SI21137A - Derivatives of azaphenyl-alanine - Google Patents

Abstract

New derivatives of azaphenyl-alanine having formula I and their pharmaceutically acceptable salts are described, where their substituents have meanings given in the description. The compounds are used as anticoagulants.

Description

FIELD OF THE INVENTION

The invention is within the scope of the pharmaceutical industry and relates to novel azaphenylalanine derivatives, their use, processes for their preparation and pharmaceutical products containing them.

New azaphenylalanine derivatives are used as anticoagulants.

A technical problem

There is a continuing need for new anticoagulant agents, especially those that would be orally active, as selective as possible, and with as few side effects as possible.

The state of the art

Thrombin, as a serine protease, is one of the key enzymes in the processes of blood clotting and the development of thrombosis (Edit, JF; Allison, P.; Noble, S.; Ashton, J.; Golino, P.; McNard, J.; Buja, LM; Willerson, JT, J. Ciin. Invest. 1989, 84, 18.).

The crystal structure of the serine proteases of thrombin and serine is known (Bode, W.; Turk, D.; Karshikov, A. J., Protein Sci. 1992, 1, 426.

The anticoagulants used now have many side effects and limited efficacy.

Low molecular weight thrombin inhibitors are expected to have selective efficacy and the potential for oral administration. Reversible and irreversible thrombin antagonists are known (Kimball, SD, Curr. Pharm. Design 1995, 1, 441; Das, J.; Kimball, SD, Bioorg. Med. Chem. 1995, 3, 999, Kimball, SD, Blood Coagulation and Fibrinolysis 1995, 6, 511, Breznik, M.; Pečar, S., Farm, West 1997, 48, 545; Leung, D.; Abbenante,

G.; Fairlie, D. P., J. Med. Chem. 2000, 43, 305).

Most reversible thrombin antagonists are derived from the peptidomimimetically modified structure of D-Phe-Pro-Arg. The aim of the changes is to ensure chemical stability, selectivity and efficiency.

As a medicament, the active ingredient argatroban is described in EP 8746. Napsagatran is a selective reversible inhibitor described in EP 559046. Meta substituted phenylalanine derivatives with an amidine or amidoxime structure are selective thrombin inhibitors known from WO 92/08709. The amidinophenylalanine derivatives and the aminopyridylalanine derivatives described in WO 95/13274 have selective antithrombotic activity.

Azapeptides are peptidomimetics in which the Ca atom is replaced by nitrogen. The advantage of such isoelectronic substitution is the preservation of the chemical integrity of the altered amino acid and only a small conformational change, which is important for the process of molecular recognition and ligand-receptor complex stabilization and metabolic stabilization (Gante, J., Synthesis 1989, 405; Gante, J., Angew. Chem. 1994, 106, 1780).

The aza-phenylalanine fragment thrombin inhibitor group described in SI-20025 had submichromolar inhibitory constants in in vitro assays.

The preparation of N-naphthylsulfonyl amino acids (Pendleton.RG, et al. J. Pharm. Exp. Ther. 1979, 208, 24) and 1,2,3,4-tetrahydroisoquinoline derivatives are known in the preparation of novel thrombin inhibitors (Pendelton , RG; Gessner, G.; Weiner, G.; et al., J. Pharm. Exp. Ther. 1978, 208, 24).

Description of solution to a technical problem with implementation examples

The invention relates to novel azaphenylalanine derivatives and their analogs of the general formula (I)

R ¹

in which they mean

NH ₃ ⁺ Cl '

R ^{1 is a} radical of formula wherein R ⁴ is H, alkyl (C 1 -C ₃ ), OH, O-alkyl (C 1 -C ₃ ), NH ₂ ;

R ^{2 is a} residue of the formula

where R ⁵ = H, alkyl (C 1 -C 3), COOR ¹⁰ ,

R ⁶ = H, alkyl (0τ-O ₃ ), COOR ¹⁰ ,

R ⁷ = H, alkyl (C 1 -C 3), COOR ¹⁰ ,

R ¹⁰ = H, alkyl (C _r C ₃ ),

R ⁸ = H, alkyl (C 1 -C 3), cycloalkyl (C 3 -C 6), R ⁹ = H, alkyl (C 1 -C 3), cycloalkyl (C 3 -C ₆ ), R ¹¹ = H, alkyl (C 1 -C 3) , benzyl,

X = CH, O, S,

Y = NR ¹² , O, S,

R ¹² = H, COCH ₃ , alkyl (C 1 -C 6);

R ^{3 is a} residue of the formula

/ ^NH 3 ^C 'in the case where R - Γ

The invention also relates to their pharmaceutically acceptable salts with anticoagulant activity and to the pharmaceutical preparations containing them. The invention also relates to methods for the preparation of azaphenylalanine derivatives and analogs of general formula (I).

Azaphenylalanine derivatives of general formula (I), when R ¹ = ^H2N ^ - ^NR "> are prepared by

a) 4-cyanobenzaldehyde of formula (II)

CN (H)

CHO is converted with the BOC-carbazate of formula (III)

(Hi) to a compound of formula (IV)

CN

Yy

I n

N '

I

NH (IV) which is converted into a compound (V) (V) by a catalytic hydrogenation reduction process

HN I

CL .NH reacting with triphosgene and an amine of formula (VI)

R ⁵ R ⁶

HN

R ⁷ (VI) wherein R ⁵ , R ⁶ and R ^{7 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (VII or Vlyl),

(Vil) (Vlil) or with triphosgene and an amine of formula (IX)

Γ

HN.

^{R 9} (IX) wherein R ⁸ and R ^{9 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (X),

where R ^{11 has the} same meaning as in formula (I), to compound (XI)

where R ^{2 has the} same meaning as in formula (I).

The BOC protecting group in compound (XI) is removed by HCl (g) in HOAc at room temperature to give compound (XII),

CN (XII)

NH * ci '

J where R ^{2 has the} same meaning as in formula (I).

Compound XII is then reacted with an aromatic sulfonyl chloride or naphthylsulfonyl amino acid with an activated carboxylic group to compound (XIII)

(XIII)

The compound (XIII) of formula (XIV) and R ^{3 have the} same meaning as in formula (I).

is reacted with hydroxylamine in anhydrous ethanol to compound with (XIV)

R ¹

where R ¹ = OH.

, R ² and R ^{3 have the} same meaning as in formula (I), and R ⁴ represents

The compound (XIII) is reacted by introducing gaseous HCl in methanol solution, adding ammonium acetate and reintroducing HCl into compound (XV) where R ¹ = ^H 2 ^{N 1} - ^{NR 4} , r ² j _n r ³ have the same meaning as in formula ( I), and R ⁴ represents hydrogen.

R ¹

Azaphenylalanine derivatives of general formula (I), when R ¹ = | / ^NH 3 ^CI , are prepared by

a) 4-cyanobenzaldehyde of formula (II)

CN

CHO is converted with ethylene glycol in the presence of 4-toluenesulfonic acid to compound (XVI)

CN

O O _ which is reduced by lithium aluminum hydride to a compound of formula (XVII)

o o reacting with acetanhydride to compound (XVIII)

The compound (XVIII) with 90% methanoic acid is converted into compound (XIX) ch ₂ nhcoch ₃

CHO (XIX) converted by BOC-carbazate of formula (III) / '' (lil)

I o to a compound of formula (XX)

CH ₂ NHCOCH ₃ (XX)

which is converted by the catalytic hydrogenation reduction process to compound (XXI) ch ₂ nhcoch ₃

J (XXI)

HN γν ^{'Η 1} pp

which reacts with triphosgene and an amine of formula (VI)

wherein R ⁵ , R ⁶ and R ^{7 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (VII or VIII),

(Vil) (Vlil) or with triphosgene and an amine of formula (IX)

Γ

HN.

^RS (IX) wherein R ⁸ and R ^{9 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (X),

wherein R ^{11 has the} same meaning as in formula (I) to compound (XXII)

-1010 where R ^{2 has the} same meaning as in formula (I).

The BOC protecting group in compound (XXII) was removed by HCl (g) in AcOH at room temperature to give compound (XXIII), ch ₂ nhcoch ₃ (XXIII)

I.

NH ₃ Cl wherein R ^{2 has the} same meaning as in formula (I) which reacts with aromatic sulfonyl chloride to compound (XXIV) ch ₂ nhcoch ₃ o

wherein R ² and R ^{3 have the} same meaning as in formula (I).

Compound (XXIV) is digested by boiling in 5M HCl to compound (XXV)

and R ³ have the same meaning as in formula (I).

The starting compounds are prepared, unless otherwise stated, according to the procedures described in the literature; e.g. a compound of formula IV as described by A. Fassler, et. al., J. Med. Chem. 1996, 39, 3203-3215.

-1111

The invention also relates to the use of compounds of formula I as oral and parenteral anticoagulants. They can be used in the treatment and prevention of various forms of thrombosis: (i) venous thromboembolism in which thrombus is formed in a vein (vein thrombosis) due to acquired (lying, surgery, injury, malignancy, pregnancy and childbirth) or congenital (defects in natural coagulation inhibitors) risk factors, and the torn part of the thrombus narrows or closes the pulmonary arteries (pulmonary embolism), (ii) in cardiogenic thromboembolisms that cause thrombus in the heart due to disorders of the heart rhythm, failure of the heart valves, artificial heart valves, or heart muscle disease, and causes emboli in the peripheral arteries, most commonly in the brain (ischemic stroke), (iii) arterial thrombosis, which occurs on the atherosclerotic altered artery and narrows or closes the artery, and causes cardiac muscle ischemia (angina, acute coronary syndrome) or death heart attack), narrow or close the peripheral artery (ischemic disease of the peripheral arteries ) and narrow or close the artery after the intervention on the vein (reocclusion or restenosis after percutaneous transluminal coronary angioplasty, reocclusion or restenosis after percutaneous transluminal angioplasty of the peripheral arteries) and (iv) in conditions (n. BC complications in pregnancy, in metastatic malignancies, after extensive injuries, in bacterial sepsis), when a thrombogenic stimulus causes diffuse thrombus formation in the vascular system (disseminated intravascular coagulation).

The compounds of formula I may also be used as adjunctive therapy in combination with thrombolytic therapy for fresh heart attack, in combination with aspirin in patients with unstable angina who will undergo percutaneous transluminal angioplasty and in the treatment of patients with thrombosis and heparin-induced thrombocytopenia.

Anticoagulation medicines are also used to prevent blood clotting that comes in contact with non-biological surfaces (vascular prostheses, blood vessels, artificial heart valves, extracorporeal blood systems, hemodialysis) and in vitro when preparing biological samples for testing or storage.

-1212

Also contemplated are pharmaceutical preparations containing compounds of formula I. They may be in the form of various injections or oral preparations. In addition to the active substance, they contain various standard additives depending on the type of use. Pharmaceutical preparations may be prepared by standard procedures. They can also be formulated in various forms to ensure controlled and prolonged release of the active substance. The dose, frequency and method of administration depend on various factors and depend on the individual substance and its pharmacokinetic properties and the patient's condition.

BIOLOGICAL TESTS

I. Enzyme Method

1. Determination of efficacy of thrombin inhibitors

a) Principle

Thrombin cleaves amide bonds in a synthetic chromogenic substrate, releasing yellow-colored p-nitroaniline (p-NA). The amount of released p-NA is proportional to the absorbance measured with a spectrophotometer at a wavelength of 405 nm. With the addition of a thrombin inhibitor, the amidolytic activity of the enzyme is reduced. The inhibitor efficiency is expressed by the inhibition constant (K,).

b) Reagents

Thrombin (human thrombin, 308 NIH units, Sigma): Dissolve the contents of the bottle with distilled water to obtain a stock solution of 20 NIH units / mL. This was pipetted over 0.5 mL and stored at -70 ° C. HBSA buffer was prepared immediately before use with a working solution of thrombin with an activity of 2 NIH units / mL. The final concentration of thrombin in the microtiter plate is 0.5 NIH units / mL.

Chromogenic substrate for thrombin (S-2238, Chromogenix, 25 mg). Prepare a 1 mM substrate solution, pipette 0.5 mL and store at -20 ° C. Before use

-1313 is prepared with distilled water of 160 and 80 μΜ of substrate solution. The final concentrations of the substrate in the reaction mixture are 40 oz. 20 μΜ (Km = 2.6 μΜ).

HBSA buffer, pH 7.5: 10 mM Hepes buffer (HEPES, Sigma), 150 mM NaCl and 0.1% (w / v) bovine serum albumin (98% bovine serum albumin, Sigma) were dissolved in bidestilled water. The pH was adjusted with a 0.1 M NaOH solution.

Inhibitors: Dissolve them with DMSO to give 10 mM stock solution. Working solutions (final concentrations of 10 to 100 μΜ) are prepared with distilled water. The maximum concentration of DMSO in the microtiter plate did not exceed 3%.

c) Workflow

The measurements are carried out in a microtiter plate. Pipette 50 μΙ_ HBSA buffer, 50 μΙ_ inhibitor solution of different concentrations (in case of control 50 μΙ_ HBSA buffer) and 50 μΙ_ thrombin solution into the wells of the microtiter plate. The mixture in the plate was incubated for 15 minutes at 25 ° C. After incubation, 50 μm of the chromogenic substrate was added and the microtiter plate was inserted into a spectrophotometer (Tecan, Sunrise). We measure the increase in absorbance at a wavelength of 405 nM and a temperature of 25 ° C for 15 minutes every 10 seconds.

40 and 20 μΜ substrate are used to determine the inhibition constant (Kj). Each measurement is made in triplicate and the average value of the three measurements is taken into account for the calculation of the result.

d) Determination of the inhibition constant (K,)

Kj is determined according to the principle described by Cheng and Prusoff (Biochem Pharmacol, 1973). The initial reaction rates in the presence and in the absence of an inhibitor are measured. The change in absorbance in time unit (v) is calculated from the initial, linear part of the reaction. Competitive inhibitors are said to be ^v ; _ K, + S and it follows that

-1414 ^K '(h /^.)+ O' ((v '/ v,) - l)

I = inhibitor concentration, S = substrate concentration, Km = Michaelis constant, v ₀ = initial reaction rate without inhibitor, v, = initial reaction rate in the presence of inhibitor.

Measurements are made with two concentrations of inhibitor and two concentrations of substrate. For each combination of substrate and inhibitor concentrations used, K is calculated and their transverse value is taken as a result.

2. Determination of selectivity of thrombin inhibitory activity with respect to trypsin inhibition

a) Principle

Due to the similarity of thrombin and trypsin to substrate specificity resulting from the comparable structure of the active site, we determine the selectivity of the inhibitory activity against thrombin relative to the inhibition of trypsin, which is a non-specific serine proteinase. The inhibitory activity against thrombin is determined as described above. Trypsin inhibition is measured in the same way as thrombin inhibition, but another chromogenic substrate is used. For both enzymes we calculate K. From the K ratio for trypsin / K, for thrombin we deduce the inhibitor selectivity.

b) Reagents

Trypsin (bovine, 6000 BAEE units / mg protein, Sigma): Prepare a trypsin stock solution with an activity of 300 Ε / mL, pipette 0.2 mL and store at -70 ° C. Immediately before use, thaw the stock solution and prepare with HBSA buffer working solution with activity of 4 mE / mL. The final trypsin activity in the microtiter plate is 1 mE / mL.

Chromogenic trypsin substrate (S-2222, Chromogenix, 25 mg): Prepare a 2 mM substrate solution, pipette 0.3 mL each and store at -20 ° C. Before use

The -1515 stock solution is thawed and 400 and 200 μΜ of substrate solution are prepared. The final concentrations of the substrate in the reaction mixture are 100 oz. 50 μΜ. (Km = 25 µΜ)

HBSA buffer, pH 7.5: same as the thrombin method

Inhibitors: Dissolve them with DMSO to give 10 mM stock solution. Working solutions (final concentrations of 10 to 600 μΜ) are prepared with distilled water. The maximum concentration of DMSO in the microtiter plate did not exceed 10%.

100 and 50 μΜ of substrate are used to determine K. We perform each measurement in triplicate and take the average value of these three measurements as a result,

c) Workflow

The workflow is the same as that described when measuring thrombin inhibitory activity. The concentrations of reagents described to determine the trypsin inhibitory activity are used.

d) Determination of the inhibition constant (K)

Proceed as described in determining K for thrombin.

e) Determining selectivity

We determine K for thrombin and K for trypsin. We define selectivity as a ratio:

K .- (trypsin) selectivity = —--—

K. (thrombin)

3. Determination of selectivity of thrombin inhibitory activity with respect to factor Xa inhibition

a) Principle

The selectivity of inhibitory activity against thrombin relative to the inhibition of factor Xa is determined by the relatedness of both enzymes and the similar structure of the active site. The inhibitory activity against thrombin is determined as described above. FXa inhibition is measured in the same way as inhibition of thrombin by a chromogenic substrate

-1616

S2238. For both enzymes we calculate K,. From the Kj ratio for FXa / K, for thrombin we deduce the inhibitor selectivity.

b) Reagents

Factor Xa (Chromogenix, 71 nkat): Dissolve the contents of the bottle with distilled water to give a stock solution of 10 nkat / mL. Pipette this at 0.5 ml_ and store at -20 ° C. With HBSA buffer, a working solution of FXa with an activity of 2 nk / mL was prepared immediately before use. The final activity of FXa in the microtiter plate is 0.5 nkat / mL.

Chromogenic substrate for FXa (S-2222, Chromogenix, 25 mg). Prepare a 2 mM substrate solution, pipette 0.5 mL and store at -20 ° C. Prepare 800 and 400 μΜ of substrate solution with distilled water before use. The final concentrations of the substrate in the reaction mixture are 200 oz. 100 μΜ (K _m = 164 μΜ).

HBSA buffer, pH 7.5: same as the thrombin method

Inhibitors: Dissolve them with DMSO to give 10 mM stock solution. Working solutions (final concentrations of 5 to 300 μΜ) are prepared with distilled water. The maximum concentration of DMSO in the microtiter plate did not exceed 3%.

c) Workflow

The workflow is the same as that described when measuring thrombin inhibitory activity. The concentrations of the reagents described to determine the inhibitory activity relative to FXa are used.

d) Determination of the inhibition constant (K)

Proceed as described for the determination of K for thrombin.

e) Determining selectivity

We determine K for thrombin and K for FX. We define selectivity as a ratio:

selectivity =

Kj (FXa) Kj (thrombin)

-1717

II. Coagulation investigations

Coagulation tests (thrombin time, activated partial thromboplastin time, and prothrombin time) measure the solidification time of normal mixed plasma in a coagulation apparatus after adding different concentrations of inhibitor. The results are expressed as the concentration of inhibitor, which prolongs the curing time by two times.

1. Thrombin time (PT)

a) Principle

The determination of thrombin time is used in the laboratory control of unfractionated heparin treatment, in the control of thrombolytic therapy, in the detection of fibrin production disorders and in the detection of severe fibrinogen deficiency. Thrombin time is prolonged when fibrinogen concentration is decreased, when fibrinogen degradation products or thrombin inhibitors are present. The procedure is such that thrombin is added to the plasma. Thrombin converts fibrinogen into fibrin, which is identified as the formation of a clot.

b) Reagents

Thrombin (Thrombin Reagent Test, 1.5 IU / mL): Dissolve lyophilized bovine thrombin in 5 mL of HEPES buffer (25 mM, pH 7.4).

Normal mixed plasma '. At least 10 apparently healthy subjects are donated venous blood to a 0.11 M sodium citrate solution (1 part sodium citrate and 9 parts blood). Centrifuge the blood immediately after collection at 2000 x g for 30 minutes and 4 ° C. Plasma was removed, 2 ml was pipetted and stored at -70 ° C.

Inhibitors: Dissolve the inhibitors in DMSO (10 mM stock solution) and dilute with distilled water to obtain working solutions (maximum concentration 100 μΜ)

-1818

c) Workflow

90 μΙ_ of plasma and 10 μL of inhibitor were pipetted into a coagulation apparatus cell (Fibrintimer, Dade / Behring) heated to 37 ° C. Incubate for 5 min. at 37 ° C. Add 200 μΙ_ of thrombin solution heated to 37 ° C. After the addition of thrombin, we turn on the measurement of time, which is measured until the formation of a clot.

2. Activated partial thromboplastin time (APTC)

a) Principle

APTC is used as a screening method to detect interference with the intrinsic part of the coagulation system. The method is sensitive to the shortcomings of the Vlil and IX coagulation factors and contact factors. In the absence of factors or in the presence of inhibitors (eg, lupus anticoagulants, heparin), prolonged APT is measured. It is done by incubating the plasma with the optimal amount of phospholipids and contact activator to activate the factors of the intrinsic coagulation system. The addition of calcium triggers the coagulation process. We measure the time to fibrin formation.

b) Reagents

Phospholipids with activator (Pathromtin SL, Dade / Behring): silica particles, plant phospholipids, sodium chloride (2.4 g / L), HEPES (14.3 g / L), pH

7.6 and sodium azide (<1 g / L). It must be at room temperature and well mixed before use.

Calcium chloride: 0.025 mol / L

Normal mixed plasma: Same as thrombin time.

Inhibitors: As with thrombin time.

c) Workflow

Pipette 90 μί of plasma and 10 pL of inhibitor into a coagulation apparatus cell (Fibrintimer, Dade / Behring) heated to 37 ° C. Incubate for 5 min. at 37 ° C.

-1919

Add 100 μl phospholipids with activator. Incubate for 2 min. at 37 ° C. Add 100 µL of calcium chloride at 37 ° C. After the addition of calcium chloride, we begin to measure the time until the clot forms.

3. Prothrombin time (MS)

a) Principle

Prothrombin time is a rapid and sensitive screening method for detecting disturbances in the extrinsic part of the coagulation system (factors II, V, VII and X). Due to the high sensitivity to these coagulation factors, the method is particularly suitable for monitoring oral anticoagulation treatment, for identifying inherited and acquired disorders of these factors, and for checking the synthetic ability of the liver. Prothrombin time is prolonged in the absence of coagulation factors and in the presence of inhibitors. This is done by adding the optimum amount of thromboplastin and calcium to the plasma and measuring the time to clot formation.

b) Reagents

Thromboplastin (Thromborel S, Dade / Behring): Calcium chloride-based human thromboplastin with stabilizers; dissolved in 4 mL of distilled water. Before use, heat the reagent for at least 30 minutes at 37 ° C.

Normal mixed plasma: Same as thrombin time.

Inhibitors: As with thrombin time.

c) Workflow

90 pL of plasma and 10 pL of inhibitor were pipetted into a coagulation apparatus cell (Fibrintimer, Dade / Behring) heated to 37 ° C. Incubate for 5 min. at 37 ° C. Add 200 µL of thromboplastin heated to 37 ° C. After the addition of thromboplastin, we begin to measure the time to clot formation.

-2020

Results of bioassays

Compound Ki - thrombin (μΜ) Ki - trypsin (μΜ) Ki - factor X (μΜ) MS (s) Point (s) APTCH (s) 1 6.9 2 12.6 3 10.3 4.9 769 591 396 4 9.0 6.2 37.0 5 8.6 2.5 6 4.0 23.4 59.5 7 23.0 65.5 8 21.0 30.0 63.6 364 1238 893 9 0.92 13 73.9 92 30 893 10 1.3 39.4 76.2 60 25 63 11 1.6 6.9 55.7 202 47 140 12 3.3 13.6 41.1 218 72 158

Compound 1

Compound 2

-2124

Compound 3

Compound 4

Compound 5

-2212-

-23 Compound 8

N

-24Compound 11

NHo Cl '

Compound 12

H ₃ C

ch ₃

Nhk Cl '

-25The invention is explained but by no means limited by the following embodiments.

EXAMPLE 1- {2- [2- (1-Azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazino] -2-oxoethyl} -2-naphthalenesulfonamide

1.00 g (3.77 mmol) of 2 - [(2-naphthylsulfonyl) amino] acetic acid were dissolved in 20 ml of dichloromethane and 420 mg (3.87 mmol) of ethyl chloroformate and 516 mg (4.00 mmol) of N-ethyl-N, N- were added with stirring. diisopropylamine. The reaction mixture was stirred for 1 hour at room temperature, then powdered 2- (1-azepanylcarbonyl) -2- (4cyanobenzyl) hydrazinium chloride (1.00 g; 3.24 mmol) was added. After addition, it was stirred for 3 days at room temperature.

Remove the solvent from the rotavapor and dissolve the residue in 50 ml of ethyl acetate and then wash with 1M HCl, 1M NaOH and distilled water. The organic phase was dried with sodium sulfate and the solvent removed under reduced pressure. The crude product was recrystallized from ethanol.

Efficiency; 1.34 g (79%)

Melting point: 149-151 ° C

IR (KBr, cm ^-1 ); 3410, 3253, 2919, 2232, 1708, 1613, 1440, 1343, 1156, 659 ¹ H-NMR (DMSO-de): δ (ppm) 1.46 (m, 8H, CH 2 -azepine ₃ ', 4', 5. 6 '); 1.65 (m, 4H, CH ₂ azepine ₂ ', ₇ '); 3.33 (m, 2H, CO-CH ₂ -NH); 4.09 (m, 1H, NHCO); 4.47 (s, 2H, Ar-CH ₂ ),

7:58 and 7.79 (2d, 2H each, J _{2, 6} = 8.27, J _3i5 = 27.8, Ar-H, _{2, 3, 5, 6);} 7.69-8.47 (m, 7H, Ar-H (naphthalene)); 10.29 (s, 1H NHSO ₂ )

Molecular Weight: calculated: 519 (C27H29N5O4S); found; 520 (MH ⁺ )

EXAMPLE 2

-264 - ((1- (1-azepane and I carbonyl I) -2- (2 - ((1-naphthyl Isu Ifon and I) ammonium acetate) methyl] - / Hydroxybenzenecarboximidamide / \ / - {2- [2- (1-Azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazino] -2-oxoethyl} -2-naphthalenesulfonamide (400 mg. 0.77 mmol) was dissolved in anhydrous ethanol and hydroxylamine (28.0 mg, 0.85 mmol) was added. The reaction mixture is heated at reflux for 12 hours.

Remove the solvent from the rotary evaporator and wash the product with ether.

Yield: 386 mg (87%)

Melting point: 104-108 ° C

IR (KBr, cm ' ¹ ): 3359, 2624, 1782, 1640, 1422, 1345, 1157, 1075, 750, 661 ¹ H-NMR (CDCl ₃ ): 6 (ppm) 1.49-1.89 (m, 8H, CH _2- azepine ₃ , 4,5,6 '); 2.07 (m, 4H, CH ₂ azepine ₂ , ₇ ); 3.48 (m, 2H, CO-CH ₂ -NH); 4.18 (s, 1H, NHCO); 4.68 (m, 2H, Ar-CH ₂ ), 4.90 (s, 1H, = N-OH); 7.42 and 7.61 (2d, 2H each, J ₂ , ₆ = 8.17, J ₃ , ₅ = 8.15, Ar-H ₂ , ₃ , ₅ , ₆ ); 7,648.34 (m, 7H, Ar-H (naphthalene)); 8.42 (s, 2H, NH ₂ -amidoxime); 10.35 (s, 1H NHSO ₂ ) Molecular weight: calculated 552 (C ₂ 7H ₃₂ N ₆ O ₅ S), measured 553 (MH ⁺ )

EXAMPLE 3

Amino {4 - [(1- (1-azepanylcarbonyl) -2- {2 - [(1-naphthalenesulfonyl) amino] acetyl} hydrazino) methyl] phenyl} methaniminium chloride / N- {2- [2- (1-Azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazino] -2-oxoethyl} -2naphthalenesulfonamide (520mg, 1.00 mmol) was dissolved in 25 ml of anhydrous ethanol and HCI gas was introduced for 20 min. After complete administration, the solution was stirred for 4 hours at room temperature, then ammonium acetate (85.0 mg, 1.10 mmol) was added. The reaction mixture was left at room temperature for 2 days. We reintroduce HCI gas for 20 min. After 1 hour, the resulting ammonium chloride was filtered off and the ethanol was evaporated on a rotary evaporator. The product was poured into ether and filtered off by suction.

Yield: 481 mg (84%)

IR (KBr, cm ' ¹ ): 2969, 2758, 1783, 1669, 1344, 1158, 1076, 749, 660,

-272 Τ ¹ H-NMR (DMSO-d ₆ ): δ (ppm) 1.57-1.76 (m, 8H, CH ₂ -azepine ₃ ', ₄ ', ₅ ', 6'); 3.03 (m, 4H, CH ₂ -azepine ₂ ', 7'); 3.37 (m, 2H, CO-CH ₂ -NH); 4.31 (m, 1H, NHCO); 4.74 (s, 2H, ArCH ₂ ), 7.62 and 7.90 (2d, 2H each, J ₂ , ₆ = 8.14, J ₃ , ₅ = 8.19, Ar-H ₂ , ₃ , ₅ , ₆ ); 7.35-8.49 (m, 7H, Ar-H (naphthalene)); 9.30 (s, 1H NHSO ₂ ); 9.42 (2s, 2H, amidine)

Molecular Weight: Calculated: 537 (C ₂ 7H ₃₃ N ₆ O ₄ S); found: 538 (MH ⁺ )

EXAMPLE 4

2-806 N - N - (1-Azepanylcarbonyl) - N - (4-cyanobenzyl) -1,2,3,4-tetrahydro-7 isoquinolinsulfonohydrazide

2- (1-Azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazinium chloride (2.01 g, 6.51 mmol) was dissolved in 30 ml of dichloromethane. 900 mg (6.98 mmol) of N-ethyl-N, Ndiisopropylamine and 1.93 g (8.00 mmol) of 2-acetyl-1,2,3,4-tetrahydro-7-isoquinolinsulfonyl chloride were added. The reaction mixture was stirred for 2 days at room temperature. Remove the solvent from the rotavapor and dissolve the residue in 50 ml of ethyl acetate and then wash with 1M HCl, 1M NaOH and distilled water. The organic phase was dried with sodium sulfate and the solvent removed under reduced pressure. The crude product was recrystallized from ethanol. Yield: 629 mg (19%)

Melting point: 142-144 ° C

IR (KBr, cm ^-1 ): 3415, 3197, 2915, 2224, 1648, 1422, 1329, 1156, 985, 786, 595 ¹ H-NMR (DMSO-d ₆ ); δ (ppm) 1.36-1.61 (m, 12H, CH ₂ -azepine); 2.09 (s, 3H, CO-CH ₃ );

2.81 and 2.91 (2t, 2H, J = 6:34, CH ₂ -isoquinoline _3); 3.10 (m, 2H, CH ₂ -isoquinoline ₄ ); 3.66 m, 2H, (CH2-isoquinoline); 4.26-4.41 (m, 2H, Ar-CH ₂ ), 7.35 and 7.52 (2d, 2H each, J ₂ , ₆ = 8.17, J ₃ , 5 = 8.17, Ar-H ₂ , 3.5, ₆ ); 7.31-7.80 (m, 3H, Ar-H (isoquinoline)); 9.40 (s, 1H NHSO ₂ ) Molecular Weight: calculated: 509 (C ₂₆ H ₃ and N ₅ O ₄ S); found: 510 (MH ⁺ )

EXAMPLE 5 (4 - {[2 - [(2-acetyl 1-1,2,3,4-tetrahydro-7-isoquinolin I) are Ifon yl] -1- (1 azepanylcarbonyl) hydrazino] methyl} phenyl) (imino) methanamine hydrochloride

-282-acetyl- N - (1-azepanylcarbonyl) - N '- (4-cyanobenzyl) -1,2,3,4-tetrahydro-7 isoquinolinsulfonohydrazide (600 mg, 1.18 mmol) was dissolved in 25 ml of anhydrous ethanol and HCI gas was introduced for 20 min. After complete administration, the solution was stirred for 4 hours at room temperature, then ammonium acetate (100.0 mg, 1.32 mmol) was added. The reaction mixture was left at room temperature for 2 days. We reintroduce HCI gas for 20 min. After 1 hour, the resulting ammonium chloride was filtered off and the ethanol was evaporated on a rotary evaporator. The product was poured into ether and filtered off by suction.

Yield: 312 mg (47%)

Melting point: 221-224 ° C

IR (KBr, cm ' ¹ ): 3417, 3287, 2946, 1636, 1423, 1334, 1158, 1091, 1036, 608 ¹ H-NMR (CDCl 3): δ (ppm) 1.22-1.74 (m, 12H, CH ₂ -azepine); 2.19 (s, 3H, CO-CH ₃ 2.91 (m, 2H, CH ₂ -isoquinoline ₃ ); 3.28 (m, 2H,, CH ₂ -isoquinoline ₄ ); 3.86 m, 2H,, CH ₂ isoquinolines); 4.35 (m, 2H, Ar-CH ₂ ), 7.21-7.72 (m, 7H, Ar-H); 7.96 (s, 1H NHSO ₂ ) Molecular weight C ₂₆ H35 N ₆ O ₄ S: calculated: 527; found: 528 (MH ⁺ )

EXAMPLE 6

7 - {[2- {4- [amino (imino) methyl] benzyl} -2- (1-azepanylcarbonyl) hydrazino] sulfonyl} -1,2,3,4tetrahydroisoquinoline dichloride (4 - {[2 - [(2- The acetates of 1-1,2,3,4-tetrahydro-7-isoquinol and 1-sulfonyl] -1- (1-azepanylcarbonyl) hydrazino] methyl} phenyl) (imino) methanamine hydrochloride (600 mg, 1.07 mmol) were dissolved in 15ml 4M HCI and heat for 2 hours at boiling. The water and the resulting acetic acid were evaporated on a rotary evaporator and the resulting colorless crystalline product was washed with ether.

Yield: 432 mg (72%)

Melting point: 226-228 ° C

IR (KBr, ^cm-1): 2932, 2598, 1698, 1669, 1424, 1382, 1235, 1155, 914, 736, 668 ¹ H-NMR (DMSO-d _6): δ (ppm) of 1:15 to 1:35 (w , 8H, CH ₂ - azepine ₃ , ₄ ', 5', ₆ '); 2.51 (m, 4H, CH ₂ -azepine ₂ ', ₇ '); 3.08 (m, 2H, CH ₂ -isoquinoline ₃ ); 3.21 (m, 2H, CH ₂ -isoquinoline ₄ ); 3.66 m,

-292Η, CH ₂ -isoquinoline); 4.26-4.41 (m, 2Η, Ar-CH ₂ ), 7.35 and 7.52 (2d, 2H each, J ₂ , ₆ = 8.17, J ₃ , 5 = 8.17, Ar-H ₂ , ₃ , 5, ₆ ); 7.31-7.80 (m, 3H, Ar-H (isoquinoline)); 9.40 (s, 1H NHSO ₂ ) Molecular Weight: calculated: 509 (C ₂₆ H ₃ and N ₅ O ₄ S); found: 510 (MH ⁺ )

EXAMPLE 7 N- (1-Azepanylcarbonyl) - N- (4-cyanobenzyl) -1- (2-naphthylsulfonyl) -2-pyrrolidinecarbohydrazide

2.20 g (7.21 mmol) of 1- (2-naphthylsulfonyl) proline were dissolved in 35 ml of dichloromethane and 815 mg (7.50 mmol) of ethyl chloroformate and 1.20 g (9.30 mmol) of Netyl-N, N-diisopropylamine were added with stirring. The reaction mixture was stirred for 1 hour at room temperature, then powdered 2- (1-azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazinium chloride (2.00 g,

6.48 mmol). After addition, it was stirred for 3 days at room temperature.

Remove the solvent from the rotavapor and dissolve the residue in 50 ml of ethyl acetate and then wash with 10% citric acid, 5% sol. NaHCO ₃ and distilled water. The organic phase was dried with sodium sulfate and the solvent removed under reduced pressure. The crude product is converted from acetone.

Yield: 0.82 g (23%)

Melting point: 158-159 ° C

IR (KBr, cm ' ¹ ) 3461, 2234, 1708, 1614, 1527, 1428, 1342, 1156, 1082, 1015 759, 662, 548

H-NMR (DMSO-d6): δ (ppm) 1.47 (s, 4H, CH ₂ ), 1.64 (s, 4H, CH ₂ ), 1.75 (m, 4H, CH ₂ ),

3.24 (m, 4H, CH ₂ ), 3.24 (m, 2H, CH ₂ ), 4.12 (m, 1H, CH). 4.47 (s, 2H, Ar-CH _? ), 7.57 (d, 2H, J = 8.29 Hz, Ar-H ³ ' ⁵ ), 7.71 (dqu, 2H, ^ = 7.16 Hz, J2 = 1.50 Hz, Ar-H ), 7.80 (d, 2H, J = 8.29 Hz, Ar-H ² ' ⁶ ), 7.84 (dd, 1H, Ji = 8.67 Hz, J2 = 1.89 Hz, Ar-H), 8.07 (d, 1H, J = 7.91 Hz, Ar-H), 8.15 (m, 2H, Ar-H), 8.47 (s, 1H, Ar-H), 10.29 (s, 1 H, NH)

Molecular Weight: calculated: 559 (C ₃₀ H ₃₃ N ₅ O ₄ S); found: 560 (MH ⁺ )

EXAMPLE 8

-3050

Amino {4 - [(1- (1-azepanylcarbonyl) -2 - {[1- (2-naphthylsulfonyl) -2-pyrrolidinyl] carbonyl} hydrazino) methyl] phenyl} methaniminium chloride

A / ^- (1-azepanylcarbonyl) - / V- (4-cyanobenzyl) -1- (2-naphthylsulfonyl) -2-pirolidinkarbohidrazid (754 mg, 1.35 mmol) of dissolved in 25 ml of anhydrous ethanol, and introducing HCl gas for 20 min. After complete administration, the solution was stirred for 4 hours at room temperature, then ammonium acetate (115.0 mg, 1.50 mmol) was added. The reaction mixture was left at room temperature for 2 days. We reintroduce HCI gas for 20 min. After 1 hour, the resulting ammonium chloride was filtered off and the ethanol was evaporated on a rotary evaporator. The product was poured into ether and filtered off by suction.

Yield: 320 mg (40%)

Melting point: 123-127 ° C

IR (KBr, ^cm-1): 3034, 1781, 1681, 1602, 1402, 1347, 1158, 1076, 1010, 823, 748, 660 ¹ H-NMR (DMSO-de): δ (ppm), 1:57 (m, 4H, CH2-azepine (4 ', 5'); 1.76 (m, 4H, CH 2 azepine 3 ', 6'); 2.03 (m, 4H, CH2-proline); 3.02 (m, 4H, CH2-azepin2 ', 7'); 3.32 (m, 2H, CH 2 proline); 3.63 (q, 1H, J = 3.63 Hz, CH2-proline); 4.96 (s, 2H, Ar-CH2), 7.52 (d, 2H, J = 8.18 Hz, Ar-H ³ ' ⁵ ), 7.68 (dqu, 2H, J, = 7.14 Hz, J2 = 1.67 Hz, Ar-H ), 7.81 (d, 2H, J = 8.18 Hz, Ar-H ² ' ⁶ ), 7.93 (dd, 1H, 3, = 8.59 Hz, J ₂ = 2.11 Hz, Ar-H), 8.12 (m, 3H. Ar-H),

8.49 (s, 1H, Ar-H), 9.41 (s, 4H, NH ₂ -C = NH ₂ ⁺ ) 9.59 (s, 1 H, NH)

Molecular Weight: Calculated: 599 (C30H37N5O4SCI); found: 577 ((M-HCl) H ⁺ )

EXAMPLE 9 N- (1-Azepanylcarbonyl) - N- (4-cyanobenzyl) -2- (2-naphthyloxy) acetohydrazide

750 mg (3.71 mmol) of 2- (2-naphthyloxy) acetic acid and 1.00 g (3.24 mmol) of 2- (1azepanylcarbonyl) -2- (4-cyanobenzyl) hydrazinium chloride were dissolved in 15 ml of dimethylformamide. With stirring, 500 mg (3.70 mmol) of 1-hydroxy-benzotriazole and 748 mg (3.90 mmol) of EDC are added. The reaction mixture was stirred at room temperature for 1 day.

-312> 4

Remove the solvent from the rotavapor and dissolve the residue in 50 ml of ethyl acetate and then wash with 10% citric acid, 5% sol. NaHCO ₃ and distilled water. The organic phase was dried with sodium sulfate and the solvent removed under reduced pressure. The crude product was purified by column chromatography (MF = CH ₂ Cl ₂ : MeOH 9: 1).

Yield: 963 mg (63%)

Melting point: 142-144 ° C

IR (KBr, cm ' ¹ ) 3228, 2925, 2854, 2227, 1630, 1509, 1426, 1216, 1120, 960, 838, 748, 547

H-NMR (CDCl ₃ ): δ (ppm) 1.55 (m, 4H, CH ₂ -azepine ₄ ', 5'); 1.75 (m, 4H, CH ₂ -azepine ₃ ', ₆ );

3.43 (t, 4H, J = 5.86 Hz, CH ₂ -azepine ₂ , ₇ ); 4.54 (s, 2H, CH ₂ ); 4.68 (s, 2H, Ar-CH ₂ ); 7.06 (s, 1H, NH); 7.34 (d, 2H, J = 8.40 Hz, Ar-H ^3.5 ); 7.39-7.58 (m, 3H, Ar-H); 7.47 (d, 2H, J = 8.40 Hz, Ar-H ^2.6 ); 7.68-7.84 (m, 3H, Ar-H); 8.14 (s, 1H, Ar-H)

Molecular Weight: Calculated: 456 (C ₂ 7H ₂₈ N ₄ O ₃ ); found: 457 (MH ⁺ )

EXAMPLE 10:

tert-butyl 2- {4 - [(acetylamino) methyl] benzyl} -2 - [(4-methyl-1-piperidinyl) carbon and 1] -1 hydrazinecarboxylate

1.26 g (4.39 mmol) of bis-trichloromethyl carbonate were dissolved in 20 ml of dichloromethane and the solution was purged with argon. 2.50 g (8.53 mmol) of tert-butyl 2- {4 [(acetylamino) methyl] benzyl} -1-hydrazinecarboxylate were dissolved in 30 ml of dichloromethane, 1.65 g (12.79 mmol) of diisopropylethylamine was added and the mixture was added dropwise to a solution of bis-trichloromethylcarbonate. While dripping, the reaction mixture was cooled in an ice bath. Then stir for another half hour at room temperature. 2.04 ml (25.60 mmol) of 4-methylpiperidine was added and stirred for another hour at room temperature. The reaction mixture was then extracted with 4 × 25 ml of 10% citric acid solution, 30 ml of saturated NaHCO ₃ solution, and washed with 30 ml of purified water and 20 ml of saturated NaCl solution. The organic phase is dried with Na ₂ SO ₄ . The solvent was evaporated off under reduced pressure and 10 ml of diethyl ether was added to the residue. A white precipitate precipitates and is filtered off by suction.

-322> 2Up: 2.10 g (59%)

Melting point: 124-126 ° C

IR (KBr, cm ' ¹ ) 3282, 2923, 2858, 1727, 1638, 1443, 1253, 1158, 1020, 978, 757, 611 H-NMR (CDCl ₃ ): δ (ppm) 0.95 (d, 2H , J = 6.40 Hz, pip-CH ₃ ); 1.10-1.25 (m, 2H, pipCH ₂ ); 1.44 (s, 9H, C (CH _{3) 3);} 1.60- 1.73 (m, 3H, pip-CH2, pip-CH); 2.03 (s, 3H, C0CH _3), 2.82 (m, 2H, pip-CH?): 3.92 (d, 2H, J = 13,19 Hz, pip-CH?); 4.42 (d, 4H, J = 5.65 Hz, CH ₂ -Ar-CH ₂ ), 5.89 (s, 1H, NH-CO); 6.27 (s, 1H, NH-COO); 7.27 (s, 4H, Ar-H) Molecular weight: calculated: 418 (C ₂₂ H ₃₄ N ₄ O ₄ ); found: 419 (MH ⁺ )

EXAMPLE 11:

2- {4 - [(acetylamino) methyl] benzyl} -2 - [(4-methyl-1-piperidine and 1) carbon and 1] hydrazinium chloride

2.65 g (6.33 mmol) of tert-butyl 2- {4 - [(acetylamino) methyl] benzyl} -2 - [(4-methyl-1-piperidinyl) carbonyl] -1-hydrazinecarboxylate were dissolved in acetic acid (20 ml) and we introduce HCI gas for half an hour. The acid is evaporated off under reduced pressure and the residue is taken up with diethyl ether. Using a glass rod, rub against the wall of the flask until a white powder is obtained. Meanwhile, double the diethyl ether and pour the new one.

Yield: 1.98 g (88%)

Melting point: 196-198 ° C

IR (KBr, cm ' ¹ ) 3419, 3256, 2926, 2685, 1689, 1552, 1434, 1235, 972, 728 H-NMR (CDCI ₃ ): δ (ppm) 0.97 (d, 2H, J = 6 , 03 Hz, pip-CH ₃ ); 1.19 (m, 2H, pip-CH2): 1.52-1.70 (m, 3H, pip-CH2, pip-CH); 2.07 (s, 3H, CO-CH ₃ ); 2.91 (t, 2H, J = 12.43 Hz, pip-CH?): 3.96 (d, 2H, J = 12.43 Hz, pip-CH?): 4.31 (d, 2H. J = 4.15 Hz, Ar-CH ₂ ); 4.61 (s, 2H, Ar-CH2); 7.26 (d, 2H, J = 7.91 Hz, Ar-H); 7.32 (d, 2H, J = 7.92 Hz, Ar-H); 7.99 (s, 1H, NH-CO)

Molecular Weight: calculated: 354.5 (0 ₁ 7Η ₂₇ Ν ₄ 0 ₂ ΟΙ); found: 319 ((M-HCl) H ⁺ , 100)

-33 EXAMPLE 12:

N- (4 - {[1 - [(4-methyl 1-1-pyridinyl) carbon and 1] -2- (1-naphthyl Isulfon and 1H-idrazino] methyl} benzyl) acetamide

1.98 g (5.59 mmol) of 2- {4 - [(acetylamino) methyl] benzyl} -2 - [(4-methyl-1piperidinyl) carbonyl] hydrazinium chloride and 1.39 g (6.13 mmol) of naphthalene-2-sulfonyl chloride were dissolved in dichloromethane (25 ml), 2.16 g (16.7 mmol) of diisopropylethylamine were added and stirred at room temperature for 3 days. It was extracted with 4 × 25 ml of 10% citric acid and washed with 30 ml of saturated NaHCO ₃ solution and the organic phase washed with 30 ml of purified water and 30 ml of saturated NaCl solution. We dried with Na ₂ SO ₄ . Dichloromethane was evaporated under reduced pressure to give a foamy solid, a pale brownish-yellow color.

Yield: 1.89 g (67%)

Melting point: 80-83 ° C

IR (KBr, cm ' ¹ ) 3285, 2925, 1656, 1546, 1430, 1338, 1165, 970, 750, 554 H-NMR (CDCI ₃ ): δ (ppm) 0.52 (d, 3H, J = 6 , 03 Hz, pip-CH ₃ ); 1.28 (m, 2H, pip-CHA 1.64 (m, 3H, pip-CH ₂ , pip-CH); 2.05 (s, 3H, CO-CH ₃ ); 2.32-2.71 (m, 2H, _pip-CH?); 3.72 (m, 2H, pip-CH _2); 4.32 (m, 2H, Ar-CH _2); 4.41 (d, 2H, J = 5 , 66 Hz, Ar-CH ₂ ); 5.72 (s, 1H, NH-CO); 7.16 (d, 2H, J = 8.29 Hz, Ar-H); 7.22 (d, 2H , J = 7.91 Hz, Ar-H); 7.46 (s, 1H, naphth-H); 7.62 (m, 2H, 2xnaphtha-H); 7.81 (dd, 1H, Τ = 8 , 67 Hz, J ₂ = 1.88 Hz, naphth-H);

7.94 (q, 3H, J = 7.66 Hz, 3xft-H); 8.45 (s, 1H, NH-SO ₂ )

Molecular Weight: Calculated: 508 (C ₂ 7H ₃₂ N ₄ O ₄ S); found: 509 (MH ⁺ )

EXAMPLE 13:

(4 - {[1 - [(4-methyl-1-pyridine I) carbon and 1] -2- (1-naphthyl Isulfonyl) hydrazo] methyl} phenyl) methanamine hydrochloride

1.89 g (3.72 mmol) of N- (4 - {[1 - [(4-methyl-1-piperidinyl) carbonyl] -2- (1 naphthylsulfonyl) hydrazino] methyl} benzyl) acetamide was dissolved in 30 ml of warmed

-34isopropyl alcohol (60 ° C). Add 30 ml of 4M HCl and heat for 5 hours at reflux temperature. Evaporate the solvent under reduced pressure. The product was purified by column chromatography using the stationary phase as silica gel, using dichloromethane: methanol (9: 1) as the mobile phase.

Yield: 550 mg (29%)

Melting point: 156-160 ° C

IR (KBr, cm ' ¹ ) 3285, 2925, 1656, 1546, 1430, 1338, 1165, 970, 750, 554

H-NMR (CDCI _3): δ (ppm) 0.17 (m, 2H, _pip-CH?), 0.44 (d, 3H, J = 6.03 Hz, pip-CH _3),

1.22 (d, 2H, J = 10.54 Hz, pip-CH ₂ , pip-CH), 2.27-2.74 (m, 2H, pip-CH ₂ ), 3.53 (s, 2H , pip-CH?), 3.97 (s, 2H, Ar-CH?) 4.32 (m, 2H, Ar-CH _? ), 7.22 (d, 2H, J = 8.29 Hz, Ar -H), 7.42 (d, 2H, J = 8.29 Hz, Ar-H), 7.64-7.75 (m, 3H, 3x naphtha-H), 7.77 (dd, 1H, N). = 8.67 Hz, J ₂ = 1.88 Hz, naphtha-H), 7.81-8.00 (m, 1H, naphtha-H), 8.02-8.18 (m, 2H, 2x naphtha- H), 8.46 (d, 1H, J = 1.88 Hz, NH-SO ₂ )

Molecular Weight: Calculated: 502.5 (C ₂ 5H ₃₁ N ₄ O ₃ SCI); found: 467 ((M-HCl) H ⁺ )

EXAMPLE 14:

tert-butyl 2- {4 - [(acetylamino) methyl] benzyl} -2- (4-morpholinylcarbonyl) -1hydrazincarboxylate

1.26 g (4.39 mmol) of bis-trichloromethyl carbonate were dissolved in 20 ml of dichloromethane and the solution was purged with argon. 2.50 g (8.53 mmol) of tert-butyl 2- {4 [(acetylamino) methyl] benzyl} -1-hydrazinecarboxylate were dissolved in 30 ml of dichloromethane, 1.65 g (12.79 mmol) of diisopropylethylamine was added and the mixture was added dropwise to a solution of bis-trichloromethylcarbonate. While dripping, the reaction mixture was cooled in an ice bath. Then stir for another half hour at room temperature. 2.23 ml (25.60 mmol) of morpholine were added and stirred for another hour at room temperature. The reaction mixture was then extracted with 4 * 25 ml of 10% citric acid solution, 30 ml of saturated NaHCO ₃ solution, and washed with 30 ml of purified water and 20 ml of saturated NaCl solution.

-35 The organic phase is dried with Na ₂ SO ₄ . The solvent was evaporated off under reduced pressure and 10 ml of diethyl ether was added to the residue. A white precipitate precipitates and is filtered off by suction. Yield: 1.81 g (52%)

Melting point: 125-128 ° C

IR (KBr, ^cm-1) 3308, 2980, 2857, 1728, 1641, and 1430 1289 1114 1025 873, 746, 604 H-NMR (CDCl): δ (ppm) 1.46 (s, 9H, C (CH ₃ ) ₃ ), 2.03 (s, 3H, CO-CH ₃ ), 3.44 (t, 4H, J = 4.71 Hz, 2xmorph-CH ₂ ), 3.68 (t, 4H , J = 4.71 Hz, 2xmorph-CH ₂ ), 4.43 (d, 2H, J = 5.65 Hz, Ar-CH?), 4.49 (s, 2H, Ar-CH?), 5 , 84 (d, 1H, NH-CO), 6.27 (s, 1H, NH-COO), 7.27 (s, 4H, Ar-H).

Molecular Weight: calculated: 406 (0 ₂₀ Η ₃ οΝ ₄ 0 ₅ ); found: 407 (MH ⁺ )

EXAMPLE 15:

2- {4 - [(acetylamino) methyl] benzyl} -2- (4-morpholinylcarbonyl) hydrazinium chloride

1.81 g (4.46 mmol) of tert-butyl 2- {4 - [(acetylamino) methyl] benzyl} -2- (4-morpholinylcarbonyl) 1-hydrazinecarboxylate was dissolved in acetic acid (20 ml) and HCI gas was introduced. hours. The acid is evaporated off under reduced pressure and the residue is taken up with diethyl ether. Using a glass rod, rub against the wall of the flask until a white powder is obtained. Meanwhile, double the diethyl ether and pour the new one.

Yield: 0.92 g (60%)

Melting point: 209-211 ° C

IR (KBr, cm ' ¹ ) 3427, 1685, 1560, 1432, 1274, 1112, 1022, 892, 571

H-NMR (CDCI ₃ ): δ (ppm) 1.88 (s, 3H, CO-CH ₃ ), 3.46 (d, 4H, J = 4.90 Hz, 2xmorph-CH _? ), 3.61 (d, 4H, J = 4.90 Hz, 2xmorph-CH?). 4.25 (d, 2H, J = 6.03 Hz, Ar-CH?). 4.50 (s, 2H, Ar-CH ₂ ), 7.28 (s, 4H, Ar-H), 8.43 (s, NH-CO)

Molecular Weight: calculated: 342.5 (Ci ₅ H ₂₃ N ₄ O ₃ CI); found: 307 ((M-HCl) H ⁺ , 100)

EXAMPLE 16:

-3656

N- (4 - {[1- (4-morpholinyl and Icarbonyl I) -2- (1-naphthyl Isu Ifon and l) hydrazes ηο] methyl} benzyl) acetamide

0.92 g (2.69 mmol) of 2- {4 - [(acetylamino) methyl] benzyl} -2- (4morpholinylcarbonyl) hydrazinium chloride) and 0.67 g (2.95 mmol) of naphthalene-2 sulfonyl chloride were dissolved in dichloromethane (25 ml), 1.04 g (8.06 mmol) of diisopropylethylamine were added and stirred at room temperature for 3 days. It was extracted with 4 * 25 ml of 10% citric acid and washed with 30 ml of saturated NaHCO ₃ solution and the organic phase washed with 30 ml of purified water and 30 ml of saturated NaCl solution. We dried with Na ₂ SO ₄ . Dichloromethane was evaporated under reduced pressure to give a foamy solid, a pale brownish-yellow color.

Yield: 0.66 g (50%)

Melting point: 84-88 ° C

IR (KBr, ^cm-1) 3426, 2856, 1655, 1420, 1340, 1274, 1166, 1114, 1024, 752, 547 H-NMR (CDCI _3): δ (ppm) 2.05 (s, 3H, CH ₃ ), 3.14 (s, 4H, 2 * morph-CH ₂ ), 3.33-3.40 (m, 2H, morph-CH ₂ ), 3.58-3.81 (m, 2H, morph -CH _? ), 4.30 (m, 2H, Ar-CH ₂ ), 4.41 (d, 2H, J = 5.65 Hz, Ar-CHA 5.77 (s, 1H, NH-CO), 7.19 (m, 4H, Ar-H), 7.38 (s, 1H, naphth-H),

7.68 (m, 2H, 2xnaft-H), 7.81 (dd, J, = 8.67 Hz, J ₂ = 1.86 Hz, naphtha-H), 7.95 (m, 3H, 3xnaft- H), 8.44 (s, NH-SO ₂ )

Molecular Weight: Calculated: 496 (C ₂ H ₂₈ N ₄ O ₅ S); found: 497 (MH ⁺ )

EXAMPLE 17:

(4 - {[1- (4-morpholinylcarbonyl) -2- (1-naphthylsulfonyl) hydrazino] methyl} phenyl) methanamine hydrochloride

0.66 g (1.33 mmol) of N- (4 - {[1- (4-morpholinylcarbonyl) -2- (1-naphthylsulfonyl) hydrazino] methyl} benzyl) acetamide was dissolved in 30 ml of heated isopropyl alcohol (60 ° C). ). Add 30 ml of 4M HCl and heat for 5 hours at reflux temperature. Evaporate the solvent under reduced pressure. The product is purified by column chromatography, where appropriate

-37 'to stationary silica gel phase, using dichloromethane: methanol (9: 1) as the mobile phase.

Yield: 340 mg (52%)

Melting point: 124-126 ° C

IR (KBr, cm ' ¹ ) 3411, 16662, 1504, 1459, 1419, 1335, 12272, 1211, 1166, 1113, 1067, 1023, 830, 754, 688, 544

H-NMR (CDCl3); δ (ppm) 2.96 (m, 2H, morph-CHA 3.11 (m, 6H, 3 * morph-CHA 3.97 (d, 2H, J = 5.66 Hz, Ar-CH?), 4 , 17-4.52 (m, 2H, Ar-CH _? ), 7.22 (d, 2H, J = 7.53 Hz, Ar-H), 7.41 (d, 2H, J = 7.92 Hz, Ar-H), 7.82-7.66 (m, 3H, 3 * naphth-H), 7.99-7.84 (m, 1H, naphtH), 8.03-8.21 (m , 3H, 3 * naphth-H), 8.47 (s, 1H, NH-SO ₂ )

Molecular Weight: Calculated: 490.5 (C25H31N4O3SCI); found: 455 ((M-HCl) H ⁺ )

Lek, a pharmaceutical factory

University of Ljubljana Faculty of Pharmacy

Claims (8)

PATENT APPLICATIONS Compounds of formula I R ¹ O) R ³ X N ^ R ² NH in which they mean R ^{1 is a} residue of formula h ₂ n ^ nr < , NH, Cl where R ⁴ = H, alkyl (C 1 -C ₃ ), OH, O-alkyl (C 1 -C ₃ ), NH ₂ ; R is the residue of the formula R ⁵ R ⁶ —N R ⁷ -N -N ESD ¹¹ V-X R ⁸ X \ _R9 where R ⁵ = H, alkyl (CrCa), COOR ¹⁰ , R ⁶ = H, alkyl (C 1 -C ₃ ), COOR ¹⁰ , R ⁷ = H, alkyl (θ! -Θ ₃ ), COOR ¹⁰ , R ¹⁰ = H, alkyl (C 1 -C ₃ ), R ⁸ = H, alkyl (C ₁ -C ₆ ), cycloalkyl (C ₃ -C ₆ ), R ⁹ = H, alkyl (C 1 -C 3), cycloalkyl (C ₃ -C ₆ ), R ¹¹ = H, alkyl (C 1 -C ₃ ) , benzyl, X = ch ₂ , o, s, -39i *) Y = NR ¹² , Ο, S, R ¹² = H, COCH ₃ , alkyl (C 1 -C ₃ ); R ^{3 is a} residue of the formula ° '' P Ά h, c. o, o o O, .o ca ..... οτ -'- οο · ' o. o or a radical of the formula in the case where R ¹ =, νηΪοι and their pharmaceutically acceptable salts. Compounds of formula I according to claim 1 for use as therapeutically active substances. A process for the preparation of compounds of the formula I according to claim 1, characterized in that H ₂ N 4 NR 4 a) in the case where R ¹ = ' 4-Cyanobenzaldehyde of Formula (II) -40CN (H) co CHO is converted with a BOC-carbazate of formula (lil) (III) into a compound of formula (IV) CN AT N ' I -N H (IV) which is then converted to compound (V) by a catalytic hydrogenation reduction process CN (V) HN O ^{ΝΗ 1} o J reacting with triphosgene and an amine of formula (VI) HN R ⁷ (VI) wherein R ⁵ , R ⁶ and R ^{7 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (VII or Vlyl), TJ (VII) (Vlil) -41 or with triphosgene and an amine of formula (IX) Γ HN. ^{R 9} (IX) wherein R ⁸ and R ^{9 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (X), COOR ¹¹ HN (X) where R ^{11 has the} same meaning as in formula (I), to compound (XI) where R ^{2 has the} same meaning as in formula (I); and then the BOC protecting group in compound (XI) is removed by HCl (g) in HOAc at room temperature to give compound (XII), where R ^{2 has the} same meaning as in formula (I) which reacts with aromatic sulfonyl chloride or naphthylsulfonyl amino acid with activated carboxyl group to compound (XIII) -4242CN wherein R ² and R ^{3 have the} same meaning as in formula (I), which is then reacted with hydroxylamine in anhydrous ethanol to compound of formula (XIV) R ¹ I NH ^H 2 ^N \ ^> ^NR4 where R ¹ = | , R ² and R ³ have the same meaning as in formula (I), and R ⁴ represents OH; or is metabolised by introducing gaseous HCI into methanolic solution by the addition of ammonium acetate and reintroducing HCI into compound (XV), R ¹ r O where h ₂ n ^ nr < R ¹ = 'R ² and R ³ have the same meaning as in formula (I), and R ⁴ represents hydrogen; and in the case -43b) when R ¹ = NH, Cl te 4-Cyanbenzaldehyde of Formula (II) CN CHO is converted with ethylene glycol in the presence of 4-toluenesulfonic acid to compound (XVI) CN which is then reduced with lithium aluminum hydride to a compound of formula (XVII) ch ₂ nh ₂ ό 1 (XVII) ° \ _ / ° reacting with acetanhydride to compound (XVIII) ch ₂ nhcoch ₃ ά 1 (XVIII) C> Ό which is converted with 90% methanoic acid to compound (XIX) cho (xix) which is converted with BOC-carbazate of formula (III) -44 (III) JA to a compound of formula (XX) ch ₂ nhcoch ₃ >(XX)> fY ^{H 1} o which is converted into a compound (XXI) ch ₂ nhcoch ₃ (xxi) by a catalytic hydrogenation reduction process R ⁵ R ^with reacting with triphosgene and an amine of formula (VI) HN R ⁷ (VI) wherein R ⁵ , R ⁶ and R ^{7 have the} same meaning as in formula (I), or with triphosgene and amine of formula (VII or Vlil), ^H vJ (Vil) (Vlil), or triphosgene and amine with the formula (IX) Γ HN, ^{R 9} (IX) wherein R ⁸ and R ^{9 have the} same meaning as in formula (I), or with triphosgene and an amine of formula (X), -45 ^ b; oor ¹¹ HNJ ( ^χ ) where R ^{11 has the} same meaning as in formula (I), to compound (XXII) where R ^{2 has the} same meaning as in formula (I); then the BOC protecting group in compound (XXII) is removed with HCl (g) in AcOH at room temperature to give compound (XXIII), ch ₂ nhcoch ₃ (XXIII) nh; _c r where R ^{2 has the} same meaning as in formula (I) which reacts with aromatic sulfonyl chloride to compound (XXIV) ch ₂ nhcoch ₃ o (XXIV) R2 I, NH R * where R ² and R ^{3 have the} same meaning as in formula (I) and are then digested by boiling in 5M HCl to compound (XXV) R ' O (XXV) -46 where R ¹ = and R ² and R ³ have the same meaning as in formula (I). Use of compounds of formula I for the preparation of medicaments having anticoagulant activity. 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 for use as anticoagulants. Pharmaceutical preparations according to claim 5 for use in the inhibition of thrombin in the blood of man and other mammals. Pharmaceutical compositions according to claim 5, characterized in that they are used to inhibit the formation of fibrin and thrombus in the blood of humans and other mammals. Lek, a pharmaceutical and chemical factory, d.d. University of Ljubljana Faculty of Pharmacy