N-2-(4-nitrophenyloctyl)etoxycarbonyl-amino acids as n-protected amino acids for solid-phase synthesis of peptides

 

(57) Abstract:

Usage: as N- protected amino acids for solid-phase synthesis of peptides, the Essence of the invention: product: N-2- /4-nitrophenyloctyl/ethoxycarbonylmethylene: 4-NO2-C6H4-S/O/2-CH2-CH2-O-C /O/ - NR1-CHR2-COOH, where R1is hydrogen; R2is hydrogen, methyl, isopropyl, methylpropyl, tert.-butoxymethyl, 1-tert-butoxyethyl/ 2-methylthioethyl, benzyl, carboxamidates, tert-butoxycarbonylmethyl, or together form a propylene radical. Outputs up to 86%. Reagent 1: the corresponding amino acid, Reagent 2: 2-/4 - nitrophenyloctyl/ethylchloride. Reaction conditions: aqueous organic solvent, in the presence of a base at a temperature 0 - 40oC. table 2.

The present invention relates to new derivatives of amino acids having application in the chemistry of peptides, namely, N-2- (4-nitrophenyloctyl)etoxycarbonyl-amino acids of General formula I:

< / BR>
where

R1is a hydrogen atom, and R2takes the following values: hydrogen, methyl, isopropyl, 1-materail, 2-methylpropyl, rubs butoxymethyl, 1-tert-butoxyethyl, 2-methylthioethyl, benzyl, carboxamido the Tyl, 4-tert-butoxybenzoyl, indolyl-3-methyl -, S-(triphenylmethyl)thiomethyl, 1 (triphenylmethyl)imidazolyl-4-methyl, 3-NGmesitylenesulfonyl)propyl, N-cancellability, 2-(N-canterburykeith)ethyl, S -(atsetamidometil)thiomethyl;

or R1and R2together form a propylene radical, is used as the Nprotected amino acids for solid-phase synthesis of peptides.

Solid phase synthesis is widely used for the production of biologically active peptides used in medicine, veterinary medicine, biotechnology, and scientific research.

The essence of solid-phase synthesis of peptides consists in a stepwise extension of the peptide chain through repeated cycles of chemical reactions, starting with the C-terminal amino acids, fixed on an insoluble carrier. The target products of all reactions in the synthesis process remain associated with the media, and the excess reagents and by-products are removed by filtering and washing media.

For the solid-phase synthesis of peptide C-terminal amino acid of the target amino acid sequence, protected a-amino group through a-carboxyl group covalently bind with n-protected aminoacyl-polymer otscheplaut N-protective group and get the aminoacyl-polymer with a free a-amino group. Further, this polymer acelerou the following amino-protected a-amino group, and get N-protected dipeptidyl-polymer. Synthetic cycles, consisting of the stages of the removal of N-protective group and acylation of the free amino group of the peptidyl-polymer, followed by amino-protected a-amino group, repeat until, while not collected the full amino acid sequence of the target peptide.

As in the process of solid-phase synthesis are used in a large molar excess alleluya reagents (2-10-fold compared to free amino groups), all of the reactive groups in the side radicals of amino acids, in particular amino-, carboxy-, hydroxy-, mercapto-, guanidino groups must be blocked by protective groups. Protective groups for this purpose are selected so that, on the one hand, to ensure the full and permanent protection of side radicals of amino acids in the conditions of acylation reactions peptidyl-resin and removal of temporary N-protective group and, on the other hand, to be able to quantitatively and without damage to the structure sintezirovannogo is Alenia connection peptide-polymer. It is obvious that the structure and chemical properties of permanent protective groups of the side radicals of amino acids largely determined by the structure and properties used temporary N-protective group.

Known and widely used in solid-phase synthesis of peptides N-tert-butyl-oxycarbonyl-amenability (BOC-amino) [1] Tert-butyloxycarbonyl (BOC) group is cleaved under the action of acid reagents average power, for example, triperoxonane acid and its solutions in methylene chloride, solutions of hydrogen chloride in organic solvents, epirate boron TRIFLUORIDE and other acids (scheme 1).

< / BR>
Most often in solid-phase synthesis of peptides removal of the BOC-protecting apply processing peptide-polymer 50% solution triperoxonane acid in methylene chloride for 10 to 30 minutes

In combination with temporary N-Boc group for the permanent protection of side radicals of amino acids used protective group, sustainable in terms of removal of Boc-group and split a stronger acid reagents simultaneously with the cleavage of the peptidyl polymer of communication. As such reagents used liquid fluoride is om strategy solid-phase synthesis with N-Boc-semitists using acidolysis for removal and temporary and permanent protective groups that may not provide full stability constant protection. With increasing length of the synthesized peptide permanent protective groups are all more prolonged exposure to the acid that leads to their partial removal. In addition, the processing superstrong acids with the full release of the synthesized peptide causes a number of adverse reactions and leads to partial destruction of the target product. It should also be noted that working with liquid hydrogen fluoride requires special equipment and special security measures.

Application in solid-phase synthesis of peptides N-diciasettenni-amino acids (Dts-amino acids) [2] allows us to overcome a number of disadvantages inherent in the above Boc-amino acids. N-Diseaseclinical (Dts), a protective group is cleaved under the action of tylnej reagents, for example, 2-mercaptoethanol, ethicial-1,2, -2-dipyridine, in a neutral environment with the liberation of the amino group and the formation of therookie carbon (scheme 2).

< / BR>
Conditions of cleavage of the N-Dts-allow you to use it in combination with postanawiaj, close to the cleavage of Boc group, for example, triperoxonane acid and its solutions. In this case, temporary and permanent protective group hatshepsuts by fundamentally different mechanisms. This scheme is called "orthogonal" [3] enables selective removal of temporary N-protective group in the presence of constant (and Vice versa) and, thus, provides full stability constant protection during solid-phase synthesis.

However, the methods of obtaining the N-Dts-amino acid multi-phase and labor-intensive, so they are difficult, expensive and almost not used in practical solid-phase synthesis of peptides.

Known Nderivatives of amino acids used for solid-phase synthesis of peptides, most similar to the proposed compounds according to their chemical properties and method of application are N-9-fluorenylmethoxycarbonyl-amino acids (Fmoc-amino acids) [4] N-9-Fluorenylmethoxycarbonyl (Fmoc) group resistant to acidic reagents and cleaved by the mechanism of b-elimination of organic bases in aprotic solvents, for example, morpholine, diethylamine, piperazine, Alpena and CO2(Scheme 3).

< / BR>
In solid-phase peptide synthesis Fmoc cleavage group mainly uses the processing of the protected peptidyl-polymer mixtures of piperidine with dimethylformamide containing from 20 to 50% piperidine in volume, for 10 to 30 minutes Conditions removal of Fmoc groups also allow you to use it in combination with permanent protective groups of the tert-Putilkovo type atmasamyama soft acidic reagents, which provides the "orthogonality" of the whole scheme of solid-phase synthesis. The flexible off time N-Fmoc-group and permanent protection tert-Putilkovo type provide, generally, high yields and purity of the synthesized peptide.

N-Fmoc-amino acids are widely used in the manual solid-phase peptide synthesis, and automatic and semi-automatic synthesizers reactor shaker and column types. However, due to the relatively high value of N-Fmoc-amino acids, they are used only in the synthesis of small scale (usually no more than 2 to 3 mmol of C-terminal amino acid). In addition, due to the high lability of N-Fmoc-amino acids in the presence of even weak bases, they are rarely applied for the synthesis of peptides with a length of the N-protected derivatives of amino acids, biodegradable organic bases in non-aqueous environments and is suitable for solid-phase synthesis of peptides.

This objective is achieved in that the features that are offered new compounds, namely N-2-(4-nitrophenyloctyl)etoxycarbonyl-amino acid N-Nsc-aminokisloty) of General formula I:

< / BR>
where

R1is a hydrogen atom, and R2takes the following values: hydrogen, methyl, isopropyl, 1-methylpropyl, 2-methylpropyl, tert-butoxymethyl, 1-tert - butoxyethyl-methylthioethyl, benzyl, carboxymethyl, 2-carboxamidates, tert-butoxycarbonylmethyl, 2 (tert-butoxycarbonyl)ethyl, 4-(tert-butoxycarbonyl)butyl, 4-tert-butoxybenzoyl, indolyl-3 - methyl -, S-(triphenylmethyl)thiomethyl, 1-(triphenylmethyl)imidazolyl-4 - methyl, 3-(NG-mesitylenesulfonyl)propyl, N-cancellability, 2 - (N-canterburykeith)ethyl, S-(atsetamidometil)thiomethyl;

or R1and R2together form a propylene radical,

as N-protected amino acids for solid-phase synthesis of peptides.

These compounds can be obtained by treating the amino acids of General formula II, where R1and R2oC (scheme 4).

< / BR>
Chloroformiate III is introduced into the reaction in an amount of from 0.5 to 1.5 g-EQ. preferably 0.7 to 0.9 g-EQ. with respect to the amino acid. As the organic component of the solvent take aprotic organic solvent, dissolving allerease agent and miscible with water, for example, acetonitrile, dimethylformamide, tetrahydrofuran, dioxane. As the base used inorganic or organic base, such as potassium carbonate or sodium, magnesium oxide or calcium, triethylamine, N-methylmorpholine.

According to another method, the amino acid of formula II in a known manner into N, O-bis-trimethylsilyl derivative of the formula IV, which are then without releasing pure process chloroformiate III in the environment anhydrous organic solvent, for example methylene chloride. After aqueous hydrolysis of the intermediate trimethylsilyl esters receive free N-Nsc-amino acids of formula I (scheme 5).

< / BR>
The compounds of formula I, where R1a hydrogen atom, and R2- N-cancellability or 2(N-canterburykeith)ethyl, produced by interaction of the compounds of formula I, where R1carboxamidates or 2-(carboxamido)ethyl, Sohotel can be used dimethylformamide, the acid is preferably an organic acid, for example, triperoxonane acid, n toluensulfonate or methansulfonate.

From the structural formula shows that the compounds of formula I have asymmetric a-carbon atom (except in connection with R1=R2=H). Because the reactions used to produce it, is not involved, the a-carbon atom, the configuration of this atom present in the initial amino acid of the formula II, is stored in N-Nsc-amino acids of formula I. Thus, it is obvious that, depending on the configuration of the original amino acid of formula II described methods can be obtained N-Nsc-amino acids D - or L-configuration, as well as racemic compounds.

Values of the substituents R1and R2in the inventive compounds of the formula I correspond to the structures of side radicals of amino acids of natural origin containing or not containing protective groups known structure, mainly tributylin type or similar conditions removal (table 1).

From table 1 it is evident that the compounds of formula I constitute a complete set of protected derivatives of amino acids required for the synthesis of peptides of any minocycline or manorastroman in water and soluble in polar organic solvents, stable during prolonged storage at temperatures from -10 to 25oC.

N-Nsc-amino acids I applied for hardening the synthesis of peptides. To this end, the first N-Nsc-amino acid corresponding to the C-terminal amino acid of the amino acid sequence of the target peptide, through the a - carboxyl group covalently linked to an insoluble polymer carrier via the formation of amide or ether linkages. When this occurs, the formation of N-Nsc-aminoacyl-polymer. As the polymer carrier use mesh or macroporous polystyrene crosslinked with divinylbenzene, poly-N,N - dimethylacrylamide in the form of granules or in the form of a composite with diatomaceous earth, cross-linked dextrans, cellulose, paper, and other polymeric carriers, known for solid synthesis of peptides. Polymeric carrier must contain an anchor group required for attaching the first amino acid. The preferred anchor groups are groups that provide cleavage of the synthesized peptide from the polymer carrier with the release of C-terminal carboxyl or carboxamide group when treatment with acidic reagents, for example, triperoxonane acid, solution hydrogen chloride in lilina, 4-chloro - or 4-bromatological, a-hydroxydiphenylmethyl and other well-known anchor group; for the formation of amide linkages known di - and dialkoxybenzene groups with different location alkoxy-substituents, 4-aminomethyl-3,5-dimethoxyphenethylamine, as well as other well-known group used for this purpose.

Joining C-terminal Nsc-amino acids to the anchor groups of the polymer carrier is carried out by methods known in the chemistry of peptides.

To obtain the aminoacyl-polymer with a free a-amino group with the NNsc-aminoacyl-polymer otscheplaut Nsc group. To this end, the polymer is treated with a basic reagent. Preferred basic reagents for this purpose are nitrogenous bases, such as ammonia, morpholine, piperidine, piperazine, diethylamine, 1,8-diazabicyclo[5,4,0]under-7-ene, 1,1,3,3-tetramethylguanidine and their solutions in aprotic organic solvents. The most preferred basic reagent is a solution of from 20 to 50% by volume of piperidine in dimethylformamide. The cleavage products Nsc-groups in this case are N-[2-(4-nitrophenyloctyl) ethyl]piperidine and CO2with the release of the a-and the-Nsc-amino acids and receive N-Nsc-dipeptidyl-polymer. To this end, apply the methods and techniques known in the chemistry of peptides. As alleluya reagents can be used 4-nitrophenolate, pentachlorophenolate, pentafluorophenyl, 1-oxybenzenesulfonate esters of N-Nsc-amino acids, as well as other types of activated esters used in solid-phase synthesis of peptides; symmetric anhydrides of N-Nsc-amino acids. The acylation can be performed N-Nsc-amino acids in the presence of a known condensing reagents, for example, dicyclohexylcarbodiimide, diisopropylcarbodiimide, hexaflurophosphate, benzotriazolyl-1-oxy-Tris(dimethylamino)phosphonium.

Synthetic cycles, consisting of the stages of the removal of N-Nsc-group and acylation of the free amino group of the peptidyl-polymer subsequent N-Nsc-amino acid until then, until it is collected amino acid sequence of the target peptide.

At the end of the Assembly with the N-Nsc-peptidyl polymer is removed N-Nsc-group, then the target peptide otscheplaut from the anchor group of the polymer with simultaneous removal of permanent protective groups of the side of the amino acid radicals, for example triperoxonane acid solutions methanesulfonate or p-toluenesulfonic acid in organic solvents, with or without adding known reagents for binding formed Karbalevich ions, for example, anisole, thioanisole, dimethyl sulfide, ethicial-1,2.

The released target peptide is isolated and purified by known methods.

N-Nsc-group slower than N-Fmoc-group, split by reason, however, for real-time, which is usually given then the solid-phase synthesis Protocol for the stage of removal of N-protective group (15 - 20 min), is, however, the quantitative removal of Nsc-group with the peptidyl-resin. However, the greater stability of N-Nsc-amino acids by the action of bases compared with Fmoc-amino acids provides them more stable in neutral and weakly basic media used for carrying out the stages of acylation.

As stated above, the Nsc group can be quantitatively derived key reagents in the presence of the protective groups of the tert-Putilkovo type, which is resistant to the action of organic bases. On the other hand, Nsc-group fully resistant to the action is use for solid-phase synthesis of peptides N-Nsc-amino acids of formula I containing radicals in the side permanent protective group mainly tert-Putilkovo type or similar conditions removal, helps to ensure the conditions of orthogonality, which are the basis of effective schemes of solid-phase synthesis of peptides.

The essence of the proposed invention is illustrated by examples. In the description of examples, the following abbreviations and symbols:

DMF dimethylformamide,

HPLC high performance liquid chromatography.

Optically active amino acids listed in the descriptions of the examples, by default, have the L-configuration.

Example 1. Obtaining N-Nsc-L-asparagine (I 10).

Dissolved 3,96 g (30 mmol) of L-asparagine and 7.7 g (55 mmol) of potassium carbonate in 100 ml of a mixture of water-dioxane (3:1) and cooled in an ice bath for 15 min was added with stirring, a solution of 7.5 g (25 mmol (2-(4-nitrophenyloctyl)ethylchloride III in 70 ml of dioxane. The mixture was stirred for another 20 min without cooling, evaporated under reduced pressure to 100 ml and transferred into a separating funnel. To the mixture was added 100 Il of water and was extracted with ethyl acetate (2 x 50 ml). The aqueous layer was separated, acidified with 40% of CE is th until neutral, was air-dried until constant weight. Received connection 1 10 in the form of white crystalline powder, yield 71% characteristics of the obtained product are shown in table 2 (Example 5).

Example 2. Obtaining N-Nsc-L-leucine (1 5).

to 4.92 g (37.5 mmol) of L-leucine and 90 ml of dry methylene chloride was placed in a round bottom flask with a capacity of 250 ml, equipped with a reflux condenser and addition funnel. To the suspension with vigorous stirring was added to 9.5 ml (75 mmol) of trimethylchlorosilane, the mixture was heated to boiling and boil 1 hour the mixture is Then cooled to Delaney bath and added with stirring to 9.1 ml (65 mmol) of triethylamine and 9.0 g (30 mmol) of chloroformiate III. The mixture was stirred for 20 minutes under cooling, then 1.5 h at room temperature. The solvent was distilled on a rotary evaporator and the residue is distributed between 200 ml ethyl acetate and 250 ml of 2.5% sodium bicarbonate solution. The aqueous layer was separated, washed with 50 ml of ether, acidified using 1 N. hydrochloric acid to pH 2 and extracted with ethyl acetate (200 ml). An ethyl acetate extracts were dried with sodium sulfate, the solvent was removed, the residue was recrystallize from a mixture of hexane-ethyl acetate. Received connection 1 5 in the form of white crystalline powder, yield 80% Features floor is c-L-aspartic acid (1 12).

to 7.09 g (37.5 mmol) of b-tert-butyl ester of L-aspartic acid and 90 ml of dry methylene chloride was placed in a round bottom flask with a capacity of 250 ml, equipped with a reflux condenser and addition funnel. To the mixture with vigorous stirring was added a 12.7 ml (73 mmol) of diisopropylethylamine, then to 9.5 ml( 75 mmol) of trimethylchlorosilane and boiled reactions mixture of 1.5 hours the Mixture was cooled in an ice bath, was added at once 7.9 g (30 mmol) of chloroformiate III and stirred 1.5 h at room temperature. The solvent was distilled on a rotary evaporator and the residue is distributed between 200 ml ethyl acetate and 250 ml of 2.5% sodium bicarbonate solution. The aqueous layer was washed with 50 ml of ether, acidified using 1 N. hydrochloric acid to pH 2 and extracted with ethyl acetate (200 ml). An ethyl acetate extracts were dried with sodium sulfate, the solvent was removed, the residue was recrystallized from a mixture of hexane-ethyl acetate. Received connection 1 12 in the form of white crystalline powder, yield 86% characteristics of the obtained product are shown in table. 2 (Example 5).

Example 4. Obtaining N-Nsc-N-xanthyl-L-asparagine (1-20).

Dissolved to 3.89 g (10 mmol) of N-Nsc-L-asparagine (1 10) and 2.6 g (13 mmol) of xanthydrol in 20 ml of DMF. To the solution was added 0.4 ml metasolv 100 ml of ice water, the precipitation was filtered, washed with water, ethyl acetate, ethanol, ether. The precipitate was dissolved by heating in 10 ml of DMF, was filtered, was besieged with ether, washed with ether, and dried in a vacuum exicator over calcium chloride. Received connection 1 20 in the form of a crystalline powder, yield 74% characteristics of the obtained product are shown in table. 2 (example 5).

Example 5. Physico-chemical properties of N-Nsc-amino acids I.

Using the methods described in examples 1 to 4, the compounds of formula I, are given in table. 2. The number in the column "Method" represents the number of the example, which sets forth the method of obtaining. Value of specific optical rotation []2D5measured on a polarimeter DIP-320 JASCO (Japan) in a cell with a length of 10 see the melting Temperature measured in capillaries and is not corrected. The values of the chromatographic mobility of Rfrefer to plates for thin-walled chromatography Alufolien Kieselgel 60 F254(Merck, Germany) in the system chloroform-methanol-Oksana acid, 95:5:3 (a) and benzene-acetone-Oksana acid, 100:50:3 (B). Detection of the spots on the plates was performed in UV-light and ninhydrin reagent after warming up. The masses of the molecular ions (M+H+is soedinenii formula I corresponds to computing within the error of the method definition (CHN-analyzer Perkin-Elmer, the data in the table not given). All optically active compounds I, are shown in table 2, we have L-configuration.

Example 6. Synthesis of dodecapeptide Ala-Ser-Ser-Thr-Ile-Ile-Lys-Phe-Gly-Ile-Asp-Lys.

a) the Introduction of anchor groups in the polymeric carrier.

To 250 mg aminomethylpropanol copolymer of styrene with 1% divinylbenzene (1.0 mEq NH2/g) was added 0.75 mmol 2,4,5-trichlorphenol ether 4-hydroxymethylphosphonate acid, 0.75 mmol of 1-hydroxybenzotriazole and 3 ml of DMF. The reaction mixture was stirred 24 h at room temperature. The polymer was filtered, washed DMF, ethanol, ether, hexane, dried 24 h in a vacuum desiccator over pjatiokisi phosphorus.

b) Attaching Nsc-Lys(Boc)-OH to anchor the group.

To the obtained polymer was added to 0.75 mold Nsc-Lys(Boc)-OH (1 14), 0.75 mmol of dicyclohexylcarbodiimide, 0.1 mmol of 4-dimethylaminopyridine and 4 ml of a mixture of dichloromethane-N-organic (3:1). The reaction mixture was stirred 24 h at room temperature. The polymer was filtered, washed thoroughly on the filter with chloroform, a mixture of chloroform-methanol (1:1), ethanol, ether and hexane. Received 400 ml of Nsc-Lys(Bos)-polymer.

C) assembling the amino acid sequence of dodecapeptide.

200 m the new filter. The polymer in the syringe thoroughly washed DMF and then carried out the synthetic cycles according to the following Protocol:

1. Washing 33% piperidine/ DMF, 4 ml; 0.5 minutes

2. Release: 33% piperidine/DMF, 4 ml; 15 minutes

3. Washing: DMF, 6 x (4 ml; 1 min).

4. Acylation: N-Nsc-amino acid I, 0.5 mmol; hexaphosphate benzotriazolyl-1-oxy-Tris(dimethylamino)phosphonium (THIEF), 0.5 mmol; 1-hydroxybenzotriazole, 0.5 mmol; N-methylmorpholine 0.75 mmol; DMF, 2 ml; 60 min (Nsc-Ile-OH 90 min).

5. Washing: DMF, 5x(4ml; 1 min).

NNsc-Amino acids were introduced into the synthetic cycles in the following sequence: Nsc-Asp(OtBu)-OH, Nsc-Ile-OH, Nsc-Gly-OH, Nsc-Phe-OH, Nsc-Lys(Boc(-OH, Nsc-Ile-OH, Nsc-Ile-OH,Nsc-Thr(tBu)-OH, Nsc-Ser(tBu)-OH, Nsc-Ser(tBu)-OH, Nsc-Ala-OH,

After Assembly, the target amino acid sequence peptidyl-polymer was treated with 33% piperidine in DMF (4 ml) for 20 min, then washed DMF, methylene chloride, ethanol, ether, hexane.

g) releasing and clearing dodecapeptide.

Peptidyl-polymer was treated with 5 ml of 50% triperoxonane acid in dichloroethane with stirring for 60 min at room temperature. The polymer was filtered, washed on the filter with 5 ml of 50% triperoxonane Kalevala, was washed with ether and dried in vacuum. Obtained 170 mg of crude dodecapeptide 70% purity according to analytical reversed-phase HPLC.

Crude dodecapeptide dissolved in 3 ml of 1 M aqueous solution of acetic acid and was chromatographically on column 1,h cm TSK HW-40F(Merck, Germany ), equilibrated 1 M aqueous solution of acetic acid. The elution was performed with the same buffer, and fractions containing pure peptide were combined and liofilizovane. Received 104 mg (41%) target dodecapeptide with a purity of more than 95% according to analytical reversed-phase HPLC. The analysis of amino acids after hydrolysis of 6 N. HCl, 110oC 24 and 48 h relative to Ala): Asp 1.02 (1); Ser 1,84 (2); Thr 0,93 (1); Glu 0,94(1); Gly 1.03(1); Ala 1.00 (1); Ile 2.78(3); Lys 2.04 (2).

These examples show that the inventive compounds N-2-4(nitrophenyloctyl)etoxycarbonyl-amino acids of General formula 1 can be used as the N-protected derivatives of amino acids in solid-phase synthesis of peptides.

N-2(4-Nitrophenyloctyl) etoxycarbonyl-amino acids of formula I

< / BR>
where R1hydrogen;

R2hydrogen, methyl, isopropyl, 1-methylpropyl, 2-methylpropyl, tert-butoxymethyl, 1-tert-butoxyethyl, 2-methylthioethyl, benzyl, carboxin is about)butyl, 4-tert-butoxybenzoyl, indolyl-3-methyl -, S-(triphenylmethyl)thiomethyl, 1-(triphenylmethyl)imidazolyl-4 - methyl, 3-(NG-mesitylenesulfonyl) propyl, N-cancellability, 2-(N-canterburykeith)ethyl or S-(atsetamidometil)thiomethyl

or R1and R2together form a propylene radical,

as Nprotected amino acids for solid-phase synthesis of peptides.

 

Same patents:

The invention relates to the field of biotechnology and relates to a method of obtaining ovomucoid (Ω) of the whole protein eggs (CBA)

The invention relates to the field of biotechnology and relates to a method of obtaining ovomucoid (Ω) of the whole protein eggs (CBA)

The invention relates to organic chemistry, in particular to preparative peptide synthesis of GnRH analogues in solution

The invention relates to the processing of vegetable raw materials, namely the method of obtaining protein hydrolysates, for example, from soybean meal and algal waste agar production, which can be used for food or as supplements in animal feed
The invention relates to biologically active substances and can be used to obtain products with a high physiological activity as a form of application of amino acids in the preparations of clinical nutrition

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to a method for preparing acid-additive salts of compounds of the formula (I):

wherein R1, R2 and R3 represent alkyl comprising from 1 to 12 carbon atoms. Substances of the formula (I) are inhibitors of cellular Na+/H+ antiporter and therefore can be used in treatment of arrhythmia arising from oxygen insufficiency. Method involves the following stages: alkyl-derivatives of 4-chloro-5-methanesulfonylbenzoic acid are converted to ester that is subjected for reaction with alkyl sulfinate, and prepared substance is treated with guanidine followed by preparing acid-additive salts of compound of the formula (I). Method provides the best yield of the end product and simplified the technology.

EFFECT: improved preparing method.

10 cl, 3 ex

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to fluorinated cycloalkyl-substituted benzoylguanidines of the formula (I) wherein X means oxygen or sulfur atom or -NR6 wherein R6 means hydrogen atom, alkyl comprising 1, 2, 3 or 4 carbon atoms; m = 0, 1, 2 or 3; n = 0, 1, 2 or 3; p = 0, 1, 2 or 3; q = 1, 2 or 3; r = 0, 1, 2 or 3 and wherein sum m + n + p + q + r is 2 at least; R1 means hydrogen atom or alkyl comprising 1, 2, 3 or 4 carbon atoms; R2 means hydrogen atom or alkyl comprising 1, 2, 3 or 4 carbon atoms; R3 means -SOuR10 wherein u means 0, 1 or 2; R10 means alkyl comprising 1, 2, 3 or 4 carbon atoms; R4 means hydrogen atom or alkyl comprising 1, 2, 3 or 4 carbon atoms; R5 means hydrogen, and their pharmaceutically tolerant salts. Also, invention relates to compounds of the formula (I) and/or their pharmaceutically tolerant salt used as a medicinal agent possessing inhibitory effect on activity of sodium-proton antiporters and designated for treatment of diseases mediated by this activity. Invention provides fluorinated cycloalkyl-substituted benzoylguanidines possessing inhibitory activity with respect to sodium-proton antiporters.

EFFECT: valuable medicinal properties of compound and medicinal agents, improved method of synthesis.

9 cl, 2 ex

FIELD: organic chemistry, chemical technology, biochemistry, pharmacy.

SUBSTANCE: invention relates to compounds of the formula (I): wherein both X1 and X2 represent methylene; R3 represents -CR5=CHR6, and R5 and R6 in common with atoms to which R5 and R6 are bound form (C6-C12)-aryl wherein R3 is substituted optionally with 1-5 radicals of the formula: -X4OR9 wherein X4 represents a bond; R9 represents halogen-substituted (C1-C3)-alkyl, and R4 represents -C(O)X5R11 wherein X5 represents a bond, and R11 represents hetero-(C6-C6)-cycloalkyl-(C0-3)-alkyl; X3 represents group of formulae (a) , (b) or (c) wherein n = 0, 1 or 2; R20 represents hydrogen atom (H); R21 is chosen from group consisting of H, -C(O)R26, -S(O)2R26 wherein R23 is chosen from H and (C6-C12)-aryl-(C0-C6)-alkyl; R25 is chosen from H, (C6-C12)-aryl-(C0-C6)-alkyl or -X4S(O)2R26 wherein X4 has above given values; R26 is chosen from group consisting of H, (C6-C12)-aryl-(C0-C6)-alkyl; wherein X3 comprises optionally, except for, one substitute that being in alicyclic or in aromatic ring system represents a radical chosen independently from group consisting of -X6OR17 wherein R17 represents H, (C1-C6)-alkyl, and X represents a bond or (C1-C6)-alkylene; and its N-oxide derivatives, protected derivatives, individual isomers and mixtures of these isomers; and pharmaceutically acceptable salts and solvated of such compounds, its N- oxide derivatives, protected derivatives, individual isomers and mixtures of these isomers. Also, invention describes a pharmaceutical composition possessing inhibitory activity with respect to cathepsin S-proteases based on compounds of the formula (I), and compound of the formula (Ix) given in the invention description. Invention provides preparing novel compounds possessing useful biological properties.

EFFECT: improved preparing method, valuable medicinal and biological properties of compounds and pharmaceutical composition.

16 cl, 3 tbl, 17 ex

FIELD: chemistry.

SUBSTANCE: in formula is or , is , R1 is hydrogen, halogen, nitro or amino, R2 is hydrogen or lower alkyl, R3 is hydrogen, -X- is where -Y- is a bond, -O-, -NH- or -CH2-, and R4, R5 and R6 each denotes hydrogen, R7 is (C1-C8)-alkyl, (C3-C8)-cycloalkyl, -Z-R9 or , where -Z- is -O- or -S-, and each R9 independently denotes (C1-C8)-alkyl, (C3-C8)-cycloalkyl, and R8 is -D-E-R10, in which -D- is -CONHSO2-, E is (C1-C6)-alkylene and R10 denotes -O-R11 in which R11 is hydrogen. The invention also relates to use of said compounds to prepare a medicinal agent, a pharmaceutical composition and a method of preventive and/or therapeutic treatment of overactive bladder and/or acraturesis.

EFFECT: derivatives are highly effective.

7 cl, 56 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to 2-N-halogen-4-methylsulphonyl-butyric acid derivative or to its pharmaceutically acceptable salt , wherein X=F, Cl or Br. The invention also refers to a method for preparing the above compound and to a pharmaceutical composition for treating an inflammation on the basis of the above compound.

EFFECT: there are prepared a new compound and a pharmaceutical composition on its basis which can find application in medicine for treating inflammation or disorders referred to inflammation, bacterial infection, pain or skin conditions.

15 cl, 2 dwg, 1 tbl, 11 ex

FIELD: pharmaceutics.

SUBSTANCE: invention relates to an agent exhibiting anti-influenza activity, which is a compound of general formula (I) in form of a mixture of R- and S-diastereomers or separate isomers or pharmaceutically acceptable salts. Invention also relates to use of compounds of general formula (I).

EFFECT: invention can be used as an agent exhibiting anti-influenza activity.

3 cl, 2 tbl, 8 ex

FIELD: biotechnology, biochemistry.

SUBSTANCE: invention relates to extracts prepared from vegetable somatic embryos for the cell-free translation system and/or the coupled transcription-translation system. Method involves preparing embryonic callus from the primary material and the embryonic suspension culture. After induction of the secondary somatic embryogenesis extract is prepared from somatic embryos. Based on the extract the diagnostic system is developed for detection of biologically active compounds. Invention provides overcoming the species limitations and strain specificity and to attain the high effectiveness of the cell-free translation system and the coupled transcription-translation system also.

EFFECT: improved preparing method, valuable biological and biochemical properties of system.

49 cl, 5 dwg, 2 tbl, 9 ex

FIELD: bioengineering; genetic engineering; medicine; methods of production casamino acids.

SUBSTANCE: the invention is pertaining to the field of bioengineering, genetic engineering, medicine, in particular, to the methods of production of components for nutrient mediums from hydrolysates of animal protein. The invention offers the method of production of casamino acids by the method of the gel permeation chromatography of the hydrolyzed crude acid casein with the contents of the general nitrogen - 0.7-0.95 g in 100 ml of the solution and concentration - 6-10 % on Sefadex G-15, eluating by a distilled water of fractions of an eluate, selection of the active fractions of an eluate by a spectophotometery of portions of the eluate (D254), evaporation of the active fractions under vacuum at the temperature of no more than 55°C. The method allows to simplify the process of production of casamino acids, to reduce its cost and also to obtain casamino acids possessing the high growth- stimulating activity.

EFFECT: the invention ensures simplification of the process of production of casamino acids, reduction of its cost and also production of casamino acids possessing the high growth- stimulating activity.

2 cl, 2 dwg, 1 tbl, 1 ex

FIELD: peptides, pharmacy.

SUBSTANCE: invention relates to a new method for preparing a pharmaceutical composition for the parenteral administration in mammals that comprises salt of difficulty soluble peptides. Method involves treatment of the parent readily soluble acid-additive salt of the basic peptide of LHRH antagonist in the presence of suitable diluting agent with the mixed ion-exchange resin or mixture of acid and basic ion-exchange resins to form free basic peptide. Then ion-exchange resin is removed and free basic peptide is treated with inorganic or organic acid to form the final product, required acid-additive peptide salt followed by addition of suitable pharmaceutical vehicles and/or filling agents and removal of a diluting agent.

EFFECT: improved preparing method.

5 cl, 1 dwg, 1 ex

FIELD: feed mill industry.

SUBSTANCE: method involves treating sunflower oilcake with catholyte; removing treated solution; extracting protein; filtering; drying sediment; grinding; simultaneously with treatment of sunflower oilcake with catholyte, providing treatment of soya with anolyte, with catholyte and anolyte circulating at equal velocities. Apparatus has two chambers connected with each other through semi-permeable partition. Each of said chambers is equipped with electrodes and dc source. Apparatus is further equipped with collecting chamber and pumps with pipelines.

EFFECT: increased efficiency of method and apparatus, reduced production time and decreased costs of additives.

1 dwg, 1 tbl

Up!