Taxanes, method for their production and pharmaceutical drugs based on them

 

(57) Abstract:

Describes the new taxanes General formula I, with pharmacological activity, where Skcan be a , R1can represent hydrogen or C1- C10-acyl, R2may represent-OH, R3can be a C1- C10-alkoxygroup, and the free hydroxy-group in the formula I can be further functionally modified by esterification. Describes the method of obtaining them and pharmaceutical drugs based on them. In addition to the increased stability of microtubules, the compounds of formula I have additional effects on the polymerization of tubulin. Can be used for the treatment of malignant tumors. 3 s and 5 C.p. f-crystals, 4 Il., 1 PL.

The invention relates to pharmacologically active compounds, which have the ability to influence the processes of polymerization and depolymerization of tubulin.

The number of blockers mitosis of natural origin used as anticancer agents, or expose currently in clinical testing. There are different classes of such inhibitors of mitosis, which Tannoy system (for example, the Vinca alkaloids (Vinka alkaloids, colchicine), or exert their cytotoxic effect by the GTP-independent increase the polymerization of tubulin and prevent depolymerization of microtubules (e.g., Taxol, Taxotere (taxoters)).

Due to its physicochemical properties, the remaining misunderstood so far, and as a consequence of the properties of tumor cells, inhibitors of mitosis have a certain selectivity to tumor cells, but they still have a significant cytotoxic effect on normal cells. Search more selectively acting compounds, the production of which would be easier to establish, and which, as the connection class taxan, is able to inhibit the depolymerization of microtubules, unexpectedly led to the discovery of esters of borneol, as described in German patent N P 4416374.6 and C 07 C 35/30, 5.05.94 (Schering AG) and German patent N 19513040.5, C 07 D 303/06, 29.03.95 (Schering AG). Structural modification of compounds of this class allowed us to detect significant potential for improvement in relation to effects on microtubules. Among other things, outstanding results have been obtained using the classical esterification of these borneolum acids is Sola substituted Skthe aim was to investigate the possibility to achieve high stability of microtubules by applying the compounds of this class compared to the use of Taxol.

Next, the following describes the compounds exhibiting pharmacological activity:

WO-A-9421252;

Bioorg. Med. Chem. Lett., (1994) 4(11), 1381-1384;

EP-A-0534708;

Angew. Chem. Int. Ed. Engl., (1994), 33(1), 15-44;

WO-A-9513053;

WO-A-9519994;

J. Pharm. Sci., (1995), 84(10), 1223-1230;

WO-A-9600724.

However, compounds described in WO-A-9421252 are the most loved ones in this field.

It has been unexpectedly discovered that compounds of the formula I according to this invention in comparison with Taxol and compounds known in the field, have a differentiated profile activity. Besides the obvious increased stability of microtubules, the compounds of formula I have additional effects on the polymerization of tubulin.

Taxanes according to this invention are characterized by the General formula I.

< / BR>
where Skcan be a

< / BR>
R1can represent hydrogen or C1-C10-acyl;

R2may represent-OH,

R3can be a C3or-CN; and

available hydroxy-group in the formula I can be further functionally modified by esterification or esterification,

as well as them, and cyclodextrine clathrates, as well as compounds of General formula I, enclosed in liposomes.

Under alkyl group, R3mean alkyl groups with unbranched or branched chain, containing from 1 to 10 carbon atoms, such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert.-butyl, pentyl, isopentyl, neopentyl, heptyl, hexyl and decyl. Preferred are alkyl groups containing from 1 to 4 carbon atoms. Acyl and alkoxygroup contained in R1and R3, respectively, of the General formula I, contain from 1 to 10 carbon atoms, and preferred are methoxy, ethoxy-, propoxy-, isopropoxy - and tert.-butoxypropyl, respectively.

Halogen in the symbol X represents fluorine, chlorine, bromine or iodine.

Preferred compounds of General formula I are:

3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonylamino,

3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxy,

3'-destiny-3'-(4-pyridyl)-2, educationsal.

The invention relates also to a method for borneol derivatives of the formula I, and this method is characterized by the fact that the alcohol of General formula II

< / BR>
where R1and R2defined here above, and hydroxy-group contained in the formula II, optionally protected, interacts with the compound of General formula IIIa, IIIb or S

< / BR>
< / BR>
< / BR>
where R3defined here above, and X' may represent a hydroxy, O-alkyl or halogen, and where available the hydroxy-group is protected by esterification or esterification, to obtain the compounds of General formula I, in which free hydroxy-group can be further functionally modified by esterification esterification.

In order to implement the esterification of an alcohol functional group in the compound II conduct deprotonation with base, such as, for example, a metal hydride (e.g. sodium hydride), an alcoholate of an alkali metal (for example, methanolate sodium tert-butanolate potassium), hexamethyldisilazane alkali metal (for example, hexamethyldisilazane sodium), 1,5-diazabicyclo [4.3.0 non-5-ene (DBN), 1,8-diazabicyclo [5.4.0]undec-7-ene (DBU), triethylamine, 4-(dimethylamino)-pyridine (DMAP) or 1,4-diazabicyclo[2.2.2] octm as, for example, dichloromethane, diethyl ether or tetrahydrofuran, at a temperature of from -70oC to +50oC. Preferred is the reaction with sodium hydride as the base, cyclic amidon acid as a carboxylic acid derivative and Tetra-hydrofuran as solvent at a temperature of from -10oC to +25oC.

Available hydroxy-group in the compound of the formula I can be further functionally modified in accordance with techniques known to experts in this field, for example by etherification or esterification. For example, the free hydroxy-group can be transferred in the form of pyridinium salts with physiologically tolerated acids, in the form of phosphates and their salts with physiologically tolerated bases or in the form of their esters or in the form of simple and complex esters of water-soluble polymers. Can also be obtained ethers and esters of the compounds, which themselves are able to exert inhibitory effects on the tumor.

Biological effects and scope of new derivatives of Taxol.

The new compounds of formula I are valuable drugs. They affect tubulin by vesatile to provide phasespecific effect on cell division. This impact is particularly strong impact on the rapidly growing tumor cells, the growth of which is significantly influenced by intracellular regulatory mechanisms. The active ingredients of this type are, in principle, are suitable for the treatment of diseases which can be therapeutic indications for the influence on the process of cell division.

As example can be mentioned the treatment of malignant tumors, malaria, treatment of diseases caused by gram-negative bacteria, and treatment of diseases of the Central and peripheral nervous system, which are based on excitotoxic mechanisms, such as, for example, the treatment of acute neurodegenerative conditions such as increased intracranial pressure (arise), for example as a result of stroke or traumatic brain injury, chronic neurodegenerative diseases, including Alzheimer's disease, and treatment of amyotrophic lateral sclerosis.

As examples of application in the treatment of malignant tumors may be mentioned, for example, used in the treatment of malignant tumors of the ovary, stomach, colon, prostate, mo is SS="ptx2">

In General, the compounds of this invention can be applied either by themselves or in order to achieve additive or synergistic effect in combination with other active principles and classes of compounds that can be applied in the required course of treatment.

When considering the treatment of tumors as example can be mentioned combinations with

the platinum complexes such as cisplatin and carboplatin,

penetrating compounds, for example, from the class anthracyclines, such as, for example, doxorubicin, or from the class of intrapersonal, such as, for example, C1-941,

compounds that act on tubulin, for example from the class of Vinca alkaloids, such as vincristine and vinblastine, or from a new class of esters of borneol, described in P 4416374.6 and 19513040.5, or from the class of macrolides, such as, for example, rhizoxin, or with other compounds, such as, for example, colchicine, combretastatin a-4 and epothilone and

inhibitors of DNA topoisomerases, such as, for example, camptothecin, etoposide, topotecan and teniposide,

folate or the pyrimidine antimetabolites, such as, for example, lometrexol and gemcitabin,

alkylating DNA hibitory PDGF, EGF, TGFb, EGF), such as, for example, antagonists of somatostatin,. (suramin), bombezin,

inhibitors tyrosinemia protein kinase or protein kinases A and C, such as, for example, erbstein, genistein, staurosporine, ilmofosine and 8-C1-camp,

antihormone means of class antigestagen means, such as, for example, mifepristone, onapristone, or from a class antiestrogenic means, such as, for example, tamoxifen, or from a class antiandrogenna means, such as, for example, tsiproteronatsetat,

compounds that inhibit metastasis, for example from a class of eicosanoids, such as, for example, PGI2, PGE1that 6-oxo-PGE1and their stable derivatives (for example, iloprost, cicaprost, beraprost),

the antagonists of Ca2+such as, for example, verapamil, gallopamil, flunarizin, diltiazem, nifedipine, and nimodipine,

neuroleptic means, such as, for example, chlorpromazine, trifluoperazine, thioridazine and perphenazine

local anesthetic means, such as, for example, carbanilate-Ca7cinchocaine, carbanilate-Ca3articaine, carbanilate, lidocaine,

compounds that inhibit the growth of blood vessels, such as, for example, anti-VEGF antibodies, endostatin IN,

This invention relates also to medicines on the basis of the compounds of General formula I, which are pharmaceutically acceptable, which means that the compounds are non-toxic in the concentrations, optionally combined with conventional excipients, carriers and additives.

Compounds according to this invention can be processed in accordance with the methods galenical pharmacy, known in themselves, in pharmaceutical preparations for enteral, percutaneous, parenteral or topical application. They can be applied in the form of tablets, pills, gelatin capsules, granules, suppositories, implants, sterile water and oil-based injectable solutions, suspensions or emulsions, ointments, creams and gels. Thus, this invention relates also to the use of compounds according to this invention in obtaining medicinal drugs.

Active(s) ingredient(s) may be mixed with fillers, which are adopted in galenical pharmacy, such as, for example, gum Arabic, talc, starch, mannitol, methylcellulose, lactose, surfactants such as Tweens or Myrj, magnesium stearate, aqueous or non-aqueous media, pria taste (for example, essential oils).

Thus, this invention relates also to pharmaceutical compositions and drugs, which as active constituent contain at least one compound according to this invention. A single dose contains from about 0.1 to 100 mg of active(s) ingredient(s). The dosage of the compounds according to this invention in humans of approximately from 0.1 to 1000 mg per day.

Description of the figures.

In Fig. 1 shows the changes of the absorption as a function of time and temperature (3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N - methoxycarbonylamino, applied example 2).

In Fig. 2 shows the changes of the absorption as a function of time and temperature (3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl - 3'-N-methoxycarbonyl-7-epitaxy, application example 3).

In Fig. 3 shows the changes of the absorption as a function of time and temperature (3'-destiny-3'-(4-pyridyl)-2,3'-N-misdemenor-3'-N - methoxycarbonylamino, applied example 4).

In Fig. 4 shows the changes of the absorption as a function of time and temperature (3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-10 - deacetyltaxol, applied example 5).

The following examples 1.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N - methoxycarbonylamino.

8,6 μl of 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran are added during 3oC in an atmosphere of dry argon to a solution of 3.1 mg (2.9 mmol) of the compound obtained according to example 1a in 0.5 ml of anhydrous tetrahydrofuran and the reaction mixture is stirred for 30 minutes. The mixture is then poured into a saturated solution of sodium bicarbonate and the extraction is carried out by adding dichloromethane, the organic extract was concentrated and the residue purified by chromatography method on the half plates for analytical thin-layer chromatography. As the mobile phase used ethyl acetate as eluent using a mixture of dichloromethane and methanol. Allocate 0.4 mg (0.5 mmol, 17%) specified in the connection header.

1H-NMR (CDCl3): d = 1.16 (3H), 1.25 (3H), 1.70 (3H), 1.75 (1H), 1.84 (3H), 1.90 (1H), 2.26 (3H), 2.25-2.38 (2H), 2.38 (3H), 2.48 (1H), 2.56 (1H), 3.62 (3H), 3.81 (1H). 4.19 (1H), 4.31 (1H), 4.40 (1H), 4.71 (1H), 4.95 (1H), 5.37 (1H), 5.66 (1H), 5.69 (1H), 6.28 (1H), 6.31 (1H), 7.40 (2H), 7.51 (2H), 7.61 (1H), 8.11 (2H), 8.66 (2H) ppm.

Example 1a.

2'-Triisopropylsilyl-3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-triethylsilanol.

12 mg approximately 60% dispersion of sodium hydride domestie with examples 1B and 1C, 0.1 ml of anhydrous tetrahydrofuran and the mixture is heated to 30oC and stirred for 30 minutes. The mixture is again cooled to 3oC add 30% aqueous acetic acid and several times the extraction is carried out by adding diethyl ether. The combined organic extracts washed with a saturated solution of sodium chloride and dried over magnesium sulfate. After filtration and removal of solvent the residue is purified by chromatography method on two plates for analytical thin-layer chromatography. As the mobile phase used a mixture of n-hexane and ethyl acetate as eluent using a mixture of dichloromethane and methanol. Allocate 3.7 mg (3.4 mmol, 57%) specified in the connection header.

1H-NMR (CDCl3): d = 0.60 (6H), 0.80-1.02 (30H), 1.25 (6H), 1.70 (3H), 1.91 (1H), 2.03 (3H), 2.14 (1H), 2.20 (3H), 2.36 (1H), 2.49 (3H), 2.53 (1H), 3.54 (3H), 3.84 (1H), 4.18 (1H), 4.30 (1H), 4.49 (1H). 4.85 (1H), 4.93 (1H), 5.30 (1H), 5.60 (1H), 5.70 (1H), 6.32 (1H), 6.47 (1H), 7.28 (2H), 7.49 (2H), 7.59 (1H), 8.13 (2H), 8.64 (2H) ppm.

Example 1B.

7-Triethylcitrate III

21 μl of Triethylchlorosilane and 10.3 mg of imidazole was added whenoC in an atmosphere of dry argon to a solution of 3.7 mg (6.3 mmol) of chromatographically purified baccatin III (Calbiochem Corp.) in 0.3 ml of anhydrous dimethylformamide and the reaction with the times the extraction is carried out by adding diethyl ether, followed by washing with a saturated solution of sodium chloride and the combined organic extracts are concentrated. The residue obtained after filtration and removal of the solvent, purified by the method of chromatography on half of the plate for analytical thin-layer chromatography. As the mobile phase used a mixture of n-hexane and ethyl acetate as eluent using a mixture of dichloromethane and methanol. Allocate 3.0 mg (5.6 mmol, 88%) specified in the connection header.

1H-NMR (CDCl3): d = 0.59 (6H),. 0.92 (9H), 1.06 (3H), 1.20 (3H), 1.62 (1H), 1.69 (3H), 1.88 (1H), 2.04 (1H), 2.19 (6H), 2.28 (2H), 2.29 (3H). 2.53 (IH). 3.88 (1H), 4.14 (1H), 4.31 (1H), 4.50 (1H), 4.83 (1H). 4.98 (IH). 5.63 (1H), 6.47 (1H), 7.49 (2H), 7.61 (1H), 8.11 (2H) ppm.

Example 1B.

(3R, 4S)-1-(Methoxycarbonyl)-3-triisopropylsilyl-4-(4-pyridyl)-2-azetidinone.

573 mg of 4-dimethylaminopyridine and 193 μl of methyl ether of Harborview acids are added at the 3oC in an atmosphere of dry argon to a solution of 250 mg (0.78 mmol) of the compound obtained according to example 1 g in 10 ml of anhydrous dichloromethane and the reaction mixture is heated to 23oC and stirred for another 16 hours. The mixture is then poured into a saturated solution of ammonium chloride, several times the extraction is carried out by adding diethyl ether, followed by washing with a saturated solution of sodium chloride and the combined organic extracts koi approximately 150 ml of fine silica gel, using as mobile phase a mixture of n-hexane and ethyl acetate. Allocate 251 mg (0.66 mmol, 85%) specified in the connection header.

1H-NMR (CDCl3): d = 0.82-1.07 (21H), 3.82 (3H), 5.11 (1H), 5.26 (1H), 7.23 (2H), 8.61 (2H) ppm.

Example g

(3R,4S)-3-Triisopropylsilyl-4-(4-pyridyl)-2-azetidinone

The solution to 67.3 g of cerium ammonium nitrate in 700 ml of water is added at the 3oC in an argon atmosphere to a solution of 17.2 g (of 40.3 mmol) of the compound obtained according to example 1D, in 384 ml of anhydrous acetonitrile and the reaction mixture is stirred for 30 minutes. The mixture is then poured into a saturated solution of sodium bicarbonate, several times the extraction is carried out by adding ethyl acetate and the combined organic extracts are washed with 1% sodium hydroxide solution and dried over magnesium sulfate. The residue obtained after filtration and removal of the solvent, purified by the method of chromatography on approximately 800 ml of fine silica gel, using as mobile phase a mixture of n-hexane and ethyl acetate. Allocate 7,89 g (24.6 mmol, 61%) specified in the connection header.

1H-NMR (CDCl3): d = 0.78-1.07 (21H), 4.81 (1H), 5.23 (1H), 6.39 (1H), 7.28 (2H), 8.59 (2H) ppm.

Example 1D.

(3R, 4S)-1-(4-Methoxyphenyl)-3-is this Diisopropylamine in 70 ml of anhydrous tetrahydrofuran is cooled to -30oC in an atmosphere of dry argon, add to 37.6 ml of a 2.4 M solution of n-utility in n-hexane and the mixture is heated to 0oC. After 30 minutes the mixture is cooled to -78oC, it is added dropwise a solution of 22.1 g (56,6 mmol) (1R, 2S)-2-phenyl-1-cyclohexyltrichlorosilane obtained in accordance with methods similar to those described in Tetrahedron, T. 48, No. 34, page 6985-7012, 1992, in 70 ml of anhydrous tetrahydrofuran and the mixture is stirred for 3 hours. Then add a solution of 15.6 g (of 73.5 mmol) aldimine obtained in accordance with example 1, in 150 ml of anhydrous tetrahydrofuran and the mixture is heated to 23oC for 16 hours. The mixture is then poured into a saturated solution of ammonium chloride, several times the extraction is carried out by adding ethyl acetate followed by washing with a saturated solution of sodium chloride and the combined organic extracts are concentrated. The residue obtained after filtration and removal of the solvent, purified by the method of chromatography on approximately 1.8 liters porous silica gel, using as mobile phase a mixture of n-hexane and ethyl acetate. Allot of 17.2 g (of 40.3 mmol, 71%) specified in the connection header.

1H-NMR(CDCl3): d = 0.82-1.12 (21H), 3.76 (3H), 5.12 (1H), 5.29 (1H), 6.80 (2H), 7.19-4-aldehyde and 8.4 g of magnesium sulfate is added in an atmosphere of dry argon to a solution of 10 g (81,1 mmol) of 4-anisidine in 120 ml of anhydrous dichloromethane and the mixture was stirred at 23oC for 4 hours. The residue obtained after filtration and removal of solvent, recrystallized from n-hexane. Allocate 15.9 g (74.9 mmol, 92%) specified in the connection header.

1H-NMR (CDCl3): d = 3.83 (3H), 6.95 (2H), 7.29 (2H), 7.73 (2H), 8.47 (1H), 8.73 (2H) ppm.

Example 2.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxial (A) and 3'-destiny-3'-(4-pyridyl)-2,3'-N-bis-Dibenzoyl-3'-N - methoxycarbonylamino (B)

42 μl of 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran are added during 3oC in an atmosphere of dry argon to a solution of 15 mg (a 13.9 mmol) of the compound obtained according to example 1A in 0.5 ml of anhydrous tetrahydrofuran and the reaction mixture is stirred for 30 minutes at 3oC, heated up to 23oC and stirred for another 30 minutes. The mixture is then poured into a saturated solution of sodium bicarbonate, the extraction is carried out by adding dichloromethane, the organic extract was concentrated and the residue purified by chromatography method on two plates for analytical thin-layer chromatography. As the mobile phase used a mixture of ethyl acetate and methanol as eluent using a mixture of dichloromethane and methanol.

Wydania B, and 1.2 mg (1.5 mmol, 11%) of the compound described in example 1.

1H-NMR (CDCl3) of A: d = 1.18 (3H), 1.23 (3H), 1.68 (3H), 1.71 (1H), 1.80 (1H), 1.83 (3H), 2.15-2.48 (4H), 2.21 (3H), 2.49 (3H), 3.56 (3H), 3.71 (1H), 3.92 (1H), 4.37 (2H), 4.63 (1H), 4.71 (1H), 4.91 (1H), 5.37 (1H), 5.67 (1H), 5.76 (1H), 6.34 (1H), 6.81 (1H), 7.33 (2H), 7.51 (2H), 7.61 (1H), 8.16 (2H), 8.63 (2H) ppm.

Example 3.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N - methoxycarbonyl-10-deacetyltaxol.

20,4 ál 4 N. hydrochloric acid is added in an atmosphere of dry argon to a solution of 9.0 mg (10.2 mmol) of the compound As obtained in accordance with example 3A, 0.8 ml of ethanol and 0.2 ml of tetrahydrofuran and the reaction mixture stirred at 23oC for one hour. Adding hydrochloric acid repeat twice more, each time after the one-hour period of stirring, add a saturated solution of sodium bicarbonate, the extraction is carried out by adding dichloromethane, the organic extract was concentrated and the residue purified by chromatography method on two plates for analytical thin-layer chromatography. As the mobile phase used a mixture of ethyl acetate and ethanol as eluent using a mixture of dichloromethane and methanol. Allot of 6.5 mg (8.5 mmol, 83%) specified in the connection header.

1(1H), 5.21 (1H), 5.36 (1H), 5.68 (1H), 5.83 (1H), 6.30 (1H), 7.34 (2H), 7.50 (2H), 7.62 (1H), 8.10 (2H), 8.61 (2H) ppm.

Example 3A.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N - methoxycarbonyl-7-triethylsilyl-10-deacetyltaxol (A) and

3'-Destiny-3'-(4-pyridyl)-2,3'-N-bis-Dibenzoyl-3' -N-methoxycarbonyl-7-triethylsilyl-10-deacetyltaxol (B).

Conduct the reaction 25 mg (23 μmol) of crude product obtained in accordance with example 3b, if -10oC, similar to that of example 1, and after cleaning and allocate 9.0 mg (10.2 mmol, 44%) specified in the connection header And 2.5 mg (3.2 mmol, 14%) specified in the connection header B, and 2.2 mg (2.9 mmol, 12%) specified in the title compound from example 3.

1H-NMR (CDCl3) of A: d = 0.43-0.67 (6H), 0.94 (9H), 1.13 (3H), 1.24 (3H), 1.70 (1H), 1.76 (3H), 1.87 (3H), 1.93 (1H), 2.24 (2H), 2.36 (3H), 2.48 (1H), 3.62 (3H), 3.87 (1H), 4.18 (1H), 4.29 (1H), 4.34 (1H), 4.68 (1H), 4.91 (1H), 5.12 (1H), 5.36 (1H), 5.63 (1H), 5.77 (1H), 6.29 (1H), 7.33 (2H), 7.49 (2H), 7.60 (1H), 8.10 (2H), 8.60 (2H) ppm.

1H-NMR (CDCl3) of: d = 0.45-0.64 (6H), 0.94 (9H), 1.02 (3H), 1.25 (4H), 1.67 (1H), 1.72 (3H), 1.82 (3H), 1.88-2.12 (2H), 2.18 (4H), 2.47 (1H), 3.50 (1H), 3.67 (1H), 3.70 (3H), 3.90 (1H), 4.26 (1H), 4.30 (1H), 4.59 (1H), 4.61 (1H), 4.66 (1H), 4.91 (1H), 5.03 (1H), 5.27 (1H), 5.65 (1H), 6.25 (1H), 7.32 (2H), at 8.62 (2H) ppm.

Example 3b.

2'-Triisopropylsilyl-3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-triatlo 25 mg (23.2 mmol) of the compound, obtained in accordance with example 1A, 1.2 ml of ethanol and the mixture was stirred at 23oC for 24 hours. The mixture is then poured into a saturated solution of ammonium chloride, several times the extraction is carried out by adding ethyl acetate, and drying carried out over magnesium sulfate. The residue obtained after filtration and removal of solvent used in further reactions without purification. Allocate 22 mg (max 21 µm, Max. 91%) indicated in the title compound, which still contains a small amount of the original product.

The following application examples illustrate the biological activity of the compounds according to this invention without limiting their practical application in these examples.

Application example 1.

Isolation and purification of tubulin.

From directly in front of this dissected cow skulls remove the brain of the cow (each weighing 330 g) and transported in ice-cold buffer RM) Each brain freed from meninges and all clots and homogenized sufficient buffer RM-M in the cold room. The homogenate 2 cow head brain brought to a total volume of 1.0 liter of the total quantity of 500 ml buffer and subjected to pervert through 4 layers of thin muslin, carry in a balanced centrifuge tubes (420 ml) and centrifuged again (Ti rotor 45, 96000 g, 75 minutes, 4oC). The supernatant is separated from the sediment using a pipette and filtered through 6 layers of thin muslin, and add 50 mm GTP solution in 0.01 M bicarbonate buffer/FBI to obtain a final concentration of 1 mm. Initially, the polymerization is carried out in balanced, centrifuge the tubes for 45 minutes in a water bath heated to 37oC. the Resulting microtubules allocate by centrifugation (rotor Ti 45, 27oC, 96000 g, 60 minutes), the supernatant carefully removed with a pipette and very delicate opalescent precipitate carefully separated from the wall with a spatula. Then to the precipitate add 40 ml of cold buffer RM followed by homogenization using a small glass homogenizer-mortar and incubation overnight (12 to 16 hours) on ice in a balanced centrifuge tubes in a cold room. The depolymerization product is removed by centrifugation at Ti rotor 60 (4oC, 96000 g, 60 minutes) and the supernatant was diluted with buffer RM-M in a 1:1 ratio, incubated in a balanced centrifuge the tubes for 45 minutes and centrifuged again (Ti rotor 45, 27oC, 96000 g, 60 minutes). With the l cold buffer RM, gently homogenized using a glass homogenizer-mortar and incubated on ice for 30 minutes. By repeated centrifugation (Ti rotor 60, 4oC, 96000 g, 60 minutes) get the drug tubulin, the protein content of which is determined by Pearce (Pearce) or by photometric measurement at 280 nm. When determining the protein concentration used in the cultivation of the selected material in the buffer RM 1:10, 1:20 and 1:40. The RM buffer has its own value of extinction and is deducted as a zero value of the defined value of the protein content. The selected material is diluted with a buffer of the Republic of Moldova to the desired protein concentration (2 mg/ml).

Applied example 2.

Biological effects of 3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonylamino on tubulin.

Measurement of tubulin polymerization and depolymerization of microtubules carried out according to the method of photometry. Before measuring the drug tubulin obtained in accordance with application example 1, thawed and Tegaserod within 15 minutes. Photometer set at a wavelength of 350 nm. 3 μl of the solution/sample, 6 μl of GTP (final concentration 0-25 mmol/l) and 291 μl of drug tubulin (2 mg protein/ml) are selected using PIP is slowly placed into the cell holder and the measurement procedure starts at 37oC. as soon As the maximum level of polymerization (control and solvent and Taxol 1E-5 mol/l after 20 minutes), stimulate the beginning of depolymerization by lowering the temperature to 15oC. measurement Procedure terminate upon completion of the depolymerization process and the changes of absorption present in graphical form as a function of time and temperature (Fig. 1).

From the figure it is clear that the Taxol accelerates the polymerization of tubulin in comparison with control and inhibits depolymerization, whereas the connection according to this invention, the 3'-destiny-3'-(4-pyridyl)-3'-N - desbenzoyl-3'-N-methoxycarbonylamino inhibits polymerization and stabilizes the resulting microtubules are noticeably better than Taxol.

Application example 3.

Biological effects of 3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxy on tubulin.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N - methoxycarbonyl-7-epitaxial causes a much more pronounced acceleration of the polymerization of tubulin and stabilizes the resulting microtubules are noticeably better than Taxol. The results are shown in Fig. 2.

Application example 4.

Biological effects of 3'-destinies-Dibenzoyl-3'-N-methoxycarbonylamino causes a much more pronounced acceleration of the polymerization of tubulin and stabilizes the resulting microtubules better than Taxol. The results are shown in Fig. 3.

Application example 5.

Biological effects of 3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-10-deacetyltaxol on tubulin.

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-10-deacetyltaxol not differ from Taxol in terms of effects on the polymerization of tubulin, but it stabilizes the formed microtubules significantly better than Taxol. The results are shown in Fig. 4.

Application example 6.

Antiproliferative effect on tumor cell lines:

3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonylamino (1),

3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxial (2),

3'-destiny-3'-(4-pyridyl)-2,3'-N-bis-Dibenzoyl-3'-N-methoxycarbonylamino (3) and

3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-10-deacetyltaxol (4).

Cell lines of breast carcinoma MDA MB 435 (5000 cells/well) put on tablets for micrometrology (day 0, medium RPMI, 1% nonessential amino acids, 1% pyruvate, 10% amniotic calf serum). 1 day spend adding connections in several concentrations. Antiproliferative effect estimate is">

3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonylamino shows activity similar to that of Taxol, whereas the 3'-destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxial shows significantly enhanced inhibitory activity compared with Taxol.

1. Taxanes General formula I, with pharmacological activity,

< / BR>
where Skcan be a

< / BR>
R1can represent hydrogen or C1- C10-acyl;

R2can be a HE;

R3can be a C1- C10-alkoxygroup,

and the free hydroxy-group in the I can be further functionally modified by esterification.

2. 3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonylamino under item 1.

3. 3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-7-epitaxy under item 1.

4. 3'-Destiny-3'-(4-pyridyl)-2,3'-N-bis-Dibenzoyl-3'-N-methoxycarbonylamino under item 1.

5. 3'-Destiny-3'-(4-pyridyl)-3'-N-desbenzoyl-3'-N-methoxycarbonyl-10-deacetyltaxol under item 1.

6. The method of obtaining taxonomic derivatives of General formula I on PP.1 to 5, characterized in that the alcohol of General formula II

< / BR>
DG is Bered, subjected to interaction with the compound of General formula IIIa, IIIb or IIIc

< / BR>
< / BR>
< / BR>
where R3has a specified value;

X' may represent a hydroxy, O-alkyl or halogen,

and where the free hydroxy-group is protected by esterification or esterification, to obtain the compounds of General formula I, in which free hydroxy-group can be further functionally modified by esterification.

7. Drug, possess cytotoxic activity containing one or more compounds of formula I on PP.1 to 5, is able to inhibit polymerization and stabilize the formed microtubules.

8. Drug under item 7, additionally including traditional fillers, carriers and additives.

 

Same patents:

-alaninemia and their derivatives, method for the treatment glycolipoprotein diseases, the prevention of ischemic myocardial damage, the pharmaceutical composition" target="_blank">

The invention relates to derivatives of indole-2-carboxamide that can be used as inhibitors of glycogen phosphorylase, and to methods of treatment of glycogenolysis-dependent diseases or conditions using these compounds and pharmaceutical compositions containing these compounds

The invention relates to new derivatives of chromone General formula 1, in which ring a is unsubstituted or one-deputizing halogen, and where the ring is unsubstituted or substituted by one to four substituents selected from the group consisting of lower alkyl, hydroxyl, lower alkoxyl, lower alkylthio or halogen, and their salts, also describes a method of production thereof and pharmaceutical composition based on compounds of the formula I, which has antagonistic activity against neirokinina 1

The invention relates to new derivatives of amino(thio)ethers of the formula I

< / BR>
where X represents oxygen, sulfur, sulfinil, sulfonyl or, if R0and R1together are not alkalinous chain with 1 to 3 atoms, CH2:

Z represents -(CH2)n1-(CHA)n2-(CH2)n3and

n1 = 0, 1, 2 or 3,

n2 = 0 or 1,

n3 = 0, 1, 3 or 3, provided that

n1 + n2 + n3 < 4;

R0represents hydrogen or A;

R1represents hydrogen, A, OA, phenoxy, Ph, OH, F, Cl, Br, CN, CF3, COOH, COOA, acyloxy with 1-4 carbon atoms, carboxamido, -CHNH2, -CH2NHA, -CH2NA2,

-CH2NHAc, -CH2NHSO2CH3,

or

R0and R1together represent alkylenes chain with 1 to 3 carbon atoms or alkenylamine chain with 2 to 3 carbon atoms;

R2represents hydrogen, A, Ac, or-CH2-R4;

R3represents-CH2-R4or-CHA-R4;

R4is a Ph, 2-, 3 - or 4-pyridyl (unsubstituted or monosubstituted R5) or thiophene (unsubstituted, mono - or disubstituted by A, OA, OH, F, Cl, Br, CN and/or CF3or the other what it is, di-, tri-, Tetra-, or pentamidine F, CF3partially or fully fluorinated A, A and/or OA;

R6, R7, R8and R9each independently represents H, A, OA, phenoxy, OH, F, Cl, Br, I, CN, CF3, NO2, NH2, NHA, NA2, Ac, Ph, cycloalkyl c 3-7 carbon atoms, -CH2NH2, -CH2NHA, -CH2NA2, -CH2N HAC or-CH2NHSO2CH3or two coming together constitute the remainder alkylenes chain with 3-4 carbon atoms, and/or R1and R6together predstavljaet a chain with 3 or 4 carbon atoms;

A represents alkyl with 1-6 carbon atoms;

Ac is alkanoyl with 1-10 carbon atoms or aroyl with 7 to 11 carbon atoms;

Ph represents phenyl (unsubstituted or substituted R5, 2-, 3 - or 4-pyridium or phenoxyl group);

and physiologically acceptable salts, their derivatives

The invention relates to new derivatives of phenyl-oxo-alkyl-(4-piperidinyl)benzoate of formula I, their N-oxide forms, salts and steric isomer forms, where R1- halogen, R2is hydrogen, R3- C1-6-alkyl, or R2and R3form together a bivalent radical of the formula -(CH2)2- or -(CH2)3-, Alk-C1-6-alcander, R4is hydrogen or C1-6-alkoxy, R5, R6and R7is hydrogen, halogen, C1-6-alkyl, C1-6-alkyloxy or R5and R6taken together , form a bivalent radical of the formula-NR8-C(O)-NR9- or-NH-C(NH-R10)=N-, where R8and R9is hydrogen, C1-6-alkyl, R10is hydrogen, C1-6-alkylsulphonyl, C1-6-allyloxycarbonyl

The invention relates to new N-substituted piperidinylmethyl f-ly I, their N-oxide forms, isomers, and salts, where R1- halogen, C1-6alkylsulfonamides And divalent radical-CH2-CH2; -CH2-CH2-CH2- or-CH=CH-; R2is hydrogen or C1-6alkyloxy; L is a radical of formula-Alk-R4, -Alk-OR5, -Alk-NR6R7; Alk-C1-12alcander; R4is hydrogen, cyano, C1-6alkylsulphonyl,1-6allyloxycarbonyl, etc

The invention relates to a derivative of 7- (alkoxycarbonyl-substituted) -10-hydroxy-taxan following formula 3b:

< / BR>
in which R1, R2, R4, R5, R6and R14defined above

The invention relates to new derivatives of 10-deazetil-14-hydroxyacetone III of formula 1, where X represents a group >C=S, >C=NH or >S=O; OR1which can beororiented, represents hydroxy, alkylsilane (preferably triethylsilane, O-TES); R2isororiented hydroxy group or Troc group (Troc= Cl3CCH2COO-), or with the carbon atom to which it is attached, forms ketogroup; R3is izoterikoy residue of formula 2; R4is a linear or branched alkyl or alkenylphenol group having 1 to 5 carbon atoms; R5is alkyl having 1 to 5 carbon atoms or tert-butoxypropyl

The invention relates to new intermediate products for poluentes taxan and methods for their preparation

The invention relates to new taxoids General formula (I)

< / BR>
in which Rameans a hydrogen atom or a hydroxyl radical, CNS radical with 1-4 carbon atoms, acyloxy radical with 1-4 carbon atoms or alkoxyacetic-radical, the alkyl part of which contains 1-4 carbon atoms;

Rbmeans a hydrogen atom; or

Raand Rbtogether with the carbon atom to which they are bound, form a ketone function;

Z means a hydrogen atom or a radical of General formula (II)

< / BR>
in which R1means bentely radical, possibly substituted by one or more identical or different atoms or radicals selected among halogen atoms and alkyl radicals with 1-4 carbon atoms, CNS radicals with 1-4 carbon atoms or trifloromethyl; tenor or furoyl, or the radical R2-O-CO-, in which R2means:

is an alkyl radical with 1-8 carbon atoms, alkanniny radical with 2-8 carbon atoms, alkynylaryl radical with 3-8 carbon atoms, cycloalkenyl radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 atoms in or more substituents, selected among halogen atoms and hydroxyl radical, CNS radical with 1-4 carbon atoms, dialkylamino radical, each alkyl portion of which contains 1-4 carbon atoms, piperidino radical, morpholino radical, piperazine-1-ilen radical (possibly substituted in position 4 alkyl radical with 1-4 carbon atoms or phenylalkyl radical, the alkyl part of which contains 1-4 carbon atoms), cycloalkyl radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 carbon atoms, phenyl radical (possibly substituted by one or more atoms or radicals, selected among halogen atoms and alkyl radicals with 1-4 carbon atoms or CNS radicals with 1-4 carbon atoms), a cyano radical, carboxyl or alkoxycarbonyl, the alkyl portion of which contains 1-4 carbon atoms;

is phenyl or- or-nattily radical, possibly substituted by one or more atoms or radicals selected among halogen atoms and alkyl radicals with 1-4 carbon atoms or CNS radicals with 1-4 carbon atoms; or a heterocyclic aromatic patterollers radical with 4 to 6 carbon atoms, possibly substituted by one or more alkyl radicals with 1-4 carbon atoms;

R3means a linear or branched alkyl radical with 1-8 carbon atoms, a linear or branched alkanniny radical with 2-8 carbon atoms, linear or branched alkynylaryl radical with 2-8 carbon atoms, cycloalkyl radical of 3-6 carbon atoms, phenyl or- or-nattily radical, possibly substituted by one or more atoms or radicals selected among halogen atoms and alkyl, alkenyl, etkinlik, aryl, kalkilya, CNS, alkylthio, aryloxy, aaltio-, hydroxyl, hydroxyalkyl, mercapto-, formyl, acyl, acylamino, aroylamino, alkoxycarbonyl-, amino-,

alkylamino, dialkylamino-, carboxyl, alkoxycarbonyl, karamolegos, alkylcarboxylic, dialkylanilines, cyano-, nitro - and triptorelin radicals; or an aromatic five-membered a heterocycle containing one or more, identical or different heteroatoms selected among nitrogen atoms, oxygen or sulfur, and possibly substituted by one or more of the ylamino - dialkylamino-, alkoxycarbonyl-, acyl, arylcarboxylic, carboxyl, karamolegos, alkylcarboxylic, dialkylanilines or alkoxycarbonyl radicals, provided that the substituents phenyl,- or-afternova and aromatic heterocyclic radicals, the alkyl radicals and alkyl portions of other radicals contain 1-4 carbon atoms; alkeneamine and alkyline radicals contain 2 to 8 carbon atoms, and aryl radicals are phenyl or- or-raftiline radicals; and

R4means:

- linear or branched alkyl radical with 1-8 carbon atoms, a linear or branched alkanniny radical with 2-8 carbon atoms, linear or branched alkynylaryl radical with 2-8 carbon atoms, cycloalkyl radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 carbon atoms or bicycloalkyl radical with 7 to 11 carbon atoms, and each of these radicals may be substituted by one or more substituents selected among halogen atoms and hydroxyl radical, alkoxyl carbon piperidino radical, morpholino radical, piperazine-1-ilen radical (possibly substituted in position 4 alkyl radical with 1-4 carbon atoms or phenylalkyl radical, the alkyl part of which contains 1-4 carbon atoms), cycloalkyl radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 carbon atoms, possibly substituted phenyl radical, cyano radical, carboxyl or alkoxycarbonyl, the alkyl portion of which contains 1-4 carbon atoms;

- or aryl radical, possibly substituted by one or more atoms or radicals selected among halogen atoms and alkyl, alkenyl, etkinlik, aryl, kalkilya, CNS, alkylthio, aryloxy, aaltio-, hydroxyl, hydroxyalkyl, mercapto-, formyl, acyl, acylamino, aroylamino, alkoxycarbonyl-, amino-, alkylamino, dialkylamino-, carboxyl, alkoxycarbonyl, karamolegos, alkylcarboxylic, dialkylanilines, cyano-, nitro-, azido-, triptorelin or triptoreline radicals;

or saturated or unsaturated 4-6 membered heterocyclyl radical, possibly substituted by one or more alkyl radicals with 1-4 of carbon atoms is a branched alkanniny radical with 2-8 carbon atoms, linear or branched alkynylaryl radical with 2-8 carbon atoms, cycloalkyl radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 carbon atoms or bicycloalkyl radical with 7 to 11 carbon atoms, these radicals can be substituted by one or more substituents selected among halogen atoms and hydroxyl radical, CNS radical with 1-4 carbon atoms, dialkylamino radical, each alkyl portion of which contains 1-4 carbon atoms, piperidino radical, morpholino radical, piperazine-1-ilen radical (possibly substituted in position 4 alkyl radical with 1-4 carbon atoms or phenylalkyl radical, the alkyl portion of which contains 1-4 carbon atoms), cyanoaniline radical of 3-6 carbon atoms, cycloalkenyl radical with 4 to 6 carbon atoms, possibly substituted phenyl radical, cyano radical, carboxyl or alkoxycarbonyl, the alkyl portion of which contains 1-4 carbon atoms;

provided that cycloalkyl, cycloalkenyl or bicycloalkyl radicals can be substituted by one or more alkyl radicals with 1-4 carbon atoms

The invention relates to new taxoids General formula I, in which R denotes a linear or branched alkyl radical with 1-6 carbon atoms, Z denotes a hydrogen atom or a radical of General formula II, where R1means bentely radical or the radical R2-O-CO-, in which R2denotes an alkyl radical with 1-8 carbon atoms, R3denotes a phenyl radical

The invention relates to medicine and is a method of prevention and/or treatment of autoimmune diseases, including the introduction of an effective amount of pharmaceutically acceptable salt or complex of magnesium
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