Synthesis of epothiliones, intermediate products thereof, analogues and use thereof

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula or a pharmaceutically acceptable salt thereof, in which R1 denotes hydrogen or C1-6alkyl; R2 denotes isooxazolyl group, substituted with C1-6alkyl; RB denotes -CF3, -CHF2, -CH2F, or C1-6alkyl. The invention also relates to pharmaceutical compositions for treating cancer which contain the disclosed compounds.

EFFECT: obtaining novel compounds and a pharmaceutical compositions based on said compounds, which can be used in medicine for treating cancerous diseases.

15 cl, 77 dwg, 10 tbl, 13 ex

 

Epothilone A and B (2a and 2b, scheme 1) are naturally occurring cytotoxic macrolides, which have been isolated from destroying the cellulose Mycobacterium Sorangium cellulosum (Höfle et al. Angew. Chem., Int. Ed. Engl. 1996, 35, 1567, and J. Antibiot. 1996, 49, 560; each of the publications included in this description by reference). Despite the huge number of different structures epothilone A and B have a common mechanism of action, the same as the mechanism of action of paclitaxel (Taxol®), which is involved in the inhibition of growth of tumor cells in the polymerization of tubulin and stabilize the Assembly of microtubules (Bollag et al. Cancer Res. 1995, 55, 2325; incorporated by reference). Despite its indisputable clinical value as a chemotherapeutic front edge, Taxol® is far from ideal medicines. Its slight solubility in water makes it necessary to resort to the help of fillers to the composition, such as cremophor, which themselves pose risks and challenges when working with them (Essayan et al. J. Allergy Clin. Immunol. 1996, 97, 42; publication are included in this description by reference). In addition, Taxol® vulnerable to deactivation caused by multidrug resistance (MDR) (Giannakakou et al. J. Biol. Chem. 1997, 272, 17118; publication are included in this description by reference). However, it is also shown that epothilone A and B retain appreciable efficiency is Yunosti against MDR tumor cells (Kowalski et al. Mol. Biol. Cell 1995, 6, 2137; publication are included in this description by reference). In addition, increased water solubility compared with paclitaxel may be useful to be able to prepare compositions epothilones. While naturally occurring compound, epothilone B (2b, EpoB scheme 1) is an effective representative epothilone family of natural products, unfortunately, he has, at least in mice with xenografts of concern limited therapeutic index (Su et al. Angew. Chem. Int. Ed. Engl. 1997, 36, 1093; Harris et al. J Org. Chem. 1999, 64, 8434; each publication included in this description by reference).

Bearing in mind the limited therapeutic index EpoB, researched other analogues epothilone, in particular 12,13-desoxyepothilone, in respect of their ability to give an improved therapeutic profile (see U.S. patent№№ 6242469, 6284781, 6300355, 6369234, 6204388, 6316630; each of which is incorporated in this description by reference). The in vivo experiments conducted on various mouse models showed that the 12,13-desoxyepothilone B (3b, dEpoB figure 2) has therapeutic potential against various sensitive and resistant human tumors in xenografts of mice (Chou et al. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 9642 and 15798; publication are included in this description by reference). Weeks the VNO therapeutic advantage of these desoxyepothilone compared with other antitumor agents finally shown through comparative studies (Chou et al. Proc. Natl. Acad. Sci. U. S. A. 2001, 98, 8113; publication are included in this description by reference). Due to an impressive profile in vivo dEpoB was further subjected to Toxicological studies on dogs and is currently undergoing human trials as anticancer drugs.

In light of the promising therapeutic use 12,13-desoxyepothilone may be desirable to explore additional analogues, as well as additional methods of synthesis for the synthesis of existing epothilones, desoxyepothilone and their analogues, as well as their new counterparts. In particular, in the presence of interest in therapeutic use of this class of compounds may be desirable to develop techniques capable of providing significant amounts of any epothilones or desoxyepothilone previously described or described in this application for clinical trials and for large-scale retrieval.

Description of the drawings

Figure 1 represents a table of values IC50for epothilones in relation to cell growth CCRF-CEM, CCRF-CEM/VBL and CCRF-CEM/Taxol. Inhibition of cell growth was measured using analysis based on tetrazone Templ after 72-hour incubation for cell growth, as described previously (Scudiero et al. Cancer Res. 46: 4827-4833, 1988; publication are included in this description as SS is the CTL). The values of the IC50was determined on the basis of the relationship dose-effect at six or seven concentrations of each drug using a computer program (Chou et al. Adv. Enzyme Regul. 22: 27-55, 1984; Chou et al. CalcuSyn for Windows (Biosoft, Cambridge, UK), 1997; each of which is incorporated in this description by reference), as described previously (Chou et al. Proc. Natl. Acad. Sci. USA 95: 15798-15802, 1998; this publication is included in this description by reference).

Figure 2 is1H-NMR spectrum of TRANS-9,10-degidro-12,13-disoxaril.

Figure 3 is13C-NMR spectrum of TRANS-9,10-degidro-12,13-disoxaril.

Figure 4 shows the scheme for synthesis of 11-R - 14-R-epothilones using metathesis of olefins with snapping cycle LACDAC, and illustrates some of substitution available in the case of synthesis methods, which pass through 9,10-dehydroemetine.

Figure 5 presents data relative cytotoxicity against leukemic cells human in vitro for a number of compounds and derivatives epothilone, including some 9,10-dihydrocodeinone (for example, compound 7 on figa and connection 88 and 89 on figv).

Figure 6 depicts an alternative method of synthesis for the preparation of analogues 9,10-dehydroemetine. On figa shows the technique Macro-Steele, methodology mate sp3-sp3and methods β-Suzuki. On FIGU it is shown how referirovanija Julia methodology Wadsworth-Emmons and m the method of Macro-reformed. On figs it is shown how a combination of Macmurry and synthesis of an analogue of the lactam.

7 shows various analogues 9,10-degidro-12,13-disoxaril.

On Fig shows therapeutic effect of 9,10-degidro-dEpoB and dEpoB nude mice bearing xenograft breast carcinoma human MX-1 (in/in-infusion, Q2Dx3).

Figure 9 shows the stability analogues epothilone in the plasma of mice. Epo 1 means 12,13-disoxaril, Epo 2 means 26-F3-12,13-disoxaril, Epo 3 means (E)-9,10-degidro-12,13-disoxaril and Epo 4 means 26-F3-(E)-9,10-degidro-12,13-disoxaril.

Figure 10 depicts therapeutic effect of analogues epothilone in mice nude (naked), bearing xenograft HCT-116 (in/in-infusion, Q2Dx7, n=3). Arrows indicate the administration of a medicinal product. Epo 3 means (E)-9,10-degidro-12,13-disoxaril.

Figure 11 shows the effectiveness of various analogues epothilone against the growth of tumor cells in vitro and therapeutic index compared with paclitaxel and vinblastine.

Fig is a table summarizing the effect of dEpoB, Taxol and 26-F-9,10-deH-dEpoB against xenograft MX-1 nude mice.

On Fig shown a therapeutic effect on 26 trifter-9,10-degidro-dEpoB and 9,10-degidro-EpoB on the size of the tumors in nude mice bearing xenografts MX-1 (6 hours/in infusion, Q2Dx6 and Q2Dx9, respectively).

On Fig shows the changes in body weight of nude mice, bearing the appropriate xenograft tumor breast carcinoma human MX-1, after treatment 26 trifter-9,10-degidro-dEpoB and 9,10-degidro-EpoB (6-hour infusion, Q2Dx6 and Q2Dx9, respectively).

On Fig shown a therapeutic effect on 26 trifter-9,10-degidro-dEpoB and 9,10-dihydroiso on the size of the tumors in nude mice bearing xenografts MX-1 (6 hours/in infusion, Q2Dx6 and Q2Dx9, respectively).

On Fig shows the changes in body weight of nude mice bearing xenograft tumors of breast carcinoma human MX-1, after treatment 26 trifter-9,10-degidro-dEpoB and 9,10-degidro-EpoB (6 hours/in infusion, Q2Dx6 and Q2Dx9, respectively).

On Fig shows therapeutic effect of 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts HCT-116 (in/in-infusion, Q2Dx7).

On Fig shows the effect of 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts of carcinoma of the colon of human HCT-116 (in/in-infusion, Q3Dx5).

On Fig shows the effect of 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts A549/Taxol (6 hours/in infusion, Q3Dx7).

On Fig shows the changes in body weight of nude mice bearing xenograft A549/Taxol-treated 26 trifter-9,10-degidro-dEpoB and 9,10-degidro-dEpoB (6 hours/in infusion, Q3Dx7).

On Fig shows the effect of 26-Cryptor-9,10-degidro-dEpoB and 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts A549/Taxol (6 hours/in infusion, Q2Dx7).

On pig show what s the changes in body weight of nude mice, bearing xenografts A549/Taxol-treated 26 trifter-9,10-degidro-dEpoB and 9,10-degidro-dEpoB (6 hours/in infusion, Q2Dx7).

On Fig shows the effect of 9,10-degidro-EpoB on the size of the tumors in nude mice bearing xenografts of tumor HCT-116 carcinoma of the colon of a person (6 hours/in infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft tumors of HCT-116 colon carcinoma intestine of man, after treatment with 9,10-degidro-EpoB (6 hours/in infusion).

On Fig shows the formation of microtubules from tubulin in the presence of various analogues epothilone at 37°C.

On Fig shows the formation of microtubules from tubulin in the presence of various analogues epothilone at 4°C.

On Fig shows the effect of 9,10-degidro-dEpoB and dEpoB on the size of the tumors in nude mice bearing xenografts HCT-116 (in/in-infusion, Q2Dx6).

On Fig shows the changes in body weight of nude mice bearing xenografts HCT-116, after treatment with 9,10-degidro-dEpoB and dEpoB (in/in-infusion, Q2Dx6).

On Fig shows the effect of 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts HCT-116 carcinoma of the colon of a person (in/in-infusion, Q3Dx4).

On Fig shows the changes in body weight of nude mice bearing xenografts HCT-116 tumors carcinoma of the colon of a person, after treatment with 9,10-degidro-dEpoB (5 mg/kg, in/in-infusion, X3Dx4).

IG is a table with values IC 50for analogues epothilone in relation to cell growth CCRF-CEM.

On Fig shown metabolic stability analogues epothilone in vitro.

Fig is a table detailing therapeutic effects of various analogues epothilone against xenografts of human tumors in mice with 6-hour/in infusion.

On Fig shows the effect of 9,10-degidro-EpoB on the size of the tumors in nude mice bearing xenograft tumor HCT-116 carcinoma of the colon of a person (6 hours/in infusion, Q2Dx7).

On Fig shows the changes in body weight of nude mice bearing xenografts of tumor HCT-116 colon carcinoma intestine of man, after treatment with 9,10-degidro-EpoB and oxazol-EpoD (6-hour infusion, Q2Dx7).

On Fig shows the effect of 26-Cryptor-9,10-degidro-dEpoB and 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts A549/Taxol (6 hours/in infusion, Q2Dx4).

On Fig shows the effect of 9,10-degidro-dEpoB on the size of the tumors in nude mice bearing xenografts A549/Taxol (6 hours/in infusion, Q3Dx3).

On Fig shows the stability analogues epothilone in 20% plasma of mice/PBS.

On Fig shows the stability analogues epothilone 10% S9 fraction of liver person/PBS.

On Fig shows the chromatogram stability EpoD in 10% human liver S9/PBS.

On Fig presents tables that describe the effect of different an the log epothilone on the polymerization of microtubules in vitro at 37°C in the absence of GTP (A) and cytotoxicity of various analogues epothilone in the line of human lung cells A549 (B).

On Fig shown to stabilize the formation of microtubules epothilone at 35°C and 4°C.

On Fig shows therapeutic effect of 9,10-degidro-dEpoB nude mice bearing xenograft breast carcinoma human T (MX-1) (6-hour infusion, Q2Dx5).

On Fig shows the change in body weight of nude mice bearing xenograft breast carcinoma person (MX-1), after treatment with 9,10-degidro-dEpoB (6-hour infusion, Q2Dx8).

On Fig shows the change in body weight of nude mice bearing xenograft HCT-116, after treatment with 9,10-degidro-dEpoB (in/in-infusion, Q2Dx7).

On Fig shows therapeutic effect of 9,10-degidro-dEpoF, dEpoB and Taxol on the size of the tumors in nude mice bearing xenograft tumors of the breast carcinoma of the person (MX-1) (6 hours/in infusion, Q2Dx6).

On Fig shows the changes in body weight of nude mice bearing xenograft tumors of the breast carcinoma of the person (MX-1), after treatment with 9,10-degidro-dEpoF, dEpoB and Taxol (6-hour infusion, Q2Dx6).

On Fig shows therapeutic effect of 9,10-degidro-dEpoF and dEpoB nude mice bearing xenograft HCT-116 colon carcinoma intestine of man (6-hour infusion, Q2Dx8).

On Fig shows the changes in body weight of nude mice bearing xenograft HCT-116, after treatment with 9,10-degidro-dEpoF and dEpoB (6-hour infusion, Q2Dx8).

On Fig shown therapeutic deistvie,10-dihydro-dEpoF and dEpoB in mice nude, bearing xenograft resistant to Taxol of human lung carcinoma (A549/Taxol) (6-hour infusion, Q2Dx5).

On Fig shows the changes in body weight of nude mice bearing xenograft resistant to Taxol of human lung carcinoma (A549/Taxol), after treatment with 9,10-degidro-dEpoF and dEpoB (6-hour infusion, Q2Dx5).

Fig is a table comparing the effectiveness of various analogues epothilone in relation to inhibition of tumor growth in vitro and the relative therapeutic index.

On Fig shows therapeutic effect of 9,10-degidro-dEpoB nude mice bearing xenograft MX-1 (Q3Dx9, 6-hour/in infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft MX-1, after treatment with 9,10-degidro-dEpoB (Q3Dx9, 6-hour/in infusion).

On Fig shows therapeutic effect of 9,10-dehydroemetine B in nude mice bearing xenograft MX-1 (Q3Dx9, 6-hour infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft MX-1, after treatment with 9,10-dehydroemetine B (Q3Dx9, 6-hour/in infusion).

On Fig shown therapeutic effects of low doses of 26 trifter-9,10-degidro-dEpoB nude mice bearing xenograft MX-1 (6 hours/in infusion, Q2Dx12).

On Fig shows the changes in body weight of nude mice bearing xenograft MX-1, after treatment with low doses of 26-trip the EOS-9,10-degidro-dEpoB (6 hours/in infusion, Q2Dx12).

On Fig shown chemotherapeutic action of analogues epothilone against xenografts of human tumors in nude mice. The tumor tissue (40-50 mg) were implanted p/day 0. Treatment was started when the tumor size reached approximately 100 mm3or more, as specified. All processing, which is indicated by arrows, was performed using 6-hour/in infusion via the tail vein, using minicaster and programmable pump, as described previously (Su, D.-S., et al, Angew. Chem. Int. Ed. 1997, 36, 2093; Chou, T. C. et al. Proc. Natl. Acad. Sci. USA. 1998, 95, 15798; each publication included in this description by reference). Each dosing group consisted of four or more mice. Body weight was called total body mass minus the mass of the tumor, suggesting that 1 mm3the tumor is equal to 1 mg of tumor tissue. A. Xenograft breast carcinoma MX-1, treated with low doses of 25 trifter-(E)-9,10-degidro-12,13-disoxaril (10 mg/kg) when compared with the treatments in table 1 (20 mg/kg and 30 mg/kg). B. Large xenografts MX-1 (500 mm3) was treated with 25 trifter-(E)-9,10-degidro-12,13-disoxaril (25 mg/kg) and dEpoB 30 mg/kg). C. Slowly growing xenograft lung carcinoma A549 treated with 25 trifter-(E)-9,10-degidro-12,13-disoxaril (25 mg/kg) and dEpoB 30 mg/kg). D. Xenograft A549/Taxol (44-fold resistance to paclitaxel in vitro)treated 25 trifter-(E)-9,10-degidro-12,13-is ezekieru (20 mg/kg) and (E)-9,10-degidro-12,13-disoxaril (4 mg/kg). Processing deH-dEpoB on the 28th day was skipped due to a noticeable and rapid weight loss.

On Fig depicts the synthesis of C-21 modified 9,10-(E)-dehydroemetine. On figa shows the synthesis of 26-Cryptor-21-methylamino-9,10-(E)-degidro-12,13-desoxyepothilone B. FIGU is a scheme of synthesis to obtain 26 trifter-21-amino-9,10-(E)-degidro-12,13-desoxyepothilone B as an intermediate product in the synthesis of 26-Cryptor-21-dimethylamino-9,10-(E)-degidro-12,13-desoxyepothilone B.

Fig is a table with values IC50C-21 modified epothilone in relation to the line of tumor cells CCRF-CEM and its Pollini multidrug-resistant.

On Fig shown a therapeutic effect on 26 trifter-9,10-degidro-dEpoB and Taxol in nude mice bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM (6 hours/in infusion, Q2Dx8).

On Fig shows the changes in body weight of nude mice bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM, after treatment 25 trifter-9,10-degidro-dEpoB and Taxol (6 hours/in infusion, Q2Dx8).

On Fig shown a therapeutic effect on 26 trifter-9, 10-dehydro-dEpoB and Taxol in nude mice bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM/Taxol (resistant to Taxol) (6 hours/in infusion, Q2Dx7, x5).

On Fig shown ISM is in body weight of nude mice, bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM/Taxol (resistant to Taxol), after treatment 26 trifter-9,10-degidro-dEpoB and Taxol (6 hours/in infusion, Q2Dx7, x5).

On Fig shown a therapeutic effect on 26 trifter-9,10-degidro-dEpoB and Taxol in nude mice bearing xenograft carcinoma of the colon of human HCT-116 (Q2Dx4, x2, 6-hour/in infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft carcinoma of the colon of human HCT-116, after treatment 26 trifter-9,10-degidro-dEpoB and Taxol (Q2Dx4, x2, 6-hour/in infusion).

On Fig shows therapeutic effect of 9,10-degidro-EpoB nude mice bearing xenograft MX-1 (6 hours/in infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft breast carcinoma human MX-1, after treatment with 9,10-degidro-EpoB (6-hour infusion in/in infusion).

On Fig shows therapeutic effect of 9,10-degidro-EpoB nude mice bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM/Taxol (resistant to Taxol) (6 hours/in infusion, Q3Dx5, x2).

On Fig shows the changes in body weight of nude mice bearing xenograft T-cell lymphoblastic human leukemia CCRF-CEM/Taxol (resistant to Taxol), after treatment with 9,10-degidro-EpoB (6 hours/in infusion, Q3Dx5, x2).

On Phi is .72 shown a therapeutic effect on 26 trifter-dEpoB and 26 trifter-9,10-degidro-dEpoF nude mice, bearing xenograft breast carcinoma human MX-1 (Q2Dx11/injection).

On Fig shows the changes in body weight of nude mice bearing xenograft breast carcinoma human MX-1, after treatment 26 trifter-dEpoB and 26 trifter-9,10-degidro-dEpoF (Q2Dx11/injection).

On Fig shows therapeutic effect of 9,10-degidro-dEpoB nude mice bearing xenograft breast carcinoma human MX-1 (Q3Dx9, 6-hour/in infusion).

On Fig shows the changes in body weight of nude mice bearing xenograft breast carcinoma human MX-1, after treatment with 9,10-degidro-dEpoB (Q3Dx9, 6-hour/in infusion).

On Fig shown a therapeutic effect on 26 trifter-9,10-degidro-dEpoF nude mice bearing xenograft of human lung carcinoma (MX-1) (6 hours/in infusion and/injection).

On Fig shows the changes in body weight of nude mice bearing xenograft MX-1, after treatment 26 trifter-9,10-degidro-dEpoF (6 hours/in infusion and/injection).

Definitions

Some proposed in the present invention compounds and determine the specific functional groups are also described in more detail below. In this invention, the chemical elements are identified in accordance with the Periodic table of the elements, CAS version, Handbook of Chemistry and Physics, 75thEd., inside cover, and concr the local functional group, as a rule, determine, as described in this application. In addition, General principles of organic chemistry, as well as specific functional residues and chemical activity are described in "Organic Chemistry", Thomas Sorrel, University Science Books, Sausalito: 1999, the full contents of which are incorporated in this description by reference. In addition, the person skilled in the art will understand that the methods of synthesis which are described in this description, a number of security groups. The term "protective group"as used herein, is meant that a particular functional residue, for example O, S or N, is temporarily blocked so that a reaction was carried out selectively at another reactive site in a multifunctional compound. In preferred embodiments, the protective group selectively communicates with a good solution giving a protected substrate that is stable to the projected reactions; the protective group must be selectively removed in good yield readily available, preferably nontoxic reagents that do not affect other functional groups; protective group forms an easily separable derivative (more preferably without the formation of new stereogenic centers); and the protective group has a minimum of additional functionality to avoid further reactionists. As detailed in this description, you can use protective group oxygen, sulphur, nitrogen and carbon. Examples of protective groups described in detail in this description, however, it will be clear that this does not mean that the invention is limited to these protective groups; or a variety of additional equivalent protective groups can be easily identified using the above criteria, and use the method proposed in this invention. In addition, a number of protective groups described in "Protective Groups in Organic Synthesis", Third Ed. Greene, T. W. and Wuts, P.G., Eds., John Wiley and Sons, New York: 1999, the full contents of which are hereby incorporated by reference.

It will be understood that the compounds listed in the description, can be replaced with any number of substituents or functional residues. In General, the term "substituted", preceded or not preceded by the term "optional", and the deputies, included in the formula according to this invention, refers to the replacement of hydrogen radicals in a given structure with the radical of a particular Deputy. In the case when in any given structure may be substituted for more than one position more than one Deputy, selected from a particular group, the substituents in all positions can be either equal or different. Used in this described and the meaning it is supposed, the term "substituted" includes all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. In the present invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described in this publication, which satisfy the valences of the heteroatoms. In addition, it is understood that the invention is in no way limited by the permissible substituents of organic compounds. Combinations of substituents and variables proposed in this invention, preferable are combinations that lead to the formation of stable compounds applicable for the treatment of, for example, proliferative disorders, including, but not limited specified, malignant tumor. The term "stable", as used herein, preferably refers to compounds which possess stability sufficient to allow manufacture and which maintain the integrity of the connection during a period of time sufficient for registration, and preferably in the course the second time period, sufficient for his purposes, specified in detail in this specification.

The term "aliphatic" used in this sense, includes both saturated and unsaturated, straight chain (i.e. unbranched), branched, cyclic, or polycyclic aliphatic hydrocarbons, which are optionally substituted by one or more functional groups. As will be clear to the person skilled in the art, this description assumes that the term "aliphatic" includes, but is not limited to specified, alkyl, alkanniny, alkynylaryl, cycloalkenyl, cycloalkenyl and cycloalkyl residues. Thus, as used in this description, the term "alkyl" includes an unbranched, branched and cyclic alkyl groups. A similar condition applies to other General terms such as "alkenyl", "quinil" and the like. In addition, as used in this description, the terms "alkyl", "alkenyl", "quinil" and the like encompass both substituted and unsubstituted groups. In some embodiments, used in this description, the term "lower alkyl" is used to indicate alkyl groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched)having 1-6 carbon atoms.

In some embodiments, the alkyl, and keyline and alkyline group, used in the invention contain 1-20 aliphatic carbon atoms. In some other embodiments, the alkyl, alkeline and alkyline group used in the invention contain 1-10 aliphatic carbon atoms. In other embodiments, the alkyl, alkeline and alkyline group used in the invention contain 1-8 aliphatic carbon atoms. In the following embodiments, alkyl, alkeline and alkyline group used in the invention contain 1-6 aliphatic carbon atoms. In addition, in other embodiments, the alkyl, alkeline and alkyline group used in the invention contain 1-4 carbon atoms. Thus, the illustrative aliphatic groups include, but are not limited, for example, residues of methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -CH2-cyclopropyl, allyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclobutyl, -CH2-cyclobutyl, n-pentile, second-pentile, isopentyl, tert-pentile, cyclopentyl, -CH2-cyclopentyl, n-hexyl, sec-hexyl, cyclohexyl, -CH2-cyclohexyl and the like, which may also bear one or more substituents. Alkeneamine groups include, but are not limited, for example, ethynyl, propenyl, butenyl, 1-methyl-2-butene-1-yl and the like. Typical alkyline groups include, but are not limited to the shown, ethinyl, 2-PROPYNYL (propargyl), 1-PROPYNYL and the like.

The term "alkoxygroup" or "thioalkyl", as used herein, refers to an alkyl group as defined previously associated with the remainder of the parent molecule through an oxygen atom or through a sulfur atom. In some embodiments, the alkyl group contains 1-20 aliphatic carbon atoms. In some other embodiments, the alkyl group contains 1-10 aliphatic carbon atoms. In other embodiments, the alkyl, alkeline and alkyline group used in the invention contain 1-8 aliphatic carbon atoms. In the following embodiments, the alkyl group contains 1-6 aliphatic carbon atoms. In other embodiments, the alkyl group contains 1-4 aliphatic carbon atoms. Examples of alkoxygroup include, but are not limited to, methoxy, ethoxy-, propoxy-, isopropoxy, h-butoxy-, tert-butoxy, neopentane - and n-hexachrome. Examples of thioalkyl include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, h-butylthiourea and the like.

The term "alkylamino" refers to a group having the structure-other', where R' is alkyl, which is defined in this description. In some embodiments, the alkyl group contains 1-20 aliphatic carbon atoms. In some other embodiments, an alkyl group content is t 1-10 aliphatic carbon atoms. In other embodiments, the alkyl, alkeline and alkyline group used in the invention contain 1-8 aliphatic carbon atoms. In the following embodiments, the alkyl group contains 1-6 aliphatic carbon atoms. In other embodiments, the alkyl group contains 1-4 aliphatic carbon atoms. Examples of alkylamino include, but are not limited to, methylamino, ethylamino-from-propylamino and the like.

Some examples of substituents of the above described aliphatic (and other) residues of the compounds proposed in the invention include, but are not limited to, aliphatic group; heteroaromatics group; aryl; heteroaryl; arylalkyl; heteroaromatic; alkoxygroup; alloctype; heteroalkyl; heteroepitaxy; alkylthiols; killigrew; heterolytic; heterogroup; F; Cl; Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHCl2; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(RX); -CON(RX)2; -OC(O)RX; -OCO2RX; -OCON(RX)2; -N(RX)2; -S(O)2RX; -NRX(CO)RXwhere in each case RXindependently includes, but is not limited to specified, aliphatic group, heteroaromatics group, aryl, heteroaryl, arylalkyl or heteroaryl the sludge, where any of the aliphatic, heteroalicyclic, arylalkyl or heteroarylboronic substituents described above and in this case, can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and in this case, can be substituted or unsubstituted. Additional examples of generally applicable substituents are illustrated by the concrete variants shown in the examples provided in this description.

The terms "aryl" and "heteroaryl"used in this description refer to stable mono - or polycyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated residues having preferably 3-14 carbon atoms, each of which may be substituted or unsubstituted. The substituents include, without limitation, any of the previously mentioned substituents, i.e. substituents listed for aliphatic residues or other residues that are described in this publication, which lead to the formation of stable compounds. In some embodiments of this invention, "aryl" refers to mono - or bicyclic carbocyclic ring system having one or two aromatic rings including, but not limited to decrees of the data, phenyl, naphthyl, tetrahydronaphthyl, indolyl, indanyl and the like. In some embodiments of this invention, the term "heteroaryl", as used herein, refers to a cyclic aromatic radical having from five to ten atoms in the cycle, of which one atom in the cycle selected from S, O and N; zero, one, or two atoms in the cycle are additional heteroatoms independently selected from S, O and N; and the remaining atoms in the cycle are carbon atoms, and the radical is linked to the rest of the molecule through any atom in the cycle, such as pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, chinoline, ethenolysis and the like.

It will be clear that aryl and heteroaryl groups (including bicyclic aryl groups) can be unsubstituted or substituted, the substitution includes replacement of them in one, two or three of the hydrogen atoms is independently any one or more of the following residues, including without limitation: aliphatic group; heteroaromatics group; aryl; heteroaryl; arylalkyl; heteroaromatic; alkoxygroup; alloctype; heteroalkyl; heteroepitaxy; alkylthiols; killigrew; heterolytic; heteroaryl is tigroup; F; Cl; Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHCl2; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(RX); -CON(RX)2; -OC(O)RX; -OCO2RX; -OCON(RX)2; -N(RX)2; -S(O)2RX; -NRX(CO)RXwhere in each case RXindependently includes, but is not limited to specified, aliphatic group, heteroaromatics group, aryl, heteroaryl, arylalkyl or heteroaromatic, where any of the aliphatic, heteroalicyclic, arylalkyl or heteroarylboronic substituents described above and in this case, can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and in this case, can be substituted or unsubstituted. Additional examples of generally applicable substituents are illustrated by the concrete variants shown in the examples provided in this description.

The term "cycloalkyl"used in this description, in particular, refers to groups having from three to seven, preferably from three to ten carbon atoms. Suitable cycloalkyl include, but are not limited, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl itomo such, which, as in the case of other aliphatic, heteroalicyclic or heterocyclic residue may be optionally substituted by substituents, including without limitation aliphatic group; heteroaromatics group; aryl; heteroaryl; arylalkyl; heteroaromatic; alkoxygroup; alloctype; heteroalkyl; heteroepitaxy; alkylthiols; killigrew; heterolytic; heterogroup; F; Cl; Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHCl2; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(RX); -CON(RX)2; -OC(O)RX; -OCO2RX; -OCON(RX)2; -N(RX)2; -S(O)2RX; -NRX(CO)RXwhere in each case RXindependently includes, but is not limited to specified, aliphatic group, heteroaromatics group, aryl, heteroaryl, arylalkyl or heteroaromatic, where any of the aliphatic, heteroalicyclic, arylalkyl or heteroarylboronic substituents described above and in this case, can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and in this case, can be substituted or unsubstituted. Additional examples of Oba is but applicable substituents are illustrated by the concrete variants shown in the examples provided in this description.

The term "heteroaromatics", as used herein, refers to aliphatic residues, which contain one or more atoms of oxygen, sulfur, nitrogen, phosphorus or silicon, for example, instead of carbon atoms. Heteroaromatics residues can be branched, unbranched, cyclic or acyclic, and include saturated and unsaturated heterocycles, such as morpholinopropan, pyrrolidinyl, etc. In some embodiments heteroaromatics residues substituted by independent replacement in one or more hydrogen atoms of one or more residues, including, without limitation, aliphatic group; heteroaromatics group; aryl; heteroaryl; arylalkyl; heteroaromatic; alkoxygroup; alloctype; heteroalkyl; heteroepitaxy; alkylthiols; killigrew; heterolytic; heterogroup; F; Cl; Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHCl2; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(RX); -CON(RX)2; -OC(O)RX; -OCO2RX; -OCON(RX)2; -N(RX)2; -S(O)2RX; -NRX(CO)RXwhere in each case RXindependently includes, but is not limited to specified, and efficency group, heteroaromatics group, aryl, heteroaryl, arylalkyl or heteroaromatic, where any of the aliphatic, heteroalicyclic, arylalkyl or heteroarylboronic substituents described above and in this case, can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and in this case, can be substituted or unsubstituted. Additional examples of generally applicable substituents are illustrated by the concrete variants shown in the examples provided in this description.

The term "halogen", as used herein, refers to an atom selected from fluorine, chlorine, bromine and iodine.

The term "halogenated" means an alkyl group as defined above having one, two or three related halogen atoms, examples of which are groups such as chloromethyl, bromacil, trifluoromethyl and the like.

The term "heteroseksualci" or "heterocycle", as used herein, refers to non-aromatic 5-, 6 - or 7-membered cycle or polycyclic group, including, without limitation, bi - or tricyclic group containing a condensed six-membered ring having one to three heteroatoms independently selected from oxygen, sulfur and the PTA, where (i) each 5-membered cycle is from 0 to 1 double bonds and each 6-membered cycle is from 0 to 2 double bonds, (ii) the heteroatoms nitrogen and sulfur may not necessarily be oxidized, (iii) the nitrogen heteroatom may not necessarily Quaternary and (iv) any of the above heterocyclic rings may be fused with benzene ring. Typical heterocycles include, but are not limited, pyrrolidinyl, pyrazolyl, pyrazolidine, imidazoline, imidazolidine, piperidine, piperazinil, oxazolidinyl, isoxazolidine, morpholine, thiazolidine, isothiazolinones and tetrahydrofuryl. In some embodiments, uses the term "substituted heterocytolysine or heterocyclic group" used in this description, the term refers to geteroseksualnoe or heterocyclic group as defined above, with the substitution therein of one, two, or three hydrogen atoms without limitation aliphatic group; heteroaromatics group; an aryl; heteroaryl; arylalkyl; heteroallyl; alkoxygroup; arroceros; heteralsexual; heteroepitaxial; alkylthiol; arylthioureas; heteroalicyclic; heteroanalogues; F; Cl; Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHCl2; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)Rx ; -CO2(Rx); -CON(Rx)2; -OC(O)Rx; -OCO2Rx; -OCON(Rx)2; -N(Rx)2; -S(O)2Rx; -NRx(CO)Rxwhere in each case Rxindependently includes, but is not limited to specified, aliphatic group, heteroaromatics group, aryl, heteroaryl, arylalkyl or heteroaromatic, where any of the aliphatic, heteroalicyclic, arylalkyl or heteroarylboronic substituents described above and in this case, can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and in this case, can be substituted or unsubstituted. Additional examples of commonly used substituents illustrated with specific options shown in the examples provided in this description.

The term "labeled"as used herein, means that the compound has at least one associated element, isotope or chemical compound that makes it possible to check the connection. In General, the label can be attributed to three classes: a) isotopic labels, which may be radioactive or heavy isotopes, including, but not limited to,the2H,3H,32P,35S67Ga99mTc (Tc-99m),111In123 125I169Yb186Re; b) immune labels, which may be antibodies or antigens; and c) colored or fluorescent dyes. It will be clear that the label can be included in the connection in any position which does not prevent the biological activity or characteristic of a compound that register. In some embodiments of the invention use photoaffinity tagging for the direct determination of intermolecular interactions in biological systems (for example, to investigate the binding site epothilone in the tubulin dimer). You can use many fotoforum, most of which are based on photopreteen of diazo compounds, azides or diazirine in nitrene or carbenes (see Bayley, H., Photogenerated Reagents in Biochemistry and Molecular Biology (1983), Elsevier, Amsterdam.), the full contents of this publication are included in this description by reference. In some embodiments of the invention used photoaffinity labels are ortho-, meta - and para-azidobenzoyl substituted by one or more residues of halogen, including, but not limited to, 4-azido-2,3,5,6-tetrafluorobenzoic acid.

"Polymer": the Term "polymer", as used herein, refers to compositions containing chain which may be open, closed, linear, branched or crosslinked repeated e is the INIC (monomers), which may be the same or different. It will be understood that in some embodiments, the term "polymer" refers to a biopolymer, which, as implied, are natural polymeric materials, or based on materials found in nature, including, but not limited to, nucleic acids, peptides and their mimetics. In some other embodiments, the term "polymer" refers to synthetic polymers, such as biodegradable polymers or other polymer materials. It will be understood that the polymeric solid carriers are also included in the polymers proposed in this invention. Proposed in the invention compounds can be associated with a polymeric carriers and, therefore, some synthetic modifications can be carried out on solid phase. Used in this description, the term "solid support" includes, but is not limited to specified, pellets, disks, capillaries, hollow fibers, needles, pins, solid fibers, cellulose beads, porous glass beads, silica gels, polystyrene beads optionally crosslinked with divinylbenzene beads of grafted copolymers, polyacrylamide beads, latex beads, dimethylacrylamide balls, optional cross-stitched with N, N'-bis-acryloylmorpholine, and glass particles coated with hydrophobic polymer. Special features : the students in this area will be clear, the choice of a specific solid media will be limited media compatibility with the chemical reactions. Examples of solid carrier is the amino Tentagel, composition 1) polystyrene balls, cross-crosslinked with divinylbenzene, and (2) PEG (polyethylene glycol). Tentagel is particularly applicable to solid media because it provides universal support for use in the analysis on balls and no balls, and a well-swells in solvents ranging from toluene to water.

The description of some embodiments of the invention

Recognizing the need to develop new and effective methods of treatment of malignant tumors, the present invention offers a new method of synthesis that allows access to the macrocycle, with a wide range of biological and pharmacological activity, as well as new compounds with such activity, new therapeutic compositions and methods of using such compounds and compositions.

In some embodiments proposed in the invention compounds applicable in the treatment of malignant tumors. We offer some of the invention compounds have a cytotoxic or growth inhibitory effect on cell lines of malignant tumors, demonstrate ability to polimerizuet tubulin and reliable in order to lyse the structure of microtubules and/or causing the reduction or disappearance of tumors in xenografts models of malignant tumor cells. In some embodiments, compounds can be reduced or minimal side effects, including toxicity to vital organs, nausea, vomiting, diarrhea, alopecia, weight loss, weight increase toxicity to the liver, skin disorders, etc. Connections are also easier can be prepared in the form of compositions due to the high water solubility, reduced toxicity, increased therapeutic range, high efficiency, etc.

General description offered in the invention compounds

Compounds of the present invention include compounds of General formula (0) and (0'), which are further defined below:

in which R0means substituted or unsubstituted aryl, heteroaryl, arylalkyl, killkenny, arylalkylamines, heteroarylboronic, heteroarylboronic or heteroarylboronic balance; in some embodiments, R0means killkenny, killkenny, heteroarylboronic or heteroarylboronic balance; in other embodiments, R0means heteroarylboronic balance; in some embodiments, R0means heteroarylboronic balance; in other embodiments, R0means a 5-7 membered aryl or heteroaryl residue; in other embodiments, R0means 8-12-member of the St bicyclic aryl or heteroaryl residue; in the following embodiments, R0means bicyclic residue, in which the phenyl cycle condensed with a heteroaryl or aryl residue; in other embodiments, R0means bicyclic residue, in which the phenyl ring condensed with the residue of thiazole, oxazole or imidazole; in other embodiments, R0means substituted or unsubstituted phenyl residue;

R3and R4each independently denote hydrogen; or substituted or unsubstituted, linear or branched, cyclic or acyclic aliphatic, heteroaromatics radical, aryl, heteroaryl, arylalkyl or heteroarylboronic residue, optionally substituted by one or more groups of hydroxyl, protected hydroxyl, alkoxygroup, carboxypropyl, carboxaldehyde, linear or branched alkyl or cyclic acetal, fluorine, amino, protected amino, amino substituted by one or two alkyl or aryl residues, N-hydroxyisopropyl or N-alkoxyimino; in some embodiments, R3and R4each independently denote a hydrogen, fluorine or lower alkyl; in other embodiments, R3and R4each independently mean hydrogen or methyl; in other embodiments, R3means methyl, and R4means hydrogen;

R5and R6each independently mean hydrogen or a protective group; in some embodiments, R5and R6both signify hydrogen;

X represents O, S, C(R7)2or NR7where in each case R7independently means hydrogen or lower alkyl; in some embodiments, X is O; in other embodiments, X is NH;

Y represents O, S, NH, C(R7)2CH2N(R7or NH, where in each case independently of R7means hydrogen or lower alkyl; in some embodiments, Y represents O; in other embodiments, Y represents NH; in other embodiments, Y represents CH2;

each R8independently means hydrogen, halogen, hydroxyl, alkoxygroup, amino group, dialkylamino, alkylamino, fluorine, cyano or substituted or unsubstituted, linear or branched, cyclic or acyclic aliphatic, heteroaromatics radical, aryl, heteroaryl, arylalkyl, killkenny, arylalkylamine or heteroarylboronic, heteroarylboronic, heteroarylboronic residue, optionally substituted by one or more substituents of hydroxyl, protected hydroxyl, alkoxygroup, carboxypropyl, carboxaldehyde, linear or branched alkyl or cyclic acetal, fluorine, amino, protected amino, amino substituted by one or two alkyl or aryl residues, N-hydroxyisopropyl and the and N-alkoxyimino; in some embodiments, R8means hydrogen; in other embodiments, R8means hydroxyl; in other embodiments, R8means fluorine, in the following embodiments, R8means lower alkyl, such as methyl; in other embodiments, R8means-CF3, -CF2H or CFH2; in other embodiments, R8means perfluorinated or fluorinated alkyl group; in other embodiments, R8means halogenated or perhalogenated alkyl group;

R9and R10each independently denote hydrogen; or substituted or unsubstituted, linear or branched, cyclic or acyclic aliphatic, heteroaromatics radical, aryl, heteroaryl, aryl, arylalkyl, killkenny, arylalkylamines, heteroarylboronic, heteroarylboronic or heteroarylboronic residue, optionally substituted by one or more substituents of hydroxyl, protected hydroxyl, alkoxygroup, carboxypropyl, carboxaldehyde, linear or branched alkyl or cyclic acetal, fluorine, amino, protected amino, amino substituted by one or two alkyl or aryl residues, N-hydroxyisopropyl or N-alkoxyimino; in some embodiments, one of R9and R10means methyl; in other embodiments, the BA R 9and R10means methyl; in other embodiments, one of R9and R10means methyl, and the other denotes hydrogen; in other embodiments, both R9and R10mean hydrogen;

RBindependently for each case denotes hydrogen; halogen; -ORB'; -SRB'; -N(RB')2; -C(O)ORB'; -C(O)RB'; -CONHRB'; -O(C=O)RB'; -O(C=O)ORB'; -NRB'(C=O)RB'; N3; N2RB'; cyclic acetal; or cyclic or acyclic, linear or branched aliphatic, heteroaromatics radical, aryl or heteroaryl, optionally substituted by one or more groups from hydrogen; halogen; -ORB'; -SRB'; -N(RB')2; -C(O)ORB'; -C(O)RB'; -CONHRB'; -O(C=O)RB'; -O(C=O)ORB'; -NRB'(C=O)RB'; N3; N2RB'; cyclic acetal; or cyclic or acyclic, linear or branched substituted or unsubstituted aliphatic, heteroaromatics radical, aryl or heteroaryl residue; or is epothilones, desoxyepothilone, or their equivalents; or is a polymer; carbohydrate; photoaffinity label; or radioactive label; in some embodiments, RBmeans hydrogen,, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or qi is logical, each unsubstituted or optionally substituted occurring one or more times by groups of halogen, -OH, -ORB', NH2or N(RB')2or combinations thereof, where in each case independently of RB'means hydrogen, alkyl, aryl, or a protective group, in other embodiments, RBmeans hydrogen, methyl or ethyl, in other embodiments, RBmeans methyl, in other embodiments,- CY3, -CHY2, -CH2Y, where Y means F, Br, Cl, I, ORB', OtherB'N(RB')2or SRB'; in other embodiments, RBmeans-CF3, -CH2F or CHF2; in other embodiments, RBmeans perfluorinated or fluorinated alkyl group; in other embodiments, RBmeans halogenated or perhalogenated alkyl group; in each case, RBindependently means hydrogen, a protective group; a linear or branched, substituted or unsubstituted, cyclic or acyclic, aliphatic, heteroaromatics radical, aryl, heteroaryl, arylalkyl, killkenny, arylalkylamines, heteroarylboronic, heteroarylboronic or heteroarylboronic balance;

m is 1, 2, 3 or 4, m is 1 or 2, in some embodiments, m is 1 in other embodiments;

and their pharmaceutically acceptable derivatives.

Connections proposed in the invention include the value of General formula (I) or (I'), which are further defined below:

in which R1means hydrogen or lower alkyl; in some embodiments, R1means methyl; in some embodiments, R1means-CF3, -CF2H or CH2F; in other embodiments, R1means perfluorinated or fluorinated alkyl group; in other embodiments, R1means halogenated or perhalogenated alkyl group;

R2means substituted or unsubstituted aryl, heteroaryl, arylalkyl or heteroarylboronic balance; in some embodiments, R2means substituted or unsubstituted oxazole; in other embodiments, R2means substituted or an unsubstituted thiazole;

R3and R4each independently mean hydrogen; or substituted or unsubstituted, linear or branched, cyclic or acyclic aliphatic, heteroaromatics radical, aryl, heteroaryl, arylalkyl or heteroarylboronic residue, optionally substituted by one or more groups of hydroxyl, protected hydroxyl, alkoxygroup, carboxypropyl, carboxaldehyde, linear or branched alkyl or cyclic acetal, fluorine, amino, protected amino, amino substituted by one or two alkyl or ariline is, and remains, N-hydroxyisopropyl or N-alkoxyimino; in some embodiments, R3and R4each independently denote a hydrogen, fluorine or lower alkyl; in other embodiments, R3and R4each independently mean hydrogen or methyl; in the following embodiments, R3means methyl, and R4means hydrogen;

R5and R6each independently mean hydrogen or a protective group; in some embodiments, R5and R6both signify hydrogen;

X represents O, S, C(R7)2or NR7where in each case R7independently means hydrogen or lower alkyl; in some embodiments, X is O; in other embodiments, X is NH;

RBindependently for each case denotes hydrogen; halogen; -ORB'; -SRB'; -N(RB')2; -C(O)ORB'; -C(O)RB'; -CONHRB'; -O(C=O)RB'; -O(C=O)ORB'; -NRB'(C=O)RB'; N3; N2RB'; cyclic acetal; or cyclic or acyclic, linear or branched aliphatic, heteroaromatics radial, aryl or heteroaryl, optionally substituted by one or more groups from hydrogen; halogen; -ORB'; -SRB'; -N(RB')2; -C(O)ORB'; -C(O)RB'; -CONHRB'; -O(C=O)RB'; -O(C=O)ORB'; -NRB'(C=O)RB'; N3; N2RB'; cyclic acetal; or cyclic or acyclic what about, linear or branched substituted or unsubstituted aliphatic, heteroaromatics radical, aryl or heteroaryl residue; or is epothilones, desoxyepothilone, or their equivalents; or is a polymer; carbohydrate; photoaffinity label; or radioactive label; in some embodiments, RBmeans hydrogen,, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, each of which is unsubstituted or optionally substituted occurring one or more times by groups of halogen, -OH, -ORB', NH2or N(RB')2or any combinations thereof, where in each case independently of RB'means hydrogen, alkyl, aryl, or a protective group, in other embodiments, RBmeans hydrogen, methyl or ethyl, in other embodiments, RBmeans methyl, in other embodiments, RBmeans-CF3, -CH2F or CHF2;

and their pharmaceutically acceptable derivatives.

In some embodiments, the compounds proposed in the invention include compounds of General formula (II) or (II')with the stereochemistry defined as follows:

where X, R1, R2, R3, R4, R5, R6, RBand X have the meanings defined above.

In some embodiments, X Osnach the em O. In other embodiments, X is NH. In other embodiments, X is CH2.

In some embodiments, R2means substituted or an unsubstituted thiazole. In some embodiments, R2means 2 methylthieno-4-yl. In other embodiments, R2means 2 hydroxymethylene-4-yl. In other embodiments, R2means 2 aminomethylation-4-yl. In other embodiments, R2means 2 tholeiltes-4-yl.

In some embodiments, R2means substituted or unsubstituted oxazole. In some embodiments, R2means 2 metaloxide-4-yl. In other embodiments, R2means 2 hydroxymethylamino-4-yl. In other embodiments, R2means 2 aminoethylamino-4-yl. In other embodiments, R2means 2 tilletiaceae-4-yl.

In some embodiments, RBmeans hydrogen, methyl, ethyl, -CF3, -CH2F, -CF2H. In some embodiments, RBmeans methyl. In other embodiments, RBmeans-CF3. In some embodiments, RBmeans hydrogen. In other embodiments, RBmeans ethyl.

Some preferred compounds include, for example:

Compounds proposed in this invention include compounds specifically listed above and described in this publication, and they are partially illustrated various classes, subgenera, and species disclosed elsewhere in this description.

The person skilled in the art it will be clear that in the compounds of the present invention may exist asymmetric centers. Thus, the proposed compounds and their pharmaceutical compositions can be in the form of an individual enantiomer, diastereoisomer or a geometric isomer, or may be in the form of mixtures of stereoisomers. In some embodiments proposed in the invention compounds are enantiomerically pure compounds. In some other embodiments, offers a mixture of stereoisomers or diastereoisomers.

It will be clear that some of the above classes and subclasses of compounds can exist in various isomeric forms. The invention includes compounds in the form of the CTD is lnyh isomers, essentially not containing other isomers, and alternative in the form of mixtures of various isomers, e.g., racemic mixtures of stereoisomers. In addition, the invention includes both (Z)-and (E)-isomers with respect to the double bond, unless otherwise stated. Thus, the proposed invention in connection, usually represented by the structures(0), (0'), (I), (I'), (II) and (II') include structures in which the double bond have a configuration (Z) or (E). In some preferred embodiments, the double bond at position C12-C13 is CIS - or Z-configuration. In some embodiments, the double bond at position C9-C10 is a TRANS - or E-configuration. In other embodiments, the double bond at position C12-C13 is CIS - or Z-configuration, and the double bond at position C9-C10 is a TRANS - or E-configuration. The invention also includes tautomers specific compounds described above.

In addition, this invention relates to a pharmaceutically acceptable derivative proposed in the invention compounds and methods of treatment of a subject with the use of these compounds, pharmaceutical compositions, or any of the specified in combination with one or more additional therapeutic agents. The term "pharmaceutically acceptable derivative", as used herein, means any farm is citiesi acceptable salt, ester or salt of ester of the claimed compounds or any other adduct or derivative which upon administration to the patient is able to give (directly or indirectly) a compound, which in all other respects described in this publication, or its metabolite or residue. Thus, the pharmaceutically acceptable derivatives, among other things, include prodrugs. A prodrug is a derivative of connection is usually reduced significantly pharmacological activity, which contains additional residue that is difficult to remove in vivo, giving the original molecule in the form of the pharmacologically active form. An example of a prodrug is an ester which is cleaved in vivo, giving the desired compound. Prodrugs of a number of compounds and substances and methods for derivatization of the original connection to create prodrugs, are known and can be adapted for use with this invention. Some examples of pharmaceutical compositions and pharmaceutically acceptable derivatives will be discussed in more detail in the description below.

Connections proposed in the invention that are of particular interest include compounds that:

- exert cytotoxic and growth inhibitory effects on cell lines of malignant tumors, supported n vitro, or in animal studies using acceptable scientific objectives xenograft models of malignant tumor cells;

- demonstrate ability to polimerizuet tubulin and stabilize structures, assembled from microtubules;

- demonstrate a minimum level of toxicity in relation to vital organs;

- lead to the disappearance of tumors in an acceptable scientific goal models xenografts tumor;

- lead to the reduction of tumors in an acceptable scientific goal models xenografts tumor;

- lead to the disappearance of tumors in an acceptable scientific goal models xenografts cells of malignant tumors and delayed relapse/or absence of tumor recurrence after stopping treatment;

provide temporary and reversible reduction of body weight and have a therapeutic effect in an acceptable scientific goal models xenografts tumor;

- show an increased solubility in water as compared with epothilone A, B, C or D or paclitaxel, or additionally, or alternatively have a solubility sufficient for cooking in the idea compositions in aqueous medium using a reduced share of cremophor; and/or

- demonstrate therapeu the practical profile (for example, optimum safety and therapeutic effect), which is better than the profile epothilone B, epothilone D or paclitaxel.

Many analogues epothilone that described above is obtained, characterized and tested as described in the examples in this description. Found that analogs of 9,10-dehydroemetine applicable for the treatment of malignant tumors, and, in particular, the compounds and found that they have one or more characteristic features listed above.

The technique of synthesis

Synthesis of some epothilones, desoxyepothilone and their analogues described previously (see U.S. patent 6242469, 6284781, 6300355, 6204388, 6316630 and 6369234; application for granting U.S. patents 09/797027, 09/796959 and 10/236135; and PCT publication no WO 99/01124, WO 99/43653, and WO 01/64650, the full contents of which are incorporated in this description by reference). Aware of the need for improved or additional methods of synthesis for effective education epothilones, desoxyepothilone and their counterparts in large quantities, the present invention offers an efficient and modular way of synthesis epothilones, desoxyepothilone and their analogues. Although the synthesis of some are given as examples of compounds described in the illustrative examples in this description, it will be clear that this method, in General, applicable to obtaining analogues and conjugates discussed is use, for each of the classes and subclasses described in this description.

In particular, the compounds 9,10-dehydroemetine proposed in this invention, can be obtained in a variety of ways using methods of synthesis which are applicable in the synthesis epothilones. In some embodiments, compounds obtained using convergent synthesis pathway. For example, epothilone it is possible to synthesize, receiving two or three intermediate product, which connect together, getting the desired connection. In one embodiment, one of the intermediate products is acyl part containing 1-9 carbon atoms, and the other intermediate product contains carbon atoms 10-15 and may also contain a side chain of thiazole. These two roughly equal parts epothilone can be joined together first using the esterification reaction between C-1 and the oxygen at C-15. Then the macrocycle can be closed using the reaction of formation of carbon-carbon, such as the Suzuki reaction mix or metathesis reaction with the closure cycle. In one embodiment, the final stage circuit cycle performed by using a metathesis reaction with the closure of the cycle with the formation of 9,10-double bond and the closure of the macrocycle. The metathesis reaction with snapping cycle performed by using metal-organic produce is RA, such as the catalyst of Grubbs, as shown in scheme 8 below. In some embodiments, 9,10-double bond restore or oxidize, or 9,10-double bond can be further functionalitywith to get additional derivatives epothilone.

In other embodiments, the final stage circuit cycle performed by using a metathesis reaction with the closure of the loop to form 12,13-double bond and to close the macrocycle. In some embodiments, 12,13-double bond restore or oxidize. In other embodiments, using the reaction of macroalbuminuria or macromechanical to form the macrocycle.

Some are given as examples of the reaction synthesis proposed in the invention compounds are presented in the figures and in the examples. As can be understood by the specialist, you can get many analogues and derivatives, using described in this publication, the methods of synthesis. For example, you can implement many stages of the synthesis with different protective groups or different substituents in the 16-membered cycle.

The pharmaceutical composition

This invention also relates to a pharmaceutical preparation containing at least one compound described above, or its pharmaceutically acceptable derivative, and these compounds are able to inhibit the growth or kill cells Slokas is only the tumor and some special interest options are able to inhibit the growth or kill cells of a malignant tumor multidrug resistance. In some embodiments, the pharmaceutical preparation also contains solubilizers or emulsifying agent, such as cremophor (polyoxyl 35 castor oil) or solutol (12-hydroxystearic of polyethylene glycol 660).

As discussed above, this invention relates to new compounds with anticancer and antiproliferative activity, and thus, the proposed invention in connection applicable for the treatment of malignant tumors. Accordingly, in another aspect, the present invention offers pharmaceutical compositions, these compositions contain any of the compounds described in this application, and optionally contain a pharmaceutically acceptable carrier. In some embodiments, these compositions optionally additionally contain one or more additional therapeutic agents. In some other embodiments, the additional therapeutic agent is an anti-cancer agent, which are discussed in detail in this specification.

Some compounds of this invention are suitable for treatment, can exist in free form or in the appropriate cases in the form of their pharmaceutically acceptable derivative. According to this invention pharmaceutically acceptable derivative includes, but is not limited to specified the output, pharmaceutically acceptable salts, esters, salts of such esters, or any other adduct or derivative which upon introduction to the needy in this the patient is able to give, directly or indirectly, a compound that in other respects described in this publication, or its metabolite or residue, for example, the prodrug.

Used in this description, the term "pharmaceutically acceptable salt" refers to salts that are based on informed medical opinion suitable for use in contact with human tissues and more disorganized animals without excessive toxicity, irritation, allergic response and the like and meet the reasonable value of benefit/risk. Pharmaceutically acceptable salts are well known in this field. For example, S. M. Berge, et al. describe pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 66: 1-19 (1977), included in this description by reference. Salts can be obtained in situ during the final isolation and purification of the compounds according to the invention or separately by the interaction of the functions of the free base with a suitable organic acid. Examples of pharmaceutically acceptable non-toxic acid additive salts are salts of an amino group formed with inorganic acids such as chlorestol the portly acid, Hydrobromic acid, phosphoric acid, sulfuric acid, Perlina acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in this field, such as ion exchange. Other pharmaceutically acceptable salts include salts, as adipate, alginate, ascorbate, aspartate, bansilalpet, benzoate, bisulfate, borate, butyrate, comfort, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulphate, aconsultant, formate, fumarate, glucoheptonate, glycerol, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonic, lactobionate, lactate, laurate, lauryl, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, para-toluensulfonate, undecanoate, valerate, and the like. Typical salts of alkaline or alkaline earth metals include sodium, lithium, potassium, calcium, magnesium and the like. These pharmaceutically acceptable salts include in an appropriate case, non-toxic cations ammonium, chetvertichnogo the ammonium and amine, formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkylsulfonate and arylsulfonate.

In addition, as used in this description, the term "pharmaceutically acceptable ester" refers to esters which are hydrolyzed in vivo and include esters, which easily disintegrate in the body, however, remains the original compound or its salt. Suitable ester groups include, for example, groups derived from pharmaceutically acceptable aliphatic carboxylic acids, especially alkanovykh, alkenovich, cycloalkanes acids and altanticist, in which alkyl or alkanniny the rest is mostly not more than 6 carbon atoms. The specific examples of esters include formate, acetate, propionate, butyrate, acrylates and ethylsuccinate.

In addition, the term "pharmaceutically acceptable prodrugs"as used herein, refers to those prodrugs of the compounds proposed in this invention, on the basis of informed medical opinion suitable for use in contact with human tissues and more disorganized animals without excessive toxicity, irritation, allergic response and the like, correspond to the reasonable value of benefit/risk and the effect is positive in the case of alleged use, and if possible, zwitterionic forms of the compounds proposed in the invention. The term "prodrug" refers to compounds that are rapidly transformed in vivo, giving the parent compound of the above formula, for example, by hydrolysis in blood. A comprehensive discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol.14 of the A. C. S. Symposium Series, and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both publications included in this description by reference.

As described above, the pharmaceutical compositions provided in this invention also include pharmaceutically acceptable carrier includes any and all solvents, diluents, or other liquid fillers, excipients for the formation of dispersions or suspensions, surface-active agents, agents, providing isotonicity solution, thickeners or emulsifiers, preservatives, solid binders, lubricants and the like, which are suitable for your desired dosage form. In Remington''s Pharmaceutical Sciences, Fifteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1975) describes the different media used for preparation of pharmaceutical compositions and known methods of obtaining them. Except, when any normal environment media mesowest is and antitumor compound according to the invention, for example, providing any undesirable biological effect or otherwise interacting with any other compound(s) of the pharmaceutical composition with an undesirable effect, its application is considered within the scope of this invention. Some examples of substances which can serve as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth gum; malt; gelatin; talc; cremophor; solutol; excipients such as cocoa butter, waxes for suppositories; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols, such as propylene glycol; esters, such as etiloleat and tillaart; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; ringer's solution; ethyl alcohol and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, and colouring agents, Genty, helps to release covering agents, sweeteners, corrigentov fragrances, preservatives and antioxidants can also be present in the composition in accordance with the decision of the expert preparing the composition.

Use of the compounds and pharmaceutical compositions

The invention also relates to a method of inhibiting tumor growth and/or metastasis of tumors. In some special interest embodiments, the invention relates to a method of treatment of malignant tumors by inhibiting tumor growth and/or metastasis of tumors in case of malignant tumor cells with multidrug resistance. The method consists of introducing a therapeutically effective amount of the compound or its pharmaceutically acceptable derivative needy in this subject (including, but not limited to, human or animal). In some embodiments, especially in the case of treatment of malignant tumors containing malignant cells with multidrug resistance, a therapeutically effective amount is an amount sufficient to kill or inhibit the growth of cell lines of malignant tumor multidrug resistance. In some embodiments, discov what's in the invention compounds applicable for the treatment of solid tumors.

Compounds and pharmaceutical compositions proposed in this invention can be used for the treatment and prevention of any disease or conditions, including proliferative diseases (e.g., cancer), autoimmune diseases (e.g. rheumatoid arthritis) and infection (e.g. bacterial, fungal etc). Compounds and pharmaceutical compositions can be entered to animals, preferably mammals (e.g., domesticated animals, cats, dogs, mice, rats), and more preferably humans. You can use any method of administration for delivery of the compounds of the pharmaceutical composition to an animal. In some embodiments, the compound or pharmaceutical composition is administered parenterally.

In another aspect, according to the treatment methods proposed in this invention, the tumor cell kill or inhibit their growth by contact of tumor cells with the proposed compound or composition, are described in this publication. Thus, in another aspect, the invention features a method of treating malignant tumors involving the introduction of a therapeutically effective amount proposed in the invention, the compounds or pharmaceutical compositions containing the proposed invention in connection, those in need who I am in this subject in such quantities and for such period of time, which is necessary to achieve the desired result. In some embodiments of this invention a "therapeutically effective amount" proposed in the invention, the compounds or pharmaceutical compositions represents a quantity that is effective for killing or inhibiting the growth of tumor cells. The compounds and compositions according to the method proposed in this invention can be administered using any amount and any route of administration effective for killing or inhibiting tumor cells. Thus, the expression "amount effective for killing or inhibiting the growth of tumor cells", as used herein, refers to the amount of the agent sufficient to kill or inhibit the growth of tumor cells. The exact quantity required will vary depending on the species, age and General condition of the subject, the severity of infection, specific antitumor agent, its mode of administration and the like. Antitumor compounds proposed in the invention, preferably prepared in a dosage form for ease of administration and consistency of the dose. The expression "dosage form"as used herein, refers to a physically discrete unit of protiva wholesale agent, suitable for the patient being treated. However, it will be clear that relative to the total daily use of compounds and compositions as proposed in the present invention, the solution will be to make an attending physician on the basis of informed medical opinion. The specific level of therapeutically effective dose for any particular patient or organism will depend on many factors, including the disease being treated and the severity of the disorder; activity of the specific compound; the specific composition; the age, body weight, General health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound; the duration of the treatment; drugs used in combination or simultaneously with the compound; and like factors well known in medicine.

In addition, after preparation of the composition with a suitable pharmaceutically acceptable carrier in a desired dosage, the pharmaceutical compositions proposed in the present invention, it is possible to enter to man and other animals orally, rectally, parenterally, intracisternally, intrawaginalno, intraperitoneally, locally (powders, ointments or drops), buccal, in the form of oral or called the high spray and the like, depending on the severity of the infection being treated. In some embodiments of the invention offer connections that are described in this publication, prepared by conjugation with water-soluble by chelators or water-soluble polymers such as polyethylene glycol as poly(1-glutamic acid) or poly(1-aspartic acid), which are described in U.S. patent 5977163, the full contents of which are included in this description by reference. In some embodiments proposed in the invention compounds can be administered orally or parenterally in a dose sufficient to deliver from about 0.001 mg/kg to 100 mg/kg, from about 0.01 mg/kg to 50 mg/kg, preferably from about 0.1 mg/kg to 40 mg/kg, preferably from about 0.5 mg/kg to 30 mg/kg, from about 0.01 mg/kg to 10 mg/kg, from about 0.1 mg/kg to 10 mg/kg and more preferably from about 1 mg/kg up to 25 mg/kg of body weight of the subject per day, once or several times per day to obtain the desired therapeutic effect. The required dose can be delivered every other day, every three days, every week, every two weeks, every three weeks or every four weeks. In some embodiments, the required dose can be delivered using multiple injections (e.g., two, three, four, five, six, seven, eight, nine or ten).

Liquid dose is rowanne forms for oral administration include, but not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in this field, such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn oil, oil of wheat germ, olive, castor and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and esters sorbitan and fatty acids and their mixtures. Besides inert diluents, the oral compositions can also contain adjuvants, such as moisturizers, emulsifiers and suspendresume agents, sweeteners, corrigentov and fragrances.

Injectable preparations, for example, sterile injectable aqueous or oily suspensions may be prepared according to known techniques using suitable dispersing and wetting agents and suspendida agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in netoc the ranks parenterally acceptable diluent or solvent, for example in the form of a solution in 1,3-butanediol. To an acceptable fillers and solvents that may be used include water, ringer's solution, U.S.P. and isotonic solution of sodium chloride. In addition, as a solvent or suspension medium is usually used sterile non-volatile oil. For this purpose you can use any soft fixed oils, including synthetic mono - or diglycerides. In addition, to obtain injectable use fatty acids such as oleic acid.

Injectable preparations can be sterilized, for example, by filtration through a filter that retains bacteria, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersing in sterile water or other sterile injectable medium prior to use.

To prolong the action of drugs, it is often desirable to slow the absorption of drugs delivered by subcutaneous or intramuscular injection. This can be done using a liquid suspension of crystalline or amorphous material with poor water solubility. The degree of absorption of the drug in this case depends on the degree of dissolution, which in turn may depend on the size of the crystals and the crystal is a political form. Alternative slow the absorption of parenteral introduced forms of drugs is achieved by dissolving or suspendirovanie medicines in the oil filler. Injectable form a delayed suction prepared by education matrices for microencapsulation drug in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer used, the rate of release of drug can be adjusted. Examples of other biodegradable polymers include complex poly(orthoevra) and poly(anhydrides). Injectable drugs slow suction also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal injection preferably are suppositories which can be prepared by mixing the compounds proposed in this invention with suitable non-irritating with excipients or carriers such as cocoa butter, polyethylene glycol or wax for suppositories, which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginalnogo and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms the active compound is mixed, with at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or dry diluents, such as starches, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and the Arabian gum, (c) wetting means, such as glycerol, d) dezinfeciruyuhimi agents such as agar-agar, calcium carbonate, potato starch or starch from tapioca, alginic acid, certain silicates and sodium carbonate, e) slow dissolving agents, such as paraffin, f) accelerators suction, such as Quaternary ammonium compounds, g) humectants, such as, for example, cetyl alcohol and glycerylmonostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricating agents such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills dosage form may also contain buffering agents.

the solid compositions of a similar type may also be employed as fillers in soft and hard gelatin capsules using such excipients, as lactose or milk sugar, as well as glycols of high molecular weight and the like. Solid dosage forms of tablets, pills, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the field of preparation of pharmaceutical compositions. They may not necessarily contain contrast agents and can also be of such composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract does not necessarily slow. Examples of encapsulating compositions, which can be used include polymeric substances and waxes. Solid compositions of a similar type can also be used as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar, as well as glycols of high molecular weight and the like.

The active compounds can also be microencapsulating form with one or more excipients described above. Solid dosage forms of tablets, pills, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, coatings for controlled release, etc is affected coating, well known in the field of preparation of pharmaceutical compositions. In such solid dosage forms the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain, and it is normal practice, additional substances other than inert diluents, for example, the lubricant for tableting and other AIDS for tableting, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills dosage forms may also contain buffering agents. They may not necessarily contain contrast agents and can also be of such composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract does not necessarily slow. Examples of encapsulating compositions, which can be used include polymeric substances and waxes.

Dosage forms for local or transdermal injection of the compounds proposed in this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers or patches. The active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed to the sideboards or buffers, which may be required. Ophthalmic drug, ear drops and eye drops are also included in the scope of this invention. In addition, this invention covers the use of transdermal patches, which have the added advantage of providing controlled delivery of compounds into the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. You can also use the amps suction to increase the flow connection through the skin. Speed can be controlled either by providing speed control membrane, or dispersive compound in a polymer matrix or gel.

As discussed above, the compounds proposed in this invention, applicable as antitumor agents, and thus may be applicable in the treatment of malignant tumors through the implementation tumor cell death or inhibiting the growth of tumor cells. In General offer in the invention antitumor agents applicable in the treatment of malignant tumors and other proliferative disorders, including, without limitation, breast cancer, brain cancer, skin cancer, cervical cancer, cancer of the colon and rectum, leukemia, lung cancer, melanoma, multiple myeloma, nahodkinskuju lymphoma, R is to the ovaries, pancreatic cancer, prostate cancer and stomach cancer, with named only a few. In some embodiments, the antitumor agent according to the invention is active against leukemia cells and melanoma cells and, therefore, applicable to the treatment of leukemia (e.g., myeloid, lymphocytic, promyelocytic, malacitana and lymphoblastic leukemia, or acute or chronic forms) and malignant melanomas. In other embodiments, the antitumor agent according to the invention is active against solid tumors and kill and/or inhibit the growth of cells with multidrug resistance (MDR)cells. In some embodiments, the antitumor agent according to the invention is active against malignant tumors that are resistant to other known anti-cancer agents or who were found not respond clinically to other known anti-cancer agents. In other embodiments, the antitumor agent according to the invention is active against malignant tumors that are resistant to other anticancer stabilizing microtubules agents (such as paclitaxel).

Also it will be understood that the compounds and pharmaceutical compositions proposed in this invention can be used in combination therapies, that is, connections and headlights is asepticheskie compositions can be administered simultaneously, before or after one or more other desired therapeutics or medical procedures. For a specific combination of therapeutic methods (therapeutics or procedures)that are used in a combined scheme, there is adopted the compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. Also it will be clear that the applied therapeutic methods can achieve the desired effect for the same disorder (for example, proposed in the invention compound can be administered simultaneously with other anticancer agent), or they can achieve other effects (e.g., control of any adverse effects).

For example, other therapies or anticancer agents that can be used in combination with this invention antitumor agents include surgery, radiation therapy (only a few examples are gamma rays, radiation therapy, neutron beam, radiation therapy, electron beam, proton therapy, brachytherapy, and systemic radioactive isotopes, while a few), endocrine therapy, biological response modifiers (interferons, interleukins, and tumor necrosis factor (TNF), it was also stated that the are some), hyperthermia and cryotherapy, agents to attenuate any adverse effects (e.g., antiemetics), and other approved chemotherapeutic drugs, including, but not limited to, alkylating drugs (mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide), antimetabolites (methotrexate), purine antagonists and pyrimidine antagonists (6-mercaptopurine, 5-fluorouracil, cytarabin, gemcitabine), spindle poisons division (vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel), podophyllotoxins (etoposide, irinotecan, topotecan), antibiotics (doxorubicin, bleomycin, mitomycin), nitrosoanatabine (carmustin, lomustin), inorganic ions (cisplatin, carboplatin), enzymes (asparaginase), and hormones (tamoxifen, leuprolide, flutamide, and megestrol), named only a few. For a more complete discussion of advanced methods of treatment of malignant tumors, see the list of FDA approved medicines for Oncology , Merck Manual, the seventeenth Ed. 1999, the full contents of which are incorporated in this description by reference.

In the following aspect, the invention also relates to a pharmaceutical package or kit containing one or more containers filled with one or more ingredients of the pharmaceutical compositions according to the image is the shadow, in some embodiments, it includes an additional approved therapeutic agent for use in combination therapy. Optional to such container(s) can be attached sheet in the form prescribed by the state body regulating the manufacture, use or sale of pharmaceutical products, and this piece reflects the resolution authority of the manufacture, use or sale for the introduction of man.

Equivalents

Typical examples that follow are intended to help illustrate the invention and are not intended to limit the scope of the invention, and should not be considered as such. Indeed, various modifications of the invention and many additional variations in its implementation, in addition to those shown and described in this publication will be obvious to experts in this field on the basis of the full contents of this document, including examples that follow, references to the scientific and patent literature cited in this description. In addition, it should be clear that the content of the above cited publications are included in this description by reference, to help illustrate the state of the art. The following examples contain important additional information, illustrative examples and head of the government, which can be adapted with respect to the practice of the present invention in different variants of its implementation and its equivalents.

Illustrative examples

Example 1

Synthesis of 9,10-degidro-12,13-desoxyepothilone

This example describes the synthesis of TRANS-9,10-degidro-12,13-desoxyepothilone B, 26 trifter-TRANS-9,10-degidro-12,13-desoxyepothilone B, 26 trifter-12,13-desoxyepothilone B and 12,13-desoxyepothilone B and biological testing of these compounds.

Fluorinated derivatives epothilones received and tested, taking into account increased pharmacokinetics and chemical indicators of other medicines with substituents in the form of fluorine (Ojima, I.; Inoue, T.; Chakravarty, S.; J. Fluorine Chem. 1999, 97; Newman, R. A.; Yang, J.; Finlay, M. R. V.; Cabral, F., Vourloumis, D.; Stephens, L. C.; Troncoso, P.; Wu, X.; Logothetis, C. J.; Nicolaou, K. C.; Navone, N. M. Cancer Chemother. Pharmacol. 2001, 48, 319-326; each publication included in this description by reference).

To obtain compound 2, the authors sought to take advantage of vysokogomogennogo the way, recently described in the publications of the laboratory of the authors of the invention for the synthesis of epothilone 490 (6, dehydrogenase-Epo B) as the way to dEpoB (1, scheme 3) (Biswas, K., Lin, H., Njardarson, T., Chappell, M.D., Chou, T. C., Guan, Y., Tong, W.P., He, L., Horwitz, S.B., Danishefsky, S. J. J. Am. Chem. Soc. 2002, 124 (33); 9825-9832; Rivkin, A., Njardarson, J. T., Biswas, K., Chou, T. C., Danishefsky, S. J. J. Org. Chem. 2002, 67, 7737-7740 each publication included in this description by reference). In the case of this synthesis, the authors have introduced flanking the vinyl group in compound 4 through stereospetsifichno combination of Steele's predecessor vinylidine 3 with tri-n-butylaniline. The metathesis with snapping cycle with the subsequent removal of the protection led to 6, which then turned into dEpoB (1) by regioselective reduction with diimide.

First, the focus was on the synthesis of 15 (scheme 4). Alkylation of enolate lithium 7 reported previously (Chappell, M. D.; Stachel, S. J.; Lee, C. B.; Danishefsky, S. J. Org. Lett. 2000, 2 (11), 1633-1636; included in this description by reference), iodide 8 (synthesized from the known alcohol 16 with the use of TMSI in methylene chloride) gave 9-exit 78%) and high diastereoselectivity (>25:1 de). Compound 9 in three stages made in connection 10, as shown. Attempts to enlist bromide Metalmania due to amide Weinrebe 10 were unsuccessful. Violation of this reaction was due to the presence of communication yodaiken. However, the authors were able to carry out its purpose by changing the order of these two stages of education communication C-C. Thus, after interaction with 10 vinyltrimethylsilane at the Steele can follow the accession of the Grignard reagent containing methyl, obtaining the desired ketone 11. Condensation of ketone 11 with phosphinic the home 12 with the subsequent removal of the protection triethylsilyl ether gave the fragment 13 in good yield. Etherification of the resulting 13 C1-C10 fragment acid 14 (Biswas, K.; Lin, H.; Njardarson, J. T.; Chappell, M. D., Chou, T. C., Guan, Y.; Tong, W. P., He, L.; Horwitz, S. B., Danishefsky, S. J. J. Am. Chem. Soc. 2002, 124 (33); 9825-9832; Rivkin, A.; Njardarson, J. T.; Biswas, K.; Chou, T. C.; Danishefsky, S. J. J. Org. Chem. 2002, 67, 7737-7740; publications included in this description by reference) gave the desired compound 15 with 75% yield (scheme 4).

Unfortunately, attempts to carry out the metathesis reaction with the circuit loop 15 using the catalyst of Grubbs second generation (Reviews: Grubbs, R. H.; Miller, S. J.; Fu, G. C. Acc. Chem. Res. 1995, 28, 446; Trnka, T. M.; Grubbs, R. H. Acc. Chem. Res. 2001, 34, 18; Alkene Metathesis in Organic Chemistry Ed.: Fürstner, A.; Springer, Berlin, 1998; Fürstner, A. Angew. Chem. Int. Ed. Engl. 2000, 39, 3012; Schrock, R. R. Top. Were obtained. Chem. 1998, 1, 1; each of which is incorporated in this description by reference) in methylene chloride was led mainly to an explicit dimerization of the original substance (equation 1). Given the fact that RCM works pretty well in similar conditions 5 → 6, the authors, of course, attributed the failure in the case 15 to the presence of triptorelin group C12.

It has been hypothesized that the harmful effects of constant 26 trifter-substituent on the desired reaction can be weakened by adding carbon spacer between the reaction centre RCM and triptorelin group. Accordingly, the authors have undertaken the synthesis of 19 (uravnenii) via metathesis with cycle closing connection 18, which may be triptorelin group in the context of the 17-membered cycle, containing added (1,4)-diene.

Program synthesis, aimed at 19, started with receiving the connection 21, which corresponds to O-alkyl part of the proposed by the authors of the substrate RCM (scheme 5). The authors started with Allilueva 10, this time in conditions of radical reactions, as shown (Keck, G. E.; Yates, J. B. J. Am. Chem. Soc. 1982, 104, 5829; review: Curran, D. P. Synthesis 1988, Part 1, pp.417-439; Part 2, p.489; each publication included in this description by reference). After this transformation was followed by interaction of the alkylated product with bromide Metalmania, thus giving the desired ketone 20. Condensation of this compound with a phosphine oxide 12 with the subsequent removal of the protection functions triethylsilyl ether gave 21 in good yield.

Etherification 21 C1-C10-acid fragment of 14 gave the proposed RCM precursor 18 with 75% yield (scheme 6). In this case, the metathesis reaction with the circuit loop 18 can be successfully carried out using the catalyst of Grubbs second generation in methylene chloride. As in the case of turning 5 → 6, the reaction gave exclusively the TRANS isomer 22 with the release of 57%. Finally, reductive cleavage of the protective group trichlorocarbanilide with ISOE what Itanium zinc and acetic acid, with subsequent removal of the protection of TES-ether HF-pyridine gave the desired compound 19, containing triptorelin function in C12although in the context of a number of 17-membered cycles.

A synthetic compound 19 was evaluated with respect to its cytotoxic activity. As shown in table 1-1 below, a direct comparison [17]ddEpoB (23)reported previously, with a 27-F3-[17]ddEpoB (19) showed that the new perfluorinated compound had relatively high cytotoxic efficiency.

Table 1-1
Cytotoxicity in vitro (IC50) in relation to the lines of tumor cellsand
ConnectionRF-CEM
(IC50(µm)and)
RF-CEM/VBL
(IC50(µm)and)
27-F3-[17]ddEpoB (19)0,0680,191
[17]ddEpoB (23)0,0400,126
[16]ddEpoB (6)0,0200,068
aTempl-analysis after 72 h inhibition. CCRF-CEM is a cell line T-cell acute lymphoblastic leukemia h the rights. All cell line CCRF-CEM/VBL100, CCRF-CEM/VM1and CCRF-CEM/Taxolsverkhekspressiya P-glycoprotein and show the phenotype of multidrug resistance analytical associated with MDR (Ojima, I.; Inoue, T.; Chakravarty, S.; J. Fluorine Chem. 1999, 97; Newman, R. A.; Yang, J.; Finlay, M.R.V.; Cabral, F., Vourloumis, D.; Stephens, L. C.; Troncoso, P.; Wu, X.; Logothetis, C. J.; Nicolaou, K. C.; Navone, N. M. Cancer Chemother. Pharmacol. 2001, 48, 319-326; each publication included in this description by reference).

Although triptoreline isostere substitution has little effect on the overall cytotoxic activity, preliminary studies of metabolic degradation in the plasma of mice showed that 19 is considerably more stable than the original compound 23. Exposure epothilones 19 and 23 with the plasma of nude mice and humans leads to degradation 23 within 30 minutes, whereas epothilone 19 remained mostly intact. Because pharmacokinetic problems are likely to be critical in a real application epothilones agent as a drug, the authors believe these observations are quite encouraging.

Synthesis of 26-F3-dEpoB (2) can be achieved by vysokokonkurentnoj techniques, similar to the technique used in the synthesis of 27-F3-[17]ddEpoB (19). Thus, fragments of similar complexity can serve as key building blocks (with the EMA 7). The authors suggested that acyl section 25 can serve as polipropileno domain and alkyl area 21 or 24 can be obtained, as described earlier in the introduction. The merger of the two fragments of 21 (24) and 25 can be initiated by means of esterification and completed in the subsequent metathesis with snapping cycle. Finally, cleavage of protective groups can lead to the desired analogues 28 and 29. Chemoselective recovery 9,10-olefin 28 and 29 can give dEpoB (1) and required 26-F3-12,13-disoxaril (2).

The synthesis of 1 and 2 began with obtaining acyl section 25. Ketone 30, reported previously, was subjected to aldol reaction with readily available aldehyde 31. After deprotonation and reaction "Li"-30 31, mild condensation led to a mixture of 5.3:1 Andolini products 32 and 33. The main diastereoisomer 32 were easily separated by flash chromatography and was protected as TBS-silloway ether. Hydrolysis diisopropylamino groups in the conditions of acid catalysis gave ketoaldehyde 34, defining a second stage of aldol reaction. Following earlier used to practice the method on the basis of "titanium"is a complex tert-butyl ester using the new aldehyde 34 as a partner in linking required alderny product 35 with high diastereoselectivity (dr > 20:1) and o is the house (86%). After defending C3-alcohol 35 TES-silyl was followed by removal of the protecting benzyl ether. Oxidation of the resulting primary hydroxy-group gave the corresponding aldehyde, which is then turned end-olefin by reaction of Wittig, receiving 36 with high yield. Finally, hydrolysis of complex tert-butyl methyl ether 36 using TESOTf gave acyl section 25 (82%) along with a byproduct of 37 (14%), which was converted into acyl section 38 with high yield. Spectral and chromatographic properties 38 were identical properties previously obtained substances on the basis of other programs in the laboratory of Dr. Sinha (Scripps).

The esterification of allyl alcohols 21 and 24 C1-C9acid fragment 25 gave the corresponding predecessors RCM-cyclization 26 and 27, respectively (scheme 9).

Then there was the reaction of metathesis with the closure of the loop 26, 27 and 54, using the catalyst of Grubbs second generation in toluene, which gave, as in the earlier study authors, only the TRANS-isomer 39a, 40a and 55 along with corresponding by-products 39b, 40b and 56. Finally, remove protection cyrilovich esters using HF-pyridine led to the desired compounds 28, 29 and 57. Spectral and chromatographic properties 28 were not Ident is CNAME properties previously obtained substances based programs epothilones in the laboratory of Dr. James D. White (Oregon State University). Dr. James D. White, believing that he synthesized 28 instead, however, he inadvertently got 12,13E-isomer 41, which may explain poor biological activity, which he watched. Therefore, the inventors first synthesized 28 and tested the connection with respect to its antitumor activity.

Fully synthetic 28, 29 and 2 were evaluated with respect to several types of cells, to determine their antitumor efficacy. As shown in table 1-2, all three compounds showed high cytotoxic activity against a variety of sensitive and resistant lines of tumor cells. Direct comparison 28 with dEpoB (1)reported previously, indicates that a new connection has almost three times greater efficiency.

Table 1-2
Cytotoxicity in vitro (IC50) in relation to the lines of tumor cellsand
Lines of tumor cellsIC50(µm)and
2829dEpoB (I)57
CCRF-CEM 0,00140,00350,00360,00051
CCRF-CEM/VBL1000,00650,02100,0140,0106
CCRF-CEM/Taxol0,00170,00570,00570,00073
aTempl-analysis after 72 h inhibition. CCRF-CEM is a cell line T-cell acute lymphoblastic leukemia person. All cell line CCRF-CEM/VBL100, CCRF-CEM/VM1and CCRF-CEM/Taxolsverkhekspressiya P-glycoprotein and show the phenotype of multidrug resistance-related MDR analytical (Prié, G.; Thibonnet, J.; Abarbri, M.; Duchene, A.; Parrain, J. Synlett. 1998, 839; included in this description by reference).

To increase the overall yield of the synthesis 28, 29 and 2, the authors decided to implement the RCM reaction in the absence of the substituted thiazole of olefin and, in doing so, to avoid the formation of undesirable by-product 39b and 40b. Remove protection salelologa ether 42 and 20, previously reported, gave hydroxyketone 43 and 44. Etherification of the resulting hydroxyketones 43 and 44 C1-C9acid fragment 25 Yes the Ala respective predecessors RCM-cyclization 45 and 46, respectively (scheme 10). Then carried out a metathesis reaction with the closure of the loop 45 and 46, using the catalyst of Grubbs second generation in toluene, which gave, as in the earlier study authors, only the TRANS-isomer 47 and 48 with high yields. The introduction of the rest of the triazole gave 39a, 40a and 55 with high yield. Remove protection two cyrilovich esters HF-pyridine resulted in 28 and 29. Finally, selective reduction of a C9-C10-olefin given appropriate epothilone 1 and 2. Structure 28 was strictly confirmed by its conversion into 1 with high yield. Total synthesis of 1 was significantly simplified in comparison with previously used in practice ways. Thus, the application of readily available 31 obtained from the chiral pool, of course, is a big improvement compared to the case when relying on (S)-2-methyl-4-pentenal, the synthesis of which requires intermediate chiral auxiliaries.

In the case of compound 28 in the presence of strictly confirmed the structure, the authors were surprised to find that its spectral properties do not match the properties of the connection, which was previously reported, presumably the same connection. However, when in the past, it is clear that compound 28 was not previously obtained, in deistvitel the particular whole family (E)-9,10-dehydroemetine, specified in this description, is a new class of compounds.

The study of synthetic analogues (2, 28 and 29) in cell culture showed a stronger inhibitory effect on a variety of sensitive and MDR-line tumor cells than the effects of the logged-in clinic dEpoB (1) (table 1-3). The authors note that Epo 3 (28) is the first compound 12,13-desoxyepothilone, which has significantly increased cytotoxicity compared with the cytotoxicity dEpoB (1).

Table 1-3
Cytotoxicity in vitro (IC50) in relation to the lines of tumor cellsa
ConnectionRF-CEM(C)
(µm)
With/VBL100
(µm)
With/Taxol (µm)
Are (1, dEpoB)0,00360,0160,0046
EPO 2 (2)0,00410,0800,018
EPO 3 (28)0,00090,00420,0012
EPO 4 (29)0,00350,02100,0057
aTempl-analysis after 72 h inhibition. CCRF-CEM is a cell line T-cell acute lymphoblastic leukemia person. Cell line CCRF-CEM/VBL100resistant to vinblastine, and CCRF-CEM/Taxol to Taxol.

Significant inhibition of cell growth, provided epothilone 2, 28 and 29 (Epo 2-4) on a number of different drug-resistant tumors, led to the determination of the stability of these new (E)-9,10-representatives in the blood plasma. For example, recently described (E)-10,11-degidro-dEpoB (in case 1 with CH3group at C-12) shows a very poor stability in plasma in relation to the disclosure of the lactone. It is this instability in the plasma hindered the advancement (E)-10,11-degidro-dEpoB. In contrast, exposure 2, 28 and 29 (Epo 2-4) with the plasma of the mice, the authors observed a much slower degradation of the drug in comparison with dEpoB (1), seven times. This stability represents a significant improvement from the point of view of availability of medicines, compared with dEpoB (see Fig.9).

Combining data on cytotoxicity and stability in plasma led the authors to synthesize significant amounts of 28 (Epo 3)to determine ecoeffectiveness in vivo in nude mice, bearing xenografts of human tumors. Epothilone 28 (Epo 3) showed significantly increased efficiency in the inhibition of growth of implanted tumors compared with dEpoB (see figure 10). Improved efficiency and stability of the plasma makes it possible to very significantly reduce the dose of the drug (on the order of values in the context of xenografts in the case of 28 (Epo3).

In earlier studies, the authors found that epothilone B in the form of 12,13-epoxide significantly more cytotoxic than its 12,13-detoxing (dEpoB). However, from the point of view of therapeutic index detoxication seemed to the authors is much more promising. Recently, the authors reported the synthesis of (E)-9,10-degidro-12,13-desoxyepothilone B (28) using stereoselective metathesis with snapping cycle. The authors showed that the introduction of E-9,10-unsaturation in the context of the usual Z-12,13-olefin (see compound 1) leads to a strong increase efficiency in vitro. Essentially, this can be transferred to conditions in vivo in mice with xenotransplantation. In addition, the connection 28 has a significant pharmaceutical benefits compared to dEpoB (1). This makes it possible to reduce the dose levels 28, compared with 1 in experiments with xenografts, which reduce the order of the values.

Accordingly, the authors interest was the echo, can the introduction of C9-C10-olefin in epothilone B (51, EpoB) change its biological profile in the same direction.

Epoxidation 28 2,2'-dimethyldioxirane (DMDO) occurred with high chemoselectivity the more substituted C12-C13-olefin, giving exit 87% ratio 1:2,6 (E)-9,10-dehydroemetine B (49) and its diastereoisomer (50). The stereochemistry of the epoxides was determined by selective recovery diimide C9-C10-double bonds. The study of spectral properties of these products revealed that the minor product (49) is dEpoB. Preferred α-epoxidation in the case 28 is completely different from the highly stereoselective epoxidation dEpoB, which occurs with β-hand, leading to EpoB (Meng, D.; Bertinato, P.; Balog, A.; Su, D.-S.; Kamenecka, T.; Sorensen, E. J.; Danishefsky, S. J. J. Am. Chem. Soc. 1997, 119, 10073; included in this description by reference).

(E)-9,10-dehydroemetine B (51) were evaluated in relation to many different types of cells, to determine their antitumor efficacy. As shown in table 1-4, (E)-9,10-dehydroemetine B (49) shows high cytotoxic activity against sensitive and resistant lines of tumor cells. Direct comparison 49 and EpoB (51) shows that the new analogue has almost 3 times more efficient than EpoB (51), making it one of the most e is effective analogues epothilone, reported to date. Interestingly, a series of α-epoxides (50, 52) shows much less activity than EpoB (51). The graph below shows data in vivo studies of compounds 49.

Table 1-4
Cytotoxicity in vitro (IC50) in relation to the lines of tumor cellsa
ConnectionRF-CEMCRF-CEM/VBLCRF-CEM/Taxol
1 dEpoB)0,00360,0160,0046
280,00090,00420,0012
51 (EpoB)0,000620,00370,0011
490,000230,000320,00042
500,01340,09590,0802
520,0830,4519 0,1507
aTempl-analysis after 72 h inhibition. CCRF-CEM is a cell line T-cell acute lymphoblastic leukemia person. All cell line CCRF-CEM/VBL and CCRF-CEM/Taxol sverkhekspressiya P-glycoprotein and show the phenotype of multidrug resistance-related MDR analytical.

Therapeutic effect of 9,10-de-H-EpoB nude mice bearing xenograft MX-1 (6-h/-infusion, n=4).

In conclusion, the above represents an efficient stereoselective total synthesis of 28 (Epo 3) and after the site-selective recovery diimide dEpoB (1) as such. Described in this description of technique is then directly applied to obtain the relevant trifter analogues 2 and 29 (Epo 4). In addition, epoxidation 28 gave 49 and 50 which, when the site-selective recovery diimide gave epothilone B (51) and 52. The above data indicate the appearance of the most promising new family of anticancer drugs that should be assessed in relation to the possible ways in this case, advance to the conditions of clinical trials in humans. In addition, a new method of synthesis involves significant practical improvements total synthesis of dEpoB and epothilone B.

Experimental studies

General procedures:Reagents obtained from commercial suppliers were used without further purification unless otherwise stated. The following solvents were obtained from the anhydrous solvent system (passed through a pre-filled column of alumina) and used without further drying: tetrahydrofuran, methylene chloride, diethyl ether, benzene and toluene. All are sensitive to the atmosphere and water of reaction was carried out in a flame dried glassware under a positive pressure of pre-purified argon gas. NMR spectra (1H and13C) were recorded on a Bruker AMX-400 MHz or Bruker Advance DRX-500 MHz, which are listed separately, using as a reference CDCl3(7,27 ppm for1H and of 77.0 ppm for13C). Infrared spectra (IR) were obtained on a spectrometer Perkin-Elmer FT-IR model 1600. Optical rotations were obtained on a digital polarimeter JASCO model DIP-370 at 22±2°C. Analytical thin-layer chromatography was carried out on plates of E. Merck silica gel 60 F254. Compounds that were not UV active, visualized by dipping the plates in a solution of cerium molybdate-ammonium or para-anisaldehyde and heating. Chromatography on silica gel was performed using the indicated solvent on silica gel Davisil® grade 1740, type 60A, 170-400 mesh).

Acronymy reduction

TES, triethylsilyl; TBS, dimethyltrimethylene; EDCI, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; HF-PY, hydrogen fluoride in pyridine; DMAP, 4-N,N-dimethylaminopyridine; DHM, dichloromethane; DMF, N,N-dimethylformamide; THF, tetrahydrofuran.

Compound 32:To a solution of freshly prepared LDA (11.6 mmol) in THF (25 mmol) was added dropwise a solution of ketone 30 (2,40 g, 10.4 mmol) in THF (6.8 ml) at -78°C. After stirring at -40°C for 0.5 hour the mixture was cooled to -90°C. was added dropwise a solution of aldehyde 31 (1,38 g, 7,72 mmol) in THF (6.8 ml). After stirring at -90°C for 35 min the reaction was suppressed saturated aqueous NH4Cl (15 ml) and was extracted with EtOAc (50 ml × 3). The combined organic extracts were dried over Na2SO4and concentrated. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 15:1 to 12:1) gave 32 (2,09 g, 66%) and isomer 33 (0.39 g, 12%), both as yellow oils. 32: [α]D2513,1 (c 1,22, CHCl3); IR (film) ν 3494, 2972, 2932, 1708, 1454, 1380, 1329, 1120, 1038, 998, 734 cm-1;1H NMR (400 MHz, CDCl3): δ of 0.98 (3H, d, J=6,9 Hz)of 1.06 (3H, d, J=6.9 Hz), 1,10 (3H, d, J=6,1 Hz)to 1.14 (3H, d, J=6,9 Hz)and 1.15 (3H, c), of 1.17 (3H, d, J=6.2 Hz), of 1.18 (3H, c), of 1.20 (3H, d, J=6.2 Hz), 1,81-of 1.92 (1H, m), 3.33 and (1H, CVD, J=7,0, 2,2, Hz), 3,51 (1H, DD, J=8,9, and 6.3 Hz), to 3.64 (1H, d, J=1,8 Hz), 3,66-3,71 (2H, m), 3,78-3,86 (2H, m), 4,51 (1H, d, J=12.0 Hz), of 4.54 (1H, d, J=12.0 Hz), 4,58 (1H, c), 7,25-to 7.35 (5H, m);13With NMR (100 MHz, DCl3) δ 10,0, 14,3, 20,5, 21,3, 21,9, 22,5, 2,5, 23,6, 36,4, 42,1, 54,1, 69,8, 71,2, 72,8, 73,3, 73,4, 103,8, 127,6, 127,7 (2C), RUR 128.5 (2C), 138,9, 221,6; MS-HP (ESI) calculated for C24H40About5Na [M+Na+] that amount to 431,3 found 431,4.

Compound 32a(not shown): To a cooled (-40°C) solution of alcohol 32 (1.01 g, 2,47 mmol) and 2,6-lutidine (69 μl, to 5.93 mmol) was added TBSOTf (681 μl, 3.00 mmol) and the mixture was heated to -20°C for 3.5 hours. The reaction extinguished saturated aqueous NaHCO3(10 ml). After extraction with hexane (50 ml × 3) and the combined organic extracts were dried over Na2SO4and concentrated. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 50:1) gave 32a (1,25 g, 2,39 mmol, 97%) as a colourless oil; [α]D25-19,7 (0,58, Cl3); IR (film) ν 2966, 2931, 1696, 1455, 1378, 1320, 1255, 1091, 1044, 991, 873, 838, 773 cm-1;1H NMR (400 MHz, CDCl3) δ of 0.08 (6H, c)to 0.89 (9H, c)of 0.99 (3H, d, J=7,0 Hz), was 1.04 (3H, d, J=7,0 Hz)of 1.07 (3H, d, J=7,0 Hz)of 1.07 (3H, c)to 1.14 (3H, d, J=6,1 Hz)of 1.17 (3H, c), of 1.17 (3H, d, J=6.0 Hz), of 1.20 (3H, d, J=6.2 Hz), 1,76-of 1.85 (1H, m), 3,21 (1H, DD, J=9,2, 7,3 Hz), 3,32 (1H, Quint, J=7.4 Hz), 3,62 (1H, DD, J=9,2, 5.7 Hz), 3,78-of 3.85 (2H, m), a 3.87 (1H, DD, J=7,7, 2.0 Hz), 4,46 (1H, d, J=12.1 Hz), 4,50 (1H, d, J=12.1 Hz), 4,73 (1H, c), 7,24-7,37 (5H, m);13With NMR (100 MHz, DCl3) δ -3,6, -3,3, 15,6, 16,8, 18,7, 18,8, 21,8, 22,1, 22,5, 23,5, 23,7, 26,4 (3C), 39,0, 46,2, 54,0, 69,7, 70,9, 72,1, 73,4, 76,7, 103,1, 127,6, 127,8 (2C), RUR 128.5 (2C), 139,0, 218,9; MS-HP (ESI) calculated for C30H54About5SiNa [M+Na+] 545,4 found 545,4.

Connect the s 34: The mixture 32a (3.03 g, 5,79 mmol)and p-TsOH·H2O (286 mg) in aqueous THF (64 ml, THF/H2O = 4:1) was boiled under reflux for 6.5 hours. The reaction mixture was cooled to CT and was poured into saturated aqueous NaHCO3(25 ml). After extraction with EtOAc (100 ml + 50 ml × 2) and the combined organic layers were washed with saturated salt solution, dried over Na2SO4and concentrated. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 50:1 to 30:1) gave 34 (2.37 g, 5,64 mmol, 98%) as a colorless oil: [α]D25-25,8 (0,515, l3); IR (film) ν 2955, 2931, 1731, 1696, 1455, 1360, 1255, 1091, 1026, 873, 826, 767 cm-1;1H NMR (400 MHz, CDCl3) δ 0,06 (3H, c), of 0.07 (3H, c), of 0.90 (9H, c)of 0.95 (3H, d, J=7,1 Hz)of 1.03 (3H, d, J=7.0 Hz), 1.28 (in, 3H, c), of 1.33 (3H, c), 1,73-to 1.82 (1H, m), and 3.16 (1H, DD, J=9,2, 6,1 Hz), or 3.28 (1H, Quint, J=7,3 Hz), 3,55 (1H, DD, J=9,2, 6,7 Hz), 3,91 (1H, DD, J=7,8, and 2.1 Hz), 4,46 (2H, c), 7,27 and 7.36 (5H, m), 9,58 (1H, c);13With NMR (100 MHz, DCl3) δ -3,6, -3,5, 15,7, 16,3, 18,6, 19,8, 20,1, 26,3 (3C), 39,1, 47,0, 61,1, 71,9, 73,4, 75,8, 127,7, 128,0 (2C), RUR 128.5 (2C), 138,6, 201,3, 213,3; MS-HP (ESI) calculated for C24H40About4SiNa [M+Na+] 443,3 found 443,2.

Compound 35:To a solution of freshly prepared LDA (18 ml of 0.5 M solution in Et2O, 9.0 mmol) in Et2O (20 ml) was added tert-butyl acetate (1,16 ml, 8,61 mmol) at -78°C. After stirring for 50 min dropwise via syringe pump was added CpTiCl(OR)2(100 ml of 0.1 M solution in Etsub> 2O, 10.0 mmol) for 65 minutes After stirring for 20 min the reaction mixture was heated to -30°C, was stirred for 50 min and again cooled to -78°C. dropwise over 10 min was added a solution of 34 (2,42 g of 5.75 mmol) in Et2O (9 ml) and the resulting mixture was stirred at -78°C. After stirring for 2 h the reaction was suppressed aqueous THF (5 M H2O, 37 ml) and stirred at RT for 2 hours. After adding water (40 ml) and the mixture was stirred for 1 hour. The precipitate was filtered on celite (flushing Et2O) and the filtrate was washed with water (40 ml). The aqueous layer was extracted with Et2O (100 ml × 2) and the combined organic layers were washed with saturated salt solution (40 ml), dried over Na2SO4and concentrated. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 10:1) gave 35 (2.65 g, 4,94 mmol, 86%) as a pale yellow oil; [α]D25-20,3 (1,0, l3); IR (film) ν 3523, 2957, 2930, 2856, 1732, 1700, 1472, 1368, 1252, 1152, 1091, 1042, 986, 834, 774 cm-1;1H NMR (400 MHz, CDCl3) δ 0,07 (3H, c), of 0.07 (3H, c), of 0.90 (9H, c)of 0.99 (3H, d, J=7,0 Hz)of 1.07 (3H, d, J=7,0 Hz), 1,10 (3H, c)to 1.14 (3H, c), of 1.47 (9H, c), 1,77 of-1.83 (1H, m), and 2.26 (1H, DD, J=16,0, 10,0 Hz), 2,34 (1H, DD, J=15,9 with 2.7 Hz), 3,23 (1H, DD, J=9,2, 7,1 Hz), 3,35 (1H, d, J=2.7 Hz, -OH), to 3.36 (1H, Quint, J=7.0 Hz), 3,61 (1H, DD, J=9,2, 5,9 Hz), 3,88 (1H, DD, J=7,6, 2.0 Hz), 4,17 (1H, dt, J=10,0, 2.7 Hz), 4,48 (2H, c), 7,27 and 7.36 (5H, m);13With NMR (100 MHz, DCl3) δ is 3.5, -3,4, 16,3, 167, 18,7, 20,1, 21,6 of 26.4 (3C), and 28.3 (3C), 38,0, 39,1, 45,8, 51,8, 72,2, 72,9, 73,5, 76,7, 81,4, 127,7, 128,0 (2C), RUR 128.5 (2C), 138,8, 172,7, 219,6; MS-HP (ESI) calculated for C30H52About6SiNa [M+Na+] 559,3 found 559,4.

Compound 35a (not shown):To a mixture of alcohol 35 (10.2 g, to 18.9 mmol) and imidazole (2.70 g, and 39.7 mmol) in DMF (25 ml) was added TESCl (3,3 ml of 19.8 mmol) at 0°C and the mixture was stirred at RT for 2 hours. The reaction extinguished saturated aqueous NaHCO3(50 ml). After extraction with hexane (500 ml + 120 ml × 2) and the combined organic extracts are then washed with water (30 ml × 2) and saturated salt solution (30 ml), dried over Na2SO4and concentrated. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 40:1) gave 35a (12.1 g, 18.5 mmol, 98%) as a colorless oil: [α]D25-38,0 (0,46, l3); IR (film) ν 2955, 2877, 1733, 1697, 1456, 1367, 1298, 1251, 1155, 1099, 988, 835, 742 cm-1;1H NMR (400 MHz, CDCl3) δ of 0.05 (6H, c)of 0.57 and 0.68 (6H, m)to 0.89 (9H, c)of 0.95 (9H, t, J=7.9 Hz), 0,99 (3H, d, J=7,0 Hz)of 1.02 (3H, d, J=6.8 Hz), 1.04 million (3H, c), of 1.18 (3H, c), a 1.45 (9H, c), 1.70 to to 1.79 (1H, m)of 2.16 (1H, DD, J=17,0, a 7.0 Hz), 2.40 a (1H, DD, J=17,0, 3.1 Hz), up 3.22 (1H, DD, J=9,1, 7.5 Hz), and 3.31 (1H, Quint, J=6.9 Hz), 3,61 (1H, DD, J=9,1, a 5.4 Hz), 3,83 (1H, DD, J=7,3, 2.3 Hz), 4,30 (1H, DD, J=6,9, 3.1 Hz), 4,48 (2H, c), 7,27 and 7.36 (5H, m);13With NMR (100 MHz, DCl3) δ is 3.5, -3,4, 5,3 (3C), and 7.3 (3C), 15,3, 16,9, 18,7, 20,1, 23,4, 26,4 (3C), and 28.3 (3C), 39,1, 41,1, 46,2, 53,4, 72,2, 73,4, 74,3, 76,7, 80,6, 127,6, 127,9 (2C), RUR 128.5 (2C), 138,9, which is 171,5, 218,4; MS-HP (ESI) calculated for C36 H66About6Si2Na [M+Na+] 673,4 found to 673.5.

Compound 35b (not shown):To a stirred solution of 35a (4,37 g, 6,72 mmol) in THF (67 ml) was added Pd/C (purchased from Acros, 10% wt., 437 mg) and the mixture was stirred in an atmosphere of H2. After stirring for 2.2 hours the mixture was filtered through a pad celite, which were washed in THF (120 ml). The filtrate was concentrated and purified flash chromatography on a column (SiO2, hexane/EtOAc = 30:1 to 10:1), receiving 35b (3,53 g, 6,28 mmol, 94%) as a colourless oil;

[α]D25-16,1 (0,62, l3); IR (film) ν 3543, 2956, 1732, 1696, 1472, 1368, 1299, 1252, 1155, 1100, 988, 837, 775, 742 cm-1;1H NMR (400 MHz, CDCl3) δ of 0.10 (3H, c), 0,12 (3H, c), of 0.60 and 0.68 (6H, m)of 0.93 (9H, c)to 0.96 (9H, t, J=8.0 Hz), 0,99 (3H, d, J=7,1 Hz), 1,10 (3H, d, J=6,9 Hz)to 1.14 (3H, c), of 1.20 (3H, c), a 1.45 (9H, c), 1,46-of 1.55 (1H, m), 2,21 (1H, DD, J=17,2, 7,1 Hz), 2,39 (1H, DD, J=17,2, 2,8 Hz), of 2.54 (1H, t, J=5.8 Hz, HE), 3,30 (1H, Quint, J=6.9 Hz), to 3.58 (1H, dt, J=11,5, 5,5 Hz), 3,66 (1H, dt, J=11,3, 5,4 Hz)to 3.92 (1H, DD, J=8.0 a, 2,1 Hz), 4,32 (1H, DD, J=7,1, 2,9 Hz);13With NMR (100 MHz, DCl3) δ -3,6, is 3.5, 5,3 (3C), 7,2 (3C), 16,0, 16,1, 18,6, 20,0, 23,4, 26,4 (3C), AND 28.3 (3C), 40,0, 40,9, 46,9, 53,7, 64,8, 73,3, 78,1, 80,9, 171,7, 218,5; MS-HP (ESI) calculated for C29H60About6Si2Na [M+Na+] 583,4 found 583,5.

Compound 35c (not shown):To a stirred mixture of alcohol 35b (3,53 g, 6,28 mmol) and powdered MS4A (svezheasfaltirovannaya, 2.50 g)in CH 2Cl2(32 ml) was added NMO (1,17 g, 10.0 mmol), then TPAP (132 mg, 0,377 mmol). After stirring at RT for 35 min the mixture was filtered through a column of silica gel (hexane/Et2O = 8:1), receiving 35c (3,34 g, 5,98 mmol, 95%) as a colourless oil; [α]D25-69,6 (with 0.25, l3); IR (film) ν 2955, 2878, 1732, 1696, 1472, 1368, 1253, 1155, 1097, 989, 837 cm-1;1H NMR (400 MHz, CDCl3) δ 0,09 (3H, c), 0,10 (3H, c), of 0.59 and 0.68 (6N, m), 0,89 (N, c), 0,95 (N, t, J=8.0 Hz), a 1.08 (3H, c), is 1.11 (3H, d, J=6,9 Hz)to 1.14 (3H, d, J=7,1 Hz), 1,24 (3H, c), 1,45 (N, c), 2,19 (1H, DD, J=17,0, 6,7 Hz), 2,33 (1H, cut, J=7,1, 2.2 Hz), is 2.41 (1H, DD, J=17,0, and 3.3 Hz), or 3.28 (1H, Quint, J=7.5 Hz), 4,07 (1H, DD, J=7,9, 2.2 Hz), 4,32 (1H, DD, J=6,7, and 3.2 Hz), 9,74 (1H, d, J=2.0 Hz);13With NMR (100 MHz, DCl3) δ -3,8, is 3.5, 5,3 (3C), 7,2 (3C), 12,6, 15,6, 18,5, 20,5, 23,3, 26,2 (3C), AND 28.3 (3C), 41,1, 46,9, 51,1, 53,5, 74,0, 76,5, 80,7, 171,1, 204,3, 218,0 MS-HP (ESI) calculated for C29H58About6Si2Na [M+Na+] 581,3 found 581,3.

The connection 36:MePPh3I (2,56 g, 7,18 mmol) in THF (40,0 ml) was treated with t-BuOK (6,57 ml of 1.0 M solution in THF; to 6.57 mmol) at 0°C. After stirring at 0°C for 20 min, the resulting suspension was cooled to -78°C was added a solution of aldehyde 35 (3,34 g, 5,98 mmol) in THF (14 ml). After stirring at -78°C for 15 min and the mixture was stirred at 0°C for 15 min and at RT for 15 min. the Reaction was suppressed saturated aqueous NH4Cl (20 ml) and was extracted with Et2O (120 ml + 50 ml × ). The combined organic extracts were washed with saturated salt solution (20 ml), dried over Na2SO4and concentrated. The residue was purified flash chromatography on a column (SiO2~80 g, hexane/Et2O = 40:1)to give 36 (125,3 mg, 0,225 mmol, 78%) as a colourless oil; [α]D25-33,6 (0,250, l3); IR (film) ν 2956, 2878, 1733, 1696, 1472, 1367,1299, 1253, 1156, 1100, 988, 837, 774 cm-1;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), and 0.08 (3H, c), of 0.60 and 0.68 (6H, m)of 0.93 (9H, c)to 0.96 (9H, t, J=8.0 Hz), was 1.04 (6H, d, J=7,0 Hz)of 1.09 (3H, c), of 1.20 (3H, c), a 1.45 (9H, c), 2,08-to 2.15 (1H, m)to 2.29 (1H, DD, J=17,0, 7,0 Hz), to 2.41 (1H, DD, J=17,0, 3.1 Hz), is 3.08 (1H, Quint, J=7.0 Hz), a-3.84 (1H, DD, J=7,0, 2,1 Hz), 4,32 (1H, DD, J=7,0, 3.1 Hz), 5,02 (1H, DD, J=17,9, 1.0 Hz), is 5.06 (1H, DD, J=10,5, 1.0 Hz), to 5.93 (1H, DDD, J=17,9, 10,5, 7,7 Hz);13With NMR (100 MHz, DCl3) δ -3,6, -3,3, 5,4 (3C), 7,2 (3C), 15,2, 18,7, 19,0, 20,2, 23,6, 26,4 (3C), AND 28.3 (3C), 41,1, 43,8, 46,4, 53,5, 73,9, 76,6, 80,6, 115,5, 140,2, 171,5, 218,5; MS-HP (ESI) calculated for C30H60About5Si2Na [M+Na+] 579,4 found 579,4.

Compound 25:The complex solution of tert-butyl methyl ether 36 (4,87 g, a total of 8.74 mmol) and 2,6-lutidine (fresh, 4,1 ml of 35.0 mmol) in CH2Cl2(58 ml) was added TESOTf (4,0 ml, 17.5 mmol) at 0°C. After stirring at 0°C for 25 min and the mixture was stirred at RT for 3.2 hours. The mixture was diluted with Et2O (600 ml), sequentially washed with 5% aqueous KHSO4(60 ml × 2) and saturated salt solution (60 ml), dried over N 2SO4and concentrated. The residue was dried under high vacuum for 1.5 hours, receiving the crude acid 25 (6,30 g, contaminated TESOH). The crude product (6,30 g) was dissolved in aqueous THF (87,5 ml, THF/H2O = 6:1) and treated with saturated aqueous NaHCO3(12.5 ml). After stirring at RT for 20 min, the resulting suspension was diluted with Et2O (500 ml) and acidified water 5% KHSO4(55 ml). After separation of the layers the aqueous layer was extracted with Et2O (100 ml × 2) and the combined organic layers were washed with saturated salt solution (50 ml × 2), dried over Na2SO4and concentrated. The residue was dried under high vacuum over night, receiving the crude acid (ceiling of 5.60 g, contaminated TESOH) as a colourless oil, which was used for next reaction without further purification. To characterize the purified flash chromatography on a column of silica gel, elwira a mixture of hexane/EtOAc = 4/1. [α]D25-30,7 (0,985, l3); IR (film) ν 2956, 2936, 2879, 1712, 1472, 1417, 1303, 1253, 1107, 1046, 1003, 988, 872, 837, 775, 741 cm-1;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), and 0.09 (3H, c), 0,59 is 0.67 (6H, m)of 0.93 (9H, c)to 0.96 (9H, t, J=8.1 Hz), of 1.05 (3H, d, J=7,0 Hz)of 1.05 (3H, d, J=7,0 Hz)of 1.20 (3H, c)to 1.21 (3H, c), of 2.06 and 2.13 (1H, m), 2,34 (1H, DD, J=16,4 that 7.4 Hz), 2,50 (1H, DD, J=16,4, 3.0 Hz), 3,06 (1H, Quint, J=7,3 Hz), a 3.87 (1H, DD, J=7,5, 1,8 Hz), and 4.40 (1H, DD, J=7,3, 2,9 Hz), free 5.01 (1H, DD, J=18,0, and 0.9 Hz), 5,07 (1H, DD, J=10,4, 1.2 Hz), 593 (1H, DDD, J=18,0, 10,4, 7,8 Hz);13With NMR (100 MHz, DCl3) δ -3,6, -3,3, 5,3 (3C), and 7.1 (3C), 15,6, 18,7, 19,1, 19,2, 24,1, 26,4 (3C), 39,8, 43,6, 46,4, 53,5, 73,7, 76,6, 115,6, 140,0, 177,9, 218,7; MS-HP (ESI) calculated for C26H52About5Si2Na [M+Na+] 523,3 found 522,9.

Compound 45:3-O-TES-6-O-TBS-protected acid 25 was dried by azeotropic distillation with benzene. Dry alcohol 43 (200 mg, 1,19 mmol) was dissolved in DHM (10 ml) and cooled to 0°C and was added solid DMAP (167 mg, 1.37 mmol) and solid EDCI (261 mg, 1.37 mmol). After stirring the reaction mixture at 0°C for 15 min was added dropwise a solution of acid 25 (425 mg, 0.85 mmol) in DHM (2 ml). The cooling bath was removed and stirring continued for another 2 hours. The crude reaction mixture was diluted DHM (10 ml) and was purified by chromatography on silica gel using 10% EtOAc/hexane as eluent, obtaining the ester 45 (380 mg, yield 81%, two stages, starting with 36) in the form of a clear oil: [α]D-15,1 (c 1,2, CDCl3); IR (pure substance) 2955, 2932, 2877, 1743, 1732, 1694, 1474, 1461, 1417, 1380, 1360, 1295, 1252, 1169, 1094, 1043, 988,3, 912,9, 871,4, 836,5, 774,8, 741,6 cm-1;1H NMR (500 MHz, CDCl3) 0,08 (3H, c), and 0.08 (3H, c), of 0.60 and 0.68 (6H, m)0,93 (N, c), 0,95 (N, t, J=8.0 Hz), was 1.04 (3H, d, J=6.9 Hz), of 1.05 (3H, d, J=6.9 Hz), 1,10 (3H, c), 1,25 (3H, c), was 1.69 (3H, c), 2,08-of 2.15 (2H, m)of 2.16 (3H c), of 2.38 (1H, DD, J=17,0,7,0 Hz), 2,48 (2H, t, J=6.5 Hz), to 2.57 (1H, DD, J=17,0, 2.7 Hz), 2,71 was 2.76 (2H, m), of 3.07 (1H, Quint, J=7.0 Hz), 3,83 (1H, q, j =7.2 Hz), 4,36 (1H, DD, J=7,0, 2.7 Hz), equal to 4.97 is 5.07 (4H, m), 5,19 (1H, t, J=7,0), 5,73 (1H, TD, J=15,4, 5,9 Hz), of 5.92 (1H, DD, J=15,7, 8.0 Hz);13With NMR (500 MHz, DCl3) δ 218,4, 205,4, 172,1, 140,1, 137,4, 135,4, 119,1, 115,8, 115,6, 78,7, 76,5, 73,9, 53,3, 46,3, 43,7, 39,6, 36,6, 29,2, 26,7, 26,4, 23,8, 23,7, 19,9, 18,9, 18,7, 15,4, 7,06, 5,30, -3,29, -3,62; MS-HP (ESI) calculated for C36H66About6Si2Na [M+Na+] 673,4 found to 673.5.

Compound 47:To a solution of compound 45 (20 mg, 0,031 mmol) in anhydrous toluene (60 ml) by boiling under reflux for one portion was added a solution of the dichloride tricyclohexylphosphine[1,3-bis(2,4,6-trimetilfenil)-4,5-dihydroimidazole-2-ilidene][benzylidene]ruthenium (IV) (5.2 mg, 0,0061 mmol) in anhydrous toluene (2 ml) and the mixture was heated for 10 minutes. The reaction mixture was immediately cooled in a bath with ice, was applied onto silica and purified using chromatography on silica gel using 4-10% gradient EtOAc/pentane as eluent to obtain compound 47 (15 mg, yield 78%) as an oil: [α] -28,6 (1,2, l3); IR (pure substance) 2955, 2933, 2878, 1745, 1731, 1695, 1471, 1462, 1380, 1361, 1251, 1159, 1104, 1080, 1019, 985,0, 876,1, 835,5, 774,7, 743,1, 670,1 cm-1;1H NMR (500 MHz, CDCl3) of 0.07 (3H, c), 0,10 (3H, c), of 0.59 and 0.68 (6H, m)of 0.91 (9H, t, J=8.0 Hz), of 0.93 (9H, c), was 1.04 (3H, d, J=7,0 Hz), 1,10 (3H, c), is 1.11 (3H, d, J=7,0 Hz)of 1.17 (3H, c), 1,71 (3H, c), of 2.21 (3H, c), 2,27 of-2.32 (1H), of 2.38 (1H, DD, J=14,6, 6,8 Hz), of 2.51-2,61 (2H, m), 2.57 m (1H, DD, J=15,5, and 3.3 Hz), 2.93 which is 3.1 (3H, m), of 3.94 (1H, d, J=8.5 Hz), 4,28 (1H, DD, J=8,6, 3.0 Hz), 5,04 (1H, DD, J=8,7, 2,), 5,16 (1H, t, J=7,5), 5,73 (1H,tDD, J=12,8, 9,94 and 6.9 Hz), of 5.92 (1H, DDD, J=18,0, 10,3, 7,8 Hz);13With NMR (125 MHz, DCl3) δ 215,9, 204,8, 171,3, 140,0, 132,7, 129,2, 118,6, 79,1, 78,2, 75,4, 54,0, 48,2, 41,7, 40,3, 35,0, 29,2, 26,6, 26,5, 23,5, 22,8, 20,6,18,8, 17,5, 14,3, 7,19, 5,53, -3,36; MS-HP (ESI) calculated for C34H62About6Si2645,4 found 645,4 (M+Na+).

Compound 39a:To a solution of Wittig reagent (19.1 mg, 54,7 mmol) in THF (0.4 ml) was added KHMDS (109 μl 0.5 M solution in toluene, 54,7 mmol) at 0°C. the Mixture was stirred at 0°C for 0.5 hour and then was cooled to -78°C. To the mixture was added dropwise a solution of ketone 47 (5.7 mg, 9,12 mmol) in THF (0.3 ml) and the resulting mixture was allowed to warm to -20°C for 1.5 hour. The reaction extinguished saturated aqueous NH4Cl (2 ml) and was extracted with EtOAc (7 ml × 3). The combined organic layers were dried over Na2SO4and concentrated. The residue was purified flash chromatography on a column (SiO2, hexane/Et2O = 10:1)to give 5.6 mg inseparable mixture of E/Z-olefins (E/Z = 9:1). The mixture was purified preparative TLC (hexane/Et2O = 4:1)to give pure 39a (5.0 mg, of 6.96 mmol, 76%) as a colourless oil; [α]D25-41,5 (0,715, l3); IR (film) ν 2955, 2884, 1737, 1690, 1467, 1378, 1249, 1179, 1102, 1014, 979, 879, 826, 773 cm-1;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), 0,12 (3H, c), or 0.57 (6H, q, J=7,8 Hz)to 0.89 (9H, t, J=8.0 Hz), of 0.93 (9H, c), was 1.04 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), 1,17 3H, d, J=7,1 Hz), 1,68 (3H, c), of 2.15 (3H, d, J=0.8 Hz), 2,14-of 2.27 (2H, m), of 2.45 (1H, DD, J=14,0, 4,8 Hz), 2,50 (1H, DD, J=14,9, and 3.2 Hz), 2,64-to 2.74 (2H, m), of 2.72 (3H, c), to 3.02 (1H, Quint, J=7.0 Hz), 3,10 (1H, DD, J=14.4V, 7,3 Hz), of 3.96 (1H, d, J=8.7 Hz), 4,43 (1H, DD, J=8,3, 2,9 Hz), with 5.22 (1H, DD, J=9,8, 5.7 Hz), 5,33-5,42 (2H, m), 5,69 (1H, DD, J=15,8, 8,2 Hz), to 6.57 (1H, c), of 6.96 (1H, c);13With NMR (100 MHz, DCl3) δ -3,3, -3,2, 5,6 (3C), and 7.1 (3C), 15,0, 17,2, 18,8, 19,4, 21,4, 21,7, 23,8, 24,3, 26,5 (3C), 33,2, 35,6, 41,3, 41,8, 48,2, 54,0, 74,4, 77,4, 79,3, 116,4, 120,5, 121,0, 129,3, 132,1, 137,8, 138,0, 152,7, 164,8, 170,7, 216,8; MS-HP (ESI) calculated for C39H68NO5SSi2[M+H+] 718,4 found 718,3.

Compound 28 (Epo 3):To a solution of 39a (298,8 mg, 0,416 mmol) in THF (6.5 ml) was added HF·pyridine (3.2 ml) at 0°C and the mixture was stirred at RT for 3 hours. The reaction was suppressed by adding dropwise TMSOMe (30 ml) at 0°C. After concentration and drying under high vacuum the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 1:1)to give 28 (196,6 mg, 0,402 mmol, 97%) as a white solid; [α]D25-96,6 (0,235, l3); IR (film) ν 3502, 2970, 2927, 1733, 1685, 1506, 1456, 1375, 1251, 1152, 1040, 977 cm-1;1H NMR (400 MHz, CDCl3) δ of 1.06 (3H, c), is 1.11 (3H, d, J=7,0 Hz)to 1.22 (3H, d, J=6.8 Hz), 1.28 (in, 3H, c), 1,72 (3H, c), 2,10 (3H, c), 2,31-to 2.40 (2H, m), 2,43 (1H, DD, J=16,0, and 3.7 Hz), 2.49 USD (1H, DD, J=16,0, 9,2 Hz), 2,55 of 2.68 (2H, m), a 2.71 (3H, c), 2,98 (1H, DD, J=14,4, 6.4 Hz), and 3.16 (1H, Quint, J=6.2 Hz), 3,76 (1H, DD, J=5,9, and 3.2 Hz), 4,30 (1H, DD, J=9,2, and 3.7 Hz), 5,18 (1H, ushort, J=7,3 Hz), 5,32 (1H, DD, J=8,4, 2,5 Hz), 5,63 (1H, DD, J=15,7, 6.4 Hz), the ceiling of 5.60 (1H, DDD, =15,7, 6,9, 5,1 Hz), 6,60 (1H, c), 6,98 (1H, c);13With NMR (100 MHz, DCl3) δ 15,1, 16,0, 17,7, 19,2, 19,5, 22,5, 23,6, 32,0, 35,0, 39,6, 40,3, 44,8, 53,3, 71,8, 75,6, 78,3, 116,1, 119,6, 120,5, 129,9, 131,3, 137,5, 138,2, 152,2, 165,0, 170,7, 218,8; MS-HP (ESI) calculated for C27H40NO5S [M+H+] 490,3 found 490,2.

dEpoB (1, Epo 1):To a solution of 28 (1.2 mg, 2.5 µmol) and NHNH2(29.3 mg, 98 μmol) in ClCH2CH2Cl (0.7 ml) at 50°C was added Et3N (13,7 μl, 98 mmol). The reaction was controlled by VETCH (hexane/EtOAc/CH2Cl2= 1/1/2). After stirring for 7 h, the mixture was cooled to CT, diluted with EtOAc and filtered through a bed of silica gel, which is washed with EtOAc. After concentration the residue was purified preparative TLC (hexane/EtOAc/CH2Cl2= 1/1/2), obtaining 1 (1.1 mg, 2.2 mmol, 91%) as a white solid. Spectral data for 1 were identical to the data described for dEpoB.

Compound 27:Before the reaction the acid 25 and alcohol 24 was subjected to azeotropic distillation with dry benzene (5 ml × 2) and dried under high vacuum. To a solution of alcohol 24 (639 mg, 2,63 mmol) in CH2Cl2(13 ml) was added EDCI (576 mg, to 3.09 mmol) and DMAP (366 mg, to 3.09 mmol) at 0°C. To the mixture was added dropwise a solution of acid 25 (1,11 g, 1.88 mmol) in CH2Cl2(rinsing with 5 ml + 2 ml) for 16 min at 0°C. After stirring at 0°C for 15 h, the mixture was stirred at RT for 3.5 hours. After concentration the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 30:1 to 20:1)to give 27 (1.20 g, of 1.61 mmol, 86%, based on the complex tert-butyl ether) as a colorless oil; [α]D24-25,1 (1,30, l3); IR (film) ν 2955, 2925, 2872, 1732, 1696, 1461, 1378, 1290, 1243, 1173, 1091, 985, 873, 773 cm-1;1H NMR (400 MHz, CDCl3) δ 0,06 (3H, c), 0,06 (3H, c), 0,58-0,66 (6H, m), 0,92 (N, c)of 0.95 (9H, t, J=8.0 Hz), of 1.02 (3H, d, J=6.5 Hz), of 1.03 (3H, d, J=6.5 Hz), with 1.07 (3H, c)to 1.21 (3H, c)to 1.67 (3H, c)2,07 (3H, c), 2,05-2,12 (1H, m), 2,30 (1H, DD, J=16,9, 7.5 Hz), 2,39 (1H, dt, J=14,8, 6,7 Hz), 2.49 USD (1H, DD, J=17,0, 3.0 Hz), 2,50 (1H, dt, J=14,8,6,7 Hz), 2,70 (3H, c), 2,74-of 2.30 (2H, m), of 3.07 (1H, DD, J=7,0 Hz), 3,83 (1H, DD, J=7,1, 2.0 Hz), of 4.35 (1H, DD, J=7,4, 2,8 Hz), 4,98 is 5.07 (4H, m), 5,16 (1H, ushort, J=7,0 Hz), 5,23 (1H, t, J=6.9 Hz), 5,74 (1H, DDT, J=16,7, to 10.2, 6.5 Hz), 5,91 (1H, DDD, J=17,8, 10,5, 7,8 Hz), 6,50 (1H, c), to 6.95 (1H, c);13With NMR (100 MHz, DCl3) was 3.7 δ, -3,3, 5,3 (3C), 7,2 (3C), 14,8, 15,2, 18,7, 18,9, 19,4, 20,3, 23,6, 23,7, 26,4 (3C), 31,7, 36,7, 40,1, 43,8,46,4, 53,3, 74,2, 76,5, 79,6, 115,5, 115,6, 116,5, 120,5, 121,3, 135,8, 136,1, 137,4, 140,2, 152,9, 164,7, 171,5, 218,4; MS-HP (ESI) calculated for C41H71NO5SSi2[M+Na+] 768,5 found 768,5.

Compound 39a:A solution of 27 (26.9 mg, 36,1 µmol) in toluene (70 ml) was boiled under reflux and treated with a solution of the catalyst of Grubbs (3.1 mg, 3.61 mmol) in toluene (2 ml). The mixture was stirred for 25 min, cooled to 0°C and filtered through a bed of silica gel, which was washed with hexane/EtOAc = 2/1. the joint the filtrate was concentrated and purified flash chromatography on a column (SiO 2, hexane/Et2O = 40:1 to 5:1), receiving 39a (9,9 mg of 13.8 mmol, 38%) as a colourless oil; [α]D25-41,5 (0,715, l3); IR (film) ν 2955, 2884, 1737, 1690, 1467, 1378, 1249, 1179, 1102, 1014, 979, 879, 826, 773 cm-1;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), 0,12 (3H, c), or 0.57 (6H, q, J=7,8 Hz)to 0.89 (9H, t, J=8.0 Hz), of 0.93 (9H, c), was 1.04 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), of 1.17 (3H, d, J=7,1 Hz), 1,68 (3H, c), of 2.15 (3H, d, J=0.8 Hz), 2,14-of 2.27 (2H, m), of 2.45 (1H, DD, J=14,0, 4,8 Hz), 2,50 (1H, DD, J=14,9, and 3.2 Hz), 2,64-to 2.74 (2H, m), of 2.72 (3H, c), to 3.02 (1H, Quint, J=7.0 Hz), 3,10 (1H, DD, J=14,4, 7,3 Hz), of 3.96 (1H, d, J=8.7 Hz), 4,43 (1H, DD, J=8,3, 2,9 Hz), with 5.22 (1H, DD, J=9,8, 5.7 Hz), 5,33-5,42 (2H, m), 5,69 (1H, DD, J=15,8, 8,2 Hz), to 6.57 (1H, c), of 6.96 (1H, c);13With NMR (100 MHz, DCl3) δ -3,3, -3,2, 5,6 (3C), and 7.1 (3C), 15,0, 17,2, 18,8, 19,4, 21,4, 21,7, 23,8, 24,3, 26,5 (3C), 33,2, 35,6, 41,3, 41,8, 48,2, 54,0, 74,4, 77,4, 79,3, 116,4, 120,5, 121,0, 129,3, 132,1, 137,8, 138,0, 152,7, 164,8, 170,7, 216,8; MS-HP (ESI) calculated for C39H68NO5SSi2[M+H+] 718,4 found 718,3.

Compound 28:To a solution of 39a (298,8 mg, 0,416 mmol) in THF (6.5 ml) was added HF-pyridine (3.2 ml) at 0°C and the mixture was stirred at RT for 3 hours. The reaction was suppressed by adding dropwise TMSOMe (30 ml) at 0°C, and the mixture was stirred at RT for 3 hours. After concentration and drying under high vacuum the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 1:1)to give 28 (196,6 mg, 0,402 mmol, 97%) as a white solid; [α]D25-96,6 (c 0,235, CHCl3); IR (film)ν 3502, 2970, 2927, 1733, 1685, 1506, 1456, 1375, 1251, 1152, 1040, 977 cm-1;1H NMR (400 MHz, CDCl3) δ of 1.06 (3H, c), is 1.11 (3H, d, J=7,0 Hz)to 1.22 (3H, d, J=6.8 Hz), 1.28 (in, 3H, c), 1,72 (3H, c), 2,10 (3H, c), 2,31-to 2.40 (2H, m), 2,43 (1H, DD, J=16,0, and 3.7 Hz), 2.49 USD (1H, DD, J=16,0, 9,2 Hz), 2,55 of 2.68 (2H, m), a 2.71 (3H, c), 2,98 (1H, DD, J=14,4, 6.4 Hz), and 3.16 (1H, Quint, J=6.2 Hz), 3,76 (1H, DD, J=5,9, and 3.2 Hz), 4,30 (1H, DD, J=9,2, and 3.7 Hz), 5,18 (1H, ushort, J=7,3 Hz), 5,32 (1H, DD, J=8,4, 2,5 Hz), 5,63 (1H, DD, J=15,7, 6.4 Hz), the ceiling of 5.60 (1H, DDD, J=15,7, 6,9, 5,1 Hz), 6,60 (1H, c), 6,98 (1H, c);13With NMR (100 MHz, DCl3) δ 15,1, 16,0, 17,7, 19,2, 19,5, 22,5, 23,6, 32,0, 35,0, 39,6, 40,3, 44,8, 53,3, 71,8, 75,6, 78,3, 116,1, 119,6, 120,5, 129,9, 131,3, 137,5, 138,2, 152,2, 165,0, 170,7, 218,8; MS-HP (ESI) calculated for C27H40NO5S [M+H+] 490,3 found 490,2.

Compound 26:Before the reaction the acid 25 and alcohol 21 was subjected to azeotropic distillation with dry benzene (5 ml × 2) and dried under high vacuum. To a solution of alcohol 21 (240 mg, 0,756 mmol) in CH2Cl2(5 ml) was added EDCI (192,7 mg, 1.01 mmol) and DMAP (122,8 mg, 1.01 mmol) at 0°C. To the mixture was added dropwise a solution of acid 25 (314,6 mg, 0,628 mmol) in CH2Cl2(washing 2 ml + 1 ml) for 15 min at 0°C. After stirring at 0°C for 2 h the mixture was stirred at RT for 2 hours. After concentration the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 20:1 to 15:1)to give 26 (340,1 mg, 0,425 mmol, 68% based on acid) as a colourless oil; [α]D24-27,5 (0,28, lsub> 3); IR (film) ν 2956, 2878, 1740, 1692, 1472, 1378, 1317, 1253, 1174, 1118, 988, 915, 872, 837, 775 cm-1;1H NMR (400 MHz, CDCl3) δ is 0.06 (6H, c), 0,57 is 0.65 (6H, m)to 0.92 (9H, c)to 0.94 (9H, t, J=7.9 Hz), of 1.02 (3H, d, J=6.9 Hz), of 1.03 (3H, d, J=6,8 Hz)of 1.07 (3H, c)to 1.22 (3H, c), 2,07-2,10 (1H, m), is 2.09 (3H, c), 2,31 (1H, DD, J=16.9 and, 7,3 Hz), of 2.51 (1H, DD, J=16,8, 3,0 Hz) 2,49-to 2.65 (2H, m), 2,71 (3H, c), 2,96-to 2.99 (2H, m), 3,06 (1H, Quint, J=7,1 Hz), 3,83 (1H, DD, J=7,3, and 2.1 Hz), 4,35 (1H, DD, J=7,2, 3.0 Hz), 4,98-5,12 (4H, m), and 5.30 (1H, t, J=of 6.7 Hz), USD 5.76 (1H, DDT, J=16,7,10,2, 6.2 Hz), of 5.92 (1H, DDD, J=17,8, 9,9, 7,8 Hz), to 6.19 (1H, t, J=7.0 Hz), 6,51 (1H, c), 6,97 (1H, c); MS-HP (ESI) calculated for C41H68F3NO5SSi2Na [M+Na+] 822,4 found 822,4.

Compound 40a (via RCM 26):A solution of 26 (57.6 mg, 72.0 mmol) in toluene (142 ml) was boiled under reflux and treated with a solution of the catalyst of Grubbs (6,1 mg, 7.20 mmol) in toluene (2 ml). The mixture was stirred for 28 min, cooled to 0°C and filtered through a bed of silica gel, which was washed with hexane/EtOAc = 2/1 (300 ml). The combined filtrates were concentrated and purified flash chromatography on a column (SiO2, hexane/Et2O = 40:1 to 15:2), receiving 40a (12.0 mg, of 15.5 mmol, 22%) as a colourless oil; IR (film) ν 2955, 2884, 1743, 1690, 1472, 1320, 1173, 1114,1038, 1008, 873, 832, 773 cm-1;1H NMR (400 MHz, CDCl3) δ 0,09 (3H, c), 0,12 (3H, c), of 0.55 (6H, q, J=7,7 Hz)to 0.88 (9H, t, J=8.0 Hz), is 0.96 (9H, c)a 1.01 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), of 1.20 (3H, d, J=7,1 Hz), 2,07-2,17 (1H, m), 2,19 (3H, c), of 2.38 (1H, DD, J=14,3, 3.5 Hz), ,39-2,49 (1H, m)of 2.50 (1H, DD, J=14,3, 7,3 Hz), 2,73 (3H, c), 2.77-to only 2.91 (2H, m), 2,96-to 3.09 (2H, m), 3,98 (1H, DD, J=8,9 Hz), of 4.54 (1H, DD, J=7,3, 3,4 Hz), 5,28 is 5.38 (1H, m), 5,63 (1H, DD, J=9,6, and 2.3 Hz), 5,77 (1H, DD, J=15,9 that 8.5 Hz), 6,21-6,28 (1H, m), 6,60 (1H, c), of 6.99 (1H, c); MS-HP (ESI) calculated for C39H65F3NO5SSi2[M+H+] 772,4 found 772,4.

Compound 29:To a solution of 40a (1.78 g, 2,31 mmol) in THF (25 ml) was slowly added HF·pyridine (12.5 ml) at 0°C and the mixture was stirred at RT for 4 hours. The reaction was suppressed by adding dropwise TMSOMe (80 ml) for 10 min at 0°C. the Mixture was vigorously stirred at RT for 2.5 hours. After concentration and drying under high vacuum for 2 h, the residue was purified flash chromatography on a column (SiO2~50 g, hexane/EtOAc = 1:1)to give 29 (1.20 g, 2.21 mmol, 96%) as a colourless powder; [α]D25-54,6 (0,28, l3); IR (film) ν 3478, 2974, 2929, 1736, 1689, 1449, 1381, 1318, 1247, 1169, 1113, 1039, 983, 867, 736 cm-1;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,8 Hz)to 1.37 (3H, c), 2,04 (1H, userd, J=3.8 Hz, HE), 2,12 (3H, c), 2,25 is 2.33 (1H, m), of 2.38 (1H, DD, J=15,3, 3.0 Hz), 2,48 (1H, DD, J=15,4, 9.8 Hz), 2,54-2,61 (1H, m), 2,66 was 2.76 (1H, m), 2,71 (3H, c), 2,96 (1H, DD, J=16,5,4,5 Hz), to 3.02 (1H, DD, J=16,3, 6.5 Hz), 3,11 (1H, Quint, J=6,7 Hz), 3,19 (1H, users, =OH), 3,74 (1H, users), 4,35 (1H, userd, J=9.5 Hz), 5,42 (1H, DD, J=6,2 by 4.1 Hz), the ceiling of 5.60 (1H, DDD, J=15,8, 5,6, and 4.5 Hz), to 5.66 (1H, DD, J=15,8, 5.8 Hz), 6,24 (1H, t, J=7.2 Hz), only 6.64 (1H, c), of 7.00 (1H, c);13C NMR (100 MHz (CDCl3) δ 15,1,16,1, 17,7, 18,5, 19,3, 22,5, 28,8 31,1, 39,6, 39,7, 45,0, 53,7, 71,4, 75,3, 76,8, 116,7, 120,2, 124,3 [kV1J(C,F) = 273,4 Hz], 127,9, 130,2 [kV3J(C,F) = 6,0 Hz], to 130.6 [kV2J(CF) = 28.4 Hz], 132,5, 136,7, 152,0, 165,4, 170,2, 218,4; MS-HP (ESI) calculated for C27H37F3NO5S [M+H+] 544,2 found 544,1.

Connection 2:To a solution of 29 (1,22 mg, 2,24 mmol) and NHNH2(26,7 mg, 89,6 mmol) in ClCH2CH2Cl (1 ml) at 50°C was added Et3N (12,5 µl, 89,6 Microm). The reaction was controlled by VETCH (hexane/EtOAc/CH2Cl2= 1/1/2). After stirring for 6.5 hours to the mixture was further added NHNH2(26,7 mg, 89,6 mmol) and Et3N (12,5 µl, 89,6 Microm). After stirring for 14 h the mixture was cooled to CT, diluted with EtOAc and filtered through a bed of silica gel, which is washed with EtOAc. After concentration the residue was purified preparative TLC (hexane/EtOAc/CH2Cl2=1/1/2), obtaining 2 (1,16 mg, 2,13 mmol, 94%) as white solids;1H NMR (400 MHz, CDCl3) δ of 1.03 (3H, d, J=7,0 Hz), a 1.08 (3H, c)to 1.19 (3H, d, J=6.8 Hz), 1,25-1,35 (2H, m)to 1.37 (3H, c), 1,42-of 1.55 (2H, m), 1,65-to 1.82 (2H, m), 2,10 (3H, d, J=0.8 Hz), 2.21 are 2,47 (2H, m), and 2.27 (1H, DD, J=14,2, 2,6 Hz), 2,48 (1H, DD, J=14,3, and 10.8 Hz), 2,70 (3H, c), 2,70-of 2.28 (1H, m), to 3.02 (1H, d, J=2.0 Hz, -OH), 3,19 (1H, CVD, J=6,9 and 2.2 Hz), the 3.65 (1H, d, J=6.2 Hz, -OH), 3,69-and 3.72 (1H, m), 4,34 (1H, DDD, J=10,8, 6,2, 2,6 Hz), 5,28 (1H, DD, J=10,2, 2.2 Hz), 6,12 (1H, DD, J=10,2, 5,2 Hz), is 6.61 (1H, c), 6,98 (1H, c); MS-HP (ESI) calculated for C27H39F3NO5S [M+H+]546,3, found 546,2.

The connection 54:Before the reaction the acid 25 and alcohol 53 was subjected to azeotropic distillation with dry benzene (3 ml × 2) and dried under high vacuum. To a solution of alcohol 53 (68,0 mg, 0,173 mmol) in CH2Cl2(1.3 ml) was added EDCI (of 37.8 mg, 0,197 mmol) and DMAP (24,1 mg, 0,197 mmol) at 0°C. To the mixture was added dropwise a solution of acid 25 (72,6 mg, 0,123 mmol) in CH2Cl2(0.7 ml) for 5 min at 0°C. After stirring at 0°C for 1 hour the mixture was stirred at RT for 2.5 hours. After concentration the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 30:1)to give 54 (99,5 mg, 0,114 mmol, 92%, based on the complex tert-butyl ether) as a colorless oil; [α]D25-23,4 (from 0.56, l3); IR (film) ν 2955, 2931, 2880, 1735, 1696, 1506, 1472, 1386, 1362, 1294, 1254, 1174, 1104, 988, 878, 776, 742 cm-1;1H NMR (400 MHz, CDCl3) δ 0,06 (3H, c), 0,06 (3H, c), of 0.14 (6H, c)to 0.63 (6H, q, J=8.0 Hz), to 0.92 (9H, c)to 0.94 (9H, t, J=8.0 Hz), of 0.97 (9H, c)of 1.02 (3H, d, J=6,6Hz), of 1.05 (3H, d, J=6.5 Hz), with 1.07 (3H, c)to 1.21 (3H, c)to 1.67 (3H, c), of 2.06 (3H, d, J=0.8 Hz), 2.05 is with 2.14 (1H, m), 2,30 (1H, DD, J=16,9, 7.5 Hz), 2,33 of $ 2.53 (2H, m), 2,50 (1H, DD, J=16.9 and, of 2.7 Hz), was 2.76 is 2.80 (2H, m)of 3.07 (1H, Quint, J=7.0 Hz), 3,83 (1H, DD, J=7,0, 2.2 Hz), 4,35 (1H, DD, J=7,4, 2,8 Hz), equal to 4.97 (2H, c), equal to 4.97 is 5.07 (4H, m), 5,16 (1H, t, J=7.2 Hz), of 5.24 (1H, t, J=6.9 Hz), 5,74 (1H, DDT, J=16,6, of 10.0, 6.5 Hz), 5,91 (1H, DDD, J=17,6, 9,9, 7,7 Hz), 6,50 (1H, c), 7,06 (1H, c);13C NMR (100 MHz (CDCl3) δ -5,2 (2C), was 3.7, -3,3, 5,3 (3C), 7,2 (3C), 14,7, 15,2, 18,5, 18,7, 18,9 20,3, 23,6, 23,7, 26,0 (3C)of 26.4 (3C), 31,7, 36,7, 40,1, 43,8, 46,4, 53,3, 63,4, 74,2, 76,5, 79,6, 115,5, 115,6, 116,6, 120,5, 121,3, 135,8, 136,1, 137,4, 140,1, 153,0, 171,5, 172,2, 218,4; MS-HP (ESI) calculated for C47H86NO6SSi3[M+H+] 876,6 found 876,5.

Compound 55:A solution of 54 (69,7 mg, 79.5 mmol) in toluene (158 ml) was boiled under reflux and treated with a solution of the catalyst of Grubbs (6,7 mg of 7.95 mmol) in toluene (2 ml). The mixture was stirred for 11 min, cooled to 0°C and filtered through a bed of silica gel, which was washed with hexane/EtOAc = 3/1 (280 ml). The combined filtrates were concentrated and purified flash chromatography on a column (SiO2, hexane/Et2O = from 20:1 to 15:1)to give 55 (18,4 mg, and 21.7 mmol, 27%) as a colourless oil; [α]D24-40,4 (from 0.26, l3); IR (film) ν 2955, 2930, 2879, 1740, 1694, 1472, 1387, 1362, 1253, 1200, 1107, 1007, 838, 776, 742 cm-1;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), 0,12 (3H, c), and 0.15 (6H, c), or 0.57 (6H, q, J=7.9 Hz), to 0.88 (9H, t, J=8.0 Hz), of 0.95 (9H, c)to 0.97 (9H, c), was 1.04 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), of 1.17 (3H, d, J=7,0 Hz), 1,69 (3H, c), 2.06 to 2,30 (2H, m), and 2.14 (3H, c), a 2.45 (1H, DD, J=15,6, 3.6 Hz), 2,50 (1H, DD, J=14,9, 3.1 Hz), 2,63 is 2.75 (2H, m), 2,97-of 3.06 (1H, m), 3,10 (1H, DD, J=14,6, 7,7 Hz), of 3.97 (1H, d, J=8.5 Hz), of 4.44 (1H, DD, J=8,4, 2,9 Hz), equal to 4.97 (2H, c), with 5.22 (1H, DD, J=8,7, 5,2 Hz), 5,33-5,44 (2H, m), 5,70 (1H, DD, J=15,6, 8.1 Hz), to 6.57 (1H, c), 7,07 (1H, c); MS-HP (ESI) calculated for C45H82NO6SSi3[M+H+] 848,5 found 848,5.

With the unity 57: To a solution of 55 (61,8 mg, for 72.8 mmol) in THF (2 ml) was added HF·pyridine (1 ml) at 0°C and the mixture was stirred at RT for 3.2 hours. The reaction was suppressed by adding dropwise TMSOMe (15 ml) at 0°C. the Mixture was stirred at RT for 2 hours. After concentration and drying under high vacuum the residue was purified flash chromatography on a column (SiO2, hexane/EtOAc = 1:3)to give 57 (32.4 mg, 64.1 mmol, 88%) as a white solid; [α]D25-108,4 (0,285, l3); IR (film) ν 3422, 2968, 2919, 2729, 1689, 1449, 1377, 1252, 1152, 1064, 978 cm-1;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=6,9 Hz)to 1.22 (3H, d, J=6.8 Hz), 1,32 (3H, c), 1,72 (3H, c), of 2.08 (3H, c), 2,31-to 2.40 (3H, m), 2,43 (1H, DD, J=15,5, 3.5 Hz), 2.49 USD (1H, DD, J=15,5, 9.5 Hz), 2,55-to 2.67 (2H, m), 2,95 (1H, DD, J=14,6, 6.3 Hz), 3,13 (1H, Quint, J=6.6 Hz), 3,34 (1H, users, HE's in), 3.75 (1H, DD, J=6,6, 2.4 Hz), 4,06 (1H, users, HE), to 4.33 (1H, DD, J=9,4, 3,0 Hz)to 4.92 (2H, c), is 5.18 (1H, t, J=6.9 Hz), 5,33 (1H, DD, J=8,0, 2,5 Hz), 5,52 (1H, DD, J=15,8, 6.4 Hz), 5,59 (1H, DDD, J=15,8, 6,6, 5.0 Hz), 6,63 (1H, c), 7,13 (1H, c);13C NMR (100 MHz, CDCl3) δ 15,3, 16,3, 17,8, 19,2, 22,8, 23,7, 31,9, 35,1, 39,7, 40,2, 45,0, 53,4, 61,8, 71,7, 75,8, 78,1, 116,7, 119,0, 120,5, 130,0, 131,2, 137,6, 138,9, 152,5, 170,0, 170,7, 218,7; MS-HP (ESI) calculated for C27H39NO6SNa [M+Na+] 528,2 found 528,0.

The connection 46:Before the reaction, the crude acid 25 (4,65 g, 7,27 mmol) and alcohol 44 (2,18 g, 9,84 mmol) was subjected to azeotropic distillation with dry benzene and dried under high vacuum. To a solution of JV the mouth 44 (2,18 g, 9,84 mmol) in CH2Cl2(65 ml) was added EDCI (2,09 g, 10.9 mmol) and DMAP (of 1.33 g, 10.9 mmol) at 0°C. To the mixture was added dropwise a solution of the crude acid 25 (4,65 g, 7,27 mmol) in CH2Cl2(rinsing with 20 ml + 5 ml) for 20 min at 0°C. After stirring at 0°C for 40 min, the mixture was stirred at RT for 4 hours. After concentration the residue was purified flash chromatography on a column (SiO2~160 g; hexane/EtOAc = 20:1)to give 46 (4,85 g, 6,87 mmol, 94% based on complex tert-butyl ether) as a colorless oil;1H NMR (400 MHz, CDCl3) δ 0,08 (3H, c), and 0.08 (3H, c), of 0.60 (6H, q, J=7,8 Hz)of 0.93 (9H, c)to 0.94 (9H, t, J=8.0 Hz), was 1.04 (3H, d, J=7,0 Hz), was 1.04 (3H, d, J=7,0 Hz), 1,11 (3H, c), of 1.23 (3H, c), 2,05 with 2.14 (1H, m), 2,17 (3H, c)that is 2.40 (1H, DD, J=16.9 and, 7,0 Hz), 2,59 (1H, DD, J=17,0, 3.6 Hz), 2,56-of 2.64 (2H, m), 2,90-a 3.01 (2H, m), 3,06 (1H, Quint, J=7.0 Hz), 3,85 (1H, DD, J=7,3, 2.0 Hz), to 4.38 (1H, d, J=7,0, 3,4 Hz), equal to 4.97-5,14 (5H, m), of 5.75 (1H, DDT, J=16,0, 9,9, 6.2 Hz), of 5.92 (1H, DDD, J=17,8, 10,5, 7,8 Hz), 6,21 (1H, TD, J=7,2, 1.5 Hz); MS-HP (ESI) calculated for C36H63F3O6Si2Na [M+Na+] 727,4 found 727,3.

The connection 48:A solution of 46 (510,0 mg, 0,723 mmol) in toluene (500 ml) was boiled under reflux and treated with a solution of the catalyst of Grubbs (of 92.1 mg, 0,109 mmol) in toluene (10 ml). The mixture was stirred for 17 min at boiling under reflux and immediately cooled to 0°C and kept at 0°C before being filtered through a pillow specification of the gel. The second party of diene (510,0 mg, 0,723 mmol) were processed identically and simultaneously. The combined reaction mixture was filtered through a bed of silica gel (100 g), which was washed with hexane/EtOAc = 3/1 (1.4 l). The combined filtrates were concentrated and purified flash chromatography on a column (SiO2~65 g, hexane/Et2O = from 10:1 to 5:1)to give 48 (742,4 mg, 1.10 mmol, 76%) as a colourless oil;1H NMR (400 MHz, CDCl3) δ 0,08 (3H), 0,10 (3H, c), of 0.60 (6H, q, J=7,8 Hz)of 0.93 (9H, c)to 0.94 (9H, t, J=7.8 Hz), of 1.03 (3H, d, J=7,1 Hz), a 1.08 (3H, c), of 1.13 (3H, d, J=7,0 Hz)of 1.17 (3H, c), and 2.26 (3H, c), 2,25-of 2.34 (1H, m), 2,64 (1H, DD, J=15,5, 5.0 Hz), 2,68 is 2.75 (2H, m), was 2.76 (1H, DD, J=15,6, 6.4 Hz), 2,85 (1H, DD, J=15,6, 5.7 Hz), 2,97 (1H, CVD, J=8,3, 6.9 Hz), totaling 3.04 (1H, DD, J=15,6, 6.3 Hz), 3,92 (1H, DD, J=8,3, 1.2 Hz), 4,36 (1H, t, J=at 5.3 Hz), 5,30 of 5.39 (2H, m), to 5.58 (1H, DD, J=15,5, 8.0 Hz), 6,13 (1H, ushort, J=7.2 Hz);13C NMR (100 MHz, CDCl3) δ -3,6, -3,6, 5,4 (3C), 7,0 (3C), 17,5, 18,5, 19,0, 21,6, 23,5, 26,3 (3C), 26,5, 28,6, 29,1, 41,0, 42,3, 47,3, 54,1, 74,2, 76,8, 77,7, 124,0 [1J(C,F)=273,7 Hz], 126,0, 128,7 [3J(C,F)=5,9 Hz], 132,2 [2J(C,F)=28,1 Hz], 133,8, 170,5, 204,1, 216,1; MS-HP (ESI) calculated for C34H59F3O6Si2Na [M+Na+] 699,4 found 699,4.

Compound 40a (via Wittig reaction of the ketone 48):Ketone 48 was subjected to azeotropic distillation with benzene (5 ml × 2) and then dried under high vacuum for 0.5 hour. To a solution of Wittig salt (907 mg, at 2.59 mmol) in THF (19 ml) was added dropwise t-BuOK (2.4 ml 1.0 M RA is down in THF, 2,43 mmol) over 5 min at 0°C. the Mixture was stirred at 0°C for 0.5 hour and then was cooled to

-78°C. To the mixture was added dropwise a solution of ketone 48 (1.10 g, of 1.62 mmol) in THF (13 ml) for 10 min and the resulting mixture was allowed to warm to -20°C for 2 hours. The reaction extinguished saturated aqueous NH4Cl (15 ml) and was extracted with EtOAc (50 ml × 3). The combined organic layers were washed with saturated salt solution (20 ml), dried over Na2SO4and concentrated. The residue was purified flash chromatography (SiO2, hexane/Et2O = from 20:1 to 10:1), obtaining the required 16(E)-isomer 40a (940 mg, 1,22 mmol, 75%) together with unwanted 16(Z)-isomer 40b (140,9 mg of 0.182 mmol, 11%), both as colourless oils;

[α]D26-17,1 (0,14, l3);1H NMR (400 MHz, CDCl3) δ 0,09 (3H, c), 0,12 (3H, c), of 0.55 (6H, q, J=7,7 Hz)to 0.88 (9H, t, J=8.0 Hz), is 0.96 (9H, c)a 1.01 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), of 1.20 (3H, d, J=7,1 Hz), 2,07-2,17 (1H, m), 2,19 (3H, c), of 2.38 (1H, DD, J=14,3, 3.5 Hz), 2,39-2,49 (1H, m), 2,50 (1H, DD, J=14,3, 7,3 Hz), 2,73 (3H, c), 2.77-to only 2.91 (2H, m), 2,96-to 3.09 (2H, m), 3,98 (1H, DD, J=8,9 Hz), of 4.54 (1H, DD, J=7,3, 3,4 Hz), 5,28 is 5.38 (1H, m), 5,63 (1H, DD, J=9,6, and 2.3 Hz), 5,77 (1H, DD, J=15,9, 8.5 Hz), 6,21-6,28 (1H, m), 6,60 (1H, c), of 6.99 (1H, c);13C NMR (100 MHz, CDCl3) δ -3,4,-3,3, 5,5 (3C), 7,0 (3C), 14,6, 17,1, 18,7, 19,4, 19,9, 21,3, 24,8, 26,4 (3C), 29,6, 32,8, 42,0, 42,1, 48,2, 54,1, 73,4, 76,9, 77,8, 117,0, 121,6, 124,3 [1J(C,F) = 273,5 Hz], 127,2, to 130.6 [2J(C,F) = 28,2 Hz], 130, 8mm [3J(C,F) = 6,1 Hz], 133,2, 136,5, 152,3, 165,0,170,1, 217,1; MS-HP (ESI) calculated for C39H65F3NO5SSi2[M+H+] 772,4074 found 772,4102.

[α]D2562,7 (from 0.33, l3);1H NMR (400 MHz, CDCl3) δ 0,09 (3H, c), 0,13 (3H, c), 0,49 (6H, q, J=7,8 Hz)to 0.85 (9H, t, J=7,8 Hz)to 0.97 (9H, c)0,99 (3H, c), of 1.06 (3H, d, J=7,1 Hz), 1,11 (3H, c), of 1.20 (3H, d, J=7,1 Hz), from 2.00 (3H, c), 2,03 and 2.13 (1H, m), 2,35 (1H, DD, J=14,3, 3.0 Hz), 2,46 (1H, DD, J=14,3, and 7.8 Hz), 2,41-of 2.50 (1H, m), 2,73 (3H, c), 2,71-2,90 (2H, m) 2,98-of 3.12 (2H, m), 3,99 (1H, d, J=9,2 Hz), 4,56 (1H, DD, J=7,7, 2,8 Hz), 5,33 (1H, DDD, J=15,6, 8,9, the 4.1 Hz), of 5.82 (1H, DD, J=15,6, and 8.4 Hz), of 6.29 (1H, c), 6,33-6,40 (1H, m)6,94 (1H, m), to 7.09 (1H, userd, J=8,4 Hz);13C NMR (100 MHz, CDCl3) δ -3,2, -3,2, 5,5 (3C), 7,0 (3C), 17,2, 18,7, 19,3, 19,6, 20,0, 22,3, 24,9, 26,4 (3C), 29,7, 32,9, 41,9, 42,0, 48,6, 54,0, 72,2, 73,3, 77,0, 116,7, 120,7, 124,5 [1J(C,F) = 273,3 Hz], 127,9, 129,7 [2J (C,F) = 28,0 Hz], 131,9 [3J (C,F) = 6,1 Hz], 132,9, 136,6, 152,1, 165,4, 170,2, 217,4; MS-HP (ESI) calculated for C39H65F3NO5SSi2[M+H+] 772,4 found 772,4.

Compound 58 (via Wittig reaction of the ketone 48):Ketone 48 was subjected to azeotropic distillation with benzene (5 ml × 2) and then dried under high vacuum for 0.5 hour. To a solution of Wittig salt (1.19 g, and 2.27 mmol) in THF (18 ml) was added dropwise t-BuOK (2.2 ml of 1.0 M solution in THF, of 2.20 mmol) over 5 min at 0°C. the Mixture was stirred at 0°C for 20 min and then was cooled to

-78°C. To the mixture was added dropwise a solution of ketone (1.06 g, is 1.51 mmol who) in THF (rinsing with 10 ml + 2 ml) for 10 min and the resulting mixture was allowed to warm to -20°C for 2 hours. The reaction extinguished saturated aqueous NH4Cl (15 ml) and was extracted with EtOAc (50 ml × 3). The combined organic layers were washed with saturated salt solution (20 ml), dried over Na2SO4and concentrated. The residue was purified flash chromatography on a column (SiO2~65 g, hexane/Et2O = 30:1 to 20:1), obtaining the required 16(E)-isomer 58 (1.01 g, 1.11 mmol, 74%) together with unwanted 16(Z)-isomer 58a (154,5 mg of 0.182 mmol, 11%), both as colourless oils;

1H NMR (400 MHz, CDCl3) δ 0,09 (3H, c), 0,12 (3H, c), and 0.15 (6H, c)of 0.55 (6H, q, J=7,8 Hz)of 0.87 (9H, t, J=8.0 Hz), is 0.96 (9H, c)to 0.97 (9H, c)a 1.01 (3H, c), of 1.06 (3H, d, J=7,1 Hz)of 1.12 (3H, c), of 1.20 (3H, d, J=7,1 Hz), 2,07-of 2.16 (1H, m)to 2.18 (3H, d, J=1.0 Hz), of 2.38 (1H, DD, J=14,4, and 3.3 Hz), 2,34 is 2.46 (1H, m), 2.49 USD (1H, DD, J=14,4, 7,4 Hz), 2,78-2,90 (2H, m), 2,97-to 3.09 (2H, m), 3,98 (1H, d, J=8,9 Hz), of 4.54 (1H, DD, J=7,3, 3,3 Hz), equal to 4.97 (2H, c), 5,33 (1H, DDD, J=15,8, 8,6, a 4.9 Hz), 5,63 (1H, DD, J=a 9.6, 2.4 Hz), 5,78 (1H, DD, J=15,8, 8,2 Hz), 6.22 per 6,27 (1H, m), 6,60 (1H, c), to 7.09 (1H, c);13C NMR (100 MHz, CDCl3) δ -5,3 (2C), -3,4, -3,3, 5,5 (3C), 7,0 (3C), 14,6, 17,1, 18,4, 18,7, 19,8, 21,3, 24,8, 25,9 (3C)of 26.4 (3C), 29,6, 32,9, 42,0, 42,1, 48,2, 54,1, 63,4, 73,4, 76,9, 77,8, 117,2, 121,7, 124,3 [kV1J(C,F)=273,6 Hz], 127,2, 130,7 [kV2J(C,F) = 27,5 Hz], 130, 8mm [kV3J(C,F) = 6.2 Hz], 133,2, 136,4, 152,6, 170,1, 172,4, 217,1; MS-HP (ESI) calculated for C45H78F3NO6SSi3Na[M+Na+] 924,5 found 924,5.

1H NMR (400 MHz, CDCl3) δ 0,07 (3H, c), 0,13 (3H, c), 0,16 (6H, c)0,48 (6H, q, J=7.8 Hz), is 0.84 (9H, t, J=7.9 Hz), 0,97 (18H, c), and 0.98 (3H, c), of 1.06 (3H, the, J=7,1 Hz), 1,11 (3H, c), of 1.20 (3H, d, J=7,2 Hz), from 2.00 (3H, c), 2,03-2,11 (1H, m), 2,33 (1H, DD, J=14,1, 2,8 Hz), 2,43 (1H, DD, J=14,0, and 7.8 Hz), 2.40 a-2,48 (1H, m), was 2.76-2,89 (2H, m), 2,97-3,10 (2H, m), 3,99 (1H, d, J=9.3 Hz), of 4.57 (1H, DD, J=7,8, and 2.6 Hz), of 4.95 (1H, d, J=14.6 Hz), 5,00 (1H, d, J=14.6 Hz), 5,33 (1H, DDD, J=15,6, 9,1, 3.8 Hz), of 5.82 (1H, DD, J=15,6, 8,3 Hz), 6,30 (1H, c), 6,32-6,38 (1H, m),? 7.04 baby mortality (1H, c), 7,11 (1H, DD, J=11,0, 2,3 Hz); MS-HP (ESI) calculated for C45H78F3NO6SNa[M+Na+] 924,5 found 924,5.

Compound 59:To a solution of 58 (1.04 g, 2.25 mmol) in THF (22 ml) was slowly added HF·pyridine (11 ml) at 0°C and the mixture was stirred at RT for 4.3 hours. The reaction was suppressed by adding dropwise TMSOMe (75 ml) for 10 min at 0°C. the Mixture was vigorously stirred at RT for 4.2 hours. After concentration and drying under high vacuum for 1 hour, the residue was purified flash chromatography on a column (SiO2~25 g; hexane/EtOAc = 3:4 to 1:2)to give 59 (615,7 mg, 1.00 mmol, 96%) as a colourless powder; [α]D25-57,7 (1,20, l3);1H NMR (400 MHz, CDCl3) δ 1.04 million (3H, c), of 1.12 (3H, d, J=6.9 Hz), 1,25 (3H, d, J=6.8 Hz), of 1.36 (3H, c), 1,90 (1H, d, J=6,6 Hz, OH), of 2.08 (3H, c), 2,23 of-2.32 (1H, m), 2,34 (1H, DD, J=15,7, 2.4 Hz), 2.49 USD (1H, DD, J=15,7, 10.1 Hz), 2,59-2,69 (2H, m), 2.95 and-a 3.01 (2H, m), 3.04 from (1H, quintet, J=6.8 Hz), and 3.72 (1H, dt, J=7,0, 3.0 Hz), of 3.78 (1H, d, J=5.7 Hz, HE), to 4.38 (1H, DDD, J=10,1, 5,7, 2.4 Hz), the 4.90 (2H, d, J=6,1 Hz), 5,10 (1H, t, J=6,1 Hz, HE), 5,44 (1H, t, J=4,7 Hz), the ceiling of 5.60 (1H, DD, J=15,9, 4,4 Hz), to 5.66 (1H, DD, J=15,9, 5.0 Hz), 6,28 (1H, t, J=6,7 Hz), was 6.73 (1H, c), 7,16 (1H, c); MS-HP (ESI) calculated for sub> 27H37F3NO6SNa[M+H+] 560,2 found 560,1.

Compounds 49 and 50:A solution of 28 (12,2 mg, 24,9 mmol) in CH2Cl2(1.25 ml) was cooled to -78°C and treated with a cooled solution of DMDO (-78°C 0,06 M in acetone; 914 μl, of 54.8 mmol). The mixture was allowed to warm to -50°C and stirred at -50°C for 2.7 hours. Excess DMDO extinguished at -50°C the addition of dimethyl sulfide (117 μl) and the mixture was stirred at this temperature for 0.5 hour. The solvent was removed in vacuum. Purification preparative thin-layer chromatography (hexane/EtOAc = 1/2) gave β-epoxide 49 (3.0 mg, to 5.93 mmol, 24%) and α-epoxide 50 (7.9 mg, 15.6 mmol, 63%), both as colourless solid.

The connection 49:1H NMR (400 MHz, CDCl3) δ of 1.03 (3H, s), is 1.11 (3H, d, J=7,0 Hz)to 1.14 (3H, d, J=6,9 Hz)of 1.34 (3H, c), of 1.36 (3H, c), from 2.00 (1H, DDD, J=15,1, 7,3, 4.0 Hz), and 2.14 (1H, dt, J=15,1, 5,2 Hz), and 2.14 (3H, c), of 2.21 (1H, DD, J=14,6, 8.0 Hz), 2,33 (1H, DD, J=14,7, 4,8 Hz), 2,47 (1H, DD, J=13,8, and 3.3 Hz), 2,59 (1H, DD, J=13,8, and 9.4 Hz), 2,73 (3H, c), 2,77 (1H, users, HE), with 2.93 (1H, DD, J=7,3, 4,8 Hz)to 3.34 (1H, CVD, J=6,9, and 3.7 Hz), 3.75 to 3,82 (1H, m), 4,12-4,24 (2H, m,including HE), 5,54 (1H, DDD, J=15,7, 7,4, 5.0 Hz), 5,54-the ceiling of 5.60 (1H, m), 5,64 (1H, DD, J=15,7, 5.6 Hz), 6,94 (1H, c), 7,01 (1H, c); MS-HP (ESI) calculated for C27H40NO6S[M+N+] 506,3 found 506,3.

Compound 50:1H NMR (400 MHz, CDCl3) δ and 1.00 (3H, c), was 1.04 (3H, d, J=6.9 Hz), 112 (3H, d, J=7,0 Hz)of 1.35 (3H, c), of 1.35 (3H, c)to 1.87 (1H, dt, J=15,0, 9,2 Hz), 2,03 (1H, DD, J=13,9, 9,2 Hz), 2,13 (3H, c), 2,13-2,19 (1H, m), a 2.36 (1H, DD, J=13,9, 3,4 Hz), 2,39 (1H, DD, J=12,2, 2,1 Hz), 2,42 is 2.51 (1H, m), 2.49 USD (1H, DD, J=12,4, up 10.9 Hz), 2,69 (1H, d, J=2.7 Hz), of 2.72 (3H, c), 3,06 (1H, DD, J=9,7, 3.1 Hz), 3,54 (1H, CVD, J=7,0, 2.0 Hz), 3,76-of 3.80 (1H, m), 4,07-to 4.14 (1H, m), or 4.31 (1H, d, J=4,1 Hz), 5,52 (1H, DD, J=15,5, and 8.7 Hz), the ceiling of 5.60 (1H, DDD, J=15,1, 9,4, 3,4 Hz), 5,71 (1H, d, J=8,4 Hz), 6,63 (1H, c), of 6.99 (1H, c); MS-HP (ESI) calculated for C27H39NO6SNa[M+Na+] 528,2 found 528,2.

The connection 52:To a solution of 50 (1.7 mg, 3.4 mmol) and NHNH2(40,1 mg, 0,134 mmol) in ClCH2CH2Cl (0.8 ml) at 50°C was added Et3N (18,7 μl, 0,134 mmol). The reaction was controlled by VETCH (hexane/EtOAc = 1/2). After stirring for 4 h the mixture was cooled to CT, diluted with EtOAc and filtered through a bed of silica gel, which is washed with EtOAc. After concentration the residue was purified preparative TLC (hexane/EtOAc = 1/2)to give 52 (1.2 mg, 2.4 mmol, 70%) as a white solid.1H NMR (400 MHz, CDCl3) δ of 0.95 (3H, d, J=7,1 Hz), was 1.04 (3H, c), is 1.11 (3H, d, J=7.0 Hz), 1.28 (in, 3H, c), of 1.37 (3H, c), 1,35-of 1.44 (1H, m), 1,45-to 1.59 (4H, m), 1,71-to 1.82 (2H, m)to 1.86 (1H, dt, J=15,3, 9.5 Hz), 2,10 (1H, DD, J=15,3, 3,6 Hz)by 2.13 (3H, c), is 2.40 (1H, DD, J=12,5, 2,5 Hz), 2.49 USD (1H, DD, J=12,5, and 11.0 Hz), is 2.74 (3H, c), 2,80 (1H, users, HE), of 3.07 (1H, DD, J=10,3, 3,3 Hz)to 3.34 (1H, CVD, J=7,0, 1.0 Hz), the 3.89 (1H, users, HE), a 4.03-4.09 to (1H, m), 4,12-4,17 (1H, m), 5,69 (1H, d, J=9.1 Hz), 6,63 (1H, c), of 7.00 (1H, c); MS-HP (ESI) calculated for C27H41NO6SNa[M+Na+us $ 530, 3, found 530,2.

The connection 51:To a solution of 49 (0.7 mg, 1.38 mmol) and NHNH2(20.6 mg, 69 μmol) in ClCH2CH2Cl (0.4 ml) at 50°C was added Et3N (a 9.6 μl, 69 mmol). The reaction was controlled by VETCH (hexane/EtOAc = 1/2). After stirring for 6 h the mixture was cooled to CT, diluted with EtOAc and filtered through a bed of silica gel, which is washed with EtOAc. After concentration the residue was purified preparative TLC (hexane/EtOAc = 1/2)to give 51 (0.5 mg, 0,985 mmol, 71%) as a white solid. Spectral data 51 were identical to the data described for EpoB.

Example 2

Alternative methods of synthesis for the synthesis of intermediate products epothilone

The following examples are offered ways to obtain various intermediate products of the synthesis of analogues epothilones.

Optimization of the synthesis of 9,10-dehydroemetine

Example 1

Example 2

Recovery Noyori

Example 3

Recovery Noyori

Example 4

Alternative synthesis of key diketone

Example 5

Method 1. Migration of the silyl group - decarboxylation

Method 2. Decarboxylation is the introduction of a silyl group

Example 6

An additional method of synthesis of 2-hydroxyketone Evans

Example 7

The method of synthesis of 2-hydroxyketone Kowalski-he sang sharpless

Experiments

1-(2-Benzyloxy-1-methylethyl)-5,5-diisopropoxide-2,4,4-trimethyl-3-oxopentanoic; 2,2,2-trichlorethylene fluids carboxylic acid (32a)

To a solution of 7-benzyloxy-5-hydroxy-1,1-diisopropoxide-2,2,4,6-tetramethylheptane-3-it 32 (1.0 g, 2.4 mmol) and pyridine (0.8 ml, 7,3 mmol) in CH2Cl2(10.0 ml) at 0°C was added 2,2,2-trihloretilamina (668,0 μl, 4.9 mmol) and the mixture was allowed to warm up to CT. After 1 hour the reaction mixture is extinguished saturated salt solution and then was extracted with CH2Cl2. The combined organic layers were dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 93:7), receiving 32a (1,285 g, 92%) as a clear oil:1H NMR (400 MHz, CDCl3) δ 1,03-of 1.09 (m, 12H)and 1.15 (d, J=1.8 Hz, 3H), of 1.17 (d, J=1.9 Hz, 3H), 1,19-to 1.21 (m, 6H), 1,97-2,11 (m, 1H), 3,2 (DD, J=6.2 and 9.0 Hz, 1H), 3,54 (DD, J=4.8 and 9.1 Hz, 1H), 3,57-of 3.60 (m, 1H), 3,82 (CVD, J=3,6 and 5.9 Hz, 2H), 4,47 (c, 2H), 4,57 (c, 1H), 4.72 in (d, J=11,9 Hz, 1H), 4,81 (d, J=11,9 Hz, 1H), 5,08 (t, J= 6.0 Hz, 1H), 7,29-to 7.35 (m, 5H);13With NMR (100 MHz, DCl3) δ 11,9, 15,0, 18,8, 21,4, 21,7, 22,3, 23,2, 23,4, 35,7, 42,5, 53,4, 53,9, 69,4, 70,9, 71,4, 73,3, 81,3, 94,7, 103,4, 127,5, 127,6, 128,2, 138,2, 154,0, 215,6; IR (film, NaCl, cm-1) 2966, 1760, 1698, 1247; MS-HP (ESI) calculated for C27H41About7Cl3Na [M+Na+] 605,2 found 605,2; [α]23D= -20,4 (c = 1,0, CHCl3).

1-(2-Benzyloxy-1-methylethyl)-2,4,4-trimethyl-3,5-dioxopentanoate; 2,2,2-trichlorethylene fluids carboxylic acid (67)

To a solution of 32a (1.28 g, 2.25 mmol) in 4:1 THF/H2O (25 ml) was added p-TsOH (111,0 mg, 0.6 mmol). After heating at 70°C for 5 h, the reaction mixture was poured into cold (0°C) saturated aqueous solution of NaHCO3(12 ml) and then was extracted with EtOAc. The combined organic layers were washed with saturated salt solution, dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 84:16), receiving 67 (793,2 mg, 76%) as a clear oil:1H NMR (400 MHz, CDCl3) δ of 0.90 (d, J=5.8 Hz, 3H), 1,0 (d, J=6.9 Hz, 3H), 1,24 (c, 6H), 1,97-2,04 (m, 1H), 3,24 (DD, J=4,8 and 9.2 Hz, 1H), 3,34 (m, 1H), 3,42 (DD, J=5,8 and 9.2 Hz, 1H), 4,35 (d, J=11,9 Hz, 1H), 4,39 (d, J=11,9 Hz, 1H), with 4.64 (d, J=11,9 Hz, 1H), 4,69 (d, J=11,9 Hz, 1H), 4,96 (t, J=6.0 Hz, 1H), 7,19-7,28 (m, 5H), 9,49 (c, 1H);13With NMR (100 MHz, DCl3) -12,0, 14,8, 19,5, 19,6, 35,4, 43,3, 60,9, 71,1, 73,3, 80,37, 94,5, 127,7, 127,8, 128,3, 137,9, 154,1, 201,0, 210,1; IR (film, NaCl, cm-1) 2973, 2880, 1758, 1701, 1453 1380, 1248; MS-HP (ESI) calculated for C21H27About6Cl3Na [M+Na+] 503,0 found 503,0; [α]23D= is 18.5 (c = 0.8, the CHCl3).

Tert-butyl ester 9-benzyloxy-4,4,6,8-tetramethyl-3,5-dioxo-7-(2,2,2-trichlorocarbanilide)nonanalog acid (69)

To a solution of LDA (1,17 mmol, 0.3 M in Et2O) at -78°C was added tert-butyl acetate (1.0 mmol, 135,0 µl). After 30 min slowly over 15 min was added to a solution of 67 (464,0 mg, 1 mmol) in Et2O (2 ml). After stirring for 1 hour the reaction was suppressed saturated aqueous NH4Cl and then was extracted with EtOAc. The combined organic layers were washed with saturated salt solution, dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 86:14), receiving 68 (1:1 mixture of epimeres, 461,4 mg, 80%) as a clear oil:1H NMR (400 MHz, CDCl3) δ of 0.87 (d, J=5.3 Hz, 3H), 0,89 (d, J=5.5 Hz, 3H), 1,02-of 1.10 (m, 18H), 1,38 (c, 18H), 1,97-2,2 (m, 2H), 2,27-2,31 (m, 2H), 3,22-of 3.27 (m, 3H), 3,39-of 3.48 (m, 5H), a 4.03-4,06 (m, 1H), 4,11-to 4.14 (m, 1H), of 4.38 is 4.45 (m, 4H), 4,58-to 4.73 (m, 4H), equal to 4.97 (t, J=5.8 Hz, 1H), 5,02 (t, J=5.8 Hz, 1H), 7.18 in-7,27 (m, 10H);13With NMR (100 MHz, DCl3) δ 11,9, 12,7, 14,9, 15,2, 18,7, 19,3, 21,4, 21,6, 28,0, 35,6, 37,4, 41,7, 42,0, 51,8, 51,9, 71,3, 71,3, 72,5, 73,0, 73,3, 73,3, 80,6, 81,2, 81,3, 94,6, 127,5, 127,7, 127,8, 128,3, 138,0, 138,1, 154,0, 154,1, 172,3, 172,4, 216,0, 216,3; IR (film, NaCl, cm-1) 3509, 2975, 1759, 1707, 1368, 1248, 1152; MS-HP (ESI) bycicle what about for 27H39About8Cl3Na [M+Na+] 619,1 found 619,2.

To a solution of 68 at 0°C (350,0 mg, 0.6 mmol) in CH2Cl2(10 ml) was added periodinane dess-Martin (398,0 mg, 0.9 mmol). The mixture was stirred at RT for 1 hour and then was poured into a well stirred mixture of 1:1 saturated Na2S2O3/saturated NaHCO3. The layers were separated after 30 minutes the Aqueous layer three times was extracted with Et2O. the combined organic extracts were washed with saturated NaHCO3saturated salt solution, dried over MgSO4and concentrated in vacuum. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 91:9)to give 69 (258,4 mg, 74%) as a clear oil:1H NMR (400 MHz, CDCl3) δ 0,80 (d, J=6.9 Hz, 3H), of 0.87 (d, J=6.9 Hz, 3H), 1,13 (c, 3H), 1,19 (c, 3H), 1,23 (c, 9H), 2,04-2,12 (m, 1H), 3,09 of 3.28 (m, 5H), 4,23 (c, 2H), 4,48(d, J=11,9 Hz, 1H), 4,55 (d, J=11,9 Hz, 1H), 4,79 (DD, J=4,6 and 7.3 Hz, 1H),? 7.04 baby mortality-7,13 (m, 5H);13With NMR (100 MHz, DCl3) δ 11,7, 14,6, 20,7, 21,5, 27,9, 35,5, 42,2, 43,4, 63,3, 71,3, 73,3, 79,9, 81,5, 90,5, 94,5, 127,6, 127,7, 128,2, 138,0, 154,0, 166,2, 202,9, 210,0; IR (film, NaCl, cm-1) 2977, 1758, 1697, 1368, 1248, 1154; MS-HP (ESI) calculated for C27H37About8Cl3Na [M+Na+] 617,1 found 617,1; [α]23D= -49,1 (c = 0,9, CHCl3).

Tert-butyl ester 9-benzyloxy-3-hydroxy-4,4,6,8-tetramethyl-5-oxo-7-(2,2,2-trichlorocarbanilide)nonanalog acid (70)

GI is ISO high pressure loaded catalyst (R)-RuBINAP (16,8 mg, 10.0 µmol). Was added HCl (555 μl, 0,2N in MeOH) and then the mixture was treated with ultrasound for 15 sec. Then was added a solution of 69 (59,4 mg, 0.1 mmol) in MeOH (555 μl) and the mixture was transferred into a Parr apparatus. The vessel was purged H2within 5 min, and then created a pressure up to 1200 psi. After 17 h the reaction mixture was returned to conditions of atmospheric pressure and poured into a saturated aqueous solution of NaHCO3. The aqueous layer three times was extracted with EtOAc. The combined organic extracts were dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 88:12), receiving 70 (dr>20:1 based on the1H-NMR analysis), or 47.6 mg, 80%) as a colorless oil:1H NMR (400 MHz, CDCl3) δ of 1.06 (d, J=6.9 Hz, 3H), 1,11 (d, J=6.8 Hz, 3H), 1.14 in (c, 3H), 1.18 to (c, 3H), 1,47 (c, 9H), 2.05 is-a 2.12 (m, 1H), 2,35-to 2.40 (m, 1H), 3,31-3,37 (m, 2H), 3,51-of 3.54 (m, 2H), 4,11-to 4.14 (m, 1H), 4,46 (c, 2H), 4.72 in (d, J=11,9 Hz, 1H), 4,80 (d, J=11,9 Hz, 1H), of 5.05 (DD, J=5,0 and 6.7 Hz, 1H), 7,27-to 7.35 (m, 5H);13With NMR (100 MHz, DCl3) δ 12,0, 15,0, 19,3, 21,7, 28,0, 35,6, 37,5, 41,7, 51,8, 71,3, 73,0, 73,3, 80,6, 81,3, 94,7, 127,5, 127,7, 128,3, 138,2, 154,1, 172,4, 216; IR (film, NaCl, cm-1) 3849, 2974, 2879, 1758, 1701, 1454, 1368, 1248, 1152, 926, 734; MS-HP (ESI) calculated for C27H39About8Cl3Na [M+Na+] 619,1 found 619,2; [α]23D= -13,0 (c = 0.4, CHCl3).

Tert-butyl ester 9-benzyloxy-4,4,6,8-tetramethyl-5-OK the-7-(2,2,2-trichlorocarbanilide)-3-(triethylsilyl)nonanalog acid (71)

To a solution of 70 (37.6 mg, 6.3 mmol) and imidazole (9.4 mg, of 13.8 mmol) in DMF (0.4 ml) at 0°C was added TESCl (11,6 ál and 69.3 mmol). After 3 h the mixture was diluted with saturated aqueous NaHCO3. The aqueous layer three times were extracted with hexane. The combined organic extracts were washed with saturated salt solution, dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 93:7)to give in order of elution 71 (22.9 mg, 51%) and extracted 70 (12.9 mg, 34%) as colourless oils. 7:1H NMR (400 MHz, CDCl3) δ 0,66 (kV, J=7.9 Hz, 6H), is 0.96 (t, J=7.9 Hz, 9H), 1,01 (c, 3H), of 1.05 (d, J=5,2 Hz, 3H), of 1.07 (d, J=5.3 Hz, 3H), 1,35 (c, 3H), 1,44 (c, 9H), 2.05 is-2,11 (m, 2H), 2,50 (DD, J=3,5 and 17.2 Hz, 1H), 3,35 (DD, J=of 5.9 and 9.0 Hz, 1H), 3,49 (DD, J=4.0 and 9.0 Hz, 1H), 3,53 (DD, J=3,8 and 6.7 Hz, 1H), 4,18 (DD, J=3.5 and 6.5 Hz, 1H), 4,45 (c, 2H)and 4.65 (d, J=11,9 Hz, 1H), 4,79 (d, J=11,9 Hz, 1H), equal to 4.97 (DD, J=3.7 and 8.1 Hz, 1H), 7,29-7,52 (m, 5H);13With NMR (125 MHz, DCl3) δ 5,3, 7,3, 10,9, 14,9, 21,3, 22,6, 28,4, 35,9, 41,1, 42,7, 53,7, 71,9, 73,7, 75,7, 80,1, 80,9, 95,1, 127,9, 128,0, 128,7, 138,6, 154,3, 171,7, 215,7; IR (film, NaCl, cm-1) 2956, 2876, 1732, 1694, 1456, 1366, 1257, 1154, 1098, 988, 835, 774, 741; MS-HP (ESI) calculated for C33H53About8SiCl3Na [M+Na+] 733,2 found 733,3; [α]23D= -16,1 (c = 0,1, CHCl3).

Tert-butyl ester 9-benzyloxy-3-(diethylethylenediamine)-7-hydroxy-4,4,6,8-tetramethyl-5-oksanalove acid (71a)

It races the thief 71 (22.9 mg, 3.2 µmol) in a mixture of 1:1 THF/AcOH (1,4 ml) was added Zn (5.0 mg, 7.8 mmol, nano-sized). The mixture was treated with ultrasound for 15 minutes was Added Zn (5.0 mg, 7.8 mmol, nano-sized), and then treated with ultrasound for 15 minutes the Suspension was filtered through a pad celite, rinsing several times EtOAc. The filtrate was washed with saturated NaHCO3saturated salt solution, dried over MgSO4and concentrated in vacuum. Crude product was passed through a small layer of silica gel, elwira a mixture of hexane/EtOAc 4:1, receiving of 17.1 mg (yield 99%) 71a in the form of a colorless oil:1H NMR (400 MHz, CDCl3) δ (m, 6H), is 0.96 (t, J=7.9 Hz, 9H), of 0.97 (d, J=6.8 Hz, 3H), of 1.05 (d, J=6.8 Hz, 3H), 1,11 (c, 3H), 1.26 in (c, 3H), 1,44 (c, 9H), 1,84-1,90 (m, 1H), 2,21 (DD, J=6,7 and 17.0 Hz, 1H), a 2.36 (DD, J=6,7 and 17.0 Hz, 1H), 3,24-3,29 (m, 1H), 3,44-to 3.52 (m, 2H), to 3.67 (DD, J=3,9 and 8.9 Hz, 1H), 4,36 (DD, J=3.5 and 6.5 Hz, 1H), 4,50 (d, J=12.0 Hz, 1H), 4,54 (d, J=12.0 Hz, 1H), 7,32 and 7.36 (m, 5H);13With NMR (100 MHz, DCl3) δ 5,0, 6,9, 9,7,13,9, 20,2, 21,8, 28,0, 36,3, 40,8, 41,5, 53,7, 72,5, 72,9, 73,2, 73,6, 80,7, 127,4, 127,5, 128,2, 138,6, 171,0, 221,4; IR (film, NaCl, cm-1) 3502, 2959, 2875, 1731, 1683, 1456, 1366, 1154, 1098, 996, 739; MS-HP (ESI) calculated for C30H52About6SiCl3Na [M+Na+] 559,3 found 559,3; [α]23D= -41,0 (c = 0.4, CHCl3).

Tert-butyl ester 9-benzyloxy-7-(tert-butyldimethylsilyloxy)-3-(diethylethylenediamine)-4,4,6,8-tetramethyl-5-oksanalove acid (36)

To a solution of 71a (4,1 is g, 7.6 μmol) and 2,6-lutidine (10,0 ál, to 43.5 mmol) in CH2Cl2(0.2 ml) at -78°C was added TBSOTf (10,0 ál, to 85.8 mmol). After 2 h addition was added 2,6-lutidine (10,0 ál, to 43.5 mmol) and TBSOTf (10,0 ál, to 85.8 mmol). After 6 h the mixture was diluted with saturated aqueous NaHCO3. The aqueous layer three times was extracted with EtOAc. The combined organic extracts were washed with saturated salt solution, dried over MgSO4and concentrated under reduced pressure. Crude product was purified flash chromatography (gradient from hexane to a mixture of hexane/EtOAc 91:9)to give 36 (5.4 mg, 82%) as a clear oil. Spectroscopic data are in good agreement with the described values.

Alcohol 83.To a solution of ethyl 4,4,4-triftoratsetata (24,0 ml, 0,164 mol) in a mixture of THF-water (3:1 = V:V, 320 ml) at room temperature was added allylbromide (20,0 ml, 1.4 EQ.) and indium (powder, -100 mesh, 25 g, 1.3 EQ.) and the resulting mixture was stirred at 48°C for 15 hours. The reaction mixture was cooled to room temperature, extinguished 2N aqueous HCl (400 ml) and was extracted with CH2Cl2(400 ml, 2×200 ml). The combined organic layers were dried (MgSO4), filtered and concentrated in vacuum. Flash chromatography (hexane → hexane-ether 10:1 → 8:1 → 6:1 → 4:1) gave alcohol 83 in the form of a clear oil (31,64 g, yield 85%): IR (film) 3426 (osirm), 2986 (m), 1713 (c), 1377 (m),1345 (m), 1301 (m), 1232 (m), 1173 (c), 1095 (m), 1023 (m), 927 (m) cm-1;1H NMR (400 MHz, CDCl3) δ of 5.82 (m, 1H), 5,15 (m, 3H), 4,17 (m, 2H), 2,59 (m, 1H), 2,58 (d, J=3,4 Hz, 2H), to 2.29 (DD, J=14,2, 8.6 Hz, 1H), 1,24 (t, J=7.2 Hz, 3H);13With NMR (100 MHz, DCl3) δ 172,08, 130,89, 125,65 (kV, J=280 Hz), 120,27, 73,79 (kV, J=28 Hz), 61,55, 38,97, 35,65, 13,82; the mass spectrum of the high-resolution m/z 227,0895 [(M+H)+; calculated for C9H14About3F3: 227,0895].

Ester 84. The mixture of alcohol 83 (16,71 g, 0,07386 mol) and pyridine (15.0 ml, 2.5 EQ.) was cooled to -10°C and slowly over 11 min were treated with thionyl chloride (11.3 ml, 2.1 EQ.). The resulting mixture was heated to 55°C and was stirred for 12 hours. The reaction mixture was cooled to -5°C, extinguished with water (200 ml) and was extracted with CH2Cl2(2×200 ml, 2×150 ml). The combined organic layers were washed with saturated NaHCO3(2×200 ml) and saturated salt solution (200 ml), dried (MgSO4) and concentrated in vacuum. Flash chromatography (pentane:ether 15:1) gave ester 84 (11,90 g, yield 77%) as a yellow oil: IR (film) 2986 (w), 1731 (c), 1308 (c), 1265 (w), 1227 (m), 1197 (c), 1133 (c), 1025 (m), 920 (w), 896 (w) cm-1;1H NMR (400 MHz, CDCl3) δ 6,36 (c, 1H), 5,79 (DDT, J=16.9 and, of 10.2, 6.6 Hz, 1H), 5,15 (DD, J=17,1, 1.5 Hz, 1H), 5,08 (DD, J=10,0, 1.4 Hz, 1H), 4,22 (kV, J=7,1 Hz, 2H), 3,44 (d, J=6,5 Hz, 2H), 1,29 (t, J=7,1 Hz, 3H);13With NMR (100 MHz, DCl3) δ 164,22, 143,37 (kV, J=29 Hz), 132,71, 123,21 (kV, J=274 Hz), 122,60 (kV, J=6 Hz), 117,32, 60,85, 30,54, 13,85; m is SS-spectrum high-resolution m/z 209,0788 [(M+H) +; calculated for C9H12About2F3: 209,0789].

Alcohol 85.To a cooled (-75°C) solution of ester 84 (7,12 g, 0,0342 mol) in CH2Cl2(120 ml) was added a solution of DIBAL-H (75 ml, 2.2 EQ.) in CH2Cl2(1.0 M) and the resulting mixture was heated to room temperature within 3 hours. The reaction mixture was cooled to 0°C, reduce saturated NH4Cl (12 ml) and stirred at room temperature for 20 minutes, the Reaction mixture was diluted with ether (200 ml), dried (MgSO4) and concentrated in vacuum. Flash chromatography (pentane:ether, 3:1-1:1) gave the alcohol 85 (of 5.68 g, 99%) as a clear oil: IR (film) 3331 (users), 2929 (m), 1642 (m), 1445 (m), 1417 (w), 1348 (c), 1316 (c), 1217 (c), 1175 (c), 1119 (c), 1045 (m), 985 (c), 921 (m), 831 (w) cm-1;1H NMR (400 MHz, CDCl3) δ 6,33 (TD, J=6,1, 1,6 Hz, 1H), of 5.75 (DDT, J=17.2 in, 10,0, 6.2 Hz, 1H), 5,07 (m, 2 H), the 4.29 (DDD, J=6,3, 4,3, 2,1 Hz, 2H), 2,95 (d, J=6.2 Hz, 2H);13With NMR (100 MHz, DCl3) δ 134,45 (kV, J=6 Hz), 133,38, 127,97 (kV, J=29 Hz), 123,76 (kV, J=271 Hz), 116,25, 57,87, 29,79.

Iodide 86.A cooled (0°C) solution of alcohol 85 (5,97 g, 0,0358 mol) in CH2Cl2(50 ml) was treated with PPh3(11,17 g, 1.2 EQ.), the imidazole (3.55 g, 1.5 EQ.) and I2(9,10 g, 1.1 EQ.) and the resulting mixture was stirred at 0°C for 10 min, the Reaction mixture was extinguished with a mixture of saturated Na2S2O3- n is Semenovo NaHCO 3(1:1 = V:V, 200 ml) and was extracted with pentane (3×200 ml). The combined organic layers were washed with a mixture of saturated Na2S2O3- saturated NaHCO3(1:1 = V:V, 200 ml) and saturated salt solution (100 ml), dried (MgSO4) and concentrated in vacuum. Flash chromatography (pentane) gave the iodide 86 (6,69 g, 68%) as a pale red oil: IR (film) 3083 (w), 2982 (w), 1636 (w), 1558 (w), 1456 (w), 1367 (w), 1317 (c), 1216 (m), 1181 (c), 1151 (c), 1120 (c), 989 (m), 921 (m), 896 (m) cm-1;1H NMR (400 MHz, CDCl3) δ 6,45 (TD, J=8,9, 1.5 Hz, 1H), 5,79 (DDT, J=16,8, 10,3, 6.2 Hz, 1H), 5,12 (m, 2H), 3,85 (DDD, J=8,9, of 2.9, and 1.4 Hz, 2H), 3.00 and (dt, J=6,1, 1,4 Hz, 2H);13With NMR (100 MHz, DCl3) δ 132,42, 131,64 (kV, J=6 Hz), 129,63 (kV, J=29 Hz), 123,64 (q, J=272 Hz), 117,00, 29,32, -4,27; mass spectrum high-resolution m/z 298,7 [(M+Na)+; calculated for C7H8F3INa: 299,0].

α-Hydroxycytidine 88.To a cooled (-78°C) TES-protected 4-benzyl-3-hydroxyacetanilide-2-ONU 7 (16.28 per g of 1.92 EQ.) in THF (160 ml) dropwise over 51 min solution was added LHMDS (42,0 ml, 1,73 EQ.) in THF (1.0 M) and the resulting mixture was stirred at -78°C for 35 minutes, the Reaction mixture was treated with a solution of iodide 86 (6,69 g, and 24.2 mmol) in THF (10 ml) and the resulting mixture gave the opportunity to slowly warm to room temperature over night. The reaction mixture was extinguished saturated NAHCO3(200 ml), and ek is was tragically EtOAc (3×200 ml). The combined organic layers were washed with saturated NH4Cl (150 ml), saturated salt solution (150 ml), dried (MgSO4) and concentrated in vacuum. Flash chromatography (hexane-EtOAc 6:1 → 3:1) gave a mixture of products of alkylation (13,6 g), which was used in the next reaction without further purification. The solution products of alkylation in a mixture of HOAc-water-THF (3:1:1 = V:V:V, 200 ml) was stirred at room temperature for 4 hours. The reaction mixture was concentrated in vacuum to remove HOAc, reduce saturated NaHCO3(400 ml) and was extracted with EtOAc (3×200 ml). The combined organic layers were dried (MgSO4) and concentrated in vacuum. Flash chromatography (hexane : EtOAc, 3:1 → 2:1) gave the α-hydroxycytidine 88 (of 7.55 g, yield 81% for two steps) as a clear oil: [α]D25-48,2 (1,08, l3); IR (film) 3486 (users), 3030 (m), 2983 (c), 2925 (m), 1790 (c), 1682 (c), 1481 (m), 1393 (m), 1360 (m), 1217 (m), 1171 (m), 1113 (m), 992 (m), 919 (m), 847 (w) cm-1;1H NMR (400 MHz, CDCl3) δ 7,32 (m, 3H), 7,17 (m, 2H), 6,33 (TD, J=7,2, 1.5 Hz, 1H), 5,77 (DDT, J=16,6, 10,1, 6.2 Hz, 1H), 5,08 (m, 3H), 4,74 (DDT, J=4,8, and 3.7, and 4.4 Hz, 1H), 4,33 (DD, J=8,6, 8.6 Hz, 1H), 4.26 deaths (DD, J=9,2, 3,4 Hz, 1H), 3,42 (userd, J=6,4 Hz, 1H), 3,24 (DD, J=13,5, and 3.4 Hz, 1H), 2,99 (m, 2H), and 2.79 (DD, J=13,5, and 9.4 Hz, 1H), 2,70 (m, 1H), 2,50 (m, 1H);13With NMR (125 MHz, DCl3) δ 173,93, 153,05, 134,43, 133,64, 129,98 (kV, J=6 Hz), 129,82 (kV, J=28 Hz), 129,29, 120,01, 127,58, 124,00 (kV, J=272 Hz), 116,34, 69,60, 67,31, 54,95, 37,78, 32,29, 29,84; mass spectrum you what the resolution will be about m/z 384,1421 [(M+N) +; calculated for C19H21NO4F3: 384,1423].

α-Hydroxyamide 89.A suspension of (MeO)NHMe·HCl (10.1 g, 5,25 EQ.) in THF (100 ml) at 0°C was treated dropwise with a solution AlMe3(50 ml, 5.1 EQ.) in toluene (2.0 M) and the resulting clear solution was stirred at room temperature for 34 min, then was added to a cooled (0) solution of α-hydroxyacetophenone 88 (of 7.55 g of 19.7 mmol) in THF (70 ml). The resulting mixture was heated to room temperature and was stirred for 12 hours. The reaction mixture was cooled to 0°C, was suppressed by slow addition of 1N aqueous tartaric acid (100 ml)was stirred at room temperature for 25 min and was extracted with EtOAc (3×200 ml). The combined organic layers were dried (MgSO4) and concentrated in vacuum. Flash chromatography (hexane:EtOAc, 2:1 → 1:1) gave the α-hydroxyamide 89 (5,12 g, yield 97%) as a clear oil: [α]D25-57,2 (1,03, l3); IR (film) 3432 (users), 3084 (w), 2980 (m), 2943 (m), 1652 (c), 1464 (m), 1373 (m), 1318 (m), 1214 (m), 1171 (m), 1112 (m), 991 (m), 919 (m), 818 (w) cm-1;1H NMR (400 MHz, CDCl3) δ 6,32 (TD, J=7,3, 1.5 Hz, 1H), grade of 5.74 (DDT, J=16.9 and, 10,3, 6,1 Hz, 1H), of 5.05 (m, 2H), 4,43 (DD, J=7,6, 3.5 Hz, 1H), 3,70 (c, 3H), 3,35 (users, 1H), 3,24 (c, 3H), 2,94 (d, J=6,1 Hz, 2H), 2,59 (m, 1H), a 2.36 (m, 1H);13With NMR (100 MHz, DCl3) δ 173,43, 133,68, 130,59 (kV, J=6 Hz), 129,25 (kV, J=28 Hz), 124,05 (kV, J=271 Hz), 116,17, 67,57, 61,44, 2,56, 32,38, 29,75; mass spectrum high-resolution m/z 268,1161 [(M+N)+; calculated for C11H17NO3F3: 268,1161].

α-Hydroxyketone 90.To a cooled (0°C) solution of α-hydroxyamide 89 (4,87 g, 18.2 mmol) in THF (150 ml) was added a solution of MeMgBr (75 ml, 12 EQ.) in simple ether (3.0 M). After 5 min the reaction mixture was extinguished saturated NH4Cl (250 ml) and was extracted with EtOAc (5×200 ml). The combined organic layers were dried (MgSO4) and concentrated in vacuum. Flash chromatography (hexane:EtOAc 4:1 → 2:1 → 1:2) gave α-hydroxyketone 90 (2.16 g, yield 53%, the yield was 73% based on retrieval of the original substance) in the form of a clear oil and the starting material α-hydroxyamide 89 (1,30 g, yield 27%): [α]D25+58,5 (1,30, l3); IR (film) 3460 (users), 3085 (w), 2984 (m), 2926 (m), 1716 (c), 1679 (m), 1641 (m), 1417 (m), 1361 (m), 1319 (c), 1247 (m), 1216 (c), 1172 (c), 1113 (c), 1020 (m), 994 (m), 968 (w), 919 (m) cm-1;1H NMR (500 MHz, CDCl3) δ 6,21 (t, J=7,0 Hz, 1H), of 5.75 (DDT, J=16,7, 10,4, 6.2 Hz, 1H), 5,07 (m, 2H), 4.26 deaths (dt, J=7,1, 4.5 Hz, 1H), 3,51 (d, J=4,7 Hz, 1H), 2,96 (d, J=6,1 Hz, 2H), 2,66 (m, 1H), 2,42 (m, 1H), 2,19 (c, 3H);13With NMR (100 MHz, DCl3) δ 208,53, 133,43, 129,80 (kV, J=28 Hz), 129,76 (kV, J=6 Hz), 123,85 (kV, J=271 Hz), 116,32, 75,36, 31,22, 29,81, 25,11; the mass spectrum of the high-resolution m/z 223,0945 [(M+N)+; calculated for C10H14NO2F3: 223,0946].

Example 8

Method for catalytic asymmetric oxidation

Example 9

Synthesis of 21-amino-26 trifter-(E)-9,10-degidro-dEpoB

Compound 98:To a solution of 59 (of 50.4 mg, 90,1 mmol) in THF (1 ml) was added (PhO)2PON3to 27.2 μl, 126 mmol) at 0°C. After stirring at 0°C for 5 min was added DBU (16,2 μl, 108 mmol). After stirring at 0°C for 2 h the mixture was stirred at RT for 20,5 hour. The reaction mixture was diluted with EtOAc and was suppressed by the addition of water (2 ml). After separation of the layers the aqueous layer was extracted with EtOAc (three times) and the combined organic layers were dried over Na2SO4. After concentration the residue was dried under high vacuum for 10 min to remove DBU. Purification with flash chromatography on a column (SiO2, hexane/EtOAc = 3:2) gave the azide 98 (45,6 mg, 78,0 mmol, 87%) as colorless solids;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,8 Hz)of 1.33 (3H, c), a 2.01 (1H, d, J=5.5 Hz, OH), 2,17 (3H, c), of 2.25 to 2.35 (1H, m), is 2.41 (1H, DD, J=15,5, and 3.2 Hz), 2.49 USD (1H, DD, J=15,5, 9.5 Hz), 2,54-2,60 (1H, m), 2,66 (1H, d, J=6.0 Hz), 2,65 was 2.76 (1H, m), 2,96 (1H, DD, J=16,0, 4,2 Hz), 3,03 (1H, DD, J=16,1, 6,7 Hz), 3,11 (1H, quintet, J=6.8 Hz), 3,71 is 3.76 (1H, m), or 4.31 (1H, DDD, J=9,2, of 5.9, 3.2 Hz)and 4.65 (2H, c), 5,43 (1H, DD, J=6,0, a 4.3 Hz), to 5.58 (1H, DDD, J=15,8, 6,4, 4.6 Hz), to 5.66 (1H, DD, J=15,8, 6,1 Hz), 6,23 (1H, t, J=7,3 Hz), 6,63 (1H, c), 7,18 (1H, c); MS-HP (ESI) calculated for C27H35F3N4O5SNa[M+Na+] 607,2 found 607,2

Compound 96:To a solution of azide 98 (21,0 mg, or 35.9 mmol) in THF (0.6 ml) was added PMe3(1.0 M in THF, 43,1 ál, to 43.1 mmol). After stirring at RT for 2 min was added water (0.1 ml) and the mixture was stirred at RT for 3 hours. Added PMe3(1.0 M in THF, to 7.2 μl, 7.2 mmol) and the mixture was stirred at RT for 1.5 hour. To the mixture was added 28% NH4OH (water) (54,5 µl). After stirring for 1 hour the mixture was directly purified preparative TLC (CH2Cl2/MeOH = 100:7,5), receiving Amin 96 (to 15.9 mg, 28.5 μmol, 79%) as colorless solids;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,8 Hz)of 1.34 (3H, c), 2,12 (3H, d, J=0.7 Hz), 2,24 to 2.35 (1H, m), 2,39 (1H, DD, J=15,4, 3.0 Hz), 2.49 USD (1H, DD, J=15,4, 9.8 Hz), 2,54-2,63 (1H, m), 2,66-was 2.76 (1H, m), of 2.97 (1H, DD, J=16,2, 4,2 Hz), 3,03 (1H, DD, J=16,3, 6.5 Hz), 3,10 (1H, quintet, J=6.8 Hz), 3,74 (1H, DD, J=6,7, 3.5 Hz), 4,18 (2H, c), 4,34 (1H, DD, J=9,8, 2,9 Hz), 5,43 (1H, DD, J=6,0, a 4.3 Hz), 5,55-5,64 (1H, m), 5,67 (1H, DD, J=15,9, 5.8 Hz), 6,24 (1H, ushort, J=7,3 Hz), 6,66 (1H, c), 7,10 (1H, c); MS-HP (ESI) calculated for C27H38F3N2O5S [M+N+] 559,2 found 559,2.

Compound 97:To a solution of amine 96 (15,9 m, 28.5 mmol) in CH3CN (0,78 ml) was added 37% HCHO (water) (31,4 μl, 0,143 mmol), then NaBH3CN (1.0 M in THF, to 85.5 μl, of 85.5 mmol) and the mixture was stirred at RT for 20 min was Added AcOH (1 drop) and the mixture was stirred at KTV for 40 minutes The mixture was purified directly by preparative TLC (CH2Cl2/MeOH = 100:8)to give the product 97 (15.6 mg, to 26.6 mmol, 93%) as colorless solids;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=6,9 Hz)of 1.23 (3H, d, J=6,8 Hz)of 1.33 (3H, c), 2,17 (3H, c), 2,24 to 2.35 (1H, m), 2,43 (1H, DD, J=15,7, 3.6 Hz), 2.49 USD (1H, DD, J=15,6, and 9.1 Hz), 2,55-of 2.64 (2H, m, including HE), 2,68-2,77 (1H, m), 2,80 (3H, c), of 2.81 (3H, c), 2,92-of 3.06 (2H, m), 3,10 (1H, quintet, J=6.8 Hz), 3,69 is 3.76 (1H, m), 4,25-4,34 (1H, m)to 4.33 (2H, c), 5,42 (1H, t, J=5.5 Hz), to 5.57 (1H, dt, J=15,8, 6.3 Hz), to 5.66 (1H, DD, J=15,7, 6.4 Hz), to 6.22 (1H, ushort, J=7,2 Hz), only 6.64 (1H, c), 7,30 (1H, c); MS-HP (ESI) calculated for C29H42F3N2O5S [M+H+] 580,2 found 580,2.

Connections 94 and 95:To a mixture of 59 (18,9 mg, 33.8 mmol) and Et3N (18,8 μl, is 0.135 mmol) in CH2Cl2(1 ml) was added TsCl (12.9 mg, of 67.5 mmol) and DMAP (2.1 mg, about 16.9 mmol) at 0°C. After stirring at RT for 1.5 h the mixture was diluted with EtOAc and filtered through a bed of silica gel (EtOAc washing). After concentration the residue was purified preparative TLC (hexane/EtOAc = 1:1), getting toilet 94 (8.5 mg, to 11.9 mmol, 35%) and chloride 95 (4.3 mg, 7,44 mmol, 22%), both as colorless solids;

1H NMR (400 MHz, CDCl3) δ of 1.06 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,7 Hz)of 1.33 (3H, c), 1,99 (1H, d, J=5.5 Hz), 2,10 (3H, c), 2,25-of 2.34 (1H, m) to 2.41 (1H, DD, J=15,5, and 3.3 Hz), 2,47 (3H, c), 2,48 (1H, DD, J=15,7, and 9.4 Hz), of 2.51 2.63 in (1H, m), 2.63 in (1H, d, J=6,1 Hz, OH), 2,64 is 2.75 (1H, m),2.91 in was 3.05 (2H, m)3,10 (1H, quintet, J=6.8 Hz), 3,70 of 3.75 (1H, m), 4,30 (1H, DDD, J=9,3, a 6.1, a 3.2 Hz), 5,32 (2H, c), 5,41 (1H, DD, J=5,8,4,5 Hz), to 5.57 (1H, DDD, J=15,8, 6,4, 4.6 Hz), the 5.65 (1H, DD, J=15,8, 6,0 Hz), 6,21 (1H, t, J=7,1 Hz), 6,59 (1H, c), 7,18 (1H, c), 7,37 (2H, d, J=8.1 Hz), to 7.84 (2H, d, J=8,3 Hz); MS-HP (ESI) calculated for C34H42F3NO8S2Na[M+Na+] 736,2 found 736,3.

1H NMR (400 MHz, CDCl3) δ of 1.06 (3H, c), of 1.12 (3H, d, J=6,9 Hz)of 1.23 (3H, d, J=6,7 Hz)of 1.34 (3H, c), 2,00 (1H, d, J=5.6 Hz, HE), OF 2.15 (3H, c), of 2.25 to 2.35 (1H, m), is 2.41 (1H, DD, J=15,5, and 3.2 Hz), 2.49 USD (1H, DD, J=15,5, and 9.4 Hz), 2,53-2,62 (1H, m), 2,69 (1H, d, J=6,1 Hz, HE), 2,66 was 2.76 (1H, m), 2,92 was 3.05 (2H, m), 3,11 (1H, quintet, J=6.4 Hz), 3,70 is 3.76 (1H, m), 4,32 (1H, DDD, J=9,2, 5,9, 3.1 Hz), 4,85 (2N, c), 5,43 (1H, DD, J=6,0, 4,4 Hz), 5,59 (1H, DDD, J=15,9, 6,4, and 4.5 Hz), to 5.66 (1H, DD, J=15,9, 6,1 Hz), 6,23 (1H, t, J=6.8 Hz), 6,63 (1H, c), 7,20 (1H, c); MS-HP (ESI) calculated for C27H35ClF3NO5Sna [M+Na+] 600,2 found 600,2.

Compound 99:To a solution of 59 (6.9 mg, 12.3 mmol) in CH2Cl2(0.4 ml) was added activated MnO2(purchased from Acros, and 26.8 mg, 0,308 mmol). After vigorous stirring at RT for 4 h the mixture was filtered through a pad celite, which was washed EtOAc. After concentration the residue was purified preparative TLC (hexane/EtOAc = 1:1)to give aldehyde 99 (2.7 mg, 4,84 μmol, 39%) as colorless solids;1H NMR (400 MHz, CDCl3) δ of 1.06 (3H, c), of 1.13 (3H, d, J=7,2 Hz), 1,24 (3H, d, J=6.9 Hz), of 1.35 (3H, c), a 1.96 (1H, d, J=5.6 Hz, HE), 2,2 (3H, d, J=0.7 Hz), of 2.25 to 2.35 (1H, m)2,44 (1H, DD, J=15,4, 3.5 Hz), 2,46 (1H, d, J=5,9 Hz, OH), of 2.51 (1H, DD, J=15,7, and 9.3 Hz), 2.57 m-28 (1H, m), 2,68-and 2.79 (1H, m), 2,96-3,03 (2H, m), 3,10 (1H, quintet, J=6.8 Hz), 3,71-3,76 (1H, m), or 4.31 (1H, DDD, J=9,4, 6,3, 3.5 Hz), the 5.45 (1H, t, J=5.0 Hz), of 5.53-5,63 (1H, m), 5,67 (1H, DD, J=15,7, 6.2 Hz), 6,24 (1H, t, J=6.6 Hz), 6,72 (1H, c), EUR 7.57 (1H, d, J=0.9 Hz), 10,01 (1H, d, J=1.2 Hz).

Compound 100:To a solution of aldehyde 99 (4.6 mg, 8.25 mmol) in CH3CN (0.5 ml) at 0°C was added MeNH2(2.0 M in THF, 41,3 µl, a 41.3 mmol). After stirring at 0°C for 15 min was added NaBH3CN (1.0 M in THF; 25 μl; 25 Microm). After stirring at 0°C for 0.5 h was added AcOH (3 drops). After stirring at 0°C for 2 hours was added 28% NH4OH (water)(40 μl) and the mixture was stirred at RT for 10 minutes the Mixture directly twice was purified preparative TLC (CH2Cl2/MeOH = 100:9), receiving 100 (2.4 mg, 4,19 mmol, 51%) as colorless solids;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,8 Hz)of 1.34 (3H, c)to 2.13 (3H, c), 2,25-of 2.34 (1H, m), 2,39 (1H, DD, J=15,3, 3.0 Hz), 2.49 USD (1H, DD, J=15,3, 9.7 Hz), of 2.56 (3H, c), 2,54-of 2.64 (1H, m), 2,66 is 2.75 (1H, m), 2,89 (1H, d, J=5,1 Hz), 2,94 was 3.05 (2H, m), 3,11 (1H, quintet, J=6.8 Hz), 3,74 (1H, DD, J=6,6, 3.5 Hz), 4,08 (2H, c), 4,34 (1H, DD, J=9,6, 2,9 Hz), 5,43 (1H, DD, J=6,2, 4,1 Hz), 5,56-5,63 (1H, m), to 5.66 (1H, DD, J=15,9, 5.7 Hz), 6,24 (1H, t, J=7,3 Hz), 6,66 (1H, c), 7,11 (1H, c); MS-HP (ESI) calculated for C28H40F3N2O5S [M+H+] 573,3 found 573,3.

Example 10

Analogization, which lead to ablation xenotransplantion tumors to Prezidiuma status

By combining chemical synthesis, molecular modeling and spectroscopic analyses, the authors found that the introduction of E-9,10-double bond (see connection 28 below) gives approximately a 10-fold increase in the efficiency of the drug in the experiments on the basis of xenografts using tumor MX-1, multidrug-resistant (A. Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899; included in this description by reference). After mapping experiments in vitro and in vivo, aimed at the tumor type, MX-1, it became apparent that the 28 is more cytotoxic than 2b. However, another factor that contributes is that the rest of the lactone in a series of 9,10-dihydrocodeinone is significantly more stable in plasma of mice and man, than in the case of 9,10-dehydropeptidase the same series. The result of these two additional effects were giving 28 ability to perform complete suppression of tumors in multiple xenografts at a dose of 3 mg/kg, in contrast to schemes with a dose of 30 mg/kg in the case of 1.

When the termination processing palpable tumors appear again some animals. Thus, at least at the present time fully synthetic is the cue 28 does not fully comply with the stringent standards highly preferred effective therapeutic index and elimination of tumors to Prezidiuma state.

The data drew the authors ' attention to the consequences of the replacement of three hydrogen atoms 26-methyl group 28 three fluorine atoms. The introduction of fluorine atoms in this situation leads to increased stability of the 12,13-double bond with respect to oxidation (Smart, B. E. J. Fluorine Chem. 2001, 109, 3; publication are included in this invention in the form of links). Previous experience testified to some weakening of cytotoxicity when replacing the polar groups in the field of C12-C13 double bond (A. Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899; publication are included in this description by reference). In this description, the authors report the receipt by total chemical synthesis of 9,10-degidro-26-triftoretil, especially focusing on the unique biological activity of the original structure 29.

Carefully studied therapeutic efficacy of dEpoB 30 mg/kg), paclitaxel (20 mg/kg) and F3-deH-dEpoB (29; 20 and 30 mg/kg) against xenografts of breast carcinoma human MX-1 in relation to the disappearance of the tumors and relapse, and the results are shown in table 10-1. Each dosing group consisted of four or more of nude mice. Weight refers to the total body weight minus the weight of the tumor. In the case of all three compounds was achieved by the disappearance of the tumors. On the 10th day after the end of treatment relapse occurred in 5/10 (dEpoB),
2/7 (Pak who taxel) and 0/4 (compound 29) mice. Extended follow-up after discontinuation of treatment with doses 29, constituting 20 mg/kg, showed a long-term absence of tumors up to 27 days, and at this time, 2 of the 4 tumors of mice had anticipated. It is noteworthy that treatment with doses 29, constituting 30 mg/kg resulted in complete disappearance of the tumors and the absence of any relapse within two months after stopping treatment.

Table 10-1
Therapeutic effect of dEpoB, paclitaxel and F3-deH-dEpoB against xenograft MX-1 in mice nude[a]
Drug
tool
Dose
(mg/kg)
Changes in body mass (%)The tumor was removed after Q2Dx6 6 hour/in infusionThe tumor appeared again
on day 10 after injection
On day 4
after termination of the introduction of
On the 8th day after the termination of the introduction of
dEpoB (1)30-25,3±2,1-9,1±4,110/10 5/10
Paclitaxel20-23,9±3,7-8,7±0,77/72/7
F3the deH- 20-22,4±0,6is 7.3±0,74/40/4[b]
dEpoB (29)30-27,1±2,7-17,4±5,54/40/4[b]
[a]50 mg tissue xenograft breast carcinoma human MX-1 implanted p/in 0 day. Treatment Q2Dx6 using 6-hour/in infusion was started on day 8 and was completed on the 18th day.
[b]Registered reappearance of a tumor in 2/4 on the 27th day after the end of treatment. Was not observed further re-occurrence of tumors within 28-64-th days after discontinuation of treatment.
[c]No re-occurrence of tumors within 64 days after stopping treatment when he was finished the experiment.

Dose reduction agent 29 to 10 mg/kg (Q2D) also led to the disappearance of the tumor MX-1, but to achieve this result is the required nine doses (Fig, 58 and 59A). As an additional check chemotherapeutic treatment was delayed until the tumor size reached 0.5 g (~2.3% of body weight). Treatment doses 29, constituting 25 mg/kg (Q2Dx7), caused the disappearance of 4/4 tumors in mice. In contrast, the required dose of dEpoB, comprising 30 mg/kg (Q2Dx8), in order to induce the disappearance of tumors in 3 of 4 mice. However, unlike the case with the use of 29 the apparent disappearance, which occurred after treatment dEpoB, suffered relapses over time (pigv).

The fact that the agent 29 completely inhibited the growth of xenografts of breast carcinoma human MX-1, reduced the tumor and cause them to disappear for such a long period of time as 64 days, makes a deep impression. Moreover, after the course of treatment carried out with 29 (20 mg/kg or 30 mg/kg Q2Dx6, 6-hour/in infusion, table 1 above) body weight of organisms with xenografts returned to control levels before treatment within 12-18 days after stopping treatment. The specified observation indicates the absence of damage to vital organs. At low therapeutic doses of 10 mg/kg, Q2DX12 (pigv) the maximum weight loss was only 12% weight gain by 6% during the last three doses. Body weight returned to control urovnya treatment just three days after stopping treatment. Table 1 above shows that the animals can survive the loss of body weight up to 27%. Achieved in this case the stock of therapeutic security is surprisingly wide for therapeutic antitumor therapeutic agent.

Also assessed therapeutic efficacy 29 against xenograft of human lung carcinoma (A549) and xenografts resistant to paclitaxel of human lung carcinoma A549/Taxol (figs and 59D). Slowly growing xenografts lung carcinoma A549 was treated 29 (25 mg/kg, Q2DX6, twice, eight days separately), and this led to 99.5% suppression of tumors in the end with the complete eradication 4 of 4 tumors after two additional doses (figs). Interestingly, body weight of mice was decreased to 35% without any mortality and cessation of treatment resulted in rapid recovery of body weight almost to the control level before treatment (figs). In contrast, a parallel study using dEpoB 30 mg/kg, Q2Dx6) resulted in suppression of 97.6% of tumors, but did not lead to the eradication of tumors. Additional research 29 (dose 20 mg/kg) compared to resistant to the xenograft A549/Taxol (fig.59D) tumor growth was completely suppressed, and the tumor eventually decreased by 24.4% from control to treatment. In this study the maximum body weight was reduced by 24%, however is after stopping treatment drug body weight returned to 90% of control before treatment. In the comparative study (E)-9,10-degidro-dEpoB (28, a group of 4 mg/kg) tumor growth was reduced by 41.6%.

Data relevant to the analysis of what factors provide a striking therapeutic index of the compound 29, along with comparable data related to cognate representatives of this series, presented in table 10-2. It is noted that in relation to the inherent cytotoxicity in the transition from EpoB (2b) to dEpoB (1) lost a whole order of magnitude. About 60% of the said loss is reversed if 9,10-degidro-dEpoB (28). Of a specified inherent cytotoxicity is lost in the transition to the connection 29, which is in the cell toxicity has at least ~1.8 times higher than in the case of a reference compound dEpoB.

The authors note that among 12,13-dehydroemetine, 29 shows much better stability in plasma of mice, and is also the most stable in plasma human liver S9. The authors also note that in 2-sets 12,13-digidrosteron 26 trifter-containing sample brings a lower lipophilicity, and to some extent increased solubility in water (table 10-2, below). Now it may be clear that a great advantage 29 is the result of increased stability in serum and bioavailability.

Table 10-2
The profile derived dEpoB
ConnectionCytotoxic-ical efficiency of IC50(nm)[a]Maxim-aspects
reduction of body weight in % without loss
The stability, half-lifeDissolve confidence in water (µg/ml)The lipophilicity, the distribution of the octanol/water (POW)Scheme therapeutic dosing for Q2D 6 hours/infusion (mg/kg)Relative therapeu optical index MTD[b]
Plasma of mice (min)The S9 fraction of liver human (h)
EpoB (2b)0,53±0,2155715,8NDND0,6-0,8+++
dEpoB (1)5,6±2,83246±71,0±0,19,4/td> 4,425-30++++
deH-dEpoB (28)0,90±0,402984±64,9±0,7273,33-4++++
F3-dEpoB (2)9,3±5,22266±71,6±0,484,115-20++
F3-deH-dEpoB (29)3,2±0,333212±8810,5±2,3203,310-30+++++
[a]The values of the IC50see for leukemia cells CCRF-CEM. Values are based on the number of values from the two experiments; all values are derived from the seven points of concentration; ND = not determined.
[b]Ranked relative therapeutic index (TI) at the MTD (maximum permissible dose):
+ Tumor growth is davlen by 25-50%.
++ Tumor growth suppressed by 50-100%.
+++ The tumor is reduced, but not eliminated.
++++ the tumor disappears in some or all of nude mice with slow recovery of body weight and/or relapse in some mice within one week after stopping treatment.
+++++ The tumor disappeared in all nude mice, body weight quickly recovered and/or another relapse.
Therapeutic trial epothilones against xenografts of nude mice, such as the MX-1, were studied Chou, T. C. et al. Proc. Natl. Acad. Sci. USA. 1998, 95, 15798, and in 2001, 98, 8113.

All agents 1-2 and 28-29 for the first time obtained using the General synthesis. Practical synthesis of 1 previously described (Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899; White et al. J. Am. Chem. Soc. 2001, 123, 5407; Yoshimura et al. Angew. Chem. 2003, 42, 2518; Rivkin et al., J. Org. Chem. 2002, 124, 7737; each publication included in this description by reference). Also first described ways to create 28 and 29 relating to inventive step. Selective restoration of the 9,10-double bond 29 gave 2. The striking results obtained on the basis of the above studies xenografts in relation to the most promising at the present time connection 29 will undoubtedly require further advance its detailed Toxicological and pharmacokinetic studies in higher animals and from them, if appropriate, the promotion is ment to clinical trials in humans. Such prospects have completely changed the nature of the problem of synthesis from receipt of samples to the problem of obtaining quantities of these new epothilone agents, as measured by many grams. Made an important modernization of the previous methods authors, first conceived and shown in terms of carrying out the invention. In particular, the new protocols, the authors carried out a large simplifications in the stereospecific construction in relation to the carbon atoms 3 and 26. Alcohol 32 receives, as described previously (Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899; included in this description by reference). It will be noted that in the case of a new synthesis stereocenter 6, 7 and 8 are obtained from obviously available ketone 30 and aldehyde 31. After protection of the alcohol and hydrolysis of the acetal corresponding aldehyde are condensed with tert-butyl acetate, receiving product, such alcolu. Since this condensation is not controlled in the ratio of diastereomers was required and implemented corrective measure. Oxidation of this mixture 1:1 C3-epimeres gave ketone 69. After a very successful repair Noyori (Noyori et al. J. Am. Chem. Soc. 1987, 109, 5856; publication are included in this description by reference) in these conditions there was alcohol 70. Then was carried out by obtaining acid 25 in a few extra simple steps which are shown.

Scheme 12. Synthesis of allegorists 25

Reagents and conditions: (a) (i) TrocCl, pyridine, 92%; (ii) p-TsOH·H2O, 76%; (iii) LDA, tert-butyl acetate, THF, 80%; (iv) periodinane dess-Martin, 74%; (b) the catalyst Noyori (10 mol.%), MeOH/HCl, H2, 1200 lb./psi, 80%. (c) (i) TESCl, imidazole, 77%; (ii) Zn, AcOH, THF, 99%; (iii) TBSOTf, 2,6-lutidine, 82%; for the remaining stages, see Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899.

New direct and easily scalable synthesis also designed for 90 (scheme 13). The synthesis begins with the interaction of commercially available crittergetter 82 with allylbromide India. A key step in the synthesis is a specific position and stereospetsifichno dehydration of the resulting tertiary alcohol with getting 84 (with a total yield for two steps 65%). Steric regulation of this response is the result of a "bipolar"effect, which strongly dilatory electron of CF3and CO2Et best represented in the TRANS-position relative to the resulting double bond. The desired iodide 86 received in two stages of 84. Alkylation of enolate lithium 7, as described previously, iodide 86 in THF gave 88 exit 81%) and high diastereoselectivity (>25:1 de). After removal of the protection of the secondary alcohol compound 88 in three stages turned 90, as shown.

Scheme 13. Synthesis of alkyl plot 17

Reagent and conditions: (a) (i) allylbromide, In THF-water (3:1) 48°C, 85%; SOCl2, pyridine, 55°C, 77%; (b) (i) DIBAL-H, CH2Cl2from -78°C to CT 99%; (ii) I2, PPh3, imidazole, CH2Cl2, 74%; (c) (i) LHMDS, THF, -78°C to CT; (ii) HOAc-THF-H2O(3:1:1), 81% for two steps; (d) (i) AlMe3, MeONHMe, THF, 0°C to CT, 97%; (ii) MeMgBr, THF, 0°C, 53% (73% based on the extracted original substance).

In the presence of 25 and 90 in the easily implemented chemistry path 29 was clear, if you follow the Protocol, first developed by the authors in the phase of invention (A. Rivkin et al. J. Am. Chem. Soc. 2003, 125, 2899; included in this description by reference). Key metathesis reaction with snapping cycle 25 was carried out in toluene using the catalyst of Grubbs second generation Grubbs, R. H.; Miller, S. J.; Fu, G. C. Acc. Chem. Res. 1995, 28, 446; Trnka, M.; Grubbs, R. H.Acc. Chem. Res.2001, 34, 18;Alkene Metathesis in Organic ChemistryEd.: Fürstner, A.; Springer, Berlin (1998); Fürstner,A. Angew. Chem. Int.Ed. Engl.2000, 39, 3012; Schrock, R. R.Top. Were obtained. Chem.1998, 1, 1; each publication included in this description by reference). The reaction gave exclusively the TRANS isomer 48 with 71%yield. After the introduction of the rest of the thiazole by Protocol, shown in figure 14, should remove the two protective groups silila using HF-pyridine, thereby causing connection 29, which is then transformed through the restoration of 9,10-olefin 2 with high yield. Gram quantities of structurally new epothilone who received a total synthesis in the laboratory of the academic level.

Scheme 14. The final stage of synthesis of 26-CF3-(E)-9,10-degidro-dEpoB (29)

Reagents and conditions: (a) EDCI, DMAP, CH2Cl2, 25, 0°C to CT, 86% based on complex tert-butyl ether; (b) the catalyst of Grubbs, toluene, 110°C, 20 min, 71%; (c) (i) KHMDS, 101, THF, -78°C to -20°C, 70%; (ii) HF-pyridine, THF, 98%.

Experiments

Common ways: Reagents obtained from commercial suppliers were used without further purification unless otherwise stated. Methylene chloride was obtained from a dry solvent system (passed through the column, pre-filled alumina) and used without further drying. All are sensitive to the atmosphere and water of reaction was carried out in a flame dried glassware under a positive pressure of pre-purified argon gas. NMR spectra (1H and13C) were recorded on a Bruker AMX-400 MHz or Bruker Advance DRX-500 MHz, which are listed separately, using as a reference CDCl3(7,27 ppm for1H and of 77.0 ppm for13C) or CD2Cl2(5,32 ppm for1H and 53.5 ppm for13C). Infrared spectra (IR) were obtained on a spectrometer Perkin-Elmer FT-IR model 1600. Optical rotations were obtained on a digital polarimeter JASCO model DIP-370. Analytical thin-layer chromatography was carried out on plates of E. Merck silica gel 60 F254. Connect the Oia, which were not UV active, visualized by dipping the plates in a solution of para-anisaldehyde and heating. Preparative thin-layer chromatography was performed using the indicated solvent on the plate for TLC Whatman® (LK6F silica gel 60A).

Chemical substances.All epothilone synthesized own (C. R. Harris, S. J. Danishefsky, J. Org. Chem. 1999, 64, 8434; D.-S. Su et al. Angew. Chem. Int. Ed. Engl. 1997, 36, 2093; Smart, B. E. J. Fluorine Chem. 2001, 109, 3; F. Yoshimura, et al. Angew. Chem. 2003, 42, 2518; Rivkin et al. J. Org. Chem. 2002, 124, 7737; each publication included in this description by reference). Paclitaxel (Taxol®) and sulfate of vinblastine (VBL) was purchased from Sigma. All these compounds were dissolved in dimethyl sulfoxide for in vitro (except VBL, which was dissolved in saline solution). For in vivo all epothilone and paclitaxel was dissolved in the excipient cremophor/ethanol (1:1) and then diluted with saline to/in infusion within 6 hours after tail vein, using custom made minicycler (T.-C. Chou et al. Proc. Natl. Acad. Sci. USA. 2001, 98, 8113-8118; T.-C. Chou et al. Proc. Natl. Acad. Sci. U.S.A. 1998, 95, 15798-15802; each publication included in this description by reference).

Tumor and cell line.Cell lymphoblastic human leukemia CCRF-CEM received from Dr. William Beck, the University of Illinois, Chicago. Carcinoma cells human mammary gland (MX-1) and human lung carcinoma (A59) were obtained from the American type culture collection (ATCC, Rockville, MD). Resistant to paclitaxel cells A549/taxol (44-fold resistance) was created in the way described above (T.-C. Chou et al. Proc. Natl. Acad. Sci. USA. 2001, 98, 8113-8118; publication are included in this description by reference).

Animals.Nude nude mice carrying gene nu/nu, were obtained from NCI, Frederick, MD and used for all xenografts of human tumors. Used male mice 6 weeks of age or older, weighing 20-22, Drugs were injected through the tail vein for 6 hours/in infusion, using minicaster for infusion of their own making and sealed tube (T.-C. Chou et al. Proc. Natl. Acad. Sci. USA. 2001, 98, 8113-8118; publication are included in this description by reference). For/in infusion used multichannel programmable syringe pump. The usual amount during a 6-hour infusion of each drug in a mixture of cremophor/ethanol (1:1) was 100 ml in 2.0 ml of physiological solution. Tumor volume was estimated by measuring the length × width × height (or width)using a Vernier caliper. In the case of bearing tumors of nude mice in the experiment, the body weight refers to the total mass minus the mass of the tumor. All animal studies were conducted in accordance with the rules of the guide for the care and use of animals of the National institutes of health and the Protocol approved by the Institute is Kim by the Committee on care and use of animals at Memorial Sloan-Kettering Cancer Center.

Analysis of cytotoxicity.In preparation for analysis of cytotoxicity in vitro cells were cultured with an initial density of 2-5×104cells in a milliliter. They were maintained in a humid atmosphere with 5% CO2at 37°C in RPMI medium 1640 (GIBCO/BRL)containing penicillin (100 units/ml), streptomycin (100 μg/ml, GIBCO/BRL), and 5% V / V heat inactivated FBS. For growing in a monolayer of cells of solid tumors (such as A549) cytotoxicity of the drug was determined in 96-well tablets for micrometrology using the method based sulforhodamine B (P. Skehan et al. J. Natl. Cancer. Inst. 1990, 82, 1107-1112; included in this description by reference). For cells grown in suspension (such as CCRF-CEM and compared) cytotoxicity was measured in two iterations, using the method for the micro cultures hydroxide-based 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-5-carboxanilide)-2H-tetrazole (Templ) (D. A. Scudiero et al. Cancer Res. 1988, 48, 4827-4833; publication are included in this description by reference), in 96-well tablets for micrometrology. In the case of both methods, the optical density of each well was measured using a reader for microplates (Power Wave XS, Bio-Tek, Winooski, VT). Data based dose-effect obtained for 6-7 concentrations of each drug in two repetitions were analyzed on the basis of the schedule of the median effect using the computer the computer program (T.-C. Chou, M. Hayball. CalcuSyn for Windows, Multiple-drug dose effect analyzer and manual. Biosoft, Cambridge Place, Cambridge, UK (1997); included in this description by reference).

Stability epothilones in mice and in a fraction S9 human liver.The stability studies were carried out using a fully automated HPLC system, which consisted of preparation of sample Prospekt-2 (Spark Holland, Netherlands) and HPLC system Agilent 1100. Briefly, Prospekt 2 grasped cartridge for extraction of C8 and washed his acetonitrile and water. Automatic sampler Agilent mounted at 37°C, were selected 20 μl of the sample, put it on the cartridge, washed with water, then Prospekt-2 directed the flow of mobile phase through the cartridge for extraction on the analytical column Reliance Stable Bond C8 4×80 mm with protective column (MacMod, Chadds Ford, PA) and the eluent was monitored at 250 nm. The mobile phase consisted of a mixture of 53 or 65% acetonitrile/0.1% of formic acid at a flow rate of 0.4 ml/min, so that the retention time of interest compounds was approximately 6 minutes. Sample preparation consisted of adding equal volumes of plasma to PBS to a total volume of 300-400 μl, filtration and addition of 0.5-2 ál of substrate (20 mm) to achieve approximately 30-50 U at 250 nm in HPLC analysis. For combined fractions of human liver microsomes S9 (Xeno Tech, Lenex, KS), 20 ál (400 mcg) of S9 fraction was mixed with 280 μl of PBS, and then acted, campisano above. The sampling period of the sample was controlled by the automatic sampler and collected data of the peak area to compare the rate of disappearance of the parent compound.

Determination of distribution coefficient octanol-water (POW). Using the HPLC method to assess the distribution of the octanol-water. Use the HPLC system Agilent 1100 column Eclipse XDB C18 of 4.6×250 mm, mobile phase in a mixture of 60% acetonitrile/40% 25 mm potassium phosphate buffer at pH 7.4, flow rate 0.8 ml / min, and the eluent is controlled at 250 nm. Used standards are benzyl alcohol, acetophenone, benzophenone, naphthalene, diphenyl ether and dibenzyl with the famous POW 1,1; 1,7; 3,2; 4,2 and 4.8, respectively. Use sodium dichromate, to assess the origin, which is 2.5 min, and the retention times for standards is 3.9; 5,4; 10,6; 14, 18,7 19,8 min, respectively. The value of k value calculated according to the formula k=(trt-t0)/t0. Linear regression of log k versus log POW gives a straight line with r2= 0,966. This graph is used to estimate the value POW analogues epothilone.

Spectroscopic data for 29 (26 trifter-(E)-9,10-degidro-dEpoB):

[α]D25-54,6 (0,28, l3); IR (film) ν 3478, 2974, 2929, 1736, 1689, 1449, 1381, 1318, 1247, 1169, 1113, 1039, 983, 867, 736 cm-1;1H NMR (400 MHz, CDCl3) δ of 1.05 (3H, c), of 1.12 (3H, d, J=7,0 Hz)of 1.23 (3H, d, J=6,8 Hz)to 1.37 (3H, c), 2,04 (1H, userd, =3.8 Hz, -OH), a 2.12 (3H, c), 2,25 is 2.33 (1H, m), of 2.38 (1H, DD, J=15,3 and 3.0 Hz), 2,48 (1H, DD, J=15.4 9.8 Hz), 2,54-2,61 (1H, m), 2,66 was 2.76 (1H, m), 2,71 (3H, c), 2,96 (1H, DD, J=16.5 and 4.5 Hz), to 3.02 (1H, DD, J=16.3 and a 6.5 Hz), 3,11 (1H, quintet, J=6,7 Hz), 3,19 (1H, users, =OH), 3,74 (1H, users), 4,35 (1H, userd, J=9.5 Hz), 5,42 (1H, DD, J=6,2 and 4.1 Hz), the ceiling of 5.60 (1H, DDD, J=15,8, 5,6, and 4.5 Hz), to 5.66 (1H, DD, J=15,8 and 5.8 Hz), 6,24 (1H, t, J=7.2 Hz), 6,64 (1H, c), of 7.00 (1H, c);13C NMR (100 MHz, CDCl3) δ 15,1, 16,1, 17,7, 18,5, 19,3, 22,5, 28,8, 31,1, 39,6, 39,7, 45,0, 53,7, 71,4, 75,3, 76,8, 116,7, 120,2, 124,3 [kV1J(C,F) = 273,4 Hz], 127,9, 130,2 [kV3J(C,F) = 6,0 Hz], to 130.6 [kV2J(C,F) = 28.4 Hz], 132,5, 136,7, 152,0, 165,4, 170,2, 218,4; MS-HP (ESI) calculated for C27H37F3NO5S [M+H+] 544,2 found 544,1.

Example 11

In vitro studies

A typical experiment involves culturing cells (e.g., CCRF-CEM) with an initial density of 2-5×104cells in ml. support Them in a humid atmosphere with 5% CO2at 37°C in RPMI medium 1640 (GIBCO/BRL)containing penicillin (100 units/ml), streptomycin (100 μg/ml) (GIBCO/BRL) and 5% V / V heat inactivated fetal calf serum. In the case of cells that were grown in suspension (such as CCRF-CEM and compared), cytotoxicity was measured using tetragonally way to microculture hydroxide-based 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-carboxanilide)-2H-tetrazole (Templ), two duplicates in 96-well tablets for micrometrology. In the case of both optical methods is Yu density is measured using a reader for microplates (EL-340, Bio-Tek, Burlington, VT). Each series contains six or seven concentrations of the tested drugs. Data relationship dose-effect analyze graphics-based median effect.

T-cells of human CCRF-CEM cells, acute lymphoblastic leukemia, resistant to teniposide pollinia (CCRF-CEM/VM1and resistant to vinblastine pollinia (CCRF-CEM/VBL100received from W. T. Beck (University of Illinois, Chicago, I1).

In a typical experiment, which is in General described above, we offer some of the invention compounds (for example, 9,10-degidro-EpoD) showed activity in cell lines CCRF-CEM and cell lines CCRF-CEM, resistant to Taxol. Some of these compounds exhibit IC50in the range from 0,0015 to about 0,120 on cell lines CCRF-CEM. Some other compounds exhibit IC50in the range from 0,0015 to about 10.5V. Some of these compounds also exhibit IC50in the range from to 0.011 to about 0.80 to line resistant cells CCRF-CEM/Taxol and some other compounds exhibit IC50in the range of from about 0,011 to 13.0 μm. In some embodiments, 26F-EpoD shows activity within 0,0015 μm on the cell lines CCRF-CEM and within 0,011 μm lines resistant cells CCRF-CEM/Taxol (11).

Example 12

In vivo studies

Usually xenografts tumors used Nude mice nude gene bearing nu/nu. Outbred mice, derived from Swiss, were obtained from Charles River Laboratories. For most experiments used male mice 8 weeks of age or older, weighing 22, the Drug was injected through the tail vein through a 6-hour/in infusion. Each mouse was holding vessel for immobilization of polypropylene Falcon tube with holes for injection of a medicinal product. Tumor volume was estimated by measuring the length × width × height (or width)using a Vernier caliper. For/in infusion used multichannel programmable syringe pump Harvard PHD2000 (Harvard Apparatus). All animal studies were carried out in accordance with the rules of the "Guide for the care and use of animals" of the National Institute of health and the Protocol approved by the Institutional Committee for care and use of animals at Memorial Sloan-Kettering Cancer Center. In accordance with the regulations of the Committee in relation to the humane treatment of bearing tumors of animals mice were subjected to euthanasia when the tumor reached >10% of their total body weight.

As shown in Fig, 9,10-degidro-EpoB tested in nude mice bearing carcinoma human breast MX-1. In General 9,10-degidro-EpoB was prepared as follows: 9,10-degidro-EpoB was dissolved in ethanol and added cremophor (1:1) at a concentration of 20 mg/ml of This solution was diluted Phi is biologicheskii solution for/in infusion. The diluted solution was used for/in infusion over one hour. Then measured the tumor size and body weight, using a dose of 10 mg/kg, 20 mg/kg and 30 mg/kg for 15 days. Tumor size and body weight were also measured using the schema dosage of 0.4 mg/kg Q3Dx2, 0.5 mg/kg Q3Dx2 and 0.6 mg/kg Q3Dx5 (see Fig, 34, 55 and 56). Used the scheme of dose every third day, to reduce toxicity. Other therapeutic studies 9,10-degidro-EpoB shown in Fig and 71 (CCRF-CEM/Taxol Q3Dx5) and Fig and 24 (HCT-116, Q2Dx7).

The compound 9,10-degidro-12,13-desoxyepothilone B (ISO-490-epothilone) is three times more effective than dEpoB. It is shown that 9,10-degidro-12,13-desoxyepothilone D inhibits tumor growth after two to three infusions at doses of 10 mg/kg or 20 mg/kg, each of which was injected through the day. Better results were obtained in mice using doses of 30 mg/kg 9,10-degidro-12,13-desoxyepothilone B using 6-hour infusion/in through the day. Also it is shown that in the case of 9,10-degidro-dEpoB at a dose of 5 mg/kg, Q3Dx9, 6 hours/in infusion results in disappearance of tumors in nude mice bearing xenograft MX-1, without loss of mice and with only moderate loss of body weight (Fig and 75). Apparently, this is done by introducing analogues epothilone every third day, to reduce toxicity (see Fig and 54). In conclusion, 9,10-degidro-12,13-desoxyepothilone B is reavley reduced toxicity compared to other epothilone, higher efficiency in delayed tumor growth and increased stability in serum. Other therapeutic studies shown in Fig and 18 (HCT-116, Q2Dx5 and Q3Dx5); Fig and 20 (A549/Taxol, Q3Dx7); and Fig and 22 (A549/Taxol, Q2Dx7).

9,10-degidro-Epo B in the introduction of every third day 9-11 times 6-hour/in infusion at a dose of 0.4-0.6 mg/kg resulted in reduction and disappearance of tumors in nude mice with implanted xenografts of breast carcinoma human MX-1 (Fig and 69). Introduction through day 8 doses resulted in suppression of tumor growth, but not the tumor to shrink. When 9,10-degidro-Epo B was injected through the day of 9 doses, planted the tumor continued moderate decline from the second to the eighth day, but the body weight was recovering very slowly from 76% to 82% of control during the same time period. On the tenth day of one quarter of the tumor was gone. When injected dose of 0.6 mg/kg 9,10-degidro-EpoB, Q2Wx6, 6-hour infusion of nude mice with xenografts HCT-116, four of four mice died of toxicity within three days after six doses. 9,10-degidro-EpoB stopped tumor growth CCRF-CEM/Taxol using 0.6 mg/kg, scheme Q3Dx5, x2 (Fig and 71).

26 Trifter-9,10-degidro-12,13-desoxyepothilone B (F3-deH-dEpoB), as shown in the figures, has a curative effect in doses of 20mg/kg and 30 mg/kg, Q2Dx6, 6-hour infusion, model nude, implanted by xenografts of breast carcinoma human MX-1. The data also indicate that 30 mg/kg, Q2Dx6 is approximately the maximum tolerated dose. At a dose of 20 mg/kg, Q2Dx6, 6-hour infusion, 26 trifter-9,10-degidro-12,13-desoxyepothilone B led to the reduction and disappearance of the tumor in four out of four nude mice with xenografts of breast carcinoma human MX-1. Not seen the re-appearance of the tumor on the 20th day after the end of treatment. On the 27th day after the end of treatment re-appeared 2/4. Had no further recurrence of tumors within 28-64 days after stopping treatment. In comparison, in the case of dEpoB at a dose of 30 mg/kg was achieved by the disappearance of the tumor in the same mouse model for seven of seven mice; however, the tumor re-appeared in 2 out of five mice on day 8 after stopping treatment. Introduction 26 trifter-9,10-degidro-12,13-desoxyepothilone B at a dose of 20 mg/kg, Q2Dx6, 6-hour/in infusion, led to the temporary reduction of body weight of the mice to 26%. This decrease in body weight did not lead to death, indicating the absence of severe toxicity in relation to vital organs. Two days after the last treatment, the body weight started to recover. At 16 days after treatment body weight was restored to 109% of control to ensure that the witness is Alstom about the toxicity, if any, is fully reversible. In comparison dEpoB, administered at a dose of 30 mg/kg, resulted in a 31% decrease in body weight without mortality.

In that case, when 26 trifter-9,10-degidro-12,13-desoxyepothilone B was administered at a dose of 30 mg/kg, Q2Dx6, 6-hour/in infusion, the disappearance of the tumor was 2-3 days earlier than at the dose of 20 mg/kg In the case of this higher dose body weight was decreased by 27% and was maintained for 4 days, without causing lethality, which confirms the absence of severe toxicity in relation to vital organs. Four days after the last treatment at a dose of 30 mg/kg body weight began to recover. On the 16th day after treatment, body weight was restored to 98% of control to ensure that once again confirms the reversibility of toxicity. Treatment 26 trifter-9,10-degidro-dEpoB at a dose of 20 mg/kg and 30 mg/kg resulted in complete disappearance of the tumors, and with the dose of 30 mg/kg were observed relapse after 62 days. The disappearance of the tumors was also achieved in the case of 10 mg/kg with the introduction of 9 doses, when given three additional doses (Fig). At the dose of 26 trifter-9,10-degidro-dEpoB at a dose of 10 mg/kg was observed only a small loss of body weight (Fig). Was not observed further loss of body weight in long-term treatment.

On Fig summarized the effects of 26-F3-9,10-deH-dEpo B (and other epothilones) against xenograft X-1, A. At low dose; B. Against large tumors; against xenograft lung carcinoma A549, C; and against resistant to Taxol xenograft lung carcinoma A549/Taxol, D.

On Fig lists, indicating the efficiency of in vitro C-21 modified epothilone against CCRF-CEM, CCRF-CEM/VBL and CCRF-CEM/Taxol.

On Fig shown therapeutic effects of 26-F3-9,10-deH-dEpoB (15 mg/kg and 30 mg/kg) and Taxol (20 mg/kg) Q2Dx8, 6-hour/in infusion, against xenograft T-cell lymphoblastic human leukemia CCRF-CEM. A similar decrease in body weight was observed in all three treatment groups (Fig).

In the treatment of xenograft CCRF-CEM/Taxol (resistant to Taxol) 26-F3-9,10-deH-dEpo B at a dose of 15 mg/kg was achieved disappearance 1/3 of tumors, and in a dose of 30 mg/kg was achieved disappearance 3/4 tumors. The same treatment with Taxol at a dose of 20 mg/kg were given only partial suppression of tumor growth and failed to achieve reduction of the tumor (Fig). Changes in body mass during the experiment shown in Fig.

In the treatment of xenograft carcinoma of the colon of human HCT-116 26-F3-9,10-deH-dEpo B (20 mg/kg) reached similar efficiency as in the case of Taxol (20 mg/kg). However, F3the deH-dEpo B at a dose of 30 mg/kg exerted a therapeutic effect is better with the disappearance 2/4 tumors after 5 doses (Fig). Changes in body mass during Dan is on the experiment is shown in Fig.

Therapeutic effects F3-9,10-degidro-dEpoF against xenografts MX-1 at different doses (5-30 mg/kg) in the case of a 6-hour/in infusion and/in injection shown in Fig and 77.

Conclusion.9,10-degidro-, 26-Cryptor or both versions of dEpoB lead to a 1.5-5-fold increase in cytotoxicity in vitro and 2-5-fold increased half-life in plasma of mice in vitro. Using the model xenografts solid human tumors in nude mice and using the methodology Q2Dx5~9, 6 hours/in infusion via tail vein at a maximum-tolerated doses evaluated the antitumor efficacy and toxicity of 9,10-dehydroemetine. The ability to achieve complete suppression of tumor growth, reduction and disappearance of tumors promoted further research in order to determine the frequency of relapses and the indicator of treatment efficacy after discontinuation of treatment. 9,10-degidro-EpoB, the most effective in vitro known aptilon, although it was highly effective, but showed the narrow limits of therapeutic safety in vivo. 9,10-degidro-dEpoB at a dose of 4 mg/kg, 9,10-degidro-EpoB at a dose of 0.4 mg/kg and 21-hydroxy-9,10-degidro-dEpoB at a dose of 3 mg/kg - all strongly inhibited tumor growth over a long period of time and have achieved some reduction of tumors, and sometimes reached the disappearance of the tumors. dEpoB at a dose of 30 mg/kg, 26-rifter-9,10-degidro-dEpoB at a dose of 20 mg/kg) and paclitaxel at a dose of 20 mg/kg all showed strong suppression of tumor growth and achieved reduction and disappearance of tumor xenografts of breast carcinoma human MX-1 in all tested mice. 26 trifter-9,10-degidro-dEpoB compared with dEpoB or paclitaxel was achieved long-term recovery without relapse and equally demonstrated rapid recovery of body weight to control the level before treatment.

Example 13

The synthesis of analogues cyclopropylacetylene

1. The compound of the formula:

or its pharmaceutically acceptable salt,
in which R1means hydrogen or C1-6alkyl;
R2means isooxazolyl group, substituted C1-6by alkyl;
RBmeans-CF3, -CHF2, -CH2F, or C1-6alkyl.

2. The compound according to claim 1, in which R1means C1-6alkyl.

3. The compound according to claim 2, in which R1means methyl.

4. The compound according to claim 3 in which RBmeans-CF3.

5. The compound according to claim 3 in which RBmeans methyl.

6. The compound according to claim 1, in which R2replaced by stands.

7. Pharmaceutical composition for treating cancer containing the compound according to any one of claims 1 to 6 and a pharmaceutically acceptable excipient.

8. The pharmaceutical composition according to claim 7, additionally with the holding cremophor.

9. The pharmaceutical composition according to claim 7, further containing cremophor and ethanol.

10. The pharmaceutical composition according to claim 7, where the connection is suspended in a mixture of cremophor/ethanol in a ratio of 1:1.

11. The pharmaceutical composition according to claim 7, further containing additional cytotoxic agent.

12. Pharmaceutical composition for treating cancer containing a therapeutically effective amount of a compound according to any one of claims 1 to 6, or its pharmaceutically acceptable salt; and a pharmaceutically acceptable carrier or diluent,
moreover, a therapeutically effective amount of compound is an amount sufficient to deliver about 0.001 mg to about 40 mg of compound per kg weight of the subject.

13. The use of compounds according to any one of claims 1 to 6 in the manufacture of a medicinal product for the treatment of cancer.

14. Use item 13, where a therapeutically effective amount of compound is an amount sufficient to deliver about 0.001 mg to about 40 mg of compound per kg weight of the subject.

15. Use item 13, where a therapeutically effective amount of compound is an amount sufficient to deliver about 0.1 mg to about 25 mg of compound per kg weight of the subject.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new substituted heteroaryl derivatives of general formula I: , wherein: A means N, CR7-10, with A at the most twice meaning N; W means O, S or NR4, the values B, C, R7-10 are presented in clause 1 of the patent claim. The method for preparing the compound I is described.

EFFECT: compounds show analgesic activity that enables using them for a variety of diseases, especially acute pain, neuropathic, chronic or inflammatory pain.

16 cl, 2 tbl, 307 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of general formula (I),

, where groups and radicals R1, R2 independently denote H, C1-8-alkyl or C3-7-cycloalkyl, where the alkyl or cycloalkyl group can be mono- or poly-substituted with identical or different groups R11; or R2 denotes a -CH2- or -CH2-CH2- bridge which is bonded with a group Y, and R1 is as defined above, or denotes a group selected from C1-4-alkyl-CO-, C1-4-alkyl-O-CO-, (C1-4-alkyl)NH-CO- or (C1-4-alkyl)2N-CO-, where the alkyl groups can be mono- or polyfluorinated; or R1 and R2 form an alkylene bridge such that R1R2N- denotes a group selected from: azetidine, pyrrolidine, piperdine, azepan, 2,5-dihydro-1H-pyrrole, 1,2,3,6-tetrahydropyridine, 2,3,4,7-tetrahydro-1H-azepine, 2,3,6,7-tetrahydro-1H-azepine, piperazine, in which the free amino group is substituted with R13, piperidin-4-one, morpholine, thiomorpholine, 4-C1-4-alkoxy iminopiperidin-1-yl and 4-hydroxy iminopiperidin-1-yl. Wherein, when R1 and R2 form an alkylene bridge, one or more H atoms in the alkylene bridge can be substituted with identical or different groups R14, and X denotes a C1-3-alkylene bridge which can contain one, two or three identical or different C1-3-alkyl substitutes; and Y denotes a group of subformula selected from: and , where the group can be mono-substituted with a substitute R20; Z denotes -CH2-CH2- or -C(=O)-CH2-; U, V both denote CH, one of groups U, V denotes N, and the other of U, V denotes CH, where CH can be substituted with L; and L independently denotes halogen, cyano or C1-3-alkyl; and k equals 0, 1 or 2; W is selected from a group consisting of -CH2-O- and -O-CH2-; B is selected from a group consisting of phenyl, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furyl, thiophenyl and thiazolyl; each of which can be mono- or poly-substituted with identical or different substitutes R20; radicals R11, R13, R14, R20 assume values given in claim 1. The invention also relates to a pharmaceutical composition containing at least one compound of formula I and having action on MCH receptor.

EFFECT: disclosed pharmaceutical compositions are useful in treating metabolic disorders or eating disorders, especially obesity, bulimia, anorexia, hyperphagia and diabetes.

FIELD: chemistry.

SUBSTANCE: invention relates to novel substituted pyrimidine derivatives, having HIV replication inhibiting properties, or pharmaceutically acceptable salts thereof. In formula (1): R1 denotes hydrogen; R2 and R3 independently denote hydrogen; R7 and R8 denote C1-6alkyl; R4 denotes cyano; R9 denotes C1-6alkyl optionally substituted with cyano, C2-6alkenyl substituted with cyano, C2-6alkynyl optionally substituted with cyano; R5 denotes C1-6alkyl optionally substituted with Ar or Het; C2-6alkenyl optionally substituted with Ar or Het; C2-6alkynyl optionally substituted with Ar or Het; C3-7cycloalkyl; Ar; Het; R6 denotes H, Het; Y denotes -OR11, -NR12R13; R11 denotes hydrogen or C1-6alkyl optionally substituted with hydroxy, C1-6alkoxy or pyridyl; R12 denotes hydrogen or C1-6alkyl; R13 denotes hydrogen or C1-6alkyl; or R12 and R13 together with a nitrogen atom, which is substituted by said two substitutes, form a morpholinyl; imidazolyl; X denotes -NR1-; Het denotes 5- or 6-member completely saturated ring, where one or two ring members are heteroatoms, each independently selected from nitrogen and sulphur, and where the rest of the ring members are carbon atoms; and where any member of the heterocycle with a nitrogen heteroatom can optionally be substituted with C1-6alkyl; where the 5- or 6-member ring can optionally be annelated with a benzene or thiophene ring; each aryl independently denotes phenyl or phenyl substituted with one substitute selected from C1-6alkoxy.

EFFECT: high efficiency of using said compounds.

7 cl, 4 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing mono-(di-, tetra-)methyl-1,2-bis-(1,3,5-dithiazinan-5-yl)ethanes of general formula (1-3): , (1) R1=CH3, R2, R3, R4=H (2) R1, R3=CH3, R2, R4=H (3) R1, R2, R3, R4=CH3, which involves reaction of hydrogen sulphide-saturated aqueous solution of (37%) formaldehyde and acetaldehyde with 1,2-diaminoethane with molar ratio of initial reagents 1,2-diaminoethane: formaldehyde: acetaldehyde: hydrogen sulphide equal to 10:50:10:40 to obtain (1), 10:40:20:40 to obtain (2), 10:20:40:40 to obtain (3), at temperature 40°C and atmospheric pressure for 2.5-3.5 hours.

EFFECT: method of obtaining novel compounds which can be used as selective sorbents and extraction agents of precious metals, agents for protecting leather, fur and textile from biodeterioration, biologically active substances with respect to different microorganisms and sulphate-reducing bacteria.

1 cl, 1 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing 1,2-bis-(2,4,6-trialkyl-1,3,5-dithiazinan-5-yl)ethanes of general formula (1): . The method involves reaction of a hydrogen sulphide-saturated aldehyde of formula RCHO, where R=CH3, C2H5, n-C3H7, n-C4H9, n-C5H11, with 1,2-diaminoethane with molar ratio of initial reagents 1,2-diaminoethane: aldehyde: hydrogen sulphide equal to 10:60:40 at temperature 40°C and atmospheric pressure for 2.5-3.5 hours.

EFFECT: method of obtaining novel compounds which can be used as selective sorbents and extraction agents of precious metals, agents for protecting leather, fur and textile from biodeterioration, biologically active substances with respect to different microorganisms and sulphate-reducing bacteria.

1 cl, 1 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing 2(3H)-benzothiazolone. The method is realised by boiling bis-(2,2'-dimethoxycarboxamido)-phenyl disulphide with zinc dust for 7,5 hours in glacial acetic acid, followed by separating zinc salts, diluting the filtrate with ice water, filtering the crystalline product and washing said product with diluted HCl (1:1) and water and then drying on air and recrystallising from ethanol. The invention also relates to a method of producing 3-2-[2-oxo-1,3-benzothiazol-3(2H)-yl]ethyl-1,3-benzothiazol-2(3H)-one. The method is realised by boiling 2(3H)-benzothiazolone with 1,2-dibromoethane in acetone in the presence of K2CO3. Water is then added and the mixture is treated with diethyl ether, followed by washing the organic phase with 10% aqueous NaOH solution and water, drying with anhydrous potassium carbonate, removing the solvent and recrystallising from methanol. The invention relates to a method of producing 3-(2-chloroacetyl)-1,3-benzothiazol-2(3H)-one. The method is realised by boiling 2(3H)-benzothiazolone in anhydrous benzene with chloroacetyl chloride for 10 hours, then washing the cooled reaction mass with 2% aqueous NaOH solution, extracting with benzene, washing the merged extracts with water, drying with anhydrous calcium chloride and removing the solvent, followed by recrystallisation from chloroform.

EFFECT: improved method of producing 2(3H)-benzothiazolone derivatives.

3 cl, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to tetrahydroquinoline derivatives of formula (I), where values of C3-C4, R2, R3, R4, R5, L1, L2, Y and X are given in claim 1, as muscarinic receptor agonists; compositions containing said compounds; methods of inhibiting muscarinic receptor activity using said compounds; methods of treating diseased conditions associated with the muscarinic receptor using said compounds, and methods of identifying a subject suitable for treatment using said compounds.

EFFECT: improved properties of compounds.

22 cl, 1 tbl, 3 ex, 3 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to novel carbostyril compounds of general formula (1) or salts thereof with common pharmaceutically acceptable acids or pharmaceutically acceptable basic compounds, having activity on promotion of TFF2 production, a pharmaceutical composition based on said compounds, an agent based on disclosed compounds used in case of a disorder where up-regulation of TFF has a prophylactic and/or therapeutic effect, use of disclosed compounds to prepare said agent and a method of producing disclosed compounds. The invention also relates to novel specific carbostyril compounds or salts thereof with common pharmaceutically acceptable acids or pharmaceutically acceptable basic compounds. In structural formula (1), A is a direct bond, a lower alkylene group or lower alkylidene group, X is an oxygen or sulphur atom, the bond between positions 3 and 4 of the carbostyril backbone is a single bond or a double bond, R4 and R5 each denotes a hydrogen atom provided that, when the bond between positions 3 and 4 of the carbostyril backbone is a double bond, R4 and R5 can instead be bonded to each other in form of a -CH=CH-CH=CH- group, and R1, R2 and R3 assume values given in the claims.

EFFECT: high efficiency of compositions based on said compounds.

32 cl, 23 dwg, 184 tbl, 1535 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of general structural formula:

or to pharmaceutically acceptable salts, where Z denotes -O- or -CH2- or -CH2-CH2-; X1 denotes a covalent bond or -O-; Y1 denotes a covalent bond or C1-C10 alkylene, provided that Y1 is a covalent bond only when X1 denotes a covalent bond; R1 denotes a) (C3-C7)cycloalkyl or b) phenyl or heteroaryl, which is a monovalent heteroatomatic monocyclic radical ring containing 1-2 heteroatoms, independently selected from nitrogen and sulphur, possibly substituted with 1-3 groups, independently selected from fluorine, chlorine, bromine, (C1-C6)alkyl or (C1-C6)-alkoxy; R2 denotes -OC(O)(NH2), -OC(O)(NHR9), -NHC(O)OR9, -C(O)R9, -C(O)(NH2), -C(O)(NHR9) or -NHC(O)H, where R9 denotes a linear or branched C1-C5 alkyl or a linear or branched (C1-C5)alkoxyalkyl; R3 denotes H, C1-C5 alkyl, -NHC(O)R10 or OH, where R10 denotes C1-C3 alkyl, provided that when R3 denotes -OH, X1 is not O and R2-Y1-X1 is not -OC(O)(NH2), -OC(O)(NHR9), -NHC(O)OR9 or -NHC(O)H; -Q denotes

, where N and N are bonded by bonds denoted by a wavy line; R4 denotes H; R5 and R6 independently denote: a) H, (C1-C10)alkyl, (C4-C10)cycloalkylalkyl, hydroxylated (C4-C10)cycloalkylalkyl, halo(C4-C10)cycloalkylalkyl, (C1-C2)alkyl(C4-C10)cycloakylalkyl, (C4-C10)bicycloalkyl(C1-C3)alkyl, (C1-C5)alkoxy(C1-C5)alkyl; or a saturated heterocyclyl(C1-C3)alkyl, where the saturated heterocyclic ring is selected from 5-, 6- or 7-member heterocyclic rings which contain 1 heteroatom independently selected from N and O; or b) phenyl(C1-C2)alkyl, phenoxymethyl, each of which is possibly with 1-3 groups independently selected from fluorine, chlorine, (C1-C3)alkyl, (C1-C3)alkoxy; provided that both R5 and R6 are not H; G denotes NH2 or NHR7; R7 denotes (C1-C6)alkyl; or R5 and R7 together denote -CH2, -(CH2)2 or -(CH2)3, possibly substituted with 1-2 groups independently selected from (C1-C8)-alkyl, (C3-C6)cycloalkyl, (C3-C6)cycloalkyl(C1-C2)alkyl or (C1-C8)alkoxy. The invention also relates to compounds selected from the group, pharmaceutical compositions, a method for antagonising one or more aspartate proteases, as well as methods of treating aspartate protease-mediated disorders.

EFFECT: obtaining novel biologically active compounds having activity towards rennin.

35 cl, 33 ex, 4 dwg

FIELD: medicine.

SUBSTANCE: compounds can be used for treating neurological conditions, more specifically for treating neurodegenerative conditions, such as Alzheimer's disease. In a compound of formula I R2 represents H or CH2NR1R4 where R1 and R4 are independently selected from H, unsubstituted C1-6alkyl, substituted or unsubstituted C3-6 cycloalkyl, R3 represents H; substituted or unsubstituted C1-4alkyl; substituted or unsubstituted C2-4alkenyl; substituted or unsubstituted 6-members aryl condensed or uncondensed with substituted or unsubstituted 6-members aryl or 5-6-members heteroaryl, containing 1-2 nitrogen atoms in a cycle; substituted or unsubstituted saturated or unsaturated 5 or 6-members N-containing heterocycle which can additionally contain nitrogen, oxygen or the sulphur atom condensed or ucondensed with substituted or unsubstituted 6-members aryl or 5-6-members heteroaryl containing nitrogen in a cycle; (CH2)nR6 where n is an integer from 1 to 6, and the values of R6 and the values of other radicals are specified in the patent claim.

EFFECT: increased antiamyloidogenic action.

20 cl, 20 tbl, 6 dwg, 7 ex

FIELD: chemistry.

SUBSTANCE: in general formula 1 , a cyclic ring can be selectively formed; each of R1 and R2 is independently selected from a group consisting of hydrogen, with a straight or branched alkyl chain having 1-6 carbon atoms and phenetyl or R1 and R2 together form a 5- or 6-member heterocyclic ring or R1 and R2 together with Ar1 form a bicyclic ring; Ar1 is selected from a group consisting of furanyl, thionyl, methylene dioxyphenyl and phenyl, which can be substituted with at least one identical or different substitutes selected from a group consisting of hydrogen, with a straight or branched alkyl chain having 1-6 carbon atoms, a halogen such as F, O and Br, with a straight or branched alkoxy chain, having 1-6 carbon atoms, nitro and trifluoromethyl; Z is hydrogen or fluorine, or taken together with Ar1 forms a bicyclic ring; Ar2 is selected from a group consisting of phenyl, methylene dioxyphenyl, pyridine pyrimidine, naphthyl, bis(fluorophenyl)methyl and quinoxaline, which can be substituted with at least one identical or different substitutes selected from a group consisting of hydrogen, with a straight or branched alkyl chain, having 1-6 carbon atoms, hydroxy, halogen, with a straight or branched alkoxy chain having 1-6 carbon atoms, nitro, acetyl, tert-butyl acetate, trifluoromethyl, amino and acetate; n is equal to 1 or 2; m is an integer from 0 to 2.

EFFECT: improved method.

26 cl, 2 dwg, 2 tbl, 87 ex

FIELD: chemistry.

SUBSTANCE: invention is a 6-10-member aryl selected from phenyl, naphthyl, tetrahydronaphthalenyl, indanyl or a 6-member heteroaryl containing 1-2 N atoms, selected from pyridyl or pyrimidinyl, where the aryl and heteroaryl groups can be unsubstituted or substituted with 1-3 substitutes selected from a group consisting of C3-6-cycloalkyl, phenyl, phenyloxy, benzyl, benzyloxy, halogen atom, C1-7-alkyl, C1-7-alkoxy, oxazolyl, piperidin-1-yl, or C1-7-alkyl, substituted with a halogen atom, or represents phenyl, where at least one hydrogen atom is substituted with deuterium or tritium; R2 is a hydrogen atom, C1-7-alkyl or is benzyl, unsubstituted or substituted C1-7-alkoxy or halogen atom; or R1 and R2 together with a N atom with which they are bonded form 2,3-dihydroindol-1-yl or 3,4-dihydro-1quinolin-1-yl. The invention also relates to a method of producing compounds of formula and to a pharmaceutical composition having high affinity for the TAAR1 receptor.

EFFECT: compounds of formula (I), having high affinity for the TAAR1 receptor.

29 cl, 4 dwg, 1 tbl, 183 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new substituted heteroaryl derivatives of general formula I: , wherein: A means N, CR7-10, with A at the most twice meaning N; W means O, S or NR4, the values B, C, R7-10 are presented in clause 1 of the patent claim. The method for preparing the compound I is described.

EFFECT: compounds show analgesic activity that enables using them for a variety of diseases, especially acute pain, neuropathic, chronic or inflammatory pain.

16 cl, 2 tbl, 307 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new imidazole derivatives of formula I wherein R1 represents a hydrogen atom or C1-7alkyl; R2 represents C1-7alkyl; R3 represents C1-7alkyl, C1-7alkoxy, phenyloxy, benzyloxy, a halogen atom or C1-7alkyl substituted by a halogen atom; R4 represents a hydrogen atom or C1-7alkyl; X represents -CH2-, -CHR2 - or -O; Y represents -CH2-, -CH2CH2- or a bond; provided X represents -O-, Y represents -CH2-; Z represents -CH2- or -CHR2-; provided R2 is found twice, simultaneously for X and Z which are CHR2 , then R2 can be identical alkyls or different; n has the value 0, 1 or 2; provided n has the value 2, R3 can be identical or different; and its pharmaceutically acceptable acid addition salts, except for the following compounds: 1-(1H-imidazol-4-ylmethyl)-1,2,3,4-tetrahydro-quinoline and 1-(3H-imidazol-4-ylmethyl)-2,3-dihydro-1H-indole. Also, the invention refers to a method for preparing the compounds of formula I, to a drug based on the compound of formula I and applying the compound of formula I in preparing the drug.

EFFECT: there are prepared new imidazole derivatives effective in treating such pathological conditions, as bipolar disorder, stress-induced disorder, psychotic disorders, schizophrenia, neurological conditions, Parkinson's disease, neurodegenerative disorders, Alzheimer's disease.

13 cl, 61 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a compound of formula (II)

or to its pharmaceutically acceptable salt wherein the ring A represents a group presented by formula 2 R1 represents hydrogen or C1-6alkyl; R2 represents -SR5, halogen, halogenated C1-6alkyl, etc., R3 represents a group presented by formula: -CH=CH-C(RaRb)-Rc-Rd, or a group presented by formula: -(CReRf)m-C(RaRb)-Rc-Rd wherein radicals and symbols have the values presented in the patent claim, R4 represents -OR6, -CONR7R8, -NR9CONR7R8, -(CR10R11)POH, -(CR10R11)pOCONR7R8, -NR9COR12, -(CR10R11)pNR9COR12, -C(=O)NR9OR12, -CONR9CONR7R8, -CN, halogen or NR9(C=O)OR12; R5 represents C1-6alkyl; R6 represents -CONR7R8; each of R7 and R8 independently represents hydrogen or etc., R10 and R11 independently represents hydrogen; R12 represents C1-6alkyl; each of m and p independently represents an integer 1 to 3. This compound is applicable as a type 1 11β-hydroxysteroiddehydrogenase inhibitor.

EFFECT: invention also refers to single compounds, pharmaceutical compositions on the basis of the declared compounds, to a method for preventing and treating diabetes and the use of the compound of formula (II).

21 cl, 289 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a novel clathrate complex of β-cyclodextrin with 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine of formula : with molar ratio 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine: β-cyclodextrin from 1:1 to 1:10, synthesis method and use thereof as an antiviral agent for treating influenza. The disclosed method involves mixing solutions of β-cyclodextrin and 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine in molar ratio from 1:1 to 1:10 while stirring and heating to temperature not higher than 70°C and then maintaining said conditions until a homogeneous solution is obtained and extraction of the obtained complex.

EFFECT: clathrate complex is a novel effective anti-influenza virus agent which is obtained using a novel efficient method.

13 cl, 2 ex, 3 tbl, 11 dwg

FIELD: chemistry.

SUBSTANCE: present invention relates to a method for synthesis of 2-substituted azole

compounds of formula , (a) reaction of an aldehyde of formula

with an azole of formula in the presence of a carbonylating agent of formula to obtain an oxazolidone of formula , reaction of the oxazolidone of formula (Ia) so as to perform hydrolysis of a triarylmethyl group, splitting the O-(C=Q) bond and opening the oxazolidone, followed by reaction of the obtained intermediate compound with Prot-Z, where Prot-Z is an agent which protects an amino group, to obtain an azole-containing intermediate compound of formula (lb) and and oxidation of the intermediate compound of formula (Ib) to obtain a 2-substituted azole derivative of formula (I). The invention also relates to azole compounds of formulae (I), (la), (lb), (Ic) and (II).

EFFECT: novel method of obtaining azoles of formula (I), as well as obtaining novel compounds of formulae (I), (la), (lb), (Ic) and (II), having useful biological properties.

41 cl, 5 tbl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula I:

or pharmaceutically acceptable salts thereof, in which Q is a divalent or trivalent radical selected from C6-10aryl and heteroaryl; where said aryl or heteroaryl in Q is optionally substituted up to 3 times with radicals independently selected from halogen, C1-6 alkyl, C1-6 alkyl substituted with halogen, C1-6 alkoxy group, C1-6 alkoxy group substituted with halogen, -C(O)R20 and -C(O)OR20; where R20 is selected from hydrogen and C1-6 alkyl; and where optionally, the carbon atom neighbouring W2 can be bonded through CR31 or O with a carbon atom of Q to form a 5-member ring condensed with A and Q rings; where R31 is selected from hydrogen and C1-6 alkyl; W1 and W2 are independently selected from CR21 and N; where R21 is selected from hydrogen and -C(O)OR25; where R25 denotes hydrogen; ring A can contain up to 2 carbon ring atoms substituted with a group selected from -C(O)-, -C(S)- and -C(=NOR30)- and can be partially unsaturated and contain up to 2 double bonds; where R30 denotes hydrogen ; L is selected from C1-6alkylene, C2-6alkenylene, -OC(O)(CH2)n-, -NR26(CH2)n- and -O(CH2)n-; where R26 is selected from hydrogen and C1-6 alkyl; where n is selected from 0, 1, 2, 3 and 4; q is selected from 0 and 1; t1, t2, t3 and t4 are each independently selected from 0, 1 and 2; R1 is selected from -X1S(O)0-2X2R6a, -X1S(O)0-2X2OR6a, -X1S(O)0-2X2C(O)R6a, -X1S(O)0-2X2C(O)OR6a, -X1S(O)0-2X2OC(O)R6a and -X1S(O)0-2NR6aR6b; where X1 is selected from a bond, O, NR7a and C1-4alkylene; where R7a is selected from hydrogen and C1-6alkyl; X2 is selected from a bond and C1-6alkylene; R6a is selected from hydrogen, cyanogroup, halogen, C1-6alkyl, C2-6alkenyl, C6-10aryl, heteroaryl, heterocycloalkyl and C3-8cycloalkyl; where said aryl, heteroaryl, cycloalkyl and heterocycloalkyl in R6a is optionally substituted with 1-3 radicals independently selected from hydroxy group, halogen, C1-6alkyl, C1-6alkyl substituted with a cyano group, C1-6alkoxy group and C6-10aryl-C1-4alkoxy group; and R6b is selected from hydrogen and C1-6alkyl; R3 is selected from hydrogen, halogen, hydroxy group, C1-6alkyl, C1-6alkyl substituted with halogen, C1-6alkyl substituted with a hydroxy group, C1-6alkoxy group, C1-6alkoxy group substituted with halogen, -C(O)R23 and -C(O)OR23; where R23 is selected from hydrogen and C1-6alkyl; R4 is selected from R8 and -C(O)OR8; where R8 is selected from C1-6alkyl, heteroaryl, C3-8cycloalkyl and heterocycloalkyl; where said heteroaryl, cycloalkyl or heterocycloalkyl in R8 is optionally substituted with 1-3 radicals independently selected from halogen, C1-6alkyl, C3-8cycloalkyl and C1-6alkyl substituted with halogen; R5 is selected from hydrogen, C1-6alkyl substituted with a hydroxy group, and a C1-6alkoxy group; heteroaryl denotes a monocyclic or condensed bicyclic aromatic ring complex containing 5-9 carbon atoms in the ring, where one or more ring members are heteroatoms selected from nitrogen, oxygen and sulphur, and heterocycloalkyl denotes a saturated monocyclic 4-6-member ring in which one or more said carbon atoms in the ring are substituted with a group selected from -O-, -S- and -NR-, where R denotes a bond, hydrogen or C1-6alkyl. The invention also relates to pharmaceutical compositions containing said compounds, and methods of using said compounds to treat or prevent diseases or disorders associated with GPR119 activity, such as obesity, type 1 diabetes, type 2 sugar diabetes, hyperlipidemia, type 1 autopathic diabetes, latent autoimmune diabetes in adults, type 2 early diabetes, child atypical diabetes, adult diabetes in children, malnutrition-associated diabetes and diabetes in pregnant women.

EFFECT: improved properties of compounds.

27 cl

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula

,

and pharmaceutically acceptable salts and solvates thereof, in which R1 is an optionally substituted alkyl or similar, R2 is a group of formula: -Y-R5, where Y is -O- or S; R5 is a substituted alkyl (the substitute is an optionally substituted cycloalkyl or similar), a branched alkyl or similar; R4 is hydrogen or C1-10 alkyl; R3 is a group of formula: -C(=O)-Z-R6, where Z is -NR7- or -NR7-W-; R6 is an optionally substituted cycloalkyl or similar; R7 is hydrogen or C1-10 alkyl, W is C1-10 alkylene; X is =N- provided that a compound in which R2 is 2-(4-morpholino)ethoxy, 2-, 3- or 4-pyridylmethoxy, 1-methylpiperidinyl-2-methoxy, benzyloxy or 4-substituted benzyloxy is excluded; and R3 is N-(1-adamantyl)carbamoyl, N-(2-adamantyl)carbamoyl and N-(3-noradamantyl)carbamoyl. Said compound is an 11β-hydroxysteroid dehydrogenase type 1 inhibitor. The invention also relates to a pharmaceutical composition containing said compound as an active ingredient.

EFFECT: improved properties of the compound.

23 cl, 72 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of formula (I): where R1 and R2 represent hydrogen and a group which is hydrolysed in a physiological environment, optionally substituted lower alkanoyl or aroyl; X represents a methylene group; Y represents oxygen atom; n represents the number 0, 1, 2 or 3 and m represents the number 0 or 1; R3 represents a group of pyridine N-oxide according to formula A, B or C which is attached as shown by an unmarked linking: where R4, R5, R6 and R7 independently represent aryl, heterocycle, hydrogen, C1-C6-alkyl, C1-C6-alkylthio, C6-C12-aryloxy or C6-C12-arylthio group, C1-C6-alkylsulphonyl or C6-C12-arylsulphonyl, halogen, C1-C6-haloalkyl, trifluoromethyl, or heteroaryl group; or where two or more residues R4, R5, R6 and R7 taken together represent an aromatic ring, and where P represents a central part, preferentially chosen from regioisomers 1,3,4-oxadiazol-2,5-diyl, 1,2,4-oxadiazol-3,5-diyl, 4-methyl-4H-1,2,4-triazol-3,5-diyl, 1,3,5-triazine-2,4-diyl, 1,2,4-triazine-3,5-diyl, 2H-tetrazol-2,5-diyl, 1,2,3-thiadiazol-4,5-diyl, 1-alkyl-3-(alkoxycarbonyl)-1R-pyrrol-2,5-diyl, where alkyl is presented by methyl, thiazol-2,4-diyl, 1H-pyrazol-1,5-diyl, pyrimidine-2,4-diyl, oxazol-2,4-diyl, carbonyl, 1H-imidazol-1,5-diyl, isoxazol-3,5-diyl, furan-2,4-diyl, benzole-1,3-diyl and (Z)-1-cyanoethene-1,2-diyl, and where the regioisomers of the central part include both regioisomers produced by exchanging the nitrocatechol fragment and the -(X)n-(Y)m-R3 fragment. Also, the invention refers to a method for making a compound of formula I, as well as to a method for treating an individual suffering central and peripheral nervous system disorders, to a pharmaceutical composition based on the compounds of formula I, and also to their application for preparing the drug and as COMT inhibitor.

EFFECT: there are produced and described new compounds which show a potentially effective pharmaceutical properties in treating a number of central and peripheral nervous system disorders.

25 cl, 64 ex, 3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to compounds of formula (I) or to their pharmaceutically acceptable salts, in which X is selected from group, consisting of-C(R1)2-, -O-, -S-, -S(O2)-, -NR1-; each R1 is independently selected from group consisting of H and alkyl; each of R2, R3 and R4 is independently selected from group consisting of (1) H, (2) alkyl, (3) -OR5, (4) alkylene-OR5, (5) -alkylene-R6, (6) -C(O)O-alkyl, (7) - alkylene-C(O)O-alkyl, (8) -alkylene-R8, (9) -NHR5, (10) -N(R5)2, (11) alkenyl, (12) -NH-R8, (13) -NH-CH(C(O)O(C1-C6)alkyl)-alkylene-O-alkyleneR6, (14)-NHCH(C(O)O(C1-C6)aalkyl)-alkylene-OH, (15) -NH-C(O)-alkenyl and (16) -N(C1-C6alkyl)C(O)-alkenyl; or R2 and R3 or R2 and R4 or R3 and R4 together with atoms with which they are bound, form condensed 3-7-member cycloalkyl or heterocycloalkyl ring, which represents non-aromatic monocyclic ring system, which contains in ring from about 5 to about 7 atoms, and one or several atoms in ring system represent atom of element, different from carbon, for instance, nitrogen or oxygen, and said condensed cycloalkyl or heterocycloalkyl ring is not substituted or is substituted with one or several groups L3 ; and on condition that if X represents -O-, and m equals 1, then, at least, one of R2, R3 or R4 is not H; each R5 is independently selected from group consisting of (1) H, (2) (C1-C6)alkyl, (3) hydroxy-substituted alkyl, (4) R6, (5) R7, (6) -C(O)-(C1-C6)alkyl, (7) -C(O)-(C1-C6)halogenalkyl, (8) -C(O)-R6, (9) -C(O)-R7, (10) -C(O)NH-(C1-C6)alkyl, (11) -C(O)N((C1-C6)alkyl)2, in which each alkyl group is selected independently, (12) -S(O)2-(C1-C6)alkyl, (13) -S(O)2-(C1-C6)halogenalkyl, (14) -S(O)2-R6, (15) -S(O)2-R7, (16) -S(O)2-R8, (17) -alkylene-C(O)-(C1-C6)alkyl, (18) -alkylene-C(O)-(C1-C6)halogen-alkyl, (19) -alkylene-C(O)-R6, (20) -alkylene-C(O)-R7, (21) -alkylene-S(O)2-(C1-C6)alkyl, (22) -alkylene-S(O)2-(C1-C6)halogenalkyl, (23) -alkylene-S(O)2-R6, (24) -alkylene-S(O)2-R7, (25) -alkylene-S(O)2-R8, (26) -alkylene-NHC(O)-(C1-C6)alkyl, (27) -alkylene-NHC(O)-(C1-C6)halogenalkyl, (28) alkylene-NHC(O)-R6, (29) -alkylene-NHC(O)-R7, (30) -alkylene-NHS(O)2-(C1-C6)alkyl, (31) -alkylene-NHS(O)2-(C1-C6)halogenalkyl, (32) -alkylene-NHS(O)2-R6, (33) -alkylene-NHS(O)2-R7, (34) -alkylene-N(alkyl)C(O)-(C1-C6)alkyl, (35) -alkylene-N(alkyl)C(O)-(C1-C6)halogenalkyl, (36) -alkylene-N(alkyl)C(O)-R6, (37) -alkylene-N(alkyl)C(O)-R7, (38) -alkylene-N(alkyl)S(O)2-(C1-Ce)alkyl, (39) -alkylene-N(alkyl)S(O)2-(C1-C6)halogen-alkyl, (40)-alkylene-N(alkyl)S(O)2-R6, (41) -alkylene-N(alkyl)S(O)2-R7, (42) -alkylene-C(O)-NH-(C1-C6)alkyl, (43) -alkylene-C(O)-NHR6, (44) -alkylene-C(O)-NHR7, (45) -alkylene-S(O)2NH-(C1-C6)alkyl, (46) -alkylene-S(O)2NH-R6, (47) -alkylene-S(O)2NH-R7 , (48) -alkylene-C(O)-N((C1-C6)alkyl)2, in which each alkyl group is selected independently, (49) -alkylene-C(O)-N(alkyl)-R6, (50) -alkylene-C(O)-N(alkylene)-R7, (51) -alkylene-S(O)2N((C1-C6)alkyl)2, in which each alkyl group is selected independently, (52) -alkylene-S(O)2N(alkyl)-R6, (53) -alkylene-S(O)2N(alkyl)-R7, (54) -alkylene-OH, (55) -alkylene-OC(O)-NH-alkyl, (56) -alkylene-OC(O)NH-R8, (57) -alkylene-CN, (58) -R8, (59) -alkylene-SH, (60) -alkylene-S(O)2-NH-R8, (61) -alkylene-S(O)2-alkylene-R6, (62) substituted with halogen alkylene, (63) -C(O)OR8, (64) -C(O)O(C1-C6)alkyl, (65) -C(O)R8, (66) -C(O)-alkylene-O-(C1-C6)alkyl, (67) -C(O)NH2, (68) -alkylene-O-(C1-C6)alkyl, (69) -alkylene-R8, (70) -S(O)2-halogen(C1-C6)alkyl, (71) hydroxy-substituted halogen(C1-C6)alkyl, (72) -alkylene-NH2, (73) -alkylene-NH-S(O)2-R8, (74) -alkylene-NH-C(O)-R8, (75) -alkylene-NH-C(O)O-(C1-C6)alkyl, (76) -alkylene-O-C(O)-(C1-C6)alkyl, (77) -alkylene-O-S(O)2-(C1-C6)alkyl, (78) -alkylene-R6 , (79) -alkylene-R7, (80) -alkylene-NH-C(O)NH-(C1-C6)alkyl, (81) -alkylene-N(S(O)2 halogen(C1-C6)alkyl)2, and each -S(O)2 halogen(C1-C6)alkyl fragment is selected independently, (82) -alkylene-N((C1-C6)alkyl)S(O)2-R8 , (83) -alkylene-OC(O)-N(alkyl)2, and each alkyl is selected independently, (84) -alkylene-NH-(C1-C6)alkyl, (85) -C(O)-alkylene-C(O)O-(C1-C6)alkyl, (86) -C(O)-C(O)-O-(C1-C6)alkyl, (87) -C(O)-alkylene-R6, (88) -C(O)-NH-R8, (89) -C(O)-NH-R6, (90) -C(O)-NH-alkylene-R6, (91) -C(O)-alkylene-NH-S(O)2-halogen(C1-C6)alkyl, (92) -C(O)-alkylene-NH-C(O)-O-(C1-C6)alkyl, (93) -C(O)-alkylene-NH2, (94) -C(O)-alkylene-NH-S(O)2-R8, (95) -C(O)-alkylene-NH-S(O)2-(C1-C6)alkyl, (96) -C(O)-alkylene-NH-C(O)-(C1-C6)alkyl, (97) -C(O)-alkylene-N(S(O)2(C1-C6)alkyl)2, and each -S(O)2(C1-C6)alkyl fragment is elected independently, (98) -C(O)-alkylene-NH-C(O)-NH-(C1-C6)alkyl, (99) -alkylene-O-R6, (100) -alkylene-R7, (101) -C(O)OH, (102) -alkylene-N(S(O)2(C1-C6)alkyl)2, (103) -alkylene-C(O)-O-(C1-C6)alkyl, (104) halogenalkyl, (105) halogen, (106) -alkylene-C(O)-NH2, (107) =N-O-(C1-C6)alkyl, (108) =N-O-alkylene-R6, (109) =N-O-alkenyl, (110) -N-O-R6, (111) =N-NH-S(O)2-R6, (112) alkenyl, (113) =R8, (114) -O-C(O)-R9, (115) -O-C(O)-(C1-C6)alkyl, (116)-CN, R6 is selected from group consisting of unsubstituted (C6-C14)aryl, (C6-C14)aryl, substituted with one or several groups L1, unsubstituted (C5-C14)heteroaryl and (C5-C14)heteroaryl, which represents aromatic monocyclic or bicyclic system, which contains in ring from about 5 to about 9 atoms, and one or several atoms in ring system represent atom of element, different from carbon, for instance, nitrogen, oxygen or sulphur, one or in combination, substituted with one or several groups L1; R7 is selected from group consisting of unsubstituted heterocycloalkyl and heterocycloalkyl which represents non-aromatic monocyclic system, which contains in ring from about 4 to about 6 atoms, and one or several atoms in ring system represent atom of element, different from carbon, for instance, nitrogen, oxygen substituted with one or several groups L2; R8 is selected from group consisting of unsubstituted cycloalkyl and cycloalkyl substituted with one or several groups L2; A8 is selected from group consisting of (a) unsubstituted aryl, (b) aryl substituted with one or several groups L1; each group L1 is independently selected fron group consisting of halogen, alkyl, -CN, -CF3, -O-(C1-C6)alkyl, -O-(halogen(C1-C6)alkyl), -alkylen-OH (-CH2OH); each group L2 is independently selected from group consisting of (a) -OH, (b) alkyl, (c) alkyl substituted with one or several groups -OH and (d) piperidyl; each group L3 is independently selected from group consisting of -CN, =O, R5 , -OR5 ; =N-R5 and -N(R5)2; n equals 0, 1, 2 or 3; and m equals 0, 1 or 2; and on condition that in composition of substituent -OR5 fragment R5 and oxygen atom, which it is bound with, do not form group -O-O-; and on condition that in composition of substituents -OR5, =N-R5 and -NHR5 R5 are not -CH2OH, -CH2NH2, -CH2NH-alkyl, -CH2NH-aryl or -C(O)OH. Invention also relates to pharmaceutical composition, as well as to application of one or several compounds by one of ii. 1-125.

EFFECT: obtaining novel biologically active compounds possessing properties of γ-secretase inhibitor.

127 cl, 447 ex, 94 tbl

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