Derivatives nitroalkanes acids or their pharmaceutically acceptable salts

 

(57) Abstract:

Usage: in medicine, in the treatment of heart disease. The essence of the invention: derivatives nitroalkanes acids of the formula where R1- lower AlKO attached peptide bond Pentapeptide: Tyr-Pro-Phe-Pvo-GLy(OMe); R1= H, Alk(C1-C6) , acylamino; R2= H, lower Alk; R3= H; R4= H, lower Alk; R5=(CH2)kSX , where X = H, lower Alk or X = COR , where R = H, lower Alk or N - acylaminoalkyl; m = 0.1 to 10; n and O = 0 to 2, or their pharmaceutically acceptable salts. Reagent I : where R6- alkylthiol. Reagent II : amino acid and/or N - acylaminoacyl, peptide or N - elliptic with 2 - 5 attached peptide bond amino acid residue with the formation of tiefer. Compounds have low toxicity. table 2.

The invention relates to a derivative nitroalkanes acids or their pharmaceutically acceptable salts.

Organic nitrates (esters of nitric acid) has been proven in the treatment of heart disease.

Their effect is manifested not only in the unloading of the heart by lowering pre-and post loads, but also in the increased supply of oxygen to the heart due to expansion kanceska nitrates, such as glyceryl trinitrate (TNG), isosorbide-5-Mononitrate or isosorbide-dinitrate treatment, being continuously introduced into the body in large quantities within a short period of time lose a significant portion of its actions, i.e., there is the addiction to these substances. The results of numerous experiments indicate that the presence of sulfhydryl groups may hinder the development process of accustoming to nitrates and slow down already beginning the process of habituation.

Currently under development mechanism of habituation refers to the following.

According to the modern level of knowledge of the pharmacological action of organic nitro compounds depends on the presence of cysteine. With him organic nitrate forms an intermediate product from which due to decay along with other products are released NO radicals, which in turn activate the target enzyme is soluble guanilatziklazu smooth muscle cells. Finally, the subsequent reaction caused by the formation of GMP, lead to a relaxation of tension and vasodilation.

Assume that the above-mentioned short-lived reactive intermediate product prerogrative and subsequent, not yet clarified, reactions formed nitrosothiol, which released nitric oxide or nitrite ions. On the other hand, suggest that the enzymatic decomposition using GSH-reductase, leading exclusively to the formation of nitrite ions, plays no role in the manifestation of pharmacological action. So, as above described, for non-enzymatic decomposition of the intermediate product is essential cysteine, and this decay depending on the dose, subject to depletion (depletion of sulfhydryl groups), so that over time decreases the amount of the released radicals NO, activates guanilatziklazu that clinically is manifested in the decrease of pharmacological action.

In the European patent N 0362575 described in connection with specific structure composed of nitration acids (nitroalkanes acids), serosoderjaschei amino acids or peptides.

The presence of sulfhydryl groups is intended to prevent or delay the development process of accustoming to nitrates.

In the cited patent is described, for example, compounds containing such sulfur-containing amino acids like cysteine or methionine, in the form of their mitrovska alkenylboronic acid number of atoms With 2-8.

Although these compounds have valuable pharmacological properties, in the sense that they hinder the development process of accustoming to nitrates and slow already beginning the process of addiction, they are characterized by certain disadvantages. They have a low melting point, do not dissolve in water and difficult to obtain in pure form.

Thus, the present invention consists in obtaining and providing specialist new organic compounds that do not have the aforementioned disadvantages.

The objective of the invention is solved by the fact that the proposed compounds are derived nitroalkanes acids of General formula

O2NOCH(CH2)(CH2)(CH2R

(I) where R is lower alkoxy or attached peptide bond Pentapeptide Tyr-Pro-Phe-Pro-Gly (OMe);

R1is hydrogen, alkyl (C1-C6), acylamino;

R2is hydrogen, lower alkyl;

R3is hydrogen;

R4is hydrogen, lower alkyl;

R5- (CH2)kSX, where X is hydrogen, lower alkyl or X-CORIIin which RIIis hydrogen, lower alkyl or a residue of N-acylaminoalkyl;

m is 0-10, n and o is 0-2, or their pharmaceutically acceptable salts.

Offer protirelin such amino acids, as cysteine, methionine or homocysteine.

According to another variant implementation of the invention, the amino acids are stereochemically L-form.

Sulfur-containing amino acids on the C-end can be tarifitsirovana.

According to a preferred variant implementation of the invention, amino acids - cysteine and/or methionine, can be in the form of methyl, ethyl or propyl esters.

Particularly preferred

ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine,

ethyl ester of N-NITROPHENOL-S-(N-acetylethyl)-L-cysteine,

ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine.

The proposed compounds of formula I get famous due to the fact that the compound of General formula O2NOCH(CH2)(CH2)(CH2R ,

(II) where R1-R4, m, n, o have the above meanings;

R6- lower alkylthio are known for the formation of tiefer, i.e. reactions with amino acids, N-acylaminoalkyl, peptides or N-ellipticity with 2-5 attached peptide bond amino acid residues. The reaction for obtaining compounds of General formula II is carried out, for example, as described in European patent N the above values can be used in the form of their free acids, reactive halides, azides, esters and anhydrides, and subjected to reaction with a compound of General formula

H(CH2)(CH)R , where R, R3, R4, R6, n and o have the above meanings and contain amino acids and/or peptides, resulting in the formation of compounds of General formula II.

For translation of the compounds of General formula I, their pharmacologically compatible salts they are subjected to the reaction, preferably in an organic or aqueous organic solvent with an equivalent amount of inorganic or organic acids, for example hydrochloric, Hydrobromic, nitric, phosphoric, sulfuric, formic, acetic, propionic, oxalic, fumaric, maleic, succinic, adipic, benzoic, salicylic, o-acetoxybenzoic, cinnamon, naphthoic, almond, lemon, malic, tartaric, aspartic, glutamic, methanesulfonic or n-toluensulfonate.

We offer new compounds of General formula I and their salts can be entered enterally or parenterally in liquid or solid form.

As the environment for the introduction of drugs preferably use water containing commonly used in injectable solutions of the additive, the feats of buffers, ethanol, complexing agents (such as ethylenediaminetetraacetic acid and its non-toxic salt), high-molecular polymers (such as liquid polyethylene oxide), used to control viscosity. Solid carriers include starch, lactose, mannitol, methylcellulose, talc, highly dispersed silica, high molecular weight fatty acids (such as stearic), gelatin, agar-agar, calcium phosphate, magnesium stearate, animal and vegetable fats and solid high molecular weight polymers (such as polyethylene glycols). Suitable for oral administration of drugs may contain flavoring and sweetening agents.

According to another variant implementation of the invention, medicines contain a certain amount of one of the proposed compounds and/or their mixture.

Such drugs can be used for the treatment of cardiovascular diseases, such as koronrodilatirtee funds, funds for the treatment of hypertension, heart failure and expansion of peripheral blood vessels, including vessels of the brain and kidneys.

Pharmaceutical preparations containing a predetermined amount of one or more of the proposed connection, you can itromid time. Daily doses beginning of the current amount of 20-300 mg, assuming a body weight of 75 kg Proposed connection you can also enter 1-8 times per day in the form of injections or by continuous intravenous infusion, and is usually sufficient amount of 5-200 mg/day.

A typical tablet may have the following composition mg:

1) Ethyl ester of N-Nitra-

topiwale-S-(N-acetyl - glycyl)-L-cysteine 25 2) Starch, USP* 57 3) Lactose, USP* 73 4) Talc, USP* 9 5) Stearic acid 6

*USP - pharmaceutical purity according to the USP monograph

Substances 1, 2 and 3 sift through a sieve, granularit and then mixed with substances 4 and 5 to achieve a homogeneous mixture. The mixture is then pressed into tablets.

The examples illustrate the invention without limiting it.

P R I m e R 1. Obtaining the ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine.

48 g (0.41 mol) of N-acetylglycine with stirring at room temperature weigh in 300 ml of methylene chloride (CH2Cl2), and then cooled to 10aboutC. With stirring, a solution of 109,8 g (0,373 mol) of ethyl ester of N-NITROPHENOL-L-cysteine in 300 ml of CH2Cl2moreover , the reaction proceeds poorly exothermically. Reacts add to 84.6 g (0.41 mol) of dicyclohexylcarbodiimide (DCC) in 200 ml of CH2Cl2so that the temperature was within 5-10aboutC. After heating the mass to room temperature it is stirred for 4 days at room temperature. Suck urea - DCK and washed with CH2Cl22 times in 100 ml.

Collected methylenchloride phase is successively washed with 200 ml of a 9% aqueous sodium bicarbonate solution, 300 ml of 1N. solution of hydrogen chloride and 300 ml of dist. water. Finally methylenchloride phase is dried with anhydrous sodium sulfate and concentrated on a rotary evaporator type of the rotavapor Buchi company until constancy of weight.

The result 162,9 g (calculated output - 146,7 g) of the target product as a pale yellow oil.

162,9 g of the target product at room temperature is dissolved in 470 ml of ethyl acetate. After stirring for 15 min at room temperature, filtered formed insoluble white precipitate. Then to the clear light yellow filtrate at room temperature with stirring, slowly add 390 ml of n-hexane.

To the resulting solution was added seed crystals and stirred over night at room temperature. The precipitated crystals are sucked off and at room temperature for 2 times about chumney dryer at room temperature and a vacuum of 2 Torr.

The yield of the target product is of 85.4 g (calculated output 146,74 g).

Sopl.71,8aboutC.

P R I m m e R 2. Obtaining the ethyl ester of N-NITROPHENOL-S-(N-acetylethyl)-L-cysteine.

53,8 g (0.41 mol) of N-acetylamino with stirring at room temperature weigh in 300 ml of methylene chloride (CH2Cl2), and then cooled to 10aboutC. With stirring, a solution of 109,8 g (0,373 mol) of ethyl ester of N-NITROPHENOL-L-cysteine in 300 ml of CH2Cl2moreover , the reaction proceeds poorly exothermically. The reaction mass with stirring, cooled to 5aboutAnd then, continuing to mix, slowly added dropwise to 84.6 g (0.41 mol) of dicyclohexylcarbodiimide (DCC) in 200 ml of CH2Cl2so that the temperature was within 5-10aboutC. After heating the mass to room temperature it is stirred for 4 days at room temperature. Suck urea - DCK and washed with methylene chloride 2 times in 100 ml.

Collected methylenchloride phase is successively washed with 200 ml of 9% solution NaHC3, 300 ml of 1N. HCl solution and 300 ml of dist. water. Finally methylenchloride phase is dried with anhydrous sodium sulfate, and concentrate on rotationand 151,84 g) of the target product as a pale yellow oil.

160,5 g of the target product at room temperature is dissolved in 345 ml of ethyl acetate. After stirring for 15 min at room temperature, filtered formed insoluble white precipitate. Then to the clear light yellow filtrate at room temperature with stirring, slowly add 345 ml n-hexane.

To the resulting solution was added seed crystals and stirred over night at room temperature. The precipitated crystals are sucked off and at room temperature twice washed with 100 ml of a mixture of 20 ml of ethyl acetate and 80 ml of n-hexane.

The crystals are dried to constant weight in a vacuum dryer at room temperature and a vacuum of 2 Torr.

The yield of the target product is of 78.2 g (calculated output 151,84 g).

Sopl.76,6aboutC.

P R I m e R 3. Obtaining the ethyl ester of N-NITROPHENOL-S-(N-acetyl-leucyl)-L-cysteine.

0.02 mol (6 g) containerbase ether NITROPHENOL acid dissolved in 100 ml of dichloromethane. At 10aboutWith the supply of nitrogen is added slowly to 0.03 mol (5,19 g) N-acetyl-leucine and 0.1 g of dimethylaminopyridine (DMAP). Then added dropwise 0.03 mol (6,15 g) dicyclohexylcarbodiimide (DCC), ASS="ptx2">

Processing.

The above mixture is sucked off. The solution is successively extracted with equivalent quantities of 0.1 n solution of hydrogen chloride, saturated solution of sodium bicarbonate and SW. water. Then on a rotary evaporator type of the rotavapor Buchi company remove the solvent. Remains 10 g of oily residue.

Recrystallization.

10 g of the above oily substance, slightly warming, dissolved in 45 ml of ethanol and 40 ml of water. Then the resulting substance during the night vykristallizovyvalas in the refrigerator. The obtained crystals are sucked off and dried in a vacuum dryer.

The mass spectrum of the substance confirms its structure. Melting point: 91,4aboutC. Analysis BIH: 98,7%. Yield 5 g = 0.012 mol =

= 57,4% of theoretical-

ical.

P R I m e R 4. Obtaining L-methioninamide ether 2-nitroxyethyl acid.

10.6 g of the Foundation of the ethyl ester of L-methionine by 60.0 mmol

8,9 g 2-nitrosoanabasine acid 60,0 mmol

13,0 g 98% (DCC) (dicyclohexylcarbodiimide) is 61.5 mmol

20 mg DMA (dimethylaminopyridine)

185 ml of CH2Cl2< / BR>
Testing and processing carried out as in ethyl ether N-(2-nitrosobutane acid as a pale yellow oil = 111,0% of theory.

From the crude product L-methioninamide ether 2-nitroxyethyl acid get solution of 50 mg/ml methanol for analysis thin-layer chromatography. The applied volume of 1 ál = 50 micrograms of the substance. Conducting thin-layer chromatography and conditions are consistent with those used for the ethyl ester of N-(2-nitrosobutane)-L-cysteine in an atmosphere of nitrogen.

The result of thin-layer chromatography.

The crude product contains about 10% of impurities containing-O-NO2. Purification of the crude product is carried out by chromatography on a column according to the MPLC system (SIH) company Buechi: 20,5 g 2-methioninamide ether 2-nitroxyethyl acid dissolved in 51,5 ml of methanol and 20 ml of distilled water. The filtered solution is then applied to the column.

Concentrated 3. Faction:

13.8 g of ethyl ester of 2-methionine 2-nitroxyethyl acid (in the form of a colourless oil) = 74.6% of theory.

Concentrated 4. Faction:

2,95 g L-methioninamide ether 2-nitroxyethyl acid (in the form of a colourless oil) = 15.9% from Teoret.

P R I m e R 5. Getting thiopropionate ethyl ester nitrosophenol acid L-cysteine.

10.3 g of ethyl ester

L-cvetelina D = =0,73 (6,80 ml) is 49.0 mmol

60 ml of CH2Cl2< / BR>
Testing and processing carried out as in S-carbonate ethyl ester of N-(3-NITROPHENOL)-cysteine (always in an atmosphere of N2).

Output 12,46 g N-Piv-Cy-Et-S-Prop in the form of a light brown oil = 101,6% of theory.

The result of mass spectrometry: confirmation of identity.

P R I m e R 6. Getting diisobutyrate ethyl ester of L-cysteine nitrosophenol acid.

10.3 g of ethyl ester

L-cysteine nitroxide - valinovoi acid 35,0 mmol

6.7 g of the anhydride of somas - lanai acid 42.0 mmol

5.0 g of triethylamine D = =0,73 (6,80 ml) is 49.0 mmol

60 ml of CH2Cl2< / BR>
Testing and processing carried out as in ethyl ether N-(2-nitrosobutane)-L-cysteine (always in an atmosphere of N2).

Output 13,39 g N-Piv-Cy-Et-S-i-But in the form of a light brown oil = 105,0% of theory.

The resulting mass spectrum: confirmation of identity.

P R I m e R 7. Obtaining the ethyl ester of cysteine 2-nitroglycerol acid.

Original mix:

0,015 mol = 2,74 g 2-nitroglycerol acid

0,015 mol = 2,78 g of ethyl ester of cysteine x HCl

1 volume spatula DMA dissolved in 50 ml of dioxane, PI in 20 ml of dioxane. The supply of N2.

Get a clear solution, from which, after cooling, precipitation.

Processing.

The solution is sucked off and is subjected to rotation processing, it is mixed with 100 ml of water and extracted with 100 ml of ethyl acetate with the supply of N2, the solution is subjected to rotation processing.

Output to 5.58 g (Calculated output 4,63 g).

The recrystallization perform 50 ml ETON

The output is about 60 mg.

P R I m e R 8. Obtaining the ethyl ester of cysteine 2-nitroxylenes acid.

Original mix:

0,033 mol = 5 g nitroxylenes acid

0.046 mol = 8,59 ethyl ester of cysteine x HCl

1 volume spatula DMA (dimethylaminopyridine) when heated, stirred in 100 ml of dioxane and dissolved by filing N2.

Dissolve 0.046 mol = 9,49 g DCC (dicyclohexylcarbodiimide) in 80 ml of dioxane and dropwise added slowly. Stirred over night.

The solution is sucked off and is subjected to rotation processing, to the residue water is added and when the salting out is extracted with ethyl acetate. The solution azeotropically is subjected to rotation processing.

Output 8,03 g (Calculated output 9,24 g).

P R I m e R 25 mol = 7 g of 3-N-But-Cy-Et slowly dissolved in 150 ml of CH2Cl2add 0.3 mol = 27.9 ml AU2Acting Upon the filing of N2and at a temperature of 5-10aboutWith dropwise added slowly to 0.3 mol = 30,4 g of triethylamine dissolved in 100 ml of CH2Cl2. Stirred over night.

Washed with a solution of an equivalent amount NaHC3and then water. The solution is carefully subjected to the rotation processing.

The output of 9.8 g

Will recrystallized from 50 ml of EtOH.

Sucked off the crystals.

Output in the dry state of 3.13 g = 38,84% of theory.

P R I m e R 10. Getting nitrato-Piv-CyEt-S-benzoate.

Receive:

Dissolve 0,034 mol = 10 g N-Piv-CyEt in 100 ml of CH2Cl2at a temperature of 5-10aboutAnd when applying N2then slowly add 0,0408 mol = 4.8 ml of benzoyl chloride, and then dropwise added slowly 0,034 mol = 4,7 ml (Et)3N, dissolved in 50 ml of CH2Cl2. Stirred for 3 days.

Processing:

The original mixture is poured on ice water, extracted it and put its rotary processing. Carry out purification by chromatography on a column.

The solvent 75 Meon, 25 N2O.

Fraction is subjected to rotation processing.

Thin-layer items the ATA 3-N-Piv-CyEt.

Dissolve is 0.023 mol 7 g N-Piv-CyEt in 100 ml of CH2Cl2at a temperature of 5-10aboutAnd when applying N2then dropwise added slowly 0,027 mol = 2,63 ml of ethyl ether of Harborview acid, and then added dropwise 0,023 mol = 3.2 ml (ET)3N. Stirred over night.

Processing: the solution serves to water and mix thoroughly, it is extracted and subjected to its rotary processing. Put in the fridge.

Output 9.3 g; Tpl.36,2aboutC.

P R I m e R 12. Obtaining N-(3-NITROPHENOL)-homocysteinaemia.

O-O-< / BR>
The number of initial mixture of 5.0 g of 0.03 mol Nitrative-

Lanovoy acid, 4.7 g of 0.03 mol of Homocysteine-

Alecto x HCl 2.4 g of 0.03 mol of Pyridine 6.2 g of 0.03 mol DCK 70 ml THF

5 g NITROPHENOL acid and 4.7 g of homocysteinaemia x HCl suspended in 50 ml of THF. After adding 2.4 g of pyridine at a temperature of 5-10aboutWith added dropwise a solution consisting of 6.2 g DCC and 20 ml THF. The initial mixture is additionally mixed at room temperature overnight and then filtered. The filtrate is subjected to rotation processing, and the residue is stirred with 150 ml of 1N. NaOH and extracted twice his 100 ml of CH2Cl2 is A.

The crude product is recrystallized from 20 ml of n-hexane and 10 ml EtOH. Yield 510 mg (approximately 6.5% of theory.).

P R I m e p 13. Obtaining the ethyl ester of N-(2-nitropropionic)-L-cysteine. < / BR>
The number of initial mixture:

26,9 g 0.20 mol nitromalonic acid

28.6 g 0,19 mol ethyl ester of cysteine

41,3 g 0.20 mol DZK

375 ml of CH2Cl2< / BR>
26,9 g nitromalonic acid and 28.6 g of ethyl ester of cysteine dissolved in 300 ml of CH2Cl2. At a temperature of 15-25aboutWith added dropwise a solution consisting of 41,3 g DCC and 75 ml of CH2Cl2. The initial mixture is additionally stirred overnight and then filtered. The filtrate is completely subjected to the rotation processing. The remainder of 50.5 g Of 100 ml EtOH will recrystallized to 18.5 g of crude product. Output 9.2 grams (approximately 49,64% of theory.).

P R I m e R 14. Obtaining the ethyl ester of N-(3-nitrohexane)-L-cysteine. H3COH< / BR>
Original mix: 5,1 g 0,029 mol 3-nitrates-

canovai acid 5,94 g to 0.032 mol hydrochloride

ethyl EPE-

RA cysteine to 6.58 g to 0.032 mol DCC Cat. DMA 70 ml of dioxane

to 5.1 g of nitrohexane acid, 5,94 g of the hydrochloride of the ethyl ester of cysteine and catalytic amounts DMA suspended in 50 ml DIAC is, return it within two days was stirred at room temperature and then filtered. Remove dioxane, add the remainder to 100 ml of AcOEt and extracted twice his 100 ml 1N. HCl. The organic phase is subjected to rotation processing. The remainder of 4.6 g (51,44% of theory.).

P R I m e R 15. Obtaining the ethyl ester of L-cysteine N-(2-nitrotoluene acid). O

Original mix: 11,35 g 0,094 mol 2-nitrococcus-

Noah acid

(dissolved

in 200 ml of AcOEt) 12,60 g of 0.085 mol of ethyl EPE-

RA cysteine 19,40 g 0,094 mol DCC 200 ml of AcOEt 20 ml of CH2Cl2< / BR>
Dissolve of 12.6 g of ethyl ester of cysteine in solution, consisting of 11,35 g 2-nitrotoluene acid and 200 ml of AcOEt. At a temperature of 20-25aboutWith added dropwise a solution consisting of 19,4 g DCC in 20 ml of CH2Cl2. The initial mixture is additionally stirred over night at room temperature, then sucked off and the filtrate is subjected to rotation processing. The remainder of 17.9 G. of Crude product is twice recrystallized from isopropanol. Yield 2.5 g (11,66% of theory.). Tpl.71,6aboutC.

P R I m e R 16. Obtaining the ethyl ester of N-(3-nitrobutyl)-cysteine.

I stage.

Saponification of the ethyl ester of 3-hydrogenases acid.

of 13.2 g (0.1 mol) of ethyl ester of 3-hydrogenases acid (Aldrich) is stirred 4.0 g (0, the scientists, the solution is acidified with 10 ml conc. HCl and twice extracted with him 100 ml of ethyl acetate. Then the solution is subjected to rotation processing, and receive a flowable oil.

Output 8,81 g (Teoret. 10.4 g of 3-hydroxybutiric acid.

II stage.

Nitration of 3-hydroxybutiric acid.

8,81 g (0.08 mol) of 3-hydroxybutiric acid and 50 mg of urea are dissolved in 50 ml of acetic acid at a temperature of 5aboutC. first added dropwise 6,27 ml (0.15 mol) of HNO3and then upon cooling, added dropwise 14,17 ml (0.15 mol) Ac2O. the Reaction mixture was stirred over night. When the processing to the resulting solution was added 200 ml of ice water and extracted it with ethyl acetate. The organic phase is extracted with NaHCO3. Phase NaHCO3acidified with conc. HCl and extracted with ethyl acetate. Then the solution is subjected to rotation processing, and receive a flowable oil. Yield 9.4 g (Teoret. 11.9 g of 3-nitrotoluenes acid.

Stage III. Obtaining the ethyl ester of N-(3-nitrobutyl)-cysteine.

16 g (0.11 mol) of 3-nitrotoluenes acid dissolved in 100 ml of dichloromethane. When applying N2and the temperature of the 15aboutWith added slowly 17.9 g (0.12 mol) of ethyl ester of cysteine. Then slowly GRE 15aboutAnd the filing of N2. After completion of the reaction afterwards received dicyclohexylphosphino and washed with a solution of 150 ml of 0.1 G. of HCl. Then the solution is subjected to rotation processing.

Do the cleaning substances by preparative chromatography on a column and by recrystallization from ethanol/n-hexane.

Output to 6.88 g (Teoret. 30,83 g).

T.PL.77,8aboutC.

P R I m e R 17. Obtaining the ethyl ester of N-(3-nitrobutyl)-methionine.

6,35 g (0,043 mol) 3-nitrotoluenes acid, 7.47 g (0,043 mol) of ethyl ester of methionine and volume of a spatula of dimethylaminopyridine (DMA) under stirring and cooling to 10aboutWith dissolved in 100 ml of dichloromethane. 10,31 g (0.05 mol) DCC dissolved in 80 ml of CH2Cl2while feeding nitrogen dropwise added slowly. After completion of the reaction solution is sucked off and washed it NaHCO3and then HCl. The solution is subjected to rotation processing, and get the oil.

Do the cleaning substances by preparative chromatography on a column and by recrystallization in cold weather.

The output of 1.95 g (Teoret. 12,05 g) ethyl ester of N-(3-nitrobutyl)-methionine in the form of colorless is aluchemie methyl ester NITROPHENOL acid.

25,0 g (to 0.19 mol) of methyl ether hydroxypivalic acid and 0.12 g of urea are dissolved at room temperature in 250 ml of CH2Cl2and cooled with stirring to 5aboutC. To this mixture with stirring is added dropwise to 23.8 g (0.38 mol) of 100% HNO3so that the temperature did not exceed 10aboutC. Then cooled to 5aboutC and stirring is added dropwise and 38.6 g (0.38 mol) of acetic anhydride so that the temperature did not exceed 10aboutC. for 15 min while cooling mix the mixture in an ice bath. Then slowly heat it to room temperature and during the night continue stirring at room temperature. The initial mixture slowly served while stirring in 500 ml of ice water. Separate CH2CL2-phase and once washed her 100 ml of dist. H2O, once with 100 ml saturated water solution of NaHCO3and again with 100 ml of dist. H2O. the Extract of CH2Cl2then concentrate in rotavapor when the bath temperature at Max. size 40aboutWith the vacuum generated by water-jet pump, until dry. Light yellow, oily residue is distilled in vacuum pump oil at a temperature of Vann stage. Getting NITROPHENOL acid.

14.0 g (0,350 mol) of NaOH dissolved in diest. H2O and cooled to about 10aboutC. To this mixture with stirring, a solution consisting of 31.0 g (0,175 mol) methyl ester NITROPHENOL acid in 250 ml of methanol, and the reaction mixture turns yellow and the temperature rises to 25aboutC. After stirring for 90 min, the initial mixture is neutralized 29.5 ml (0.35 mol) of 37% HCl. The methanol on a rotary evaporator (rotavapor) is completely distilled off. The water phase is twice extracted with 200 ml of methylene chloride. The combined methylene chloride extracts once washed with 50 ml of dist. H2O and methylenchloride phase concentrated on a rotary evaporator (rotavapor) until dry. Colorless, oily residue is dissolved in 100 ml of ethyl acetate and again concentrate on a rotary evaporator (rotavapor) until dry, and get a solid, white balance, which remove any traces of solvent in vacuum pump oil (of 0.4 Torr) at a temperature of bath 40aboutC and for 15 min on a rotary evaporator (rotavapor). Solid and white balance - 25,44 g (89,1% of Teoret.), dissolved in 100 ml of boiling n-hexane and to it dobavljalo, the product crystallizes. Product support for 72 h at a temperature of 0aboutWith, the crystals are sucked off and after double washing with 10 ml n-hexane cooled it at about 2 Torr and at room temperature in a vacuum drying to constant weight.

SoPL. 54,2aboutC.

Output 23,66 = 82,9% of theory.

Stage III. Obtaining the ethyl ester of N-(3-nitratomethyl)-cysteine.

10.7 g (71,7 mmol) ethyl ester of L-cysteine dissolved with stirring in nitrogen atmosphere in 200 ml of methylene chloride at room temperature. To the mixture of 11.4 g (70,0 mmol) of crystalline nitrosophenol acid, which is dissolved with stirring at room temperature. At room temperature and for 15 minutes, to this mixture are added dropwise with stirring and in a nitrogen atmosphere a solution consisting of 14.8 g (71,7 mmol) of N,N-dicyclohexylamine (DCC) in 50 ml of methylene chloride, and the temperature rises to 35aboutC. After further stirring the precipitated white dicyclohexylamine. The original mixture is cooled to room temperature and stirred under nitrogen atmosphere overnight. Then dicyclohexylphosphino sucked off through the porous Stekla is washed with 100 ml of 1N. HCl and twice with 100 ml of dist. H2O (in nitrogen atmosphere). Then they concentrate in rotavapor when the bath temperature is approximately 40aboutAnd in the vacuum produced by water-jet pump, from the first 550 mbar to about 20 mbar. Get a light brown oil.

The output of 21.2 g = 102.9% of theory.

The substance is purified by crystallization from ethanol/hexane at low temperature conditions.

Output 13,42 g = 65,1% of Torit. ethyl ester of N-(3-NITROPHENOL)-cysteine as a pale pink oil.

IV stage. Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-acetylcysteine.

To 10.3 g (35,0 mmol) of ethyl ester of N-(3-NITROPHENOL)-cysteine dissolved in 70 ml of dichloromethane, cooled conditions with paramasivan added dropwise a solution consisting of 4.3 g (42.0 mmol) of acetic anhydride in 10 ml of dichloromethane. Then added dropwise with stirring and cooled conditions, the solution consisting of 5.0 g (49,0 mmol) of triethylamine in 20 ml dichloromethane. After the initial reaction mixture is washed with 1N. HCl, 10% aqueous solution of sodium bicarbonate and water. The dichloromethane extract was concentrated on a rotary evaporator (rotavapor) until dry. In chilled conditions and by adding seed crystals of 7.8 g of crystalline product (66,3% of theory.).

T.PL.< 5aboutC.

Stage IV (Option 1). Obtaining the ethyl ester of N-3(Mitropoulos)-S-butyrolactone.

When using 6.7 g (42.0 mmol) of butyric acid anhydride instead of acetic anhydride (see stage 4) under the same reaction conditions and processing gain of 13.0 g of light yellow oily product, which as described above (stage IV) by crystallization under low temperature conditions obtain 9.7 g of crystalline product (76.2% of theory.).

Sopl.< 5aboutC.

Stage IV (Option 2). Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-devalorization.

When using 7,8 g (42.0 mmol) of anhydride pavlinovoi acid instead of acetic anhydride (see phase IV) under the same reaction conditions and processing obtain 14.1 g of a light yellow oily product, which as described above (stage IV) by crystallization under low temperature conditions get of 10.5 g of crystalline product (79,5% of theory.).

SoPL. 45aboutC.

Stage IV (Option 3). Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-cysteine-S-carbonate.

When using 4.3 g (42.0 mmol) of ethyl ether PI is the processing gain of 11.5 g of light yellow oily product, from which as described above (stage IV) by crystallization under low temperature conditions obtain 9.5 g of crystalline product (= 74,1% of theory.).

Sopl.36aboutC.

P R I m e R 19. Obtaining the ethyl ester of N-(3-NITROPHENOL)-methionine.

Dissolve with stirring and at room temperature of 12.4 g (70,0 mmol) ethyl ester of L-methionine in a nitrogen atmosphere in a 250 ml of methylene chloride. Add 11.4 g (70,0 mmol) of crystalline NITROPHENOL acid, which is dissolved with stirring at room temperature. To this mixture, with stirring and in a nitrogen atmosphere are added dropwise 14.8 g (71,7 mmol) N-N-dicyclohexylamine (DCC) in 50 ml of methylene chloride at room temperature and for 15 min, and the temperature rises to 35aboutC. After further stirring the precipitated white urea DC. The original mixture is cooled to room temperature and stirred overnight under nitrogen atmosphere. DCC-urea then sucked off through a porous glass filter and once washed with 50 ml of CH2Cl2. The combined solution of methylene chloride once washed with 100 ml of 1N. HCl and twice with 100 ml of dist. H2O (in nitrogen atmosphere), and then their concen using a water-jet pump, 550 mbar to 20 mbar. Get a light yellow oil.

Output 24,9 g = 110.3 per cent from Teoret. the crude ethyl ester of N-(3-NITROPHENOL)-L-methionine.

The crude product is purified by preparative chromatography on a column.

Output 17,6 g = 78,0% of theory. ethyl ester of N-(3-NITROPHENOL)-methionine in the form of a colorless oil.

P R I m e R 20. Obtaining N-(12-nitrothiazol)-S-acetylcysteine.

I stage. Obtain 12-nitrotoluene acid.

Dissolve to 54.1 g (0,250 mol) 12-hydroxylamino acid and 0.3 g of urea at low heat in a 1.3 l CHCl3and cooling the mixture with stirring to 20aboutFrom 23.6 g (0,375 mol) HNO3(100%) with stirring slowly added dropwise, and the temperature rises to 27aboutC. the mixture is Then cooled to 20aboutWith and, stirring, added dropwise to 38.3 g (0,375 mol) of acetic anhydride while cooling, and keep the temperature at least 25aboutC. during the night stirred the mixture at room temperature. Then washed 5 times with 0.5 l of SW. H2O. After drying over Na2SO4and clarification of powdered activated carbon concentrate CHCl3phase on a rotary evaporator (ro the state. Oily residue of 60.8 g dissolved in 500 ml of boiling n-hexane and after cooling to room temperature, leave it over night in the refrigerator at a temperature of 0aboutC. Bicrystalline the product is sucked off and washed with twice 50 ml of n-hexane. Then dried the product in a vacuum dryer at room temperature and about 2 Torr to constant weight.

SoPL. 29aboutC.

Output 39,4 g = 60.3% from Teoret.

II stage. Getting chloride 12-nitrotoluene acid.

Dissolve 2,61 g (10 mmol) nitrotoluenes acid in 50 ml of methylene chloride with stirring at room temperature are added dropwise 4.44 g (35 mmol) of oxalicacid in 50 ml of methylene chloride. The mixture overnight mix. The product is then concentrated in a rotary evaporator until dry.

Output 3 g = 93,2% of theory.

Stage III. Obtaining N-(12-nitrotoluene)-cysteine.

In the atmosphere of nitrogen is added with stirring the 6.06 g (50 mmol) of L-cysteine in 300 ml D. the ceiling of 5.60 g (20 mmol) of the chloride 12-nitrotoluene acid in 50 ml of dichloromethane is added dropwise. Due to the fact that you do not receive a clear solution, it is heated up to 60aboutC. Then add 100 UB>2
O and extracted four times with 200 ml of ethyl acetate. The organic phase is dried over Na2SO4and then her focus. The remainder is added to 100 ml of ether and placed in a refrigerator for bicrystalline during the night and at a temperature of 0aboutC. Obtain white crystals.

Sopl.74-75aboutC.

Yield 4.1 g of N-(12-nitrotoluene)-cysteine.

IV stage. Obtaining N-(12-nitrotoluene)-S-acetylcysteine.

In the atmosphere of nitrogen to prepare 1,82 g (5 mmol) of N-(12-nitrotoluene)-cysteine in 20 ml of ethyl acetate. Then cool the mixture to 0aboutC and added dropwise 2.5 ml of acetic anhydride. At a temperature of -5aboutWith dropwise added slowly 1.52 g (15 mmol) of triethylamine, dissolved in 5 ml of ethyl acetate. The reaction solution is washed with water and concentrated until dry.

Sopl.at room temperature butter.

Yield: 2 g = 98,4% of theory.

P R I m e R 21. Obtaining the ethyl ester of N-(12-nitrotoluene)-cysteine.

Dissolve 4 g (26.8 mmol) of ethyl ester of cysteine in 50 ml of methylene chloride and stirring is added dropwise 2.8 g (10 mmol) of the chloride 12-nitrotoluene acid, dissolved in 50 ml of m is the solvent on a rotary evaporator (rotavapor). Oily residue (6 g) dissolved in 100 ml of ether and leave in the refrigerator overnight at a temperature of 0aboutC. the Precipitated product is sucked off.

Sopl.59-60aboutC.

Yield 1.6 g = 40.0% from Teoret.

P R I m e R 22. Obtaining the ethyl ester of N-(2-nitropropionic)-cysteine.

I stage. Obtaining the ethyl ester nitromalonic acid.

33 g (0.28 mol) of ethyl ester of lactic acid is dissolved in 300 ml of dichloromethane. After adding 100 mg of urea at a temperature of 5-10aboutWith added dropwise to 22.5 ml of 0.56 mol) of 100% nitric acid. Cool the solution to a temperature of 0aboutC. and Then added dropwise to 52.8 ml of 0.56 mol) of acetanhydride so that the temperature did not exceed 5aboutC. Leave the solution overnight at room temperature, and then washed his 250 ml of water. The organic phase is separated and dried over sodium sulfate. After filtering distilled dichloromethane. The obtained oily residue is treated by distillation.

Output 30,34 g = 66,4% of theory.

So Kip. 34about(Of 0.25 Torr).

II stage. Getting nitromalonic acid.

30 g (0.18 mol) of ethyl ether nitromalonic acid rastgele for 19 PM The solution is concentrated to a volume of approximately 50 ml and then diluted it with 300 ml of water. The pH value was adjusted to 7-8 by addition of sodium bicarbonate. Untransformed ether is separated by extraction with dichloromethane. The aqueous phase is brought to pH 1 conc. hydrochloric acid and extracted three times its 150 ml of ethyl acetate. Combine the extracts and dry over sodium sulfate. After filtration, the ethyl acetate is completely removed on a rotary evaporator.

Yield 14.6 g of colorless oil = 59.2% of theory.

Stage III. Obtaining the ethyl ester of N-(2-nitropropionic)-cysteine.

In the atmosphere of nitrogen dissolve 17 g (0.13 mol) nitromalonic acid and 18.9 g (0.13 mol) of ethyl ester of cysteine at a temperature of 10-15aboutWith 200 ml of dichloromethane. At a temperature of 15-20aboutWith added dropwise a solution consisting of 28,6 (0.14 mol) of N,N-dicyclohexylcarbodiimide and 75 ml of dichloromethane. After 1 h, filtered off the precipitated N,N-dicyclohexylamine and additionally washed her 75 ml dichloromethane. The filtrate is extracted twice with 50 ml of 0.1 N. hydrochloric acid. The organic phase is completely concentrated on a rotary evaporator. The crystalline crude product (22,4 g) is recrystallized from 100 ml of ethanol/n-hexane (1:1).these radicals may serve as the following:

A) Deputy

1) R1= hydrogen, C1-C4-alkyl, amino, acylamino, hydroxy, acyloxy.

As an example, for R1= acylamino the applicant shall indicate on the receipt of ethyl ester of N-(nitrate-N'-acetyl-D,L-seryl)-cysteine.

Connection O-acyl and oxycoedone can be obtained in the same way.

R1= acylamino

Obtaining the ethyl ester of N-(nitrate-N'-acetyl-D,L-seryl)-cysteine.

I. Stage. Getting nitrate-N-acetyl-D,L-serine.

of 11.8 g (0.08 mol) of N-acetyl-D,L-serine are dissolved in 50 ml of acetic acid. At a temperature of 5-10aboutWith added dropwise 6,27 ml (0.15 mol) of HNO3and then 14,17 ml (0.15 mol) AU2O. the Reaction mixture was stirred over night. To the resulting solution was added 200 ml of ice water and extracted it with ethyl acetate. The organic phase is extracted with a solution of NaHCO3. Acidified phase NaHCO3with conc. HCl and extracted it with ethyl acetate. The extract obtained is subjected to rotation processing, and get a plastic butter.

Physical parameters no, because the product is an intermediate product.

Output 11,98 g = 72% from Teoret.

II stage. Getting ethyl everaere nitrogen and with stirring, dissolved in 200 ml of methylene chloride at room temperature. Add 11,98 g nitrate-N-acetyl-D,L-serine, which is dissolved with stirring at room temperature. At room temperature and for 15 minutes, to this mixture are added dropwise with stirring and in a nitrogen atmosphere a solution of 14.8 g (71,7 mmol) N,N-dicyclohexylamine (D) in 50 ml of methylene chloride, and the temperature rises to 35aboutC. After further stirring the precipitated white dicyclohexylamine. The original mixture is cooled to room temperature and stirred under nitrogen atmosphere overnight. Then dicyclohexylphosphino sucked off through a porous glass filter and once it is washed with 50 ml of CH2Cl2. United methylenchloride solutions once washed with 100 ml of 1 N. HCl and twice with 100 ml of dist. H2O (in nitrogen atmosphere). Then they concentrate in rotavapor when the bath temperature is approximately 40aboutAnd in the vacuum produced by water-jet pump, from the first 550 mbar to about 20 mbar. Get a light brown oil. According SIH (MPLC) receive 4.26 deaths g butter++= 21% from Teoret. (VGH: 98,3%).

++MS: m+323 and (m+1)+324 received.

Fragmentation confirms the structure.

2) R2= hydrogen or C1-C4-SUB>1-C4-alkyl, C1-C6-alkanoyl tio1-C4-alkyl, in particular alkylen-S-pivalate, alkylen-S-butyrate, alkylen-S-propionate, alkylen-S-acetate, alkylene-S-hexanoate, alkylen-S-octanoate, alkylen-S-benzoate, alkylen-S-actinometry ether, alkylene-thiocarbonate, alkylen-THIOCARBAMATE, alkylen-dionissia alkyl, lower alkyl-dionissia alkalicarbonate acid and/or their esters and/or amides.

The following examples confirm vysheukazannye values of R5:

R5= methylene-S-hexanol, methylene-S-octanol, methylene-S-benzoyl, methylene-S-actinometry ether, methylthiocarbamate (CH2-S-CO2-R).

Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-hexanolactone.

Dissolve 20 g (0.07 mol) of ethyl ester of N-(3-NITROPHENOL)-cysteine in 200 ml of dichloromethane and added 11.2 ml (0.08 mol) of hexanolactone. Then at a temperature of 5-10aboutWith added dropwise 11 ml (0.08 mol) of triethylamine. The initial mixture is additionally stirred for 15 h and then washed it successively with 5% HCl, conc. a solution of NaHCO3and water. The dichloromethane phase is concentrated. According SIH (MPLC) get of 8.28 g butter++(31,25% of theory. , BIH: 97,3%);++MS: m+LASS="ptx2">

Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-(carbetimer)-cyst - in.

Dissolve to 14.4 g (0.061 mol) of ethyl ester carbethoxymethylthio and 11,05 g (0,068 mol) 3-NITROPHENOL acid in 150 ml of dichloromethane. At a temperature of 15-25aboutWith added dropwise 14 g (0,068 mol) dicyclohexylcarbodiimide (DCC), dissolved in 50 ml of dichloromethane. The initial mixture is additionally stirred at room temperature for 15 hours and Then filtered urea and the filtrate concentrated. After processing by chromatography on a column of gain of 8.5 g butter++= 36.6% from Teoret.

++MS: m+380 (m+1)+381 see. Fragmentation confirms the structure.

R5= CH2-S-CONH2.

Obtaining the ethyl ester of N-(3-NITROPHENOL)-S-carbamoylation.

0.05 mol of ethyl ether carbamoylation and 3-NITROPHENOL acid dissolved in 100 ml of dichloromethane. At room temperature, added dropwise to 0.5 mol of dicyclohexylcarbodiimide dissolved in 50 ml of dichloromethane. After processing by chromatography on a column of get a product with the release of 24%; Sopl.58aboutC.

6) R5and R thiolactone on NITROPHENOL acid and 4.7 g (0.03 mol) of homocysteinaemia x HCl in 50 ml of THF. After adding 2.4 g of pyridine at a temperature of 5-10aboutWith added dropwise a solution of 6.2 g of dicyclohexylcarbodiimide (DCC) in 20 ml of THF. The product is processed by chromatography on a column and then it will recrystallized from hexane/EtOH.

The yield of 0.51 g = 6.5% of theory. T.PL. 88,6aboutC.

Elemental analysis showed: 41,43% (41,21%); 5,44% N (5,38%); 30,1% (30,5%).

7) R = hydroxy, C1-C4-alkoxy, amino, C1-C4-alkylamino, di-C1-C4-alkylamino, low-alkylamines-alkoxy, Denizli alkylamino-lower alkoxy, acyloxy-lower alkoxy, arlinski alkoxy, substituted arlinski alkoxy, where the Deputy means methyl, halogen or methoxy, or amino acid residues selected from the group of cysteine, methionine or homocysteine attached peptide bond.

Relevant examples can serve as an extension of example 16, stage III.

R = Denizli-alkyl-eminently alkoxy

acyloxy-lower alkoxy

arlinski alkoxy

Addition to example 16, stage III.

When using the corresponding esters of cysteine in the same way receive the ethyl ester of N-(3-nitrobutyl)-containtyrosine (oil is l)-cysteine (Sopl.33aboutC) and ethyl ester of glycolic acid N-(3-nitrobutyl)-cysteinyl (Sopl.48aboutC).

8). An example illustrating R = peptide.

Dissolve 4 g (10 mmol) N-12-nitrotoluene-S-acetylcysteine and 10 mmol of the peptide in 80 ml of dichloromethane. At room temperature is added dropwise a solution of 2.06 g (10 mmol) dicyclohexylcarbodiimide (DCC) in 20 ml of dichloromethane. The initial mixture is additionally stirred at room temperature for 15 hours Urea DCC will be filtered and dichloromethane completely removed. The remainder is processed using SSH.

Output a 3.87, Sopl.68,4about(Diisopropyl ether).

Joining peptide N-end-to-end N-12-nitrotoluene-S-acetylcysteine is similar to that of the synthesis of the below Pentapeptide - Tyr-Pro-Phe-Pro-Gly-methyl ether.

9) Synthesis of Pentapeptide having the sequence: Tyr-Pro-Phe-Pro-Gly-methyl ether.

(a) the Plan synthesis.

Amine component interacts at a temperature of -15aboutC or lower in dimethylformamide (DMF) with 0.5 molar excess mixed anhydride isobutylketone acid, Z-amino acids (and Z is a protective group and mean residue> who or below for 10-15 min using 6% of the excess of the derived Z-amino acids and N-methylmorpholine through isobutyl ether of Harborview acid. Excess mixed anhydride and then destroy. At a temperature of 0aboutWith water and a saturated solution KHCO3bring the pH of the reaction product up to the value of 8 and stirred for 30 minutes

Peptides extracted with ethyl acetate. To remove the potassium salt of Z-amino acid washed with a mixture of ethyl acetate and peptide three times with sodium chloride/water and three times with water and then evaporated. Thus obtained peptide, which still has the protective group Z, hydronaut in methanol. When this type of 100-500 mg of Pd/active charcoal catalyst in mmol of the peptide. Dissociation of CO2control solution of BA(OH)2. The catalyst is filtered (filter paper No. 595 of the firm Schleicher &Schull), thoroughly washed with water and the filtrate vparivayut on a rotary evaporator (type the rotavapor, Buchi company). The desired released peptide is in the balance.

b) Synthesis of Pentapeptide

Tyr-Pro-Phe-Pro-Gly-methyl ether.

Stage I.

a) Obtaining a mixed anhydride (Z-Phe-Pro-mixed anhydride).

640 mg (1.6 mmol = 6% excess) dippy) is subjected to interaction in 20 ml of dimethylformamide (DMF) and 200 μl (1.5 mmol) isobutyl ether of Harborview acid at a temperature of -15aboutC and for 15 min after addition of 170 μl (1.6 mmol) of N-methylmorpholine.

b) preparation of the amine component.

125,6 mg (1.0 mmol) of getinputfiledirectory dissolved in 20 ml of DMF with the addition of 100 μl (1 mmol) N-methylmorpholine at a temperature of -15aboutC.

Stage II.

The reaction of the mixed anhydride of stage 1A with aminoven component 1B. Z-Phe-Pro-mixed anhydride is subjected to interaction with glycymeris ether in 40 ml DMF at -15aboutWith and within 4 hours

z-Phe-Pro-mixed anhydride+Gly-METAVIR -15aboutWith- - - - -> z-Phe-Pro-METAVIR 4 hours

Before processing destroy 50% excess mixed anhydride. At a temperature of 0aboutWith water, a saturated solution of knso3bring the pH of the reaction product up to the value of 8 and stirred for 30 min at a temperature of 0aboutC. and Then extracted peptide 50-100 ml of ethyl acetate (EtAc). A mixture of ethyl acetate and the peptide was washed with saturated aqueous solution of sodium chloride. After the final rinse water is evaporated phase of ethyl acetate.

Stage III. Cleavage of the protective groups by hydrogenation.

Dissolve the peptide in 30 ml of methanol and add 100 mg of palladium on assistaut at a temperature of 25-30aboutC. the Hydrogenation is completed, when no longer exempt CO2i.e. after controlling in an aqueous solution of barium hydroxide is no longer a precipitate. The solution is filtered, washed with water and subjected to its rotation processing on a rotary evaporator. The remaining intermediate product is used as amine component at stage IV.

Stage IV.

a) Obtaining a mixed anhydride (Z-Pro-mixed. anhydride).

Dissolve 374 mg (1.5 mmol) of Z-L-Proline in 15 ml of DMF adding 170 μl (1.5 mmol) N-methylmorpholine and react a solution of 180 μl (1.4 mmol) of isobutyl ether of Harborview acid at a temperature of -15aboutC for 15 minutes

b) Reaction of the mixed anhydride of the stage IV with aminoven component of stage III. Z-L-Pro-anhydride + Phe-Pro-Gly-metaphor (in

-15aboutWith 15 ml of DMF) - - - - - - - - -> Z-Pro-Phe--Pro-Gly-metaphor.

4 h the Destruction of excess mixed anhydride, extraction, hydrogenation is carried out in accordance with the above.

The final product IV is aminoven component for stage V. the Control on the composition of amino acids after hydrolysis showed that the peptide has the correct molar ratio of amino acids.

Study the di-Z-L-tyrosine in 15 ml of DMF adding 165 μl (1.4 mmol) of N-methylmorpholine and the solution is subjected to interaction with 175 μl (1.3 mmol) of isobutyl ether of Harborview acid at a temperature of -15aboutWith and within 15 minutes

b) Reaction of the mixed anhydride stage VA with the final product stage IV. The final product stage IV dissolved in 15 ml of DMF and subjected to interaction with the mixed anhydride stage 5A at a temperature of -15aboutWith and within 4 hours

The destruction of excess mixed anhydride, extraction and hydrogenation is carried out in accordance with the above.

The amino acids analysis, carried out after hydrolysis in the acidic environment, showed the correct molar ratio of amino acids in accordance with Pentapeptide Tyr-Pro-Phe-Pro-Gly-methyl ether.

10) m, n and o are 0, and/or 1-10.

b) Stereochemical forms of the amino acids used.

Used amino acids, such as cysteine, were obtained from the company Fluka. They got a D-amino acids and L-amino acids.

Principally used L-amino acids. Racemic D/L-amino acids or D-amino acids can be used in the same way.

In addition to example 20, step III, the following option can be entered.

Obtaining N-(12-nitrotoluene)-D-cysteine.

If we replace L-cysteine 6,06 g D-cysteine (Fluka), at the same slowianie ethyl ester of N-(3-NITROPHENOL)-methionine.

12.4 g (70,0 mmol) ethyl ester of L-methionine was dissolved with stirring in nitrogen atmosphere and at room temperature in 250 ml of methylene chloride. Add 11.4 g (70,0 mmol) of crystalline NITROPHENOL acid and dissolved with stirring at room temperature. Stirring, to this mixture are added dropwise in a nitrogen atmosphere a solution of 14.8 g (71,7 mmol) N,N-dicyclohexylamine in 50 ml of methylene chloride for about 15 min and at room temperature, and the temperature rises to 35aboutC. After additional stirring the precipitated white dicyclohexylamine. The original mixture is cooled to room temperature and stirred overnight under nitrogen atmosphere. N,N-dicyclohexylamine then sucked off through a porous glass filter and once it is washed with 50 ml of CH2CH2. The combined solution of methylene chloride once washed with 100 ml of 1 N. HCl and twice with 100 ml of dist. H2About (in nitrogen atmosphere) and then concentrated in rotavapor at a temperature of bath 40aboutC and under vacuum, obtained by water-jet pump, 550 mbar to 20 mbar. Get a light yellow oil.

Output 24,9 g = 110.3 per cent from Teoret. the crude ethyl ester of N-(3-narutopedia is 0% of theory. ethyl ester of N-(3-NITROPHENOL)-methionine in the form of a colorless oil.

In this case, for inclusion in the scope of protection1-C4-alkylthio-C1-C4-alkyl as the value of the radical R5specifies the physical parameter is the refractive index, which is set to 1,4897 at a temperature of 25aboutC.

An example illustrating the formation of salts.

Getting the Na-salt of N-(12-nitrotoluene)-S-devalorization.

446 mg (1 mmol) of N-(12-nitrotoluene)-S-revalorization dissolved in 50 ml of ethyl acetate. In an atmosphere of nitrogen was slowly added dropwise a solution consisting of 40 mg (1 mmol) of NaOH and 5 ml of ethanol. The precipitated product is sucked off and dried in vacuum.

The output of 480 mg T.PL.118-125aboutC (Z).

Getting triethanolamine salt of N-(3-NITROPHENOL)-S-devalorization.

of 1.75 g (5 mmol) of N-(3-NITROPHENOL)-S-revalorization dissolved in 20 ml of dichloromethane. In nitrogen atmosphere and at room temperature is added dropwise to 750 mg (5 mmol) of triethanolamine dissolved in 10 ml of dichloromethane. Dichloromethane completely removed.

Output of 2.5 red oil++which hardens at a temperature of 5-7aboutC.

With regard to toxicity, the applicant notes that the compounds according to the invention have a low toxicity. Connection ethyl ester of N-NITROPHENOL-S-(N-acetylenyl)-L-cysteine and ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine were tested for toxicity as representatives of the class of substances. According to the test results non-toxic dose of 200 mg of the compounds according to the invention per 1 kg of body weight. In accordance with the strict guidelines for toxicity of drugs every day to 160 mg of the compounds according to the invention can be taken over long periods of time without side effects.

Pharmacological data for the selected compounds according to the invention:

(a) Pharmacological in vivo model (waking the dog)

Hemodynamic actions ethyl ester N-NITROPHENOL-S-(N-acetylethyl)-L-cysteine and ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine after intravenous and oral administration experience by direct comparison with the ethyl ester of N-NITROPHENOL-S-pivaloyl-L-cysteine and classic organic nitrate - isosorbide-5-Mononitrate (IS-5-N) in the awake trained dogs genus is RA, introduced through V,hugularis. Arterial system describe the systolic, mean and diastolic blood pressure and heart rate, and venous system of Central venous and pulmonary arterial pressure. At the same time determine minute volume of blood by thermodilution and calculate the total peripheral resistance and elasticity of the arterial vascular system.

Both substances, namely ethyl ester of N-NITROPHENOL-S-(N-acetylethyl)-L-cysteine and ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine are in intravenous and oral introduction, the range of actions typical of nitrate, however, they generally have a considerably strong effect than the comparative substance, i.e., the ethyl ester of N-NITROPHENOL-S-pivaloyl-L-cysteine and is-5-n In the table.1 shows a representative comparison of the levels of action after administration of equimolar doses of the respective substances (1.7 mcmol/kg intravenously) in the awake dog (n = 5):

b) Pharmacological in vitro model (isolated aortic rings):

The action of the above nitrates also have in vitro with respect to their effect relaxation of blood vessels. As the models use an isolated Aorta thoracalis adventitious tissue, in the solution of Krebs-Henseleit temperature 37aboutC and in an atmosphere of oxygen, vascular rings with pre-loading at the rate of 2 g pre-cut 2x10-7M phenylephrine. After reaching a stable level of reduction perform cumulative addition of the corresponding nitrates in the tub, and isometrically record the development of the power to obtain the curve of concentration and action. In this model, compounds, namely, ethyl ester N-NITROPHENOL-S-(N-acetylethyl)-L-cysteine and ethyl ester of N-NITROPHENOL-S-(N-acetylsalicyl)-L-cysteine are also showing a greater effect than nitrates, i.e., the ethyl ester of N-NITROPHENOL-S-pivaloyl-L-cysteine and is-5. the Following is a comparison of the established concentrations (EC50), which cause premaxillae the reduced relaxation of aortic rings:

The above pharmacological tests have shown that the compounds according to the invention, thanks to the connecting component, i.e., N-acetylaminocinnamic, in particular N-acetylglycine or N-acetylamino, respectively, have significantly high effect of nitrate than the comparative substance - ethyl ester N-NITROPHENOL-S-pivaloyl-L-cysteine. It stands for tera the purpose of these new substances as products of hydrolysis only occur amino acids - glycine and alanine, which are already present in micromolar concentrations in the plasma.

Derivatives nitroalkanes acids of General formula

O2N-O-CH2-(CH2)m--(CH2)n-(CH2)o--R

where R is lower alkoxy or attached peptide bond Pentapeptide Tyr - Pro - Phe - Pro - Gly(OMe);

R1is hydrogen, C1-C6- alkyl, acylamino;

R2is hydrogen, lower alkyl;

R3is hydrogen,

R4is hydrogen, lower alkyl;

R5- (CH2)kSX, where X is hydrogen, lower alkyl or X - COR11,

where R11is hydrogen, lower alkyl or a residue of N-acylaminoalkyl;

m = 0.1 to 10;

n and o = 0 to 2,

or their pharmaceutically acceptable salts.

 

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The invention relates to obstetrics and can be used in hospitals when you stop hypotonic uterine bleeding in childbirth, at the time of caesarean section and postpartum period

FIELD: pharmaceutical chemistry.

SUBSTANCE: invention relates to (i) essentially crystalline melagatran in the form of hydrate, which is characterized by x-ray diffraction pattern on powder having crystalline peaks with following d values: 21.1, 10.5, 7.6, 7,0, 6.7, 6.4, 6.2, 5.7, 5.4, 5.3, 5.22, 5,19, 5.07, 4.90, 4.75, 4,68, 4.35, 4.19, 4.00, 3.94, 3.85, 3.81, 3.73, 3.70, 3.63, 3.52, 3.39, 3.27, 3,23, 3.12, 3.09, 3.06, 2.75, 2.38, and 2.35 Å and/or water content 4.3%; and (ii) essentially crystalline melagatran in the form of anhydrate, which is characterized by x-ray diffraction pattern on powder having crystalline peaks with following d values: 17.8, 8.9, 8.1, 7.5, 6.9, 6.3, 5.9, 5.6, 5.5, 5.4, 5.3, 5.2, 5.0, 4.71, 4.43, 4.38, 4.33, 4.14, 4.12, 4.05, 3.91, 3.73, 3.61, 3.58, 3.56, 3.47, 3.40, 3.36, 3,28, 3.24, 3.17, 3.09, 3.01, 2.96, 2.83, 2.54, 2.49, 2.41, 2.38, and 2.35 Å. Invention also relates to a method for preparation of indicated form, a method for interconversion of anhydrite form, to use of indicated compounds as pharmaceutical agent, and to preparation of drugs. Pharmaceutical preparation is suitable for treatment of condition, in case of which inhibition of thrombin is needed or desirable. Invention provides a method for treatment of such condition.

EFFECT: increased chemical stability and solid state stability as compared to amorphous forms of melagatran.

14 cl, 4 dwg, 3 tbl, 9 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to compounds of the formula (I):

wherein r = 1, 2 or 3; s = 0; t = 0; R1 is taken among group including R11-CO and R12-SO2- wherein R11 is taken among group including (C6-C14)-aryl, (C1-C8)-alkyloxy-group wherein all given group are unsubstituted or substituted with a single or some similar or different substitutes R40; R12 means (C6-C14)-aryl wherein indicated group is unsubstituted or substituted with a single or some similar or different substituted R40; R2 means R21(R22)CH-, R23-Het-(CH2)k-, R23(R24)N-(CH2)m-D-(CH2)n- or R25(R26)N-CO-(CH2)p-D-(CH2)q- wherein D means bivalent residue -C(R31)(R32)-, bivalent (C6-C14)-arylene residue or bivalent residue obtained from aromatic group Het comprising 5 or 6 atoms in cycle among them 1 or 2 are similar or different cyclic heteroatoms taken among group including nitrogen and sulfur atoms; numbers k, m, n, p and q = 0, 1, 2; R21 and R22 that are independent of one another can be similar or different and taken among group including hydrogen atom, (C1-C12)-alkyl, (C6-C14)-aryl and so on; R23 means hydrogen atom, R27-SO2- or R28-CO-; R24, R25 and R26 mean hydrogen atom; R27 is taken among group including (C1-C8)-alkyl, (C6-C14)-aryl and so on; R28 is taken among group including R27, (C1-C8)-alkyloxy-group; R31 and R32 mean hydrogen atom; R40 is taken among group including halogen atom, hydroxy-, (C1-C8)-alkyloxy-group, (C1-C8)-alkyl, (C6-C14)-aryl and so on; R91, R92, R93 and R96 means hydrogen atom; R95 means amidino-group; R97 means R99-(C1-C8)-alkyl; R99 is taken among group including hydroxycarbonyl- and (C1-C8)-alkyloxycarbonyl-; Het means saturated, partially unsaturated or aromatic monocyclic structure comprising from 3 to 6 atoms in cycle among them 1 or 2 are similar or different heteroatoms taken among group comprising nitrogen and sulfur atoms; in all its stereoisomeric forms and also their mixtures in any ratios, and its physiologically acceptable salts. Invention proposes a method for preparing compound of the formula (I). Also, invention proposes a pharmaceutical preparation eliciting inhibitory activity with respect to factor VIIA and containing at least one compound of the formula (I) and/or its physiologically acceptable salts and pharmaceutically acceptable carrier. Invention provides preparing compounds of the formula (I) eliciting power anti-thrombosis effect and useful for treatment and prophylaxis of thrombosis-embolic diseases.

EFFECT: valuable medicinal properties of compounds and composition.

10 cl, 70 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to compounds of the formula (I):

wherein r = 1, 2 or 3; s = 0; t = 0; R1 is taken among group including R11-CO and R12-SO2- wherein R11 is taken among group including (C6-C14)-aryl, (C1-C8)-alkyloxy-group wherein all given group are unsubstituted or substituted with a single or some similar or different substitutes R40; R12 means (C6-C14)-aryl wherein indicated group is unsubstituted or substituted with a single or some similar or different substituted R40; R2 means R21(R22)CH-, R23-Het-(CH2)k-, R23(R24)N-(CH2)m-D-(CH2)n- or R25(R26)N-CO-(CH2)p-D-(CH2)q- wherein D means bivalent residue -C(R31)(R32)-, bivalent (C6-C14)-arylene residue or bivalent residue obtained from aromatic group Het comprising 5 or 6 atoms in cycle among them 1 or 2 are similar or different cyclic heteroatoms taken among group including nitrogen and sulfur atoms; numbers k, m, n, p and q = 0, 1, 2; R21 and R22 that are independent of one another can be similar or different and taken among group including hydrogen atom, (C1-C12)-alkyl, (C6-C14)-aryl and so on; R23 means hydrogen atom, R27-SO2- or R28-CO-; R24, R25 and R26 mean hydrogen atom; R27 is taken among group including (C1-C8)-alkyl, (C6-C14)-aryl and so on; R28 is taken among group including R27, (C1-C8)-alkyloxy-group; R31 and R32 mean hydrogen atom; R40 is taken among group including halogen atom, hydroxy-, (C1-C8)-alkyloxy-group, (C1-C8)-alkyl, (C6-C14)-aryl and so on; R91, R92, R93 and R96 means hydrogen atom; R95 means amidino-group; R97 means R99-(C1-C8)-alkyl; R99 is taken among group including hydroxycarbonyl- and (C1-C8)-alkyloxycarbonyl-; Het means saturated, partially unsaturated or aromatic monocyclic structure comprising from 3 to 6 atoms in cycle among them 1 or 2 are similar or different heteroatoms taken among group comprising nitrogen and sulfur atoms; in all its stereoisomeric forms and also their mixtures in any ratios, and its physiologically acceptable salts. Invention proposes a method for preparing compound of the formula (I). Also, invention proposes a pharmaceutical preparation eliciting inhibitory activity with respect to factor VIIA and containing at least one compound of the formula (I) and/or its physiologically acceptable salts and pharmaceutically acceptable carrier. Invention provides preparing compounds of the formula (I) eliciting power anti-thrombosis effect and useful for treatment and prophylaxis of thrombosis-embolic diseases.

EFFECT: valuable medicinal properties of compounds and composition.

10 cl, 70 ex

FIELD: pharmaceutical chemistry.

SUBSTANCE: invention relates to method for production of acetylamidiniophenylalanylcyclohexylglycilpypidinioalanin amides of formula I , wherein X anions are physiologically acceptable anions, and analogous thereof. Said compounds are effective inhibitors of fibrillation factor Xa and are useful, for example, in prevention of thrombosis. Claimed method includes coupling of 2-[2-acetylamino-3-(4-amidinophenyl)-propionylamino]-2-cyclohexylacetic acid, obtained from 2-[2-acetylamino-3-(4-cyanophenyl)acryloylamino]-2-cyclohexylacetic acid by assimetric hydration and converting of cyano group to amidine, or salt thereof with 3-(2-amino-2-carbamoylethyl)-1-methylpyridinic acid or salt thereof. Also are disclosed starting materials and intermediated used in this method, process for production the same and acetyl-(S)-4-amidiniophenylalanyl-(S)- cyclohexylglycil-(S)-(1-methyl-3-pypidinio)alanin amide in form of ditosylate.

EFFECT: simplified method; increased commercial availability of compounds with applicable anion.

14 cl, 16 ex

FIELD: organic chemistry, medicine, pharmacology.

SUBSTANCE: invention relates to new inhibitors of thrombin of the formula (I)

,

method for their preparing, intermediate compounds used for their preparing of the formula (II)

and a pharmaceutical composition comprising compounds of the formula (I). Invention provides enhancing effectiveness in inhibition of thrombin.

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

23 cl, 61 ex

FIELD: chemistry of peptides, microbiology, biotechnology.

SUBSTANCE: L-alanyl-L-glutamine is prepared by incubation of a mixture containing a microorganism able to produce L-alanyl-L-glutamine from L-alanine ester and L-glutamine, L-alanine ester and L-glutamine, and isolation of the end product. Using the invention allows simplifying the process for preparing L-alanyl-L-glutamine. Invention can be used in pharmacy and food processing industry.

EFFECT: improved preparing method of dipeptide.

3 cl, 3 tbl, 1 ex

Peptide compounds // 2281955

FIELD: chemistry of peptides, medicine, pharmacy.

SUBSTANCE: invention relates to compound of the formula (I): wherein R1 represents benzofuranyl substituted with halogen atom or styryl substituted with halogen atom; R2 represents substituted hydroxyl substituted with mercapto-group or substituted sulfonyl, or its pharmaceutically acceptable salts. Compound of the formula (I) and its pharmaceutically acceptable salts possess the strong inhibitory effect on production of nitrogen oxide (NO) and can be useful in prophylaxis and/or treatment of NO-mediated diseases in humans and animals.

EFFECT: valuable medicinal and biochemical properties of compounds.

FIELD: pharmaceutical dosage forms.

SUBSTANCE: invention relates to compositions and methods for delivery of drugs. Composition according to invention represents powdered composite containing polymer, therapeutical agent, and complexing agent, said polymer containing one or more cyclodextrin fragments. Polymer interacts with complexing agent in a host/guest or guest/host manner resulting in formation of inclusion complex. Both polymer of composite and complexing agent may be used in a way to introduce functionality into therapeutical composition. Invention also discloses a method for preparing composition and to a method for delivering therapeutical agent.

EFFECT: widened choice of drug delivery methods.

26 cl, 31 dwg, 1 tbl, 65 ex

FIELD: organic chemistry, medicine, endocrinology, pharmacy.

SUBSTANCE: invention relates to novel compounds of the formula (I): or their pharmaceutically acceptable salts and composition containing thereof that possess activity with respect to release of growth hormone. In the formula (I) R1 represents hydrogen atom or (C1-C6)-alkyl; R2 represents hydrogen atom or (C1-C6)-alkyl; L represents compound of the formula: wherein R4 represents hydrogen atom or (C1-C6)-alkyl; p = 0 or 1; r = 1; q, s, t and u = 0, 1, 2, 3 or 4 being independently of one another; the sum q + r + s + t + u = 0, 1, 2, 3 or 4; R9, R10, R11 and R12 represent independently of one another hydrogen atom or (C1-C6)-alkyl; Q represents >N-R12 wherein R13 represents hydrogen atom or (C1-C6); or L represents compound of the formula: , wherein r = 0 or 1; q, s, t and u independently of one another = 0, 1, 2, 3 or 4; the sum q + r + s + t + u = 0, 1, 2, 3 or 4; R9, R10, R11 and R12 represent independently of one another hydrogen atom or (C1-C6)-alkyl; Q represents >N-R13 or compound of the formula: , wherein o = 0, 1 or 2; T represents -N(R15)(R16) or hydroxyl; R13, R15 and R16 represent independently of one another hydrogen atom or (C1-C6); R14 represents hydrogen atom; G represents compounds of the formula or wherein R17, R18, R19, R20 and R21 represent independently of one another hydrogen atom or aryl; J represents compounds of the formula or wherein R22, R23, R24, R25 and R26 represent independently of one another hydrogen or halogen atom; a = 0, 1 or 2; b = 0, 1 or 2; c = 0, 1 or 2; d = 0 or 1; e = 0, 1, 2 or 3; f = 0 or 1; R5 represents hydrogen atom; R6 and R7 represent independently of one another hydrogen atom or (C1-C6)-alkyl; or R6 and R7 can form -(CH2)i-U-(CH2)j- optionally wherein i and j = 1, 2 or 3 independently one another, and U represent a valence bond; R8 represents hydrogen atom or (C1-C6)-alkyl; M represents arylene or -CR27=CR28- wherein R27 and R28 represent independently one another hydrogen atom or (C1-C6)-alkyl. Also, invention relates to a pharmaceutical composition comprising compounds of the formula (I) or its pharmaceutically acceptable salt as an active component in common with a pharmaceutically acceptable carrier or excipient. Also, invention relates to a method for stimulation of growth hormone secretion from mammalian hypophysis that involves administration to mammal the effective dose of compound or its pharmaceutically acceptable salt. Invention provides synthesis of novel compounds and their composition containing thereof that can be used in treatment of diseases associated with the growth hormone deficiency.

EFFECT: improve stimulating method, valuable medicinal properties of compounds and pharmaceutical composition.

13 cl, 29 ex

FIELD: organic chemistry, peptides, biochemistry.

SUBSTANCE: invention relates to compounds of the formula (I): , wherein R means hydrogen atom; R1 means carbocyclic aryl substituted optionally with halogen atom or (carbocyclic aryl or biaryl)-(lower)-alkyl substituted optionally with trifluoromethyl group or halogen atom; alk means (lower)-alkylene; R3 means hydrogen atom or acyl substituted optionally with alkoxy-group, morpholinyl, triazolyl or piperazinyl groups; R4 means hydrogen atom substituted optionally with methoxy-(lower)-alkyl or oxacycloalkyl group; R5 means hydrogen atom or (lower)-alkyl; R6 means (lower)-alkyl; R7 means (lower)-alkyl; or R6 and R7 in common with carbon atom to which they are bound mean 3-10-membered cycloalkylidene; X means -O-,-S(O)n-, -NHCO- wherein n = 0 or 2; -COOR2 means carboxyl or derivative of carboxyl group as a pharmaceutically acceptable ester wherein R2 is chosen from (lower)-alkyl, morpholine carbonylmethyl, dimethylaminocarbonylmethyl or diethylaminocarbonylmethyl, or its pharmaceutically acceptable salt. Proposed compounds are used as dual inhibitors of angiotensin-converting enzyme and neutral endopeptidase and as inhibitor of endothelin-converting enzyme also.

EFFECT: valuable medicinal and biochemical properties of dipeptides.

8 cl, 7 ex

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