, 1032, 863, 703 ¹ H NMR (DMSO + D ₂ O, 300 MHz) *: δ (ppm) = 1.74 ( s, 3H, 2-CH ₃ ), 7.16 (d, 1H, J = 9.05 Hz, Ar-Hs), 7.32-7.37 (dd, 1H, J = 9.04, 1. 88 Hz, Ar-H ₆ ), 7.49 (t, 1H, J = 4.90 Hz, CO-NH-CH 2), 7.68 (d, 2H, J = 8.67 Hz, Ar-2H) , 7.86 (d, 1H, J = 1.89 Hz, ArH ₈ ), 8.01 (d, 2H, J = 8.66 Hz, Ar-2H), 10.42 (s, 1H, CO -NH-Ar) * signals between 3.1 and 3.9 ppm (N-CH ₃ , NH-CH ₂ -) are not visible because they are covered by the water signal (D ₂ O as solvent). Also, no signals for the basic amidine and acidic COOH group are visible.

Elemental analysis for C ₂₁ H ₂₁ N ₅ O ₆ CH ₃ COOH 2.25 H ₂ O:

calculated measured% C% H% N

51.18

5.51

12,97

50.88

5.63

13,27

-3434

Mr (calculated) = 439; found (FAB) = 440 (MH ⁺ )

Reaction Scheme for Preparation of Compound 25;

A; 1. EtOCOCI, Et ₃ N, THF; 2. TMSCHN ₂ (trimethylsilyldiazomethane), MeCN; B: mercury benzoate, Et ₃ N, MeOH, ultrasound; C: H ₂ , Pd / C, MeOH; D: 4cyanobenzoyl chloride, CH ₂ Cl ₂ , E: 1. HCl (g), MeOH; 2. Ammonium acetate, MeOH

Example 21;

2- (DIAZOACETYL) -2,4-DIMETHYL-7-NITRO-3-2H-1,4-BENZOXAZIN-3 (4H) -ONE (21)

Dissolve 1.6 g (6.0 mmol) of acid 12 in 30 ml of freshly pre-distilled THF, add 0.86 ml (6.2 mmol) of Et ₃ N, blend with argon and cool to -15 ° C.

-3535

A solution of 0.59 ml (6.2 mmol) of ethyl chloroformate in 5 ml of THF was added and stirred for 30 minutes at -5 ° C. The precipitated triethylammonium chloride is filtered off. 20 ml of acetonitrile was added to the filtrate, aspirated with argon and cooled to 0 ° C. 6.0 ml (12.0 mmol) of trimethycyldyldiazomethane (2.0 M solution in hexane from Aidrich® manufacturer) were added to the mixture and stirred at 4 ° C for 24 hours. Then 80 ml of diethyl ether are added and extracted with 10% aqueous citric acid solution (2 x 30 ml), saturated NaHCOs solution (2 x 30 ml) and saturated NaCl solution (30 ml). The organic phase was dried with Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. The crude product is used in the next step.

Yield: 1.59 g (91.7%), melting point: 128-130 ° C

IR (KBr, cm ' ¹ ): 3452, 3085, 2126, 1691, 1636, 1600, 1525, 1341, 1108, 1022, 876, 744, 531 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1 , 73 (s, 3H, 2-CH ₃ ), 3.38 (s, 3H, N-CH ₃ ), 6.48 (s, 1H, CH-Nz), 7.40 (d, 1H, J = 8.66 Hz, Ar-H ₅ ), 7.89-8.02 (m, 2H, ArH ₈ , H ₆ )

Mr (calculated) = 290; found (El) = 290

HRMS: Mr for Ci ₂ H ₁₀ N ₄ O ₅ (calculated) = 290.0651; found (El) = 290.0662

Example 22:

METHYL 2- (2,4-DIMETHYL-7-NITRO-3-OXO-3,4-DIHYDRO-2H-1,4-BENZOXAZIN2-IL) ACETATE (22)

1.45 g (5.0 mmol) of diazoketone 21 were suspended in 50 ml of anhydrous methanol. The mixture was placed in an ultrasonic bath and a solution of 0.23 g (1.0 mmol) of freshly prepared silver benzoate in 2.8 ml (20 mmol) Et ₃ N was gradually added. After 30 minutes the methanol was evaporated under reduced pressure, the residue was dissolved in 60 ml.

-3636 ethyl acetate and extracted with 10% aqueous citric acid solution (2 x 20 ml), saturated NaHCO ₃ solution (2 x 20 ml) and saturated NaCl solution (30 ml). The organic phase was dried with Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. The product was purified by column chromatography. As the mobile phase, a mixture of ethyl acetate and hexane in a ratio of 3: 2 was used to give an oily product.

Yield: 1.23 g (83.7%)

IR (NaCl. Cm ' ¹ ): 2953.1741,1693, 1600, 1524, 1322,1209,1074, 881,744 ¹ H NMR (CDCl ³ , 300 MHz): δ (ppm) = 1.54 (s, 3H, 2-CH ₃ ), 2.86 (d, 1H, J = 16.58 Hz, CH ₂ ), 3.34 (d, 1H, J = 16.58 Hz, CH ₂ ), 3.46 (s, 3H, N -CH ₃ ), 3.68 (s,

3H, O-CH ₃ ), 7.03 (d, 1H, J = 9.04 Hz, Ar-H ₅ ), 7.81 (d, 1H, J = 2.64 Hz, Ar-H ₈ ), 7.94-7.98 (dd, 1H, J = 9.04, 2.63 Hz, Ar-Ηβ)

Mr (calculated) = 294; found (El) = 294

HRMS: Mark for Ci3Hi ₄ N ₂ O ₆ (calculated) = 294.0852; found (El) = 294.0862

Example 23:

Methyl 2- (7-amino-2,4-dimethyl-3-oxo-3,4-dihydro-2H-1,4-benzoxazin2-yl) acetate (23)

Dissolve 382 mg (1.3 mmol) of compound 22 in 40 ml of anhydrous methanol, argon, add 40 mg Pd / C, argon again and hydrogen for 2 hours under stirring. The reaction mixture was purged with argon and the catalyst was filtered off by suction and the solvent was evaporated off under reduced pressure. The crude oily product obtained after drying is used in the next step. Yield: 324 mg (94.6%)

IR (NaCl, cm ' ¹ ): 3361,2951,1740, 1665, 1518,1398, 1312,1192,1146,1011

-3737

Mr (calculated) = 264; found (FAB) = 265 (MH ⁺ )

Example 24:

Methyl 2- {7 - {(4-cyanobenzoyl) amino] -2,4-dimethyl-3-oxo-3,4-dihydro2H-1,4-benzoxazin-2-yl} acetate (24)

Dissolve 320 mg (1.21 mmol) of amine 23 in 10 ml of pre-distilled dichloromethane, add 0.18 ml (1.30 mmol) of Et ₃ N and cool to -10 ° C. 203 mg (1.20 mmol) of 4-cyanobenzoyl chloride were added and allowed to stir overnight, with T gradually increasing to room temperature. After 18 hours, the solvent was evaporated, the residue was dissolved in 30 ml of ethyl acetate and extracted with 10% aqueous citric acid solution (2 x 20 ml), saturated NaHCO ₃ solution (2 x 20 mi) and saturated NaCl solution (20 ml). The organic phase was dried with Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. The residue was taken up in ether (10 mi) and the precipitate was filtered off by suction.

Yield: 352 mg (74%), melting point: 138-142 ° C

IR (KBr, cm ' ¹ ): 3372, 2953, 2228, 1732, 1673, 1517, 1394, 1215, 1153, 1000, 866, 635 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1.41 (s, 3H, 2-CH ₃ ), 2.83 (d, 1H, J = 16.21 Hz, CH ₂ ), 3.15 (d, 1H, J = 16.20 Hz, CH ₂ ), 3 , 29 (s, 3H, N-CH ₃ ), 3.57 (s, 3H, O-CH ₃ ), 7.14 (d, 1H, J = 8.67 Hz, Ar-H ₅ ), 7. 41-7.45 (dd, 1H, J = 8.67, 2.26 Hz, Ar-He), 7.49 (d, 1H, J = 2.26 Hz, Ar-He), 8.02 ( d, 2H, J = 8.67 Hz, Ar-2H), 8.09 (d, 2H, J = 8.67 Hz, Ar-2H), 10.45 (s, 1 H, NH)

Mr (calculated) = 393; found (FAB) = 394 (MH ⁺ )

Elemental analysis for C ₂ andH ₁ 9N ₃ O ₅ · 0,5 H ₂ O: calculated measured% C 62.68 62.63

-3838% Η 5.01% Ν 10.44

4.63

10.55

Example 25:

METHYL 2- {7 - {(4- {AMINO (IMINO) METHYL} BENZOIL) AMINO} -2,4-DIMETHYL-3-OKSO3,4-Dihydro-2H-1,4-BENZOXAZIN-2-yl} ACETATE ( 25)

330 mg (0.84 mmol) of compound 24 was dissolved in 20 ml of anhydrous methanol and HCI (g) was added to saturation (40 minutes) while stirring on an ice bath (0 ° C). The mixture was stirred at room temperature for 17 hours, then the solvent was evaporated off under reduced pressure and the residue was poured into 20 ml of pre-distilled diethyl ether. The ether was carefully decanted, the oily residue was dissolved in methanol and the process repeated. 300 mg (3.90 mmol) of purified ammonium acetate was added and the mixture was dissolved in 10 ml of anhydrous methanol. The mixture was stirred at room temperature for 24 hours and then cooled in a freezer. The precipitate was filtered off by suction.

Yield: 47 mg (12%), melting point: 180-184 ° C

IR (KBr, cm ' ¹ ): 3391, 2946, 1713, 1670, 1606, 1516, 1413, 1314, 1238, 1149, 1009, 860, 705 ¹ H NMR (DMSO, 300 MHz): δ (ppm) = 1 , 41 (s, 3H, 2-CH ₃ ), 1.80 (s, 3H, CH ₃ COOH), 2.83 (d, 1H, J = 16.20 Hz, CH ₂ ), 3.15 (d , 1H, J = 16.20 Hz, CH ₂ ), 3.30 (s, 3H, N-CH ₃ ), 3.57 (s, 3H, O-CH ₃ ), 7.14 (d, 1H. J = 9.04 Hz, Ar-H ₅ ), 7,447.48 (dd, 1H, J = 8.67, 2.26 Hz, Ar-H _e ), 7.51 (d, 1 H, J = 1 , 89 Hz, Ar-H ₈ ), 7.93 (d, 2H, J = 8.29 Hz, Ar-2H), 8.10 (d, 2H, J = 8.28 Hz, Ar-2H), 10.43 (bs, 1H, NHCO) * protons of amidine group and signal for Η ₂ Ο not visible, baseline offset

Mr (calculated) = 410 (470 as acetate); found (FAB) = 411 (M

-3939

Reaction scheme for the preparation of compound 28:

A: H ₂ , Pd / C, MeOH; B: 4-cyanobenzoyl chloride, CHCl ₃ , C: 1. HCl (g), EtOH; 2. Ammonium acetate, EtOH

Example 26:

ETYL7-AMINO-3,4-DIHYDRO-2-METHYL-3-OXO-2H-1,4-BENZOXAZINE-2CARBOXYLATE (26)

3.00 g of compound 10 are dissolved in 150 ml of absolute ethanol. The solution was purged with argon and Pd / C was added portionwise (3x100 mg) while blowing. Hydrogen was introduced into the solution (in the Parr reactor) at a pressure of 2 bar and a temperature of 30 ° C. Hydrogenation was carried out for 3 h, since it was shown that the reaction did not proceed quantitatively in half a time. Pd / C is separated by suction filtration. The impure product was recrystallized from ethanol / water (20 ml / 20 ml) and dried on a diaphragm pump.

Yield: 2.30 g (86.0%), melting point: 139.0-141.0 ° C

-4040

IR (KBr, ^cm-1): 3401, 3228, 2986, 1746, 1698, 1658.1518, 1426, 1386, 1316, 1242, 1126, 1009, 843, 616, 573, 470 cm ^-1.

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ (ppm) = 1.08 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.62 (s, 3H, CH ₃ ). 4.07 (q, J = 7.16 Hz, 2H, CH ₂ ), 4.92 (s, 2H, NH ₂ ), 6.17-6.58 (ΑΒΧ, 3H, 3H _ar ), 10.43 (s, 1H, NH).

Example 27:

ETHYL 7 - (((4-CYANOPHENYL) CARBOXAMLDO) -3,4-DIHYDRO-2-METHYL-3-OKSO2H-1,4-BENZOXAZINE-2-CARBOXYLLATE (27)

1,000 g (4.00 mmol) of 26 and 0.56 ml (4.00 mmol) of triethylamine are mixed with 40.00 ml of pre-distilled chloroform and then 0.664 g (4.00 mmol) of 4-ciaobenzoyl chloride are added in two portions. The mixture was allowed to stir at room temperature for 4.5 h, then washed with 2x40 ml of 1 M HCl, 2x40 ml of water and 2x40 ml of saturated NaCl solution. The organic phase was dried with MgSO ₄ and the drying agent was filtered off and the chloroform was evaporated. A precipitate was obtained which did not dissolve in the aqueous phase or in chloroform, and it was dissolved in ethyl acetate and washed with water. Use 150 ml of ethyl acetate and 3x50 ml of water. After separation of the phases, the organic phase was dried overnight with MgSO ₄ , the drying agent was filtered off and the ethyl acetate was evaporated. The residue is taken up in a mixture of ether and hexane and the precipitate is filtered off.

Yield: 1.266 g (83.5%), melting point: 212.0-214.5 ° C

IR (KBr, cm ' ¹ ): 3391, 2231, 1692, 1557, 1518, 1413, 1325, 1131, 1014, 854, 753, 611 ¹ H-NMR (300 MHz, DMSO-d ₆ ): δ (ppm) = 1.08 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.70 (s, 3H, CH ₃ ), 4.07-4.14 (q, J = 7.16 Hz, 2H , CH ₂ ), 6.87-7.56 (ΑΒΧ, 3H, 3H _ar ), 8,008.10 (AB, 4H, 4H ₄ ), 10, 44 (s, 1H, NH), 10.92 (s. 1H, NH)

-4141

M (calculated) = 379,372, found (FAB): 380 (MH ⁺ )

Elemental analysis for C20H17N3O5: calculated found % C 63.32 63.44 % H 4,52 4.66 % N 11.08 11,24

Example 28:

ETHYL 7 - ((4- (AMINO (IMINO) METHYL) PHENYL) CARBOXAMIDO) -3,4-Dihydro-2-METHYL-3-OXO-2H-1.4-BENZOXAZINE-2-CARBOXYLATE (28)

The starting compound 27 (0.500 g (1.21 mmol)) was dissolved in 20 ml of absolute ethanol, cooled on ice and gas HCl was introduced into the solution for 0.5 hours. The reaction mixture was allowed to stir for a further 6 hours. The solvent was evaporated after 6 hours and washed with 3x15 m diethyl ether. The precipitated product was dissolved in 30 ml of absolute ethanol, ammonium acetate (0.256 g (3.32 mmoi)) was added and stirred at room temperature for 24 hours.

Utilization ;; 0.477 g (77.8%), melting point; 193.0-196.0 ° C

IR (KBr, cm ¹ ): 3454.1637, 1560, 1522, 1406, 1118, 863, 590 ¹ H-NMR (300MHz, DMSO-de): δ = 1.08 (t, J = 7.16 Hz. 3H, CH ₃ ), 1.70 (s, 3H, CH ₃ ), 4.08-4.14 (q, J = 7.16 Hz, 2H, CH ₂ ), 6.87-7.58 (ΑΒΧ , 3H, 3H _ar ), 8.09-8.12 (ΑΒ, 4H, 4H ₄ ), 10.43 (s, 1H, NH) ppm.

M (calculated) = 456, found (FAB): 397 (MH ⁺ -acetate)

Elemental analysis for C22H24N4O7.2H2O: calculated found % C 53.70 53.72 % H 5.73 5.63 % N 11,38 11,75

-4242

Reaction scheme for the preparation of compounds 32 and 33:

A: EtBr, benzyltriethylammonium chloride, MeCN, B: Fh, Pd / C, MeOH; C: 4cyanobenzoyl chloride, CHCl _3j D: NH ₂ OH, EtOH; E: 1. HCl (g), EtOH; 2. Ammonium acetate, EtOH

Example 29:

ETHYL N-ETHYL-3,4-DIHYDRO-2-METHYL-7-NTHRO-3-OXO-2H-1,4BENZOXAZINE-2-CARBOXYLATE (29)

Suspend compound 10, 2.466 g (17.85 mmol) of potassium carbonate and 0.813 g (3.57 mmol) of benzyltriethylammonium chloride in 70 ml of acetonitrile. The suspension was stirred and 0.993 g (8.56 mmol) of ethyl bromide was added while stirring. Heat for 60 days at 60 ° C. The precipitate was removed from the reaction mixture and the acetonitrile evaporated. The solid acetonitrile residue was dissolved in toluene.

-4343

The organic phase is washed with 2x25 ml of 0.1 M HCl, then with 2x25 ml of saturated NaHCO ₃ solution and finally with 2x25 ml of saturated NaCl solution. The organic phase was separated, dried with magnesium sulfate and the toluene was evaporated after drying the drying agent.

Yield: 2.01 g (91%), melting point: 75.0-79.0 ° C

IR (KBr, cm ' ¹ ): 2983, 1747, 1715, 1602, 1522, 1391, 1338, 1263, 1140, 1022, 884, 793, 745, 675, 552 ¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1.16 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.32 (t, J = 7.16 Hz, 3H, CH ₃ ) 1.92 (s, 3H, CH ₃ ) , 4.15 (m, J = 7.16 Hz, 1H, CH), 4.17 (q, J = 7.16 Hz, 2H, CH ₂ ), 4.18 (m, J = 7.16 Hz) , 1H, CH), 7.06 (d, 1H, 1H _ar ), 7.98 (2H, 2H _ar ) PPm.

M (calculated) = 308.29, found (ΕΙ): 308

Elemental analysis for C ^ H-jg calculated found

% c 54.54 54.12 % H 5,23 5,28 % N 9.09 9.04

Example 30:

ETHYL N-ETYL-7-AMINO-3,4-DIHYDRO-2-METHYL-3-OXO-2H-1,4BENZOXAZINE-2-CARBOXYLATE (30)

Dissolve 1,800 g (5.84 mmol) of Compound 29 in 100 ml of absolute ethanol. The solution was purged with argon and Pd / C added after blowing

-4444 servings (3x100 mg). H _{2 is} introduced into the solution (with a balloon) at a pressure of 1 bar and at room temperature. Hydrogenation is performed overnight. Pd / C was separated from the solution on the beaker, and the ethanol was evaporated.

Yield: 1,570 g (96.6%), melting point: 105.0-112.0 ° C

IR (KBr, cnT ¹ ): 3467, 3371, 2988, 1727, 1671, 1630, 1513, 1416, 1266, 1124, 1007, 834, 692, 614 ¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1 , 18 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.26 (s, 3H, CH ₃ ), 1.32 (t, J = 7.16 Hz, 3H, CH ₃ ), 3 , 62 (s, 2H, NH ₂ ), 3.87 (m, J = 7.16 Hz, 1H, CH), 4.02 (m, J = 7.16 Hz, 1H, CH), 4.14 (q, J = 7.16 Hz, 2H, CH ₂ ), 6.35-6.78 (ΑΒΧ, 3H, 3H _ar ) ppm.

M (calculated) = 278,308, found (ΕΙ): 278

Elemental analysis for C14H13 N2O4: calculated found % C 60.42 60.59 % H 6.52 6.38 % N 10.07 10,00.

Example 31:

ETHYL 7 - (((4-CYANOPHENYL) CARBOXAMIDO) -N-ETHYL-3,4-DIHYDRO-2-METHYL-3OXO-2H-1.4-BENZOXAZINE-2-CARBOXYLATE (31)

1,200 g (4.31 mmol) of compound 30 and 0.60 ml (4.31 mmol) of triethylamine are mixed in chloroform (40.00 ml) and then 0.716 g (4.31 mmol) is added in two portions.

4-ciaobenzoyl chloride. The mixture was allowed to stir at room temperature for one day and then washed with 2x50 and 1 M HCl, 2x50 ml water and 2x50 ml saturated NaCl solution. The organic phase is dried with MgSO ₄ and the drying agent is filtered off,

-4545 chloroform was evaporated. The compound was left overnight in cold methanol and the precipitated crystals were filtered off by suction.

Yield: 1,004 g (57.1%), melting point: 143.0-147.0 ° C

IR (KBr, cm ' ¹ ): 3293, 2982, 2228, 1756, 1696, 1655, 1514, 1395, 1326, 1188, 1130,1017,863,763 ¹ H-NMR (300 MHz, CDCI ₃ ): δ = 1. 17 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.29 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.86 (s, 3H, CH ₃ ), 3. 92 (m, J = 7.16 Hz, 1H, CH), 4.07 (m, J = 7.16 Hz, 1H, CH), 4.14-4.17 (q, J = 7.16 Hz , 2H, CH ₂ ), 7.00-7.46 (ΑΒΧ, 3H, 3H _ar ), 7.82-7.97 (AB, 4H, 4H ₄ ) ppm.

M (calculated) = 407,426, found (FAB): 408 (MH ⁺ )

Elemental analysis for C22H21N3O5: calculated found % C 64.86 64.96 % H 5.20 5.46 % N 10,31 10,01

Example 32:

ETHYL 7 - ((4- (AMINO (HYDOXYIMINO) METHYL) PHENYL) CARBOXAMIDO) -N-ETHYL 4,4-DIHYDRO-2-METHYL-3-OXO-2H-1,4-BENZOXAZINE-2-CARBOXYLATE (32)

Dissolve 0.205 g (2.95 mmol) of hydroxylammonium chloride in absolute ethanol (25.00 ml), add 0.299 g (2.95 mmol) of triethylamine and stir for 20 minutes at room temperature. To the solution was added 0.300 g (0.74 mmol) of compound 31 and stirred at 50 ° C for 96 hours. The ethanol was evaporated and the solid residue was taken up with acetone. The precipitate in acetone is filtered off by suction.

Yield: 0.251 g (77.0%), melting point: 105.0-109.0 ° C

-4646

IR (KBr, ^cm-1): 3349, 2979, 1750, 1670, 1514, 1403, 1258, 1124, 1015, 858, 810, 803 ¹ H-NMR (300 MHz, CDCI _3): δ = 1.15 ( t, J = 7.16 Hz, 3H, CH ₃ ), 1.26 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.83 (s, 3H, CH ₃ ), 3.85 ( m, J = 7.16 Hz, 1H, CH), 4.05 (m, J = 7.16 Hz, 1H, CH), 4.09-4.11 (q, J = 7.16 Hz, 2H , CH ₂ ), 5.36 (s, 2H, NH _Z ), 6.92-7.44 (ΑΒΧ, 3H, 3H _ar ), 7.65-7.77 (AB, 4H, 4H ₄ ), 8 , 36 (s, 1H, NH) ppm.

M (calculated) = 440,456, found (FAB): 441 (MH ⁺ )

Elemental analysis for C22H24 N4O6 H2O: calculated found % C 57.63 57.53 % H 5.72 6.03 % N 12,22 11,78

Example 33:

ETHYL 7 - ((4- (AMINO (IMINO) METHYL) PHENYL) CARBOXAMLDO) -N-ETHYL-3,4-Dihydro-2-METHYL-3-OXO-2H-1.4-BENZOXAZINE-2-CARBOXYLATE (33)

The starting compound 32 was dissolved in absolute ethanol (30 ml), cooled on ice and gas HCl was introduced into the solution for 0.5 hours. The reaction mixture was allowed to stir for a further 24 hours. The solvent was evaporated after 24 hours and washed with 3x15 ml diethyl ether. The precipitated product was dissolved in 30 ml of absolute ethanol, 0.171 g (2.22 mmol) of ammonium acetate was added and stirred at room temperature for 24 hours. The ethanol was evaporated and the solid residue was poured over with acetone / ethanol = 3/1. A white precipitate appears, which we drain. The product is a pale brown amorphous, very hygroscopic substance.

Yield: 0.151 g (48.1%), melting point: 146.0-158.0 ° C

-4747

IR (KBr, ^cm-1): 3136, 3046, 1737, 1679, 1606, 1548, 1514, 1407, 1317, 1280, 1123, 1011,861,705, 649 ^cm-first

¹ H-NMR (300 MHz, DMSO-d ₆ ): δ = 1.05 (t, J = 7.16 Hz, 3H, CH ₃ ), 1.15 (t, J = 7.16 Hz, 3H. CH ₃ ), 1.73 (s, 3H, CH ₃ ), 3.37-3.42 (broad signal, probably due to the hygroscopicity of the compound overlapping siginamidine), 3.85 (m, J = 7.16 Hz, 1H, CH), 4.06-4.08 (q, J = 7.16 Hz, 2H, CH ₂ ), 4.36 (m, J = 7.16 Hz, 1H, CH), 7.227. 67 (ΑΒΧ, 3H, 3H _ar ), 7.94-8.13 (AB, 4H, 4H ₄ ), 10.54 (s, 1H, NH) ppm.

M (calculated) = 484,509, found (FAB): 425 (MH ⁺ - acetate)

Example 34:

RESULTS OF COMPOUND ACTIVITY TESTING

Table 1. IC50 for certain peptidomimetic GPIIb / IIIa antagonists subject to patent application. The antagonists are classified according to the inhibition of ADP-induced aggregation compared to the natural figand for this RGDS receptor (Arg-Gly-Asp-Lys).

[compound label and i and i and i and i I ΐ i ADP (20 μΜ) collagen (10 μΜ) adrenaline (10 μΜ) Aggregation (IC50 (μΜ)) speed aggregation (IC50 (μΜ)) aggregation I (IC50 (μΜ)) speed aggregation (IC50 (μΜ)) aggregation (IC50 (μΜ)) RGDS 260 ± 52 i 70 ± 5 410 ± 30 150 ± 30 210 ± 30 16 0.9 9.2 1.1 - 5 3 3 ......................... 42 Γ .................. 20 - 25 5 .......................... 21 35 ............................... 16 ..... - 9 : 30 s 15 35 15 - ..... 33 .................. ....... ~ 44 .................... 42 ..... 189 63 - ; 2S ! 45 24 Ϊ85 and 48 I 10

Claims (6)

PATENT APPLICATIONS 1. Compounds of general formula (I): 0) where it means: A is an O, S, N or methylene group, B is C or N, E and D are H or a chain of the following structure: Xa- (CH ₂ ) _n -Y-, where where R represents H, OH, COORi or COR1, R4 O or S and R ₅ H, alkyl branched or unbranched C1-C7 chain, -4949 a is a CO or 5-7 membered alicyclic or aromatic ring system which, in addition to O, may contain 1 or 2 O or N heteroatoms, n is 0-4, Y is O, -NH-CO-, -CO-NH-, NH-CO-NH-, -NH-, -NH-CH ₂ -, -OH ₂ -NH-, -CH ₂ -NH-CO-, - CO-NH-CH ₂ -; wherein one of E and D is always H and the other is a chain; Ri is H or alkyl branched or straight chain Ci-C _4, R ₂ isH, COOR _{1 (} -CONHR ₆ , -NHCOR7, -CH ₂ -COORi, -CH ₂ -CONHR ₆ , -O-CH ₂ -COOR _1f -O-CH ₂ -CO NHR _6j CO where R _{6 is a} group of the formula wherein R 8 is H, NH-SO ₂ R 10, a branched or unbranched alkyl chain of C 1 -C 4 atoms, a benzyl group, Rg is H or a branched or unbranched C1-C4 alkyl chain, R10 is an aliphatic group Οι-Οθ or wherein Rn, -CH3, -NH ₂ or -NHCOCH3, R7 is a benzene ring which is substituted at the p- or m-position with a group X, -5050 R ₃ is H, a branched or unbranched alkyl group of C 1 -C 4 atoms, - (CH 2) n, -COOR 1 or - (CH ₂ ) n-X their enantiomers, diastereoisomers, mixtures of diastereomers, racemates and pharmaceutically acceptable salts. Compounds of formula I according to claim 1 for use as therapeutically effective substances. Use of compounds of formula I according to claim I for the preparation of medicaments that inhibit platelet aggregation induced by the binding of fibrinogen to the fibrinogen receptor on platelets. Use of the compounds of formula I according to claim 1 for the preparation of medicaments for use in the prevention and treatment of arterial thrombosis, ischemic stroke, peripheral arterial disease, acute coronary syndrome and for the prevention of ischemic events in patients with surgery. Pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula I according to claim 1 and pharmaceutically acceptable excipients. Pharmaceutical preparations according to claim 5, characterized in that they are used to inhibit platelet aggregation in human blood and other mammals through antagonistic action on the fibrinogen receptor.