Glycine-substituted thieno[2,3-d]pyrimidines with combined lh- and fsh-agonistic activity

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention describes glycine-substituted thieno[2,3-D]-pyrimidines of the formula (I): wherein X represents oxygen atom (O) or H,H; A represents sulfur atom (S), -NH, -N(R6), O or a bond; R1 represents (C1-C4)-alkyl, (C2-C4)-alkenyl, unsubstituted or substituted phenyl, thienyl, pyridyl; R2 represents hydrogen atom (H), (C1-C4)-alkyl, (C1-C4)-alkoxy-(C2-C4)-alkyl or hydroxy-(C2-C4)-alkyl; R3 and R4 are chosen independently from H, (C1-C4)-alkyl and hydroxy-(C1-C4)-alkyl; R5 represents H or (C1-C4)-alkyl, and R6 represents (C1-C4)-alkyl. Compound possess agonistic activity with respect to glycoprotein hormone, in particular, to compounds possessing agonist activity with respect to luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Also, invention describes pharmaceutical compositions containing such compounds and using these compounds in medicinal therapy, in particular, for fertilization control.

EFFECT: valuable biological and medicinal properties of compounds.

11 cl, 1 tbl, 27 ex

 

The invention relates to compounds having an agonistic activity against glycoprotein hormone, in particular to the compounds with agonistic activity against luteinizing hormone (LH) and follicle-stimulating hormone (FSH). The invention also relates to pharmaceutical compositions containing such compounds and to the use of these compounds in drug therapy, in particular for controlling fertilization.

The gonadotropins play an important role in various body functions, including metabolism, temperature regulation and reproductive process. Hypofyse the gonadotropins FSH and LH, for example, play an essential role in promoting the development and maturation of the follicle, while LH is included in the induction process of ovulation (Sharp, R. Clin. Endocrinol33:787-807, 1990; Dorrington and Armstrong, Recent Prog. Horm. Res35: 301-342, 1979; Levy et al, Human Reproduction15:2258-2265, 2000).

Currently, LH will apply in the clinical setting, in combination with FSH to stimulate the ovaries, i.e. hyperstimulatory ovaries for fertilizationin vitro(IVF) and induction of ovulation in infertile, unable to ovulation women (Insler, V., Int. J. Fertility33:85-97, 1988, Navot and Rosenwaks, J. Vitro Fert. Embryo Transfer5:3-13, 1988), as well as for male hypogonadism and male infertility.

The gonadotropins act on specific t the p cells, related to sexual gland, to initiate ovarian and testicular differentiation and steroidogeneza. These pituitary and placental hormones mediated by specific receptors of the plasma membrane that are members of a large family coupled receptors G-proteins. They consist of a single polypeptide with seven transmembrane domains and are able to interact with G-proteins, leading to activation of adenylcyclase.

The gonadotropins, intended for therapeutic purposes, can be isolated from human urine and have a low purity (Morse et al, Amer. J. Reproduct. Immunol. and Microbiology17:143, 1988). Alternatively, you can get them in the form of recombinant gonadotropins. In addition to these proteins gonadotropin receptors can be activated or deactivated by using low-molecular compounds obtained by synthesis. In WO 00/61586 described bicyclic heteroaromatic compounds. Experimentsin vitroandin vivohave shown that they are useful as LH agonists.

In normal women, the release of pituitary LH and FSH is characterized by a large spike in the middle of the cycle that precedes ovulation. Ovulation is characterized by three distinctive physiological characteristics, namely maturation of oocytes, follicular rupture and what teensarea. While the role of a great surge of LH in the induction of these phenomenain vivodiscusses the role of FSH surge is less clear. However, recently it has been shown that FSH induces oocyte maturationin vitroby inducing accumulation of cells producing factor, positively overcoming induced gipoksantin stop meiosis (Lu et al, Mol. Cell. Endocrinol.164:191-196, 2000). I believe that this factor is a meiosis activating Sterol (MAS).

This invention relates to low molecular weight compounds, which have LH activity. In addition to LH activity they also exhibit FSH activity, which was unexpected. Basically, these connections represent thieno[2,3-d]pyrimidines which are substituted in position 4 of the pyrimidine ring of the phenyl group, which, in turn, substituted in the meta position. This includes Deputy triatomic spacer elements group (NH-C(O)-CH2), which, in addition, functionalized substituted amino group. Compounds, mainly, have various degrees of FSH agonistic activity, but as a rule, smaller than LH agonistic activity.

The present invention relates to glycine-substituted derivatives of thieno[2,3-d]pyrimidine of the General formula I

or their pharmaceutically acceptable the th salt, where

X represents O or H,H;

And represents S, NH, N(R6), O or a bond;

R1represents (1-4C)alkyl, (2-4C)alkenyl, phenyl or (2-5C)heteroaryl, where the phenyl or heteroaryl ring optionally substituted by one or more substituents groups: hydroxy, halogen, nitro, trifluoromethyl, cyano, amino or (1-4C)(di)alkylamino;

R2represents H, (1-4C)alkyl, (1-4C)alkoxy(2-4C)alkyl or hydroxy(2-4C)alkyl;

R3and R4independently selected from H, (1-4C)alkyl and hydroxy(1-4C)alkyl;

R5represents H or (1-4C)alkyl, and

R6can be selected from the same groups as described for R1.

The term (1-4C)alkyl as used in the definition of formula (I), means a branched or unbranched alkyl group containing 1-4 carbon atoms, for example methyl, ethyl, n-drank, isopropyl, n-butyl, sec-butyl or tert-butyl.

The term (1-4C)alkenyl, as used in the definition of formula (I), means a branched or unbranched alkenylphenol group containing 2-4 carbon atoms, for example vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 2-methyl-2-propenyl, 1-methyl-1-propenyl, 1-ethyl vinyl.

The term (1-4C)alkoxy(1-4C)alkyl means an alkyl group containing 1-4 carbon atoms linked through an atom of color is Yes with another alkyl group, containing 1-4 carbon atoms, with alkyl groups have the same meanings as defined above.

The term (1-4C)alkoxy(2-4C)alkyl means an alkyl group containing 1-4 carbon atoms linked through an oxygen atom with another alkyl group containing 2-4 carbon atoms, with alkyl groups have the same meanings as defined above.

The term hydroxy(1-4C)alkyl means a hydroxyl group attached to an alkyl group containing 1-4 carbon atoms, with alkyl groups have the same meanings as defined above.

The term hydroxy(2-4C)alkyl means a hydroxyl group attached to an alkyl group containing 2-4 carbon atoms, with alkyl groups have the same meanings as defined above.

The term (1-4C)(di)alkylamino means one or two alkyl groups containing 1-4 carbon atoms, as defined above, attached to the nitrogen atom.

The term (2-5C)heteroaryl means optionally substituted aromatic group containing 2-5 carbon atoms, comprising at least one heteroatom selected from N, O and/or S, such as imidazolyl, thienyl, furyl or pyridyl.

The term halogen means fluorine, chlorine, bromine or iodine.

It was found that the compounds of the above formula I demonstrate agonistic LH and FSH activity. In bioanalysisin vitro, in which the used cells SNO, stable tropicabana receptor LH or FSH person, respectively, were determined value EU50in relation to the LH receptor is less than 5·10-8M, while in respect of the FSH receptor is EU50was less than 10-5M Usually FSH activity is from about 1% from the stimulation of LH agonist to about 10% of the stimulation of LH agonist.

The present invention also relates to pharmaceutical compositions comprising a derivative thieno[2,3-d]pyrimidine or its salt of the General formula I.

Thus, the compounds of the present invention can be used in therapy. Another aspect of the present invention relates to the use of thieno[2,3-d]pyrimidine of the General formula I to obtain drugs for control of fertilization, more preferably induce ovulation. Compounds of the present invention is used for the activation of both LH and FSH receptors. In this regard, the compounds of the present invention can be used in a method for the treatment of women with problems related to fertilization.

Connection thieno[2,3-d]pyrimidine according to the present invention can contain one or more chiral carbon atoms. In this regard, the compounds can be obtained in the form of chiral pure compounds or as mixtures of diastereomers and/or enantiomers. Methods of obtaining hyral is but pure compounds are well known in the prior art, for example, crystallization or chromatography.

For use in the treatment of salts of compounds of formula I are salts in which the counterion is pharmaceutically acceptable. However, the acid additive salts of bases of formula I can also find application, for example, in obtaining or purification of pharmaceutically acceptable compounds. All salts are pharmaceutically acceptable or not included in the scope of this invention.

Examples of the acid additive salts include salts derived from mineral acids such as hydrochloric acid, phosphoric acid, sulfuric acid, preferably hydrochloric acid, and organic acids such as citric acid, tartaric acid, acetic acid, lactic acid, maleic acid, malonic acid, fumaric acid, glycolic acid, succinic acid, etc.

Suitable routes of administration of compounds of the formula I or their pharmaceutically acceptable salts, also denoted herein as the active ingredient include intramuscular injection, subcutaneous injection, intravenous injection or intraperitoneal injection, oral and intranasal administration. Preferably the compounds can be administered orally. The exact dose and mode of administration of the active ingredient, or a pharmaceutical composition will be for the Iset, mainly from the expected therapeutic effect (treatment of infertility; contraception) and can vary depending on the particular compound, the route of administration and the age and condition of the particular subject, which is injected the drug.

Mainly parenteral administration requires lower doses than other methods of administration, which are more dependent on adsorption. However, the dosage for people preferably contains 0,0001-25 mg per kg of body weight. The desired dose may be presented as a single dose or as multiple subds introduced at suitable intervals during the day. If the recipient is a woman, the dose can be entered via the appropriate day intervals during the menstrual cycle follicular support or as a single dose to induce ovulation. Dose, as well as the mode of administration may vary, for recipients, male and female.

In the case of applicationsin vitroorex vivofor example in IVF use, the compounds of the present invention is used in the incubation medium at a concentration of about 0.01 to 5 mg/ml

The present invention thus also relates to pharmaceutical compositions comprising a compound of the thieno[2,3-d]pyrimidine of the formula I, in a mixture with pharmaceutically acceptable excipients is mi and optionally, other therapeutic agents. Excipients can be "acceptable" in terms of their compatibility with other ingredients of the composition and safety for recipients.

Pharmaceutical compositions include compositions suitable for oral, rectal, nasal, local (including dermal, buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. The composition can be obtained by any method well known in the field of pharmacy, for example, using the methods described in Gennaro et al., Remington,'s Pharmaceutical Sciences (18thed., Mack Publishing company, 1990, see, in particular, part 8:Pharmaceutical Preparations and Their Manufacture).

These include the stage of linking the active ingredient with any auxiliary agent. Auxiliary(e) agent(agents), also called incremental ingredients include agents traditionally used in the art (Gennaro, supra), such as fillers, binders, diluents, disintegrating agents, lubricants, dyes, fragrances and moisturizing agents.

Pharmaceutical compositions suitable for oral administration, are presented as separate dosage units such as pills, tablets or capsules, or in the form of powder or granules, or in view of the solution or suspension. The active ingredient may also be presented as a bolus or paste. The composition can also be obtained in the form of suppositories or enemas for rectal administration.

For parenteral administration suitable compositions include aqueous and non-aqueous sterile injection. The composition can be provided in containers that includes one or more doses, such as sealed vials and ampoules, and can be kept under the conditions of freeze drying (in dried form), when you need only the addition of sterile aqueous media such as water, before use.

Compositions or formulations suitable for administration by nasal inhalation, include finely ground powders or dust particles generated by metered-dose pressurized aerosol devices, sprays or insufflators.

Connection thieno[2,3-d]pyrimidine according to the present invention can also be administered in the form of implantable pharmaceutical devices, consisting of a core of active substance enclosed in a membrane that regulates the speed of its release. Such implants are used subcutaneously or topically, and the release of the active ingredient occurs with approximately constant speed for a relatively long time, for example from weeks to several years. With the person receiving implantable pharmaceutical devices as such, known in the art, for example as described in European patent 0303306 (AKZO N.V.).

Thus, compounds according to this invention, can be used for the same clinical purposes as natural LH, with the advantage that they possess FSH activity, exhibit improved properties stability and can be entered in different ways.

Compounds of the present invention represented by the General formula (I)receive, mainly, by nucleophilic substitution of compounds of General formula (II), where Q=Cl or Br, amines of General formula (III) in a suitable solvent such as N,N-dimethylformamide or THF, at room temperature in the presence of a tertiary base, such as N,N-diisopropylethylamine. Most of the amines of General formula (III) are commercially available.

Derivatives of the formula (II), where Q=Cl or Br, you can get the regioselective acylation meta-aniline derivatives of the formula (V-a) anhydrides of carboxylic acids of type (IV), where Q=Cl or Br, in the presence of a tertiary base, such as N,N-diisopropylethylamine, in a suitable solvent such as dichloromethane or THF.

The compounds of formula (V) can be obtained by means well known in the field of recovery functionalevolutionary in derivatives of the formula (VI), using a suitable reducing agent, such as hydrogen, in the presence of a metal catalyst (Pd/Pt). Appropriate recovery methods described in: P.M. Carabateas, P.R. Brundage, K.O. Gellote, M.D. Gruett, R.R. Lorenz, J. Heterocycl. Chem.21, 1849 (1984). Alternatively, the recovery can be carried out by tin chloride (II) in proton solvent such as ethanol, in the presence of hydrochloric acid at elevated temperature (J. Heilbron, J. Chem. Soc, 1279 (1940)).

Thienopyrimidine General formula (VI) can be obtained by condensation of the carboxylic acid (VII) with tert-butylamine under the influence of the binding agent, such as tetrafluoroborate O-(benzotriazol-1-yl)-N,N,N,N,-tetramethylurea (TBTU) or hexaphosphate patrimonialization (PyBrOP), and a tertiary base, for example N,N-diisopropylethylamine.

Saponification of the corresponding complex ethyl ester (VIII) to the carboxylic acid (VII) in the presence of a base such as lithium hydroxide, potassium hydroxide or sodium hydroxide in aqueous dioxane at elevated temperature (80°C to the boiling point).

Bicyclic system General formula (VIII) is obtained by substitution of the chlorides of the formula (X) ethylmercaptan the action of N,N-diisopropylethylamine with the respective base catalyzed the cyclization of the intermediate thioesters (IX). This method of forming rings thieno[2,3-d]pyrimidine described in: S.A. Abdel-Hady, M.A. Badawy, Y.A. Ibrahim, Sulfur Lett.9, 101 (1989) and S. Tumkevicius, Liebigs Ann., 1703 (1995).

Suitable conditions for the reaction of cyclization are ethoxide sodium in ethanol or N,N-diisopropylethylamine in toluene/ethanol (1/1, V/V), at the boiling point.

The compounds of formula (X) can be synthesized following the methods described in the literature, as, for example, described A.A. Santilli, D.H. Kim and S.V. Wanser, J. Heterocycl. Chem.8, 445, 1971. In a typical experimental conditions amide having the General structure (XI), was treated with POCl3at elevated temperature (80°C to the boiling point). The addition of an appropriate solvent, for example dioxane, and/or adding or PCl5or N,N-dimethylaniline to the reaction mixture may reduce reaction time and to ensure higher yields chloride (X).

The main path leading to the lactam of formula (XI)comprises the condensation of ethylcyanoacrylate with 3-nitrobenzaldehyde and compounds (XII), which can be estimatedin (XII-a), smokeview (XII-b), monosubstituted guanidine (XII-c), disubstituted guanidine (XII-d) or amidine (XII-e).

In a typical experiment, these shall cyanoacetate, 3-nitrobenzaldehyde and derivative (XII) is suspended in a suitable solvent, for example ethanol, methanol, N,N-dimethylformamide, N-methylpyrrolidinone, tetrahydrofuran or pyridine, and add a base such as potassium carbonate, sodium acetate, sodium methoxide or ethoxide sodium. The reaction takes place at elevated temperature (from 70°C to the boiling point). After filtration residues are transferred into water and acidified (pH 2), then there is precipitation products (XI) (S. Kambe, K. Saito and H. Kishi, Synthesis, 287 (1979); A.M. Abd-Elfattah, S.M. Hussain and A.M. El-Reedy, Tetrahedron39, 3197 (1983); S.M. Hussain, A. A. El-Barbary and S.A. Mansour, J. Heterocycl. Chem.22, 169 (1985)).

Alternatively, the compounds of this invention, in which A=N, is represented by the formula (I-a), can be obtained from derivatives of the sulfoxide of General formula (XIII) by nucleophilic substitution with amine nucleophiles of General structure (XIV). The reaction is usually carried out at elevated temperature in the presence of a tertiary base, such as N,N-diisopropylethylamine, in a suitable solvent such as 1,4-dioxane.

Similarly, the compounds of this invention, in which A=O is represented by formula (I-b), can be obtained from derivatives of the sulfoxide of General formula (XIII) by nucleophilic substitution by nucleophiles alkoxide of the General structure (XV). The reactions is carried out in the presence of tert-butoxide potassium with excess alcohol R 1-OH.

Derivatives sulfoxide of General formula (XIII) can be obtained by oxidation of compounds of General formula (I), where R1=IU and A=S, represented by formula (I-c). The oxidation is carried out with the help of 3-chloroperbenzoic acid in triperoxonane acid. The acidic nature of the solvent prevents the oxidation of 5-amino groups into the corresponding 5-nitrosopropane.

Well known methods of determining binding of the receptor, as well as analyses of thein vitroandin vivoto determine the biological activity of gonadotropins. Typically, the downregulation of the receptor is in contact with the test compound, and measuring the binding, or stimulation or inhibition of a functional response.

In order to measure the functional response, the selected DNA encoding a gene of receptor LH or FSH, preferably the receptor of the person, expressed in suitable host cells. Such a cell can be a cell of the ovary of the Chinese hamster, but also suitable are other cells. Preferred cells are cells derived from mammals (Jia et al, Mol.Endocrin., 5:759-776, 1991).

The methods of constructing cell lines expressing recombinant LH or FSH, is well known in the art (Sambrook et al., Molecular Cloning: a Laboratory Manual, Cold Spring Harbo Laboratory Press, Cold Spring Harbor, latest edition). The receptor expression is achieved by the expression of DNA that encodes a desired protein. Currently in the art well-known methods site-directed mutagenesis, ligation added sequences, PCR and construction of suitable expression systems. Part or full design DNA encoding the desired protein can be obtained by synthetic means, using standard methods of solid-phase synthesis, it is preferable to include restriction sites to facilitate ligation. For coding DNA sequences may be provided with suitable control elements for transcription and translation included the coding sequence. As is well known, at the present time, there are expression systems that are compatible with a wide range of hosts, including prokaryotic hosts, such as bacteria, and eukaryotic hosts such as yeast, plant cells, insect cells, mammalian cells, bird cages, etc.

Cells expressing the receptor is then contacted with a test compound to observe binding, or stimulation or inhibition of a functional response.

Alternatively, in order to measure the binding connection, you can use a dedicated cell membranes containing Express the integration of the receptor.

To measure binding, you can use radioactively or fluorescently labeled compounds. The control connection can be used recombinant human LH or FSH. Alternatively also conduct analyses of competitive binding.

Other analysis includes screening compounds to determine the properties of the agonist of LH or FSH receptor by determining the stimulation of receptor-mediated camp accumulation. Thus, this method involves receptor expression on the cell surface of the host cell and the effect on the cell under test compounds. Then measure the amount of camp. The level of camp must be low or high, depending on inhibitory or stimulating effect of the test compound when it is binding with the receptor.

In addition to direct measurement, for example, levels of camp in exposed to compounds in the cell, you can use cells, which in addition to transfection encoding the receptor DNA, are also transfitsirovannykh second DNA that encodes a reporter gene, expression of which corresponds to the level of camp. Such reporter genes may be induced by camp or can be designed in such a way that they are associated with new, responsive to the camp elements. Mainly, the expression of rap is Chernogo gene can be controlled by any sensor element, reacting to changes in the levels of camp. Suitable reporter genes include LacZ, alkaline phosphatase, Firefly luciferase and green fluorescent protein. The principles of such TRANS-activation assays well known in this field and are described, for example, Stratowa, Ch, Himmler, and A Czernilofsky, A.P. (1995) Curr.Opin.Biotechnol.6:574.

For selection of compounds exhibiting activity against LH or FSH receptor, tested at a concentration of 10-5M should show the activity that constitutes more than 20% of maximum activity when LH or FSH is used as a control for comparison. Another criterion may be the largest EU50that should be <10-5M, preferably <10-7M

The specialist in this area should be clear that the desired size EU50depend on the tested compounds. For example, the connection, EC50which is less than 10-5M, generally considered to be a candidate for the choice of drug. Preferably this value is less than 10-7M. However, the connection having a higher value EU50but which is selective with respect to specific receptor may be an even better candidate.

For screening compounds which are agonists LH receptor, you can also use the bioanalysis of Leydig cells m the Chi (Van Damme, M., Robersen, D. and Diczfalusy, E. (1974). Acta Endocrinol. 77: 655-671 Mannaerts, B., Kloosterboer, H. and Schuurs, A. (1987). Neuroendocrinology of reproduction. R. Rolland et al. Eds., Elsevier Science Publishers B.V., 49-58). In this analysis the stimulation of LH receptor-mediated production of testosterone can be measured in Leydig cells isolated from male mouse.

FSH agonistic activity of the compounds may be defined on the model ofex vivousing cultured murine follicles, in accordance with Nayudu, P. and Osborn, S. (1992, J. Reproduction and Fertility 95:349-362). This mouse ovarian follicles were isolated and cultured in the presence of compounds which are agonists of FSH to induce growth of follicles. Measure the diameter of follicles and estradiol in the culture medium was the growth of the follicle.

To measurein vivoactivity of compounds against LH investigated the induction of ovulation in immature female mice. In this analysis of immature female mice were primiraly isolated from urine FSH and about 48 hours were treated with compound is an agonist of LH. After treatment with agonist LH animals were killed and using microscopic analysis estimated the number of female germ cells in the fallopian tube.

To measurein vivoactivity of compounds against FSH immature female rats at time 0, 8, 24 and 32 hours were treated with compound which is an agonist of FSH, for inductionof follicle. After 52 hours after the start of the experiment, animals were injected with hCG by injection to induce ovulation. Animals were killed 72 hours after the start of the experiment and using microscopic analysis estimated the number of female germ cells in the fallopian tube. In addition, we determined the weight of the ovary.

The compounds of this invention can be applied in a clinical setting, in the same mode as currently used LH or hCG. Such applications include LH replacement in subjects with decreased function of sexual glands, and male and female, introduction to the middle of the cycle to induce ovulation (ovulation induction (OI) or controlled ovarian hyperstimulation (SON) or stimulation of the corpus luteum).

The following examples are to illustrate the invention and in no way should be interpreted as limiting the scope of invention.

EXAMPLES

Example 1

tert-Butyl 5-amino-2-methylthio-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) 5-Cyano-4-(3-nitrophenyl)-2-methylthio-6-hydroxypyrimidine

A mixture of sulfate S-methylisothiazoline (69,0 g), 3-nitrobenzaldehyde (75,0 g), ethylcyanoacrylate (56,0 ml) and potassium carbonate (72.5 g) in absolute EtOH (1500 ml) was stirred at 60°C for 16 hours. The reaction mixture was cooled to 0°in the bath with ice. Received wasp is OK was filtered, washed with absolute EtOH and dissolved in hot water (100°). The solution was cooled to room temperature, acidified 2 N. HCl to pH 2 and was cooled to 0°in the bath with ice. The precipitate was filtered and washed with ice water. Remaining in the sediment water was removed by joint evaporation with 1,4-dioxane.

Output: 54,0,

MS-ESI: [M+H]+=289,0.

TLC: Rf=0,3, silica gel, DCM/MeOH=9/1 (V/V).

(b) 6-Chloro-5-cyano-4-(3-nitrophenyl)-2-methylthiopyrimidin

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-methylthio-6-hydroxypyrimidine (example 1(a)of 25.0 g) in dry 1,4-dioxane (300 ml) was added POCl3(100 ml). After 3 hours at 90°the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane (100 ml) and the resulting solution was cooled to 0°C. was Carefully added to ice water. The precipitate was filtered and washed with water. Remaining in the sediment water was removed by joint evaporation with 1,4-dioxane.

Output: 26,0,

MS-ESI: [M+H]+=307,0.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(C) Ethyl 5-cyano-4-(3-nitrophenyl)-2-methylthio-6-(ethoxycarbonylmethyl)pyrimidine

To a stirred solution of ethyl 2-mercaptoacetate (9,3 ml) and 6-chloro-5-cyano-4-(3-nitrophenyl)-2-methylthiopyrimidine (example 1(b), 26,0 g) in a mixture of EtOH (250 ml) and DCM (250 ml) was added DIPEA (15.7 ml). Chere is C 1 hour at room temperature, to the mixture was added 0.1 G. aqueous HCl (500 ml) and then was extracted with DCM (3x500 ml), dried (MgSO4) and concentrated under reduced pressure.

Output: 28,0,

MS-ESI: [M+H]+=391,4.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(d) Ethyl 5-amino-4-(3-nitrophenyl)-2-methylthieno[2,3-d]pyrimidine-6-carboxylate

A mixture of ethyl 5-cyano-4-(3-nitrophenyl)-2-methylthio-6-(ethoxycarbonylmethyl)pyrimidine (example 1(C)of 28.0 g) and DIPEA (30 ml) in a mixture of toluene (150 ml) and EtOH (150 ml) was stirred at the boiling point under reflux (100° (C) for 16 hours. Then the mixture was cooled to room temperature and concentrated under reduced pressure. The remaining DIPEA were joint removed by evaporation with toluene.

Output: 28,0,

MS-ESI: [M+H]+=391,4.

TLC: Rfor =0.6, silica gel, heptane/EtOAc=3/2 (V/V).

(e) Ethyl 5-amino-4-(3-nitrophenyl)-2-methylthieno[2,3-d]pyrimidine-6-carboxylate

To a mixture of ethyl 5-amino-4-(3-nitrophenyl)-2-methylthieno[2,3-d]pyrimidine-6-carboxylate (example 1(d)of 28.0 g), concentrated aqueous HCl (15 ml) and tin chloride (II) (41,0 g) in 1,4-dioxane (400 ml) was added EtOH (400 ml). The mixture was stirred at 90°C for 16 hours. Then the mixture was cooled to room temperature and concentrated under reduced pressure. The residue is suspended in EtOAc (1000 ml). To achieve pH 10-11) was added 4 N. aqueous NaOH. The mixture was thoroughly stirred and the organic is the second layer was separated, dried (MgSO4) and concentrated under reduced pressure.

Output: 21,0,

MS-ESI: [M+H]+=361,0.

TLC: Rfor =0.6, silica gel, heptane/EtOAc=3/2 (V/V).

(f) 5-Amino-4-(3-AMINOPHENYL)-2-methylthieno[2,3-d]pyrimidine-6-carboxylic acid

To a solution of ethyl 5-amino-4-(3-nitrophenyl)-2-methylthieno[2,3-d]pyrimidine-6-carboxylate (example 1(e), or 21.0 g) in a mixture of 1,4-dioxane (300 ml) and water (100 ml) was added potassium hydroxide (32,4 g). After 16 hours at 90°the mixture was cooled to 10°and with thorough stirring, was added 2 N. aqueous solution of citric acid (300 ml). The precipitate was filtered off, washed with water (180 ml) and dried in vacuum.

Yield: 14.0 g

MS-ESI: [M+H]+=333,0.

TLC: Rf=0,5, silica gel, DCM/MeOH=9/1 (V/V).

(g) tert-Butyl 5-amino-4-(3-AMINOPHENYL)-2-methylthieno[2,3-d]pyrimidine-6-carboxamide

To a solution of 5-amino-4-(3-AMINOPHENYL)-2-methylthieno[2,3-d]pyrimidine-6-carboxylic acid (example 1(f), 14.0 g), DIPEA (17,4 ml) and tert-butylamine (7,3 g) in DCM/DMF (1/1, V/V, 250 ml) was added TBTU (16,1 g). After 3 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(3x100 ml), 0.1 G. of aqueous HCl solution (100 ml) and water (100 ml). The organic layer was concentrated under reduced pressure. The crude product was purified by crystallization from warm absolute EtOH (300 ml).

Output: 10.5V,

MS-ESI: [M+H]+/sup> =388,2.

HPLC: Rf=30,72 min, Luna C-18(2), 5 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: water/ACN/MeOH=90/9,5/0,5 to 0/95/5, cycle time=50 minutes

(h) tert-Butyl 5-amino-2-methylthio-4-(3-(2-bromoacetamide)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-2-methylthio 4-(3-AMINOPHENYL)thieno[2,3-d]pyrimidine-6-carboxamide (example 1(g), 1.08 g) and DIPEA (2,43 ml) in dry DCM (20 ml) was added bromocatechol (615 mg). After 3 hours at room temperature the mixture was diluted with DCM, washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified column chromatography on silica gel, using as eluent heptane/EtOAc=3/2 (V/V).

Output: 910 mg.

MS-ESI: [M+H]+=510,2.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(i) tert-Butyl 5-amino-2-methylthio-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-methylthieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) in DCM (5 ml) was added N-methyl-2-aminoethanol (250 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product acidalia, using column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 112 mg (g TPUK).

MS-ESI: [M+H]+=503,2.

HPLC: Rt=of 11.45 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 2

tert-Butyl 5-amino-2-methylthio-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) in DCM (5 ml) was added 2-amino-2-methylpropanol (250 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 67 mg (g TPUK).

MS-ESI: [M+H]+=from 517.2.

HPLC: Rt=12,67 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, t is mperature furnace=40° C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 3

tert-Butyl 5-amino-2-methylthio-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) and N,N-diisopropylethylamine (0,20 ml) in DCM (5 ml) were added hydrochloride licensedialog ester (200 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Output: 133 mg (g TPUK).

MS-ESI: [M+H]+=from 517.2.

HPLC: Rt=11.87 per min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 4

tert-Butyl 5-amino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-2-methylthio-4-(3-(2-b is macedonica)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 200 mg) in DCM (5 ml) was added N-(2-methoxyethyl)ethylamine (266 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 84 mg (g TPUK).

MS-ESI: [M+H]+=531,2.

HPLC: Rt=br12.62 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 5

tert-Butyl 5-amino-4-(3-((N-(R-1-methoxycarbonyl-2-methylprop-1-yl)glycinyl)amino)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-2-methylthio-4-(3-(2-bromoacetamide)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) and N,N-diisopropylethylamine (0,20 ml) in DCM (5 ml) was added the hydrochloride of the methyl ester of D-valine (250 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified at the same time HPLC, using column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 77 mg (g TPUK).

MS-ESI: [M+H]+=559,2.

HPLC: Rt=13,22 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 6

tert-Butyl 5-amino-4-(3-((N,N-bis-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-2-methylthio-4-(3-(2-bromoacetamide)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) in DCM (5 ml) was added N,N-bis-(2-methoxyethyl)amine (400 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 166 mg (g TPUK).

MS-ESI: [M+H]+=561,3.

HPLC: Rt=13,62 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, tempera is ur furnace=40° C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 7

tert-Butyl 5-amino-2-methylthio-4-(3-((2,3-dihydroxypropyl-1-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxamide (example 1(h), 250 mg) in DCM (5 ml) was added 3-amino-2-hydroxypropane (250 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Output: 164 mg (g TPUK).

MS-ESI: [M+H]+=519,2.

HPLC: Rt=br12.62 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 8

tert-Butyl 5-amino-2-methylthio-4-(3-((1,3-dihydroxypropyl-2-yl)glycinyl)amino)phenyl-thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-methylthio-thieno[2,3-d]pyrimidine-6-carboxy the IDA (example 1(h), 250 mg) in DCM (5 ml) was added 2-amino-3-hydroxypropane (250 mg). After 16 hours at room temperature the mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 117 mg (g TPUK).

MS-ESI: [M+H]+=519,2.

HPLC: Rt=br12.62 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 9

tert-Butyl 5-amino-2-phenyl-4-(3-((N-ethyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) 5-Cyano-4-(3-nitrophenyl)-2-phenyl-6-hydroxypyrimidine

A mixture of benzenedimethanamine (16.4 g), 3-nitrobenzaldehyde (15.1 g), ethylcyanoacrylate (11.2 ml) and potassium carbonate (16.6 mg) in absolute EtOH (250 ml) was stirred at 60°C for 8 hours. The reaction mixture was cooled to 0°in the bath with ice. The precipitate was filtered off, washed with absolute EtOH and heated in water (100°) until then, until he got a clear solution. The solution of ohlord is whether the up to 50° C, acidified to pH 2 by adding 2 N. aqueous solution of HCl and cooled to 0°in the bath with ice. The precipitate was filtered and washed with water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 15,0,

MS-ESI: [M+H]+=spreads for about 319.2.

TLC: Rf=0,3, silica gel, DCM/MeOH=9/1 (V/V).

(b) 6-Chloro-5-cyano-4-(3-nitrophenyl)-2-phenylpyrimidine

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-phenyl-6-hydroxypyrimidine (example 9(a), 15.0 g) and dimethylaniline (0.5 ml) in dry 1,4-dioxane R.A. (200 ml) was added POCl3(50 ml). After 3 hours at 90°With the warm mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane was added ice-cold water. The precipitate was filtered and washed with water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 15,8,

MS-ESI: [M+H]+=337,4.

TLC: Rf=0.8, silica gel, heptane/EtOAc=3/2 (V/V).

(C) Ethyl 5-cyano-4-(3-nitrophenyl)-2-phenyl-6-(ethoxycarbonylmethyl)-pyrimidine

To a stirred solution of ethyl 2-mercaptoacetate (5,15 ml) and 6-chloro-5-cyano-4-(3-nitrophenyl)-2-phenylpyrimidine (example 9(b), to 15.8 g) in a mixture of EtOH (125 ml) and DCM (125 ml) under nitrogen atmosphere was added DIPEA (8,71 ml). After 2 hours at room temperature the mixture was diluted with DCM until dissolved, washed with 0.5 N. aqueous HCl solution, dried (MgSO4) is concentrated under reduced pressure.

Output: 19,7,

MS-ESI: [M+H]+=UAH 421,2.

TLC: Rf=0,7, silica gel, heptane/EtOAc=3/2 (V/V).

(d) Ethyl 5-amino-4-(3-nitrophenyl)-2-phenylthieno[2,3-d]pyrimidine-6-carboxylate

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-phenyl-6-(ethoxycarbonylmethyl)pyrimidine (example 9(C)of 19.7 g) in a mixture of absolute EtOH (100 ml) and toluene R.A. (100 ml) was added DIPEA (20,0 ml). After 48 hours at 100°the mixture was cooled to 0°C. the precipitate was filtered off, washed with cold EtOH and dried in vacuum at 40°C.

Output: 17,0,

MS-ESI: [M+H]+=UAH 421,2.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(e) Ethyl 5-amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxylate

To a solution of ethyl 5-amino-4-(3-nitrophenyl)-2-phenyl-thieno[2,3-d]pyrimidine-6-carboxylate (example 9(d), 16.6 g) in 1,4-dioxane R.A. (250 ml) was added tin chloride (II) (23,0 g) in absolute EtOH. Was added 37% aqueous HCl solution (6.9 ml) and the mixture is boiled under reflux for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The residue is suspended in EtOAc (500 ml). To achieve pH 10-11) was added 4 N. aqueous NaOH. The mixture was diluted by addition of a saturated aqueous solution of NaCl. The organic layer was separated, dried (MgSO4) and concentrated under reduced pressure.

Output: 17,0,

MS-ESI: [M+H]+=UAH 421,2.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(f) 5-Amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxylic acid

To a solution of ethyl 5-amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxylate (example 9(e), 17.0 g) in a mixture of 1,4-dioxane (210 ml) and water (80 ml) was added potassium hydroxide (20,0 g). After 16 hours at 90°the mixture was cooled to 0°C. the precipitate was filtered, suspended in water (300 ml) and cooled to 0°C. the Mixture was acidified to pH 3 by adding 2 N. aqueous solution of citric acid and was stirred at 0°until reaching room temperature for 2 hours. The precipitate was filtered off, washed with water and dried in vacuum at 40°C.

Output: 13,3,

MS-ESI: [M+H]+=363,0.

TLC: Rf=0,2, silica gel, DCM/MeOH=95/5 (V/V).

(g) tert-Butyl 5-amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxamide

To a mixture of 5-amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxylic acid (example 9(f), 13.3 g) in a mixture of DCM (250 ml) and DMF (250 ml), under nitrogen atmosphere, was added DIPEA (15.3 ml), tert-butylamine (9,3 ml) and TBTU (14.1 g). After 3 hours at room temperature the mixture was diluted with DCM and washed with saturated aqueous NaHCO3, 0.1 N. aqueous solution of HCl and saturated aqueous NaCl. The organic layer was dried (MgSO4) and concentrated under reduced pressure. The crude product is cleaned is whether using chromatography on silica gel, using as eluent heptane/EtOAc=3/7 to 1/1 (V/V).

Output: 14,7,

MS-ESI: [M+H]+=418,4.

TLC: Rf=0,4, silica gel, heptane/EtOAc=3/2 (V/V).

(h) tert-Butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-phenylthieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-AMINOPHENYL)-2-phenylthieno[2,3-d]pyrimidine-6-carboxamide (example 9(g), 5.8 g) and DIPEA (12,2 ml) in DCM (50 ml) was added dropwise bromocatechol (2,80 ml). After 3 hours at room temperature the mixture was diluted with DCM, washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel, using as eluent heptane/EtOAc=3/2 (V/V). Received a mixture of 1:1 (mol/mol) tert-butyl 5-amino-4-(3-(2-bromoacetamide)-phenyl)-2-phenyl-thieno[2,3-d]pyrimidine-6-carboxamide and tert-butyl 5-amino-4-(3-(2-chloroacetamido)-phenyl)-2-phenyl-thieno[2,3-d]pyrimidine-6-carboxamide.

Output: 2,6,

MS-ESI: [M+H]+=540,2, [M,+H]+=494,2.

TLC: Rf=0,3, silica gel, heptane/EtOAc=3/2 (V/V).

(i) tert-Butyl 5-amino-2-phenyl-4-(3-((N-ethyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-phenylthieno[2,3-d]pyrimidine-6-carboxamide (example 9(h), 500 mg) in DCM (5 ml) was added N-ethyl-2-aminoethanol (500 mg). Poluparallelnye for 17 hours at room temperature the reaction mixture was diluted with DCM (100 ml), washed with saturated aqueous NaHCO3(1 M, h ml), dried (MgSO4) and concentrated in vacuum. The crude product was purified HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Output: 271 mg (g TPUK).

MS-ESI: [M+H]+=547,2.

HPLC: Rt=11,88 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 10

tert-Butyl 5-amino-2-phenyl-4-(3-(N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-phenylthieno[2,3-d]pyrimidine-6-carboxamide (example 9(h), 500 mg) and N,N-diisopropylethylamine (DIPEA, 1 ml) in DCM (5 ml) were added hydrochloride methyl ester of glycine (700 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (100 ml), washed with saturated aqueous NaHCO3(1 M, h ml), dried (MgSO4) and concentrated in vacuum. The crude product was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) is 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous solution TFUK and water.

Output: 321 mg (g TPUK).

MS-ESI: [M+H]+=547,2.

HPLC: Rt=12,54 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 11

tert-Butyl 5-amino-2-(2-furyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) 2-amidinopropane

In chilled (bath with ice, 0° (C) the reaction vessel containing 2-furonitrile (13 ml), was added a cooled (0° (C) and saturated ethanolic HCl solution (40 ml). The resulting solution was left to reach room temperature and was stirred under nitrogen atmosphere for 48 hours. After concentrating the reaction mixture under vacuum, the residue containing the corresponding 2-forretirement, re-dissolved in ethanol (20 ml) and stirred at 0°C in nitrogen atmosphere. Then was added a saturated ethanolic ammonia solution (40 ml) and the reaction mixture was stirred in a sealed reaction vessel for 48 hours. After filtering the reaction mixture, the filtrate was concentrated under reduced pressure. The crude compound was used in the next stage without additional purification.

O is d: 15,0,

(b) 5-Cyano-4-(3-nitrophenyl)-2-(2-furyl)-6-hydroxypyrimidine

A mixture of 2-amidinopropane (example 11(a), 15 g), 3-nitrobenzaldehyde (24 g), ethylcyanoacrylate (17 ml) and potassium carbonate (25 g) in absolute EtOH (300 ml) was stirred at 60°C for 16 hours. The reaction mixture was cooled to 0°in the bath with ice. The precipitate was filtered off, washed with absolute EtOH and heated in water (100° (C) to obtain a milky white suspension. The suspension was cooled to 50°C, acidified to pH 2 by adding 2 N. aqueous solution of HCl, and cooled to 0°in the bath with ice. The precipitate was filtered and washed with ice water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 16,0,

MS-ESI: [M+H]+=309,2.

TLC: Rf=0,3, silica gel, DCM/MeOH=9/1 (V/V).

(C) 6-Chloro-5-cyano-4-(3-nitrophenyl)-2-(2-furyl)pyrimidine

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-(2-furyl)-6-hydroxypyrimidine (example 11(b), 16.0 g) and dimethylaniline (0.5 ml) in dry 1,4-dioxane R.A. (250 ml) was added POCl3(50 ml). After 2 hours at 90°With the warm mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane and added to ice water. The precipitate was filtered and washed with water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 16,0,

MS-ESI: [M+H]+/sup> =327,2.

TLC: Rf=0.75, silica gel, heptane/EtOAc=3/2 (V/V).

(d) Ethyl 5-cyano-4-(3-nitrophenyl)-2-(2-furyl)-6-(ethoxycarbonylmethyl)pyrimidine

To a stirred solution of ethyl 2-mercaptoacetate (5,4 ml) and 6-chloro-5-cyano-4-(3-nitrophenyl)-2-(2-furyl)pyrimidine (example 11(C), 16.0 g) in a mixture of EtOH (125 ml) and DCM (125 ml) under nitrogen atmosphere was added DIPEA (9.1 ml). After 2 hours at room temperature the mixture was diluted with DCM until dissolved, washed with 0.5 N. aqueous HCl solution, dried (MgSO4) and concentrated under reduced pressure.

Output: 20,0,

MS-ESI: [M+H]+=411,2.

TLC: Rf=0,7, silica gel, heptane/EtOAc=3/2 (V/V).

(e) Ethyl 5-amino-4-(3-nitrophenyl)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a stirred solution of ethyl 5-cyano-4-(3-nitrophenyl)-2-(2-furyl)-6-(ethoxycarbonylmethyl)pyrimidine (example 11(d), 20 g) in a mixture of absolute EtOH (100 ml) and toluene R.A. (100 ml) was added DIPEA (20,0 ml). After 48 hours at 100°the mixture was cooled to 0°C. the precipitate was filtered off, washed with cold EtOH and dried in vacuum at 40°C.

Output: 20,

MS-ESI: [M+H]+=411,2.

TLC: Rfor =0.6, silica gel, heptane/EtOAc=3/2 (V/V).

(f) Ethyl 5-amino-4-(3-AMINOPHENYL)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a solution of ethyl 5-amino-4-(3-nitrophenyl)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 11(e), 20 g) in 1,4-dioxane R.A. (250 ml)was added a solution of tin chloride (II) (28,0 g) in absolute EtOH (250 ml). Was added 37% aqueous solution of HCl (8.5 ml) and the mixture was heated under reflux (90° (C) for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The residue is suspended in EtOAc (500 ml). To achieve pH 10-11) was added 4 N. aqueous solution of NaOH. The mixture was diluted by addition of a saturated aqueous solution of NaCl. The organic layer was separated, dried (MgSO4) and concentrated under reduced pressure.

Output: 17,5,

MS-ESI: [M+H]+=381,2.

TLC: Rf=0,4, silica gel, heptane/EtOAc=3/2 (V/V).

(g) 5-Amino-4-(3-nitrophenyl)-2-(2-furyl)thieno[2,3-d] pyrimidine-6-carboxylic acid

To a solution of ethyl 5-amino-4-(3-AMINOPHENYL)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 11(f)of 17.5 g) in a mixture of 1,4-dioxane (210 ml) and water (80 ml) was added potassium hydroxide (23,0 g). After 8 hours at 90°the mixture was cooled to 0°C. the precipitate was filtered, suspended in water (300 ml) and cooled to 0°C. the Mixture was acidified to pH 3 by adding 2 N. aqueous citric acid solution, and stirred at 0°C to room temperature for 2 hours. The precipitate was filtered off, washed with water and dried in vacuum at 40°C.

Output: 16,9,

MS-ESI: [M+H]+=353,2.

TLC: Rf=0,2, silica gel, DCM/MeOH=95/5 (V/V).

(h) tert-Butyl 5-amino-4-(3-AMINOPHENYL)-2-(2-furyl)thieno[2,3-d]Piri is one-6-carboxamide

To a solution of 5-amino-4-(3-AMINOPHENYL)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxylic acid (example 11(g)and 16.9 g) in a mixture of DCM (250 ml) and DMF (50 ml) under nitrogen atmosphere was added DIPEA (19.2 ml), tert-butylamine (to 11.6 ml) and TBTU (17,7 g). After 3 hours at room temperature formed a significant amount of yellow precipitate, which was filtered. The residue was washed with diethyl ether and dried in vacuum at 40°C.

Output: 18,0,

MS-ESI: [M+H]+=408,2.

TLC: Rf=0,4, silica gel, heptane/EtOAc=3/2 (V/V).

(i) tert-Butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-AMINOPHENYL)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 11(h), 250 mg) and DIPEA (0.5 ml) in DCM (5 ml) was added bromocatechol (100 l). After 3 hours at room temperature the mixture was diluted with DCM, washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel, using as eluent heptane/EtOAc=3/2 (V/V). Received a mixture of 1:1 (mol/mol) tert-butyl 5-amino-4-(3-(2-bromoacetamide)-phenyl)-2-(2-thienyl)-thieno[2,3-d]pyrimidine-6-carboxamide and tert-butyl 5-amino-4-(3-(2-chloroacetamido)-phenyl)-2-(2-thienyl)-thieno[2,3-d]pyrimidine-6-carboxamide.

Yield: 124 mg

MS-ESI: [M+H]+=540,2, [M,+H]+=494,2.

TLC: Rf=0,3, silicagel, heptane/EtOAc=3/2 (V/V).

(j) tert-Butyl 5-amino-2-(2-furyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)-phenyl)-2-(2-furyl)-thieno[2,3-d]pyrimidine-6-carboxamide (example 11(i), 130 mg) in DCM (5 ml) was added N-methyl-2-aminoethanol (200 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, 2x10 ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 81 mg (g TPUK)

MS-ESI: [M+H]+=523,2

HPLC: Rt=br11.01 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 12

tert-Butyl 5-amino-2-(2-furyl)-4-(3-(N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 11(i), 130 mg) in DCM (5 ml) was added 2-and the Ino-2-methylpropanol (260 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, h ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 54 mg (g TPUK).

MS-ESI: [M+H]+=537,2.

HPLC: Rt=of 11.15 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 13

tert-Butyl 5-amino-2-(2-furyl)-4-(3-(N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-furyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 11(i), 130 mg) and DIPEA (0.5 ml) in DCM (5 ml) were added hydrochloride methyl ester of glycine (324 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, 2x10 ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with edusim gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 74 mg (g TPUK).

MS-ESI: [M+H]+=537,2.

HPLC: Rt=12,09 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 14

tert-Butyl 5-amino-2-(2-thienyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) 5-Cyano-4-(3-nitrophenyl)-2-(2-thienyl)-6-hydroxypyrimidine

A mixture of 2-amidinohydrolase (10.0 g), 3-nitrobenzaldehyde (9.7 g), ethylcyanoacrylate (for 6.81 ml) and potassium carbonate (10.1 g) in absolute EtOH (200 ml) was stirred at 60°C for 8 hours. The reaction mixture was cooled to 0°in the bath with ice, filtered, washed with absolute EtOH and the residue was dissolved in water (100°). The solution was cooled to 50°, acidified 2 N. aqueous solution of HCl to pH 2 and was cooled to 0°in the bath with ice. The precipitate was filtered and washed with ice water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 10,0,

MS-ESI: [M+H]+=325,0.

TLC: Rf=0,3, silica gel, DCM/MeOH=9/1 (V/V).

(b) 6-Chloro-5-cyano-4-(3-nitrophenyl)-2-(2-thienyl)pyrimidine

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-(2-thienyl)-6-hydrox the pyrimidine (example 14(a), 10.0 g) and dimethylaniline (a few drops) in dry 1,4-dioxane (150 ml) was added POCl3(30 ml). After 3 hours at 90°the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane and was carefully added to ice water. The precipitate was filtered and washed with water. The remaining water was removed by joint evaporation with 1,4-dioxane and dried in vacuum at 40°C.

Output: 9,8,

MS-ESI: [M+H]+=343,4.

TLC: Rf=0.8, silica gel, heptane/EtOAc=3/2 (V/V).

(C) Ethyl 5-cyano-4-(3-nitrophenyl)-2-(2-thienyl)-6-(ethoxycarbonylmethyl)pyrimidine

To a stirred solution of ethyl 2-mercaptoacetate (3.28 ml) and 6-chloro-5-cyano-4-(3-nitrophenyl)-2-(2-thienyl)pyrimidine (example 14(b)of 9.8 g) in a mixture of EtOH (80 ml) and DCM (80 ml) under nitrogen atmosphere was added DIPEA (5,57 ml). After 2 hours at room temperature the mixture was diluted with DCM until dissolved, washed with 0.5 N. aqueous HCl solution, dried (MgSO4) and concentrated under reduced pressure.

Output: 12,9,

MS-ESI: [M+H]+=427,2.

TLC: Rf=0,7, silica gel, heptane/EtOAc=3/2 (V/V).

(d) Ethyl 5-amino-4-(3-nitrophenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a stirred solution of ethyl 5-cyano-4-(3-nitrophenyl)-2-(2-thienyl)-6-(ethoxycarbonylmethyl)pyrimidine (example 14(C)of 12.9 g) in a mixture of absolute EtOH (75 ml) and toluene R.A. (75 ml) was added DIPEA (13,0 ml). After 48 hours at 100°the mixture was cooled to 0°C. the precipitate was filtered off, washed with cold EtOH and dried in vacuum at 40°C.

Output: 11,0,

MS-ESI: [M+H]+=427,2.

TLC: Rfor =0.6, silica gel, heptane/EtOAc=3/2 (V/V).

(e) Ethyl 5-amino-4-(3-AMINOPHENYL)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a solution of ethyl 5-amino-4-(3-nitrophenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 14(d), 10,86 g) in 1,4-dioxane (150 ml) was added a solution of tin chloride (II) (15 g) in absolute EtOH (150 ml). Was added 37% aqueous HCl solution (4.5 ml) and the mixture is boiled under reflux for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The residue is suspended in EtOAc (400 ml) and THF was added until complete dissolution. To achieve pH 10-11) was added 4 N. aqueous solution of NaOH. The mixture was diluted by addition of a saturated aqueous solution of NaCl. The organic layer was separated, dried (MgSO4) and concentrated under reduced pressure.

Output: 12,0,

MS-ESI: [M+H]+=397,2.

TLC: Rf=0,4, silica gel, heptane/EtOAc=3/2 (V/V).

(f) 5-Amino-4-(3-nitrophenyl)-2-(2-thienyl)thieno[2,3-d] pyrimidine-6-carboxylic acid

To a solution of ethyl 5-amino-4-(3-AMINOPHENYL)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 14(e)of 10.1 g) in a mixture of 1,4-dioxane (150 ml) and water (50 ml) was added g is droxia potassium (13 g). After 16 hours at 90°the mixture was cooled to 0°C. the precipitate was filtered, suspended in water (180 ml) and cooled to 0°C. the Mixture was acidified to pH 3 by adding 2 N. aqueous citric acid solution, and stirred at 0°C for 2 hours. The precipitate was filtered off, washed with water and dried in vacuum at 40°C.

Output: 6,3,

MS-ESI: [M+H]+=369,2.

TLC: Rf=0,2, silica gel, DCM/MeOH=95/5 (V/V).

(g) tert-Butyl 5-amino-4-(3-nitrophenyl)-2-(2-thienyl)thieno[2,3-d] pyrimidine-6-carboxamide

To a mixture of 5-amino-4-(3-nitrophenyl)-2-(2-thienyl)-thieno[2,3-d] pyrimidine-6-carboxylic acid (example 14(f), 6.3 g) in a mixture of DCM (125 ml) and DMF (N,N-dimethylformamide) (25 ml) under nitrogen atmosphere was added DIPEA (7,1 ml), tert-butylamine (4.3 ml) and TBTU (6.6 g). After 3 hours at room temperature the mixture was diluted with DCM and washed with saturated aqueous NaHCO3, 0.1 N. aqueous solution of HCl and saturated aqueous NaCl. The organic layer was dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel, using as eluent heptane/EtOAc=3/7 to 1/1 (V/V).

Output: 6,45 g

MS-ESI: [M+H]+=424,2.

TLC: Rf=0,3, silica gel, heptane/EtOAc=3/2 (V/V).

(h) tert-Butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-am is but-4-(3-nitrophenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 14(g), 5.0 g) and DIPEA (10.5 ml) in DCM (50 ml) was added bromocatechol (2,40 ml). After 3 hours at room temperature the mixture was diluted with DCM, washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel, using as eluent heptane/EtOAc=3/2 (V/V). Received a mixture of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-thienyl)-thieno[2,3-d]pyrimidine-6-carboxamide and tert-butyl 5-amino-4-(3-(2-chloroacetamido)phenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxamide.

Output: 3,0,

MS-ESI: [M+H]+=546,2, [M,+H]+=500,2.

TLC: Rf=0,2, silica gel, toluene/EtOAc=7/1 (on/about).

(i) tert-Butyl 5-amino-2-(2-thienyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)-phenyl)-2-(2-thienyl)-thieno[2,3-d]pyrimidine-6-carboxamide (example 14(h), 100 mg) in DCM (5 ml) was added N-methyl-2-aminoethanol (140 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, 2x10 ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Decree is Noah in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 59 mg (g TPUK).

MS-ESI: [M+H]+=539,2.

HPLC: Rt=br11.01 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 15

tert-Butyl 5-amino-2-(2-thienyl)-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 14(h), 100 mg) and DIPEA (0.5 ml) in DCM (5 ml) were added hydrochloride methyl ester of glycine (250 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, 2x10 ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 74 mg (g TPUK)

MS-ESI: [M+H]+=553,0

HPLC: Rt=12,57 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 16

tert-Butyl 5-amino-2-(2-thienyl)-4-(3-((N,N-di-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(2-thienyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 14(h), 100 mg) in DCM (5 ml) was added N,N-d-(2-methoxyethyl)-amine (200 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (50 ml), washed with saturated aqueous NaHCO3(1 M, 2x10 ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 59 mg (g TPUK).

MS-ESI: [M+H]+=597,4.

HPLC: Rt=at 13.84 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 17

tert-Butyl 5-amino-2-ethylamino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) tert-Butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-the Mino-2-methylthio-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 4, 1.0 g) in triperoxonane acid (TFUC, 25 ml) was added 3-chloroperbenzoic acid (m-CPBA, 1.0 g). After 17 hours the reaction mixture was concentrated under reduced pressure at ambient temperature (20° (C), re-dissolved in DCM (100 ml), carefully washed with saturated aqueous NaHCO3(G ml) and water (50 ml), dried (MgSO4) and concentrated in vacuum. The crude residue was used in the next stage without additional purification.

Yield: 820 mg

MS-ESI: [M+H]+=547,3.

TLC: Rf=0,2, silica gel, DCM/MeOH=9/1 (V/V).

(b) tert-Butyl 5-amino-2-ethylamino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 17(a), 100 mg) and DIPEA (0.5 ml) in 1,4-dioxane (5 ml) was added ethylamine hydrochloride (150 mg) and the reaction mixture was heated to 60°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% water is th ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 36 mg (g TPUK).

MS-ESI: [M+H]+=528,4.

HPLC: Rt=of 10.21 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 18

tert-Butyl 5-amino-2-(N,N-dimethylamino)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) tert-Butyl 5-amino-2-methanesulfonyl-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methylthio-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 1(i), 1.0 g) in triperoxonane acid (TFUC, 25 ml) was added 3-harperresource acid (m-CPBA, 1.0 g). After 17 hours the reaction mixture was concentrated under reduced pressure at ambient temperature (20° (C), re-dissolved in DCM (100 ml), carefully washed with saturated aqueous NaHCO3(G ml) and water (50 ml), dried (MgSO4) and concentrated in vacuum. The crude residue was used in the next stage without additional purification.

Output: 910 mg.

MS-ESI: [M+H]+=519,6.

TLC: Rf=0,15, silica gel, DCM/MeOH=9/1 (V/V).

b) tert-Butyl 5-amino-2-(N,N-dimethylamino)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 18(a), 100 mg) and DIPEA (0.5 ml) in 1,4-dioxane (5 ml) was added dimethylamine hydrochloride (150 mg) and the reaction mixture was heated to 60°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 36 mg (g TPUK).

MS-ESI: [M+H]+=500,2.

HPLC: Rt=there is a 10.03 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 19

tert-Butyl 5-amino-2-ethylamino-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) tert-Butyl 5-amino-2-methanesulfonyl-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To peremeshivaemogo to a solution of tert-butyl 5-amino-2-methylthio-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 3, 1.0 g) in triperoxonane acid (TFUC, 25 ml) was added 3-chloroperbenzoic acid (m-CPBA, 1.0 g). After 17 hours the reaction mixture was concentrated under reduced pressure at ambient temperature (20° (C), re-dissolved in DCM (100 ml), carefully washed with saturated aqueous NaHCO3(G ml) and water (50 ml), dried (MgSO4) and concentrated in vacuum. The crude residue was used in the next stage without additional purification.

Output: 680 mg.

MS-ESI: [M+H]+=533,6.

TLC: Rf=0,17, silica gel, DCM/MeOH=9/1 (V/V).

(b) tert-Butyl 5-amino-2-ethylamino-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 19(a), 100 mg) and DIPEA (0.5 ml) in 1,4-dioxane (5 ml) was added ethylamine hydrochloride (150 mg) and the reaction mixture was heated to 60°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 1% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 57 mg (g TPUK).

MS-ESI: [M+H]+=514,2.

HPLC: Rt=12,56 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 20

tert-Butyl 5-amino-2-isopropylamino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 17(a), 100 mg) and DIPEA (0.5 ml) in 1,4-dioxane (5 ml) was added Isopropylamine (150 mg) and the reaction mixture was heated to 60°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 57 mg (g TPUK).

MS-ESI: [M+H]+=542,4

HPLC: Rt=br11.01 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 21

tert-Butyl 5-amino-2-allylamino-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 19(a), 100 mg) and DIPEA (0.5 ml) in 1,4-dioxane (5 ml) was added allylamine (200 mg) and the reaction mixture was heated to 60°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 65 mg (g TPUK).

MS-ESI: [M+H]+=to 526.4.

HPLC: Rt=of 13.18 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/waters, the/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 22

tert-Butyl 5-amino-2-methoxy-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 17(a), 200 mg) in methanol (5 ml) was added tert-piperonyl potassium (100 mg) and the reaction mixture was heated to 50°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with an aqueous solution of ammonium chloride (1M, 25 ml), saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 92 mg (g TPUK).

MS-ESI: [M+H]+=515,4.

HPLC: Rt=12,21 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 23

tert-Butyl 5-amino-2-allyloxy-4-(3-((N-ethyl-N-(2-methoxyethyl)-glycinyl)-amino)-phenyl)-ment is about[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)-glycinyl)-amino)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide (example 17(a), 200 mg) in allyl alcohol (5 ml) was added tert-piperonyl potassium (100 mg) and the reaction mixture was heated to 50°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with an aqueous solution of ammonium chloride (1M, 25 ml), saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 63 mg (g TPUK).

MS-ESI: [M+H]+=541,4.

HPLC: Rt=12,71 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 24

tert-Butyl 5-amino-2-isopropoxy-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-2-methanesulfonyl-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)am the but)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 17(a), 200 mg) in isopropanol (5 ml) was added tert-piperonyl potassium (100 mg) and the reaction mixture was heated to 50°C for 3 hours. After concentrating the reaction mixture under reduced pressure, the residue was transferred into DCM (50 ml) and washed with an aqueous solution of ammonium chloride (1M, 25 ml), saturated salt solution (1 M, 25 ml) and water (25 ml). Then the organic layer was dried (MgSO4) and concentrated in vacuum. Thus obtained residue was purified using HPLC, using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 32 mg (g TPUK).

MS-ESI: [M+H]+=543,4.

HPLC: Rt=12,93 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 25

tert-Butyl 5-amino-2-(4-pyridyl)-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

(a) 5-Cyano-4-(3-nitrophenyl)-2-(4-pyridyl)-6-hydroxypyrimidine

A mixture of the hydrochloride of 4-amidinopropane (16.5 g), 3-nitrobenzaldehyde (15.1 g), ethylcyanoacrylate (11.2 ml) and potassium carbonate (16.6 g) in absolute EtOH (250 ml) was stirred at 60°C for 16 hours. The reaction is mesh was cooled to 0° With in the bath with ice. The precipitate was filtered off, washed with absolute EtOH and heated in water (100° (C) to obtain a clear solution. The solution was cooled to 50°C, acidified to pH 2 by adding 2 N. aqueous solution of HCl, and cooled to 0°in the bath with ice. The precipitate was filtered and washed with ice water. The remaining water was removed by joint evaporation with 1,4-dioxane.

Output: 18,3,

MS-ESI: [M+H]+=320,2.

TLC: Rf=0,2, silica gel, DCM/MeOH=9/1 (V/V).

(b) 6-Chloro-5-cyano-4-(3-nitrophenyl)-2-(4-pyridyl)pyrimidine

To a stirred solution of 5-cyano-4-(3-nitrophenyl)-2-(4-pyridyl)-6-hydroxypyrimidine (example 25(a)and 18.3 g) and dimethylaniline (0.5 ml) in dry 1,4-dioxane R.A. (200 ml) was added POCl3(50 ml). After 3 hours at 90°the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane was added ice-cold water. The precipitate was filtered and washed with water. The remaining water was removed by joint evaporation with 1,4-dioxane. Output: 17,2,

MS-ESI: [M+H]+=338,4.

TLC: Rf=0,7, silica gel, heptane/EtOAc=3/2 (V/V).

(C) Ethyl 5-cyano-4-(3-nitrophenyl)-2-(4-pyridyl)-6-(ethoxycarbonylmethyl)pyrimidine

To a stirred solution of 6-chloro-5-cyano-4-(3-nitrophenyl)-2-(4-pyridyl)pyrimidine (example 25(b), and 17.2 g) in a mixture of EtOH (125 ml) and DCM (125 ml) under nitrogen atmosphere EXT is ulali DIPEA (9.8 ml). After 2 hours at room temperature the mixture was diluted with DCM until dissolved, washed with 0.5 N. aqueous HCl solution, dried (MgSO4) and concentrated under reduced pressure.

Output: 20,5,

MS-ESI: [M+H]+=422,0.

TLC: Rfor =0.6, silica gel, heptane/EtOAc=3/2 (V/V).

(d) Ethyl 5-amino-4-(3-nitrophenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a stirred solution of ethyl 5-cyano-4-(3-nitrophenyl)-2-(4-pyridyl)-6-(ethoxycarbonylmethyl)pyrimidine (example 25(C)of 20.5 g) in a mixture of absolute EtOH (100 ml) and toluene R.A. (100 ml) was added DIPEA (20,0 ml). After 48 hours at 100°the mixture was cooled to 0°C. the precipitate was filtered off, washed with cold EtOH and dried in vacuum at 40°C.

Output: 15,7,

MS-ESI: [M+H]+=422,2.

TLC: Rf=0,5, silica gel, heptane/EtOAc=3/2 (V/V).

(e) Ethyl 5-amino-4-(3-AMINOPHENYL)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxylate

To a solution of ethyl 5-amino-4-(3-nitrophenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 25(d), 15.7 g) in 1,4-dioxane R.A. (250 ml) was added a solution of tin chloride (II) (21,0 g) in absolute EtOH (250 ml). Was added 37% aqueous HCl solution (6.9 ml) and the mixture was heated under reflux (90° (C) for 16 hours. The mixture was left to cool to room temperature and concentrated under reduced pressure. The residue is suspended in EtOAc (500 ml). To achieve ur is una pH 10-11) was added 4 N. an aqueous solution of NaOH. The mixture was diluted by addition of a saturated aqueous solution of NaCl. The organic layer was separated, dried (MgSO4) and concentrated under reduced pressure.

Output: 12,0,

MS-ESI: [M+H]+=392,2.

TLC: Rf=0,3, silica gel, heptane/EtOAc=3/2 (V/V).

(f) 5-Amino-4-(3-nitrophenyl)-2-(4-pyridyl)thieno[2,3-d] pyrimidine-6-carboxylic acid

To a solution of ethyl 5-amino-4-(3-AMINOPHENYL)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxylate (example 25(e), 12.0 g) in a mixture of 1,4-dioxane (210 ml) and water (80 ml) was added potassium hydroxide (15.7 g). After 16 hours at 90°the mixture was cooled to 0°C. the precipitate was filtered, suspended in water (300 ml) and cooled to 0°C. the Mixture was acidified to pH 3 by adding 2 N. aqueous citric acid solution, and stirred at 0°C to room temperature for 2 hours. The precipitate was filtered off, washed with water and dried in vacuum at 40°C.

Output: 12,0,

MS-ESI: [M+H]+=364,2.

TLC: Rf=0,1, silica gel, DCM/MeOH=95/5 (V/V).

(g) tert-Butyl 5-amino-4-(3-AMINOPHENYL)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a mixture of 5-amino-4-(3-AMINOPHENYL)-2-(4-pyridyl)thieno[2,3-d] pyrimidine-6-carboxylic acid (example 25(f), 12.0 g) in a mixture of DCM (250 ml) and DMF (50 ml) under nitrogen atmosphere was added DIPEA (12.9 ml), tert-butylamine (7.8 ml) and TBTU (11.9 g). After 2 hours at room temperature the mixture rasba the Lyali DCM and washed with saturated aqueous NaHCO 3, 0.1 N. aqueous solution of HCl and saturated aqueous NaCl. The organic layer was dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by chromatography on silica gel, using as eluent dichloromethane/methanol=1/0 to 95/5 (V/V).

Output: 12,2,

MS-ESI: [M+H]+=419,4.

TLC: Rf=0,3, silica gel, heptane/EtOAc=3/2 (V/V).

(h) tert-Butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a solution of tert-butyl 5-amino-4-(3-nitrophenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 25(g), 2.0 g) and N,N-dimethylaniline (3.0 ml) in THF (50 ml) was added dropwise a solution of bromoacetamide (0,59 ml) in THF (25 ml). After 30 min at room temperature the mixture was concentrated under reduced pressure, then dissolved in DCM (100 ml), washed with saturated aqueous NaHCO3, dried (MgSO4) and concentrated in vacuum. The residue was used in the next stage without additional purification.

Output: 2,3,

MS-ESI: [M+H]+=539,2.

TLC: Rf=0,3, silica gel, heptane/EtOAc=3/2 (V/V).

(i) tert-Butyl 5-amino-2-(4-pyridyl)-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)-phenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 25(h), 200 mg) is DCM (5 ml) was added 2-amino-2-methylpropanol (300 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (100 ml), washed with saturated aqueous NaHCO3(1 M, h ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% aqueous TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 73 mg (g TPUK).

MS-ESI: [M+H]+=548,2.

HPLC: Rt=9,65 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 26

tert-Butyl 5-amino-2-(4-pyridyl)-4-(3-((N,N-bis-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide

To a stirred solution of tert-butyl 5-amino-4-(3-(2-bromoacetamide)phenyl)-2-(4-pyridyl)thieno[2,3-d]pyrimidine-6-carboxamide (example 25(h), 200 mg) in DCM (5 ml) was added bis-(2-methoxyethyl)amine (300 mg). After stirring for 17 hours at room temperature the reaction mixture was diluted with DCM (100 ml), washed with saturated aqueous NaHCO3(1 M, h ml), dried (MgSO4) and concentrated in vacuum. The residue was purified via HPLC using a column Luna C-18 with the following gradient: 0.1% the command TFUK + 10% aqueous ACN/ACN=90/10 to 10/90 (V/V) for 30 minutes Specified in the title compound was then liofilizovane of a mixture of 1,4-dioxane, 0.1% aqueous TFUK and water.

Yield: 170 mg (g TPUK).

MS-ESI: [M+H]+=592,2.

HPLC: Rt=12,02 min, column Luna C-18(2), 3 μm, h,0 mm, UV detection=210 nm, oven temperature=40°C, flow=0.25 ml/min, eluent: phosphate buffer 50 mm pH 2,1/water/ACN=10/80/10 to 10/10/80 (about/about/about), cycle time=20 minutes

Example 27

CHO-LH and CHO-FSH bioactivityin vitro

The activity of compounds as agonists LH felt ovary cells Chinese hamster (Chinese Hamster Ovary (CHO)), stable tropicabana LH receptor of human and karanfilovski element (CRE), which is sensitive to cAMP/promoter directing the expression of the reporter gene Firefly luciferase. The binding of ligand to Gs-coupled receptor LH results in an increase of camp, which, in turn, induces an increased transactivation luciferase reporter construct. Quantification of luciferase signal was performed using a luminescence counter. For test compounds were calculated value EU50(concentration of test compound that causes premaxilla (50%) stimulation). For this purpose used the program GraphPad PRISM version 3.0 (GraphPad software Inc., San Diego).

Similarly the activity of compounds as agonists of FSH tested the cells and Cho, tropicabana reporter gene luciferase and FSH receptor of human rights. The results are presented in table 1.

The bioactivityin vivo

To measurein vivothe activity of compounds which are agonists of the receptor, LH/FSH, investigated the induction of ovulation in immature mice. In this analysis of immature female mice were primiraly isolated from urine FSH (Humegon of 12.5 IU/animal). After approximately 48 hours the animals were treated with compounds agonists LH/FSH, which are described in examples 1, 4, 9 and 17, with a dose of 50 mg/kg Animals were killed 24 hours after treatment LH/FSH agonist and under a microscope and counted the number of germ cells in the fallopian tube. On average was investigated 10-15 animals. The average number counted germ cells was 8, except for the compounds of example 17 (where this number amounted to 0.4).

Table 1
NameIf-

measures
LH EC50(M)FSH EC50(M)
tert-Butyl 5-amino-2-methylthio-4-

(3-((N-methyl-N-(2-hydroxyethyl)-

glycidyl)amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
12,E-095,E-08
tert-Butyl 5-amino-2-methylthio-4-

(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno,3-d]pyrimidine-6-carboxamide
25,E-098,06A-07
tert-Butyl 5-amino-2-methylthio-4-(3-((N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
35,E-091,44TH-06
tert-Butyl 5-amino-2-methylthio-

4-(3-((N-ethyl-N-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
47,E-091,E-07
tert-Butyl 5-amino-2-methylthio-4-

(3-((N-(R-1-methoxycarbonyl-2-

methylprop-1-yl)glycinyl)amino)-

phenyl)thieno[2,3-d]pyrimidine-6-

carboxamid
59,E-093,48TH-07
tert-Butyl 5-amino-2-methylthio-4-

(3-((N,N-bis-(2-methoxyethyl)glycinyl)-

amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
61,E-081,18E-06
tert-Butyl 5-amino-2-methylthio-4-

(3-((2,3-dihydroxypropyl-1-yl)glycinyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
71,22E-081,the 13TH-06
tert-Butyl 5-amino-2-methylthio-4-

(3-((1,3-dihydroxypropyl-2-yl)glycinyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
82,E-081,E-06
tert-Butyl 5-amino-2-phenyl-4-

(3-((N-ethyl-N-(2-g is proxetil)-

glycidyl)amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
94,E-096,06A-07
tert-Butyl 5-amino-2-phenyl-4-

(3-(N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
101,E-082,48TH-06
tert-Butyl 5-amino-2-(2-furyl)

-4-(3-((N-methyl-N-(2-hydroxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
114,E-096,E-07
tert-Butyl 5-amino-2-(2-furyl)-

4-(3-(N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
121,18E-082,E-06
tert-Butyl 5-amino-2-(2-furyl)-

4-(3-(N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
131,E-082,E-06
tert-Butyl 5-amino-2-(2-thienyl)-

4-(3-((N-methyl-N-(2-hydroxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
145,28TH-091,44TH-06
tert-Butyl 5-amino-2-(2-thienyl)-

4-(3-((N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
151,E-08tert-Butyl 5-amino-2-(2-thienyl)-

4-(3-((N,N-di-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
162,the 13TH-086,E-06
tert-Butyl 5-amino-2-ethylamino-

4-(3-((N-ethyl-N-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
171,E-081,25TH-06
tert-Butyl 5-amino-2-(N,N-

dimethylamino)-4-(3-((N-methyl-

N-(2-hydroxyethyl)glycinyl)-

amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
181,E-081,E-06
tert-Butyl 5-amino-2-ethylamino

-4-(3-((N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
192,E-082,30E-06
tert-Butyl 5-amino-2-isopropylamino-4-(3-((N-ethyl-

N-(2-methoxyethyl)glycinyl)-

amino)phenyl)thieno[2,3-d]-

pyrimidine-6-carboxamide
204,24TH-082,E-06
tert-Butyl 5-amino-2-allylamino-

4-(3-((N-(methoxycarbonylmethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
215,the 70S-085,the ' 60S-07
tert-Butyl 5-amino-2-methoxy-

4-(3-((N-ethyl-N-(2-methoxyethyl)-
glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
221,E-089,A-07
tert-Butyl 5-amino-2-allyloxy-

4-(3-((N-ethyl-N-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
233,E-081,74A-06
tert-Butyl 5-amino-2-isopropoxy-

-4-(3-((N-ethyl-N-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
247,E-082,E-06
tert-Butyl 5-amino-2-(4-pyridyl)-

-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)-

thieno[2,3-d]pyrimidine-6-carboxamide
253,E-084,E-07
tert-Butyl 5-amino-2-(4-pyridyl)-

-4-(3-((N,N-bis-(2-methoxyethyl)-

glycidyl)amino)phenyl)thieno-

[2,3-d]pyrimidine-6-carboxamide
263,E-081,23RD-06

1. Derived thieno[2,3-d]pyrimidine of the General formula I

or its pharmaceutically acceptable salt,

where X represents O or H,H;

And represents S, NH, N(R6), Or the connection;

R1represents (1-4C)alkyl, (2-4C)alkenyl, unsubstituted phenyl or unsubstituted furyl, thienyl, pyridyl;

R3and R4independently selected from H, (1-4C)alkyl and hydroxy(1-4C)alkyl;

R5represents H or (1-4C)alkyl and

R6represents (1-4C)alkyl.

2. The compound according to claim 1, in which X represents N,N.

3. The compound according to claim 1 or 2, in which R5represents (1-4C)alkyl.

4. The compound according to claim 1 or 2, in which R3=R4.

5. The compound according to claim 1 or 2, in which R3=R4=H.

6. The compound according to claim 1 or 2, in which R2represents (1-4C)alkyl.

7. The compound according to claim 1 for use in therapy as a means to control fertilization.

8. Pharmaceutical composition, controlling fertilization, including thieno[2,3-d]pyrimidine according to any one of claims 1 to 6, or its pharmaceutically acceptable salt, or MES in a mixture with a pharmaceutically acceptable auxiliary substance.

9. The use of thieno[2,3-d]pyrimidine according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, or MES for production of medicines for the control of fertilization.

10. A method of treating disorders associated with fertilization, patients in need, including the introduction of an effective amount of a compound according to any of claims 1 to 6.

11. The connection selected is from the group including

tert-butyl 5-amino-2-methylthio-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl}thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((N-(R-1-methoxycarbonyl-2-methylprop-1-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((N,N-bis-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((2,3-dihydroxypropyl-1-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methylthio-4-(3-((1,3-dihydroxypropyl-2-yl)glycinyl)amino)phenyl-thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-phenyl-4-(3-((N-ethyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-phenyl-4-(3-(N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-furyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-furyl)-4-(3-(N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-furyl)-4-(3-(N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-thienyl)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-thienyl)-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(2-thienyl)-4-(3-((N,N-di-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-ethylamino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(N,N-dimethylamino)-4-(3-((N-methyl-N-(2-hydroxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-ethylamino-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-isopropylamino-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-allylamino-4-(3-((N-(methoxycarbonylmethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-methoxy-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-allyloxy-4-(3-((N-ethyl-N-(2-methoxyethyl)-glycidyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-isopropoxy-4-(3-((N-ethyl-N-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(4-pyridyl)-4-(3-((N-(1-hydroxy-2-methylprop-2-yl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide;

tert-butyl 5-amino-2-(4-pyridyl)-4-(3-((N,N-bis-(2-methoxyethyl)glycinyl)amino)phenyl)thieno[2,3-d]pyrimidine-6-carboxamide,

or its pharmaceutically acceptable salt.



 

Same patents:

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

SUBSTANCE: invention describes compounds of the formula (I):

wherein R1 means hydrogen atom; R2 means phenyl or phenyl mono- or di-substituted substituted with the following group: halogen atom, (lower)-alkyl, (lower)-alkoxy-group, perfluoro-(lower)-alkyl; R3 and R4 in common with carbon atoms to which they are bound form phenyl optionally and independently mono-, di- or tri-substituted with halogen atom or perfluoro-(lower)-alkyl, or form 5-, 6- or 7-membered saturated cycle optionally comprising heteroatom chosen from oxygen (O) and sulfur (S) atom and optionally and independently mono-substituted with (lower)-alkyl wherein indicated saturated cycle is condensed in ortho-position with 5-membered aromatic cycle optionally comprising S atom as a heteroatom, or with phenyl optionally and independently mono- di-substituted with the group: halogen atom, (lower)-alkyl, perfluoro-(lower)-alkyl or (lower)-alkoxy-group, and their pharmaceutically acceptable salts. Also, invention describes a method for synthesis of compounds, a pharmaceutical composition and using compounds for treatment and/or prophylaxis of DPP-IV-associated diseases. Compounds are used in treatment of such diseases as diabetes mellitus being first of all non-insulin dependent diabetes mellitus and damaged tolerance to glucose.

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

16 cl, 1 tbl, 39 ex

FIELD: agriculture, organic chemistry.

SUBSTANCE: invention relates to application of 2-[N-(2'-iodophenyl)carboxamido]-3-amino-4,6-dimethylthieno[2,3-b]pyridine of formula as stimulator of sunflower growth.

EFFECT: stimulation of sunflower seed germination; increased sunflower productivity.

2 tbl, 3 ex

FIELD: organic chemistry of heterocyclic compounds, pharmacy.

SUBSTANCE: invention relates to new bicyclic heteroaromatic compounds of the general formula (I): wherein R1 represents phenyl optionally substituted with NHR5 or OR5; R2 represents (C1-C4)-alkyl or phenyl; R5 represents phenylcarbonyl, (C4-C6)-heterocycloalkylcarbonyl, (C2-C8)-alkenylsulfonyl and others; Y represents nitrogen atom (N); Z represents -NH2 or -OH. A represents sulfur atom (S) or a bond; B represents -N(H) or oxygen atom (O); X1-X2 represent C=C, -NH-C(O), C=N and others; Proposed compounds show agonistic activity with respect to LH receptor and can be used in medicine.

EFFECT: valuable medicinal properties of compounds.

10 cl, 34 ex

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to new derivatives of β-carboline of the general formula (I)

showing properties of phosphodiesterase V inhibitor (PDE V). In the general formula (I) R1 means hydrogen atom; n = 0; X is taken among the group consisting of oxygen (O), sulfur (S) atoms and NRD; R2 is taken among the following group: phenyl (that can be optionally substituted with 1-3 RB), 6-membered nitrogen-containing heteroaryl and 5-6-membered heterocycloalkyl comprising 1-2 oxygen atoms and condensed with benzene ring (optionally substituted with 1-3 RB); R4 is taken among the group consisting of hydrogen atom, carboxy-group. (C1-C6)-alkylcarbonyl, di-[C1-C8)-alkyl]-aminoalkoxycarbonyl, di-[(C1-C8)-alkyl]-amino-(C1-C8)-alkylaminocarbonyl; a = a whole number from 0 to 1; Y is taken among the group consisting of -CH2, -C(O); Z is taken among the group consisting of -CH2, -CHOH, and -C(O) under condition that when Z represents -CHOH or -C(O) then X represents -NH; is taken among the group consisting of naphthyl, 5-6-membered heteroaryl comprising 1-3 heteroatoms taken among nitrogen, oxygen and/or sulfur atoms possibly condensed with benzene ring; m = a whole number from 0 to 2; R3 is taken independently among the group consisting of halogen atom, nitro-group, (C1-C8)-alkyl, (C1-C8)-alkoxy-group, trifluorophenyl, phenyl (optionally substituted with 1-3 RB), phenylsulfonyl, naphthyl, (C1-C8)-aralkyl, 5-6-membered heteroaryl comprising 1-3 nitrogen atoms in the ring (optionally substituted with 1-3 RB). Also, invention relates to a pharmaceutical composition, a method for its preparing and methods for inhibition of phosphodiesterase V activity (PDE V), and for increase of the cGMP concentration.

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

14 cl, 11 sch, 7 tbl, 13 ex

FIELD: biochemistry, medicine, in particular new bioactive compounds having peptide hormone vasopressin agonistic activity.

SUBSTANCE: disclosed are compounds of general formula 1 or 2 or tautomers, or pharmaceutically acceptable salts thereof, wherein W represents N or C-R4; R1-R4 are independently H, F, Cl, Br, alkyl, O-alkyl, NH2, NH-alkyl, N(alkyl)2, NO2 or R2 and R3 together may form -CH=CH-CH=CH-; G1 represents bicyclic or tricyclic condensed azepine derivatives selected from general formulae 3-8 wherein A1, A4, A7, and A10 are independently CH3, O, and NR5; A2, A3, A9, A11, A12, A14, and A15 are independently CH and N; or A5 represents covalent bond and A6 represents S; or A5 represents N=CN and A6 represents covalent bond; A8 and A12 are independently NH, N-CH3 and S; A16 and A17 both represent CH2 or one of A16 and A17 represents CH2 and the other represents CH(OH), CF2, O, SOa, and NR5; R5 represents H, alkyl, CO-alkyl, and (CH2)bR6; R6 represents phenyl, pyridyl, OH, CO2H; a = 0-2; b = 1-4; Y represents CH or N; Z represents CH=CH or S; and G2 represents group selected from groups of formulae 9-11 wherein Ar represents phenyl, pyridyl, naphthyl, and mono- or polysubstituted phenyl, pyridyl, wherein substituents are selected from F, Cl, Br, alkyl, NO2; D represents covalent bond or NH; E1 and E2 both are H, OMe, F, or one of E1 and E2 represents OH, O-alkyl, OBn, OPh, OAc, F, Cl, Br, N2, NH2, NHBn or NHAc and the other represents H; or E1 and E2 together form =O, -O(CH2)gO- or -S(CN2)gS-; F1 and F2 both represent H or together form =O or =R; L represents OH, O-alkyl, NH2, NH-alkyl, and NR9R10; R7 represents COR8; R8 represents OH, O-alkyl, NH2, NH-alkyl, N(alkyl)2, pyrolidinyl, and piperidinyl; R9 and R10 both are alkyl or together form -(CH2)h-; V represents O, N-CN or S; c = 0 or 1; d = 0 or 1, e = 0 or 1; f = 0-4; g = 2 or 3; h = 3-5, with the proviso, that both d and e are not 0. Also disclosed are pharmaceutical composition having agonistic activity in relate to V2 receptor, method for treatment one or more diseases (e.g., enuresis, nycturia, diabetes insipidus, hemorrhage disorders, urinary incontinence.

EFFECT: new compounds with value biological characteristics.

41 cl, 19 tbl, 193 ex

FIELD: organic chemistry, medicine, hematology.

SUBSTANCE: invention elates to new compounds that inhibit activated blood coagulating factor X (Fxa factor) eliciting the strong anti-coagulating effect. Invention proposes compound of the formula (1): Q1-Q2-C(=C)-N-(R1)-Q3-N(R2)-T1-Q4(1) wherein R1, R2, Q1, Q2, Q4 and T1 have corresponding values, and Q2 represents the group of the formula: wherein R9, R10 and Q5 have corresponding values also, or its salt, solvate or N-oxide. Invention provides the development of a novel compound possessing strong Fxa-inhibiting effect and showing the rapid, significant and stable anti-thrombosis effectin oral administration.

EFFECT: valuable medicinal properties of compounds.

13 cl, 1 tbl, 195 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new nitrogen-containing aromatic derivatives of the general formula:

wherein Ag represents (1) group of the formula:

; (2) group represented by the formula:

or ; (3) group represented by the formula:

; Xg represents -O-, -S-, C1-6-alkylene group or -N(Rg3)- (wherein Rg3 represents hydrogen atom); Yg represents optionally substituted C6-14-aryl group, optionally substituted 5-14-membered heterocyclic group including at least one heteroatom, such as nitrogen atom or sulfur atom, optionally substituted C1-8-alkyl group; Tg1 means (1) group represented by the following general formula:

; (2) group represented by the following general formula: . Other radical values are given in cl. 1 of the invention claim. Also, invention relates to a medicinal agent, pharmaceutical composition, angiogenesis inhibitor, method for treatment based on these compounds and to using these compounds. Invention provides preparing new compounds and medicinal agents based on thereof in aims for prophylaxis or treatment of diseases wherein inhibition of angiogenesis is effective.

EFFECT: improved treatment method, valuable medicinal properties of compounds and agents.

40 cl, 51 tbl, 741 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel biologically active compounds. Invention describes compounds or their salts of the general formula (I): A-B-N(O)s (I) wherein s = 2; A means R-T1- wherein R represents radical of a medicinal substance under condition that a medicinal substance by the formula R-T1-Z or R-T1-OZ wherein Z represents hydrogen atom (H) or (C1-C5)-alkyl is taken among paracetamol, salbutamol, ambroxol, alendronic acid,, cetirizine, ampicillin, aciclovir, doxorubicin, simvastatin, diphylline, tacrine, clopidogrel, dimethylomeprazol, diclofenac, ferulic acid, enalapril, propranolol, benfurodil hemisuccinate, tolrestate or sulindac; T1 means (CO), oxygen atom (O) or NH; B means TB-X2-O- wherein TB means bivalent radical R1B-X-R2B wherein R1B and R2B are similar or different and represent linear or branched (C1-C6)-alkylenes and X represents a bond, oxygen (O), sulfur (S) atom or NR1C wherein NR1C represents hydrogen atom (H) or linear or branched (C1-C6)-alkyl; corresponding precursor B is represented by the formula -TB-X2-OH wherein TB means (CO) and free valence in TB represents -OZ wherein Z is determined above, or TB means oxygen atom (O), and free valence in TB represents hydrogen atom (H) under condition that in the formula (I) when X2 in precursor B represents linear or branched (C2-C20)-alkylene then a medicinal substance by the formula R-T1-Z or R-T1-OZ used in the formula (I) doesn't belong to the following substances: enalapril (ACE inhibitors) and diclofenac (NSAID). Also, invention describes pharmaceutical compositions for using in cases of oxidative stress and 4-nitroxybutanoic acid 4'-acetylaminophenyl ester. Invention provides preparing novel compounds possessing useful biological properties.

EFFECT: valuable medicinal properties of medicinal substances and compositions.

7 cl, 8 tbl, 32 ex

FIELD: organic chemistry, medicine, biochemistry, pharmacy.

SUBSTANCE: invention relates to 2-aminomethylthieno[2,3-d]pyrimidines of the general formula (I): wherein R1 and R2 in common with C-atoms with which they are bound form 5-7-membered monounsubstituted cycloalkenyl ring; R3 and R4 are similar or different and represent independently of one another (C1-C8)-alkoxy-group or halogen atom; R5 and R6 can be similar or different and represent independently of one another hydrogen atom, linear or branched (C1-C8)-alkyl group that can be substituted with one or more hydroxyl, (C1-C8)-alkoxy-group, amine, mono-(C1-C8-alkyl)-amine or di-(C1-C8-alkyl)-amine groups, or in common with nitrogen atom to which they are bound form a heterocyclic ring that comprises optionally one or more additional nitrogen atoms and substituted with one or more hydroxyl, (C1-C8)-alkoxy- or (C1-C8)-alkylol groups. Compounds elicit the inhibitory effect with respect to activity of phosphodiesterase V and can be used in treatment of cardiovascular system states and in disturbance in the potency injury. Also, invention describes a medicinal preparation based on compounds said, a method for its preparing and a method for preparing compounds.

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

6 cl, 1 tbl, 16 ex

FIELD: pharmaceutical chemistry, medicine.

SUBSTANCE: invention relates to substituted pyridines and pyridazines with angiogenesis inhibition activity of general formula I

(I)1, wherein ring containing A, B, D, E, and L represents phenyl or nitrogen-containing heterocycle; X and Y are various linkage groups; R1 and R2 are identical or different and represent specific substituents or together form linkage ring; ring J represents aryl, pyridyl or cycloalkyl; and G's represent various specific substituents. Also disclosed are pharmaceutical composition containing claimed compounds, as well as method for treating of mammalian with abnormal angiogenesis or treating of increased penetrability using the same.

EFFECT: new pyridine and pyridazine derivatives with angiogenesis inhibition activity.

26 cl, 6 tbl, 114 ex

FIELD: organic chemistry, steroids, biology.

SUBSTANCE: invention relates to steroid compounds of the general formula (X):

wherein in fragment of the formula XA:

each bond between C6 and C7, between C7 and C8, between C8 and C9, between C8 and C14 and between C14 and C15 is a single or double bond under condition that each atom C6, C7, C8, C9, C14 and C15 is bound with adjacent C-atom by a single bond or one double bond; CR3 means -CHOH; A means methylene or ethylene group; R4 and R4' mean (C1-C4)-alkyl, hydrogen atom (H); R20 means (C1-C4)-alkyl; R23 and R23' mean in common piperidine-1-yl, morpholine-4-yl, pyrrolidine-1-yl, piperazinyl possibly substituted with -OH, benzene, pyridine, pyrimidine, phenyl, alkoxycarbonyl group, or R23 means H and R23' means substituted alkyl. These compounds can be used for stimulation of meiosis in human oocytes. In proposed compounds steroid differs specifically as nitrogen atom of amino-group is bound with C17-atom of steroid skeleton by spacer A.

EFFECT: improved methods of synthesis, valuable biological properties of compounds.

16 cl, 8 dwg, 2 tbl, 30 ex

FIELD: medicine, gynecology.

SUBSTANCE: invention relates to a method for rehabilitation of women after medicinal artificial abortion. Method involves prescription of the preparation "Multi-tabs-intensiv" in the first 24 h after expulsion of fetus in the dose 1 tablet for 1 month and additional administration of the preparation "Djufaston" from 16-th to 25-th days in the dose 10 mg, 2 times per a day. Method provides the full-value recovery of endometrium in I-II phases of cycle after medicinal artificial abortion under conditions of blocking receptors to progesterone at the level of uterus as target-organ and recovery of physiological menstrual cycle.

EFFECT: improved method for rehabilitation.

1 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to anhydrous, antifungal, lubricating gel compositions comprising polyhydric alcohol, gelatinizing agent and an antifungal azole compound, and to a method for treatment of a patient with fungal infections comprising administration of indicated composition in a patient. Compositions shows the excellent warming and lubricating effect after its applying on skin and mucosal tissues and provides the effective treatment of fungal infections.

EFFECT: improved and valuable properties of compositions.

21 cl, 2 tbl, 11 dwg, 9 ex

FIELD: medicine, gynecology.

SUBSTANCE: patients should be suggested to take radon procedures as baths, gynecolopgical irrigations, microenemas, laserotherapy in autoresonance mode at altering frequency of laser apparatus independently against frequency characteristics of human body, at wave length of 0.89 mcm, power of 10 W onto area of uterine projection in case of genital endometriosis and uterine myoma at exposure of 15 min and onto mammary glands in case of diffuse proliferative mastopathies. Impact fields should be chosen by sectors ranged 10-16 per 10-15 sec for each impact field. Local therapy should be conducted about 2.5-3 h after taking radon procedures successively, not earlier. The first stage - baths with malavit at 1:5 dilution at exposure of 15 min. The second stage - after removing the solution out of vagina it is necessary to introduce malavit gel at a tampon for 4-6 h. The whole therapy should be carried out together with internal intake of mineral water 30-60 min before meals thrice daily per 3.5 mg/kg body weight. The innovation enables to achieve prolonged remission, normalize hormonal level in blood and duration of the second phase of the cycle and decrease cholesterol level in blood.

EFFECT: higher efficiency of correction.

1 dwg, 4 ex

FIELD: medicine, gynecology.

SUBSTANCE: as a suggested preparation one should apply dried defibrinated or non-defibrinated blood of Siberian stags or spotted deer. The preparation suggested should be prescribed perorally per 25 mg (1 capsule) twice daily, therapy course corresponds to 25-35 d. Application of this preparation in women at normal level of gonadotropins, positive gestagen-test, with no hyperandrogenia and hyperprolactinemia at the absence of menstrual cycle of more than 3 mo provides the onset of menstruations at stable positive effect. After carrying out 3 therapeutic courses no relapses had been observed.

EFFECT: higher efficiency for regulating menstrual cycle.

3 cl, 2 ex, 3 tbl

FIELD: organic chemistry of heterocyclic compounds, pharmacy.

SUBSTANCE: invention relates to new bicyclic heteroaromatic compounds of the general formula (I): wherein R1 represents phenyl optionally substituted with NHR5 or OR5; R2 represents (C1-C4)-alkyl or phenyl; R5 represents phenylcarbonyl, (C4-C6)-heterocycloalkylcarbonyl, (C2-C8)-alkenylsulfonyl and others; Y represents nitrogen atom (N); Z represents -NH2 or -OH. A represents sulfur atom (S) or a bond; B represents -N(H) or oxygen atom (O); X1-X2 represent C=C, -NH-C(O), C=N and others; Proposed compounds show agonistic activity with respect to LH receptor and can be used in medicine.

EFFECT: valuable medicinal properties of compounds.

10 cl, 34 ex

FIELD: medicine.

SUBSTANCE: the present innovation deals with composition for fertilization in vitro and the system for its delivering (device). The composition suggested contains steroid at the quantity of below 5% (weight/weight), that is: 4.4-dimethyl-5α-cholesta-8.14.24-trien-3β-ol, hemisuccinate of 4.4-dimethyl-5α-cholest-8.14.24-trien-3β-ol; 5α-cholest-8.14-dien-3β-ol; hemisuccinate of 5α-cholest-8.14-dien-3β-ol; (20S)-cholest-5-en-3, 20-diol; N-(methionine)amide of 3β-hydroxy-4.4-dimethyl-5α-chol-8.14-dien-24-oic acid or cholest-5-en-16β-ol, and, also, additive (water-soluble protein or phosphoglyceride). Delivering system has got either one foramen or one cavity that contains the composition mentioned as a solid product or solution. The composition of sterols contains no constituents negatively affecting oocytes and could be dissolved in aqueous medium without physical impact (that is, heating, mixing or ultrasound treatment).

EFFECT: higher efficiency of fertilization in vitro.

8 cl, 5 ex, 3 tbl

FIELD: medicine, urology.

SUBSTANCE: at the background of curative starvation a patient should be prescribed for peroral intake of 100 ml tincture or 1 teaspoon tincture out of bees twice daily in the morning and in the evening, and, also, 200 ml hempseed decoction based upon milk twice daily in the morning and in the evening at the course of 5 d. On the 3d, 5th, 7th d of starvation one should perform hepatic and biliary ductal purification at 7 p.m. at the intake of 250 ml natural apple juice at 4 p.m., since the 1st to the 7th d of starvation one should daily apply enemas: in the morning - purifying enema with juice of 1 lemon, in the daytime - curative enema with Artemisia decoction. One should daily perform rectal prostatic massage followed by applying microenemas with 30 ml composition containing the tincture of Sophora, oil of wheat germs, decoction of medicinal plant of antiphlogistic action taken at weight ratio of 1:0.5:2, correspondingly, at the course of 10 d long. Duration of curative starvation lasts for 7 d, not less. The method provides shortened terms of therapy due to simultaneous normalization of body metabolic processes, purification of organs against bacteria and parasites.

EFFECT: decreased body allergization.

6 cl, 2 ex

FIELD: veterinary science.

SUBSTANCE: on should apply 1-(ethoxy)sylatrane as a stimulating agent for reproductive capacity in female furred animals and viability of their offspring. The innovation enables to improve reproductive function in females and viability of whelps.

EFFECT: higher efficiency.

3 ex, 3 tbl

FIELD: pharmaceutical industry.

SUBSTANCE: invention provides therapeutical formulation for improving fertility and treating sterility caused by polycystic ovary syndrome, which composition is characterized by that formulation thereof involves use of dipeptidylpeptidase IV inhibitor. Treatment of polycystic ovary syndrome-caused sterility is high successful with effective active ingredient concentration equal to 1 μM. When dipeptidylpeptidase IV inhibitor is administered, luteinizing hormone and testosterone levels in blood are normalized. Claimed compounds are low toxic for warm-blooded animals and humans.

EFFECT: widened choice of efficient antisterility drugs.

7 cl, 3 ex

FIELD: medicine, oncology, radiology.

SUBSTANCE: the suggested thermochemoradiation treatment should be fulfilled in three stages at intervals between each of them being 10-14 d: during the 1st stage it is necessary to carry out distance gamma-therapy daily at fractionating the dosage during 1-3 d per 4 Gy, then during 4-22 d per 2 Gy up to TFD being 50 isoGy, radiosensitization with 5-fluorouracil daily at the dosage of 125 mg intravenously at flow-type technique 30 min before the séance of radiation treatment and local thermotherapy with transrectal sensor once/2 d, every other day, just before gamma-therapy; during the 2nd stage it is necessary to conduct radiosensitization and local thermotherapy similarly, as it was during the first stage, as for irradiation it should be carried out as intracavitary gamma-therapy daily per 3 Gy up to TFD being 18-24 Gy; during the 3d stage one should fulfill radiosensitization with fluorouracil as during the first and second stages and carry out distance gamma-therapy daily per 2 Gy so that TFD from all three stages should achieve 74 isoGy.

EFFECT: higher efficiency of therapy.

3 ex

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