Sulfur-containing compounds, methods for their preparing and pharmaceutical preparations based on thereof

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new sulfur-containing compounds of the formula (I):

their pharmaceutically acceptable salts or solvates, or salt solvates wherein R1 represents (C1-C6)-alkyl, cycloalkyl, aryl, aliphatic or aromatic heterocyclyl substituted with one more basic group, such as amino-, amidino- and/or guanidine-group; R2 represents hydrogen atom (H), alkyl, alkylthio-, alkoxy- or cycloalkyl group; R3 represents COOR5, SO(OR5), SOR5 and others; R4 represents hydrogen atom (H) or (C1-C6)-alkyl; R6 represents hydrogen atom (H); X represents C(Z)2 or NR6CO; Y represents C(Z)2; Z represents hydrogen atom (H), (C1-C6)-alkyl, aryl or cycloalkyl. Indicated compounds inhibit activity of carboxypeptidase U and can be used for prophylaxis and treatment of diseases associated with carboxypeptidase U.

EFFECT: improved preparing method, valuable biochemical and medicinal properties of compounds.

14 cl, 36 ex

 

The scope of the invention

The present invention relates to new compounds and their pharmaceutically acceptable salts, which inhibit the basic carboxypeptidase, more specifically the carboxypeptidase U and thus can be used in the prevention and treatment of diseases in which inhibition of carboxypeptidase U is beneficial. In other aspects the invention relates to compounds according to the invention for use in therapy; to methods of producing these new compounds; to pharmaceutical compositions containing at least one compound according to the invention or its pharmaceutically acceptable salt as an active ingredient; and for use of the active compounds in the manufacture of drugs for medical use, as specified above.

Background of the invention

Fibrinolysis is the result of a series of enzymatic reactions leading to the degradation of fibrin by plasmin. Activation of plasminogen is a Central process in fibrinolysis. Cleavage of plasminogen with getting plasmin carry out the activators of plasminogen - plasminogen activator tissue type (t-PA) or plasminogen activator urokinase type (u-PA). During the initial cleavage of fibrin by plasmin formed carboxykinase lysine residues, which serve as high-affinity sites is wyzwania plasminogen. As the plasminogen bound to fibrin, is activated to plasmin is much easier than free plasminogen, this mechanism provides for the regulation of fibrinolysis by type of positive feedback.

One of the endogenous inhibitors of fibrinolysis is the carboxypeptidase U (CPU). The CPU is also known as plasma carboxypeptidase, active inhibitor of thrombin-activatable fibrinolysis (TAFIa), carboxypeptidase R and induced carboxypeptidase activity. CPU is formed during coagulation and fibrinolysis from its predecessor, CPU under the action of proteolytic enzymes such as thrombin, thrombin-thrombomodulin complex or plasmin. CPU cleaves basic amino acids at the carboxyl end of fibrin fragments. The loss of the carboxy-terminal lysine and, as a result, the lysine binding sites of plasminogen then contributes to the inhibition of fibrinolysis.

It is expected that inhibition of the loss of lysine binding sites of plasminogen and, as a result, increase the rate of formation of plasmin, effective inhibitors of carboxypeptidase U will facilitate fibrinolysis.

2-Mercaptomethyl-3-guanidinopropionic acid is known as an inhibitor of carboxypeptidase N. Recently, it was shown that this compound inhibits CPU [Hendriks, D. et al., Biochimica et Biophysica Acta, 1034 (1990) 86-92.

Guanidinoacetate acid is known as an inhibitor of carboxypeptidase N. Recently, it was shown that this compound inhibits CPU [Eaton, D.L. et al., The Journal of Biological Chemistry, 266 (1991) 21833-21838].

Description of the invention

It has been unexpectedly discovered that compounds of the formula I are particularly effective as inhibitors of carboxypeptidase U and, therefore, are suitable as medicaments for the treatment or prevention of conditions in which inhibition of carboxypeptidase U is beneficial.

Thus, in one aspect this invention relates to compounds of General formula I,

or their pharmaceutically acceptable salts or solvate, or solvate of such a salt, where

R1represents a

C1-C6alkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one nitrogen atom;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

or aryl substituted by one or more than one basic group such as amino, amidino and/or guanidino;

R2represents H, acyl, acylamino, alkyl, allylcarbamate, alkylthio, alkoxy, aroyl, aroylamino, aryloxy, aaltio, amidino, amino, aryl, carbarnoyl, carboxy, cyano, cycloalkyl, formyl, guanidino, halogen, heterocyclyl, hydroxy, oxo, nitro, thiol, group, Z2N-CO-O-, ZO-CO-NZ - or Z2N-CO-NZ-;

R3represents COOR5, SO(OR5), SO3R5P=O(or SIG5)2B(OR5)2P=OR5(OR5), or tetrazol, or any isaster carboxylic acids;

R4represents SH, S-CO-C1-C6alkyl or S-aryl;

R5represents H, C1-C6alkyl or aryl;

R6represents N or C1-C6alkyl;

X represents O, S, SO, SO2C(Z)2N(Z), NR6SO2, SO2NR6, NR6CO or CONR6;

Y represents C(Z)2;

Z independently represents H, C1-C6alkyl, aryl, cycloalkyl or heterocyclyl.

Preferred compounds of the present invention are the compounds of formula I or their pharmaceutically acceptable salt or solvate, or a solvate of such a salt, where

R1represents a

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl that contains at the ore one nitrogen atom;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

or aryl substituted by one or more than one basic group such as amino, amidino and/or guanidino;

R2represents H, acyl, acylamino, alkyl, allylcarbamate, alkylthio, alkoxy, aroyl, aroylamino, aryloxy, aaltio, amidino, amino, aryl, carbarnoyl, carboxy, cyano, cycloalkyl, formyl, guanidino, halogen, heterocyclyl, hydroxy, oxo, nitro, thiol, group, Z2N-CO-O-, ZO-CO-NZ - or Z2N-CO-NZ-;

R3represents COOR5;

R4represents SH, S-CO-C1-C6alkyl or S-CO-aryl;

R5represents H, C1-C6alkyl or aryl;

R6represents N or C1-C6alkyl;

X represents O, S, SO, SO2C(Z)2N(Z), NR6SO2, SO2NR6or CONR6;

Y represents C(Z)2;

Z independently represents H, C1-C6alkyl, aryl, cycloalkyl or heterocyclyl.

More preferred compounds of the present invention are the compounds of formula I or their pharmaceutically acceptable salt or solvate, or a solvate of such a salt, where

R1represents a

recloak is l, substituted one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one nitrogen atom;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

R2represents H, C1-C3alkyl, amino, halogen or hydroxy;

R3represents COOR5;

R4represents SH, S-CO-C1-C6alkyl or S-CO-aryl;

R5represents H, C1-C6alkyl or aryl;

X represents C(Z)2;

Y represents C(Z)2;

Z independently represents N or C1-C6alkyl.

Even more preferred compounds according to the present invention are the compounds of formula I or their pharmaceutically acceptable salt or solvate, or a solvate of such a salt, where

R1represents a

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one nitrogen atom;

R2represents H, F or C1alkyl;

R3represents COOR5;

R4represents SH, S-CO-C1-C6alkhalili S-CO-aryl;

R5represents H, C1-C6alkyl or aryl;

X represents C(Z)2;

Y represents C(Z)2;

Z independently represents N or C1-C6alkyl.

The most preferred compounds according to the present invention are the compounds of formula I or their pharmaceutically acceptable salt or solvate, or a solvate of such a salt, where

R1is cyclopentyl, pyridyl, pyrimidinyl, piperidinyl or thiazolyl;

R2represents H, F or C1alkyl;

R3represents COOR5;

R4is a SH;

R5represents N;

X represents CHZ;

Y represents CHZ;

Z independently represents N or C1-C6alkyl.

In the description of the invention and the attached formula used the following definitions of terms.

The term "core group" means the principal group, with the conjugated acid of the specified core group has a pKa from about -5 to about 25, preferably from 1 to 15.

The term "ISOStAR carboxylic acid" means an acid group having a pKa from about -5 to about 25, preferably from 1 to 15.

The term "C1-C6alkyl" means a straight or branched, saturated or nanosys is nnow, substituted or unsubstituted alkyl group having from 1 to 6 carbon atoms in the chain, and this alkyl group may be interrupted by one or more than one heteroatom selected from O, N or S. Examples of the specified alkyl include, without limitation, methyl, ethyl, ethynyl, ethinyl, n-propyl, isopropyl, propenyl, Isopropenyl, PROPYNYL, n-butyl, isobutyl, sec-butyl, tert-butyl, butenyl, Isobutanol, butenyl and pentyl and hexyl straight or branched chain.

The term "C1-C3alkyl" means a straight or branched, saturated or unsaturated, substituted or unsubstituted alkyl group having from 1 to 3 carbon atoms in the chain, and the alkyl group may be interrupted by one or more than one heteroatom selected from O, N or S. Examples of the specified alkyl include, without limitation, methyl, ethyl, ethynyl, ethinyl, n-propyl, isopropyl, propenyl, Isopropenyl, PROPYNYL.

The term "C1alkyl" means a substituted or unsubstituted alkyl group having 1 carbon atom. An example of the specified alkyl includes, without limitation, methyl.

The term "C1-C6alkoxy" means alkyl-O-group, where C1-C6alkyl such as defined above.

The term "C1-C3alkoxy" means alkyl-O-group, where C1-C3alkyl such as is designated above.

The term "heterocyclyl" means substituted or unsubstituted, 4-10-membered monocyclic or cyclical ring system in which one or more than one atom in the ring or in the ring is an element other than carbon, for example nitrogen, oxygen or sulfur, in particular 4-, 5 - or 6-membered aromatic or aliphatic heterocyclic group, and includes, without limitation, azetidinone, furan, thiophene, pyrrole, pyrrolidone, pyrolidine, dioxolane, oxathiolane, oxazolinone, oxazoline, thiazole, imidazole, imidazoline, imidazolidinone, pyrazol, pyrazolinone, pyrazolidinone, isoxazoline, isothiazol, oxadiazoline, hursanboy, triazole, thiadiazole, pornowww, pyridine, piperidine, dioxane, morpholino, diciannove, occationaly, thiomorpholine, pyridazinone, pyrimidine, pyrazinone, pieperazinove, triazine, thiadiazine, diazinone, isoindoline, isoindolines, indole, indolinone, naphthyridinone group and that obviously includes all isomers of the above groups. Obviously, for example, the term "azetidinol" includes 2 - and 3-isomers, and the term "pyridyl" and "piperidinyl" includes 2-, 3 - and 4-isomers.

The term "cycloalkyl" means a saturated or unsaturated, substituted or unsubstituted, neurom the political ring, consisting of 3, 4, 5, 6 or 7 carbon atoms, and includes, without limitation, group cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclopentadienyl, cyclohexadienyl or cycloheptadiene.

The term "halogen" includes groups with fluorescent, chloro, bromo and iodide.

The term "aryl" means substituted or unsubstituted With6-C14aromatic hydrocarbon and includes, without limitation, phenyl, naphthyl, indenyl, anthracene, phenanthrene and fluorene.

The term "aryloxy" means aryl-O-group, where the aryl as defined above.

The term "acyl" means an alkyl-CO-group, where alkyl such as defined above.

The term "aroyl" means an aryl-CO-group, where the aryl as defined above.

The term "alkylthio" means alkyl-S-group, where alkyl such as defined above.

The term "aristeo" means an aryl-S-group, where the aryl as defined above.

The term "aroylamino" means aroyl-N(Z)-group, where aroyl and Z are such as defined above.

The term "acylamino" means acyl-N(Z)-group, where the acyl and Z are such as defined above.

The term "carbarnoyl" means H2N-CO-rpynny.

The term "allylcarbamate" means Z2N-CO-rpynny, where Z is as defined above.

The term "substituted" means a group "C1alkyl", "C1 -C3alkyl, C1-C6alkyl, cycloalkyl", "heterocyclyl" or "aryl"as defined above which is substituted one or more than one acyl, acylamino, alkyl, alkylcarboxylic, alkylthio, alkoxy, aroline, aroylamino, aryloxy, aaltio, amidino, amino, aryl, carbamoyl, carboxy, cyano, cycloalkyl, formyl, guanidino, halogen, heterocyclyl, hydroxy, oxo, nitro, Tilney, tio, Z2N-CO-O-, ZO-CO-NZ - or Z2N-CO-NZ-group.

In addition, the compounds of formula I, where R4represents mercapto, can be in the form of a dimer linked via-S-S-bond, which is also included in the invention.

As pure enantiomers and racemic mixtures and unequal mixtures of two enantiomers are included in the scope of the present invention. It is obvious that all possible diastereomeric forms are included in the scope of the present invention. Also included in this invention derivatives of compounds of formula I with the biological function of compounds of formula I, such as prodrugs.

Depending on the conditions of the method of obtaining the compounds of formula I are produced either in a neutral or salt form or in the form of MES, such as a hydrate, and all of them are included in the scope of the present invention.

Getting

According to the present invention is also suggested ways And With the receiving connection is s the General formula I.

Method And

Method And obtain the compounds of General formula I, where R1, R2, R3, R4and Y are such as defined above, R4represents N and X represents a C(Z)2includes the following stages:

a) compounds of General formula II,

where R1such as defined for formula I, X represents a C(Z)2that are either available commercially or can be produced using known methods, can be converted into a compound of General formula III,

where L is a suitable leaving group such as chloro, bromo, iodo, triflate or tosyl, under standard conditions using a suitable reagent, such as h3/GVS4, TosCl/pyridine or (CF3SO2)2O/Thea;

b) compounds of General formula III can then be subjected to interaction with compounds of General formula IV,

where R2and R3such as defined for formula I, which are either available commercially or can be produced using known methods, in the presence of a suitable base, such as2CO3or NaH, in standard conditions to form compounds of General formula V

C) compounds of General fo the mules V, where R1and R3such as defined for formula I, X represents C(Z)2and R2represents H, can then be converted into compounds of General formula VI

by treatment with formaldehyde in the presence of a suitable base such as Et2NH, in standard conditions;

g) compounds of General formula VI can also be obtained by treating compounds of General formula VII,

where R3and R5such as defined for formula I with an alkylating agent of General formula III in the presence of a suitable base, such as LDA or NaH, in standard conditions to form compounds of General formula VIII

d) compounds of General formula VIII can then be subjected to interaction with the corresponding aldehyde or ketone OC(Z)2in the presence of a suitable base, such as KOtBu, LDA or NaH, in standard conditions to obtain compounds of General formula VI

e) compounds of General formula VI can then be subjected to interaction with compounds of General formula IX,

where R5is a suitable protecting group, such as Ac, Bz, RMV or EAP, either alone or in the presence of a suitable base, such the AK NaOMe, NaH or triethylamine, or, alternatively, in the presence of a free radical initiator, such as AIBN, under standard conditions to obtain compounds of General formula I, where R1, R2, R4and Y are such as defined for formula I, a R2represents H, and X is C(Z)2.

Method B

Method B to obtain the compounds of General formula I, where R1, R2, R3and R4such as defined in formula I, Y is CH2, a X represents O, S, SO, SO2C(Z)2or N(Z), includes the following stages:

a) compound of General formula X,

where R1such as defined for formula I and X represents O, S or N(Z), is subjected to the interaction with an alkylating agent of General formula XI,

where R2and R3such as defined for formula I, L is a suitable leaving group such as chloro, bromo, iodo, triflate or toiletry group, under standard conditions, using suitable reagents, such as NaH, Hell2CO3or Bu4NHSO4/NaOH, prior to the formation of compounds of General formula XII

b) compounds of General formula XII can then be subjected to the interaction with carbon dioxide in the presence of a suitable base such as LDA or KHMDS, under standard conditions to form compounds of General formula XIII

C) compounds of General formula XIII can then be subjected to interaction with alkylchlorosilanes, such as lC, in the presence of a base, such as triethylamine, and then the resulting mixed anhydride can be recovered with a suitable regenerating agent such as NaBH4in standard conditions with the formation of compounds of General formula XIV

g) compounds of General formula XIV can then be subjected to interaction with the compound of General formula IX

where R5is a suitable protecting group, such as AC or Bz, in the presence of a suitable reagent, such as PPh3/DIAD, under standard conditions to obtain compounds of General formula I, where R1, R2, R3and R4such as defined above, Y is CH2and X represents O, S, C(Z)2or N(Z);

d) compounds of General formula I, where R1, R2, R3and Y are such as defined above, and X is S, can then be subjected to interaction with a suitable oxidizing reagent, such as MSRA, under standard conditions to obtain compounds of General formula I, where R1, R2, R3and R4and Y such as is predelino above, but X is SO or SO2.

Method In

The method of obtaining compounds of General formula I, where R1, R2, R3and R4and Y are such as defined above, and X represents NR6CO, CONR6, SO2NR6or NR6SO2includes the following stages:

a) compound of General formula XV

where R2, R3and R6and Y are such as defined for formula I, a R5is a suitable protecting group, such as Ac, Bz, PMB, or EAP, is subjected to the interaction with the compound of General formula XVI

where R1such as defined for formula I, X represents a COOH or SO2Cl, in the presence of suitable linking agents such as PyBOP/DIPEA, DCC/HOBt, EDC/TEA/DMAP or pyridine, under standard conditions to obtain compounds of General formula I, where R1, R2, R3and R4and Y are such as defined above, and X is NR6CO or NR6SO2;

b) the compound of General formula XVII

where R2, R3and Y are such as defined for formula I, X represents COOH or SO2Cl, and R5is a suitable protecting group, such as AC, Bz, PMB, or EAP, is subjected to the interaction with the compound of General formula XVIII

where R6 such as defined for formula I, in the presence of suitable coupling reagents such as PyBOP/DIPEA, DCC/HOBt, EDC/TEA/DMAP or pyridine, under standard conditions to obtain compounds of General formula I, where R1, R2, R3and R4and Y are such as defined above, and X represents CONR6or SO2NR6.

For specialists in the art it is obvious that the methods described above, the functional groups of intermediate compounds may need to be protected with suitable protective groups.

Functional group, which is desirable to protect include hydroxy, amino, mercapto and carboxyl. Suitable protective groups for hydroxy include trialkylsilyl or diarylethylenes (for example, tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl and benzyl. Suitable protective groups for amino, amidino or guanidino include tert-butyloxycarbonyl and benzyloxycarbonyl. Suitable protective groups for mercapto include CO1-6alkyl, para-methoxybenzyl, trityl. Suitable protective groups for carboxyl include C1-6alkilany and benzyl esters.

The protective group can be removed by methods known to experts in the art and which are described below.

Some protected derivatives of the compounds is of the formula I, which can be obtained up to the last stage of removing protection from the formation of compounds of formula I are new.

The use of protective groups described in "Protective Groups in Organic Synthesis", 2nd edition, T.W.Greene & P.G.M.Wutz, Wiley-lnterscience (1991). The protective group may also be a resin, such as Wang resin or 2-horticulturally resin.

For specialists in this area it is obvious that although such protected derivatives of compounds of formula I may not possess pharmacological activity, they can be administered parenterally or orally, and thereafter they are metabolized in the body to form compounds of the present invention which are pharmacologically active. Such derivatives can therefore be described as prodrugs. All prodrugs of the compounds of formula I are included in the scope of this invention.

It is also clear that all polymorphs, amorphous forms, anhydrate, a hydrate, a solvate of the compounds of the present invention are included in the scope of this invention.

Pharmaceutical

In another aspect this invention relates to pharmaceutical compositions containing at least one compound of the present invention or its pharmaceutically acceptable salt as an active ingredient.

For clinical use of the compounds according to the present which the invention is administered in the form of pharmaceutical preparations for oral, intravenous, subcutaneous, tracheal, bronchial, intranasal, pulmonary, transdermal, buccal, rectal, parenteral or other techniques. The pharmaceutical preparation contains the compound of the present invention in combination with one or more pharmaceutically acceptable ingredient. The carrier may be in the form of solid, semi-solid or liquid diluent or capsules. Such pharmaceutical preparations are another object of the present invention. Usually the number of active compounds is 0.1-95% by weight of the drug.

During the preparation of pharmaceutical preparations containing the compound of the present invention, the selected compound may be mixed with solid, powdered ingredients such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose derivatives, gelatin, or another suitable ingredient, as well as with loosening agents and lubricating agents such as magnesium stearate, calcium stearate, sodium fumarate and polietilenglikolya waxes. This mixture can then be processed into granules or compressed into tablets.

Soft gelatin capsules may be prepared with capsules containing a mixture of active compound or compounds according to the present invention, vegetable oil, grease or other under tamago filler for soft gelatin capsules. Hard gelatin capsules may contain granules of the active compound. Hard gelatin capsules may contain the active compound in combination with solid powdered ingredients such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives or gelatine.

Dosage units for rectal injection can be prepared (1) in the form of suppositories which contain the active substance is mixed with a neutral fat base; (2) in the form of a gelatine rectal capsules which contain the active substance in a mixture with a vegetable oil, paraffin oil or other suitable filler for gelatin rectal capsules; (3) in the form of ready-made micro; or (4) in the form of a dry preparation for the micro that is diluted in a suitable solvent just before the introduction.

Liquid preparations can be prepared in the form of syrups or suspensions, for example solutions or suspensions containing the active ingredient and the remainder, consisting, for example, of sugar or sugar alcohols and a mixture of ethanol, water, glycerol, propylene glycol and polyethylene glycol. If required, such liquid preparations may contain colouring agents, corrigentov, preservatives, saccharin and carboxymethyl cellulose or other zag is a Fort worth. Liquid preparations can also be made in the form of a dry powder, intended for cultivation in a suitable solvent before use.

Solutions for parenteral administration can be prepared in the form of a solution of the compound of the present invention in a pharmaceutically acceptable solvent. These solutions may also contain stabilizing ingredients, preservatives and/or buffer ingredients. Solutions for parenteral administration can be prepared in the form of a dry product intended for reconstitution in a suitable solvent before use.

A typical daily dose of the active substance varies widely and depends on various factors, such as, for example, an individual need of each patient, the route of administration and the disease. In General, oral and parenteral dosages are in the range from 0.1 to 1000 mg per day of the active substance.

Medical and pharmaceutical application

Compounds according to the invention either by themselves are inhibitors of carboxypeptidase U, or, in the case of prodrugs, become them after injection. Thus, it is expected that the compounds according to the invention will be useful in those conditions in which inhibition of carboxypeptidase U is beneficial, for example in the treatment or prevention the ICA thrombosis or hypercoagulability in blood and tissues of mammals, including humans.

It is known that hypercoagulation may lead to thromboembolic disease. Conditions associated with hypercoagulation and thromboembolic diseases that may be mentioned include resistance to protein C and hereditary or acquired deficiencies of anti-thrombin III, protein C, protein S and heparin cofactor II. Other conditions about which it is known that they are associated with hypercoagulation or thromboembolic disease include circulatory and septic shock, circulating antiphospholipid antibodies, homocysteinemia, heparin-induced thrombocytopenia and defects in fibrinolysis. Thus, the compounds according to the invention is shown both for therapeutic and/or prophylactic treatment of these conditions. Compounds according to the invention are also indicated for the treatment of conditions in which occurs an undesirable excess of CPU/CPU.

Specific painful conditions that can be referred to as intended for therapeutic and/or prophylactic treatment include venous thrombosis and pulmonary embolism, arterial thrombosis (e.g. in myocardial infarction, unstable angina, stroke due to thrombosis and peripheral arterial thrombosis) and systemic embolism usually from the atrium during arterial fibrillate the or from the left ventricle after transmural myocardial infarction.

In addition, it is expected that the compounds according to the invention will have application for the prevention of reocclusion or restenosis (i.e. thrombosis) after thrombolyse, percutaneous transluminal angioplasty (MOUTH) and coronary artery bypass operations; the prevention of re-thrombosis after microsurgery surgery and vascular surgery in General.

Additional indications include therapeutic and/or prophylactic treatment of disseminated intravascular coagulation caused by bacteria, multiple trauma, intoxication or any other mechanism of anticoagulant treatment when blood is in contact with foreign surfaces in the body such as vascular grafts, vascular stents, vascular catheters, mechanical and biological prosthetic valves or any other medical device, and anticoagulant treatment when blood is in contact with medical devices outside the body, such as used during cardiovascular surgery apparatus "heart-lung or devices used in hemodialysis.

The connection according to the invention can also be combined and/or co to enter with any antithrombotic agent with a different mechanism of action, such as antithrombotic agents acetylsalicylic sour is a, ticlopidine, clopidogrel, an inhibitor of thromboxane receptor and/or synthetase, receptor antagonists of fibrinogen, prostacyclin mimetics and phosphodiesterase inhibitors and antagonists of ADP-receptor (R2T) (ADP - adenosine-5’-diphosphate), and thrombin inhibitors.

Compounds according to the invention can be combined and/or co to enter with thrombolytics such as tissue activator of plasminogen (natural, recombinant or modified), streptokinase, urokinase, PUK, Antilibanus the plasminogen-streptokinase activator complex (APSAC), plasminogen activators salivary glands of animals and the like, in the treatment of thrombotic diseases, in particular myocardial infarction and stroke.

In vitro experiments

Inhibitory effect of compounds of the present invention was evaluated using the analysis described in Dirk Hendriks, Simon Scharpe and Marc van Sandre, Clinical Chemistry, 31, 1936-1939 (1985); and Wei Wang, Dirk F.Hendriks, Simon S.Scharpe, The Journal of Biological Chemistry, 269, 15937-15944 (1994).

Examples

General experimental methods

Mass spectra were taken on a triple quadrupole mass spectrometer Finnigan Mat TSQ 700, equipped with interface elektrorazpredelenie (FAB-MS (ei bombardment of fast atoms-mass spectrometry)), and mass spectrometer VG Platform II, equipped with interface elektrorazpredelenie (LC-MS (liquid chromatography-mass spectrometry)). 1H NMR and13With NMR measurements were performed on the spectrometer Varian UNITY plus 400, 500 and 600 running on1H frequencies of 400, 500 and 600 MHz, respectively. Chemical shifts are given in ppm (ppm) with the solvent as internal standard. The organic extracts were dried using MgSO4or NaSO4as a dewatering agent. Chromatographic separation was performed using Merck Silica gel 60 (0,063-0,200 mm). HPLC (high performance liquid chromatography) separation was performed on a column of HIGHCROM KR100-10C8.

Example 1

2-Mercaptomethyl-3-piperidine-4-yl-propionic acid

a) 3-piperidine-4-yl-propionic acid

A solution of 3-pyridin-4-yl-acrylic acid (4,20 g of 28.0 mmol) in water (50 ml) and ammonia (water, 25%, 4 ml) was first made at 60 bar (60·105PA) in steel high pressure autoclave in the presence of ruthenium (5% aluminum oxide, 439 mg). When the hydrogen pressure remained constant (3 days), the catalyst was removed from the reaction mixture by filtration. The catalyst was washed with ethanol and water, and the ethanol was removed from the solution on a rotary evaporator, and the aqueous solution was dried by sublimation with 3-piperidine-4-yl-propionic acid (4,30 g, 100%).

b) 4-(2-Carboxy-ethyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

A solution of 3-piperidine-4-yl-propionic acid (4,79 g of 30.5 mmol), di-tert-boutillier the Nata (6,98 g, of 32.0 mmol) and NaHCO3(2,69 g of 32.0 mmol) in a mixture of THF/water (1:1, 50 ml) was stirred at room temperature for 22 hours. Another portion of di-tert-BUTYLCARBAMATE (2.00 g, 9,10 mmol) was added along with a catalytic amount of DMAP, the mixture was stirred for another four days. THF was removed under reduced pressure, and the aqueous phase was extracted with CH2Cl2. The aqueous phase is then acidified to pH 2 1 M HCl, and the acidic phase was extracted with CH2Cl2. The combined organic extracts were washed with brine, dried and concentrated in vacuum to obtain 4-(2-carboxy-ethyl)-piperidine-1-carboxylic acid tert-butyl ester as a white solid (6,36 g, 81%).

b) 4-(3-Benzylmorphine-2-carboxy-propyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

BuLi (1,6 M, 15.3 ml, 24.4 mmol) was added to a solution of Diisopropylamine fees (3.43 ml, 24.4 mmol) in THF (3 ml) at -78°C in argon atmosphere. After several minutes, the solution was left to warm to room temperature over a period of time of 15 minutes. The resulting LDA solution was slowly added to a solution of 4-(2-carboxy-ethyl)-piperidine-1-carboxylic acid tert-butyl ester (of 3.07 g of 11.9 mmol) in THF (7 ml) at -78°C. the resulting solution was stirred at -78°C for 10 minutes. At this time, was added THF (20 ml) to dissolve the anion, which is first hardened. Dianion was cooled to -78°and added promotility ether (2,72 g, 12.5 mmol) in solution in THF (3 ml). The solution was stirred at -78°C for 30 minutes, at 0°C for 30 minutes and then left to warm to room temperature and was stirred overnight. The reaction mixture was acidified using 1 M HCl, diluted tO, and the organic phase is washed with water and dried. The crude product was purified flash chromatography (Meon/l3, 1:9) to obtain 4-(3-benzylmorphine-2-carboxy-propyl)-piperidine-1-carboxylic acid tert-butyl ester as a pale yellow oil (3.12 g, 66%).

g) 4-(2-Carboxy-3-mercapto-propyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

Sodium metal (513 g of 22.5 mmol) was added in portions over 5 minutes to a solution of 4-(3-benzylmorphine-2-carboxy-propyl)-piperidine-1-carboxylic acid tert-butyl ester (0.9 g, to 2.29 mmol) in THF and aqueous ammonia (50 ml) at -69°C in argon atmosphere. After stirring for 15 minutes portions were added ammonium chloride (1.7 g, to 31.5 mmol). The cooling bath was removed, and the ammonia evaporated using a stream of argon. Was added 0.5 M NaOH, and the mixture was washed with heptane. The aqueous phase was acidified using 2 M HCl and was extracted with methylene chloride. The organic phase was washed with brine, dried and concentrated under reduced pressure to obtain 4-(2-carboxy-3-mercapto-propyl)-piperidine-1-carboxylic acid tert-butyl ester (0.7 g, 100%).

d) 2-Mercaptomethyl-3-piperidine-4-yl-propionic acid

To a solution of 4-(2-carboxy-3-mercapto-propyl)-piperidine-1-carboxylic acid tert-butyl ester (0.7 g, to 2.29 mmol) in methylene chloride (8 ml) in an argon atmosphere was added triethylsilane (731 μl, 4,58 mmol), then TFA (4 ml). The reaction mixture was stirred for 60 minutes and then concentrated under reduced pressure. After HPLC purification (10→30% acetonitrile, 0.1% of TFA in water) has been specified in the title compound as TFA salt (447 mg, 61%).

1H NMR (400 MHz, D2O): δ 1.34-1.50 (m, 2H), 1.54-1.76 (m, 3H), 1.90-1.99 (m, 1H), 2.0-2.1 (m, 1H), 2.9-3.05 (m, 5H), 3.38-3.48 (m, 2H).

MS(+) 204(M+1).

Example 2

3-(1-Acetyl-piperidine-4-yl)-2-mercaptomethyl-propionic acid

A solution of the TFA salt of 2-mercaptomethyl-3-piperidine-4-yl-propionic acid (0.1 g, 0.32 mmol) in acetic anhydride (2 ml) was stirred overnight in an argon atmosphere, and then concentrated under reduced pressure. After HPLC purification (10→50% acetonitrile, 0.1% of TFA in water) has been specified in the title compound (63 mg, 80%).

1H NMR (500 MHz, CD3OD): δ 1.0-1.22 (m, 2H), 1.46-1.55 (m, 1H), 1.55-1.66 (m, 2H), 1.67-1.79 (m, 1H), 1.81-1.92 (m, 1H), 2.08 (s, 3H), 2.55-2.73 (m, 4H), 3.03-3.12 (m, 1H), 3.85-3.94 (m, 1H), 4.45-4.53 (m, 1H).

MS(+) 246(M+1).

Example 3

3-Mercapto-5-methyl-2-piperidine-4-ylmethyl-hexanoic acid

a) 4-Hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ether

To a solution of 4-(hydroxymethyl)piperidine (5,00 g, 43,41 mmol in THF/H2O (1:1, 120 ml) was added di-tert-BUTYLCARBAMATE (for 9.47 g, 43,41 mmol). The reaction mixture was stirred for 16 hours at room temperature. The reaction mixture then was poured into H2O (500 ml) and was extracted with ethyl acetate (3×250 ml). The organic layers were combined and washed with water. The organic layer was dried over sodium sulfate, filtered, and then concentrated under reduced pressure to obtain 4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ether (9,01 g, 96%).

b) 4-methyl bromide-piperidine-1-carboxylic acid tert-butyl methyl ether

To a solution of 4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ether (8,75 g, 40,64 mmol) in diethyl ether (200 ml) at 0°C in an atmosphere of nitrogen was added triphenylphosphine (one-21.32 g, to 81.28 mmol) and tetrabromide carbon (26,96 g to 81.28 mmol). The mixture was left to warm to room temperature and stirred under nitrogen atmosphere for 48 hours. The reaction mixture was filtered through a layer of Celite, and the organic filtrate is washed with 5% PA2S2O3, water, brine and dried. The mixture was filtered and concentrated under reduced pressure. The crude product was purified column chromatography (ethyl acetate/hexane, 1:9) to obtain 4-methyl bromide-piperidine-1-carboxylic acid tert-butyl EF is RA (compared to 8.26 g, 73%).

C) 4-[2-tert-Butoxycarbonyl-2-(diethoxy-phosphoryl)-ethyl]-piperidine-1-carboxylic acid tert-butyl methyl ether

tert-Butyldichlorosilane (75,0 g, 297,32 mmol) was added dropwise to a suspension of sodium hydride (8,03 g, 334,58 mmol) in DMF (450 ml) at 0°C in nitrogen atmosphere. The mixture was stirred at 0°C for 0.5 hours and at room temperature for 0.5 hours. 4-methyl bromide-piperidine-1-carboxylic acid tert-butyl ether (20,68 g, 74,34 mmol) in DMF (50 ml) was added dropwise to the reaction mixture, and the reaction mixture was heated to 60°and was stirred for 16 hours. The reaction mixture was cooled to room temperature, poured into H2O and was extracted with ethyl acetate. The organic layers were combined and washed with water. The organic layer was dried, filtered and concentrated under reduced pressure. The crude product was purified column chromatography (ethyl acetate/hexane, 3:7) to give 4-[2-tert-butoxycarbonyl-2-(diethoxy-phosphoryl)-ethyl]-piperidine-1-carboxylic acid tert-butyl ether and unreacted tert-butyldiethanolamine. According to HPLC the product had a purity of 47%. This product is then purified by vacuum distillation to a purity of 77%. This mixture was used in the next reaction.

g) 4-(2-tert-Butoxycarbonyl-5-methyl-Gex-2-enyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

4-[2-is pet-Butoxycarbonyl-2-(diethoxy-phosphoryl)-ethyl]-piperidine-1-carboxylic acid tert-butyl ester (8.1 g) in 20 ml of DME (1,2-dimethoxyethane) was added dropwise to a suspension of sodium hydride (1.04 g, 43,13 mmol) in DME (20 ml) at 0°C in nitrogen atmosphere. The mixture was stirred for 0.75 hour, and to the mixture was added dropwise isovaleraldehyde (7,76 g, 90,1 mmol). The mixture was left to warm to room temperature and was stirred for 48 hours. The mixture was diluted with ether and washed with water. The organic layer was dried, filtered and concentrated under reduced pressure to obtain 9.4 g of a yellow oil. The crude product was purified column chromatography (ethyl acetate/hexane, 1:50) to give 4-(2-tert-butoxycarbonyl-5-methyl-Gex-2-enyl)-piperidine-1-carboxylic acid tert-butyl ether (1,53 g, 24%) for the two reactions.

d) 4-[2-tert-Butoxycarbonyl-3-(4-methoxy-benzylmethyl)-5-methyl-hexyl]-piperidine-1-carboxylic acid tert-butyl methyl ether

A solution of 4-(2-tert-butoxycarbonyl-5-methyl-Gex-2-enyl)-piperidine-1-carboxylic acid tert-butyl ester (2.0 g, of 5.24 mmol) in DMF (5 ml) was added to a mixture of potassium carbonate (0.54 g, 3.93 mmol) and 4-methoxy-α-colortool (1,17 g, 10,48 mmol) in DMF (50 ml) under nitrogen atmosphere. The mixture was boiled under reflux for 5 hours and left to cool to room temperature. The reaction mixture then was poured into H2O and was extracted with ethyl acetate. The organic layers were combined and washed with water. The organic layer was dried, filtered and concentrated under reduced pressure to receive the receiving 3,76 g of the crude substance. The crude product was purified column chromatography (ethyl acetate/hexane, 1:10) to give 4-[2-tert-butoxycarbonyl-3-(4-methoxy-benzylmethyl)-5-methyl-hexyl]-piperidine-1-carboxylic acid tert-butyl ester (1.77 g, 63%).

e) 3-Mercapto-5-methyl-2-piperidine-4-ylmethyl-hexanoic acid

A mixture of N2O (2,6 ml) and TFA (26 ml) was frozen, and then left to warm to room temperature in nitrogen atmosphere. Was added 4-[2-tert-butoxycarbonyl-3-(4-methoxy-benzylmethyl)-5-methyl-hexyl]-piperidine-1-carboxylic acid tert-butyl ether (2,62 g, 4,89 mmol), and the mixture is boiled under reflux for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The crude product was purified column chromatography with reversed phase (MeOH/H2O 3:2) to obtain specified in the title compounds as TFA salt (0.40 g, 22%).

1H NMR (300 MHz) (CD3OD): δ 0.88 (d), 0.94 (d), 1.43 (m), 1.70 (br), 1.94 (m), 2.48 (m), 2.90 (m), 2.99 (m), 3.34 (m).

MS (+) OF 260.2 (M-TFA).

Example 4

3-Mercapto-4-phenyl-2-piperidine-4-ylmethyl-butyric acid

a) 4-(2-tert-Butoxycarbonyl-4-phenyl-but-2-enyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

A solution of 4-[2-tert-butoxycarbonyl-2-(diethoxy-phosphoryl)-ethyl]-piperidine-1-carboxylic acid tert-butyl ester (3.0 g) in DME (8 ml) was added to cap the holes to a suspension of sodium hydride (0.26 g, 10,51 mmol) in DME (8 ml) at 0°C in nitrogen atmosphere. The mixture was stirred for 0.75 in hours. Phenylacetaldehyde (5,26 g, 43,81 mmol) was added dropwise to the mixture at 0°C. the Mixture was left to warm to room temperature and was stirred for 16 hours. The mixture was diluted with ether and washed with water. The organic layer was dried, filtered and concentrated under reduced pressure to obtain 8.6 g of a yellow oil. The crude product was purified column chromatography (ethyl acetate/hexane, 1:50→1:10) to give 4-(2-tert-butoxycarbonyl-4-phenyl-but-2-enyl)-piperidine-1-carboxylic acid tert-butyl ether (1,32 g, yield 62% for the two reactions).

b) 4-[2-tert-Butoxycarbonyl-3-(4-methoxy-benzylmethyl)-4-phenyl-butyl]-piperidine-1-carboxylic acid tert-butyl methyl ether

4-(2-tert-Butoxycarbonyl-4-phenyl-but-2-enyl)-piperidine-1-carboxylic acid tert-butyl ester (0.8 g, of 1.93 mmol) in DMF (10 ml) was added to a suspension of potassium carbonate (0.20 g, 1.44 mmol) and 4-methoxy-α-colortool (0.54 ml of 3.85 mmol) in DMF (10 ml) under nitrogen atmosphere. The mixture was heated to 75°C for 24 hours and left to cool to room temperature. The reaction mixture then was poured into H2O and was extracted with ethyl acetate. The organic layers were combined and washed with water. The organic layer was dried, filtered and concentrated under reduced pressure to half the rising 1.8 g of the crude substance. The crude product was purified column chromatography (ethyl acetate/hexane, 1:10) to give 4-[2-tert-butoxycarbonyl-3-(4-methoxy-benzylmethyl)-4-phenyl-butyl]-piperidine-1-carboxylic acid tert-butyl ester (0.55 g, 50%).

C) 3-Mercapto-4-phenyl-2-piperidine-4-ylmethyl-butyric acid

A mixture of H2O (0,65 ml) and TFA (6.5 ml) was frozen, and then left to warm to room temperature in nitrogen atmosphere. Was added 4-[2-tert-butoxycarbonyl-3-(4-methoxy-benzylmethyl)-4-phenyl-butyl]-piperidine-1-carboxylic acid tert-butyl ester (0.65 g, to 1.14 mmol)and the mixture is boiled under reflux for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The crude product was purified column chromatography with reversed phase (MeOH/H2O, 1:1) to obtain specified in the title compounds as TFA salt (0.27 g, 58%).

1H NMR (300 MHz) (SO): δ 1.22 (m), 1.49 (m), 1.67 (br), 2.21 (d), 2.98 (m), 7.25 (m, 5H), 8.27 (br), 8.57 (br), 12.59 (br).

MS (+) 294,3 (M-TFA).

Example 5

2-(2-Amino-pyridine-4-ylmethyl)-3-mercapto-propionic acid

a) N-(4-Methyl-pyridine-2-yl)-ndimethylacetamide

2-Amino-4-methylpyridin (a 99.0 g of 91.5 mmol) in acetic anhydride (250 ml) was heated to 70°C for two hours. The mixture was cooled to room temperature and was added diethyl ether (100 ml). The product has led in the de white needle-shaped crystals. After filtering and drying in vacuum is obtained N-(4-methyl-pyridine-2-yl)-ndimethylacetamide (130 g, 95%).

b) 2-Acetylamino-isonicotinoyl acid

A mixture of N-(4-methyl-pyridine-2-yl)-ndimethylacetamide (40,0 g, 0.26 mmol) and water (400 ml) was heated at 90°until the solution became homogeneous. Cautiously in small portions was added KMPO4(100 g of 0.62 mmol) under intensive mechanical stirring for 2 hours. The reaction mixture was stirred at 90-95°C for another 3 hours and then filtered through Celite, while it was still hot. The filtrate was concentrated to approximately 100 ml, and was added concentrated HCl to bring the pH to approximately 4. The reaction flask was cooled in an ice bath, and the white solid was filtered. The crystals were washed with cold water and chloroform and dried in vacuum to obtain 2-acetylamino-isonicotinic acid (12.0 g, 25%).

a) 2-Amino-isonicotinic acid ethyl ester

2-Acetylamino-isonicotinoyl acid (10.8 g, 60,0 mmol) suspended in ethanol (150 ml) was added BF3OEt2(22 ml, 138 mmol). The mixture was boiled under reflux during the night, and after cooling to room temperature was added 10% NaHCO3(250 ml). The product was extracted with chloroform, and the combined organic extracts were washed with water and dried. After filtration and concentration obtained is 2-amino-isonicotinic acid ethyl ester (7,46 g, 79%) as pale yellow crystals.

g) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-isonicotinic acid ethyl ester

To a solution of 2-amino-isonicotinic acid ethyl ester (of 5.00 g, 30 mmol) in tert-BuOH (45 ml) and acetone (15 ml) was added DMAP (50 mg, 0.41 mmol) and di-tert-BUTYLCARBAMATE (16.4 g, 75,0 mmol). The reaction mixture was stirred at room temperature over night and was added hexane (60 ml). The reaction mixture was cooled in the refrigerator for 3 hours and filtered to obtain 2-[N,N-bis(tert-butoxycarbonyl)amino]-isonicotinic acid ethyl ester (8,71 g, 79%).

d) (4-Hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

A solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-isonicotinic acid ethyl ester (35,0 g, 95.5 mmol) in THF (350 ml) was treated with LiAlH4(7,25 g, 191 mmol) and boiled under reflux for 1 hour in nitrogen atmosphere. The reaction mixture was carefully poured onto crushed ice and the product was extracted several times l3and l3:Meon (9:1). The combined organic extracts were dried, filtered and concentrated under reduced pressure to obtain (4-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ester (18.5 g, 86%) as a pale yellow solid.

e) (4-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

(4-G is proximity-pyridine-2-yl)-carbamino acid tert-butyl ester (8.00 g, of 35.6 mmol) was dissolved in CH2Cl2(150 ml) and treated h3(11.2 g, 42.8 mmol) under nitrogen atmosphere. The reaction flask was cooled in an ice bath and small portions were added GVS4(14.2 g, 42.8 mmol). The ice bath was removed after 30 minutes and the reaction mixture was stirred over night at room temperature. The reaction mixture was concentrated under reduced pressure and added to acetonitrile (50 ml). The reaction flask was placed in a refrigerator for 3 hours, and the precipitate was filtered and washed with cold acetonitrile. White solid was dried in vacuum to obtain (4-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ether (scored 8.38 g, 82%).

W) 2-(2-tert-Butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid diethyl ester.

To a solution of NaH (80%, 0.17 g, 4.00 mmol) in THF (5 ml) at 0°C in an atmosphere of argon was added diethylmalonate (0.64 g, 4.00 mmol). After stirring the mixture for 15 minutes the mixture was added to the heated to the temperature of reflux distilled mixture of (4-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ether (1,00 g of 3.48 mmol) in THF (10 ml)and the mixture is boiled under reflux for 2 hours. The mixture was concentrated under reduced pressure, and the residue was distributed between water and chloroform. The organic layer was washed with water and brine and dried. After filtration and evaporation of the races is vorites the crude product was purified flash chromatography (MeOH/CH 2Cl2, 1:100) to give 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid diethyl ester (0,80 g, 55%).

C) 2-(2-tert-Butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid monotropy ether

A solution of KOH (0,19 g of 3.43 mmol) in ethanol (5 ml) was added to a solution of 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid diethyl ester (1.20 g, with 3.27 mmol) in ethanol (5 ml) and methylene chloride (5 ml) at 0°C. the Mixture was stirred for 18 hours at room temperature. The mixture was concentrated under reduced pressure and to the residue was added water. The aqueous layer was washed with diethyl ether, acidified to pH 4 1 M HCl and was extracted with methylene chloride. The organic layer was washed with water, brine and dried. After filtration and evaporation in vacuo the crude product was purified flash chromatography (CH3OH/CH2Cl2, 1:20) to give 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid of monoethylene ether (0,90 g, 81%).

and) 2-(2-tert-Butoxycarbonylamino-pyridine-4-ylmethyl)-acrylic acid ethyl ester

The solution diethylamine (0.26 g, to 2.67 mmol) in methylene chloride (4 ml) was added to a mixture of 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-malonic acid of monoethylene ether (0,90 g of 2.66 mmol) and 37% aqueous formaldehyde solution (0.24 g, 3.00 mmol) at 0°C. the Mixture was stirred is for 3 hours at room temperature, poured into ice water and was extracted with methylene chloride. The organic layer is washed with 5% Panso3and dried. The crude product was purified flash chromatography (1% methanol in CH2Cl2) to obtain 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-acrylic acid ethyl ester (0,58 g, 71%).

K) 2-Acetylsalicylate-3-(2-tert-butoxycarbonylamino-pyridin-4-yl)-propionic acid ethyl ester

A solution of 2-(2-tert-butoxycarbonylamino-pyridine-4-ylmethyl)-acrylic acid ethyl ester (0,48 g, 1.57 mmol) and triethylamine (0.17 g, of 1.64 mmol) was added to teoksessa acid (3 ml) at 0°C in nitrogen atmosphere. The mixture was stirred at room temperature for 4 hours. The mixture was poured into ice water and was extracted with CH2Cl2. The organic phase was washed with saturated NaHCO3(water) and dried. The crude product was purified flash chromatography (2,5%Meon in CH2Cl2) to obtain 2-acetylsalicylate-3-(2-tert-butoxycarbonylamino-pyridin-4-yl)-propionic acid ethyl ester (0,60 g, 100%).

l) 2-Acetylsalicylate-3-(2-amino-pyridine-4-yl)-propionic acid ethyl ester

TFA (0.5 ml) was added to a solution of 2-acetylsalicylate-3-(2 - tert-butoxycarbonylamino-pyridin-4-yl)-propionic acid ethyl ester (50 mg, 0.13 mmol) in methylene chloride in an argon atmosphere. The solution paramesh the Wali for 60 minutes and then concentrated under reduced pressure to obtain crude 2-acetylsalicylate-3-(2-amino-pyridine-4-yl)-propionic acid ethyl ester (52 mg, 100%).

1H NMR (500 MHz, CD3OD): δ 1.15 (t, 3H), 2.32 (s, 3H), 2.73-2.83 (m, 2H), 2.86-2.93 (m, 1H), 3.01-3.07 (dd, 1H), 3.12-3.18 (dd, 1H), 4.03-4.12 (m, 2H), 6.39 (s, 1H), 6.43 (d,1H), 7.77 (d,1H).

m) 2-(2-Amino-pyridine-4-ylmethyl)-3-mercapto-propionic acid

2-Acetylsalicylate-3-(2-amino-pyridine-4-yl)-propionic acid ethyl ester (52 mg, 0.13 mmol) was dissolved in concentrated HCl (2 ml) in an argon atmosphere. The solution was heated to the temperature of reflux distilled for 1 hour. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (32 mg, 100%).

1H NMR (500 MHz, CD3OD): δ 2.70 (bs, 2H), 2.85-3.0 (m, 3H), 6.76 (bs, 1H), 6.81 (bs, 1H), 7.67 (bs, 1H).

MS(+) 213(M+1).

Example 6

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-propionic acid

a) 6-Aminonicotinic acid ethyl ester

2-Amino-5-pyridylcarbonyl acid (25,0 g, 181 mmol) suspended in ethanol (190 ml) was added SOCl2(15 ml, 206 mmol). The mixture was boiled under reflux for 10 hours and was added SOCl2(16 ml). After 3 hours boiling under reflux (and adding another SOCl2(10 ml) every day), the reaction mixture was cooled to room temperature and was added diethyl ether. After 24 hours at -20°the mixture was filtered. The crude salt was dissolved in methanol (214 ml) was added NaOH solution (40,0 g, 23.5 mmol) in methanol is (90 ml). The reaction mixture was stirred for 1 hour and was added THF (270 ml). The reaction mixture was filtered through a gasket made of silicon dioxide (THF/MeOH) and concentrated under reduced pressure to obtain 6-amino-nicotinic acid ethyl ester (36.2 g, 97%).

b) 6-tert-Butoxycarbonylamino-nicotinic acid ethyl ester

To a solution of 6-amino-nicotinic acid ethyl ester (36,0 g, 217 mmol) in tert-BuOH (308 ml) and acetone (103 ml) was added DMAP (0,53 g, 4,34 mmol) and di-tert-BUTYLCARBAMATE (72,0 g, 330 mmol). The reaction mixture was stirred at room temperature for 10 hours, then was added di-tert-BUTYLCARBAMATE (2,60 g). After 10 hours of stirring at room temperature was added hexane (470 ml). The reaction mixture was cooled to -20°C for 2 hours and filtered. The filtrate was washed with hexane/dichloromethane (3:1) and dried in vacuo to obtain 6-tert-butoxycarbonylamino-nicotinic acid ethyl ester (40,5 g, 70%).

in) (5-Hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

To a stirred solution of 6-tert-butoxycarbonylamino-nicotinic acid ethyl ester (3.50 g, of 13.1 mmol) in THF (20 ml) under nitrogen atmosphere was added LiAlH4(0,91 g of 24.0 mmol) in THF (20 ml) over a time period of 2 hours. The reaction mixture was stirred for 6 hours, then was added (cautiously) NH4Cl (the seal) as long until the solution becomes neutral. The mixture was filtered and concentrated under reduced pressure to obtain (5-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ester (2.00 g, 68%).

g) (5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

Triphenylphosphine (to 8.70 g, up 33.1 mmol) and tetrabromide carbon (17.0 g, a 51.2 mmol) was added to a suspension of (5-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ether (7,00 g, and 31.2 mmol) in CH2Cl2(200 ml) at room temperature. Stirring was continued for 5 hours, then the solvent evaporated. Added acetonitrile (200 ml)and the mixture was cooled to -20°C for 2 hours. The mixture is then filtered, and the crystalline residue was washed with cold acetonitrile (2×10 ml) to give (5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ether (5,96 g, 67%).

d) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid diethyl ester

To a suspension of NaH (0,49 g, 16.3 mmol, 80%) in THF (15 ml) at 0°With added diethylmalonate (2,61 g, 16.3 mmol). The mixture was stirred for 15 minutes and then added dropwise to the heated to the temperature of reflux distilled mixture of (5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ether (3,90 g of 13.6 mmol) in THF (25 ml)and the resulting solution was boiled under reflux for 15 minutes. PEFC is the evaporation of the solvent the crude product was purified flash chromatography (methanol/CH 2CL2that 1:100 to 2.5:100) to give 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid diethyl ester (2,18 g, 44%).

e) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid monotropy ether

A solution of KOH (0,37 g is 6.54 mmol) in ethanol (5 ml) was added to a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid diethyl ester (2,18 g, 5,95 mmol) in ethanol (25 ml) and methylene chloride (10 ml) at 0°C. the Mixture was stirred for 18 hours at room temperature. This mixture was concentrated under reduced pressure, and the residue was dissolved in water. The aqueous layer was washed with ether, acidified to pH 4 1 M HCl and was extracted with methylene chloride. The organic layer was washed with water, brine and dried. After filtration and concentration under reduced pressure the crude product was purified flash chromatography (methanol/CH2CL2, 1:20) to give 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid of monoethylene ether (1,00 g, 50%).

W) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-acrylic acid ethyl ester

Diethylamine (0,29 g, 3.00 mmol), water (2 ml) and methylene chloride (2 ml) was added to a mixture of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-malonic acid of monoethylene ether (1,00 g, 2,96 mmol) and 37% aqueous formaldehyde (0.25 g, of 3.05 mmol) at 0°C. Masi was stirred for 16 hours at room temperature, and then poured into ice water and was extracted with methylene chloride. The organic layer is washed with 5% Panso3and dried. After filtration and concentration under reduced pressure was obtained 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0.75 g, 83%).

C) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propionic acid ethyl ester

2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0,49 g to 1.60 mmol) and triethylamine (0.17 g, of 1.64 mmol) was added to teoksessa acid (3 ml) at 0°C. the Mixture was stirred at room temperature for 6 hours. This mixture is poured into ice water and was extracted with CH2CL2. The organic phase was washed with saturated NaHCO3and dried. The crude product was purified flash chromatography (Meon/CH2CL2, 2,5:100) to give 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propionic acid ethyl ester (0.36 g, 61%).

and) 2-Acetylsalicylate-3-(6-amino-pyridin-3-yl)-propionic acid ethyl ester

TFA (0.5 ml) was added to a solution of 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propionic acid ethyl ester (100 mg, 0.26 mmol) in methylene chloride (2 ml) in an argon atmosphere. The solution was stirred for 60 minutes and then concentrated in on igenom pressure to obtain crude 2-acetylsalicylate-3-(6-amino-pyridin-3-yl)-propionic acid ethyl ester (104 mg, 100%).

1H NMR (500 MHz, CD3OD): δ 1.21 (t, 3H), 2.33 (s, 3H), 2.78-2.97 (m, 3H), 3.05-3.13 (m, 1H), 3.14-3.21 (m, 1H), 4.08-4.15 (m, 2H), 6.99 (d, 1H), 7.69 (s, 1H), 7.85 (d, 1H).

K) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-propionic acid

2-Acetylsalicylate-3-(6-amino-pyridin-3-yl)-propionic acid ethyl ester (38 mg, 0,096 mmol) was dissolved in concentrated HCl (2.0 ml) in an argon atmosphere. The solution was stirred at room temperature for 1 hour and then heated to the temperature of reflux distilled for 1 hour. After concentration under reduced pressure has been specified in the header of the connection (of 25.7 mg, 100%) in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, CD3OD): δ 2.74-2.78 (m, 2H), 2.84-2.94 (m, 3H), 7.02 (d, 1H), 7.72 (s, 1H), 7.89 (d, 1H).

MS(+) 213(M+1).

Example 7

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-2-methyl-propionic acid

a) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid tert-butyl ester ethyl ester

A solution of tert-butylethylenediamine (457 g of 2.26 mmol) in DMF (4 ml) was added dropwise to a suspension of NaH (90 mg, and 2.26 mmol, 60% in oil) in DMF (4 ml). The reaction mixture was stirred for 20 minutes. Solution was added (5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ester (500 mg, of 1.74 mmol) in DMF (2.5 ml), and this reaction mixture was stirred for 70 minutes. Added tO, and the mixture was washed with water and R is SOLOM, was dried and concentrated under reduced pressure. After chromatography (heptane/tO, 3:1 1:3) was obtained 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid tert-butyl ester ethyl ester (437 mg, yield 61%).

b) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid mono-tert-butyl ether

1 M NaOH (2 ml) was added to a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid tert-butyl ester ethyl ester (0,42 g of 1.03 mmol) in THF/EtOH (4 ml, 1:1). The reaction mixture was stirred at 50°C for 16 hours. Added CH2Cl2and this mixture was washed with 0.5 M Hcl and brine and dried. After concentration under reduced pressure was obtained 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid mono-tert-butyl ester (348 mg, 89%).

C) 3-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-hydroxymethyl-2-methyl-propionic acid tert-butyl methyl ether

Methylchloroform (75 μl, of 0.92 mmol) was added dropwise to a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-methyl-malonic acid mono-tert-butyl ester (348 mg, 0,915 mmol) and Et3N (123 μl, of 0.92 mmol) in THF (6 ml). The reaction mixture was stirred for 20 minutes, filtered and added dropwise to a suspension of NaBH4(39 mg, 1.04 mmol) in THF (6 ml) at 0°C. the Reaction mixture is eremetical for 16 hours at room temperature, was added 0.2 M HCl, then tO. The organic phase was washed with brine and dried. After concentration under reduced pressure and subsequent chromatography (toluene/tO, 3:1 1:3) was obtained 3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-hydroxymethyl-propionic acid tert-butyl ester (190 mg, 57%).

g) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propionic acid tert-butyl methyl ether

Diethylazodicarboxylate (160 μl, 1.01 mmol) was added dropwise to a solution of 3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-hydroxymethyl-propionic acid tert-butyl ester (180 mg, 0.49 mmol) and triphenylphosphine (266 mg, 1.01 mmol) in THF (6 ml), and this reaction mixture was stirred for 5 minutes. Added toluxury acid (96 μl, of 1.34 mmol)and the reaction mixture was stirred for 16 hours. After concentration under reduced pressure and subsequent chromatography (toluene/tO, 10:1 to 1:1) received 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propionic acid tert-butyl ester (137 mg, 65%).

d) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-2-methyl-propionic acid

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propionic acid tert-butyl ester (4 mg, 9.4 μmol) was dissolved in concentrated HCl in argon atmosphere. The solution was heated with bratim refrigerator for 1 hour. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (2.5 mg, 100%).

1H NMR (500 MHz, CD3OD): δ 1.20 (s, 3H), 2.62 (d, 1H), 2.76-2.83 (m, 2H), 2.95 (d, 1H), 6.94 (d, 1H), 7.64 (d, 1H), 7.80 (dd, 1H).

MS(+) 227 (M+1).

Example 8

2-(6-Amino-pyridine-3-ylmethyl)-2-mercaptomethyl-butyric acid

a) [5-(5-Ethyl-2,2-dimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine-2-yl]-carbamino acid tert-butyl methyl ether

(5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ester (1.0 g, of 3.48 mmol) was added to a solution of 2,2-dimethyl-5-ethyl-1,3-dioxane-4,6-dione (600 mg, of 3.48 mmol) and triethylamine (0.51 ml, 3,66 mmol) in dimethyl sulfoxide (40 ml) under nitrogen atmosphere. The reaction mixture was stirred overnight and was added water (100 ml). The precipitate was filtered and received [5-(5-ethyl-2,2-dimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine-2-yl]-carbamino acid tert-butyl ether (1,15 g, 87%).

b) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-2-ethyl-malonic acid monotropy ether

A solution of metallic sodium (140 mg, between 6.08 mmol) in ethanol (20 ml) was added to a solution of [5-(5-ethyl-2,2-dimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine-2-yl]-carbamino acid tert-butyl ether (1,15 g, 3.04 from mmol) in ethanol (10 ml). The reaction mixture was stirred for 90 minutes and then was added methylene chloride. The mixture was washed with 0.5 M HCl,dried and concentrated under reduced pressure to obtain 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-ethyl-malonic acid of monoethylene ether (1,05 g, 95%).

a) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-2-hydroxy-methyl-butyric acid ethyl ester

Methylchloroform (150 μl, of 1.95 mmol) was added dropwise to a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-ethylmalonic acid of monoethylene ester (700 mg, at 1.91 mmol) and Et3N (275 μl, 1.97 mmol) in THF (15 ml) at -20°C in nitrogen atmosphere. The reaction mixture was stirred for 50 minutes, filtered and added dropwise to a suspension of NaBH4(80 mg, 2.1 mmol) in THF (15 ml) at -20°C. the Reaction mixture was stirred for 16 hours at room temperature. Was added 0.2 M HCl, and then methylene chloride. The organic phase was washed with brine and dried. After concentration under reduced pressure followed by chromatography (toluene/tO, 3:1 1:3) was obtained 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-hydroxymethyl-butyric acid ethyl ester (300 mg, 45%).

g) 2-Acetylsalicylate-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester

Diisopropylcarbodiimide (296 μl, 1.53 mmol) was added dropwise to a solution of triphenylphosphine (402 mg, 1.53 mmol) in THF (4 ml) at 0°in argon atmosphere, and this reaction mixture was stirred for 30 minutes. The solution thioglucose acid (109 μl, 1.53 mmol) and 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-2-hydroxymethyl-butyric acid what you're ethyl ester (0.27 g, 0.77 mmol) in THF (2 ml) was added dropwise over 10 minutes. The reaction mixture was stirred for 60 minutes at 0°and then for 16 hours at room temperature. After concentration under reduced pressure followed by chromatography (heptane/tO, 10:1 to 1:1) received 2-acetylsalicylate-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester (193 mg, 61%).

d) 2-(6-Amino-pyridine-3-ylmethyl)-2-mercaptomethyl-butyric acid

2-Acetylsalicylate-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester (12.3 mg, 30 µmol) was dissolved in concentrated HCl (2 ml) in an argon atmosphere. The solution was heated under reflux for 24 hours. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (8,3 mg, 100%).

1H NMR (500 MHz, CD3OD): δ 0.91 (t, 3H), 1.71 (m, 2H), 2.68 (m, 2H), 2.92 (m, 2H), 6.96 (d, 1H), 7.65 (bs, 1H), 7.82 (dd, 1H).

MS(-) 239 (M-1).

Example 9

3-(6-Amino-5-methyl-pyridin-3-yl)-2-mercaptomethyl-2-methyl-propionic acid

a) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-3-methyl-5-(2,2,5-trimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine

2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-methyl bromide-3-methyl-pyridine (1.6 g, 4.0 mmol) was added to a solution of 2,2,5-trimethyl-1,3-dioxane-4,6-dione (630 mg, 4.0 mmol) and triethylamine (of 0.58 ml, 4.2 mm is l) in dimethyl sulfoxide (40 ml). The reaction mixture was stirred overnight and was added water (100 ml). The mixture was extracted with tO, the combined organic extracts were washed with water and brine and dried. After concentration under reduced pressure, the obtained crude 2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-5-(2,2,5-trimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine (of 2.06 g).

b) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino-5-methyl-pyridin-3-yl-methyl)-2-methyl-malonic acid monotropy ether

A solution of metallic sodium (184 mg, 8.0 mmol) in ethanol (20 ml) was added to a solution of the crude 2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-5-(2,2,5-trimethyl-4,6-dioxo-[1.3]dioxane-5-ylmethyl)-pyridine (of 2.06 g, ~4.0 mmol) in ethanol (20 ml) in an argon atmosphere. The reaction mixture was stirred for 60 minutes and then was added methylene chloride. The mixture was washed with 0.5 M HCl and brine, dried and concentrated under reduced pressure to obtain crude 2-(6-[N,N-bis(tert-butoxycarbonyl)-amino]-5-methyl-pyridine-3-ylmethyl)-2-methyl-malonic acid of monoethylene ether (1.9 grams).

C) 3-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-2-hydroxymethyl-2-methyl-propionic acid ethyl ester

Methylchloroform (338 μl, 4.4 mmol) was added dropwise to a solution of crude 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-2-methyl-malonic acid mo is outillage ester (1.9 g) and Et 3N (641 μl, 4.6 mmol) in THF (30 ml) at -20°C. the Reaction mixture was stirred for 50 minutes, filtered and added dropwise to a suspension of NaBH4(182 mg, 4.8 mmol) in THF (30 ml) at -20°C. the Reaction mixture was stirred for 16 hours at room temperature, was added 0.5 M HCl, and then methylene chloride. The organic phase was washed with brine and dried. After concentration under reduced pressure followed by chromatography (toluene/tO, 10:1 to 1:3) was obtained 3-(6-[N,N-bis(tert-butoxycarbonyl)-amino]-5-methyl-pyridin-3-yl)-2-hydroxymethyl-2-methyl-propionic acid ethyl ester (885 mg, 49%).

g) 2-Acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)-amino]-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester

Diisopropylcarbodiimide (755 μl, 3,91 mmol) was added dropwise to a solution of triphenylphosphine (1,026 g, 3,91 mmol) in THF (10 ml) at 0°and this reaction mixture was stirred for 30 minutes. The solution thioglucose acid (279 μl, 3,91 mmol) and 3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-2-hydroxymethyl-2-methyl-propionic acid ethyl ester (885 mg, a 1.96 mmol) in THF (5 ml) was added dropwise over 10 minutes. The reaction mixture was stirred for 60 minutes at 0°and then for 16 hours at room temperature. After concentration under reduced pressure followed chromium is ografia (heptane/tO, 10:1 1:3) received a 2-acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester with impurities (1,46 g).

d) 2-Acetylsalicylate-3-(6-amino-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester

The crude 2-acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester (1,46 g) was dissolved in TFA (5 ml) and was stirred for 60 minutes. After concentration under reduced pressure followed by chromatography (toluene/EtOAc, 1:1 1:10 0:1) got 2-acetylsalicylate-3-(6-amino-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester with a small amount of impurities (696 mg, 84%).

e) 3-(6-Amino-5-methyl-pyridin-3-yl)-2-mercaptomethyl-2-methyl-propionic acid

2-Acetylsalicylate-3-(6-amino-5-methyl-pyridin-3-yl)-2-methyl-propionic acid ethyl ester (17 mg, 40 μmol) was dissolved in concentrated HCl (2 ml) in an argon atmosphere. The solution was heated under reflux for 150 minutes. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (to 10.7 mg, 96%).

1H NMR (500 MHz, CD3OD): δ 1.20 (s, 3H), 2.23 (s, 3H), 2.61 (d, 1H), 2.79 (2d, 2H), 2.94 (d, 1H), 7.55 (m, 1H), 7.69 (m, 1H).

MS(+) 241 (M+1).

Example 10

3-Mercapto-2-[(piperidine-4-carbonyl)-AMI is about]-propionic acid

CH2Cl2(70 ml) was added to 4-methoxymethylethoxy resin (7 g, the load L=of 0.91 mmol/g, 6,37 mmol) in an argon atmosphere, the resin was allowed to swell for 10 minutes and was added 2-amino-3-mercapto-propionic acid ethyl ester HCl salt (5.9 g, 32 mmol). Then was added TFA (70 ml) in small portions over 10 minutes. The suspension was shaken at room temperature for 1 hour and concentrated under reduced pressure. When it was almost dry, was added toluene (150 ml)and the mixture was again concentrated under reduced pressure. This procedure was repeated twice. Now yellow resin washed with DMF (3×60 ml), CH2CL2(2×60 ml), TEA: CH2CL2(1:1, 2×60 ml), CH2Cl2(2×60 ml), Meon (2×60 ml) and dried under vacuum overnight.

To calculate the load of 2-amino-3-mercapto-propionic acid ethyl ester in the resin, 50 mg of the product was treated with 10% TFA in CH2CL2within 1 minute, and this procedure was repeated 4 times. The mixture was concentrated under reduced pressure to obtain 2-amino-3-mercapto-propionic acid ethyl ester (9.8 mg), which indicates that the load is approximately 0.6 mmol/g

A solution of piperidine-1,4-dicarboxylic acid mono-tert-butyl ester (28 mg, 0.12 mmol) in DMF (1 ml) was added to the resin (100 mg, L=0.6 mmol/g, 0.06 mmol) plastic syringe, then add the Lyali Rover (62 mg, 0.12 mmol) in DMF (0.5 ml) and DIPEA (41 μl, 0.24 mmol). The reaction mixture was left at room temperature for 2 hours with occasional stirring, and the procedure was repeated again. Then the resin washed with DMF (2×2 ml), CH2Cl2(2×2 ml), Meon (2×2 ml), CH2Cl2(2×2 ml) and THF (2×2 ml). THF (800 μl) was added by syringe, and the resin was allowed to swell for 10 minutes. Then added water (250 μl) and 10 M NaOH (50 μl). The reaction mixture was left to stand at room temperature with periodic mixing. The resin then was washed with a mixture TNR:water (1:1, 2×2 ml), THF (2×2 ml), CH2Cl2(2×2 ml), Meon (2×2 ml) and CH2Cl2(2×2 ml).

10% TFA in CH2CL2(1 ml) was added by syringe, and after 5 minutes the solution was collected in a tared beaker. This procedure was repeated once more and the combined organic phases were concentrated under reduced pressure to obtain specified in the title compounds as TFA salt (15.3 mg, 74%).

1H NMR (500 MHz, CD3OD): δ 1.85-2.10 (m, 4H), 2.65-2.72 (m, 1H), 2.85-2.92 (m, 1H), 2.95-3.08 (m, 3H), 3.40-3.47 (m, 2H), 4.55-4.60 (m, 1H).

Example 11

2-[(Azetidin-2-carbonyl)-amino]-3-mercapto-propionic acid

Specified in the title compound was obtained from azetidine-1,2-dicarboxylic acid 1-tert-butyl ether by the method described in Example 14. Output: 13.8V mg (72%).

1H NMR (500 M is C, CD3OD): δ 2.54-2.63 (m, 1H), 2.82-3.05 (m, 3H), 3.93-4.15 (m, 2H), 4.66-4.71 (m, 1H), 5.05-5.10 (m, 1H).

Example 12

3-Mercapto-2-[(piperidine-3-carbonyl)-amino]-propionic acid

Specified in the title compound was obtained from piperidine-1,3-dicarboxylic acid 1-tert-butyl ether by the method described in Example 14. Output: 15,1 mg (73%).

1H NMR (500 MHz, CD3OD): δ 1.73-2.10 (m, 4H), 2.84-2.92 (m, 2H), 2.95-3.14 (m, 2H), 3.15-3.29 (m, 3H), 4.56-4.62 (m, 1H).

Example 13

2-[(Azetidin-3-carbonyl)-amino-3-mercapto-propionic acid

Specified in the title compound was obtained from azetidine-1,3-dicarboxylic acid mono-tert-butyl ether by the method described in Example 14. Output: 13.5 mg (71%).

1H NMR (500 MHz, CD3OD): δ 2.86-3.02 (m, 2H), 3.72-3.80 (m, 1H), 4.20-4.24 (d, 4H), 4.62-4.67 (m, 1H).

Example 14

3-(6-Amino-5-methyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

a) 5-Bromo-2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-pyridine

2-Amino-5-bromo-3-methylpyridin (15.0 g, 80.2 mmol) in tert-butanol was treated with di-tert-BUTYLCARBAMATE (43,6 g, 200 mmol) and DMAP (0,60 g, 4,91 mmol). The reaction mixture was left at ambient temperature overnight, and then concentrated under reduced pressure. Added hexane, and the product precipitated as solids. After filtering received 5-bromo-2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-pyridine (22,0 g, 71%).

b) 2-[N,N(tert-Butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-3-methylpyridin

A solution of 5-bromo-2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-pyridine (26,0 g of 67.1 mmol), tert-butyl-dimethyl-tributylstannyl-methoxy-silane (47,6 g, 109 mmol) and bis(triphenylphosphine)palladium(II)dichloride (0,90 g of 1.42 mmol) in 1,2-dichloroethane (80 ml) was stirred at 90°C for two days. The mixture was cooled to 0°and added diethyl ether (200 ml)and then saturated aqueous potassium fluoride (40 ml). The mixture was intensively stirred for 30 minutes and filtered. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 100:0 to 95:5) was obtained 2-[N,N-bis(tert-butoxy-carbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-3-methylpyridin (18.0 g, 59%).

a) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-hydroxymethyl-3-methyl-pyridine.

Tetrabutylammonium fluoride (25,1 g of 79.6 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy-methyl)-3-methylpyridine (18.0 g, 39.8 mmol) in THF (150 ml). The reaction mixture was stirred over night at room temperature. After concentration under reduced pressure and subsequent flash chromatography (hexane/EtOAc, 50:50) was obtained 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-hydroxymethyl-3-methylpyridine (8.0 g, 59%).

d) 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methyl-pyridine

Triphenylphosphine (7,43 g, 28.3 mmol) and GVS 4(9,49 g, 28.6 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-hydroxymethyl-3-methylpyridine (8.00 g, 23.6 mmol) in dichloromethane (220 ml) at 0°C. the Reaction mixture was stirred for 3 hours and then concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 80:20) received 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-3-methylpyridin (8.0 g, 77%).

d) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-5-methyl-pyridin-3-yl-methyl)-malonic acid diethyl ester

To a suspension of NaH (0.24 g, 6.0 mmol, 60%) in DMF (5 ml) was added diethylmalonate (of 0.91 ml, 6.0 mmol)and the mixture was stirred for 15 minutes. Solution was added 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl bromide-3-methyl-pyridine (2.0 g, 5.0 mmol) in DMF (5 ml)and the resulting solution was stirred for 120 minutes at 60°C. was Added ethyl acetate, and this mixture is washed with water and brine and dried. After evaporation of the solvent the crude product was purified flash chromatography (CH3HE/CH2CL2that 1:100 1:20) to give 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (1.2 g, 50%).

e) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-5-methyl-pyridin-3-yl-methyl)-malonic acid monotropy ether

A solution of KOH (154 mg, of 2.75 mmol) in ethanol (2 ml) was added to a solution of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-meth is l-pyridine-3-ylmethyl)-malonic acid diethyl ester (1.2 g, of 2.50 mmol) in ethanol (10 ml) and methylene chloride (4 ml) at 0°C. the Mixture was stirred for 18 hours at room temperature. The mixture was concentrated under reduced pressure, and the residue was dissolved in water. Added ethyl acetate, and the organic layer was washed with 0.5 M HCl, water, brine and dried. After filtration and concentration under reduced pressure, the obtained crude 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-malonic acid monotropy ester (1.0 g, 88%).

W) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-5-methyl-pyridin-3-yl-methyl)-acrylic acid ethyl ester

Diethylamine (0.26 g, to 2.67 mmol) was added to a mixture of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-malonic acid of monoethylene ester (1.0 g, 2.2 mmol) and 37% aqueous formaldehyde solution (0.24 g, 3.00 mmol) in methylene chloride (2 ml) at 0°C. the Mixture was stirred for 16 hours at room temperature was added ethyl acetate. The organic layer was washed with water and 5% NaHCO3and dried. After concentration under reduced pressure and subsequent flash chromatography (toluene/ethyl acetate, 3:1 1:2) was obtained 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0.68 g, 73%).

C) 2-Acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)-amino]-5-methyl-pyridin-3-yl)-propionic acid Adilov the th ether

The triethylamine (0,234 ml, 1,68 mmol) was added to a solution of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0.68 g, of 1.61 mmol) in teoksessa acid (3 ml) at 0°C. the Mixture was stirred at room temperature for 16 hours. Added ethyl acetate, and the organic phase is washed with water, saturated NaHCO3and brine and dried. The crude product was purified flash chromatography (toluene/ethyl acetate, 3:1 1:2) to give pure 2-acetyl-sulfanilyl-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-propionic acid ethyl ester (489 mg, 61%) and 2-acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridine-3-yl)-propionic acid ethyl ester with a small amount of impurities (0.34 g, 43%).

3-(6-Amino-5-methyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

2-Acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-propionic acid ethyl ester (17 mg, 0,034 mmol) was dissolved in concentrated HCl (3.0 ml). The solution was heated under reflux for 1 hour. After concentration under reduced pressure has been specified in the header of the compound (8,9 mg, 100%) in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, CD3OD): δ 2.26 (s, 3H), 2.72-2.75 (m, 2H), 2.83-2.91 (m, 3H), 7.60 (s, 1H), 7.77 (s, 1H).

MS (+) 227 (M+1).

Example 15

3-(6-Amino-4-methyl-pyridine-3-yl)-2-mercaptomethyl-propionic acid

a) 2-Amino-5-bromo-4-methylpyridin

2-Amino-4-methylpyridin (110 g, of 1.02 mol) in Hydrobromic acid (1 l, 48%) was stirred at 70°and was added dropwise a solution of hydrogen peroxide (300 ml, 15%) for one hour at such a speed that the temperature of the reaction mixture remained 70-80°C. the Mixture was stirred for 90 minutes at 70°and poured on crushed ice. the pH is brought to a value of 4-5 with sodium carbonate, and precipitated precipitated solid (containing mainly dibromononane products) was filtered and discarded. Then the pH was raised to 9, and loose precipitated product was collected by filtration. After recrystallization from toluene obtained 2-amino-5-bromo-4-methylpyridin (76,3 g, 40%).

b) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-bromo-4-methylpyridin

2-Amino-5-bromo-4-methylpyridin (5,70 g of 30.5 mmol) in chloroform was treated with di-tert-BUTYLCARBAMATE (20,0 g, 91,60 mmol) and DMAP (0,60 g, 4,91 mmol). The reaction mixture was left to stand at ambient temperature overnight, and then concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 95:5) was obtained 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-bromo 4-methylpyridin (8,02 g, 68%).

a) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-4-methylpyridin

A solution of 2-[N,N-b is(tert-butoxycarbonyl)amino]-5-bromo-4-methyl-pyridine (15.0 g, 38,70 mmol), tert-butyl-dimethyl-tributylstannyl-silane (25.4 g, 58.3 mmol) and bis(triphenylphosphine)palladium(II)dichloride (0,90 g of 1.42 mmol) in 1,2-dichloroethane (50 ml) was stirred at 90°C for two days. This mixture was cooled to 0°and added diethyl ether (200 ml)and then saturated aqueous potassium fluoride (40 ml). The mixture was intensively stirred for 30 minutes and filtered. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 95:5) was obtained 2-[N,N-bis(tert-butoxy-carbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-4-methylpyridine (10.0 g, 57%).

g) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-hydroxymethyl-4-methyl-pyridine

Tetrabutylammonium fluoride (13,9 g of 44.1 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-4-methylpyridine (10.0 g, a 24.3 mmol) in THF (100 ml). The reaction mixture was stirred for 3 hours at room temperature. After concentration under reduced pressure and subsequent flash chromatography (hexane/EtOAc, 50:50) was obtained 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-hydroxymethyl-4-methylpyridine (5.0 g, 67%).

d) 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine

Triphenylphosphine (4,69 g of 17.9 mmol) and GVS4(4,89 g of 14.8 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonylamino]-5-hydroxymethyl-4-methylpyridine (5,00 g, to 22.0 mmol) in dichloromethane (130 ml) at 0°C. the Reaction mixture was stirred for 3 hours and then was diluted with dichloromethane. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 80:20) received 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-4-methylpyridin (5.35 g, 90%).

e) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-4-methyl-pyridine-3-yl-methyl)-malonic acid diethyl ester

To a suspension of NaH (0.24 g, 6.0 mmol, 60%) in DMF (5 ml) was added diethylmalonate (of 0.91 ml, 6.0 mmol)and the mixture was stirred for 15 minutes. Solution was added 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl bromide-4-methyl-pyridine (2.0 g, 5.0 mmol) in DMF (5 ml)and the resulting solution was stirred for 120 minutes at 60°C. was Added ethyl acetate, and this mixture is washed with water and brine, and was dried. After evaporation of the solvent the crude product was purified flash chromatography (CH3HE/CH2CL2that 1:100 1:20) to give pure fractions of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (1,15 g, 48%) and the contaminated fraction 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (1.1 g).

W) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-4-methyl-pyridine-3-yl-methyl)-malonic acid monotropy ether

RAS is a thief CON (141 mg, 2,52 mmol) in ethanol (2 ml) was added to a solution of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (1.1 g, to 2.29 mmol) in ethanol (10 ml) and methylene chloride (4 ml) at 0°C. the Mixture was stirred for 18 hours at room temperature. The mixture was concentrated under reduced pressure, and the residue was dissolved in water. Added ethyl acetate, and the organic layer was washed with 0.5 M HCl, water, brine and dried. After filtration and concentration under reduced pressure, the obtained crude 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-malonic acid monotropy ester (1.0 g, 97%).

C) 2-(6-[N,N-bis(tert-Butoxycarbonyl)amino]-4-methyl-pyridine-3-yl-methyl)-acrylic acid ethyl ester

Diethylamine (0.26 g, to 2.67 mmol) was added to a mixture of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-malonic acid of monoethylene ester (1.0 g, 2.2 mmol) and 37% formaldehyde solution (0.24 g, 3.00 mmol) in methylene chloride (2 ml) at 0°C. the mixture was stirred for 16 hours at room temperature was added ethyl acetate. The organic layer was washed with water and 5% NaHCO3and dried. After concentration under reduced pressure and subsequent flash chromatography (toluene/ethyl acetate, 3:1→1:1) was obtained 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-acrylic is th acid ethyl ester (0,81 g, 88%).

and) 2-Acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)-amino]-4-methyl-pyridine-3-yl)-propionic acid ethyl ester

The triethylamine (0,279 ml, 2.0 mmol) was added to a solution of 2-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0.8 g, 1.9 mmol) in teoksessa acid (3 ml) at 0°C. the Mixture was stirred at room temperature for 16 hours. Added ethyl acetate, and the organic phase is washed with water, saturated Panso3and brine and dried. The crude product was purified flash chromatography (toluene/ethyl acetate, 3:1→1:2) to give pure 2-acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-5-methyl-pyridin-3-yl)-propionic acid ethyl ester (200 mg, 21%) and slightly polluted 2-acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-yl)-propionic acid ethyl ester (0.68 g).

K) 3-(6-Amino-4-methyl-pyridine-3-yl)-2-mercaptomethyl-propionic acid

2-Acetylsalicylate-3-(6-[N,N-bis(tert-butoxycarbonyl)amino]-4-methyl-pyridine-3-yl)-propionic acid ethyl ester (36 mg, 0,072 mmol) was dissolved in concentrated HCl (3.0 ml). The solution was heated under reflux for 1 hour. After concentration under reduced pressure has been specified in the title compound (18.7 mg, 98%) in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, CD3OD): δ 2.42 (s, 3H), 2.72-2.95 (m, 5H), 6.81 (s, 1H), 7.58 (s, 1H).

MS (+) 227 (M+1).

Example 16

2-Mercaptomethyl-3-piperidine-4-yl-butyric acid

a) 4-Formyl-piperidine-1-carboxylic acid tert-butyl methyl ether

Periodinane (26,9 g, 63.5 mmol) was added to a solution of 1-tert-butoxycarbonyl-piperidine-4-methanol (10.5 g, 48.8 mmol) in methylene chloride (200 ml)and the mixture was stirred for 90 minutes. Added diethyl ether and the precipitate was removed by extraction with a mixture of 10% PA2S2O3/saturated NaHCO3(1:1, 300 ml). The organic layer was washed with 0.5 M NaOH and brine, dried and concentrated under reduced pressure. After flash chromatography (hexane/EtOAc, 8:2) was 4-formyl-piperidine-1-carboxylic acid tert-butyl ester (8.5 g, 81%).

b) 2-(1-tert-Butoxycarbonyl-piperidine-4-ylmethylene)-malonic acid diethyl ester

To a solution of diethylmalonate (710 μl, 4.7 mmol) and 4-formyl-piperidine-1-carboxylic acid tert-butyl ester (1.0 g, 4.7 mmol) in methylene chloride (5 ml) was added piperidine (46 μl, 0.47 mmol) and acetic acid (27 μl, 0.47 mmol). The reaction mixture was stirred for 72 hours at room temperature and then for 16 hours at 45°C. was Added EtOAc, and the mixture is washed with water and brine, dried and concentrated under reduced pressure. The crude substance was purified flash chromatog what afia (heptane/EtOAc, 3:1 1:3) to give 2-(1-tert-butoxycarbonyl-piperidine-4-ylmethylene)-malonic acid diethyl ester (0,69 g, 40%).

a) 2-[1-(1-tert-Butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid diethyl ester

MeLi (5,34 ml, 8,54 mmol, 1.6 M in diethyl ether) was added dropwise to a suspension of Cul (0.74 g, 3.88 mmol) in THF (5 ml) at -78°C in an atmosphere of argon, and the mixture was stirred for 30 minutes. A solution of 2-(1-tert-butoxycarbonyl-piperidine-4-ylmethylene)-malonic acid diethyl ester (0,69 g, 1.94 mmol) in THF (5 ml) was added dropwise, and the reaction mixture was stirred for 120 minutes at -78°and then left to warm to room temperature over 60 minutes. Added concentrated aqueous NH4HE, and this mixture then was extracted with EtOAc, washed with concentrated aqueous NH4HE and brine, dried and concentrated under reduced pressure. The crude substance was purified flash chromatography (heptane/EtOAc, 3:1 1:6) to give 2-[1-(1-tert-butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid diethyl ester (0.39 g, 54%).

g) 2-[1-(1-tert-Butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid monotropy ether

A solution of KOH (84 mg, of 1.16 mmol) in tO (2 ml) was added dropwise to a solution of 2-[1-(1-tert-butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid diethyl ester (0.39 g, 1.01 mmol) in methylene chloride (4 ml) and the tO (10 ml) at 0° C. the resulting mixture was stirred at room temperature overnight. Added EtOAc, and the mixture was washed with 0.5 M HCl and brine, dried and concentrated under reduced pressure to obtain 416 mg of the crude 2-[1-(1-tert-butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid of monoethylene ether.

d) 4-(2-Etoxycarbonyl-1-methyl-allyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

Formaldehyde (132 mg, of 1.65 mmol, 37% in water) was added to a solution of the crude 2-[1-(1-tert-butoxycarbonyl-piperidine-4-yl)-ethyl]-malonic acid of monoethylene ester (416 mg) in methylene chloride (2 ml) at 0°C. was Added dropwise diethylamine (153 μl, about 1.47 mmol), and the mixture was stirred at room temperature overnight. Added EtOAc, and the mixture is washed with water and saturated NaHCO3, was dried and concentrated under reduced pressure. The crude substance was purified flash chromatography (toluene/EtOAc, 3:1) to obtain 4-(2-etoxycarbonyl-1-methyl-allyl)-piperidine-1-carboxylic acid tert-butyl ester (0.18 g, yield 49% over two stages).

e) 4-(3-Acetylmethadol-2-etoxycarbonyl-1-methyl-propyl)-piperidine-1-carboxylic acid tert-butyl methyl ether

TEA (86 μl, 0,617 mmol) was added to a solution of 4-(2-etoxycarbonyl-1-methyl-allyl)-piperidine-1-carboxylic acid tert-butyl ester (0.18 g, 0.59 mmol) in teoksessa acid (2 ml) at 0°C. On the Le stirring for 6 hours added teoksessa acid (2 ml), and the mixture was stirred at 45°With during the night. Added EtOAc, and the mixture is washed with water, saturated Panso3and brine, dried and concentrated under reduced pressure. The crude substance was purified flash chromatography (toluene/EtOAc, 5:1-1:1) to obtain 4-(3-acetylmethadol-2-etoxycarbonyl-1-methyl-propyl)-piperidine-1-carboxylic acid tert-butyl ether with a small amount of impurities (0.17 g, 75%).

W) 2-Acetylsalicylate-3-piperidine-4-yl-butyric acid ethyl ester

TFA (2 ml) was added to a solution of 4-(3-acetylmethadol-2-etoxycarbonyl-1-methyl-propyl)-piperidine-1-carboxylic acid tert-butyl ester (0.17 g, 0,439 mmol) in methylene chloride (10 ml). The reaction mixture was stirred for 90 minutes and then concentrated under reduced pressure. The crude product was purified using HPLC (10→50% acetonitrile in water with 0.1% TFA) to give 2-acetylsalicylate-3-piperidine-4-yl-butyric acid ethyl ester (101 mg, 54%) as a TFA salt.

C) 2-Mercaptomethyl-3-piperidine-4-yl-butyric acid

Concentrated hydrochloric acid (4 ml) was added TFA salt of 2-acetylsalicylate-3-piperidine-4-yl-butyric acid ethyl ester (0,101 g, 0,252 mmol) in argon atmosphere. The reaction mixture was heated under reflux for 5 hours, and then concentrated under reduced pressure to obtain vast remerol mixture specified in the connection header (73,7 mg) in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, CD3OD) for the main diastereoisomer: δ 1.10 (d, 3H), 1.36-1.58 (m, 2H), 1.71-1.78 (m, 2H), 1.93-1.99 (m, 1H), 2.05-2.11 (m, 1H), 2.60-2.66 (m, 2H), 2.80-2.86 (m, 1H), 2.94-3.02 (m, 2H), 3.38-3.45 (m, 2H).

MS (+) 218 (M+1).

Example 17

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-butyric acid

a) (5-Formyl-pyridin-2-yl)-carbamino acid tert-butyl methyl ether

(5-Hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ether (7,00 g, and 31.2 mmol) was dissolved in dry SO (50 ml)and the reaction flask was immersed in a water bath at 15°C. was Added TEA (13.1 ml, 94,0 mmol), then portions were added to the complex of sulfur trioxide with pyridine (15.0 g, 94,0 mmol). The reaction mixture was stirred for another 45 minutes and then poured into crushed ice. The product was extracted with diethyl ether, and the combined organic extracts were washed with brine, dried and concentrated under reduced pressure. After recrystallization from a mixture of hexane/CH2Cl2received (5-formyl-pyridin-2-yl)-carbamino acid tert-butyl ether (5,40 g, 78%) as white crystals.

b) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester.

To a solution of diethylmalonate (710 μl, 4.7 mmol) and (5-formyl-pyridin-2-yl)-carbamino acid tert-butyl ester (1.04 g, 4.7 mmol) in a mixture of methylene chloride/DMF (1:1, 5 ml) was added piperidine (46 μl, 0.47 mmol) and acetic acid (2 μl, 0.47 mmol). The reaction mixture was stirred for 72 hours at room temperature and then for 16 hours at 45°C. was Slowly added heptane to obtain 2-(6-tert-butoxycarbonyl-amino-pyridine-3-ylmethylene)-malonic acid diethyl ester (0,69 g, 40%) as gray crystals.

a) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid diethyl ester

MeLi (6.5 ml, 10.4 mmol, 1.6 M in diethyl ether) was added dropwise to a suspension Cul (0.9 g, to 4.73 mmol) in THF (28 ml) at -78°C in argon atmosphere. The reaction mixture was stirred for 30 minutes. A solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester (0.84 g, 2.3 mmol) in THF (7 ml) was added dropwise, and the reaction mixture was stirred for 180 minutes at -78°C. was Added dropwise a saturated aqueous NH4OH, and this mixture was extracted with EtOAc. The organic phase was washed with saturated aqueous NH4OH and NaCl, dried and concentrated under reduced pressure. After flash chromatography (toluene/EtOAc, 3:1 1:6) was obtained 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid diethyl ester (0.723 g, 82,4%) as a white solid.

g) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid monotropy ether

A solution of KOH (113,6 mg, 2.04 mmol) in EtOH (2 ml) was added dropwise to a solution of 2-[1-(6-t is em-butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid diethyl ester (0.7 g, of 1.84 mmol) in methylene chloride (4 ml) and tO (10 ml) at 0°in argon atmosphere, and this reaction mixture was stirred over night. Added 1 M KOH (100 ml)and the mixture was washed with methylene chloride. The aqueous phase was acidified to pH 2 using 2 M HCl, and was extracted with EtOAc. The organic phase was dried and concentrated under reduced pressure to obtain crude 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid of monoethylene ester (423 mg, 65%).

d) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-ethyl]-acrylic acid ethyl ester

Diethylamine (0,153 ml, 1,473 mmol) was added to a solution of 2-[1 -(6-tert-butoxycarbonylamino-pyridine-3-yl)-ethyl]-malonic acid of monoethylene ester (423 mg, 1.2 mmol) and formaldehyde (132 mg, 1,626 mmol, 36% in water) in methylene chloride (2 ml) at 0°C in argon atmosphere. The mixture was stirred at room temperature overnight. Added EtOAc, and the solution was washed with water, NaHCO3and brine, dried and concentrated under reduced pressure. After flash chromatography (toluene/EtOAc, 3:1) was obtained 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-ethyl]-acrylic acid ethyl ester (158 mg, 41%).

e) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-butyric acid ethyl ester

TEA (0,076 ml, 0,542 mmol) was added to a solution of 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-ethyl]-AK the sludge acid ethyl ester (158 mg, 0,493 mmol) in teoksessa acid (2 ml) at 0°C in argon atmosphere. The mixture was stirred at 45°With during the night. Added EtOAc and the solution washed with NaHCO3and brine, dried and concentrated under reduced pressure. After flash chromatography (toluene/EtOAc, 5:1→1:1) received 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-butyric acid ethyl ester with a small amount of impurities (178 mg, 91%).

W) 2-Acetylsalicylate-3-(6-amino-pyridin-3-yl)-butyric acid ethyl ester

TFA (2 ml) was added to a solution of 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-butyric acid ethyl ester (178 mg, 0,449 mmol) in methylene chloride (2 ml). The mixture was stirred for 60 minutes and then concentrated under reduced pressure. After flash chromatography (toluene/EtOAc, 1:6) received not pure 2-acetylsalicylate-3-(6-amino-pyridin-3-yl)-butyric acid ethyl ester (176 mg, 95%). After additional purification HPLC (10→70% acetonitrile in water with 0.1% TFA) received 2-acetylsalicylate-3-(6-amino-pyridin-3-yl)-butyric acid ethyl ester (104 mg, 56%) as a TFA salt.

C) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-butyric acid

Concentrated hydrochloric acid (4 ml) was added to 2-acetylsalicylate-3-(6-amino-pyridin-3-yl)-butyric acid ethyl ether (104 mg, 0,253 mmol) in the atmosphere arg is on. The reaction mixture was heated under reflux for 5 hours, and then concentrated under reduced pressure to get diastereomeric mixture specified in the title compound (61 mg, 92%) as cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O) for the main diastereoisomer: δ 1.26 (d, 3H), 2.49-2.53 (m, 2H), 2.64-2.77 (m, 1H), 2.95-3.02 (m, 1H), 7.02 (d, 1H), 7.69 (d, 1H), 7.88 (m, 1H).

MS (+) 227 (M+1).

Example 18

3-(6-Amino-2-methyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

a) (5-Bromo-6-methyl-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

5-Bromo-6-methyl-pyridin-2-ylamine (25,0 g 133,7 mmol) in a mixture of THF/tert-butanol (1:10, 550 ml) was treated with di-tert-BUTYLCARBAMATE (39,3 g, 180,0 mmol) and DMAP (2,40 g and 19.6 mmol). The reaction mixture was stirred for 4 hours at 40°and concentrated under reduced pressure. After flash chromatography (methylene chloride) received (5-bromo-6-methyl-pyridine-2-yl)-carbamino acid tert-butyl ester (17.0 g, 44%).

b) [5-tert-Butyl-dimethyl-silyloxy)-6-methyl-pyridine-2-yl]-carbamino acid tert-butyl methyl ether

A solution of (5-bromo-6-methyl-pyridine-2-yl)-carbamino acid tert-butyl ester (27.5 g, the 95.8 mmol), tert-butyl-dimethyl-tributylstannyl-ethoxysilane (43,5 g of 100.2 mmol) and bis(triphenylphosphine)palladium(II)dichloride (1,00 g of 1.40 mmol) in 1,2-dichloroethane (350 ml) was stirred under heating with about the military refrigerator for 48 hours. Additionally, bis(triphenylphosphine)palladium(II)dichloride (1,00 g of 1.40 mmol) was added every 12 hours. The mixture was cooled to 0°and added diethyl ether (300 ml)and then saturated aqueous potassium fluoride (100 ml). The mixture was intensively stirred for 60 minutes and filtered. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (MeOH/CH2Cl2, 1:99) received 5-(tert-butyl-dimethyl-silyloxy)-6-methyl-pyridine-2-yl]-carbamino acid tert-butyl ester (15 g, 47%).

in) (5-Hydroxymethyl-6-methyl-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

Tetrabutylammonium fluoride (19,6 g of 62.4 mmol) was added to a solution of 5-(tert-butyl-dimethyl-silyloxy)-6-methyl-pyridine-2-yl]-carbamino acid tert-butyl ester (10.5 g, 31,23 mmol) in THF (100 ml) and stirred at room temperature overnight. Was added water, and the product was concentrated by chloroform. The organic phase was dried and concentrated under reduced pressure. After flash chromatography (MeOH/CH2Cl2, 2,5:77,5) received 5-hydroxymethyl-6-methyl-pyridine-2-yl)-carbamino acid tert-butyl ester (6.0 g, 81%).

d) 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine

Triphenylphosphine (9,83 g, 37.5 mmol) and GVS4(of 17.7 G. of 53.5 mmol) was added to a solution of (5-hydroxymethyl-6-methyl-pyridine-yl)-carbamino acid tert-butyl ether (8,50 g, 35,7 mmol) in dichloromethane (30 ml) at 0°C. the Reaction mixture was stirred for 3 hours at room temperature, and then washed with dichloromethane. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (CH2CL2received 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine (of 4.05 g, 38%).

d) 2-(6-tert-Butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester.

A solution of diethylmalonate (1,21 ml of 7.97 mmol) in DMF (2 ml) was added dropwise to a suspension of NaH (348 mg, of 7.97 mmol, 55% in mineral oil) in DMF (5 ml) at 0°C in argon atmosphere. The reaction mixture was stirred for 45 minutes and was added dropwise a solution of (5-methyl bromide-6-methyl-pyridine-2-yl)carbamino acid tert-butyl ester (2.0 g, only 6.64 mmol) in DMF (5 ml). The mixture was stirred overnight (0°-20°). Added EtOAc, and the solution was washed with water and brine, dried and concentrated under reduced pressure. After flash chromatography (heptane/EtOAc, 4:1) was obtained 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (1,87 g, 74%).

e) 2-(6-tert-Butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-malonic acid monotropy ether

A solution of KOH (300 mg, to 5.35 mmol) in tO (4 ml) was added to a solution of 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-metil)-malonic acid diethyl ester (1.85 g, a 4.86 mmol) in a mixture tO/methylene chloride (2:1, 21 ml) at 0°C. the Mixture was stirred for 40 hours at room temperature and was added EtOAc. The mixture was washed with 0.5 M HCl and brine, dried and concentrated under reduced pressure to obtain crude 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-malonic acid of monoethylene ester (1.45 g).

W) 2-(6-tert-Butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester

Diethylamine (359 mg, of 4.90 mmol) was added dropwise to a solution of 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-malonic acid of monoethylene ester (1.44 g, 4.09 to mmol) and formaldehyde (464 mg, 5,72 mmol, 37% in water) in methylene chloride (35 ml) at 0°C in argon atmosphere. The mixture was stirred at room temperature overnight. Added methylene chloride, and the solution was washed Na2CO3and brine, dried and concentrated under reduced pressure to obtain 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (1,03 g, 79%).

C) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-2-methyl-pyridin-3-yl)-propionic acid ethyl ester

TEA (0,556 ml of 3.99 mmol) was added to a solution of 2-(6-tert-butoxycarbonylamino-2-methyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (1.23 g, of 3.84 mmol) in teoksessa acid (10 ml) at 0°in which atmosphere argon. The mixture was stirred at room temperature for 64 hours. Added EtOAc, and the solution was washed Na2CO3and brine, dried and concentrated under reduced pressure. After flash chromatography (toluene/EtOAc, 5:1→1:1) received 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-2-methyl-pyridin-3-yl)-propionic acid ethyl ester (1,33 g, 87%).

3-(6-Amino-2-methyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

Concentrated hydrochloric acid (2 ml) was added to 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-2-methyl-pyridin-3-yl)-propionic acid ethyl ether (77 mg, 0,19 mmol) in argon atmosphere. The reaction mixture was heated under reflux for 110 minutes and then concentrated under reduced pressure to obtain specified in the title compound (39 mg, 76%) as cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 2.49 (s, 3H), 2.73-2.92 (m, 5H), 6.81 (d, 1H), 7.77 (d, 1H).

MS (+) 227 (M+1).

Example 19

2-Acetylsalicylate-3-(2-amino-pyrimidine-5-yl)-propionic acid ethyl ester

a) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-bromopyrimidine

2-Amino-6-bromopyrimidine (9.00 g, of 51.7 mmol) in a mixture of THF/tert-butanol (1:1, 100 ml) was treated with di-tert-BUTYLCARBAMATE (34,0 g, 156,0 mmol) and DMAP (3.00 g, 24.5 mmol). The reaction mixture was left to stand at ambient temperature for the eyes and concentrated under reduced pressure. The residue was distributed between dichloromethane and water and brought the pH to 4 1 M HCl. The solution was extracted with dichloromethane, dried and concentrated under reduced pressure. The crude product is suspended in hexane and filtered to obtain 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-bromopyrimidine (15.0 g, 77%).

b) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-pyrimidine

A solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-bromopyrimidine (16.0 g, 45,0 mmol), tert-butyl-dimethyl-tributylstannyl-silane (20,5 g of 47.1 mmol) and bis(triphenylphosphine)palladium(II)dichloride (1,00 g of 1.40 mmol) in 1,2-dichloroethane (50 ml) was stirred while boiling under reflux overnight. Added addition of bis(triphenylphosphine)palladium(II)dichloride (1,00 g of 1.40 mmol)and the solution boiled under reflux for 8 hours. The mixture was cooled to 0°and was added diethyl ether (200 ml), then saturated aqueous potassium fluoride (50 ml). The mixture was intensively stirred for 60 minutes and filtered. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (MeOH/CH2Cl2, 1:99) received 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy)-pyrimidine (10.2 g, 55%).

a) 2-[N,N-bis(tert-Butoxycarbonyl)amino]-5-hydroxymethyl-pyrimidine

p> Tetrabutylammonium fluoride (15.3 g, to 48.6 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-(tert-butyl-dimethyl-silyloxy-methyl)-pyrimidine (10.0 g, a 24.3 mmol) in THF (100 ml) and stirred at room temperature overnight. Was added water, and the product was extracted with chloroform. The organic phase was dried and concentrated under reduced pressure. After flash chromatography (MeOH/CH2Cl2, 2,5:77,5) received 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-hydroxymethyl-pyrimidine (4,20 g, 53%).

d) 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine

Triphenylphosphine (2,71 g of 10.73 mmol) and GVS4(4,89 g of 14.8 mmol) was added to a solution of 2-[N,N-bis(tert-butoxycarbonyl)amino]-5-hydroxymethyl-pyrimidine (3,20 g, 9,83 mmol) in dichloromethane (30 ml) at 0°C. the Reaction mixture was stirred for 1 hour and then was diluted with dichloromethane. The organic phase is washed with water, dried and concentrated under reduced pressure. After flash chromatography (CH2Cl2) received 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine (2.55 g, 67%).

d) 2-(2-[N,N-bis(tert-Butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid diethyl ester

A solution of diethylmalonate (0,704 ml, with 4.64 mmol) in DMF (2 ml) was added dropwise to a suspension of NaH (200 mg, with 4.64 mmol, 55% in mineral oil) in DMF (4 ml) at 0°C in argon atmosphere. Reactio the ing the mixture was stirred for 30 minutes and was added dropwise a solution of 5-methyl bromide-2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine (1.5 g, 3,86 mmol) in DMF (4 ml). The mixture was stirred at room temperature for 3 hours. Added EtOAc, and the mixture is washed with water and brine, dried and concentrated under reduced pressure. After flash chromatography (heptane/EtOAc, 3:2) was obtained 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid diethyl ester (0.87 g, 40%).

e) 2-(2-[N,N-bis(tert-Butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid monotropy ether

A solution of KOH (106 mg, 1.88 mmol) in EtOH (2 ml) was added to a solution of 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid diethyl ester (0,80 g, 1,71 mmol) in a mixture of EtOH/methylene chloride (2:1, 12 ml) at 0°C. the Mixture was stirred for 16 hours at room temperature and was added EtOAc. The mixture was washed with 0.5 M HCl and brine, dried and concentrated under reduced pressure to obtain 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid of monoethylene ether (0,67 g, 89%).

W) 2-(2-[N,N-bis(tert-Butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-acrylic acid ethyl ester

Diethylamine (0.124 g, was 1.69 mmol) was added dropwise to a solution of 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-malonic acid of monoethylene ether (0,62 g of 1.41 mmol) and formaldehyde (160 mg, 2.0 mmol, 37% in water) in methylene chloride (15 ml) at 0°C in argon atmosphere. The mixture is eremetical at room temperature over night. Added EtOAc, and the solution was washed with water, Panso3and brine, dried and concentrated under reduced pressure to obtain crude 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-acrylic acid ethyl ester (0.54 g, 94%).

C) 2-Acetylsalicylate-3-(2-[N,N-bis(tert-butoxycarbonyl)-amino]-pyrimidine-5-yl)-propionic acid ethyl ester

TEA (0,189 ml, 1.35 mmol) was added to a solution of 2-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-ylmethyl)-acrylic acid ethyl ester (0,53 g of 1.30 mmol) in teoksessa acid (13 ml) at 0°C in argon atmosphere. The mixture was stirred at room temperature for 40 hours. Added EtOAc, and the mixture was washed PA2CO3and brine, dried and concentrated under reduced pressure. After flash chromatography (heptane/EtOAc, 5:1→4:1) received 2-acetylsalicylate-3-(2-[N,N-bis-(tert-butoxycarbonyl)amino]-pyrimidine-5-yl)-propionic acid ethyl ester (0.56 g, 89%).

and) 2-Acetylsalicylate-3-(2-amino-pyrimidine-5-yl)-propionic acid ethyl ester

TFA (1.5 ml) was added to a solution of 2-acetylsalicylate-3-(2-[N,N-bis(tert-butoxycarbonyl)amino]-pyrimidine-5-yl)-propionic acid ethyl ester (225 mg, 0.46 mmol) in methylene chloride (1.5 ml). The reaction mixture was stirred for 120 minutes and then concentrated under reduced pressure to receive the receiving specified in the title compound (172 mg, 94%) as a TFA salt.

1H NMR (500 MHz, CD3OD): δ 1.22 (t, 3H), 2.35 (s, 3H), 2.80-3.00 (m, 3H), 3.00-3.22 (m, 2H), 4.13 (q, 2H), 8.43 (s, 2H).

Example 20

2-(6-Amino-pyrimidine-3-ylmethyl)-3-mercapto-butyric acid

a) 3-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-(diethoxy-phosphoryl)-propionic acid ethyl ester

To a suspension of NaH (1,17 g, 60% in mineral oil, of 29.3 mmol) in DMF (70 ml) was added triethylphosphite (6,01 g, 26,82 mmol) at 0°C. the Reaction mixture was allowed to mix at 0°C for 15 minutes. To the reaction mixture were added 5-methyl bromide-pyridine-2-yl)-carbamino acid tert-butyl ester (7.0 g, 24,38 mmol) at 0°and this reaction mixture was stirred at room temperature for 16 hours. The reaction was suppressed by slow addition of saturated aqueous ammonium chloride (70 ml). The mixture was extracted with EtOAc, washed with brine and dried. The crude product was purified column chromatography (CH2Cl2/EtOAc, 1:0 1:1 0:1) with 3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-(diethoxy-phosphoryl)-propionic acid ethyl ester (5.1 g, 50%).

b) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-but-2-ene acid ethyl ester

To a suspension of NaH (much as 278.8 mg, 60% in mineral oil, 6,97 mmol) in THF (25 ml) at 0°C was added a solution of 3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-(diethoxy-phosphoryl)-propionic sour the s ethyl ester (2.5 g, of 5.81 mmol) in THF (30 ml). The reaction mixture was allowed to mix at 0°C for 1 hour. To the reaction mixture was added acetaldehyde (512 mg, 11.6 mmol) dropwise at 0°C. the Reaction mixture was left to warm to room temperature and then was stirred for 16 hours. In the reaction vessel was added acetaldehyde (2.0 g)and the reaction mixture was allowed to mix at room temperature for another 16 hours. The reaction was suppressed by slow addition of saturated aqueous ammonium chloride (30 ml). The mixture was extracted with EtOAc, washed with brine and was dried to obtain the crude product. The crude product was purified column chromatography (EtOAc/hexane, 1:8) to obtain ethyl 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-but-2-ene acid ethyl ester as a mixture of isomers (1.1 g, 60%).

C) 3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester

To thioglucose acid (15 ml) was added Et3N (1.5 g, of 14.8 mmol) and ethyl 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-but-2-ene acid ethyl ester (920 mg, 2.9 mmol) at room temperature. The reaction mixture was stirred at 40-45°C for 7 days (optional toluxury acid (1.5 ml) was added to the reaction mixture every two days). The reaction mixture was cooled to room temperature is, and then was diluted with EtOAc (50 ml). The organic layer was separated, washed with saturated NaHCO3, brine and dried. The combined organic layers were concentrated under reduced pressure. The crude product was then purified by three chromatographic columns (CH2Cl2, EtOAc/hexane, 1:5 and acetone/hexane, 1:8) to obtain 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester in the form of diastereomeric mixture (780 mg, 68%). Diastereomers the mixture was separated using preparative chiral chromatography, according to the methodology described below, to obtain the four isomers of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/B, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/In and 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/Year

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/G

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/G was isolated from a mixture of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ileti is)-butyric acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxy-carbylamine-pyridine-3-ylmethyl)-butyric acid ethyl ester/B, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/3-acetylmethadol-2-(6-tert-butoxy-carbylamine-pyridine-3-ylmethyl)-butyric acid ethyl ester/G column (CHIRALCEL OJ, elwira mixture isohexane:acetonitrile:isopropyl alcohol:diethylamine(99; 0,5; 0,5; 0,1). According to HPLC using a column CHIRALPACK OJ, eluruumi mixture isohexane:ethanol:diethylamine (99; 1; 0.5), and the enantiomeric excess was > 99%.

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/A

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester And was isolated from a mixture of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxy-carbylamine-pyridine-3-ylmethyl)-butyric acid ethyl ester/B, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/3-acetylmethadol-2-(6-tert-butoxy-carbylamine-pyridine-3-ylmethyl)-butyric acid ethyl ester/G column (CHIRALCEL OJ, elwira mixture isohexane:1-propanol:diethylamine (98; 2; 0,1). According to HPLC using a column CHIRALPACK OJ,eluruumi mixture isohexane:ethanol:diethylamine (99; 1; 0.5), and the enantiomeric excess was > 99%.

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/B

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/B was isolated from a mixture of 3-acetyl-effect-free remedy 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxycarbonyl-amino-pyridine-3-ylmethyl)-butyric acid ethyl ester/B, 3-acetyl-effect-free remedy 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/3-acetylmethadol-2-(6-tert-butoxycarbonyl-amino-pyridine-3-ylmethyl)-butyric acid ethyl ester/G column (CHIRALCEL OJ, elwira mixture isohexane:1-propanol:diethylamine (99; 1; 0,5). According to HPLC using a column CHIRALPACK OJ, eluruumi mixture isohexane:ethanol:diethylamine (99; 1; 0.5), and the enantiomeric excess was > 99%.

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-butyric acid ethyl ester/was isolated from a mixture of 3-acetyl-effect-free remedy 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-oil KIS is the notes of ethyl ether/B, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/G column (CHIRALCEL AS, elwira a mixture of hexane:ethanol:diethylamine (99:1:0,5). According to HPLC using a column CHIRALPACK AS eluruumi mixture isohexane:ethanol:diethylamine (99; 1; 0.5), and the enantiomeric excess was more than 87%.

g) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-butyric acid/A.

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester (45 mg, 0.11 mmol) in concentrated HCl (2 ml) was boiled under reflux in an argon atmosphere for 1 hour. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to obtain 28.4 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (400 MHz, D2O): δ 7.89 (d, 1H), 7.71 (s, 1H), 7.03 (d, 1H), 3.23-3.33 (m, 1H), 3.1-3.2 (m, 1H), 2.75-2.9 (m, 2H), 1.47 (d, 3H).

MS (+) 227 (M+1).

d) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-butyric acid/B

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/B (55 mg, 0.14 mmol) in concentrated HCl (2 ml) was boiled under reflux in an argon atmosphere for 1 hour. Rea is operating and the mixture was left to cool to room temperature and concentrated under reduced pressure to get to 39.4 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (400 MHz, D2O): δ 7.89 (d, 1H), 7.71 (s, 1H), 7.03 (d, 1H), 3.23-3.33 (m, 1H), 3.1-3.2 (m, 1H), 2.75-2.9 (m, 2H), 1.47 (d, 3H).

MS (+) 227 (M+1).

e) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-butyric acid/

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/(9 mg, 0.02 mmol) in concentrated HCl (0.5 ml) was boiled under reflux in an argon atmosphere for 1 hour. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to obtain 6.4 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (400 MHz, D2O): δ 7.82-7.9 (m, 1H), 7.67 (br s, 1H), 7.0 (d, 1H), 3.16-3.28 (m, 1H), 2.96-3.04 (m, 1H), 2.76-2.86 (m, 2H), 1.47 (d, 3H).

MS (+) 227 (M+1).

W) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-butyric acid/G

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-butyric acid ethyl ester/(9 mg, 0.02 mmol) in concentrated HCl (0.5 ml) was boiled under reflux in an argon atmosphere for 1 hour. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to get to 6.8 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (400 MHz, D2O): δ 7.82-7.9 (m, 1H), 7.67 (br s, 1H), 7.0 (d, 1H), 3.16-3.28 (m, 1H), 2.96-3.04 (m, 1H), 2.76-2.86 (m, 2H), 1.47 (d, 3H).

MS (+) 227 (M+1).

Example 21

2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-pentane acid

a) 2-(6-tert-Butoxycarbonylamino-pyridine-3-ylmethyl)-Penta-2-ene acid ethyl ester

To a solution of NaH (290,5 mg, 60% in mineral oil, 7.5 mmol) in THF (25 ml) at 0°C was added a solution of 3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-(diethoxy-phosphoryl)-propionic acid ethyl ester (2.5 g, of 5.81 mmol) in THF (30 ml). The reaction mixture was allowed to mix at 0°C for 1 hour. To the reaction mixture was added Propionaldehyde (725 mg, 12.5 mmol) dropwise at 0°C. the Reaction mixture was allowed to mix at 0°C for 16 hours. Added Propionaldehyde (2.5 g), and the mixture was stirred at room temperature for another 16 hours. The reaction was suppressed by slow addition of saturated NH4Cl (30 ml). The mixture was extracted with EtOAc, washed with brine and dried. The crude product was purified column chromatography (EtOAc/hexane, 1:8) to give 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-Penta-2-ene acid ethyl ester as a mixture of isomers (1.2 g, 60%).

b) 3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester

To thioglucose acid (15 ml) was added Et3N (1.8 g, 17 mmol) and 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-Penta-2-ene acid ethyl EF the R (1.1 g, 3.3 mmol) at room temperature. The reaction mixture was stirred at 65°C for 8 days (optional toluxury acid (1.5 ml) was added to the reaction mixture every two days). The reaction mixture was cooled to room temperature, and then diluted with EtOAc. The organic layer was separated, washed with saturated Panso3, brine and dried. The combined organic layers were concentrated under reduced pressure. The residue is then purified in two chromatographic columns (CH2Cl2and EtOAc/hexane, 1:5) to obtain 350 mg of the mixture of target products and unreacted starting materials. The crude product was further purified HPLC (EtOAc/hexane, 1:9), and then column chromatography (acetone/hexane, 1:8) to obtain specified in the connection header in the form diastereomeric mixture (180 mg, 18%). Diastereomer a mixture of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester were separated using preparative chiral chromatography, according to the methodology described below, to obtain the four isomers of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester/B, 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridi the-3-ylmethyl)-pentanol acid ethyl ester/3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester/Year Diastereomers the mixture was divided into two columns (CHIRALCEL OJ, which were connected with one another, elwira mixture isohexane:isopropyl spirtitual (97:1:2). The enantiomeric excess was determined by analytical HPLC, using two-column CHIRALCEL OJ, which were connected with one another, elwira mixture isohexane:isopropyl alcohol: methanol (97:1:2). 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester And In the enantiomeric excess was more than 99%, and for 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester G he was 97%.

a) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-pentane acid/A.

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester (of 50.4 mg, 0.12 mmol) in concentrated Hcl (2 ml) was boiled under reflux in an argon atmosphere for 1.5 hours. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to get to 33.9 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 7.87 (dd, 1H), 7.67 (d, 1H), 7.0 (d, 1H), 3.02-3.16 (m, 2H), 2.79-2.87 (m, 2H), 1.79-1.88 (m, 1 H), 1.53-1.64 (m, 1 H), 1.07 (t, 3H).

MS (+) 241 (M+1).

g) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-pentane kislota

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester/B (51,6 mg, 0.13 mmol) in concentrated HCl (2 ml) was boiled under reflux in an argon atmosphere for 1.5 hours. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to obtain 34.7 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 7.87 (dd, 1H), 7.67 (d, 1H), 7.0 (d, 1H), 3.02-3.16 (m, 2H), 2.79-2.87 (m, 2H), 1.79-1.88 (m, 1H), 1.53-1.64 (m, 1H), 1.07 (t, 3H).

MS (+) 241 (M+1).

d) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-pentane acid/

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-pentanol acid ethyl ester/(4.3 mg, 0.01 mmol) in concentrated HCl (2 ml) was boiled under reflux in an argon atmosphere for 1.5 hours. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to get to 2.9 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 7.86 (dd, 1H), 7.66 (br s, 1H), 7.0 (d, 1H), 2.94-3.04 (m, 2H), 2.74-2.9 (m, 2H), 1.88-1.97 (m, 1H), 1.55-1.66 (m, 1H), 1.05 (t, 3H).

MS (+) 241 (M+1).

e) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-pentane acid/G

A solution of 3-acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-ilma who yl)-pentanol acid ethyl ester/G (19.2 mg, 0.05 mmol) in concentrated HCl (2 ml) was boiled under reflux in an argon atmosphere for 1.5 hours. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to obtain 12.9 mg specified in the connection header in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 7.86 (dd, 1H), 7.66 (br s, 1H), 7,0 (d, 1H), 2.94-3.04 (m, 2H), 2.74-2.9 (m, 2H), 1.88-1.97 (m, 1H), 1.55-1.66 (m, 1H), 1.05 (t, 3H).

MS (+) 241 (M+1).

Example 22

3-(6-Amino-5-chloro-pyridine-3-yl)-2-mercaptomethyl-propionic acid

a) 6-Amino-5-chloro-nicotinic acid ethyl ester

N-Chlorosuccinimide (21,7 g rate £ 0.162 mol) was added to a suspension of 6-amino-nicotinic acid ethyl ester (18.0 g, to 0.108 mol) in acetonitrile (270 ml), and the mixture is boiled under reflux for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was dissolved in dichloromethane, washed with water and dried. After flash chromatography (2,5%Meon in CH2CL2) received pure 6-amino-5-chloro-nicotinic acid ethyl ester (17.23 g, 79%).

b) 6-bis(tert-Butoxycarbonyl)-amino-5-chloro-nicotinic acid ethyl ester

DMAP (0.11 g, 0.9 mmol) and (BOC)2O (21,54 g, 99 mmol) was added to a solution of 6-amino-5-chloro-nicotinic acid ethyl ester (9.0 g, 45 mmol) in dichloromethane (250 ml). The reaction mixture was stirred shall within 24 hours. DMAP (0.02 equivalent) and (BOC)2O (3×0.5 equivalent) was added during the reaction. The reaction mixture was washed with water and dried. The crude product was washed with hexane to obtain pure 6-bis(tert-butoxycarbonyl)-amino-5-chloro-nicotinic acid ethyl ester (11.87 per g, 66%).

in) (3-chloro-5-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

LiAlH4(2.4 g, 63.2 mmol) was added in portions over a time period of 3.5 hours to a solution of 6-bis(tert-butoxycarbonyl)-amino-5-chloro-nicotinic acid ethyl ester (11.5 g, 28.6 mmol) in THF (70 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight, then carefully added NH4Cl (saturated), then water. The solution was filtered, dried and concentrated under reduced pressure to get crude (3-chloro-5-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ether (5,86 g, 79%).

g) (5-methyl bromide-3-chloro-pyridine-2-yl)-carbamino acid tert-butyl methyl ether

Triphenylphosphine (2,61 g, 9.7 mmol), and then tetrabromide carbon (4,58 g of 13.8 mmol) was added to a suspension of (3-chloro-5-hydroxymethyl-pyridine-2-yl)-carbamino acid tert-butyl ester (2.38 g, 9.2 mmol) in CH2Cl2(60 ml) at 0°C. the Mixture was stirred at room temperature for 5 hours and concentrated under reduced pressure. Added clonicel (40 ml), this mixture was stirred at -20 ° °With during the night. The mixture is then filtered, and the crystalline residue was washed with cold acetonitrile. The filtrate was concentrated under reduced pressure, and another portion of the bromide was obtained as described above. (5-methyl bromide-3-chloro-pyridine-2-yl)-carbamino acid tert-butyl ester (1.86 g, 63%) was obtained as white crystals.

d) 2-(6-tert-Butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-malonic acid diethyl ester

Diethylmalonate (of 1.87 ml, 12.31 mmol) was added to a suspension of NaH (0.54 g, 12,31 mmol, 55%) in dry DMF (15 ml) at -8°C. the mixture was stirred for 15 minutes and then was added dropwise to a solution of (5-methyl bromide-3-chloro-pyridine-2-yl)-carbamino acid tert-butyl ether (3,30 g, 10.26 mmol) in dry DMF (50 ml) at 0°C. the resulting solution was stirred for 40 minutes at 0°and then carefully added NH4Cl (5 ml, saturated). Stirring at room temperature over night and concentration under reduced pressure gave a residue that was dissolved in a mixture of water/CH2CL2. The aqueous layer was extracted with CH2Cl2, and the combined organic extracts were dried, filtered and concentrated under reduced pressure. After flash chromatography (1% Meon in CH2Cl2) received 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)noted that a well known the first acid diethyl ester (2,45 g, 60%) as a sticky transparent oil.

e) 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-malonic acid monotropy ether

A solution of KOH (0,44 g, 6,72 mmol, 85%) in ethanol (5 ml) was added to a solution of 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-malonic acid diethyl ester (2,45 g, 6,11 mmol) in ethanol (25 ml) and methylene chloride (10 ml) at 0°C. the Mixture was stirred for 18 hours at room temperature. The solvent is evaporated in vacuo, and the residue was dissolved in water. The aqueous layer was washed with ether, acidified to pH 4 1 M HCl and was extracted with methylene chloride and ethyl acetate. The combined organic layers were washed with water, brine and dried. After filtration and concentration under reduced pressure, the obtained crude product, which was purified flash chromatography (10%Meon in CH2Cl2obtaining 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-malonic acid of monoethylene ether (1,41 g, 62%) as a yellow-white glassy foam.

W) 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-acrylic acid ethyl ester

To a mixture of 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-malonic acid of monoethylene ester (1.40 g, of 3.64 mmol) and 37% aqueous formaldehyde (0,29 ml, 3.75 mmol) in CH2Cl2(2 ml) at 0°C was added dropwise diethylamine (to 3.67 ml,to 3.67 mmol), and then water (2.5 ml) and CH2Cl2(2.5 ml). The mixture was stirred for 20 hours at room temperature, and then poured into ice water and was extracted with methylene chloride. The organic layer is washed with 5% NaHCO3, was dried and concentrated under reduced pressure. After flash chromatography (1-2,5% Meon in CH2CL3) received 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-acrylic acid ethyl ester 0,81 g (65%).

C) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-propionic acid ethyl ester

Teoksessa acid (4 ml) was added to a suspension of 2-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0,732 g of 2.16 mmol) and triethylamine (0.31 in ml, of 2.23 mmol) at 0°C. the Mixture was stirred at room temperature in an argon atmosphere, poured into ice water and was extracted with CH2Cl2. The organic phase was washed with saturated NaHCO3until the gas evolution stops, and then dried. The crude product was purified twice flash chromatography (CH2Cl2, 1-2,5% Meon in CH2Cl2and hexane/EtOAc, 5:2 1:1) to give pure 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-propionic acid ethyl ester (0,79 g, 87%).

3-(6-Amino-5-chloro-pyridine-3-yl)-2-mercaptomethyl-propionic acid

<> A solution of 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-propionic acid ethyl ester (55 mg, 0,132 mmol) in concentrated HCl (4 ml) was boiled under reflux for 90 minutes. The reaction mixture was cooled and concentrated under reduced pressure to obtain specified in the title compound as HCl salt (36 mg, 96.4 per cent).

1H NMR (400 MHz, D2O): δ 2.70-2.97 (m, 5H), 7.73 (s, 1H), 8.09 (s, 1H).

MS (+) 248 (M+1).

Example 23

3-(6-Amino-5-hydroxymethyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

a) 6-bis(tert-Butoxycarbonyl)amino-5-vinyl-nicotinic acid ethyl ester

A mixture of 5-bromo-6-bis(tert-butoxycarbonyl)amino-nicotinic acid ethyl ester (4,50 g, 10.1 mmol), vinyltrimethylsilane (3,52 g, 11.1 mmol) and tetraacetylethylenediamine (0.50 g, 0.40 mmol) in THF (15 ml) was stirred while boiling under reflux for 24 hours. Added tetracosapentaenoic (0.50 g) and after 24 hours boiling under reflux the reaction mixture was cooled and was diluted with dichloromethane (100 ml). Was added a saturated aqueous KF (25 ml)and the solution was stirred for 1 hour. Was added water, and the product was extracted with dichloromethane, the organic phase was dried and concentrated under reduced pressure. After flash chromatography (1% Meon in dichloromethane) received the 6-bis(tert-butoxycarbonyl)-amino-5-vinyl-nicotinic acid ethyl ester 3,20 g (81%).

b) (5-Hydroxymethyl-3-vinyl-pyridine-2-yl)carbamino acid tert-butyl methyl ether

DIBAL (25 ml, 1 M in hexane) was added dropwise to a solution of 6-bis(tert-butoxycarbonyl)-amino-5-vinyl-nicotinic acid ethyl ester (2.00 g, 5.1 mmol) in THF (40 ml) at 0°C. the Mixture was stirred at room temperature for 1 hour. Caution was added NH4Cl (saturated), and then water, and this mixture was concentrated under reduced pressure. The residue is suspended in 5% Meon in dichloromethane and filtered through silica gel. The filtrate was dried and concentrated under reduced pressure to obtain (5-hydroxymethyl-3-vinyl-pyridine-2-yl)carbamino acid tert-butyl ether (1,00 g, 78%).

in) (5-methyl bromide-3-vinyl-pyridine-2-yl)carbamino acid tert-butyl methyl ether

To a stirred suspension of (5-hydroxymethyl-3-vinyl-pyridine-2-yl)carbamino acid tert-butyl ester (10.0 g, 40.0 mmol) in CH2Cl2(150 ml) and THF (60 ml) was added triphenylphosphine (11.5g, to 44.0 mmol), and then tetrabromide carbon (19.9 g, 60.0 mmol) at 0°C. the Mixture was stirred at room temperature for 1 hour and then concentrated under reduced pressure. Added acetonitrile (100 ml), and the mixture is kept at -20°With during the night. The mixture is then filtered, and the crystalline precipitate was washed with cold acetonitrile. The product was purified flash chrome is cografya (1% Meon in dichloromethane) to give (5-methyl bromide-3-vinyl-pyridine-2-yl)-carbamino acid tert-butyl ester (4.5 g, 36%).

g) 2-(6-tert-Butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid diethyl ester

Diethylmalonate (2.30 g, and 14.3 mmol) was added to a suspension of NaH (0,61 g of 14.3 mmol, 55%) in dry DMF (75 ml) at 0°C. the mixture was stirred for 15 minutes and then was added dropwise to a solution of (5-methyl bromide-Z-chloro-pyridine-2-yl)-carbamino acid tert-butyl ether (4,50 g of 14.3 mmol) in dry DMF (100 ml) at 0°C. the resulting solution was stirred for 40 minutes at 0°and then carefully added NH4Cl (30 ml, saturated). After concentration under reduced pressure, the obtained residue, which was dissolved in a mixture of water/CH2CL2. The aqueous layer was extracted with CH2CL2, and the combined organic extracts were dried, filtered and concentrated under reduced pressure. After flash chromatography (1% Meon in CH2CL2) received 2-(6-tert-butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (4,30 g, 77%).

d) 2-(6-tert-Butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid monotropy ether

A solution of KOH (0.71 g, 12.6 mmol, 85%) in ethanol (10 ml) was added to a solution of 2-(6-tert-butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid diethyl ester (4,30 g, 11.0 mmol) in ethanol (25 ml) and dichloromethane (10 ml) at 0°C. the Mixture was stirred for 6 hours is at room temperature. The solvent is evaporated in vacuo, and the residue was dissolved in water. The aqueous layer was washed with ether, acidified to pH 4 1 M HCl and was extracted with dichloromethane. The organic layer was washed with water and brine and dried. After filtration and concentration under reduced pressure and subsequent flash chromatography (10% Meon in CH2Cl2) received 2-(6-tert-butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid monotropy ether (3,05 g, 76%).

e) 2-(6-tert-Butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester

Diethylamin of 0.85 ml, 8,80 mmol) was added dropwise to a mixture of 2-(6-tert-butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-malonic acid of monoethylene ether (3,05 g of 8.37 mmol) and 37% aqueous formaldehyde (0.71 g, 8,80 mmol) in CH2Cl2(2 ml) at 0°C. the Mixture was stirred for 3 hours at room temperature, and then poured into ice water and was extracted with dichloromethane. The organic layer was washed 55% NaHCO3and dried. After flash chromatography (1% Meon in CH2Cl2) received 2-(6-tert-butoxycarbonylamino-5-vinyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (1.70 g, 61%).

W) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-vinyl-pyridin-3-yl)-propionic acid ethyl ester

Teoksessa acid (4 ml) was added to a suspension of 2-(6-tert-butoxy-is ebonyline-5-vinyl-pyridine-3-ylmethyl)-acrylic acid ethyl ester (0.73 g, of 2.16 mmol) in triethylamine (0.31 in ml, of 2.23 mmol) at 0°C. the Mixture was stirred at room temperature in an argon atmosphere overnight, poured into ice water and was extracted with CH2CL2. The organic phase was washed with saturated NaHCO3until the gas evolution stops, and then dried. After flash chromatography (1% Meon in CH2Cl2) received a 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-chloro-pyridine-3-ylmethyl)-propionic acid ethyl ester (0.51 g, 70%).

C) 3-(6-tert-Butoxycarbonylamino-5-hydroxymethyl-pyridin-3-yl)-mercapto-methyl-propionic acid ethyl ester

Ozone was barbotirovany through a solution of 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-vinyl-pyridin-3-yl)-propionic acid ethyl ester (0.65 g, to 1.60 mmol) in ethanol (25 ml) at -78°C. Then, after the mixture was barbotirovany O2within 5 minutes, and then N2within 15 minutes. The mixture NaBH4(0,30 g, 8,00 mmol) in water was carefully added to the mixture at -78°and the reaction mixture was left to stand until the temperature reaches 0°C. the Stirring was continued for 3 hours. Acetone was added (10 ml)and the reaction mixture was evaporated to 1/3 of the initial volume. Added 50% NaCl (aqueous), and the mixture was extracted with dichloromethane, dried and concentrated under reduced pressure to obtain crude 3-(6-tert-butoxycarbonyl the amino-5-hydroxymethyl-pyridin-3-yl)-mercaptomethyl-propionic acid ethyl ester (0,38 g, 63%).

and) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-hydroxy-methyl-pyridin-3-yl)-propionic acid ethyl ester

A mixture of crude 3-(6-tert-butoxycarbonylamino-5-hydroxy-methyl-pyridin-3-yl)-mercaptomethyl-propionic acid ethyl ester (0,38 g, 1.0 mmol) and knso3(0.11 g, 1.1 mmol) in acetic anhydride (1 ml) was stirred at room temperature for 5 hours. Then added NH4Cl (saturated) and water. The mixture was extracted with dichloromethane, dried and concentrated under reduced pressure. After flash chromatography (CH2Cl2, 2.5% Meon in CH2Cl2) received a 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-5-hydroxymethyl-pyridin-3-yl)-propionic acid ethyl ester (0,23 g, 60%).

K) 3-(6-Amino-5-hydroxymethyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid

A solution of 3-(6-tert-butoxycarbonylamino-5-hydroxymethyl-pyridin-3-yl)-2-mercaptomethyl-propionic acid ethyl ester (50 mg, 135 mmol) in concentrated HCl (2 ml) was boiled under reflux for 60 minutes. The reaction mixture was cooled and concentrated under reduced pressure to obtain specified in the title compound as Hcl salt (37 mg, 98,3%).

1H NMR (400 MHz, D2O): δ 2.70-2.95 (m, 5H), 4.65 (s, 2H), 7.68 (s, 1H), 7.88 (s, 1H).

MS (+) 244 (M+1).

Example 24

2-Mercaptomethyl-3-pyrrolin the n-3-yl-propionic acid

a) (1-Benzyl-pyrrolidin-3-yl)-methanol

Red-Al (160 ml of a 3.5 M solution in toluene, 560 mmol) was added to a solution of 1-benzyl-5-oxo-pyrrolidin-3-carboxylic acid (20 g, 91 mmol) in dry THF (650 ml) in an argon atmosphere. The reaction mixture is boiled under reflux for 2.5 hours and then poured into a mixture of crushed ice and NaOH (20%). The phases were separated, the aqueous phase was extracted with toluene, and the combined organic extracts were dried and concentrated under reduced pressure to obtain 17.9 g of the crude product as a yellow oil.

b) 3-Hydroxymethyl-pyrrolidin-1-carboxylic acid tert-butyl methyl ether

10%Pd-C (6,1 g) and ammonium formate (10 g, 158 mmol) was added to a solution of (1-benzyl-pyrrolidin-3-yl)-methanol (6,1 g, 32 mmol) in methanol (220 ml) in an argon atmosphere. After boiling under reflux for 15 minutes, the reaction mixture while it is still warm, filtered through a gasket from celite, then celite was washed with methanol, and the combined organic phases were concentrated. The residue was dissolved in THF (35 ml) and water (35 ml), the solution was cooled to 0°and added To2CO3(22 g, 159 mmol) and di-tert-BUTYLCARBAMATE (6,95 g, 32 mmol). The reaction mixture was stirred at room temperature overnight. THF was removed under reduced pressure, was added water, and the aqueous phase was extracted with EtOAc. The combined organic fazzolari and concentrated under reduced pressure to obtain with 4.64 g of the crude product. After flash chromatography (heptane/EtOAc: 1/0→68/32) of the crude product obtained 3-hydroxymethyl-pyrrolidin-1-carboxylic acid tert-butyl ether (3,82 g, 60%) as a colourless oil.

C) 3-Triftoratsetilatsetonom-pyrrolidin-1-carboxylic acid tert-butyl methyl ether

Methanesulfonanilide (0.4 ml, 5.17 mmol) was added dropwise to a solution of 3-hydroxymethyl-pyrrolidin-1-carboxylic acid tert-butyl ester (1 g, equal to 4.97 mmol) and triethylamine (1.04 million ml, 7,46 mmol) in CH2Cl2(15 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight. After filtering was added CH2Cl2and the organic phase is washed with 1 M HCl, dried and concentrated under reduced pressure to obtain 3-triftoratsetilatsetonom-pyrrolidin-1-carboxylic acid tert-butyl ester (1.4 g, 97%).

g) 3-methyl bromide-pyrrolidin-1-carboxylic acid tert-butyl methyl ether

A mixture of 3-triftoratsetilatsetonom-pyrrolidin-1-carboxylic acid tert-butyl ether (3,85 g of 13.8 mmol) and LiBr (3,61 g, 42 mmol) in dry acetone (30 ml) was boiled under reflux overnight. The reaction mixture was left to cool to room temperature, filtered and concentrated. The residue was dissolved in CH2CL2and washed with water, dried and concentrated under reduced pressure to receipt the m 6 g of the crude product. As a result of clearing flash chromatography (heptane/EtOAc: 1/0→68/32) received 3-methyl bromide-pyrrolidin-1-carboxylic acid tert-butyl ether (2,84 g, 78%) as a colourless oil.

d) 2-(1-tert-Butoxycarbonyl-pyrrolidin-3-ylmethyl)-malonic acid diethyl ester

Diethylmalonate (1,93 ml, 12.7 mmol) was added dropwise to a solution of NaH (60%; 0.51 g, 12.8 mmol) in dry THF (15 ml) at 0°C. the mixture was stirred at room temperature for 1 hour, after which it was added to heated under reflux a mixture of 3-methyl bromide-pyrrolidin-1-carboxylic acid tert-butyl ester (2.8 g, 10.6 mmol) in dry THF (30 ml). The reaction mixture was heated under reflux for 19 hours and then concentrated almost to dryness. Was added water (1 l)and the product was extracted with CH2Cl2. The combined organic phase was dried and concentrated under reduced pressure to obtain 3.3 g of the crude product. As a result of clearing flash chromatography (CH2Cl2/EtOAc: 1/0 68/32) received 2-(1-tert-butoxycarbonyl-pyrrolidin-3-ylmethyl)-malonic acid diethyl ester (1.64 g, 45%).

e) 2-(1-tert-Butoxycarbonyl-pyrrolidin-3-ylmethyl)-malonic acid monotropy ether

A solution of KOH (0.26 g, 4.6 mmol) in ethanol (7 ml) was added to a solution of 2-(1-tert-butoxycarbonyl-pyrrolidin-3-ylmethyl)-malonic acid dietrologia (1.52 g, 4.4 mmol) in ethanol (7 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight, concentrated under reduced pressure, and the residue was dissolved in water (500 ml). The aqueous layer was washed with diethyl ether, acidified to pH 3 with 0.5 M HCl and was extracted with diethyl ether. The organic phase was dried and concentrated under reduced pressure to obtain 2-(1-tert-butoxycarbonyl-pyrrolidin-3-ylmethyl)-malonic acid of monoethylene ester (1.13 g, 81%).

g) 3-(2-Etoxycarbonyl-allyl)-pyrrolidine-1-carboxylic acid tert-butyl methyl ether

Diethylamine (0,34 ml, 3.3 mmol) was added to a mixture of 2-(1-tert-butoxy-carbonyl-pyrrolidin-3-ylmethyl)-malonic acid of monoethylene ether (0,69 g, 2,19 mmol) in 36% aqueous solution of formaldehyde (with 0.27 ml, 3.5 mmol), CH2Cl2(1.6 ml) and water (1.6 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight, poured into ice water (500 ml) and was extracted with CH2Cl2. The combined organic phases are washed with 5% NaHCO3, was dried and concentrated under reduced pressure to obtain 3-(2-etoxycarbonyl-allyl)-pyrrolidine-1-carboxylic acid tert-butyl ester (0.55 g, 87%) as a colorless oil.

C) 3-(3-Acetylmethadol-2-etoxycarbonyl-propyl)-pyrrolidine-1-carboxylic acid tert-butyl methyl ether

Teoksessa acid (5 ml), cooling the ing to 0° C, was added to a mixture of 3-(2-etoxycarbonyl-allyl)-pyrrolidine-1-carboxylic acid tert-butyl ether (0,72 g, 2.54 mmol) and triethylamine (0.37 to ml, to 2.67 mmol) at 0°C. the Reaction mixture was stirred at 0°C for 30 minutes, at room temperature for 23 hours and then poured into ice water (400 ml). The aqueous layer was extracted with CH2Cl2. The combined organic phases were washed with saturated NaHCO3, was dried and concentrated under reduced pressure. The crude product was purified flash chromatography (CH2CL2/EtOAc: 1/0 68/32) to obtain 3-(3-acetylmethadol-2-etoxycarbonyl-propyl)-pyrrolidine-1-carboxylic acid tert-butyl ether (0,72 g, 79%) as oil.

and) 2-Mercaptomethyl-3-pyrrolidin-3-yl-propionic acid

A solution of 3-(3-acetylmethadol-2-etoxycarbonyl-propyl)-pyrrolidine-1-carboxylic acid tert-butyl ester (0.52 g, 1,45 mmol) in concentrated HCI (15 ml) was boiled under reflux in an argon atmosphere for 1 hour. The reaction mixture was left to cool to room temperature and concentrated under reduced pressure to get diastereomeric mixture specified in the connection header in the form of cleaners containing hydrochloride salt (0.33 g, 100%).

1H NMR (500 MHz, D2O): δ 1.60-1.92 (m, 3H), 2.19-2.32 (m, 1H), 2.32-2.42 (m, 1H), 2.66-2.83 (m, 3H), 2.84-2.96 (m, 1H), 3.23-3.32 (m, 1H), 3.40-3.58 (m, 2H).

MS (+) 190 (M+1.

Example 25

3-(CIS-4-Amino-cyclopent-2-enyl)-2-mercaptomethyl-propionic acid

a) CIS-Methanesulfonic acid 4-tert-butoxycarbonylamino-cyclopent-2-animationy ether

Methanesulfonanilide (0,76 ml, 9.8 mmol) was added to a solution of CIS-(4-hydroxymethyl-cyclopent-2-enyl)-carbamino acid tert-butyl ester (2 g, 9.4 mmol) and triethylamine (1,96 ml, 14.1 mmol) in CH2Cl2(30 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight. After filtering was added CH2Cl2and the organic phase is washed with 1 M HCl, dried and concentrated under reduced pressure to obtain CIS-methanesulfonic acid 4-tert-butoxycarbonylamino-cyclopent-2-teletrabajo ester (2.64 g, 96%).

b) CIS-(4-methyl bromide-cyclopent-2-enyl)-carbamino acid tert-butyl methyl ether

A mixture of CIS-methanesulfonic acid 4-tert-butoxycarbonylamino-cyclopent-2-teletrabajo ether (of 2.51 g, 8.6 mmol) and LiBr (2.24 g of 25.8 mmol) in dry acetone (20 ml) was boiled under reflux. The reaction mixture was left to cool to room temperature, filtered and concentrated. The residue was dissolved in CH2Cl2and washed with water, dried and concentrated under reduced pressure to obtain CIS-(4-methyl bromide-cyclopent-2-enyl)-carbamino acid tert-butyl ether (2,23 g, 94%).

in) 2(CIS-4-tert-Butoxycarbonylamino-cyclopent-2-animetal)-malonic acid diethyl ester

Diethylmalonate (1,29 ml, 8.5 mmol) was added to a mixture of NaH (60%, 0.34 g; 8.5 mmol) in DMF (10 ml). After stirring at room temperature for 15 minutes was added a solution of CIS-(4-methyl bromide-cyclopent-2-enyl)-carbamino acid tert-butyl ether (1,95 g, 7.1 mmol) in DMF (12 ml)and the reaction mixture was stirred at 60°C for 19 hours. Added EtOAc, and the organic phase was extracted with water and brine, dried and concentrated under reduced pressure to get 2,44 g of the crude product. As a result of clearing flash chromatography (CH2Cl2/EtOAc: 1/0 68/32) received 2-(4-CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-malonic acid diethyl ester (1.47 g, 58%).

g) 2-(CIS-4-tert-Butoxycarbonylamino-cyclopent-2-animetal)-malonic acid monotropy ether

A solution of KOH (0,19 g; 3.4 mmol) in ethanol (6 ml) was added to a solution of 2-(2-CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-malonic acid diethyl ester (1,15 g; 3.2 mmol) in ethanol (6 ml) at 0°C. the Reaction mixture was stirred at room temperature over night, concentrated and added to a mixture of ice water (400 ml). The aqueous phase is washed with diethyl ether (emulsion formed during the extraction, the treated brine to achieve good separation of phases), acidified to pH 3 with 0.5 M HCl and was extracted with diethyl ether. About the organic phase was dried and concentrated under reduced pressure to obtain 2-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-malonic acid of monoethylene ether (0,86 g, 81%) as white crystals.

d) 2-(CIS-4-tert-Butoxycarbonylamino-cyclopent-2-animetal)-acrylic acid ethyl ester

Diethylamine (0,31 ml, 3.0 mmol) was added to a mixture of 2-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-malonic acid of monoethylene ester (0.66 g, 2.0 mmol) in 36% aqueous solution of formaldehyde (0.25 ml, 3.2 mmol), CH2Cl2(1.6 ml) and water (1.6 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight, poured into ice water (40 ml) and was extracted with CH2Cl2. During the extraction, phase separation was improved by the addition of brine. The combined organic extracts washed with 5% NaHCO3, was dried and concentrated under reduced pressure to obtain 2-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-acrylic acid ethyl ester (0.56 g, 94%) as oil.

e) 2-Ametisoidukitel-3-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-enyl)-propionic acid ethyl ester

Teoksessa acid (4 ml), cooled to 0°S, was added to a mixture of 2-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-animetal)-acrylic acid ethyl ester (0.56 g, 1.9 mmol) and triethylamine (of 0.28 ml, 2.00 mmol) at 0°C. the Reaction mixture was stirred at 0°C for 30 minutes at room temperature for 19 hours and then poured into ice water (400 ml). Water SL the St was extracted with CH 2Cl2. The organic phase was washed with saturated NaHCO3, was dried and concentrated under reduced pressure to obtain 1.7 g of the crude product. As a result of clearing flash chromatography (CH2Cl2/EtOAc: 1/0 68/32) received 2-acetylsalicylate-3-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-enyl)-propionic acid ethyl ester (0,46 g, 65%).

g) 3-(CIS-4-Amino-cyclopent-2-enyl)-2-mercaptomethyl-propionic acid

A solution of 2-acetylsalicylate-3-(CIS-4-tert-butoxycarbonylamino-cyclopent-2-enyl)-propionic acid ethyl ester (86 mg, 0.23 mmol) in concentrated HCl (5 ml) was boiled under reflux in an argon atmosphere for 1 hour. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to get diastereomeric mixture specified in the connection header in the form of cleaners containing hydrochloride salt (65 mg).

1H NMR (400 MHz, D2O): δ 1.35-1.47 (m, 2H), 1.60-1.72 (m, 1H), 1.74-1.92 (m, 2H), 1.92-2.04 (m, 1H), 2.67-2.92 (m, 10H), 4.34-4.43 (m, 2H), 5.79-5.85 (br s, 2H), 6.12 (d, 1H), 6.18 (d, 1H).

Example 26

2-Mercaptomethyl-3-piperazine-1-yl-propionic acid

a) 4-(2-Etoxycarbonyl-allyl)-piperazine-1-carboxylic acid tert-butyl methyl ether

To a solution of 2-methyl bromide-acrylic acid ethyl ester (of 1.93 g, 10 mmol) in N,N-dimethylformamide (25 ml) was added piperazine-1-carboxylic acid tert-butile the initial ester (1.86 g, 10 mmol)and then dropwise added ethyldiethanolamine (1,71 ml, 10 mmol). After stirring for 16 hours at room temperature the solvent was removed under reduced pressure and added water (50 ml) and dichloromethane (50 ml). After stirring for 2 minutes, the layers were separated, the organic layer was washed with water and brine and dried over magnesium sulfate. The solvent was removed under reduced pressure to obtain crude 4-(2-etoxycarbonyl-allyl)-piperazine-1-carboxylic acid tert-butyl ether (2,456 g, 82%).

b) 4-(3-Acetylmethadol-2-etoxycarbonyl-propyl)-piperazine-1-carboxylic acid tert-butyl methyl ether

To the crude 4-(2-etoxycarbonyl-allyl)-piperazine-1-carboxylic acid tert-butyl ether (2,45 g, 8.2 mmol) was added toluxury acid (7.5 ml) in an argon atmosphere. The mixture was cooled to 0°and was added dropwise a triethylamine (1,14 ml, 8.2 mmol). The mixture was then stirred at room temperature for 2 days and added toluxury acid (2.5 ml). Stirring was continued for 1 day, then carefully added aqueous saturated solution of sodium bicarbonate until neutral, and the mixture was extracted three times with dichloromethane. The combined organic layers were washed twice aqueous saturated sodium bicarbonate and once with brine. After drying over sulfate mage the Oia, the solvent was removed under reduced pressure, and the residue was purified column chromatography (silica gel, 1.7% methanol in dichloromethane) to give 4-(3-acetylmethadol-2-etoxycarbonyl-propyl)-piperazine-1-carboxylic acid tert-butyl ester (0.29 grams, 9,4%).

a) 2-Mercaptomethyl-3-piperazine-1-yl-propionic acid

To 4-(3-acetylmethadol-2-etoxycarbonyl-propyl)-piperazine-1-carboxylic acid tert-butyl ether (0,095 g, 0.25 mmol) was added saturated with argon hydrochloric acid (3 ml, 37%), and the mixture was heated in an argon atmosphere to a temperature of reflux distilled for 2.5 hours. The solution was concentrated under reduced pressure to obtain specified in the connection header in the form dihydrochloride salt (0,071 g, quantitatively).

1H NMR (400 MHz, D2O): δ 3.86-3.49 (m, 10H), 3.36-3.28 (m, 1H), 3.01-2.88 (m, 2H).

MS 205 (M+H).

Example 27

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-pentane acid

a) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid diethyl ester

To a stirred suspension of copper iodide (5,71 g, 30 mmol) in diethyl ether (120 ml) in an argon atmosphere was added ethylmagnesium (3 M solution in diethyl ether, 20 ml, 20 mmol) over 10 minutes at 0°C. After 5 minutes of stirring the mixture was cooled to -78°and was added dropwise a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester (vs. 5.47 g, 15.0 m is ol) in tetrahydrofuran (60 ml) for 0.5 hours. During this period of time was added tetrahydrofuran (40 ml). The mixture was left to mix at -40°C for 2.5 hours. Then added with vigorous stirring a solution of ammonium chloride (5%) in aqueous ammonia solution (5%), allowing access of air, and the mixture was heated to room temperature. The mixture was extracted twice with ethyl acetate. The combined organic layers were washed sequentially with an aqueous solution of ammonia (5%) and brine and was dried over magnesium sulfate. The solvent was removed under reduced pressure, and the residue was recrystallized from a mixture of diisopropyl ether and tetrahydrofuran (5:1, vol/about.) obtaining 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid diethyl ester (3,23 g, 55%).

b) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid monotropy ether

2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid diethyl ester (3,23 g, 8,19 mmol) was dissolved in a mixture of dichloromethane (12 ml) and ethanol (24 ml, 99.5%pure). To this solution was added dropwise a solution of potassium hydroxide (0,528 g, 87%, 8.20 mmol) in ethanol (16 ml, 99.5%pure) at 0°for 0.5 hour. Stirring was continued for 16 hours, during which time the mixture was left to warm to room temperature. After concentration under reduced pressure to 5-10 ml of relax is whether water (50 ml), and the resulting emulsion was stirred for 0.5 hour and then filtered. The filtrate was washed twice with a mixture of ethyl acetate and diethyl ether (2:1, vol/vol.). The aqueous layer was acidified by adding citric acid solution in water (10%) to achieve a pH of from 4 to 5 and was extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, and the solvent was removed under reduced pressure to obtain 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid of monoethylene ether (2,47 g, 82%).

a) 2-[1-(6-tert-Butoxycarbonyl-pyridine-3-yl)-propyl]-acrylic acid ethyl ester.

To a solution of 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-propyl]-malonic acid of monoethylene ether (2,45 g, 6,69 mmol) in dichloromethane (11 ml) was added an aqueous solution of formaldehyde (0,49 ml, 37%) and diethylamine (of 0.62 ml, 6.4 mmol) at 0°C. After vigorous stirring the mixture for 16 hours was added water (40 ml) and ethyl acetate (40 ml). After an additional 2 minutes of mixing at room temperature the layers were separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with saturated aqueous sodium hydrogen carbonate solution and brine. After drying over magnesium sulfate the solvent was removed under reduced pressure to obtain 2-[1-(6-tert-butoxycarbonyl-is iridin-3-yl)-propyl]-acrylic acid ethyl ester (1,21 g, 54%).

g) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester

2-[1-(6-tert-Butoxycarbonyl-pyridine-3-yl)-propyl]-acrylic acid ethyl ester (1.20 g, 3,59 mmol) was dissolved in thioglucose acid (4 ml) in an argon atmosphere was added triethylamine (of 0.56 ml, 4.0 mmol). The mixture was heated to 60°C. After 22 hours added toluxury acid (2 ml). After 14 hours the mixture was cooled to room temperature and was slowly added saturated aqueous sodium bicarbonate to obtain a neutral solution. It was extracted three times with ethyl acetate. The combined organic layers washed with aqueous saturated sodium bicarbonate and brine, and dried over magnesium sulfate. After concentration under reduced pressure the residue was purified column chromatography (silica gel, 1.7% methanol in dichloromethane) to give 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester (1.22 g, 83%).

The mixture of diastereomers was separated preparative chiral chromatography, using a column CHIRALPAK AD (250×4.6 mm) as the stationary phase and hexane (mixture of isomers)/ethanol (85:15)containing diethylamine (0,05%) at a flow rate of 1 ml/min with a concentration of a sample of 5 mg/ml

The enantiomeric excess was determined by analytical chiral VE the X on the same column using a mixture of hexane/ethanol (85:15) as eluent.

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester:

Retention time of 10.72 minutes, it (enantiomeric excess) = 99%.

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/B:

Retention time 14,41 minutes, it = 98%.

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/In:

Retention time 23,01 minutes, it = 98%.

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/G:

Retention time 29,83 minutes, it = 97%.

d) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-pentane acid

Hydrochloric acid (38%, 4 ml) was added to 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ether level (0.041 g, 0.1 mmol) in an argon atmosphere, and the mixture was heated under reflux for 4 hours. After concentration under reduced pressure and drying (45°C, 0.3 mbar (0,3×10-8PA)) were specified in the title compound in the form of cleaners containing hydrochloride salt (0,027 mg, 97%).

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-pentane acid/A.

This compound was obtained from 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/And as described to enter the.

1H NMR (300 MHz, D2O): δ 7.79 (dd, 1H), 7.69 (d, 1H), 7.05 (d, 1H), 2.81 (m, 2H), 2.51 (m, 2H), 1.66 (m, 2H), 0.72 (t, 3H).

MS + 241 (M+H).

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-pentane acid/B

This compound was obtained from 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/B in the manner described above.

1H NMR (300 MHz, D2O): δ 7.85 (dd, 1H), 7.64 (d, 1H), 7.01 (d, 1H), 2.98-2.64 (m, 4H), 1.89 (m, 1H), 1.60 (m, 1H), 0.74 (t, 3H).

MS + 241 (M+H).

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-pentane acid/

This compound was obtained from 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/In the same manner as described above.

1H NMR (300 MHz, D2O): δ 7.85 (dd, 1H), 7.64 (d, 1H), 7.01 (d, 1H), 2.98-2.64 (m, 4H), 1.89 (m, 1H), 1.60 (m, 1H), 0.74 (t, 3H).

MS + 241 (M+H).

3-(6-Amino-pyridin-3-yl]-2-mercaptomethyl-pentane acid/G

This compound was obtained from 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-pentanol acid ethyl ester/G in the manner described above.

1H NMR (300 MHz, D2O): δ 7.79 (dd, 1H), 7.69 (d, 1H), 7.05 (d, 1H), 2.81 (m, 2H), 2.51 (m, 2H), 1.66 (m, 2H), 0.72 (t, 3H).

MS + 241 (M+H).

Example 28

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-4-methyl-pentane acid

a) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid diethyl ester

To a mixed solution of copper cyanide (3.58 g, 40 mmol) in tetrahydrofuran (50 ml) at -15°solution was added isopropylacrylamide (40 ml, 2 M, 80 mmol) in tetrahydrofuran in an argon atmosphere for 15 minutes. After stirring for a further 15 minutes, was added dropwise a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester (3.78 g, 10.4 mmol) in tetrahydrofuran (50 ml) for 15 minutes. After stirring for 16 hours the mixture was left to warm to room temperature. Then with vigorous stirring solution was added ammonium chloride (5%) in aqueous ammonia solution (5%), allowing access of air. The mixture was extracted twice with ethyl acetate. The combined organic layers were washed sequentially with an aqueous solution of ammonia (5%) and brine and was dried over magnesium sulfate. The solvent was removed under reduced pressure, and the residue was purified column chromatography (silica gel, 1.7% methanol in dichloromethane) to give the crude 2-[1-(tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid diethyl ester (2.85 g, 25%).

b) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid monotropy ether

To a solution of 2-[1-(tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid diethyl ester (1,569 g of 3.84 mm is l) in a mixture of dichloromethane (6 ml) and ethanol (12 ml, 95%) at 0°C was added dropwise a solution of potassium hydroxide (0,272 g, 85%, 4.2 mmol) in ethanol (6 ml, 95%) for 40 minutes. After stirring for a further 1 hour the mixture was left to warm to room temperature and stirring continued for 18 hours. Then added water (30 ml) and dichloromethane (30 ml), and after 3 minutes of stirring, the layers were separated. The organic layer was extracted once with water and the combined aqueous layer was washed once with ether. Then added an aqueous solution of citric acid to bring the pH to 4, and the solution was extracted three times with ethyl acetate. The combined organic layers were washed with brine and dried over magnesium sulfate. After evaporation of the solvent obtained crude 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid monotropy ether (0,94 g, 64%).

a) 2-[1-(6-tert-Butoxycarbonyl-pyridine-3-yl)-2-methyl-propyl]-acrylic acid ethyl ester

To a solution of 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-methyl-propyl]-malonic acid of monoethylene ether (0,94 g, 2,47 mmol) in dichloromethane (4 ml) was added an aqueous solution of formaldehyde (of 0.18 ml, 37%) and diethylamine (0,23 ml, 2.32 mmol) at 0°C. After vigorous stirring the mixture for 16 hours was added water (30 ml) and ethyl acetate (30 ml). After additional stirring for 2 minutes at room tempera is ur layers were separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with saturated aqueous sodium hydrogen carbonate solution and brine. After drying over sodium sulfate the solvent was removed under reduced pressure, and the residue was purified column chromatography (silica gel, 1.7% methanol in dichloromethane) to give 2-[1-(6-tert-butoxycarbonyl-pyridine-3-yl)-2-methyl-propyl]-acrylic acid ethyl ester (0.24 g, 28%).

g) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-methyl-pentanol acid ethyl ester

2-[1-(6-tert-Butoxycarbonyl-pyridine-3-yl)-2-methyl-propyl]-acrylic acid ethyl ester (0,431 g of 1.24 mmol) was dissolved in thioglucose acid (4 ml) in an argon atmosphere was added triethylamine (of 0.21 ml, 1.5 mmol). The mixture was heated to 60°C. After 24 hours was added toluxury acid (2 ml). After 20 hours the mixture was left to cool to room temperature and was slowly added to a saturated solution of sodium bicarbonate to obtain a neutral solution. It was extracted three times with ethyl acetate. The combined organic layers washed with aqueous saturated sodium hydrogen carbonate solution and brine and dried over magnesium sulfate. After concentration under reduced pressure the residue was purified column chromatography (silica gel, 1.7% methanol in dichloromethane)to give 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-methyl-pentanol acid ethyl ester (0,284 g, 54%) as a mixture of diastereoisomers (6:1).

d) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-4-methyl-pentane acid

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-methyl-pentanol acid ethyl ester (of 0.085 g, 0.20 mmol) was dissolved in saturated argon hydrochloric acid (5 ml, 37%) and heated under reflux in an argon atmosphere for 5.5 hours. After concentration under reduced pressure and drying at 45°/0.3 mbar (0,3×10-8PA) has been specified in the title compound as a mixture of diastereoisomers (6:1) in the form of cleaners containing hydrochloride salt (0,058 g, 99%).

1H NMR (300 MHz, D2O): δ 7.81 (m, 1H), 7.62 (m, 1H), 6.98 (m, 1H), 3.18-3.06 (m, 1H), 2.93-2.40 (m, 3H), 2.23-1.93 (m, 1H), 0.93-0.74 (m, 6H).

MS 255 (M+H).

Example 29

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-3-phenyl-propionic acid

a) 2-[(6-tert-Butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-malonic acid diethyl ester

To intensively stirred suspension of copper cyanide (I) (1,71 g, 19,06 mmol) in dry THF (18 ml) solution was added phenylmagnesium (12,7 ml of 3 M in ether, 38,11 mmol) at 0°C in argon atmosphere. The mixture was left to warm to room temperature, it was formed a dark brown solution. After 150 minutes was added at 0°With a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester (1,74 g, 4.76 mmol) and the dry THF (19 ml). The mixture was left to mix for 3 days, then was added aqueous ammonium chloride. The aqueous layer was separated and was extracted with ethyl acetate. The combined organic layers were washed with brine and dried. After removal of solvents in vacuo received the remainder, which is suspended in hexane. After filtering off the crystals obtained 2-[(6-tert-butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-malonic acid diethyl ester (1.85 g, 88%).

b) 2-[(6-tert-Butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-malonic acid monotropy ether

A solution of KOH (0,226 g, 4,11 mmol) in ethanol (14 ml) was added to a solution of 2-[(6-tert-butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-malonic acid diethyl ester (1,82 g, 4,11 mmol) in ethanol (12 ml) and methylene chloride (13 ml) at 0°C. the Mixture was stirred over night at room temperature. Another CON (80 mg, dissolved in 3 ml ethanol) was added at 0°C. the Reaction mixture was stirred for another 18 hours. The mixture was concentrated under reduced pressure and to the residue was added ethyl acetate and 0.5 M HCl. The organic layer was washed with brine and dried. After filtration and evaporation in vacuo the residue is suspended in hexane. After filtering off the crystals obtained 2-[(6-tert-butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-malonic acid monotropy ether (1.7 g, 98%).

a) 2-[(6-the pet-Butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-acrylic acid ethyl ester

Diethylamine (0.52 g, 5,04 mmol) was added to a mixture of 2-[(6-tert-butoxycarbonyl-amino-pyridin-3-yl)-phenyl]-malonic acid of monoethylene ether (1.7 g, 4.1 mmol) and 37% aqueous formaldehyde (0,42 g, 5.6 mmol) in methylene chloride (6.5 ml) at 0°C. the Mixture was stirred over night at room temperature and then was diluted with ethyl acetate. The organic layer is washed with aqueous saturated sodium bicarbonate and brine and dried. After filtration and evaporation in vacuo the crude product was purified flash chromatography (CH2Cl2) to obtain 2-[(6-tert-butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-acrylic acid ethyl ester (0.36 g, 23%).

g) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-3-phenyl-propionic acid ethyl ester

The triethylamine (0,105 g, 1.04 mmol) was added to a solution of 2-[(6-tert-butoxycarbonylamino-pyridine-3-yl)-phenyl-methyl]-acrylic acid ethyl ester (0.36 g, were 0.94 mmol) in teoksessa acid (4 ml) at 0°C. the Mixture was heated at 45°C for 24 hours. Added ethyl acetate, and the organic phase is washed with aqueous saturated sodium bicarbonate and brine and dried. After filtration and evaporation in vacuo the crude product was purified flash chromatography (toluene/ethyl acetate, 1:05:1) to give pure 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-Piri is INF-3-yl)-3-phenyl-propionic acid ethyl ester (0,314 g, 73%).

d) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-3-phenyl-propionic acid

2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-3-phenyl-propionic acid ethyl ester (58 mg, 0.125 mmol) was dissolved in concentrated HCl (3.0 ml). The solution was heated under reflux for 130 minutes in an argon atmosphere. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (41 mg, 100%).

1H NMR (500 MHz, D2O): δ 8.03-7.83 (m, 2H), 7.5-7.3 (m, 5H), 7.05-6.95 (m, 1H), 4.27-4.15 (m, 1H), 3.6-3.45 (m, 1H), 2.84-2.58 (m, 2H).

MS (+) 289 (M+1).

Example 30

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-3-phenyl-butyric acid

a) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-malonic acid diethyl ester

To intensively stirred suspension of copper cyanide (I) (0.66 g, to 7.32 mmol) in dry THF (5 ml) solution was added benzylacrylamide (5 ml, 2,93 M in ether, 14,64 mmol) at 0°C in argon atmosphere. The mixture was left to warm to room temperature, it was formed a dark brown solution. After 60 minutes was added at 0°With a solution of 2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethylene)-malonic acid diethyl ester (0,67 g and 1.83 mmol) in dry THF (4 ml). The mixture was stirred over night at room temperature, then added an aqueous ammonium chloride. Vodnykh was separated and was extracted with ethyl acetate. The combined organic layers were washed with brine and dried. After removal of solvent in vacuo received a residue, which was purified flash chromatography (CH2CL2/ethyl acetate, 1:0→100:15) to give 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-malonic acid diethyl ester (0,44 g, 53%).

b) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-malonic acid monotropy ether

A solution of KOH (0,067 g of 1.03 mmol) in ethanol (3 ml) was added to a solution of 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-malonic acid diethyl ester (0,44 g, 0.96 mmol) in ethanol (3 ml) and methylene chloride (2 ml) at 0°C. the Mixture was stirred for 48 hours at room temperature. The mixture was concentrated under reduced pressure and to the residue was added ethyl acetate and 0.5 M HCl. The organic layer was washed with brine and dried. As a result of filtration and evaporation in vacuum is obtained 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-malonic acid monotropy ether (0,37 g, 90%).

a) 2-[1-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-acrylic acid ethyl ester

Diethylamine (0,27 ml, at 2.59 mmol) was added to a mixture of 2-[1-(6-tert-butoxycarbonyl-amino-pyridin-3-yl)-2-phenyl-ethyl]-malonic acid of monoethylene ether (0,37 g, 0.86 mmol) and 37% aqueous formaldehyde (0,22 ml, 2.9 mmol) veterinariae (3.5 ml) at 0° C. the Mixture was stirred over night at room temperature. Was added formaldehyde (0,24 ml) and diethylamine (0,24 ml). Stirring at room temperature was continued for 18 hours and then added ethyl acetate and water. The organic layer is washed with aqueous saturated sodium bicarbonate and brine, dried and concentrated under reduced pressure to obtain 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-acrylic acid ethyl ester (0.34 g, 99%).

g) 2-Acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-phenyl-butyric acid ethyl ester

The triethylamine (0,096 g, 0.95 mmol) was added to a solution of 2-[1-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-phenyl-ethyl]-acrylic acid ethyl ester (0.34 g, 0.86 mmol) in teoksessa acid (3.5 ml) at 0°C in argon atmosphere. The mixture was heated at 45°C for 24 hours. Added ethyl acetate, and the organic phase is washed with aqueous saturated sodium bicarbonate and brine and dried. After filtration and evaporation in vacuo the crude product was purified flash chromatography (CH2CL2/ethyl acetate, 1:0 100:5) to give 2-acetylsalicylate-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-phenyl-butyric acid ethyl ester (0,264 g, 65%).

d) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-4-phenyl-butyric acid

2-Acetylanthranilic the l-3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-4-phenyl-butyric acid ethyl ester (0.14 g, 0.3 mmol) was dissolved in concentrated HCl (5.0 ml). The solution was heated under reflux for 4.5 hours in an argon atmosphere. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (100 mg, 98%).

1H NMR (500 MHz, D2O): δ 7.90-7.83 (m, 1H), 7.43 (d, 1H), 7.34-7.18 (m, 3H), 7.17-7.06 (m, 2H), 7.01-6.9 (m, 1H), 3.4-2.5 (m, 6H).

MS (+) 303 (M+1).

Example 31

2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-5-phenyl-pentane acid

a) Ethyl(E,Z)-2-({6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}methyl)-5-phenyl-2-pentenoate

To a suspension of NaH (310 mg, 7,12 mmol, 55% in mineral oil) in THF (25 ml) at 0°C in an atmosphere of argon was added a solution (ethyl-3-{6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}-2-(diethoxyphosphoryl)propanoate (2.55 g, 5,95 mmol) in THF (25 ml). After 1 hour, was added dropwise a solution of 3-phenylpropanal (1,59 g, to 11.9 mmol). The reaction mixture was stirred for 17 hours at room temperature, then extinguished NH4Cl (50 ml, saturated aqueous). The mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried (Na2SO4), filtered and concentrated under reduced pressure. After column chromatography (CH2Cl2/EtOAc 20:1 to 10:1) was obtained ethyl(E,Z)-2-({6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}methyl)-5-phenyl-2-pentenoate (2,49 g, 100%).

b) Ethyl-3-(acetylsalicyl)-2-({6-[(is pet-butoxycarbonyl)amino]-3-pyridinyl}methyl)5-phenyl-2-pentenoate

The triethylamine (1,22 ml, 0,617 mmol) was added to a solution of ethyl-(E,Z)-2-({6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}methyl)-5-phenyl-2-pentenoate (400 mg, 0,597 mmol) in teoksessa acid (10 ml) at 40°C. After stirring for 90 hours the mixture was concentrated under reduced pressure. After column chromatography (CH2Cl2/EtOAc 20:1 to 10:1), then (toluene/EtOAc, 10:1), and then (heptane/EtOAc 2:1) was obtained ethyl-3-(acetylsalicyl)-2-({6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}methyl)-5-phenylpentane (126 mg, 27%) as a mixture of diastereoisomers 1:1.

a) 2-(6-Amino-pyridine-3-ylmethyl)-3-mercapto-5-phenyl-pentane acid

Ethyl-3-(acetylsalicyl)-2-({6-[(tert-butoxycarbonyl)amino]-3-pyridinyl}methyl)-5-phenylpentane (9 mg, 18.5 μmol) was dissolved in concentrated HCl (1 ml) in an argon atmosphere. The solution was heated under reflux for 4.5 hours. After concentration under reduced pressure has been specified in the title compound in the form of cleaners containing hydrochloride salt (6.4 mg, 98%) as a mixture of diastereoisomers 1:1.

1H NMR (400 MHz, D2O): δ 1.78-2.20 (m, 2H), 2.70-3.04 (m, 6H), 6.88, 6.92 (2d, 1H), 7.22-7.39 (m, 5H), 7.52, 7.54 (2d, 1H), 7.69, 7.75 (2d, 1H).

MS (+) 317 (M+1).

Example 32

3-[3-(6-Amino-pyridin-3-yl)-2-etoxycarbonyl-propylsulfonyl]-2-(6-amino-pyridine-3-ylmethyl)-propionic acid ethyl ester

a) 3-[3-(6-tert-Butoxycarbonylamino-pyridine-3-yl)-2-ethoxycarbonyl the l-propylsulfonyl]-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-propionic acid ethyl ester

3-Acetylmethadol-2-(6-tert-butoxycarbonylamino-pyridine-3-yl-methyl)-propionic acid ethyl ester (150 mg, 0,392 mmol) was dissolved in ethanol (15 ml), saturated NH3(gas). After stirring for 160 minutes, the mixture was concentrated under reduced pressure. The residue was dissolved in EtOH (10 ml), then was added a solution of I2in EtOH (0.5 M, 0,784 ml). The reaction mixture was stirred for 30 minutes at room temperature, then diluted with CH2Cl2, washed with saturated PA2S2O3and saturated aqueous Panso3, dried, filtered and concentrated under reduced pressure to obtain 3-[3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-etoxycarbonyl-propylsulfonyl]-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-propionic acid ethyl ester (130 mg, 98%).

b) 3-[3-(6-Amino-pyridin-3-yl)-2-etoxycarbonyl-propylsulfonyl]-2-(6-amino-pyridine-3-ylmethyl)-propionic acid ethyl ester

Ethyl acetate saturated with HCl (gas) (15 ml)was added to a solution of 3-[3-(6-tert-butoxycarbonylamino-pyridine-3-yl)-2-etoxycarbonyl-propyl-disulfonic]-2-(6-tert-butoxycarbonylamino-pyridine-3-ylmethyl)-propionic acid ethyl ester (130 mg, 0,191 mmol) in ethyl acetate (7 ml) at 0°C. the Reaction mixture was left to stand to reach room temperature and was stirred for 19 hours, then pack rivali under reduced pressure. The residue was dissolved in water, was added saturated aqueous PA2CO3and the aqueous phase was extracted with CH2Cl2. The organic layer was dried and concentrated under reduced pressure to obtain specified in the title compound (90 mg, 98%).

1H NMR (400 MHz, CDCl3): δ 1.19 (t, 3H), 1.20 (t, 3H), 2.68-2.86 (m, 6H), 2.90-3.01 (m, 4H), 4.07-4.14 (m, 4H), 6.45 (d, 2H), 7.28 (m, 2H), 7.87 (s, 2H).

MS (+) 479 (M+1).

Example 33

3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-hexanoic acid

a) N-(5-Butyryl-pyridine-2-yl)-2,2-dimethyl-propionamide

A solution of N-(5-bromopyridin-2-yl)-2,2-dimethyl-propionamide (3,582 g, a 13.9 mmol) in diethyl ether (36 ml) was cooled to -78°C in an atmosphere of argon was added dropwise n-utility (1.6 M, 19 ml, 30.4 mmol) in 20 minutes. After 5 minutes after the addition the mixture was left to warm to 0°C. was Added N-methoxy-N-methylbutyrate (3,65 g, 27.8 mmol) in less than 5 minutes, and stirring was continued for 45 minutes. The clear solution was acidified using 2 N. hydrochloric acid to pH 2. After intensive stirring for 15 minutes, the mixture was neutralized with sodium bicarbonate, and the layers were separated. The aqueous layer was extracted three times with diethyl ether. The combined organic layers were washed with brine, dried and concentrated under reduced pressure. The resulting oil was dried under vacuum, generated oil is m pump to remove excess reagent. After flash chromatography (dichloromethane/methanol 60:1) obtained crude N-(5-butyryl-pyridine-2-yl)-2,2-dimethyl-propionamide (2,47 g, 71%) as a pale orange oil, which is not further purified.

b) N-[5-(1-Hydroxy-butyl)-pyridine-2-yl]-2,2-dimethyl-propionamide

To a solution of N-(5-butyryl-pyridine-2-yl)-2,2-dimethyl-propionamide (2,47 g, for 9.95 mmol) in ethanol (20 ml) was added sodium borohydride (185 mg, 5.0 mmol). After 40 minutes of stirring at room temperature the mixture was acidified using 1 N. hydrochloric acid to pH 2 and was stirred for 10 minutes. After neutralization with sodium bicarbonate, the mixture was extracted three times with dichloromethane. The combined organic extracts were washed twice with sodium hydrogen carbonate solution and once with brine. After drying and concentration under reduced pressure the residue was purified flash chromatography (dichloromethane/methanol 30:10 obtaining N-[5-(1-hydroxy-butyl)-pyridine-2-yl]-2,2-dimethyl-propionamide (1,025 g, 41%).

a) 2-{1-[6-(2,2-Dimethyl-propionamide)-pyridine-3-yl]-butyl}-malonic acid monotropy ether

To a solution of N-[5-(1-hydroxy-butyl)-pyridine-2-yl]-2,2-dimethyl-propionamide (1,025 g, 4.09 to mmol) in trichloromethane (12 ml) was added thionyl chloride (6 ml), and the mixture was heated to 55°C for 1 hour. Then the solution was concentrated under reduced pressure to get neocide the aqueous N-[5-(1-chloro-butyl)-pyridine-2-yl]-2,2-dimethyl-propionamide hydrochloride as a colourless solid.

To a solution of diethylmalonate (1.31 g, 8.2 mmol) in dimethylformamide (20 ml) was added sodium hydride (60%dispersion in mineral oil, 350 mg, 8,7 mmol)and the mixture was stirred at room temperature for 15 minutes. After cooling to 0°C was added a solution of intermediate N-[5-(1-chloro-butyl)-pyridine-2-yl]-2,2-dimethyl-propionamide hydrochloride in dimethylformamide (5 ml). After 1 hour of stirring the solution was concentrated under reduced pressure and added ethyl acetate (25 ml), and then a solution of ammonium chloride (Polynesians) for neutralization. The layers were separated, the aqueous layer was extracted with ethyl acetate, and the combined organic layers were washed three times with water and brine and dried. After concentration under reduced pressure the residue was filtered through silica gel using a mixture of dichloromethane/methanol (30:1) as solvent.

This crude 2-{1 -[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-butyl}-malonic acid diethyl ester was dissolved in a mixture of dichloromethane (2.5 ml) and ethanol (5 ml) at 0°and the solution was added potassium hydroxide (87%, 111 mg, 5.4 mmol) in ethanol (4 ml). This mixture was left to slowly warm to room temperature, and stirring was continued for 24 hours. Then added dichloromethane (20 ml), water (20 ml) and brine (3 ml). The mixture was intensively stirred for 2 minutes. Satisloh shared, the aqueous layer was washed with dichloromethane. The combined aqueous layers were acidified (pH 5) citric acid and was extracted three times with dichloromethane. The combined organic layers were washed with brine and dried. After concentration under reduced pressure, the obtained crude 2-{1-[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-butyl}-malonic acid monotropy ester (258 mg, 41%).

g) 2-{1-[6-(2,2-Dimethyl-propionamide)-pyridine-3-yl]-butyl}-acrylic acid ethyl ester

To a solution of 2-{1 -[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-butyl}-malonic acid of monoethylene ether (712 mg, of 1.95 mmol) in THF (6.5 ml) at 0°With added formaldehyde (37% in water, 0.3 ml) for 5 minutes. Stirring was continued for 10 min, then was added dropwise piperidine (of 0.26 ml, 2,63 mmol) in 10 minutes. The mixture was left to warm overnight to room temperature. After 14 hours the mixture was concentrated under reduced pressure to one-third of its volume, and then was added ether and water (15 ml each). After 2 minutes of intensive mixing, the layers were separated, the aqueous layer was extracted with ether, and the combined organic layers were washed with water, 4%citric acid, water and brine. After drying and concentration under reduced pressure was obtained 2-{1-[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-butyl}-acrylic acid ethyl ester (550 mg, 85%) as a demon who Vatan solids.

d) 2-Acetylsalicylate-3-[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-hexanoic acid ethyl ester

2-{1-[6-(2,2-Dimethyl-propionamide)-pyridine-3-yl]-butyl}-acrylic acid ethyl ester (550 mg, of 1.65 mmol) was dissolved in argon atmosphere in thioglucose acid (4 ml)and dropwise added triethylamine (of 0.24 ml, 1.7 mmol). After stirred for 16 hours at 50°With added toluxury acid (2 ml), and stirring was continued for 14 hours. The solution was cooled to room temperature and neutralized by adding sodium hydrogen carbonate solution. The mixture was extracted three times with ethyl acetate, the combined extracts were washed with sodium hydrogen carbonate solution and brine. After drying and concentration under reduced pressure the residue was purified flash chromatography (dichloromethane/methanol, 60:1) to obtain 2-acetylsalicylate-3-[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-hexanoic acid ethyl ester (619 mg, 92%) as a viscous oil.

e) 3-(6-Amino-pyridin-3-yl)-2-mercaptomethyl-hexanoic acid

2-Acetylsalicylate-3-[6-(2,2-dimethyl-propionamide)-pyridine-3-yl]-hexanoic acid ethyl ester (40,4 mg, 99 μmol) was dissolved in argon atmosphere in aqueous hydrochloric acid (37%, to 4.2 ml) and was heated under reflux for 2 hours. After concentration under reduced pressure of 0.3 Torr, 40°Prov.) has been specified in the title compound in the form of cleaners containing hydrochloride salt (28 mg, 97%).

1H NMR (300 MHz, D2O): δ 7.86 (d, 1H), 7.65 (d, 1H), 7.02 (t, 1H), 2.98-2.42 (m, 4H), 1.85-1.54 (m, 2H), 1.19-1.00 (m, 2H), 0.87-0.77 (m, 3H).

MS (+) 225 (M+1).

Example 34

3-(2-Amino-thiazol-5-yl)-2-mercaptomethyl-propionic acid

a) 2-(2-Amino-thiazole-5-ylmethylene)-malonic acid diethyl ester

To a solution of 2-amino-thiazole-5-carbaldehyde (47%; 6 g; 23 mmol) in CH2Cl2(30 ml) and DMF (30 ml) was added over molecular sieve 4diethylmalonate (3.5 ml; 23 mmol), piperidine (1.1 ml, 11.5 mmol) and acetic acid (0.7 ml, 11.5 mmol). The reaction mixture was stirred at room temperature for 96 hours. Then added EtOAc and the reaction mixture was filtered through celite to remove the precipitate formed. To the filtrate was added EtOAc (500 ml)and the organic phase was washed Panso3and brine. The organic phase was dried and concentrated to obtain 4.9 g of the crude product. After adding petroleum ether to the solution of the crude product in ethanol and subsequent filtration of the obtained 2-(2-amino-thiazole-5-ylmethylene)-malonic acid diethyl ester (1.13 g, 18%).

b) 2-(2-Amino-thiazole-5-ylmethyl)-malonic acid diethyl ester

NaCNBH3(1.88 g, and 29.9 mmol) was added to a stirred solution of 2-(2-amino-thiazole-5-ylmethylene)-malonic acid diethyl ester (1.13 g; 4.2 mmol) in ethanol at 0°C. the pH of the solution counter is laravale adding to the solution a small amount of bromocresol green. Was added dropwise concentrated Hcl until then, until the solution became yellow. The ice bath was removed and the reaction mixture was stirred at room temperature for 5 hours. Was added water, and the product was extracted with CH2Cl2. The combined organic phase was dried and concentrated to obtain 2-(2-amino-thiazole-5-ylmethyl)-malonic acid diethyl ester (1 g, 87.8 per cent).

a) 2-(2-tert-Butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid diethyl ester

Vos2O (0.6 g, 27.5 mmol) was added to a solution of triethylamine (0.4 ml; to 30.1 mmol), 4-(dimethylamino)pyridine (0.34 g, 27.8 mmol) and 2-(2-amino-thiazole-5-ylmethyl)-malonic acid diethyl ester (0.75 g; 27.5 mmol) in CH2Cl2(5 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight. Additionally Vos2O (0.15 g; 0.7 mmol) was added at 0°and the reaction mixture was stirred at room temperature for 1 hour. Added CH2CL2and the organic phase was extracted with 0.3 M KHSO4and brine. The organic phase was dried and concentrated to obtain 0,78 g of the crude product. NMR showed that approximately 40% of 2-(2-amino-thiazole-5-ylmethyl)-malonic acid diethyl ester remained. Vos2Oh was added (0.6 g; 27.5 mmol) to a solution of the crude product (0,78 g), triethylamine (0.4 ml; 3.1 mmol), 4-(dimethylamino)pyridine (0.34 g; 27.8 mmol) in CH2Cl2(5 ml) at 0°C. the Reaction mixture was stirred at room temperature overnight and the procedure was repeated. The crude product (1 g) was purified flash chromatography (heptane/EtOAc; 1:1) and HPLC to obtain 2-(2-tert-butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid diethyl ester (184 mg, 17.9 per cent).

g) 2-(2-tert-Butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid monotropy ether

2-(2-tert-Butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid diethyl ester (158 mg; 0.43 mmol) was dissolved in ethanol (1 ml) and THF (0.5 ml), was added a solution of KOH (24 mg; 0.43 mmol) in ethanol (0,14 ml) at 0°C. the Reaction mixture was stirred at room temperature for 96 hours and then poured into ice water. The aqueous phase was extracted with diethyl ether, acidified to pH 3 by addition of 0.5 M HCl and was extracted with diethyl ether. The combined organic phase was dried and concentrated to obtain 2-(2-tert-butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid of monoethylene ester (97 mg, 66,4%).

d) 2-(2-tert-Butoxycarbonylamino-thiazole-5-ylmethyl)-acrylic acid ethyl ester

To a mixture of 2-(2-tert-butoxycarbonylamino-thiazole-5-ylmethyl)-malonic acid of monoethylene ester (94 mg; 0.27 mmol), 36% aqueous formaldehyde (36 μl; 1.2 mmol), CH2 CL2(0.2 ml) and water (0.2 ml) was added at 0°With diethylamine (30 μl; 0.40 mmol). The reaction mixture was stirred at room temperature overnight, poured into ice water and was extracted with CH2Cl2. The combined organic phases are washed with 5% NaHCO3, was dried and concentrated to obtain 2-(2-tert-butoxycarbonylamino-thiazole-5-ylmethyl)-acrylic acid ethyl ester (69 mg, 80.9 per cent).

e) 2-Acetylsalicylate-3-(2-tert-butoxycarbonylamino-thiazol-5-yl)-propionic acid ethyl ester

The triethylamine (32 μl, 0.23 mmol) was added to a solution of 2-(2-tert-butoxycarbonylamino-thiazole-5-ylmethyl)-acrylic acid ethyl ester (67 mg, 0.21 mmol) in teoksessa acid (0.4 ml) at 0°C. the Reaction mixture was stirred at room temperature for 48 hours and then poured into ice water. The aqueous layer was extracted with CH2Cl2. The combined organic phases were washed with saturated NaHCO3, was dried and concentrated to obtain 190 mg of the crude product. The crude product was purified flash chromatography (heptane/EtOAc, 1:0 68:32) to obtain 2-acetylsalicylate-3-(2-tert-butoxycarbonylamino-thiazol-5-yl)-propionic acid ethyl ester (43 mg, 51.6 per cent).

g) 3-(2-Amino-thiazol-5-yl)-2-mercaptomethyl-propionic acid

A solution of 2-acetylsalicylate-3-(2-tert-butoxycarbonylamino-thiazol-yl)-propionic acid ethyl ester (43 mg, 0.11 mmol) in concentrated HCl (1.5 ml) was boiled under reflux in an argon atmosphere for 1.5 hours. The reaction mixture was left to warm to room temperature and concentrated under reduced pressure to obtain 30 mg of the crude product. The crude product was purified preparative HPLC to obtain specified in the title compound (8 mg; 21%) in the form of cleaners containing hydrochloride salt.

1H NMR (500 MHz, D2O): δ 2.75-3.1 (m, 5H), 7.0 (br s, 1H).

MS (+) 219 (M+1).

Reduction

Ac = acetate

aq = water

AIBN = α,α’-azoisobutyronitrile

PG = benzyl

BU = butyl

Bz = benzoyl

DCC = dicyclohexylcarbodiimide

DIAD = diisopropylsalicylic

DIBAL = diisobutylaluminium

DIPEA = diisopropylethylamine

DMAP = N,N-dimethylaminopyridine

DME = 1,2-dimethoxyethane

DMF = dimethylformamide

DMSO = dimethyl sulfoxide

EDC = 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide

it = enantiomeric excess

Et = ethyl

EtOAC = ethyl acetate

EtOH = ethanol

h = hour

SPLA = acetic acid

HOBt= 1-hydroxybenzotriazole

HPLC = high performance liquid chromatography

KHMDS = bis(trimethylsilyl)amide and potassium

LDA = diisopropylamide lithium

MSRWA = 3-chloroperbenzoic acid

Me = methyl

Meon = methanol

min = minutes

RMV = 4-methoxybenzyl

Ph = phenyl

WG = cut

Rover (benzotriazol-1 yloxy)triprolidine hexaphosphate

Red-Al = bis-(2-methoxyethoxy)alumoweld sodium

TEA = triethylamine

TFA = triperoxonane acid

THF = tetrahydrofuran

Tos = toluene-4-sulfonyl

Example 35

In the analysis, described in “In vitro experiments for compounds according to the invention, which is described in the preceding examples, the following data were obtained on their ability to inhibit carboxypeptidase U:

pic50CPU at pH 7.4pIC50CPU at pH 7.4
Example (* = pH 8)Example (* = pH 8)
Example 15,5*Example 21/A5
Example 33,4*Example 21 B/W4
Example 43,3*Example 21/6,3
Example 55,5*Example 21/G4,5
Example 66,6Example 225
Example 76,2*Example 23a 4.9
Example 83,6*Example 245,8
Example 95,8Example 256
Example 10 2,7*Example 26of 5.4
Example 114,0*Example 27/A6,1
Example 123,9*Example Ba 4.9
Example 134,8*Example 27/6,1
Example 146Example 27/Gthe 5.7
Example 155,1Example 28the 5.7
Example 165,1Example 295
Example 175,9Example 305,2
Example 185,2Example 316,1
Example 20/A4Example 33the 5.7
Example 20 B/Wof 5.4Example 345,9
Example 20/6,4  
Example 20/G5  

As can be seen from the presented results, the compounds according to the invention are active inhibitors of carboxypeptidase u

Example 36

Compounds according to the invention can be represented, for example, in the form of the following drugs.

1. The solution for parenteral administration

Active connection (connected to the e of the invention) 5 g

Sodium chloride for injection 6 g

1 M sodium hydroxide to bring the pH up to pH 5-7

Water for injection to 1000 ml

The active compound and the sodium chloride is dissolved in water. the pH is brought to pH 5-7 by addition of an aqueous sodium hydroxide solution and then add water to the final volume. The resulting solution is sterilized by filtration and immediately poured into sterile ampoules of 10 ml, which is hermetically sealed.

2. Tablets for oral administration

Active connection (connection according to the invention) 150 g

The aluminosilicate of sodium, 20 g

Paraffin wax 120g

Microcrystalline cellulose 20 g

Hydroxypropylcellulose 5 g

Fumarate sodium 3 g

The active compound, sodium aluminosilicate, paraffin and microcrystalline cellulose are mixed and added an aqueous solution of hydroxypropylcellulose. The resulting mixture is dried and milled to obtain granules, which are then mixed with sodium fumarate. The mixture is pressed into tablets in a tablet machine to obtain 1000 tablets each containing 150 mg of active connections.

Compounds according to the invention can also be prepared in the form of a combined preparation combined, for example, with such an antiplatelet agent such as aspirin. An example of such a drug is the next drug.

3. Tablets is for oral administration (combined product)

Active connection (connection according to the invention) 100 g

Acetylsalicylic acid (aspirin) 100 g

Lactose 400 g

Polyvinylpyrrolidone 60 g

Microcrystalline cellulose (60 g

Magnesium stearate 12 g

Active connection, aspirin and lactose are mixed with an aqueous solution of polyvinylpyrrolidone. The mixture is dried and ground to form granules which are then mixed with microcrystalline cellulose and magnesium stearate. Then the obtained mixture is pressed into tablets in a tablet machine, receiving 1000 tablets each containing 100 mg of active compound and 100 mg of aspirin.

In addition, the preparations of the compounds according to the invention can be administered in conjunction with drugs antithrombotic agent with a different mechanism of action, for example in conjunction with the drug aspirin. In this case, these drugs can be represented as the following set of features:

4. The set of components

Carton containing blister packs of 10 tablets active compounds produced in accordance with the note (2), and blister packs of 10 tablets of aspirin 100 mg, manufactured in the traditional way.

1. The compound of General formula I

or its pharmaceutically acceptable salt or MES or MES such salts, where

R1represents a

With1-C6alkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

aliphatic heterocyclyl containing at least one nitrogen atom, possibly substituted by one or more than one basic group such as amino, amidino and/or guanidino;

aromatic heterocyclyl containing at least one nitrogen atom, substituted by one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

or aryl substituted by one or more than one basic group such as amino, amidino and/or guanidino;

R2represents H, alkyl, alkylthio, alkoxy or cycloalkyl;

R3represents COOR5, SO(OR5), SO3R5P=O(OR5)2B(OR5)2P=OR5(OR5or tetrazol, or ISOStAR carboxylic acid, which represents an acid group having a pKa from about -5 to about 25;

R4represents SH or S-WITH-C1-C6alkyl;

R5represents N or C1-C6alkyl;

R6represents N;

X represents C(Z)2or NR6CO;

Y represents C(Z)2;

Z independently represents H, C1-C6alkyl, aryl or cycloalkyl.

2. The compound according to claim 1 or its pharmaceutically acceptable salt or MES or MES such salts, where

R1represents a

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

aliphatic heterocyclyl containing at least one nitrogen atom, possibly substituted by one or more than one basic group such as amino, amidino and/or guanidino;

aromatic heterocyclyl containing at least one nitrogen atom, substituted by one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

or aryl substituted by one or more than one basic group such as amino, amidino and/or guanidino;

R2depict is to place a N, alkyl, alkylthio, alkoxy or cycloalkyl;

R3represents COOR5;

R4represents SH or S-CO-C1-C6alkyl;

R5represents N or C1-C6alkyl;

R6represents N;

X represents C(Z)2;

Y represents C(Z)2;

Z independently represents H, C1-C6alkyl, aryl or cycloalkyl.

3. The compound according to claim 1 or 2, or its pharmaceutically acceptable salt or MES or MES such salts, where

R1represents a

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

aliphatic heterocyclyl containing at least one nitrogen atom, possibly substituted by one or more than one basic group such as amino, amidino and/or guanidino;

aromatic heterocyclyl containing at least one nitrogen atom, substituted by one or more than one basic group such as amino, amidino and/or guanidino;

heterocyclyl containing at least one heteroatom selected from S or O, and substituted one or more than one basic group such as amino, amidino and/or guanidino;

R2represents N or C1 -C3alkyl;

R3represents COOR5;

R4represents SH or S-CO-C1-C6alkyl;

R5represents N or C1-C6alkyl;

X represents C(Z)2;

Y represents C(Z)2;

Z independently represents N or C1-C6alkyl.

4. The compound according to any one of claims 1 to 3, or its pharmaceutically acceptable salt or MES or MES such salts, where

R1represents a

cycloalkyl, substituted one or more than one basic group such as amino, amidino and/or guanidino;

aliphatic heterocyclyl containing at least one nitrogen atom, possibly substituted by one or more than one basic group such as amino, amidino and/or guanidino;

aromatic heterocyclyl containing at least one nitrogen atom, substituted by one or more than one basic group such as amino, amidino and/or guanidino;

R2represents N or C1alkyl;

R3represents COOR5;

R4represents SH or S-CO-C1-C6alkyl;

R5represents N or C1-C6alkyl;

X represents C(Z)2;

Y submitted is a C(Z) 2;

Z independently represents N or C1-C6alkyl.

5. The compound according to any one of claims 1 to 4, or its pharmaceutically acceptable salt or MES or MES such salts, where

R1represents a

cyclopentyl, pyridyl, pyrimidinyl, piperidinyl or thiazolyl;

R2represents N or C1alkyl;

R3represents COOR5;

R4is a SH;

R5represents N;

X represents CHZ;

Y represents CHZ;

Z independently represents N or C1-C6alkyl.

6. A method of obtaining a compound according to any one of claims 1 to 5, where R1, R3, R4and Y are such as defined in claim 1, X represents C(Z)2and R2represents H, at which

the compound of formula VI

where R1, R3and Y are such as defined in claim 1 and X represents a C(Z)2, is subjected to the interaction with the compound of the formula IX

where R5is a suitable protecting group such as acetyl (AC), benzoyl (Bz), 4-methoxybenzyl (RMV) or benzyl (EAP), one or in the presence of a suitable base, such as NaOMe, NaH or triethylamine, or, alternatively, prisutstvie free-radical initiator, such as α,α’-azoisobutyronitrile (AIBN), under standard conditions.

7. A method of obtaining a compound according to any one of claims 1 to 5, where R1, R2, R3and R4such as defined in claim 1, Y is CH2and X represents a C(Z)2in which

the compound of formula XIV

where R1, R2and R3such as defined in claim 1 and X represents a C(Z)2, is subjected to the interaction with the compound of General formula IX

where R5is a suitable protecting group, such as AC or Bz, in the presence of a suitable reagent such as triphenylphosphine/diisopropylsalicylic (PPh3/DIAD), under standard conditions.

8. A method of obtaining a compound according to any one of claims 1 to 5, where R1, R2, R3, R4and Y are such as defined in claim 1, and X represents NR6CO., in which the compound of General formula XV

where R2, R3, R6and Y are such as defined in claim 1, a R5is a suitable protecting group, such as Ac, Bz, PMB, or EAP, is subjected to the interaction with the compound of General formula XVI

where R1such as defined in claim 1 and X represents COOH, prisutstvie suitable binding reagents, such as (benzotriazol-1 yloxy)triprolidine hexaflurophosphate/diisopropylethylamine (PyBOP/DIPEA), dicyclohexylcarbodiimide/1-hydroxybenzotriazole (DCC/HOBt), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide/triethylamine/N,N-dimethylaminopyridine (EDC/TEA/DMAP) or pyridine, in standard conditions.

9. Pharmaceutical drug that has inhibitory activity against carboxypeptidase U and containing a compound according to any one of claims 1 to 5 as an active ingredient in combination with pharmaceutically acceptable adjuvant, diluent or carrier.

10. The compound according to any one of claims 1 to 5 for the manufacture of drugs for treatment or prevention of conditions associated with inhibition of carboxypeptidase U.

11. The compound according to any one of claims 1 to 5 for the manufacture of drugs for the inhibition of carboxypeptidase U.

12. Pharmaceutical formulation for use in the treatment or prophylaxis of conditions associated with inhibition of carboxypeptidase U, containing the compound according to any one of claims 1 to 5 in combination with a pharmaceutically acceptable adjuvant, diluent or carrier.

13. Pharmaceutical drug that has inhibitory activity against carboxypeptidase U and containing

1) the compounds of formula I or its pharmaceutically acceptable salt or MES or MES of such salt, and

2) one or more is it than one antithrombotic agent with a different mechanism of action, such as antiplatelet agent, an inhibitor of thromboxane receptor, the synthetase inhibitor, a receptor antagonist of fibrinogen, a prostacyclin mimetic, a phosphodiesterase inhibitor or receptor antagonist (P2T) adenosine-5’-diphosphate (ADP-receptor (R2T)

in a mixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

14. The set of components having inhibitory activity against carboxypeptidase U, including

1) the pharmaceutical preparation containing the compound of formula I or its pharmaceutically acceptable salt or MES or MES such a salt, in a mixture with a pharmaceutically acceptable adjuvant, diluent or carrier; and

2) pharmaceutical preparation containing one or more than one antithrombotic agent with a different mechanism of action, such as antiplatelet agent, an inhibitor of thromboxane receptor, the synthetase inhibitor, a receptor antagonist of fibrinogen, a prostacyclin mimetic, a phosphodiesterase inhibitor or antagonist of ADP-receptor (R2T)

in a mixture with a pharmaceutically acceptable adjuvant, diluent or carrier;

moreover, the compound (1) and agent (2) is presented in forms that are suitable for administration in conjunction with each other.



 

Same patents:

The invention relates to derived-alanine General formula I:

in which Q1or Q3independently from each other represent CH or N, they cannot mean N; R1represents H, A, Ar or Hal, R2denotes H or A,

each of R4and R5independently denotes H, a or Hal, R6represents H or a, a denotes alkyl with 1-6 carbon atoms, Ar is an unsubstituted aryl, Hal denotes F, Cl, Br or I, n is 2, 3, 4, 5 or 6, m is 1, 2, 3 or 4, as well as their physiologically acceptable salt and solvate; two methods for their preparation and pharmaceutical product having the properties of integrins inhibitor

The invention relates to new derivatives of 2-(O-[pyrimidine-4-yl]metalinox)phenylacetic acid, their salts and N-oxides of the General formula I, means for combating harmful fungi and pests, the method of production thereof and method of combating harmful fungi and pests with the use of compounds of General formula I

The invention relates to new derivatives of guanidine and their pharmaceutically acceptable salts, used as medicines

The invention relates to applicable in medicine new derived aminotriazole or its hydrate and its pharmaceutically acceptable salts

The invention relates to the class of pyrimidine compounds, which are suitable for the treatment of diseases and disorders of the Central nervous system (CNS), for example to prevent cerebral ischemic lesions, containing their pharmaceutical compositions and to methods for their preparation

The invention relates to new derivatives of acrylic acid, exhibiting fungicidal activity mainly in relation to fungal infections of plants, processes for their preparation, fungicidal compositions and method of combating fungi

The invention relates to heterocyclic compounds having angiotensin II antagonistic activity

The invention relates to new derivatives of benzene or pyridine of the formula (I)

where R denotes H, C1-C7alkyl and halogen; R1denotes H or halogen, provided that in the 4th position R1not denotes bromine or iodine; R2denotes H or CF3; R3denotes N or C1-C7alkyl; R4denotes H, halogen, C1-C7alkyl and others; R5denotes N or C1-C7alkyl; X represents-C(O)N(R5)-, -N(R5)-C(O)- or-C(O)O-; Y represents -(CH2)n-, -O-, -S-, -SO2-, -C(O)- or N(R5’)-; R5’means (ness.)alkyl; Z represents =N-, -CH= or-C(C1)=; n denotes a number from 0 to 4; and their pharmaceutically acceptable salts

The invention relates to new derivatives of 3-phenylpyridine formula (I)

in which R denotes hydrogen, C1-C7alkyl, C1-C7alkoxy, halogen or CF3, R1means H or halogen, or R and R1may together form-CH=CH-CH=CH-; R2means H, halogen, CF3, R3means H or C1-C7alkyl, R4means H or piperazine-1-yl; R5means H or C1-C7alkyl, X is-C(O)N(R5)-, -(CH2)mO-, -(CH2)mN(R5)-, -N(R5)C(O)- or N(R5)(CH2)m-; n is an integer from 0 to 4, m is 1 or 2, and their pharmaceutically acceptable acid additive salts

The invention relates to new salts of pyridinium General formula (I) or their pharmaceutically acceptable salts, where R1is-R4- R5or-N(R7)N(R7R9, R4choose from the group of-N(R7R6O-, N(R7R6N(R7), -OR6O-,

-OR SIG6N(R7)-, where R6- alkyl, R5choose from the group of alkyl, aryl, including heteroaryl, -COR7, -SO2R7and-COR10where R7Is H, alkyl or aryl, including heteroaryl, R2Is F, Cl, Br, J, alkyl, aryl, including heteroaryl, formyl, acyl, C(O)NR7R10or C(O)or SIG7, m = 0, 1, or 2, R3selected from the group comprising R7OR7N(R7)(R10) and CH(R7)C(O)R8, R8is R7OR7and NR7R10, R9is hydrogen, alkyl, aryl, including heteroaryl, -C(O)R10, -SO2R10, -C(S)OTHER10, -C(NH)NH(R10), -C(O)OTHER10, R10- H, alkyl, or aryl, including heteroaryl, and in each case, it is not necessarily different from R7X represents an ion halogen provided that 1) when two alkyl groups are the same carbon or nitrogen, they are not necessarily linked together with the formation of a cyclic structure, and (2) nitrogen heteroaryl ring R1

The invention relates to new Amida acids of the formula I

< / BR>
where R1- C1-C6alkanoyl,1-C6alkoxycarbonyl, benzoyl, benzoyl substituted halogen (C1-C6)-alkoxy, C1-C6alkylsulfonyl, phenylsulfonyl, phenylsulfonyl, substituted with halogen, or cyclo (C3-C6) alkylsulphonyl, R2- phenyl, phenyloxy or phenylamino, where each phenyl may be substituted with halogen; pyridyl or pyridylamino, a represents a single bond, E is ethylene, X represents CH, Y is-NR5where R5is hydrogen, Q is-C(O)- or-SO2-, R3and R4together form ethylene, or their pharmaceutically acceptable salts

The invention relates to new pyridinecarboxamide formula (I)

< / BR>
or their tautomeric forms, where the join pyridine in position 2, 3 or 4; R1denotes hydrogen; Q represents C1-C9is a divalent saturated hydrocarbon radical; X denotes methylene or oxygen; R represents hydrogen, halogen, nitro, methylenedioxy, alkyl or alkoxy containing from 1 to 4 carbon atoms, or their pharmaceutically acceptable non-toxic smolem and N-oxides

The invention relates to an inhibitor of the activity of esterified cholesterol transport protein (HETB), comprising as active ingredient a compound represented by the formula (I), where R is a straight or branched alkyl group; a lower halogenating group; substituted or unsubstituted cycloalkyl group; substituted or unsubstituted cycloalkylcarbonyl group; substituted or unsubstituted aryl group, or substituted or unsubstituted heterocyclic group, X1X2X3and X4may be the same or different and each represents a hydrogen atom, halogen atom, lower alkyl group, lower halogenating group; a lower alkoxygroup; a cyano; a nitro-group; Y represents-CO -, and Z represents a hydrogen atom or mercaptohexanol group, or its pharmaceutically acceptable salt, or hydrate, or MES

The invention relates to a new process for the preparation of 2,3-pyridinecarboxamide formula (I), where R is hydrogen, C1-C6alkyl or C1-C6alkoxymethyl, R1means hydrogen, C1-C6alkyl, C(O)R2, phenyl, benzyl, R2means C1-C6alkyl, benzyl or phenyl, which is that the compound of formula (II), where R has the above meaning, R6means C1-C6alkyl, R7means OR8or NR9R10, R8means hydrogen, C1-C6alkyl, C(O)R11, phenyl, benzyl, R11means C1-C6alkyl, OR12, NR12, R13, benzyl or phenyl, R12and R13denote hydrogen, C1-C6alkyl, benzyl or phenyl, R9and R10denote hydrogen, C1-C6alkyl, benzyl or phenyl, is subjected to the interaction with the imide of maleic acid of the formula (III), where R1have the above values

The invention relates to the class of hydrazides of dicarboxylic acids, namely to a new biologically active isonicotinohydrazide tarakanovas acid formula

< / BR>
The compound obtained by the interaction of the hydrazide of isonicotinic acid anhydride citramonum

The invention relates to new derivatives of 2-aminopyridine of General formula (I)

where And denotes the radical

in which R1, R2and R3mean hydrogen, halogen, HE, alkyl or alkoxy and the other,

or radical

in which R8means hydrogen, x is the radical -(CH2)m-Q, m is an integer from 0 to 6, Y represents alkyl, alkenylphenol or alkenylphenol chain or other, R10is hydrogen or alkyl, or their salts
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