Cyclic peptides or their additive salt of the acid, the composition exhibiting antifungal and antipneumococcal activity, a method of treating fungal infections

 

(57) Abstract:

The invention relates to certain Aza cyclopentapeptide compounds having a nitrogen atom attached to cyclohexadienone ring on the 5th carbon atom of the component 4-hydroxy-ornithine (C-5-orn"), formula I, where R1Is H or OH; R2- H, CH3or OH; R3- H, CH3CH2CN, CH2CH2NH2or CH2CONH2; RI- C9-C21-alkyl, C9-C21alkenyl, C1-C10-alkoxyphenyl or C1-C10alkoxyaryl; RII- H, C1-C4-alkyl, C3-C4alkenyl, (CH2)2-4OH, CO(CH2)1-4NH2, (CH2)2-4NRIVRV; RIIand RIIItaken together, -(CH2)4-, -(CH2)5-, -(CH2)2O(CH2)2-, -(CH2)2NH(CH2)2-; RIV- H or alkyl; RVIs H or alkyl and salt additive. The compounds I are active against many fungi, in particular against Candida species. Compounds of the present invention are also useful for inhibiting or facilitating the flow of infectious diseases caused Pneunocystis carini in patients with reduced immunity. 4 c. and 7 C.p. f-crystals, 10 PL.

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Aza cyclopentapeptide connection, in the present invention, compound I (Seq ID Nos. 1-15), differ in that they have a nitrogen atom attached to cyclohexadienone ring on the 5th carbon atom of the component 4-hydroxy-ornithine (hereinafter referred to as"C-5-orn") and can be represented by the formula

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where

R1Is H or OH;

R2- H, CH3or OH;

R3- H, CH3CH2CN, CH2CH2NH2or CH2CONH2;

RI- C9-C21alkyl, C9-C21alkenyl, C1-C10alkoxyphenyl or C1-C10alkoxyaryl;

RII- H, C1-C4alkyl, C3-C4alkenyl, (CH2)2OH, (CH2)2-4NRIVRV, CO(CH2)1-4NH2;

RIII- H, C1-C4alkyl, C3-C4alkenyl, (CH2)2-4OH, (CH2)2-4NRIVRVor

RIIand

RIIItaken together, - (CH2)4-, -(CH2)5-, -(CH2)2O(CH2)2-, -(CH2)2NH(CH2)2-,

RIV- H or C1-C4alkyl,

RV- H or C1-C4alkyl;

and their additive salt of the acid.

Used here, the expression "alkyl"is soedineniya in the present invention is mainly obtained as mixtures of stereoisomeric forms, which is usually dominated by one form. Selection conditions in order to obtain the target isomer as a dominant is not beyond the normal skills of experienced synthetics. The connection in the preferred stereoisomeric form, shown here as "normal" form, shown in the working examples with dashed lines ties beyond the plane in position C-5-orn". The designation "EPI" is applied to those compounds in which the group in position C-5-orn" is above the plane.

Pharmaceutically applicable salts, obtained as salts of related acids, are formed with such acids as chloromethane, Hydrobromic, phosphoric, sulfuric, maleic, citric, acetic, tartaric, succinea, oxalic acid, malonic acid, glutamic and the like, and include other acids, giving pharmaceutically applicable salts listed in Journal of Pharmaceutical Science, 66, 2, (1977).

Typical kernel for Aza derivatives of the present invention (compound I) and the sequence identifier (Seguence ID) for these compounds are presented in the following table. 1. Because peptide cores are the same, regardless of the substituents RI, RIIor RIIIand because the room identificat the awn.

One of the compounds, particularly stand out from the rest, because it has the ability to fight fungal infections, designated as compound I-6, where RII- H, RIII- CH2CH2NH2and RI- 9,11-dimethylthiazol (TMTD), and which can be marked as compound I-6-1 (Seq. ID NO. 6).

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The above designation I-6-1 refers to the first connection, in which the location of the kernel - I-6. Since all the compounds in the present invention, the substituent in the "C-5-orn" - nitrogen, the substituents on the indicated nitrogen atom may vary, and all connections that have the same R1, R2and R3will refers to Seq ID No. 6.

Compounds soluble in the lower alcohols and polar aprotic solvents such as dimethylformamide (DMF), dimethylsulfoxide (DMSO) and pyridine. They are insoluble in the solvents of the type of diethyl ether and acetonitrile.

Compounds of the present invention is used as antibiotics, especially as fungicides, or as a means of combating the simplest organisms. As fungicides, they are used to combat both filamentous fungi and yeasts. These compounds are particularly adapted for use in the treatment of fungi is occus, such as C. neoformans and, apparently, Aspergillus: A. fumidatus, A. flavus, A. niger. They are also used for the treatment or prevention of Pneumocystis carinii pneumonia, to which patients with low immune system are especially susceptible, as will be discussed in the future.

Compounds of the present invention can be obtained from cyclopeptides formula

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through a series of transformations, in which the oxygen atom at position C-5-orn" (which can also be named as hemiaminal position) eventually replaced by a nitrogen atom. The starting materials may be natural or modified natural products, as will be subsequently described. If R1-hydrogen, but not hydroxyl, Aza compounds may be obtained from another series of reactions. The method is applicable for producing compounds in which R1can be hydrogen, and hydroxyl described first.

The sequence identifiers of the source materials shown in the table. 2.

Compound a-4 and a-7 described in the literature (J. Antibiontcs 45, 1855-60 Dec. 1992) as pneumocandin Boand pneumocandin Aoif RI=DMTT.

When in the compound A-1 R1and R2presents one of the possible substituents is but the first method. If R3- CH2CN or-CH2CH2NH2the group-CH2CONH2first, it may be converted into a-CH2CN or-CH2CH2NH2as it is consistently described, and all modified compounds (Seq ID Nos. 17-18, 20-21, 23-24, 28-29) can be used in the first method [see I. 2], or otherwise, the compound in which R3-CH2CONH2can be used to obtain compounds with nitrogen in hemiaminal position, and then CH2CONH2group of the obtained product can be converted into a-CH2CN or-CH2CH2NH2.

If R1, R2and R3in the source material are the same as in the product, it can be used the following sequence of transformations:

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*Position C-5-orn" or hemiaminal position.

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At the stage of A starting material, the compound A (Seq ID Nos. 16-30), alkylthiol or arylthiol and acid react in an aprotic solvent in the absence of moisture for a time sufficient for reaction with the formation of compound B (Seq ID Nos. 31-45), shown in the table. 3. It was found that acylaminoacyl well suited for carrying out this stage. Compound B obtained from soedinenii. Examples of strong organic acids are sulforaphane acid, p-toluensulfonate and methansulfonate. Mineral acids include chloroethanol and Hydrobromic acid. It is preferable sulforaphane acid.

Suitable solvents are DMF, DMSO, 1-methyl-2-pyrrolidone and hexamethylene phosphoric acid (GMAT). Preferred DMF and DMSO.

The reaction in General proceeds at room temperature for 1-10 days.

If the reaction cyclopentapeptide connection, thiol and acid are mixed in a suitable solvent before the end of the transformation. The reaction mixture was diluted with water and hold brasindiana flash chromatography, using as eluent 10-40% solution of acetonitrile in water containing 0.1% triperoxonane acid. Triperoxonane acid hereinafter referred to as TFUCK. The fraction containing the target product can be concentrated and lyophilized, and the lyophilized material can be purified preparative high performance liquid chromatography (HPLC).

Suitable columns for HPLC are sold under trademarks or trade names of "BOND" (DuP is Tadei B connection C, sulfon (Seq ID Nos. 31-45) is obtained by oxidation of compound B. Suitable oxidizing agents are "OXONE" (KHSO5: KHSO4K2SO4= 2: 1:1, Aldrich Chemicals), m-chloro-peroxybenzoate acid and peroxidasa acid. The sequence identifier (Seq ID) for the connection C is the same as for B, because the atom attached to hemiaminals carbon remains a sulfur atom. Thus, the sequence identifiers (Seq ID) for the sulfones are as follows (see tab. 4).

Oxidation of tiefer (compound B) to the sulfone (compound C) is approximately two moles of oxidant. When using one mole of the oxidizing agent the reaction product is a sulfoxide, which may be further converted into a sulfon. In the formation of Aza compounds as intermediates can be used sulfoxidov, but preferable to the use of sulfones. Use a slight excess of oxidant over dumalinao number.

The reaction is carried out in the aquatic environment, mainly in a mixture of acetonitrile with water. Preferably about the same grade, although you can use a mixture with ratio from 1:9 to 9:1.

When conducting reactivate at room temperature for a time sufficient to complete the reaction of formation of compounds C, typically from 30 min to 1 h

Upon completion of the reaction, the compound isolated from reaction mixture by dilution with water and chromatographytandem. For the stage of purification is suitable brasindiana column flash chromatography (C18). The preferred eluent is 30-45% acetonitrile in water (containing 0.1% TFUC) increments of 5%. The desired fractions lyophilizer for targeted intermediate sulfone, compound C (Seq ID Nos. 31-45). The intermediate product is unstable, so the selection should be done as soon as possible.

Compound C can be converted into a compound having a nitrogen atom directly attached to the C-5-orn". As shown in the diagram of the synthesis, the reaction of compound C with an alkali metal azide leads to the entry of the azide group at that position, whereas the reaction with ammonia or an amine gives the amino group in position C-5-orn" (compound I). Compound D is an important intermediate product for most of the substances according to the present invention. Although the connection D is a nitrogen atom at position C-5-orn", but because it is not a product, it has a separate nomemade in table. 5.

Azide can be obtained by adding an alkali metal azide to a solution of sulfone (compound C; Seq ID Nos. 31-45) in an aprotic solvent with stirring at room temperature for time sufficient to complete the formation of the azide determined by HPLC. After that the reaction mixture can be diluted aqueous acid solution, such as TFUK, and then target azide (compound D) can be isolated from the reaction mixture chromatographically. This is suitable brasindiana column flash chromatography (C18) using 10-25% acetonitrile in water (containing 0.1% TFUC) increments of 5%.

Azide (compound D) can be restored to compounds having a free amino group, which is represented among the products of the present invention (Seq ID Nos. 1-15).

Recovery may be performed by mixing azide (compound I) with palladium deposited on charcoal (Pd/C), solvent type, glacial acetic acid saturated with hydrogen under a pressure equal to the pressure in the supply tank, in 10-20 hours To separate the product first, the catalyst is filtered off and the filtrate lyophilizer to obtain aminated the ü converted into a substituted amine, as described next.

Compound I, in which - NRIIRIIIpresented by group-NHCH2CH2NH2or homologues-NH(CH2)2-4NRIVRVcan be obtained from a sulfone according to the method, according to which the diamine H2(CH2)2-4NRIVRVreacts with sulfona (connection C, Seq ID Nos. 31-45).

The reaction is carried out in the previously mentioned aprotic solvent at room temperature. Used an excess of amine is about 10 mol. The reaction is carried out in a period of time from one to several hours.

When carrying out the reaction of the appropriate amine is added to a solution of sulfone in an anhydrous aprotic solvent, and the reaction mixture was stirred at room temperature to obtain compound I (Seq ID Nos. 1-15) in which the substituent at position C-5-orn" - NRIIRIII. The target compound can be isolated by diluting the reaction mixture with an aqueous solution of TFUCK with subsequent chromatographic separation. This is suitable brasindiana column flash chromatography (C18) using 10-25% acetonitrile in water (containing 0.1% TFUC) increments of 5%. The appropriate fractions can betwen into the hydrochloride by dissolving in water and passing through a column of Bio-Rad AG2-X8 (Cl-).

If in the formula (I) R1is hydrogen, compound I' (Seq ID Nos. 1-3, 15), the nitrogen atom may be directly introduced into hemiaminal position by the formation of the azide, which is then restored in the amine, which in turn may be partially alkylated or allerban to obtain the final product.

The reaction shown in the scheme of synthesis.

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Although some natural cyclopentapeptide R1is hydrogen, more often, R1-hydroxyl. Thus, for some compounds, the compound A' scheme of synthesis obtained in the first stage from the corresponding compounds in which R1-hydroxyl.

Getting restored connection can be made by mixing appropriate hydroxycodone in solution LiClO4in diethyl ether at room temperature, adding TFUK and subsequent addition of triethylsilane. After that, the mixture is subjected to rapid stirring (4-10 h) up until the original gidroksosoedinenii no longer detected analytical HPLC. After the reaction mixture was poured into distilled water to obtain the recovered product in the form of sediment, cotesia to obtain azide in a purified or unpurified form.

The products in which R1is hydrogen, can be obtained by adding the modified cyclopentapeptide to the previously prepared solution N3, N3can be obtained from sodium azide and TFUCK. The reaction proceeds at room temperature to obtain a product containing an azide group, and which may be isolated by conventional methods and purified HPLC.

Purified azide can be restored with hydrogen on palladium deposited on charcoal, in aminosidine in a manner similar to the above.

Amines obtained, as indicated above, and having a primary amino group-NH2can then be alkylated by conventional means to obtain a substituted amino group. In short, the alkylation can be carried out by the interaction of the corresponding alkylhalogenide with amidon (compound I, NRIIRIII= NH2); Seq ID Nos. 1-15) in an aprotic solvent in the presence of a base to obtain the monosubstituted amine (compound I, NRIIRIIIwhere RII-C1-C4alkyl, C3-C4alkenyl, (CH2)2-4OH, (CH2)2-4NRIVRV). The latter can be separated from the reaction mixture by conventional methods.

AMI is icname ways to obtain a substituted amino group. It is assumed that the acyl group-CO(CH2)1-4NH2. Because alleluya acid contains a primary amino group, prior to acylation it is protected by the Deputy type benzyloxycarbonyloxy group. It is preferable to use activated ester type pentafluorophenyl. The acylation can be performed in an aprotic solvent in the presence of a base, such as di(ISO-propyl)ethylamine, at room temperature in a period of time from one to several hours to obtain the product of the acylation. The product can be isolated by diluting the reaction mixture with methanol and purified HPLC (compound I, NRIIRIII=-NHCO(CH2)1-4NH2).

Aminosidine, in which the amino group in hemiaminal position is completely replaced, i.e., in which RIIor RIIIare not hydrogen atoms,

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it is preferable to obtain from sulfone (compound B, Seq ID Nos. 31-45) interaction with the appropriate substituted amine otherIIRIII. The reaction can be carried out by adding the amine to a stirred solution of the sulfone in a period of time sufficient for reaction. The product can be isolated and cleaned the salts of related acids. The connection in the normal way its allocation is obtained as a salt with any acid (related acid). This is usually salt TFUCK. Thus obtained salt can be dissolved in water and filtered through an anion exchange column, the bearing of the target anion. The eluate containing the target salt, can be concentrated to obtain the salt in the form of a solid product.

Compounds of the present invention are active against many fungi, in particular against Candida species. Fungicidal properties can be illustrated by the minimal fungicidal concentration (MFC) specific to certain organisms, species of Candida in the experience dilution of the culture of microorganisms is performed in an environment of nitrogen fungal basics with the addition of 1% dextrose (Yeast Nitrogen Base (DIFCO) medium with 1% dextrose (YNBD).

In the present experience of connection with an initial concentration of 5 mg/ml were solubilization in 100% DMSO. The prepared solution of the drug dilution water was brought to a concentration of 512 μg/ml, so that the final concentration of DMSO was 10%. Then the solution is distributed from multiple-dosing in the first column 96-cell plates (each cell contained 0.075 ml YNBD), so that the resulting concentration of the drug is Oia medication was changed from 256 to 0.12 mcg/ml

Using spectrophotometer at 600 nm, it was found that cultures of microorganisms by the age of 4 h under test correspond to 0.5 standard Mac Farland. This suspension was diluted in YNBD in the ratio of 1:100 to achieve a cell concentration of cells 1 - 5 104forming a colony of organisms (EYE)/ml. Aliquots of the suspension (0.075 ml) were inoculated in each cell microtitre plates, which resulted in a final cell concentration of 5 - 25 103EYE/ml and a final concentration of drugs from 128 to 0.06 µg/ml Each experiment included a number of control cells that do not contain drugs, and a number of control cells that do not contain cells.

After 24 h incubation microtitre plates gently shook on the vibrating table for mixing the cell suspension. To transfer 1.5 ál samples from each cell 96-cell microtitre plates in a single cell containing Sabouraud's dextrose agar (SDA) was used microdosing MIC-2000. Grafted cells with SDA was subjected to incubation for 24 h at 35oC. the Results are shown in table. 6.

Activity of compounds against fungi in vitro can be demonstrated for the same compounds in the experiment in vivo.

Organisms from roseanniacal cells hemacytometer counting and bring to 3.75 105cells/ml and Then 0.2 ml of this suspension is injected into a vein located on the tail of a mouse, so that the final vaccine is 7.5 104cells/mouse.

Experience continue intraperitoneal (I. P.) the introduction of an aqueous solution of compound I with different concentrations twice a day over the next 4 days the female mouse DBA/2 weighing 18 to 20 g, which were previously infected with Candida albicans in the manner described previously. The control group of mice injected (I. P. ) distilled water. After 7 days mice killed by carbon dioxide, both kidneys are removed aseptically and placed in sterile plastic bags containing 5 ml of sterile saline. Kidney packages fracture to a homogeneous state, diluted with sterile saline and aliquots applied to the surface of SDA plates. Plates subjected to incubation at 35oC for 48 h, and fungal colony number for counting colony forming organisms (EYE) on 1 g of kidney. Connection(1), (2), (3) and (4) gave more than 99% the effect of reducing the concentration of secreted EYE Candida at 0.09 and 0.375 mg/kg (I. P.) twice a day for 4 consecutive days.

Compounds of the present invention is also useful for ingibirovaniem. The effectiveness of compounds that are the subject of the present invention, therapeutic or anti-infective purposes can be demonstrated in experiments on rats with depressed immunity.

In the presented experiment was determined by the efficiency of compound I-6-1 (R1= OH; R2= H; R3=CH2CH2NH2; RI=TMTD; RII=H; RIII=CH2CH2NH2). In rats Spraga-doli (Sprague-Dawley) weighing about 250 g immunity was suppressed by adding dexasone in drinking water (2.0 mg/l). Rats were kept on resemble low-protein diet for 7 weeks to induce the development of pneumocystosis pneumonia from latent infection. Before applying the medication two rats were euthanized to confirm the presence of pneumonia Pneumocystis carinii pneumonia. It was found that both rats infected. Five rats weighing about 150 g, twice a day for 4 days was administered subcutaneously (sc) compound I-6-1 in 0.25 ml of solvent (distilled water). Also conducted control experience with clean solvent. All animals continued to receive dexason in drinking water and were on resemble low-protein diet throughout the treatment period. After treatment, all animals were euthanized, and lungs were removed and surveys the research compound I-6-1 decreased the content of cysts of P. carini 5 rats at least 90% at a dose of 0.075 mg/kg and the survival of all the rats.

Outstanding properties are used most effectively if the compound is transferred to a new pharmaceutical composition comprising a suitable carrier, in accordance with generally accepted technologies make medicines.

New compositions contain at least therapeutic antifungal or antipneumocystis dose of active compounds. In General, the composition contains at least 1 wt.% connections I. Concentrated composition used for dilution may contain 90% or more. Songs include forms suitable for oral, local, parenteral (including intraperitoneal, subcutaneous, intramuscular and intravenous) administration, reception through the nose, in the form of candles and inhalation. The composition can be Packed finely mixed with components suitable as an enabling environment.

Compositions intended for oral administration may be solid or liquid. For the preparation of liquid formulations of therapeutic agent can be mixed with a liquid carrier such as water, glycols, oils, alcohols, etc. and for solid pulse, carbonates of calcium and sodium, calcium phosphate, talc, lactose, usually with a lubricant such as calcium stearate, together with the baking powder, etc. because of the ease of their reception of tablets and capsules seem to be the most promising dosed by oral forms. The most promising is the preparation of dosage forms to facilitate acceptance and uniformity of dosage. Compositions in the form of dosage forms are the subject of the present invention.

The compositions can be formulated for injection and may be in the form of suspensions, solutions or emulsions in oily or aqueous solvents, such as 0.85% sodium chloride or 5% dextrose in water, and may contain suspendida, stabilizing and/or dispersing agents. Buffering agents, as well as saline or glucose may be added to make the solution isotonic. The compounds can also be solubilisation in alcohol/propylene glycol or polyethylene glycol for intravenous drip infusion. These compositions can also be presented in a dosage form in ampoules or mnogochasovykh packages, mainly with the addition of preservative. On the other hand, the active ingredients is sirovina form" used in the description and the formula refers to physically discrete portions, each of which contains a predetermined amount of the active ingredient together with a pharmaceutical carrier, calculated to produce the desired therapeutic effect. Examples of such dosage forms are tablets, capsules, pills, powder packets, briquettes, measuring doses in capsules or mnogorazovye packaging, etc. Portions dosage in the present invention usually contains from 100 to 200 mg of one of the compounds.

If the connection is intended for antifungal use, it can be used any method of introduction. For the treatment of fungal infections is commonly used oral or intravenous administration.

If the connection should be used in the fight against pnevmaticheskie infections, it is desirable to directly enter it into the lungs or bronchi. For this reason, it is preferable inhalation. For inhalation packing of the compounds of the present invention are easy to make in the form of an aerosol spray, presents the packages under pressure or by spray. The preferred system for inhalation is a metered dose aerosol, which may be represented in the form of a suspension or solution of compound I in the stationary invention can be used as tablets, capsules, local products, inhalation powders, candles, etc., the solubility of these compounds makes them suitable for use in compositions for injection, as well as in liquid compositions, and aerosol sprays.

The following examples illustrate the invention, but shall not be construed as limitations.

Examples 1-3 illustrate the receipt of the products by the first of the above methods, namely through sulfon. This method can be used to obtain any of the connections, but should be used to achieve a good output if R1=OH.

Example 4 and further illustrating the receipt of the products by direct substitution of nitrogen for oxygen in hemiaminal position 5-orn". This method is preferred if R1, RIIand RIII-H.

Example 3 illustrates the use as starting material the compound in which R3was restored to the-CH2CH2NH2from a natural product, in which R3- CH2CONH2. Similarly, for compounds in which R3-CH2CN can also be used partially modified connection.

Examples 9 and 10 illustrate the conduct of turning ASS="ptx2">

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Part A. the production of intermediate 1-[4-hydroxy-5-(EPI)-aminoacetic-N2-(10,12-dimethyl-1-oxometalates)ornithine] -5-(3-hydroxyglutaric)-6-(3-hydroxyproline)echinocandin B(Seq ID No. 34).

A solution of 500 mg (0.47 mmol) pneumocandin Bo(Seq ID No. 19), 5.34 g (47 mmol) of the hydrochloride of 2-amino-ethandiol and 109 mg (0.47 mmol) of (1S)-(+)-10-sulfocarbamoyl acid in 40 ml of anhydrous DMF was stirred at 25oC for 6 days. The reaction mixture is diluted with 40 ml of water and flash chromatographic "LICHRO-PREP" RP18 (40-63 μm, 15.0 g), Packed in a column using 10% acetonitrile in water. Column elute 10-40% acetonitrile in water, collect two factions 120 ml at each step changes in the concentration of 10%. Of the two factions in 40% aqueous acetonitrile get 185 ml of a material, which is purified preparative HPLC "BOND" C8 (21.2 x 250 mm), elwira 40-45% acetonitrile in water with addition of 0.1% TFUK and obtain 128 mg of triptoreline 1-[4-hydroxy-5-(EPI)-aminoacetic - N2-(10,12-dimethyl-1-oxometalates)-ornithine] -5-(3 - hydroxyglutaric)-6-(3-hydroxyproline)echinocandin B in the form of a white anhydrous solids.

PMR (400 MHz, CD3OD) 1.34 (d, J=6.3 Hz, 3H), 2.89 (m, 1H), 4.72 (d, J= 4.9 Hz, 1H)

Mass spectrum (Li), m/e 1131 (MH+Li)+< / BR>
Part B. Poluchenogo as described in part A, in 15 ml of acetonitrile, diluted with water in the ratio 1:1, add "OXONE" (324 mg equivalent to 1.06 mmol acidic potassium persulfate). After 45 min the solution was diluted with the same volume of water and quickly chromatographic using brasindiana (C18) flash chromatographic column, elwira 35-43% acetonitrile in water with addition of 0.1% TFUCK with step changes in the concentration of 2%. The fractions containing the product, lyophilizers and receive 357 mg (yield 86%) EPI-sulfone.

PMR (400 MHz, CD3OD) 3.48 (m, 2H), 3.55 (m, 1H), 3.71 (m, 1H), 3.91 (dd, 1H), 4.00 (m, 1H), 5.17 (dd, 1H), 6.76 (d, 2H), 7.16 (d, 2H).

Part C. the product of formula (I); a compound 1-4 (Seq ID No. 4).

To a stirred solution of 1.2 g (0,945 mmol) EPI-sulfone, obtained as described in part B, in 20 ml of anhydrous DMF, add Ethylenediamine (568 mg, 9.45 mmol). After 1 h the analysis of the reaction mixture using HPLC (RP-C18, 40% CH3CN/H2O (0.1% TFUC) showed complete conversion into two polar product in the ratio of 37:63. For obremeniaet column (C18) flash chromatography with 10-40% acetonitrile in water with addition of 0.1% TFUK as eluent step changes in concentration of 5% followed by lyophilization of the corresponding fractions with the formation of 200 mg (yield 21%) norm is H), 4.10 (m), 5.04 (dd, J=8.7 and 3.2 Hz, 1H), 5.09 (dd, J = 8.5 and 4.2 Hz, 1H), 5.18 (br s, 1H), 6.74 (d, J = 8.6 Hz, 2H), 7.12 (d, J=8.6 Hz, 2H), 7.47 (d, J = 8,6 Hz, 1H), 7.71 (d, J = 10.0 Hz, 1H), 8.11 (d, J = 8.7 Hz, 1H), 8.71 (d, J = 8.7 Hz, 1H).

Mass spectrum (Li), m/e 1113.5 (MLi)+< / BR>
(Bis)-triptorelin obtained previously is dissolved in water and the solution passed through a column of Bio-Rad AG2-X8 (Cl-) wash water. The eluate containing the product, lyophilizers with the formation of the above compounds in the form of (bis)-hydrochloride.

Lyophilization of the fractions containing base material, gives an EPI-product.

PMR (400 MHz, CD3OD) / 3.02 (m, 1H), 3.14 (m, 3H), 4.16 (m, 1H), 5.10 (dd, 1H), 6.76 (d, 2H), 7.14, (d, 2H).

Mass spectrum (Li), m/e 1113.9 (MLi)+.

Example 2.

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Part A. the intermediate sulfone (Seq ID No. 36).

Starting material, the compound A-6RI=DMTT (Seq ID No. 21), receive as described for these compounds in the section "Obtaining source materials."

Connection A-6 then turn in EPI-tizaidine B-6(Seq ID No. 36) in a manner analogous to that described in part A of example 1.

To a stirred solution of 285 mg (0.241 mmol) of compound B-6 in 14 ml of acetonitrile, diluted with water in the ratio 1:1, add "OXONE" (162 mg equivalent 0.530 mmol sour persulfates column (C18) flash chromatography with 30 to 45% acetonitrile in water with addition of 0.1% TFUK as eluent step changes in concentration of 5% followed by lyophilization containing the product fractions with the formation of 212 mg EPI-sulfone (compound C-6 to Seq ID No. 36).

Yield 84%.

PMR (400 MHz, CD3OD) 3.08 (m, 2H), 3.46 (t, J = 6.6 Hz, 2H), 3.68 (m), 5.05 (m), 6.77 (d, J = 8.5 Hz, 2H), 7.15 (d, 8.5 Hz, 2H).

Mass spectrum (Li), m/e 1039.9.

Part B. the product of the formula (2) (compound 1-6; RII=H; RIII=2 aminoethyl); Seq ID No. 6.

To a stirred solution of compound C-6, obtained as described in part A (418 mg, 0.305 mmol) in 10 ml of anhydrous DMF, add Ethylenediamine (183 mg, 3.05 mmol). After 1 h the analysis of the reaction mixture using HPLC (RP-C18, 35% CH3CN/H2O (0.1% TFUC) showed complete conversion into two polar product in the ratio of 36:64. The reaction mixture is diluted with a solution containing 190 ml of water and 0.4 ml TFUK, and chromatographic. For obremeniaet column (C18) flash chromatography with 10% to 25% acetonitrile in water with addition of 0.1% TFUK as eluent step changes in concentration of 5% followed by lyophilization of the corresponding fractions with the formation of 111 mg of the product in the form of (Tris)-trifenatate. The output of 21%.

PMR (400 MHz, CD3OD) 1.17 (J=6.2 Hz), 2.44 (dd, J = 7.0 and 13.2 Hz, 1H), 4.10 (m), 2.7-3.0 (m, 4H), 3.06 (t, J = 7.0 Hz, 2H), 3.82 (m, 3H), 3.97 (dd, J = 11.2 and 3.2 Hz, 1H), 4.03 (m, 2H), 4.70 (d, J = 2.3 Hz, 1H), 5.00 (d, J = 3.3 Hz, 1H), 6.75 in (d, J = 8,6 Hz, 2H), 7.11 (d, J = 8.6 Hz, 2H).

Mass spectrum (Li), m/e 1099,9 (MLi)+.

(Tris)-Triforce the DOI. The eluate containing the product, lyophilizer education 93 mg of the above compound in the form of (Tris)-hydrochloride.

Example 3.

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A. Obtaining azide (Seq ID No. 49).

To a stirred solution of 297 mg (0,257 mmol) of apiculture (example 1, part B) in 10 ml of anhydrous DMF added lithium azide (126 mg, 257 mmol). After 1 h the analysis of the reaction mixture using HPLC (RP-C18, 40% CH3CN/H2O (0.1% TFUC) showed complete conversion in a single, significantly less polar product in the ratio of 37:63. For obremeniaet column (C18) flash chromatography with 30-65% acetonitrile in water as eluent step changes in concentration of 5% followed by lyophilization of the corresponding fractions with the formation of the crude azide. Using preparative HPLC (C18, 40-45% CH3CN/H2O (0.1% TFUC) with step changes in concentration of 5% obtained azide compound D-4 (Seq ID No. 49).

PMR (400 MHz, CD3OD) to 1.14 (d, J = 6,1 Hz, 3H), 2,50 (dd, J = 15.6 and a 9.9 Hz, 1H), 2,84 (dd, J = 15.6 and 3,3 Hz, 1H), 3,95 (dd, J = 11.2 and a 3.1 Hz, 1H), of 4.05 (m, 2H), 4,56 (m, 3H), to 4.98 (dd, J = 8.5 and a 3.5 Hz, 1H), 5,10 (dd, J = 8.3 and 4.2 Hz, 1H), of 5.26 (dd, J = 8,5 and 2.2 Hz, 1H), 6,74 (d, J = 8,6 Hz, 2H), 7,12 (d, J = 8,6 Hz, 2H), 7,44 (d, J = 8,3 Hz, 1H), 7,76 (d, J = a 9.9 Hz, 1H), compared to 8.26 (d, J = 8,1 Hz, 1H), 8,83 (d, J = 8,7 Hz, 1H), 9,00 (d, J = 8,5 Hz, 1H).

IR spectrum (Nujol), 2110 cm-1.

is, 0,126 mmol) and 10% Pd/C (137 mg) in glacial acetic acid (10 ml) saturated with hydrogen under a pressure equal to the pressure in the supply tank, for 14 hours, the Catalyst was removed by filtration, and the filtrate lyophilizer to obtain the crude amine. Purification using preparative HPLC (C18, 35-41% CH3CN/H2O (0.1% TFUC) with step changes in the concentration of 3%), followed by lyophilization of the appropriate fractions gives Aza compound I-1, RII, RIII= H (Seq ID No. 1) in the form of triptoreline.

Yield 48%.

PMR (400 MHz, CD3OD) of 1.13 (d, J = 6,1 Hz, 3H), 2.49 USD (dd, J = 15.6 and is 9.8 Hz, 1H), 2,81 (dd, J = 15.6 and 3,4 Hz, 1H), 3,97 (dd, J = 11.1 and a 3.1 Hz, 1H), a 4.03 (m, 1H), 4,11 (m, 1H), 4,47 (dd, J = 11.7 and a 5.5 Hz, 1H), 4,57 (m, 2H), 5,00 (m, 1H), 5,10 (m, 1H), 5,14 (d, J = 2,2 Hz, 1H), 6,74 (d, J = 8,6 Hz, 2H), 7,12 (d, J = 8,6 Hz, 2H), 7,42 (d, J = 8,3 Hz, 1H), 8,89 (d, J = 8,8 Hz, 1H).

Mass spectrum m/z 1071,0 (MLi)+.

Triptorelin dissolved in water and the solution passed through a column of Bio-Rad AG2-X8 (Cl-) wash water. The eluate containing the product, lyophilizer education 66 mg of compound I-4, RII, RIII= H in the form of hydrochloride.

In the following experiments, the solvent A is 95% water, 5% acetonitrile, 0.1% TFUK, and solvent B was 95% acetonitrile, 5% water, 0.1% TFUCK. Used the expression "in a vacuum" and "rotor" refers to udaleniya azide. Compound D (Seq ID no 46).

Pneumocandin B0(compound A-4; Seq ID No. 19) (5,00 g, 4,69 mmol) dissolved in 2 m LiClO4in diethyl ether at room temperature. Add with stirring TFOC (2,50 ml), then the solution is added triethylsilane (5,00 ml). A heterogeneous mixture is intensively stirred for 6 h, after which analytical HPLC (C18 "BOND", 45% solvent A, 55% solvent B, 0.1% TFUK, 1.5 ml/min) did not reveal the presence of the original pneumocandin B0in quantities of more than trace. The mixture is then poured into 200 ml of distilled water, filtered and dried in air. The wet solid product was stirred with diethyl ether, filtered and dried in air; obtain 5.6 g of crude monomolekulyarnogo pneumocandin B0(compound A-1, Seq ID No. 16).

Selected crude product in solid form is added to a pre-prepared solution HN3obtained by dissolving cooling NaN3(a 3.06 g, and 47.0 mmol) in 100 ml TFUCK. After stirring at room temperature for 30 min, the reaction mixture was poured into 350 ml of distilled water and stirred for 15 minutes the Precipitate is filtered off, dissolved in methanol and the solvent is distilled off in a rotary evaporator. Estate to remove volatile impurities. The mixture is purified in two equal portions using preparative HPLC (C18 "DELTAPAK", 60 ml/min, fraction of 48 ml) using varying concentrations of eluent from 70% A/ 30% B to 50% A/ 50% b Corresponding fractions (determined by UV absorption at = 220/ and 277 nm) are combined. Insufficiently purified fractions also combine and the process is repeated as described above. In this way we obtain 1.78 g (yield 35%) of azide-D-1 (Seq ID no 46).

PMR (400 MHz, CD3OD) 7,02 (d, 2H), 6,69 (d, 2H), and 5.30 (d, 1H), 5,11 (d, 1H), to 4.98 (d, 1H), 2,74 (dd, 1H), 1,13 (d, 3H).

Mass spectrum (Li), m/z 1081 (MH+Li)+.

B. Obtaining an amine of the formula (4). Compound I-1 (RII, RIII= H; Seq ID No. 1).

Purified azide (1.50 g), the compound D-1 obtained as described above, is dissolved in 40 ml of methanol. Add 33% aqueous acetic acid (15 ml), and then 0.20 g of 10% Pd/C, then purge the reaction vessel with nitrogen. Replacing the nitrogen with hydrogen and intensively stirred mixture in an atmosphere of hydrogen for 3 hours, the Suspension is filtered through a glass filter (dthen= 0.2 μm) and a clear solution is evaporated to dryness on a rotary evaporator. The residue is dissolved in about 20 ml of distilled water, and raids!! lyophilizer. Obtain 1.47 g (95%) of the target amino compounds (Seq ID No. 1) in the form of white">

Mass spectrum (Li), m/z 1055 (MH+Li)+.

Example 5.

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A. Obtaining an intermediate benzyloxycarbonyl connection (Seq ID No. 1).

In 1 ml of DMF is dissolved amine of the formula (4) (example 4) (200 mg, 0.180 mmol) and pentafluorophenyl N-benzyloxycarbonyl-3-aminopropanal. Add di(ISO-propyl)ethylamine (0.035 ml, 198 mmol) and stirred the mixture at room temperature for 1 h, the Reaction mixture was diluted with 2 ml of methanol and purified preparative HPLC (C18 "DELTAPAK", faction 48 ml) with varying concentrations of eluent from 70% A/30% B to 48% A/52% B. the Appropriate fractions (determined by UV absorption at = 220 and 277 nm) are combined, and raids!! lyophilizer. Get 100 mg (44%) of the target intermediate product.

PMR (400 MHz, CD3OD) 7.32 (d, 5H), 7.01 (d, 2H), 6.99 (d, 2H), 5.64 (bd, 1H), 1.18 (d, 3H).

Mass spectrum (Li), m/z 1259 (MLi)+.

B. Obtaining 3-aminopropanol the compounds of formula (5);

Compound I-1 RII=H, RIII=CO(CH2)2NH2(Seq ID No. 1).

Benzyloxycarbonyl connection (see part A) (94 mg, 0.075 mmol) dissolved in a mixture of 3 ml of methanol, 1 ml water and 0.2 ml of acetic acid. Add 10% Pd/C (48 mg) and purge the reaction vessel with nitrogen. Then blow sexest in a rotary evaporator obtain a solid product. The product is dissolved, about 4 ml of 50% aqueous acetonitrile, and raids!! lyophilizer. Receive 80 mg (91%) of target compound of the formula (5) in the form of a white solid.

PMR (400 MHz, CD3OD) 7.01 (d, 2H), 6.69 (d, 2H), 6.67 (d, 1H), 5.10 (d, 1H), 4.99 (d, 1H), 3,12 (m, 2H), 1.91 a (s, 3H), 1.17 (d, 3H).

Mass spectrum (Li), m/z 1125 (MLi)+.

Example 6.

< / BR>
Obtaining N-methylimidazolidine formula (6); compound I-1 (RII=H; RIII=CH3) (Seq ID No. 1)

Amine of formula (5) example 5 (45.6 mg, 0.135 mmol) was dissolved in 0.5 ml of dry DMF. Add 0.021 ml (0.473 mmol) iodomethane, then 0.0824 ml (0.473 mmol) of di(ISO-propyl)ethylamine. After 24 hours stirring at room temperature distillation on a rotary evaporator to remove volatile matter and the crude product is analyzed mass spectrometrically.

Mass spectrum (Li), m/z 1068 (MLi)+.

Example 7.

< / BR>
A. Obtaining an intermediate nitrile (N-cyanomethylene) of compound I-1; RII=H; RIII=CH2CN (Seq ID No. 1).

Aminosidine, obtained as described in example 4 (4540 mg, 0.451 mmol), dissolved in 3 ml of dry DMF. Add 0.063 ml (0.902 mmol) bromoacetonitrile, which was pre-purified by passing through nebuloni the mixture is stirred for 12 h, then diluted with a small volume of water. The solution is purified preparative HPLC (C18 "DELTAPAK", the change of concentration from 70% A/30%B to 47% A/ 53%B, fraction of 48 ml). The appropriate fractions, as determined by UV absorption at 220 and 277 nm, collect, and raids!! lyophilizer. Get 338 mg (62%) of the target intermediate cyanomethylene connection in the form of water-insoluble solids.

PMR (400 MHz, CD3OD) 7.01 (d, 2H), 6.69 (d, 2 H), 5.12 (dd, 1H), 5.01 (dd, 1H) 3,80 (s, 2H), 2.76 (dd, 1H), 1.15 (d, 3H).

Mass spectrum (Li), m/z 1094 (MH+Li)+.

B. Obtaining N-aminoacylase the compounds of formula (7).

Compound I-1; RII=H; RIII=(CH2)2NH2(Seq ID No. 1).

Nitrile (cyanomethylene) the compound obtained above (300 mg, 0.249 mmol), dissolved in 5 ml of methanol, then add 237 mg (0.997 mmol) of uranyl chloride Nickel (II). Three portions are added to a solution of 189 mg (4.99 mmol) of sodium borohydride. Immediately formed black precipitate, after which the mixture is stirred for 15 min at room temperature. A heterogeneous mixture is diluted with approximately 20-40 ml of water and add about 10-15 ml of 2 N. HCl. Stirring is continued for another 45 min before until a black precipitate will not dissolve with the formation of blue-green is the 48 ml). The appropriate fractions, as determined by UV absorption at 220 and 277 nm, collect, and raids!! lyophilizer. Receive 180 mg (55%) of the desired product. The substance is dissolved in 30 ml of water and passed through an ion exchange column (in the form of Cl-), washed with distilled water. The solution raids!! and lyophilizers. Obtain 149 mg (recovery 94%) target aminoacylase the compounds of formula (7) in the form of a white solid.

PMR (400 MHz, CD3OD) 7.01 (d, 2H), 6/69 (d, 2H), 5.11 (dd, 1H), 5.07 (dd, 1H), 1.14 (d, 3H).

Mass spectrum (Li), m/z 1098 (MH+Li)+.

Example 8.

< / BR>
A. Obtaining an intermediate azide. (Seq ID N 47).

Nitrile pneumocandin Bo(2.00 g, 1.91 mmol) is dissolved in 24 ml of 2M solution LiClO4in diethyl ether. Add 2.00 ml of triethylsilane, then 1 ml TFUK, the mixture is intensively stirred for 6 h at room temperature. The mixture is then poured into 300 ml of water, stirred and filtered. Substance from the filter is dissolved in minimum quantity of methanol, and the solvent is distilled off on a rotary evaporator. Residual water is removed by azeotropic distillation with 100% ethanol, and the residue left overnight in a high vacuum to remove volatile impurities. Get monoposto is) placed in a round bottom flask, equipped with a stirrer and placed in a cooling bath. Slowly add 50 ml TFUK, the cooling bath removed and the mixture is stirred for 2 hours the Mixture was poured into 300 ml of water and filtered. The solid is dissolved in methanol, the solvent is distilled off on a rotary evaporator and kept in a high vacuum to remove volatile impurities. The crude material is purified preparative HPLC (C18"DELTAPAK", the change of concentration from 55% A/45% B to 45%a/b, fraction of 56 ml). The appropriate fractions, as determined by UV absorption at 220 and 277 nm, collect, and raids!! dipylidium. Obtain 0.59 g (29%) target intermediate azide.

PMR (400 MHz, CD3OD) 7.00 (d, 2H), 6.69 (d, 2H), 5.34 (d, 1H), 5.07 (dd, 1H), 5.00 (m, 1H), 2.88 (dd, 1H), 1.17 (d, 3H).

Mass spectrum (Li), m/z 1036 (M-N2+Li)+.

C. Obtaining the compounds of formula (8) (Seq ID No. 48).

Purified azide (part a)(0.15 g, 0.142 mmol) dissolved in a mixture of 4 ml of methanol, 1 ml water and 0.5 ml of acetic acid. To the solution was added 50 mg of 10% Pd/C. the Reaction vessel is rinsed with N2then H2. The mixture is intensively stirred for 5 h at room temperature under pressure of H21 ATM. Subsequent filtration through a glass filter (dthen=0.2 μm) and removal of volatiles on a rotary COI is PMR (400 MHz, CD3OD) 7.00 (d, 2H) 6.69 (d, 2H), 5.04 (d, 1H), 5.01 (m, 1H), 2.79 (dd, 1H), 1.18 (d, 3H).

Mass spectrum (Li), m/z 1037 (MH+Li)+.

Example 9.

< / BR>
Obtaining the amino compounds of the formula (9) (Seq ID No. 3).

The purified azide-nitrile from example 8 (part a) (44 mg, 0.0416 mmol) dissolved in 1.5 ml of methanol, then add CoCl2H2O (59 mg, 0.25 mmol). Then portions carefully added NaBH4(8 x 12 mg, 2.50 mmol). Black heterogeneous reaction mixture is stirred for 30 min at room temperature. The reaction is stopped by adding approximately 1.5 ml of 2 N. HCl and sufficient to dissolve the precipitate amounts of acetic acid. The cloudy solution was diluted with 3 ml of water and purified preparative HPLC (C18 "BOND", the change of concentration from 70% A/30% to 60%a/40% B, 15 ml/min, fraction 15 ml). The appropriate fractions, as determined by UV absorption at 210 and 277 nm, collect, and raids!! lyophilizer. Get 38 g (72%) of the compounds of formula (9) in the form of a white solid.

PMR (400 MHz, CD3OD) 6.99 (d, 2H), 6.70 (d, 2H), 5.11 (d, 1H), 5.0 (m, 2H), 1.17 (d, 3H).

Mass spectrum (Li), m/z 1041 (MH+Li)+.

Example 10.

< / BR>
A. Obtaining an intermediate bis-nitrile compound (compound I-2, RII=H; RIII=CH2CN; RIohoho DMF. Add 0.064 ml (0.917 mmol) bromoacetonitrile, which was pre-purified by passing through a small layer of MgSO4- NaHCO3, then add di(ISO-propyl)ethylamine (0.155 ml, 0.917 mmol). The reaction mixture is stirred for 18 hours the Mixture is diluted with water and purified preparative HPLC (C18 "DELTAPAK", 60 ml/min, the change in concentration from 70% A/30%to 50%, fraction of 48 ml). The appropriate fractions, as determined by UV absorption at 220 and 277 nm, collect, and raids!! lyophilizer. Obtain 198 mg (36%) of target compound I-1; RII=H; RIII= CH2CN.

PMR (400 MHz, CD3OD) 7.00 (d,2H), 6.69 (d, 2H), 5.08 (dd, 1H), 5.01 (dd, 1H), 3.73 (s, 2H), 2.79 (dd, 1H), 1.18 (d, 3H).

Mass spectrum (Li) m/z 1076 (MH+Li)+.

C. Obtaining the compounds of formula (10) (SeQ ID No. 3).

Bis-nitrile (part a) (184 mg, 0.155 mmol) was dissolved in 3 ml of methanol. Also dissolved in methanol 148 mg (0.621 mmol) NiCl26H2O, and three portions add 117 mg (3.1 mmol) NaBH4. After 5 minutes add 148 mg (0.621 mmol) CoCl2H2O and stirred for further 1 min Optional add another 117 mg NaBH4and continue mixing for another 15 minutes Past 60 mg NaBH4added to bring the reaction to completion. SMEs (C18 "BOND", 15 ml/min, the change in concentration from 70% A/30% to 55% A/ 45% B, fraction 22.5 ml, 220, 277 nm) after lyophilization gives solid. The solid is dissolved in water and passed through an ion exchange column (in the form of Cl-), and raids!! lyophilizer. Get 81.1 mg (44%) of target compound of formula (10) (connection 1 - 3; Seq ID No. 3) in the form of a white solid.

PMR (400 MHz, CD3OD) 7.00 (d, 2H), 6.70 (d, 2H), 3 to 3.3 (m, 6H), 1.18 (d, 3H).

Mass spectrum (Li), m/z 1084 (MH + Li)+.

Examples 11 to 14.

In action similar to that described in example 4, the corresponding natural or modified natural cyclopeptide products obtained as described further in the preparation of starting materials, when performing separate operations is dissolved in the solution LiClO4in diethyl ether, and with stirring add there TFUK and triethylsilane for 5 to 10 hours and Then poured the mixture into water, filtered and stirred solid product with diethyl ether, dried in air and get cyclopeptide, in which R1restored to H.

Monovoltine connection add cooling to the previously obtained from NaN3and TFUK solution HN

Azide is treated with hydrogen, as described previously, using Pb/C as a catalyst, and distinguish the product from the filtrate after separation of the catalyst.

The products obtained in this way are given in table. 7.

Examples 15 to 17.

In action similar to that described in the dreamer 7, the compounds of examples 11, 13 and 14 are dissolved in DMF and added to a solution of the purified bromoacetonitrile, then di(isopropyl)amine, and the mixture is stirred for 12 - 18 h to obtain the nitrile (N-cyanomethylene) connection. In further purified preparative HPLC.

Nitrile is dissolved in methanol and chemically restored with the use of Nickel chloride (II) and sodium borohydride to obtain aminoacylase substituted compounds. The compounds obtained are given in table. 8.

Examples 18 to 21.

As a result of operations performed similar to that described in examples 1, 2 and 3, can be obtained from compounds containing listed in the table. 9 deputies.

Examples 22 to 25.

As a result of operations performed similar to that described in example 1 are obtained the following compounds (see table. 10).

Example 27.

< / BR>
The above compound is obtained by way similar to that described in example 26, substituting piperidine dimethylamine to obtain a connection in M m 1374.

Example 28.

1000 compressed tablets each containing 500 mg of the compounds of formula (2) [connection 1 - 6 (RII=H; RIII=2-amino-ethyl); Seq ID No. 6] obtained from the following ingredients:

Connection - Mass, g

Connection example 2 - 500

Starch - 750

Hydrate dibasic calcium phosphate 5000

Calcium stearate - 2.5

Micronized ingredients mix well and granularit with 10% starch paste. The granules are dried and pressed into tablets.

Example 29.

1000 hard gelatin capsules, each containing 500 mg of the same compound obtained from the following ingredients:

Connection - Mass, g

Connection example 2 - 500

Starch - 250

Lactose - 750

Talc - 250

Calcium stearate - 10

A homogeneous mixture of ingredients obtained by the mixture used for filling hard gelatin capsules consisting of two parts.


Connection example 2 - 24 mg

Lecithin, NF liquid concentrate - 1.2 mg

Trichloro(fluorine)methane, NF - 4.026 g

Dichloro(debtor)methane, NF - 12.15 g

Example 31.

250 ml solution for injection can be obtained using conventional methods from the following ingredients:

Connection - Mass, g

Dextrose - 12.5 g

Water - 250 ml

The compound of example 4 400 mg

The ingredients are mixed and then sterilized for use.

Obtaining raw materials.

A-4 if RI=GMTD, can be produced by cultivating Zalerion arboricola ADS 206868 in a nutrient medium with the use of mannitol as the primary carbon source, as described in U.S. Pat. USA N 5, 021, 341 dated 4 June 1991

A-7 if RI=GMTD, can be produced by cultivating Zalerion arboricola ADS 20868 in a nutrient medium as described in U.S. Pat. USA N 4, 931, 352 dated 5 June 1990

A-10 if RI- linoleyl, can be produced by culturing Aspergillus nidulans NRRL 11440 in nutrient medium as described in U.S. Pat. USA N 4, 288, 549 dated 8 September 1981

A-11 if RI-11-methyldecyl, can be produced by culturing AEM cultivating Zalerion arboricola ADS 20958 11440 in a nutrient medium, as described in the application ser. N 07/630, 457, filed on December 19, 1990 (Atty Docket N 18268).

Compounds in which R1is hydrogen, can be obtained as described in example 4, part A.

Compounds in which R3-CH2CN, such as A-2, A-5 and A-8, can be produced by reaction of compounds having a carboxyl group in a suitable position with an excess of cyanuric chloride in an aprotic solvent. This may be a molecular sieve. Upon completion of the reaction strainers, if used, is removed, and the filtrate is concentrated to obtain a nitrile compound, as is more fully described in the application ser. N 936, 434 dated September 3, 1992

Compounds in which R3-CH2CH2NH2such as A-3, A-6 and A-9, can be produced by chemical or catalytic reduction of the nitrile. Most suitable for this purpose is carrying out the reaction using a large molar excess of sodium borohydride with cobalt chloride, as is more fully described in the application ser. N 936, 558 dated September 3, 1992

Raw materials, in which RI- different groups of occurring in natural products, can be obtained by diallylammonium lip is the ed until until full diallylamine. These enzymes previously obtained by cultivation of microorganisms of the family Pseudomondaceae or Actinoplanaceae as described in Experentia 34, 1670 (1978) or U.S. Pat. USA 4, 293, 482, allocate describeany cyclopeptide and then acelerou describeany cyclopeptide, mixing with a suitable active ester RICOX to obtain A connection with the desired acyl group.

1. The cyclic peptides of the formula I

< / BR>
where R1= H or OH;

R2= H;

R3= CH2CN, CH2CH2NH2or CH2CONH2;

RI= (C9- C21)-alkyl;

RII= H, (C1- C4)-alkyl;

RIII= H, (CH2)nNH2;

n = 2 or 3,

or their additive salt of the acid.

2. Connection on p. 1, where R1= OH, R3= CH2CONH2, RI= 9,11-dimethylthiazol, RII= H and RIII= CH2CH2NH2.

3. Connection on p. 1, where R1= OH, R3- CH2CH2NH2, RI= 9,11-dimethylthiazol, RII= H and RIII= CH2CH2NH2.

4. Connection on p. 1, where R1= OH, R3= CH2CONH2, RI= 9,11-dimethylthiazol, RIIand RIII, RIIand RIII= H.

6. Connection on p. 1, where R11= H, R3= CH2CONH2, RI= 9,11-dimethylthiazol, RII= H, R3= CH2CH2NH2.

7. Connection on p. 1, where R1= H, R3= CH2CH2NH2, RI= 9,11-dimethylthiazol, RIIand RIII= H.

8. Connection on p. 1, where R1= H, R3= CH2CH2NH2, RI= 9,11-dimethylthiazol, RII= H and RIII= CH2CH2NH2.

9. Composition exhibiting antifungal and antipneumococcal activity, comprising a therapeutic dose of a compound under item 1 in a mixture with a pharmaceutically acceptable carrier.

10. A method of treating fungal infections, characterized in that it is administered to a mammal in need of such treatment, a connection on p. 1 in the amount of 100 to 200 mg per single dose.

11. The connection formulas



 

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FIELD: medicine, cardiology.

SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.

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3 ex

FIELD: medicine, phthisiology, anesthesiology.

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EFFECT: higher efficiency of compensation.

1 ex, 1 tbl

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