Phosphonoacetate esters taxonomic derivatives, intermediate compounds, anticancer pharmaceutical composition, the method of inhibiting tumor growth in mammals

 

(57) Abstract:

Phosphonoacetate esters taxonomic derivatives of the formula I, where R1- hydroxy, -OCH2(OCH2)mOP(O)(OH)2or-OC(O)ORx; R2'is hydrogen; R2- hydroxy, -OCH2(OCH2)mOP(O)(OH)2, -OC(O)ORxor R2'- fluorine, and R2is hydrogen; R3- H, hydroxy, acetoxy; R4, R5- C1-6alkyl, or Z-R6; Z is a direct bond; R6'- aryl or heteroaryl; Rx- C1-6alkyl; p = 0 or 1; R6, R7together form oxoprop, provided that at least one of R1, R2- -OCH2(OCH2)mOP(O)(OH)2m = 0 or 1, or their pharmaceutically acceptable salts possess antitumor activity. 8 C. and 46 C.p. f-crystals, 5 PL.

This application is a partial continuation of simultaneously applying 08/108015 data inventors, filed August 17, 1993, which in turn is a partial continuation of application U.S. N 07/996455, filed December 24, 1992 /now cancelled/. Application USA N 08/108015 included in this description generally by reference.

The present invention relates to antitumor compounds. In particular, the positions and their use as antitumor agents.

Taxol (paclitaxel) is a natural product extracted from the bark of the Pacific yew, Taxus blevifolia. As it turned out, he has an excellent antitumor activity in animal models (in vivo), and recent research has explained his unique character of actions, including abnormal polymerization turbolink and disruption of mitosis. He currently undergoing clinical trials against cancer of the ovary, breast and other types in the United States and France, and preliminary results have confirmed that he is the most promising chemotherapeutic agent. The results of clinical trials of paclitaxel considered in Rowinsky and Donehower, "The Clinical Pharmacology and Use Antimicrotubule Agents in Cancer Chemotherapeutics", Pharmac. Ther., 52 : 35-84, 1991.

Was recently also found a semisynthetic analogue of paclitaxel under the name Taxotere (trade mark), which showed good antitumor activity in animal models. Currently the Taxotere is also undergoing clinical trials in Europe and the United States of America. The following shows the structure of paclitaxel and Taxotere with traditional molecules of paclitaxel numbering system.

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Taxol: R = Ph; R' = aasma organic solubility in water, that requires preparation of dosage forms in the non-aqueous pharmaceutical carriers. One of the commonly used media is Cremophor EL, which itself has on a person's unwanted side effects. Therefore, a number of research groups were obtained water-soluble derivatives of paclitaxel, which are disclosed in the following reference materials:

(a) Haugwitz and others, U.S. patent N 4942184;

(b) the Kingston and others, U.S. patent N 5059699;

(c) Stella and others, U.S. patent N 4960790;

(d) the Application 0558959 A1 for the European patent, published on 8 September 1993;

(e) Vyas ey al. , Bioorganic & Medicinal Chemictry Letters, 1993, 3: 1357-1360; and

(f) Nicolaou et al., Nature, 1993, 364:464-466.

The compounds in accordance with the present invention are phosphonoacetate esters taxonomic derivatives and their pharmaceutically acceptable salts. Solubility of salts in water facilitates the preparation of pharmaceutical compositions.

The present invention relates to taxonomic derivatives having the formula (A)

T -- [OCH2(OCH2)mOP(O)(OH)2]n(A)

where T is taxonomy fragment carrying on the C13 carbon atom substituted 3-amino-2-hydroxypropanoate; n=1, 2, or 3; m=0 or an integer from 1 to 6 inclusive; the train taxonomie derivatives, having the formula (B):

T' -- [OCH2(OCH2)mSCH3]n(IN)

where T' - T, in which no reacts protected hydroxy-group, m and n, such as those defined for formula (A).

Further, in accordance with the present invention offers the intermediates having the formula (C):

T' -- [OCH2(OCH2)mOP(O)(ORy)2]n(WITH)

where T', m and n, such as those defined for formula (A), and Ry- phosphonoamidate group.

In addition, in accordance with the present invention provides compounds of formula (D):

13-OH-txn -- [OCH2(OCH2)mSCH3]n, (D)

where m and n such as those defined above; and txn - taxonomy fragment; or C13 the metal alkoxide.

In accordance with the present invention it is proposed a method of delaying tumor growth in a mammalian host, comprising introducing into an organism mentioned mammal host is effective against tumors of the amount of the compounds of formula (A).

And in accordance with the present invention features a pharmaceutical composition that contains an effective against tumors of the amount of the compounds of formula (A) and pharmaceutically what do the following definition. "Alkyl" means an unbranched or branched saturated carbon chain having from one to six carbon atoms; examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, ISO-butyl, tert-butyl, n-pentyl, isopentyl and n-hexyl. "Alkenyl" means an unbranched or branched carbon chain having at least one double carbon-carbon bond and having from two to six carbon atoms; examples include ethynyl, propenyl, Isopropenyl, butenyl, Isobutanol, pentanol and hexanol. "Quinil" means an unbranched or branched carbon chain having at least one triple carbon-carbon bond and two to six carbon atoms; examples include ethinyl, PROPYNYL, butynyl and hexenyl.

"Aryl" means an aromatic hydrocarbon having from six to ten carbon atoms; examples include phenyl and naphthyl. "Substituted aryl" means aryl substituted with at least one group selected from C1-6alkanoyloxy, hydroxy, halogen, C1-6of alkyl, trifloromethyl, C1-6alkoxy, aryl, C2-6alkenyl, C1-6alkanoyl, nitro, amino, and amido. "Halogen" means fluorine, chlorine, bromine and iodine.

"Phosphono-" means the group-P(O)()mOP(O)(OH)2. "(Methylthio)thiocarbonyl" means the group-C(S)SCH3. "Methylthiomethyl" (also presented as a reduction MTM) is trepiline the name of the group-CH2SCH3.

"Taxonomy fragment" (also present as a reduction txn) refers to fragments containing taxonomy skeleton of twenty carbon atoms represented by the structural formula shown below in absolute configuration.

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Shown above, the numbering system used in this description, is a traditional item taxan, for example, designation C1 refers to the carbon atom marked with the numeral "1"; C5-C20 oxetan refers to oxetanone ring, provided y carbon atoms 4, 5, and 20 oxygen atom; and (C9the hydroxy to the oxygen atom attached to the carbon atom marked with the number "9", and referred to the oxygen atom can be represented by exography, or hydroxy, or - or - acyloxy.

"Substituted 3-amino-2-hydroxypropanoate" means the residue represented by a formula

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(X is a non-hydrogen group and X' is hydrogen or non-hydrogen group). The stereochemistry of this balance is the same as the side chain of paclitaxel. This group can be the I to the connection, having taxonomy fragment bearing a side chain at C13.

"Heteroaryl" means a five - or six-membered aromatic ring containing from at least one to four non-carbon atoms selected from oxygen, sulfur and nitrogen. Examples of heteroaryl include thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, isothiazolin, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, tetrazolyl, oxadiazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, tetrazines and such rings.

"Phosphonate group" means fragments, which can be used to lock (protect) phosphonic functional groups; preferably such protective groups are those that can be removed by methods that do not have a noticeable effect on the rest of the molecule. Suitable phosphonoacetate groups are well known to experts in the art group, including, for example, benzyl and allyl group.

"Hydroxyamine group" include (but are not limited to) ethers, such as methyl, tert-butyl, benzyl, para-methoxybenzyloxy, para-nitrobenzyl, allyl, tritanomaly and trialkylsilyl esters, such as trimethylsilyloxy and tert-butyldimethylsilyloxy ethers; esters, such as benzoyloxy, acetyloxy, phenacyloxy, formally, mono-, di - and trichloracetate, such as chloracetamide, dichloroacetylene, trichloracetate, trifurcation; and carbonates, such as methyl-, ethyl-, 2,2,2-trichloroethyl-, allyl-, benzyl - and paranitrophenylphosphate.

Further examples of hydroxy - and phosphonoacetic groups can be found in standard reference materials, such as Green and wuts, Protective Groups in Organic Synthesis, 2-nd Ed., 1991, John Wiley and Sons, and McOmie, Protective Groups in Organic Chemistry, 1975, Plenum Press.

In these books you can find methods of introducing and removing the protective groups.

"Pharmaceutically acceptable salt" means a metal or amine salt of acid phosphonopropyl, in which the cation does not contribute any significantly to the toxicity or biological activity of the active compounds. Suitable metal salts include lithium, sodium, potassium, calcium, barium, magnesium, zinc and aluminum salts. The preferred metal salts are the sodium and potassium salts. Suitable amine salts are, for example, salts of ammonia, tromethamine (TRIS), triethylamine, procaine, benzydamine, lysine, arginine, ethanolamine, and many others. Preferred aminovymi salts are lysine, arginine and N-methylglucamine salt.

In the description and claims-OCH2(OCH2)mTHE OP(O)(OH)2covers and free acid, and its pharmaceutically acceptable salts, if the meaning is not specifically indicates that refers to the free acid.

In accordance with the present invention offers taxonomie derivatives of the formula (A)

T --- [OCH2(OCH2)mTHE OP(O)(OH)2]n(A)

where T is taxonomy fragment carrying on C13 carbon atom substituted 3-amino-2-hydroxypropanoate; n = 1, 2, or 3; m = 0 or an integer from 1 to 6, inclusive,

or their pharmaceutically acceptable salts.

In one embodiment, taxonomy fragment contains at least the following functional groups: C1 - hydroxy, C2 - benzyloxy, C4 - atomic charges, C5 - C20 oxetan, C9 - hydroxy and C11 - C12 double bond.

In a preferred embodiment, taxonomy fragment produced from the residue having the formula

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where R2e'is hydrogen and R2eis hydrogen, hydroxy, -OC(O)Rxor-OC(O)ORx; or R2eis hydrogen and R2e'- fluorine; R3ex; R6eand R7etogether form oxoprop; Rxsuch as the one defined below.

In another embodiment, the side chain at C13 produced from the residue having the formula

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where R1e- hydrogen or-C(O)Rx, -C(O)ORx; R4and R5- independently C1-6alkyl, C2-6alkenyl, C2-6quinil or-Z-R6;

Z is a direct bond, C1-6alkyl or C2-6alkenyl; R6aryl, substituted aryl, C3-6cycloalkyl or heteroaryl; and Rx- C1-6alkyl, optionally substituted with one to six same or different halogen atoms, C3-6cycloalkyl, C2-6alkenyl or a radical of the formula

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where D is a bond or C1-6alkyl; and Ra, Rband Rcis independently hydrogen, amino, C1-6alkylamino, di-C1-6alkylamino, halogen, C1-6alkyl or C1-6alkoxy; p = 0 or 1.

In a preferred embodiment, R4- C1-6alkyl and p = 1 or R4- -Z-R6and p = 0. More preferably R4(O)p- tert-butoxy, phenyl, isopropoxy, n-propyloxy or n-butoxy.

In another preferred embodiment, R5- C2-6alkenyl or-Z-R6and Z and R6such as 3-6 cycloalkyl.

In another embodiment, the compound of formula (A) can be more specifically represented by formula (I)

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where R1- hydroxy, -OCH2(OCH2)mTHE OP(O)(OH)2, - OC(O)Rxor - OC(O)ORx; R2'is hydrogen and R2is hydrogen, hydroxy, -och2(OCH2)mOP(O)(OH)2or-OC(O)ORx; or R2'- fluorine, and R2is hydrogen; R3is hydrogen, hydroxy, acetoxy, -OCH2(OCH2)mOP(O)(OH)2or-OC(O)ORx; one of R6or R7is hydrogen and the other is hydroxy, C1-6alkanoyloxy or-OCH2(OCH2)mOP(O)(OH)2; or R6and R7together form oxoprop; provided that at least R1, R2, R3, R6or R7- -OCH2(OCH2)mOP(O)(OH)2, R4, R5, Rx, m and p, such as those presented above; or its pharmaceutically acceptable salts.

In the compounds of formula (I) examples Rxinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, chloromethyl, 2,2,2-trichlorethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, ethynyl, 2-propenyl, phenyl, benzyl, bromophenyl, 4-AMINOPHENYL, 4-methylaminophenol, 4-were, 4-methoxyphenyl, etc., Examples R4

In one of the preferred options in accordance with the present invention provides compounds of formula (I) in which R5- C2-6alkenyl or-Z-R6and Z and R6- preferably such as defined above. More preferably R5- phenyl, 3-furyl, 3-thienyl, 2-propenyl, Isobutanol, 2-furyl, 2-thienyl or C3-6cycloalkyl.

In another preferred embodiment, in compounds of formula (I) R4- C1-6alkyl, with p = 1; or R4- -Z-R6and Z and R6such as defined above, with p = 0. More preferably R4(O)p- tert-butoxy, phenyl, isopropoxy, n-propyloxy, n-butoxy.

In another preferred embodiment in accordance with the present invention provides compounds of formula (I) in which R1- -OCH2(OCH2)mTHE OP(O)(OH)2. In a more preferred embodiment, R alkyl. In another preferred embodiment, R3is hydroxy or acetoxy.

In another preferred embodiment in accordance with the present invention features a compound of formula (I), where R2- -OCH2(OCH2)mTHE OP(O)(OH)2; R1is hydroxy or-OC(O)Rx; and R3is hydrogen, hydroxy, acetoxy, -OCH2(OCH2)mTHE OP(O)(OH)2or-OC(O)ORx; and Rxsuch as defined above. In a more preferred embodiment, R1is hydroxy or-OC(O)or SIGxand Rx- preferably C1-6alkyl; and R3is hydroxy or acetoxy.

In another preferred embodiment in accordance with the present invention features a compound of formula (I) in which R3- -OCH2(OCH2)mTHE OP(O)(OH)2; R1is hydroxy or-OC(O)ORx; R2'is hydrogen, and R2is hydrogen, hydroxy or-OC(O)ORx; or R2'- fluorine, and R2is hydrogen; and Rxsuch as defined above. In a more preferred embodiment, R1is hydroxy or-OC(O)ORxand Rx- preferably C1-6alkyl. In another more preferred embodiment, R2is hydroxy.

In another preferred embodiment, m = 0 or 1, when phosphonooxy yimlamai salts of the compounds of formula (A) are alkali metal salts, includes lithium, sodium and potassium salts, and salts of amines, including salts, triethylamine, triethanolamine, ethanolamine, arginine, lysine, and N-methylglucamine. More preferred salts are salts of arginine, lysine and N-methylglucamine.

The most preferred options taxonomic derivatives of the formula (A) include the following compounds: (1) 7-O-phosphonomethylglycine; (2) 2'-O-(ethoxycarbonyl)-7-O-phosphonomethylglycine; (3) 2'-O-phosphonomethylglycine; (4) 2',7-bis-O-(phosphonomethyl)paclitaxel; (5) 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)- 3'-(2-furyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine; (6) 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)- 3'-(2-thienyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine; (7) 10-deacetyl-3'-N-desbenzoyl-3'-N-(tert-butyloxycarbonyl)- 10-O-(phosphonomethyl)paclitaxel; (8) 2'-O-phosphonomethylglycine and their respective pharmaceutically acceptable salts, in particular salts of sodium, potassium, arginine, lysine, N-methylglucamine, ethanolamine, triethylamine, and triethanolamine.

The compounds of formula (A) can be obtained from the original (raw) material - taxonomic derived T - [OH]nwhere T and n is the reactive hydroxy-group, present on taksanova fragment or side chain at C13 to ensure the possibility of formation of simple essential communications (phosphonoacetate ether). It is clear that the reactive hydroxy-group may be directly attached to the main chain propenyloxy at C13 (for example, 2'-hydroxy-group of paclitaxel) or taxonomy the skeleton (for example, 7-hydroxy-group of paclitaxel), or it can be present in the substituents on the side chain at the C13 or Deputy on taksanova the skeleton. To obtain the compounds of formula (A) can be used the sequence of reactions shown in scheme I.

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In scheme I, T' - taxonomie derived, in which no reacts hydroxy-group is already locked (protected); Ry- phosphonoamidate group; n and m are such as defined above. Consequently, respectively protected T', having one or more reactive hydroxy groups are first converted into the corresponding methylthiomethyl ether of the formula (B). When using as an example of paclitaxel T' can be a very effective (for the implementation of the 2', 7-besmedimmibre), 7-O-criticalimportance or 2'-O-ethoxycarbonylmethoxy anhydride or dimethyl sulfide and organic peroxide. These reactions are described more fully below.

MTM ether having one intermediary (intermediate) metalinox-link (i.e., the compounds of formula (B), where m = 1) can be obtained in several ways. In one method, the compound of formula (B), where m = 0, enter into a chemical reaction with N-iodosuccinimide (NIS) and methylthiomethyl to continue the chain on one metalinox-link.

T' -- [OCH2SCH3]n+ nCH3SCH2--OH T' -- [OCH2OCH2SCH3]n.

About the connection methylthiomethyl and getting informed in Syn. Comm., 1986, 16 (13): 1607 - 1610.

In an alternative method T-alkoxide (Ad), obtained by treating the compounds of formula (Aa) base, such as n-utility, diisopropylamide lithium or hexamethyldisilazide lithium enter into interaction with chlorotrimethylsilane ether to obtain the compound of formula (B), where m = 1.

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Connection (Ae) is obtained by introduction of methylthiomethyl (obtained from methylthiomethyl by treatment with base, such as n-utility, diisopropylamide lithium or hexamethyldisilazide lithium) in cooperation with chloromethane. Connection (Ae) can also be obtained by treatment with 1,1'-DIH is a) sodium iodide, and then timelocked sodium. About 1,1'-dichloromethylene broadcast reported in Ind. J. Chem., 1989, 28B, page 454 - 456.

In another method, the compound of formula (Ae) enter into interaction with bis(MTM)ether, CH3SCH2OCH2SCH3and NIS and the result is a compound of formula (B), where m = 1.

T' -- [OH]n+ nCH3SCH2OCH2SCH3---> T'--[OCH2OCH2SCH3]n,

Bis(MTM)ether is obtained by the introduction of 1,1'-dichlorodimethyl ether in the interaction with the sodium iodide, and then thiamethoxam sodium.

The procedure described above using methylthiomethyl and NIS can be applied to any reagent with MTM group, to continue the chain on one metalinox-link at a time. For example, the compound of formula (B), where m = 1, can be entered into interaction with NIS to obtain the compounds of formula (B), in which m = 2. The process can be repeated to obtain the compounds of formula (B), in which m = 3, 4, 5 or 6.

In the second stage, shown in figure 1, methylthiomethyl ether is converted into the corresponding protected phosphonoacetate ether. This is carried out by processing MTM ether NIS and protected phosphate HOP(O)(ORy)2. At the third stage of the th phosphonothioic group is benzyl, which can be removed by catalytic hydrogenolysis; hydroxyamine groups, such as trialkylsilyl can be removed by fluoride ion, trichlorocyanuric may be removed by zinc. Removing the protective groups described in textbooks, such as Green and Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, 1991; and McOmic Protective in Organic Chemistry, Plenum Press, 1973. Both stages are described in more detail later part of the description.

A variant of the sequence of reactions shown in scheme I, is presented in scheme II.

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In scheme II, the compound of formula (Aa) enter into a chemical reaction with compound (Ca) and NIS and get the connection formula (C), which then removes the protection and receive the resulting compound of formula (A). The compounds of formula (Ca), in which m = 0, can be obtained by first processing methylthiomethyl base, such as hexamethyldisilazide sodium, lithium or potassium, to get methylthiomethyl; methoxide is then injected into an interaction with a protected chlorophosphate, such as dibenzylammonium, and receive the requested connection. The compounds of formula (Ca), in which m = 1, can be obtained by treatment of CH3SCH2OCH2Cl datasystem phosphate (salt), for example, dibenzalacetone natlaie Iodate connection with the use of sodium iodide prior to the introduction of the interaction with phosphate. In accordance with another variation of the compounds of formula (Ca), in which m = 1, can be obtained by processing ClCH2OCH2Cl with sodium iodide followed by treatment with thiamethoxam sodium, resulting in a gain of CH3SCH2OCH2SCH3; this compound is then treated with NIS ohms and datasystem phosphate, such as Dienzenhofer, and get the desired product. Any of the above-mentioned reagents with MTM group, may be continued on one metalinox-link at a time by introducing the above-mentioned reagent in cooperation with methylthiomethyl and NIS.

In accordance with another method of obtaining the compounds of formula (A) T-alkoxide (Ad) enter into interaction with iodine-phosphate, as shown in scheme III.

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In scheme III infostate compound obtained by introducing into the interaction ClCH2(OCH2)mCl with datasystem phosphate, which gives ClCH2(OCH2)mOP(O)(ORy)2which is then treated with sodium iodide, resulting in the target product.

Another method, suitable for subgroups of compounds of formula (A), in which at least one of phosphonomethoxy associated with taxonomy fragment, showing the and, P - hidroxizina group, txn - taxonomy fragment. The compounds of formula (D) are takanami with 13 - hydroxy-group and one or more methylthiomethyl esters associated directly or indirectly with taxonomy skeleton; also included C13-alkoxides of the metals of the formula (D). Example of compounds of formula (D) is 7-O-methylthiopyrimidin III:

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The combination taxane (D) azetidinone similar to that shown below in scheme VI, and described the procedure for obtaining compounds of formula (Id) can also be applied to obtain the compounds of formula (Ba) [i.e., the compounds of formula (B), in which at least one MTM group linked directly or indirectly with taxonomy fragment], if instead of the compounds of formula (II) in scheme VI to use the compound of the formula (D). Texan (D) is preferably first converted into C13-metal alkoxide such as sodium alkoxide, potassium or lithium (preferred is a lithium alkoxide). Azetidine serves as a precursor to the side chain at C13. After the reaction combination with taxonom remove hydroxyamino group P and, if required, the free hydroxy-group on the side chain can be turned into MTM ether or produced in the ester or carbonate, as described here.

The General process scheme I to obtain the compounds of formula (A) is presented as an example more specifically, in scheme V, which illustrates obtaining the compounds of formula (I') [i.e., the compounds of formula (I) in which m = 0]. The procedure used in this scheme of synthesis, in General applicable to other taxonomy derived, not specifically covered by formula (I). In addition, the procedure in scheme V may be modified in accordance with information contained in this description of ideas by experts in the field of technology to obtain taxonomic derivatives of the formula (A), in which m = 1 or 2.

It should be understood that in scheme V and all other places of the description, the term "hidroxizina group" may include a carbonate (-OC(O)ORx

In scheme V (see end of description) R1ais hydroxy, protected hydroxy, -OC(O)Rxor-OC(O)ORx; R2'is hydrogen and R2ais hydrogen, hydroxy, protected hydroxy or-OC(O)ORx; or R2'is fluorine and R2ais hydrogen; R3ais hydrogen, hydroxy, protected hydroxy, acetoxy or -- OC(O)ORx; one of R6aor R7ais hydrogen and the other is hydroxy, protected hydroxy, or C1-6alkanoyloxy; R6aand R7atogether form oxoprop; provided that at least one of R1a, R2aor R3a, R6aor R7ais hydroxy, R1bis hydroxy, protected hydroxy, -OCH2SCH3, -OC(O)Rxor-OC(O)ORx; R2'is hydrogen and R2bis hydrogen, hydroxy, protected hydroxy, -OCH2SCH3or-OC(O)Rx; or R2'is fluorine and R2bis hydrogen; R3bis hydrogen, hydroxy, protected hydroxy, acetoxy, -OCH2SCH3or-OC(O)ORx; one of R6bor R7bis hydrogen and the other is hydroxy, protected hydroxy, C1-6alkanoyloxy or-OCH2SCH3; or R6band R7btogether form oxoprop; provided that, one of the radicals R1b2OP(O)(ORy)2, -OC(O)Rxor-OC(O)ORx; R2'is hydrogen, and R2cis hydrogen, hydroxy, protected hydroxy, -OCH2OP(O)(ORy)2or-OC(O)ORx; or R2'is fluorine and R2cis hydrogen; R3cis hydrogen, hydroxy, protected hydroxy, acetoxy, -OCH2OP(O)(ORy)2or-OC(O)ORx; one of R6cor R7cis hydrogen and the other is hydroxy, protected hydroxy, C1-6alkanoyloxy or-OCH2OP(O)(ORy)2; provided that at least one of R1c, R2c, R3c, R6cor R7c- -OCH2OP(O)(ORy)2. R1'- hydroxy, -OCH2OP(O)(OH)2'-OC(O)Rxor-OC(O)ORx; R2"'is hydrogen and R2"is hydrogen, hydroxy, -OCH2OP(O)(OH)2or-OC(O)ORx; or R2"'is fluorine and R2"is hydrogen; R3is hydrogen, hydroxy, acetoxy, -OCH2OP(O)(OH)2or-OC(O)ORx; one of R6'or R7'is hydrogen and the other is hydroxy, C1-6alkanoyloxy or-OCH2P(O)(OH)2; provided that at least one of R1', R2', R3', R6'or R7'- -OCH2OP(O)(OH)2. R4, R5and Rxsuch as defined above, and Ry- phosphonoamidate group.

timedelay ether (-OCH2SCH3). This transformation can be done one of two procedures (Ia - dimethylsulfide method) and (Ib - dimethylsulfoxide method). About dimethylsulfide method of conversion of alcohols in methylthiomethyl esters reported in Medina et al., Tet. Lett., 1988, page 3773 - 3776 (relevant parts of this material are included in this description by reference). Dimethylsulfoxide method is a well-known reaction, commonly called the reaction of Pummerer.

It should be noted that the reactivity of the hydroxy-group is different depending on its location on the source material - taksanova derivative of the formula (Ia). Although a total of 2'- hydroxy-group more reaktsionnosposobnykh in acylation reactions than 7-hydroxy-group, which, in turn, more reaktsionnosposobnykh than 10-hydroxy-group, it has been unexpectedly discovered that 7-hydroxy more easily converted into methylthiomethyl ether than 2'-hydroxy-group, Tertiary hydroxy-group at C-1 is usually the least reaktsionnosposobnykh. The difference in reactivity of the hydroxy-group can be used to control the location and extent of methyldiethylamine.

Thus, in the case of the compounds of formula (Ia), where the th 7-O-methylthiomethyl ether. To obtain the compound of formula (Ib) where R1b- methylthiomethyl without becoming 7-hydroxy-group (if it exists) in methylthiomethyl ether, 7-hydroxy-group block traditional hydroxyamino group, such as triethylsilyl. Similarly, 10-methylthiomethyl ether can be obtained also without turning 7 and/or 2'-hydroxyl groups (if they exist) when they block the same or different hydroxyamine groups. Even if you prefer to methyldiethylamine group is 7-hydroxy, it is still desirable to protect the 2'-hydroxy-group, if the target product is 7-monomethylmercury ether.

In addition, you can adjust the reaction conditions so as to promote the formation of bis - or Tris-methyldiethylamine taxonomic derivatives. For example, in the case of paclitaxel increased the reaction time or the use of a larger excess methylthiopyrimidin reagents may lead to a higher content of 2', 7-bis(methylthiomethyl)ester of paclitaxel in a mixture of products.

During the procedure (Ia) in scheme V, a compound of formula (Ia) is treated with dimethyl sulfide and organic peroxide such as benzoyl peroxide. The reaction is carried out Noi for product education; usually the reaction is carried out in a temperature range from about -40oC to the ambient temperature. Dimethyl sulphide and benzoyl peroxide is used in excess relative to the raw taxonomy derivative (Ia), and dimethyl sulfide is used in excess with respect to the benzoyl peroxide.

The relative amount of used raw materials depend on the degree of methyldiethylamine, which should be achieved. So, when one free hydroxy-group raw tecknologi derivative (Ia) must be turned into methylthiomethyl ether, dimethyl sulfide and benzoyl peroxide can be used in up to 10-fold excess relative to taxonomy derivative (Ia), and an excess of dimethyl sulfide with respect to the benzoyl peroxide is preferably equal to about two-three times. In the case where the raw material (Ia) has 2'- and 7-hydroxy-group, the amount of 2',7-bis(methylthiomethyl)-ether increases with increasing relative amounts of dimethyl sulfide and benzoyl peroxide. When the target product is 2',7-bis(methylthiomethyl)ether, dimethyl sulfide, preferably use about 15 - to 20-fold excess relative to the raw taxonomy proizvoda.

In accordance with another alternative, the compound of formula (Ib) can be obtained by chemical interaction of the compounds of formula (Ia) with dimethylsulfoxide and acetic anhydride (procedure Ib). This procedure is suitable for the conversion of not-2'-hydroxy-group in its methylthiomethyl ether. During the procedure (Ib), the compound of formula (Ia) is dissolved in dimethyl sulfoxide, and the solution was added acetic anhydride. The reaction is usually carried out at room temperature for 18 to 24 hours, and the result is monomethylmercury ether.

In the second stage, the sequence of reactions methylthiomethyl ether is converted into the corresponding protected phosphonoacetate ether. Turning methylthiomethyl ether protected phosphonoacetate ether can be carried out by the General method described in Veeneman et al., Tetrahedron, 1991, v 47, pages 1547 - 1562 (this material is included in this description by reference). In this case, the compound of formula (Ib) with at least one methyldiethylamine group treated with N-iodosuccinimide and protected phosphoric acid, such as Dienzenhofer. The reaction is carried out in an inert organic solvent such as tetrahydrofuran or a halogenated the th agent, such as molecular sieves (zeolites). To accelerate the reaction may also be added to the catalyst, such as triftorbyenzola silver. The reaction is carried out at a temperature in the range of from about 0oC to about room temperature, preferably at room temperature. N-iodosuccinimide and protected phosphoric acid is used in approximately the same molar equivalent of that and methylthiomethyl (Ib), but preferably they are used with a slight excess, for example, approximately 1.3 - 1.5 equivalents, relative to compound of formula (Ib).

In the third stage, the sequence of reactions remove phosphorazidate group and hydroxyamino group (if any). Removing the protective groups is carried out by traditional methods, well known in the art, such as hydrolysis, catalyzed by acid or base, hydrogenolysis, recovery, etc., for Example, catalytic hydrogenolysis can be used for removal of the benzyl phosphonates group, and benzyloxycarbonyl hydroxyamino group . Methods of removing protection can be found in ordinary textbooks, for example, the above-mentioned textbooks green and Votca or Mac-Phi.

Basic salts with which the notes of the compounds of formula (I) with a metal base or amine. Suitable metal bases include the hydroxides, carbonates and bicarbonates of sodium, potassium, lithium, calcium, barium, magnesium, zinc and aluminum, and suitable amines include triethylamine, ammonia, lysine, arginine, N-methylglucamine, ethanolamine, procaine, benzathine, dibenzylamine, tromethamine (TRIS), chloroprocaine, choline, diethanolamine, triethanolamine, etc., Basic salts can then be purified by chromatography was carried out with subsequent lyophilization or by crystallization.

Source derived texana

The processes described above can be applied to any taxonomy derivative of the formula T-[OH]nfor the formation of compounds of formula (A). Many examples of T-[OH]nindicated in the literature and some of them are listed below: (a) paclitaxel; (b) Taxotere (trade mark); (c) 10-deacetyltaxol; (d) taxonomie derivatives disclosed in PCT application 93/06079 (published April 1, 1993), having the formula

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where R1- -OR6, -SR7or-NR8R9; R2is hydrogen, alkyl, alkenyl, quinil, aryl or heteroaryl; R3and R4is independently hydrogen, alkyl, alkenyl, quinil, aryl, heteroaryl or acyl, provided, however, that R3and R4are not both acyl; R5- -COR10the sludge, alkenyl, quinil, aryl, heteroaryl, hidroxizina group or a functional group which increases the solubility in water tecknologi derived; R7- alkyl, alkenyl, quinil, aryl, heteroaryl or sulphadimidine group; R8is hydrogen, alkyl, alkenyl, quinil, aryl, heteroaryl; R9- aminosidine group; R10- alkyl, alkenyl, quinil, aryl, heteroaryl; R11- alkyl, alkenyl, quinil, aryl, heteroaryl, -OR10or-NR8R14; R12and R13- independently from each other alkyl, alkenyl, quinil, aryl, heteroaryl, -OR10or-NR8R14; R14is hydrogen, alkyl, alkenyl, quinil, aryl, heteroaryl; R15and R16- independently from each other hydrogen, hydroxy, lower alkanoyloxy, alkanoyloxy, alkanoyloxy, arrojarse or R15and R16together form oxoprop; R17and R18- independently from each other hydrogen, hydroxy, lower alkanoyloxy, alkanoyloxy, alkanoyloxy, arrojarse or R17and R18together form oxoprop; R19and R20is independently hydrogen or hydroxy or lower alkanoyloxy, alkanoyloxy, alkanoyloxy or arrojarse; R21and R22is independently hydrogen or lower alkanoyloxy, alked or hydroxy or lower alkanoyloxy, alkanoyloxy, alkanoyloxy or arrojarse; or R23and R24together form an oxo or methylene or R23and R24together with the carbon atom to which they are attached, form oxirane ring, or R23and R22together with the carbon atom to which they are attached, form oxetanone ring; R25is hydrogen, hydroxy or lower alkanoyloxy, alkanoyloxy, alkanoyloxy or arrojarse; or R26is hydrogen, hydroxy, or lower alkanoyloxy, alkanoyloxy, alkanoyloxy or arrojarse; or R26and R25together form oxoprop; and R27is hydrogen, hydroxy or lower alkoxy, alkanoyloxy, alkanoyloxy, alkanoyloxy or arrojarse; (e) taxonomie derivatives disclosed in U.S. patent N 5227400: 3'-destiny-3'-(2-furyl)- or 3'-(2-thienyl)derivatives of paclitaxel, Taxotere; (f) taxonomie derivatives disclosed in EP 534709, published March 31, 1993 (derivatives of paclitaxel, in which the phenyl group in the side chain are independently replaced by naphthyl, styryl or substituted phenyl); see also PCT 92/09589, published 11 June 1992; (g) taxonomie derivatives disclosed in EP 534707, published March 31, 1993 (derivatives of paclitaxel, in which 3'-N-benzoline group substituted by paclitaxel and Taxotere); (i) EP 524093, published January 20, 1993 (10-, 7 - or 7,10-bis-O-(N-substituted carbamoylphosphate texana); (j) 9- -hydroxyanisol of paclitaxel disclosed in Klein, "Synthesis of 9-Dihydrotaxol: A New Bioactive Taxane", Tetrahedron Letters, 1993, 34 (13): 20-47-2050; (k) 14- -hydroxyanisol of paclitaxel and Taxotere obtained from 14-hydroxy-10-deacetylbaccatin III, disclosed 205-th national Congress of the American chemical society in Colorado, 1993 (Med. Chem. Division, Abstract N 28); and (l) other taxanes, such as C7-cortexone and various C10-substituted taxanes disclosed in concurrently pending application 08/062687 in U.S. patent submitted by these applicants may 20, 1993, which is fully incorporated into this description by reference.

Free(s) a hydroxy-group(s) taxanomic derivatives can be transformed traditional methods into the corresponding esters or carbonates; for example, in compounds of formula (Ia) one of R1a, R2aor R3a- -OC(O)Rxor-OC(O)ORxand Rxsuch as defined above. Therefore, taxonomie derived T-OH may be entered into interaction with the compound of the formula L-C(O)ORx(L is a leaving group) such as chloroformate, in the presence of a base such as a tertiary amine, to obtain sootvetstvuyuschayatrebovaniyam. T-OH can also be introduced in the interaction with the carboxylic acid RxCO2H or allermuir equivalent (e.g., anhydride, active complex ether or allelochemical) that will give the corresponding ester.

In addition, taxonomie derived T-[OH]ncan be obtained by acylation tecknologi fragment having C13-hydroxy-group, appropriately substituted 3-amino-2-hydroxypropanoic acid, its allermuir equivalent or its predecessor. Suitable precursors of substituted 3-amino-2-hydroxypropanoic acids are, for example, azetidinone formula (III). An example of this acylation reaction is the combination of hydroxyamino baccatin III or hydroxyamino 10-deacetylbaccatin III and derived phenylazomethine that gives derivatives of paclitaxel, as disclosed, for example, issued to Dennis and other U.S. patents N 4924011 and N 4924012; or a combination of protected baccatin III and azetidinone giving paclitaxel and its derivatives, as disclosed in the application 400971 for the European patent, published December 5, 1990 (now U.S. patent N 5175315) and U.S. patent N 5229526.

This process, like the one disclosed in EP 400971 (process Holton) includes m III in the presence of N,N-dimethylaminopyridine and pyridine at 25oC for 12 hours, resulting after removal of the various hydroxyamine groups receive paclitaxel. At Ojima et al. , "New and Efficient Approaches to the Semisynthesis of Taxol and its C-13 Side Chain Analogs by Means of-Lactam Synthon Method," Tetrahedron, 1992, 48 (34): 6985 - 7012 the described improvement of Holton. The process Ojima involves first the formation of the sodium salt of 7-triethylenemelamine III by means of sodium hydride, and then the introduction of this salt in the interaction with chiral 1-benzoyl-3-(1-ethoxy)ethoxy-4-phenyl-2-azetidinone, resulting after removal hydroxyamine groups receive paclitaxel. In U.S. patent N 5229526 Holton discloses the combination of a metal alkoxide baccatin III or its derivative with 2-azetidinone, resulting in receive taxanes with side chain at C13. This process, as mentioned, is highly diastereoselective, therefore, can be used racemic mixture of the precursor 2-azetidinone with the side chain. As recently reported Ojima et al., "A Highly Efficient Route to Taxotere by the-Lactam Synthon Method", Tetrahedron Letters, 1993, 34(26): 4149 - 4152, the combination of metallurgical 7,10-bis-O-(trichlorocyanuric)-10-deacetylbaccatin III with chiral 1-(tert-butoxycarbonyl)-4-phenyl-3-(protected hydroxy)-2-azetidinone network after removing the protection Taxotere. Having Silky.

The process of interaction baccatin with azetidinone used to obtain compounds of formula (Ia) shown in scheme VI (see end of description). This method can also be obtained by using appropriate raw materials other taxonomie derivatives are not specifically covered by the formula (Ia).

In scheme VI, R2'is hydrogen, and R2dis hydrogen, protected hydroxy or-OC(O)ORx; or R2'- fluorine, and R2dis hydrogen; R3dis hydrogen, acetoxy, protected hydroxy or-OC(O)ORx; one of R6dor R7dis hydrogen and the other is hydroxy, protected hydroxy, or C1-6alkanoyloxy; or R6dand R7dtogether form oxoprop; P - hidroxizina group; M is hydrogen or a metal of group 1A, such as lithium, sodium or potassium; and p, R4, R5and Rxsuch as those defined above. The reaction can be carried out in accordance with the procedure disclosed in EP 400971, which is derived baccatin III of the formula (II), where M is hydrogen, enter into interaction with azetidinone formula (III) in the presence of organic bases such as N,N-dimethylaminopyridine. However, it is preferred to first turn derived baccatin III 13-alkoxide by as disclosed above in U.S. patent N 5229526 and reference material author Ogimi. Is preferable, when 13-alkoxide is an alkoxide of lithium. The formation of the lithium salt may be achieved by interaction of the compounds of formula (II), where M is hydrogen, with a strong metal base, such as diisopropylamide lithium, C1-6alkality, bis(trimethylsilyl)amide lithium finality, lithium hydride or a similar basis.

The reaction mix between taxonom formula (II) and azetidinone formula (III) is carried out in an inert organic solvent, such as tetrahydrofuran, at low temperature in the range from approximately 0oC to -78oC. Azetidinone formula (III) may be used as racemic mixtures, combined with metallocene taxane formula (II) in which M is a metal of group IA, and in this case azetidinone reagent is preferably used in amount of at least 2 equivalents relative to taxonomy reagent, and more preferably from about 3 to about 6 equivalents. Can also be used chiral azetidinone, and in this case, it may be sufficient equivalent of azetidinone relatively texana, but preferably azetidine used in a small excess, for example up to 1.5 EC shall ensure the possibility of selective removal of one or more protective groups without significant impact on other groups; for example, in the compound of formula (Id) R2dand PO both can be triethylsilane, and R3dcan be benzyloxycarbonyl; catalytic hydrogenolysis in the presence of palladium on coal removes benzyloxycarbonylamino group without deleting triethylsilyl group. Thus, hydroxyamine group of compounds of formula (Id) can be selectively removed, which will give the compound of formula (Ia).

The compounds of formula (II) are either known from the literature, for example baccatin III, 10-deacetylbaccatin III and hydroxyamine derivatives, or may be obtained from known compounds by conventional methods, for example, the transformation of the hydroxy-group in the carbonate. Additional compounds of formula (II) can be obtained in accordance with the procedures described below in the section "Obtaining raw materials".

The compounds of formula (III) can be obtained from compounds of formula (IIIa) in accordance with the General method described in EP 400971 and Ojima et al. , Tetrahedron 48:6985-7012, 1992.

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In this case, the compound of formula (IIIa) is first treated with a base, takingan-butylethylenediamine,R4(O)pCO-L, where L is a leaving group, and as a result poluchasa, disclosed in EP 400971, through the intermediate connections-3-acetoxy-4-substituted-2-azetidinone (IIIb), or a method disclosed in U.S. patent N 5229526, through the intermediate connections-3-triethylsilyl-4-substituted-2-azetidinone. In the improved method, the compound (IIIb) can be obtained by condensation of acetoacetanilide with bis-Eminem followed by hydrogenolysis or acid cleavage to remove N-kinoway groups; this method is shown in the diagram below, in which R5'- optionally substituted aryl or heteroaryl group such as furyl or thienyl. This method is disclosed in concurrently pending application, U.S. N 08/052434, filed April 23, 1993, which is incorporated in this description by reference.

< / BR>
Product (IIIb) obtained from these reactions cycloaddition, usually a racemic mixture of two cessationof. The racemic mixture can be separated by conventional methods, such as the transformation into the diastereomers, the differential adsorption on a column filled with chiral adsorbents, or enzymatic. For example, a racemic mixture of compounds of the formula (IIIb) may be introduced into contact with the enzyme, which Cagney group of one enantiomer without affecting the other. (See Brieva et al., J. Org. Chem., 1993, 58:1068-1075, and simultaneously consider the application N 092170 on U.S. patent, filed July 14, 1993, application N 552041 for the European patent, published on July 29, 1993). In accordance with another variant of the racemic mixture may be first subjected to base catalyzed hydrolysis to remove the 3-acyl group and education racemic mixture of the corresponding 3-hydroxy - lactam, after which the racemic mixture of 3 - hydroxy - lactate enter into contact with an enzyme capable of catalyzing the acylation of the hydroxy-group to selectively allievate the hydroxy-group of one enantiomer without affecting the other. Or racemic mixture of 3-hydroxy - lactam can be allerban chiral carboxylic acid, and the resulting diastereomers the mixture can then be separated is known in the art methods with subsequent removal of the chiral auxiliary reagent and results in the desired enantiomer.

Materials Ojima et al., J. Org. Chem., 56:1681-1683, 1991; Tet. Lett. , 33: 5737-5740, 1992, Tetrahedron, 48:6985-7012, 1992 described the synthesis of some chiral azetidinone formula (IIIa) and/or of the corresponding N-(p-methoxyphenyl)-high relative, where P is hidroxizina group t is l, 2-phenylethenyl, 2-(2-furyl)ethynyl, 2-methylpropyl, cyclohexylmethyl, isopropyl, phenethyl, 2-cyclohexylethyl or n-propyl. Relevant to the application of these reference materials is included in this description by reference. Other azetidinone within the definition of formula (III), but not specifically disclosed in the above reference materials can be obtained by experts in the field of technology by other methods well-known in this field.

Biological assessment

Compounds in accordance with the present invention are new antitumor substances, the compounds of formula (A) were assessed when testing for cytotoxicity in vitro and in animal models of tumors in vivo.

Data on cytotoxicity in vitro

Compounds in accordance with the present invention showed cytotoxic activity in vitro against cells HCT-116 and HCT-116/VM46 cancerous growths in the colon of man. Cells HCT-116/VM46 are cells that have been previously selected for resistance to teniposide and Express a phenotype resistant to many drugs, including resistance to paclitaxel. Cytotoxicity was assessed in cells HCT-116 colon cancer kissed), as described by D. A. Scudiero et al., "Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines", Cancer Res. , 48: 4827-4833, 1988. Cells were sown at 4000 cells/cell plates of microtitration with 96 cells, and after 24 hours were added medicines and serially diluted. Incubated the cells at 37oC for 72 hours, and during this period he added tetrazolium dye (Templ). The enzyme dehydrogenase in living cells reduces Templ to a form that absorbs light at 450 nm, which can be quantitatively determined spectrophotometrically by. The greater the absorption, the greater the number of living cells. The results were expressed through the IC50/concentration of drug necessary to deter a proliferation of cells (i.e., the absorption at 450 nm) to 50% of the growth of untreated control cells. The values of the IC50to evaluate compounds in this test are given in table 1 at the end of the description.

It was also verified when tested for cytotoxicity of the compound 7-O-methyldiethylamine (Example 1(a)), which showed IC50= 0,003 μm against HCT-116 and 0.025 μm against HCT-116/VM46.

Antitumor activity in vivo

Hybrid mice Balb /c x DBA2F1(CDF1) el for Screening Antitumor Drugs", Cancer Treatment Reports, 65, No. 3-4 pages 299-312, 1981). The test compounds and standard drug (paclitaxel) was administered intravenously to groups of mice, each group received a connection with a different level of dose, resulting in the estimated three to four different doses of the compound. To assess survival of mice monitored daily until their death or approximately 75 days after implantation of the tumor, if they did not die. One group of mice in the experiment were left untreated for control. Tumors were measured once or twice a week, and the size in millimeters used to assess tumor mass in accordance with the published procedure (ibid).

The average survival time of the treated compounds (T) mice compared with an average survival time of parallel, control (C) mice. The ratio of the two values for each treated with the compounds of the group of mice was multiplied by 100 and expressed as a percentage (i.e.% T/C) in table II (see end of description) for representative compounds. In addition, table II also shows the difference between the average time for the groups treated and the average time for the control group when the tumor growth up to 1 g (expressed as the value of T C in days). The larger the value of T-C, the greater the delay ro the 09 SC.

The compound of example 3 (triethanolamine salt) was then evaluated in mouse and human models xenotransplantion tumors (M109, A2780/ DDP ovarian cancer man, are resistant to this drug called cisplatin, and HCT-116 - cancer human colon) compared with paclitaxel as a positive control experiment. Model A2780 /DDP described by Rose and Basler, In vivo, 1989, 3: 249-254. M was perseval (massirovala) subcutaneously every two weeks in Balb /C mice and implanted subcutaneously in CDF1 mice for evaluation of antitumor activity. A2780 /DDP and HCT-116 were grown in deprived thymus mice and reseeding (every two to three weeks), and therapeutic (curative) experiments. The compound of example 3 was injected intravenously in water or orally in water with a few drops of tween 80 and paclitaxel or suspended in water with tween 80, or dissolved in cremophor/ethanol (50%/50%) and diluted with saline. Mode when testing with M (implantation subcutaneously) was performed once a day for 5 consecutive days, starting 4 days after implantation of the tumor. When testing with xenotransplantation of human tumor compounds were injected 5 times - once in the day by day starting from the moment when the tumor was at the stage of 50 - 100 mg by weight.

When you experiment with A2780/c DDP injected compound of example 3 showed the maximum value of the T-C = 29.8 days at 36 mg/kg/injection. (cf., paclitaxel gave max T-C = 26.3 days at 36 mg/kg/injection.). Orally entered the compound of example 3 gave max T-C = 20 dnych/injection. gave 6 the cure of 7 or 6 out of 8 (respectively) treated mice, and oral introducing the compound of example 3 at 160 or 240 mg/kg/with an introd. cured respectively 6 or 7 out of 8 treated mice. Cure means the absence of tumors in 80 day after implantation.

Compounds in accordance with the present invention are phosphonoacetate esters taxonomic derivatives. Form pharmaceutically acceptable salts exhibit improved solubility in water compared with paclitaxel, which allows a comfortable pharmaceutical manufacturing formulations. Without communication with theory, I think, phosphonoacetate esters in accordance with the present invention are drug precursors (previous pharmaceutical raw material) of paclitaxel or its derivative, and postanovochniy fragment is cleaved upon contact with phosphatase in vivo with education then related compounds. As shown above, the compounds in accordance with the present invention are effective agents in inhibiting tumor growth. Therefore, the present invention relates to a method of suppressing tumor in the body of a mammal, which method includes SS="ptx2">

The compounds of formula (A) in accordance with the present invention can be used in the same way as paclitaxel, therefore oncologist - a specialist in the field of cancer treatment, without the necessary experimentation to assign the appropriate treatment course introduction connections in accordance with the present invention. Dosage, method and timetable of the compounds in accordance with the present invention are not particularly limited and may vary with each specific connection. Thus, the connection in accordance with the present invention can be introduced in any suitable way, preferably parenterally; the dose may, for example, be in the range of about 1-100 mg/kg of body weight or about 20 to 500 mg/m2. The compounds of formula (A) can also be administered orally, while the oral dose may be in the range of about 5-500 mg/kg of body weight. The specific dose will vary depending on specifically prepared compositions, method of administration and the specific location of owner and type of treatment to be tumors. When determining the dose take into account many factors affecting drug action, such as age, weight, sex, diet, and physical SOS is notizie, contains effective against tumors of the amount of the compounds of formula (A) in combination with one or more pharmaceutically acceptable carriers, excipients, diluents or excipients. Examples of the manufacture of forms of paclitaxel or its derivatives can be found, for example, in U.S. patents NN 4960790 and 4814470. For example, the compounds in accordance with the present invention can be prepared in the form of tablets, pills, powder mixtures, capsules, injectable, solutions, suppositories, emulsions, dispersions, food premixes, and in other suitable forms. They can also be made in the form of sterile solid compositions, for example, freeze-dried and, if necessary, combined with other pharmaceutically acceptable excipients. Such solid compositions can be restored sterilized water, physiological saline solution or a mixture of water and an organic solvent, such as propylene glycol, ethanol, etc. or any other sterile injectable medium immediately before use for parenteral administration.

Typical pharmaceutically acceptable carriers are, for example, mannitol, urea, dextran who etilzelluloza, poly/vinyl pyrrolidone/, calcium carbonate, etiloleat, isopropylmyristate, benzyl benzoate, sodium carbonate, gelatin, potassium carbonate, silicic acid. The pharmaceutical preparation may also contain non-toxic auxiliary substances such as emulsifying, preserving, wetting means, etc. as, for example, sorbitanoleat, triethanolamine, polyoxyethylenated, glyceryltrinitrate, dioctylsulfosuccinate sodium, etc.

The following experimental procedures all temperatures, when there is no specific guidance presented inoC. Spectral characteristics of nuclear magnetic resonance (NMR) refer to chemical shifts () are expressed in ppm (M. D.), in comparison with tetramethylsilane was (TMS) as ethanol for comparison. Referred to the relative area for various shifts in the spectral data of proton ASR corresponds to the number of hydrogen atoms of a particular functional type in the molecule. The nature of the shifts in the multiplicity is specified as a broad singlet (SHS), broad doublet (sm), broad triplet (PCs), broad Quartet (SCV), singlet (s), multiplet (m), doublet (d), Quartet (q), triplet (t), doublet of doublets (DD), doublet of triplets (l is (deuterated acetone), DMSO-d6(perdeuteromethoxy), D2O (deuterated water), CDCl3(deuterochloroform) and other conventional deuterated solvents. Description of the spectrum of infrared radiation (IR-spectrum) includes only the wave number (cm-1) acquisitions, identification with the values of the functional groups.

Celite is a registered trademark of F. "Jones Manville products Corporation for diatomaceous earth.

Used in this description of the reduction are conventional abbreviations commonly used in the art. Some of them are: MS (mass spectrometry); SVR (mass spectrometry high resolution); Ac (acetyl); Ph (phenyl); v/v (volume/volume, on/o); FAB (fast atom bombardment); NOBA (meta-nitrobenzyloxy alcohol); min (minute/s); h (h/s); NIS (N-iodosuccinimide); BOC (tert-butoxycarbonyl); CBZ (benzyloxycarbonyl); Bn (benzyl); Bz (benzoyl); TES (triethylsilyl); DMSO (dimethylsulfoxide); THF (tetrahydrofuran); HMDS (hexamethyldisilazane).

Obtaining raw (source) materials

Below are examples of several specific raw materials suitable for producing compounds of formula (A).

Obtaining 1. 10-death is impolitical (140 mg, 0,173 mmol) in dry dichloromethane (3.5 ml) was treated at 0oC pyridine (0,028 ml, 0,346 mmol) and trichlorethylphosphate (0,0724 ml, is 0.260 mmol). After 1 hour at this temperature, removed the cold bath, and the mixture was stirred at room temperature overnight. The solvent evaporated, and the residue was chromatographically on silica gel (30-50% ethyl acetate in hexane), receiving specified in the title compound as a foam (92,3 mg, 46%). Further elution gave unreacted starting material (35 mg, 25%) and 2',10-O-bis/2,2,2-trichlorocyanuric)-10-deacetyltaxol with the release of 16%.

(b) 2',7-O-bis(2,2,2-trichlorocyanuric)-10-deacetoxy-11,12 - dehydroacetate-10,12(18)-Dien

Obtained in stage (a) product (92,3 mg, 0,079 mmol) in dry dichloromethane (2 ml) was treated at room temperature 1,1,2-Cryptor-2-chlortrianisene (0,0384 ml, 0,238 mmol). The solution was stirred overnight. The solvent evaporated, and the residue was purified by column chromatography (25% ethyl acetate in hexane), receiving specified in the title compound as a white powder (42,8 mg, 47,3%).

(c) 10-Deacetoxy-11,12-dehydroacetate-10,12(18)-Dien

Obtained in stage (b) the product (39 mg, 0,034 mmol) was dissolved in methane is heated at 40oC for 1 hour, filtered, and the filtrate was evaporated. Chromatography of the residue with a mixture of 60% ethyl acetate/hexane gave specified in the title compound as a foam (22 mg, 81%).

(d) 10-Dezazetilzefotaksima

Obtained in stage (C) the product (22 mg, 0,028 mmol) in ethyl acetate (0.7 ml) was hydrogenosomal at atmospheric pressure in the presence of palladium on charcoal (10%, 14,7 mg, 0.014 mmol Pd). After 5.5 hours at room temperature, filtered (washing with ethyl acetate), evaporation and chromatography (60% ethyl acetate in hexane) gave specified in the header of the product (15,0 mg, 68%) as a white foam.

Getting 2. 7-Deoxy-7 - forpolitical

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(a) 2'-O-Benzyloxycarbonyl-7-deoxy-7 - forpolitical

Diethylaminoethyl TRIFLUORIDE (GIVE, to 18.7 μl, 0,141 mmol) was dissolved in dry dichloromethane (0.5 ml) and the solution was cooled to 0oC. was Added a solution of 2'-O-(benzyloxycarbonyl)paclitaxel (71 mg, 0,072 mmol) in dichloromethane (1 ml) and the resulting solution was kept at 0oC for 30 min and at room temperature for 4 h Then the reaction mixture was added water (0.15 ml) to quickly stop the reaction, and the resulting mixture was concentrated to obtain a residue. The residue was chromatographically on a column of silica gel (with suiteselector-8-desmethyl-7,8-cyclopropylethanol.

(b) 7-Deoxy-7 - forpolitical

Obtained in stage (a) the mixture of products (89 mg) was dissolved in ethyl acetate (3 ml) and the mixture was stirred in an environment of hydrogen under a pressure slightly above one atmosphere in the presence of palladium on charcoal (10% Pd, 29 mg, or 0.027 mmol). After 12 h the solvent was removed, and the residue was purified by silica gel-chromatography (elution with 40% ethyl acetate in hexane), resulting in 67,7 mg specified in the connection header with 8-desmethyl-7,8-cyclopropylethanol.

Then used the method of HPLC (liquid chromatography high pressure) for the separation of 7-deoxy-7-forpolitical and 8-desmethyl-7,8-cyclopropylethanol.

Equipment

Pump: PE series 4

Column: Shandon hypercarb (graphitized carbon), 7 MK, 100 x 4.6 mm, N 59864750 (information on preparative columns can be obtained from Keystone Scientific, Bellefonte, PA)

Injector: PE ISS-100

Detector: HP-104OM

Conditions

Mobile phase: 85:15 methylene chloride:hexane

The division did not disappear at 80:19:1 methylene chloride:hexane:isopropyl alcohol

Flow rate: 2.5 ml/min

Detector: 254 nm

Diluent: the Sample was dissolved in methylene chloride

Get 3. 7-Deoxy-7 - herbacetin III

o
C in a bath of dry ice and acetone and then processed by diethylaminoethyl by TRIFLUORIDE (GIVE, 1.2 ml, 2.5 equiv). The reaction mixture was allowed to mix for 16 h, when it was gradually warmed to ambient temperature. The resulting suspension was filtered and the filtrate (diluted with 30 ml of ethyl acetate) was washed with a saturated aqueous solution of sodium bicarbonate and then brine. The organic fraction was dried (MgSO4) and concentrated, resulting in untreated (raw) product as a white foam. The crude material partially purified by chromatography on a column of silica gel (elution with 10% CH3CN in CH2Cl2) and received 1.45 g of a mixture of 2'-O-(benzyloxycarbonyl)-7-deoxy-7 - forpolitical and 2'-O-(benzyloxycarbonyl)-8-desmethyl-7,8-cyclopropylethanol (a mixture of 82:181H-NMR).

The above-mentioned compound (1.45 g) was dissolved in ethyl acetate (60 ml) and treated with palladium on coal (300 mg). After shaking for 4 h in an environment of hydrogen under pressure, the C short column of silica gel and concentrated. This gave a mixture of the desired product - 7-deoxy-7 - forpolitical and 8-desmethyl-7,8-cyclopropylethanol - in the form of a white foam (1.24 g, yield 99%, mixture of 90:101H-NMR). This mixture was dissolved in dry methylene chloride (30 ml) and treated with tetrabutylammonium (745 mg, 2.9 mmol, 2 equiv.) and allowed to mix for 6 hours Then the reaction mixture was rapidly cooled acetic acid (1 ml), diluted with additional methylene chloride (30 ml) and washed with saturated aqueous sodium bicarbonate. The organic fraction was dried (MgSO4) and concentrated. The crude mixture (substituted taxonomie kernel) was partially purified by chromatography on a column of silica gel) by elution with 10% CH3CN in CH2Cl2and the result was a mixture with the ratio of 90:10 (when the determination method1H-NMR) of 7-deoxy-7 - -herbacetin III and 8-desmethyl-7,8-cyclopropanation III (510 mg, 60%) as a white foam. The resulting foam was led from hot isopropanol, resulting in 7-deoxy-7 - herbacetin III (in the form of small white needles with a yield 410 mg); so pl. 234 - 236oC (decomposition).

Getting 4. 10-Deacetoxy-7-deoxy-7 - forpolitical

< / BR>
(a) 2'-O-Benzyloxycarbonyl-10-deacetoxy (0,0146 ml, is 0.102 mmol) and then diisopropylethylamine (0,0177 ml, is 0.102 mmol). The reaction mixture was stirred at 0oC 45 min and at room temperature for 12 hours Evaporation of the solvent and chromatography on a column of silica gel (elution with 40% ethyl acetate in hexane) gave to 25.5 mg (yield 81%) indicated in the title compound in the form of foam.

(b) 10-Deacetoxy-7-deoxy-7 - forpolitical

Obtained in stage (a) the product of 25.5 mg, 0,028 mmol) in dichloromethane (0.8 ml) was treated with GIVES-ω (0,0071 ml, by 0.055 mmol). After 45 min at 0oC the reaction was allowed to proceed for 5 h at room temperature. Evaporation of the solvent and chromatography gave 2'-O-benzyloxycarbonyl-7-deoxy-7 - forpolitical in the form of a crude foam. This compound was dissolved in ethyl acetate (1 ml) and stirred in a hydrogen environment at a pressure slightly above one atmosphere in the presence of palladium on charcoal (10%, 8,9 mg) for 12 h at room temperature. Was removed by filtering the catalyst, and the product was chromatographically on a column of silica gel, gave 10 mg (yield 40% over two stages) is specified in the header of the product in the form of foam.

Getting 5. 10-Desacetyl-7-deoxy-7 - forpolitical

< / BR>
A solution of 2', 10-O-bis(2,2,2-is handled diethylaminoethyl by TRIFLUORIDE (0,0266 ml, 0,207 mmol). The solution was stirred at 0oC for 30 min and at room temperature for 4 h the Reaction abruptly stopped by adding water (0.05 ml). The reaction mixture was concentrated, and the residue was purified by chromatography on a column of silica gel (elution 30% ethyl acetate in hexane), resulting in 81 mg (yield 68%) of 2',10-O-bis(2,2,2-trichlorocyanuric)-7-deoxy 7 - forpolitical in the form of foam. This compound (63 mg, 0,054 mmol) was dissolved in methanol (0.5 ml) and acetic acid (0.5 ml) and treated with zinc dust (104 mg, of 1.62 mmol) for 90 min at 45oC. the Reaction mixture was filtered, and the filtrate was concentrated. Chromatography on silica gel (elution with 40% hexane to 60% ethyl acetate) mentioned residue gave 38 mg (yield 86%) indicated in the title compound as a white solid.

Getting 6. 7-Detectibility III

< / BR>
(a) 7-O-[Methylthio)thiocarbonyl]baccatin III

Baccatin III (750 mg, 1,278 mmol) was dissolved in dry tetrahydrofuran (20 ml) and the solution was added to one portion of the imidazole (8,7 mg, 0,128 mmol). Was added at room temperature sodium hydride (50% in mineral oil, 77 mg, 1,597 mmol). After cessation of gas evolution (10 min) was added at one time carbon disulphide (4.6 ml). the whole night. Treatment with ethyl acetate and water gave specified in the title compound in the form of unrefined (crude) oil.

An alternative procedure.

Baccatin III (394 mg, 0,672 mmol) was dissolved in tetrahydrofuran (5 ml) and carbon disulfide (1 ml). To this solution was added sodium hydride (of 40.3 mg, 60%, with 1.009 mmol). We also added a catalytic amount of imidazole. The reaction mixture was stirred at room temperature for 1.5 h, then was added methyliodide (122,8 μl, 2,016 mmol). After 40 minutes, the solvent was removed in vacuo, and the residue was chromatographically on silica gel (elution with 20%-50%-60% ethyl acetate in hexano), with a specified title compound (260 mg, yield 57,2%) with 7-epibatidine (98,5 mg, 25%),

(b) 7-O-[(Methylthio)thiocarbonyl]-13-O-triethylcitrate III

The product of stage (a) in the form of crude oil was dissolved in dry dimethylformamide (5 ml) and treated with imidazole (870 mg, 12,78 mmol) and triethylsilane (2.10 ml, 12,78 mmol) at room temperature for 15 hours was Added water, followed by extraction into ethyl acetate. The organic layer was long washed with water and then dried. Pleskrestore on silica gel (elution with 20% ethyl acetate in hexane) gave written>/P>An alternative procedure

Obtained in stage (a) product (193,4 mg, 0,286 mmol) was dissolved in dry dimethylformamide (2,86 ml). To this solution was added imidazole (77.9 mg, to 1.14 mmol), and then triethylsilane (192 μl, to 1.14 mmol). The reaction mixture was stirred overnight at room temperature. After 12 h the reaction mixture was diluted with ethyl acetate (150 ml). The organic layer was washed with water (3 x 10 ml) and brine (1 x 10 ml), dried and concentrated in vacuum. The residue was chromatographically on silica gel (elution with 20% ethyl acetate in hexano) and got the result specified in the header of the product (163 mg, yield 72,0%).

(c) 7-Deoxy-13-O-triethylcitrate III

Obtained in stage (b) product (182 mg, 0,230 mmol) in dry benzene (5 ml) was heated to 80oC in the presence of anti-hydride (0,310 ml, 1,150 mmol) and 2,2'-azobisisobutyronitrile (AIBN, 10 mg). After 3 h the solution was allowed to cool and the solvent evaporated in vacuum. Chromatography on silica gel (elution with 20% ethyl acetate in hexane) of the resulting residue gave specified in the connection header in the form of butter.

(d) 7-Detectibility III

Obtained in stage (C) the product was dissolved in tetrahydrofuran (5 ml) and was treated with tet is assaulte with ethyl acetate and washing with water and brine, followed by chromatography on silica gel (elution with a mixture (1: 1) of ethyl acetate and hexane) has been specified in the title compound as a white glassy solid (63 mg, the output 58% over two stages).

Getting 7. 10-Datasetexception III

< / BR>
(a) 10-Deacetyl-10-O-(Pantothenate)thiocarbonyl-7-O - triethylcitrate III

7-O-Triethylsilyl-10-deacetylbaccatin III (see Green et. al., J. Am. Chem. Soc 110, page 5917, 1988) (319 mg, 0,485 mmol) was dissolved in dry tetrahydrofuran (5 ml), cooled to -40oC and treated with n-butyllithium (1,58 M hexano, 0,384 ml, 0,606 mmol). After 40 minutes at this temperature was added via syringe undiluted pentecostalcharismatic (0,086 ml, 0,536 mmol). The reaction mixture was stirred at -20oC for 90 min, quickly cooled saturated solution of ameriglide and were extracted with ethyl acetate. The ethyl acetate layer was dried and concentrated. The residue was purified by chromatography on silica gel (elution with 40% ethyl acetate in hexane) and got the result mentioned in the title compound as a foam (320 mg, yield 74%).

(b) 10-Deacetoxy-7-O-triethylcitrate III

Obtained in stage (a) product (119 mg, is 0.135 mmol) was dissolved in dry toluene (3 ml) and treated AIBN-Ohm (2 mg). The solution was degirolami dry nitrogen and then added anti-hydride (by 0.055 ml, 0,202 mmol). After this, the solution was heated at 90oC for 1 is as in hexane), receiving specified in the title compound (87 mg, yield 99%) as a colourless foam.

(c) 10-Datasetexception III

Obtained in stage (b) the product (120 mg, 0,186 mmol) was dissolved in acetonitrile (3.5 ml) and the solution was cooled to -10oC. was Added concentrated HCl (36%, to 0.060 ml) and the solution was stirred 30 min. the Mixture was diluted with ethyl acetate (75 ml) and washed with saturated aqueous sodium bicarbonate and brine, then dried and concentrated. The residue was purified by flash chromatography on silica gel (elution with 70% ethyl acetate in hexane) and received a 10-deacetylbaccatin III in the form of foam (75 mg, yield 76%).

Getting 8. 10-Deacetoxy-7-detectibility III

< / BR>
(a) 7-O[(Methylthio)thiocarbonyl]-10-datasetexception III

10-datasetexception III (75 mg, 0,142 mmol) was dissolved in dry tetrahydrofuran (2 ml) and carbon disulfide (0.5 ml). Then was added sodium hydride (60% in mineral oil, 8.5 mg, 0,213 mol) and the mixture was stirred at room temperature for 2 hours was Added itmean (0,026 ml, 0,426 mmol), and allowed the reaction to proceed overnight. Then solvent was removed, and the residue was purified by chromatography on silica gel (elution with 50 - 70% ethyl acetate, the XI-7-detectibility III

Obtained in stage (a) the product (36 mg, 0,058 mmol) in benzene (1 ml) was heated under reflux in the presence of AIBN (2 mg) and anti-hydride (0,079 ml, 0,290 mmol) in argon for 3 hours, the Concentration of the reaction mixture and flash chromatography on silica gel of the residue (elution with 40% ethyl acetate in hexano), followed by the separation by the method of HPLC (liquid chromatography high pressure) from other components Dali specified in the title compound as a foam (16,8 mg, yield 56%).

An alternative procedure

To a solution of 7-O-[(methylthio)thiocarbonyl] -13-O-triethylenemelamine III (product receiving 6, stage (b), 416,3 mg, 0,527 mmol) in dry toluene (10.5 ml) was added a catalytic amount of AIBN, and the resulting solution was degirolami dry N2within 5 minutes was Added to the anti-hydride (708,7 μl, 2,63 mmol) and the reaction mixture was heated at 100oC for 2 h, after which was added another portion of the anti-hydride (425,3 μl, 1,581 mmol). The reaction mixture was heated for 5.5 hours at 100oC, then allowed to cool to room temperature. Chromatography on silica gel (elution with 20% ethyl acetate in hexano) gave 7-deoxy-10-deacetoxy-13-O-(triethylsilyl)baccatin III (320 mg, exit wound (2 ml) at room temperature was added tetrabutylammonium (766 μl, 1 M, 0,766 mmol). The reaction mixture was stirred 1 h at room temperature. Solvent was removed, and the residue was chromatographically on silica gel (elution with 50-70% ethyl acetate in hexano), receiving specified in the header of the target product (115 mg, yield 87.9 per cent).

9. (3R,4S)-1-tert-Butoxycarbonyl-4-phenyl-3-triethylsilyl-2-azetidinone

< / BR>
To a stirred solution of (3R,4S)-4-phenyl-3-triethylsilyl-2-azetidinone (2,200 g, a 7.92 mmol) in dry tetrahydrofuran (25 ml) of N,N-diisopropylethylamine (1.65 ml, 9,510 mmol, 1.2 equiv.) at 0oC in argon. The solution was stirred 5 min, then was added di-tert-BUTYLCARBAMATE (2,080 g 9,510 mmol, 1.2 equiv.) and 4-dimethylaminopyridine (193,6 mg, 1,581 mmol, 0,20 equiv. ). The reaction mixture was stirred at 0oC 60 min and then was diluted with ethyl acetate (25 ml). The resulting solution was washed with brine, 10% NaHCO3, 10% HCl solution, dried (MgSO4) and concentrated, resulting in the crude compound (oil). This compound was then purified by flash chromatography on silica gel (elution with 15% ethyl acetate in hexano) and got mentioned in the title compound as a white solid (2.4 g, yield 83%).

10. ()-qi is the ETP, a magnetic stir bar and addition funnel, was added hydrobenzoin (30,00 g, 100,5 mmol) and ethyl acetate (150 ml). Stirring under a protective layer of argon, the reaction mixture was cooled to 5oC and added to it triethylamine (16,8 ml, 121 mmol). Then added dropwise over 90 min the solution acetoacetanilide (12,4 ml, 116 mmol) in ethyl acetate (300 ml). After 16 hours at this temperature, the reaction mixture was allowed to warm to 20oC (1.5 h) and moved them into a separating funnel. The organic layer was washed sequentially with a saturated aqueous solution of NH4Cl /150 ml, 100 ml, saturated aqueous NaHCO3(120 ml) and brine (120 ml). For identification purposes specified in the header of the connection can be separated at this stage by drying the organic phase over MgSO4, filtration and removal of solvent in vacuo. This gave ()-CIS-3-atomic charges of-1-[(phenyl) (benzylideneamino)methyl]-4-phenylaziridine-2-he with a quantitative yield of crude product in the form of red glass.

(b) the Solution obtained in part (a) compounds in ethyl acetate (500 ml) was carefully transferred under a stream of argon in a 2-liter flask Parra, containing 10% palladium on charcoal (6,00 g). This mixture was treated vodorovne suspended in ethyl acetate (200 ml), stirred (10 min) and filtered. The residue from the filtration was washed with ethyl acetate (100 ml) and the filtrates combined. The organic layer is washed with 10% HCl (300 ml), and both layers were filtered through a funnel with a sintered glass to remove the white precipitate (dibenzylamino HCl), which was washed with ethyl acetate (100 ml). The phases were separated, and the organic layer was washed another part of 10% HCl (200 ml). The combined wash water with 10% HCl was again extracted with ethyl acetate (200 ml) and the combined organic layers washed with saturated aqueous NaHCO3(300 ml) and brine (250 ml). The organic layer was dried over MgSO4, filtered and concentrated in vacuo to a final volume of 75 ml of This mixture was cooled to 4oC, and the precipitated product was separated by filtration. The residue from the filtration was washed with hexane (200 ml), resulting in 16,12 g (78.1% of the total output from hydrobenzoic) specified in the title compound as white needles; so pl. = 150 - 151oC.

Receipt 11. ()-CIS-3-Triethylsilyl-4-(2-furyl)-N-tert - butoxycarbonylamino-2-he

< / BR>
(a) Followed the procedure described in Getting 10, part (a), except that instead of hydrobenzoin, used hydroformed [i.e., 2-furyl-CH-N= CH-2-furyl), the triethylamine (3,11 ml of 22.3 mmol) and acetoxyacetyl (2.30 ml, with 21.4 mmol) gave 6,192 g (yield 90.4 percent) connection ()-CIS-3-atomic charges of-1-[(2-furyl)(2-feniletilamina)-methyl] -4- (2-furyl)azetidin-2-it is in the form of a pale-red syrup.

(b) Followed the procedure described in Getting 10, part (b), except that the product was isolated by preparative thin-layer chromatography, and the reaction was carried out in the calculation of 2.7 mmol, instead of the original number hydroformed. So, obtained in the above part (a) the crude product was again dissolved in ethyl acetate (50 ml) and added to 10% palladium on charcoal (150 mg). Purification of the crude solid by preparative thin-layer chromatography (2 mm silica gel, lirovannomu a mixture of ethyl acetate-hexane to 1:1) gave 386 mg (corrected total yield from hydroformed 65,8%) of ()-CIS-3-(atomic charges)-4-(2-furyl)azetidin-2-it is in the form of a yellow solid. This compound is recrystallized from a mixture of ethyl acetate-hexane; so pl. = 118 - 119oC.

(c) Obtained in the above-described paragraph (b) connection of 3.78 g of 19.4 mmol) in 60 ml of methanol was stirred with K2CO3(20 mg, 0.14 mmol) for 90 min, and the solution was neutralized by Dowex 50w-X8 and f is ivali at 0oC with imidazole (1.44 g, of 21.2 mmol) and TESCl (3.4 ml, at 20.2 mmol) for 30 minutes the Solution was diluted with ethyl acetate and washed with brine, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (elution hexane-ethyl acetate with the ratio 3:1), which gave 4,47 g (yield 86%) of ()-CIS-3-triethylsilyl-4-(2-furyl)azetidin-2-it is in the form of a colorless oil.

(d) Obtained in part (c) product (2,05 g, 7.7 mmol) in 30 ml of dichloromethane was stirred at 0oC diisopropylethylamine (1.5 ml, 8.6 mmol) and di-tert-BUTYLCARBAMATE (2.0 g, 9.2 mmol) and catalytic amount of dimethylaminopyridine (DMAP). The solution was diluted with dichloromethane and washed with brine, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (elution by the mixture hexane-ethyl acetate with the ratio of 8:1) and was obtained 2.0 g (yield 70%) specified in the connection header in the form of a waxy solid.

The racemic mixture obtained in part (b), can be used as a substrate for enzymatic hydrolysis using a lipase, such as PS-30 from Pseudomonas sp. (Amano international Co.), to obtain (3R, 4R)-3-hydroxy-4-(2-furyl)-azetidin-2-it. Method fermentable in USA N 092170, submitted by these applicants on July 14, 1993, which is incorporated by reference in this description completely.

Following the procedures in paragraphs (c) and (d) using (3R,4R)-3-hydroxy-4-(2-furyl)-azetidin-2-it, get the (3R,4R)-N-(tert-butoxycarbonyl)-3-triethylsilyl-4-(2-furyl) azetidin-2-it.

Getting 12. ()-CIS-3-Triethylsilyl-4-(2-thienyl)-N-tert - butoxycarbonylamino-2-he

< / BR>
(a) Followed the procedure described in 10, stage (a), except that instead of hydrobenzoic used hydroceramic [i.e. 2-thienyl-CH-N = CH-2-thienyl)2]. Thus hydroceramic (30 g, 94.7 mmol), triethylamine (15,84 ml, 114 mmol) and acetoxyacetyl (to 11.6 ml, 108 mmol) gave ()-CIS-3-atomic charges of-1-[(2-thienyl) (2 titelmelodie)methyl]-4-(2-thienyl)azetidin-2-it is in the form of a viscous oil.

(b) To mix the solution obtained in part (a) product (0,431 g of 1.03 mmol) in dichloromethane (2,93 ml) at 25oC was added 70% aqueous solution of acetic acid (0.35 ml glacial acetic acid and 0.15 ml of water). The reaction mixture was heated under reflux and stirred for 2.5 hours, the Reaction mixture was diluted with 50 ml dichloromethane and then washed with two 75 ml portions of saturated aqueous solution of bicarbona is of a brown oil, was dissolved in minimum amount of dichloromethane and then was placed on a column of silica gel with dimensions of 4" x 0.5" (10.16 cm x 1.27 cm). Elution with a gradient of 10 to 60% ethyl acetate in hexane gave the less polar by-products and then ()-CIS-3-atomic charges-4-(2-thienyl)-azetidin-2-he (0,154 g, 75% yield) as a white solid.

(c) the solution obtained in part (b) of the product (2.5 g, to 11.8 mmol) was dissolved in methanol (10 ml) and was treated with saturated aqueous sodium bicarbonate (10 ml) and the resulting suspension was allowed to mix at ambient temperature for 3 h Then the reaction mixture was diluted with ethyl acetate (20 ml) and washed with water (15 ml). The aqueous fraction extraction back several times with ethyl acetate, and the combined organic fractions were dried ( MgSO4) and concentrated, resulting in a yellow solid (yield 1.7 g). The crude substance was dissolved in dry tetrahydrofuran (20 ml), and cooled the solution to 5oC in a water bath with ice. After stirring for 5 min was added dropwise triethylchlorosilane (of 1.85 ml, 1.1 equiv.). The resulting suspension was allowed to mix for 3 h at the same temperature, and then separated the ITB is) and concentrated. The crude product was purified by chromatography on a column of silica gel (elution with a mixture of hexane-ethyl acetate to 7:3) and received ()-CIS-3-triethylsilyl-4-(2-thienyl)azetidin-2-it is in the form of a colorless solid (1.5 g, yield 45%); etc. 70 - 71oC.

An alternative procedure

Obtained in part (b) the product (2.0 g, 9,37 mmol) in 40 ml of methanol was stirred with K2CO3(60 mg, 0.43 mmol) for 30 min, and the solution was neutralized by Dowex 50W-X8 and filtered. The filtrate was concentrated, and the residue was dissolved in 50 ml of anhydrous THF and stirred at 0oC with imidazole (0,85 g, 11.3 mmol) and TESCl (1.9 ml, 12.5 mmol) for 30 minutes the Solution was diluted with ethyl acetate and washed with brine, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (elution by the mixture hexane-ethyl acetate with the ratio 3:1), resulting in a 2.13 g (yield 86%) indicated in the title compounds as colorless oils.

(d) the Solution obtained in part (c) product (425,7 mg, 1.48 mmol) was dissolved in dichloromethane (10 ml) and cooled to 5oC in a water bath with ice. The reaction mixture was treated with a catalytic amount of DMAP, and then diisopropylethylamine (TESCl, 0.25 ml, 1.0 in ierature reaction quickly stopped saturated aqueous sodium bicarbonate (45 ml), and the organic fraction washed with water (5 ml), then dried (MgSO4), was passed through a short plug of silica gel and concentrated, with a target product as a colorless oil (525,3 mg, yield 93%).

The procedure described above in Receiving 9, 11 (d) and 12 (d), can be adapted for other N - substituted azetidinone suitable for producing compounds in accordance with the present invention. Examples of such azetidinol are listed in table III; P - hidroxizina group, such as triethylsilyl, triisopropylsilyl and ethoxyethyl.

13. 10-Detoxicated

< / BR>
10-Deacetoxy-7-O-triethylcitrate III (100 mg, 0,156 mmol) was placed in a flask under argon and dissolved in dry tetrahydrofuran (1.5 ml). After cooling to -40oC was added dropwise n-utility (1.45 M in hexano, 0,119 ml, 0,170 mmol), and then (3R, 4S)-1-tert-butoxycarbonyl-4-phenyl-3-triethylsilyl-2-azetidinone cases (94.2 mg, 0.25 mmol) in tetrahydrofuran (0.5 ml) for 2 minutes, the Mixture was immediately warmed to 0oC and stirred for 45 min, before abruptly cooled saturated ammoniacloridegas (3 ml). The mixture was extracted with ethyl acetate, dried and concentrated. Chromatography on Sealy is g, yield 76%). This compound (100 mg, 0,098 mmol) was immediately dissolved in acetonitrile (2 ml) at -5oC and was treated with hydrochloric acid (0.037 ml, 36%, 12 M). The mixture was stirred 2 h at -5oC, and then abruptly cooled water bicarbonate, extracted with ethyl acetate and dried. After evaporation of the solvent the residue was chromatographically on silica gel (elution with 75% ethyl acetate in hexane), giving specified in the title compound as a foam (80,5 mg, yield 80%).

General procedure conducted in Obtaining 13, can be adjusted to obtain other compounds of formula (Ia) with the original components by bakatina III and azetidinone; examples of other compounds of formula (Ia) listed in table IV. It is clear that although the following connections are shown with free hydroxy groups, with appropriate selection of the various hydroxyamine groups any protective groups in position 2', 7 or 10 can be selectively removed without affecting other existing protection group.

Getting 14 IPOs(methylthiomethyl)ether

SH3SCH2OCH2SCH3< / BR>
To a solution of 1,1'-dichloromethylene ester (3.0 g, to 26.3 mmol) in acetone (100 ml) at 0oC was added sodium iodide (8,23 g, 55,23 of molg, 5,23 mmol) and the resulting solution was stirred for another 1 h After this heterogeneous solution was filtered through a layer of celite, and the filtrate was concentrated in vacuum. The residual oil was distributed between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous layer was removed and then was extracted with ethyl acetate, then the combined organics were treated with a mixture of saturated aqueous sodium bicarbonate solution and 5% aqueous sodium thiosulfate solution with a ratio of 1:1 (volume/volume). Then the organics were washed with brine, dried over sodium sulfate and concentrated in vacuum. The residual oil was purified by flash chromatography (30: 1, hexane: ethyl acetate) and got to 1.9 g of a yellow oil which was then distilled using kagalaska apparatus (120 - 130oC, 20 mm RT. Art.) that gave 1.5 g (45%) indicated in the title compounds as a colorless oil:

1H NMR (300 MHz, CDCl3) : to 4.73 (4H, s), of 2.15 (6H, s),

Get 15. Dibenzyldithiocarbamate

CH3SCH2OP(O)(OBu)2< / BR>
To a solution of bis(methylthiomethyl)ester (30 mg, 2.34 mmol) and molecular sieves (300 mg) in tetrahydrofuran (THF) (100 ml) at room temperature was added dimensionful (2,74 g, 9,85 mmol) and then N-IGSCC the Ali through the layer of celite. The filtrate was treated with a mixture of saturated aqueous sodium bicarbonate solution and 5% aqueous sodium thiosulfate solution with a ratio of 1:1 (volume/volume). Then colorless organic extract was washed with brine, dried over sodium sulfate and concentrated in vacuo, resulting in 600 mg (69%) specified in the connection header:

1H NMR (300 MHz, CDCl3) : to 7.35 (10H, s), from 5.29 (2H, d, J = 12,2 Hz) 5,08 (4H, DD, J = 8.0 and 1.0 Hz), and 4.68 (2H, s) of 2.10 (3H, s).

Examples

The following examples are provided to illustrate the synthesis of the presented compounds of in accordance with the present invention and should not be considered as limiting the scope of invention. Specialists in the art can adapt these methods without undue experimentation) to the synthesis of compounds, although not disclosed, but included in the scope of the present invention.

Example 1. 7-O-Phosphonomethylglycine and its monosodium salt

(a) Obtaining 7-O-methyldiethanolamine

< / BR>
To a thoroughly stirred mixture of paclitaxel (0,85 g, 1 mmol) and dimethyl sulfide (to 0.72 ml, 8 mmol) in dry acetonitrile (10 ml) at 0oC was added benzoyl peroxide (0,98 g, 4 mmol). Stirring is continued at Same solvents toluene:ketone (2:1, volume/volume) (Rf.= 0,38, Rf.= 0,64), and when I saw the formation of the products of higher polarity, then the reaction was quickly stopped by evaporation of the solvent using a rotary evaporator Rotavapor at 30oC. Analysis by thin-layer chromatography showed the presence of some amount of unreacted paclitaxel and 2',7-O-bis(methylthiomethyl) of paclitaxel. The Department indicated in the title compound from the reaction mixture produced by column flash chromatography on silica gel 60 (40 - 63 µm). EM Sciense (100 ml), the column diameter of 2 inches (50.6 mm) using the solvent system ethyl acetate-hexane (1: 1, volume/volume) (Rf.=0,34). Product (552 mg, yield 60%) was recovered from fractions 12 - 18 (each faction approximately 20 ml).

Mass spectrum: (FAB/NOBA matrix, NaI, KI): [M+H]+m/z 914; [M+Na]+m/z 936; [M+K]+m/z 952

Elemental analysis: C - 64,28 (calc. 64,39), H - 5,85 (calc. 6,07), N - 1,46 (calc. 1,53).

UV-spectrum (Meon):max= 226 nm (E 1%/1 cm) = 150, A = 0,2653.

IR-spectrum (KBr): 3432, 3066, 2940, 1726, 1668, 1602, 1582, 1514, 1484, 1452, 1372, 1242, 1178, 1142, 1108, 1068, 1026, 990, 916, 884, 852, 802, 774, 710, 608, 570, 538, 482 cm-1.

1H-NMR (CDCl3), : to 1.15 (3H, c), 1,19 (3H, s) of 1.73 (3H, s), 1,79 (H, C) 1,90 (3H, d), is 2.09 (3H, s) of 2.16 (3H, C5,77 (H, DD), 6,16 (H, DD), 6.48 in (H, s), 7,07 (H, d), 7.29 trend is 7.50 (10H, m), EUR 7.57 (H, m), 7,73 (2H, d), 8,08 (2H, d).

(b) Obtain 7-O-dibenzylethylenediamine

< / BR>
To a mixture of 7-O-methyldiethanolamine (119 mg, 0.13 mm) and powdered molecular sieves (about 120 mg) in dry 1,2-dichloromethane (5 ml) was added a solution of N-iodosuccinimide (45 mg, 0.2 mm) and dibenzylamine (55 mg, 0.2 mm) in dry tetrahydrofuran (4 ml). The reaction mixture was stirred at room temperature for 16 hours. The reaction course was monitored by thin-layer chromatography in the system toluene: acetone (2: 1, vol/vol) (Rf.= 0,48). The molecular sieves were removed by filtration through Celite 545 and the filtrate was extracted with methylene chloride (100 ml). The organic layer is washed with 1% sodium thiosulfate solution (about 100 ml) and 0.5 M sodium bicarbonate (100 ml) and brine. The extract was filtered through a separator Paper for separation of the phases, and evaporated solvents. Purification on flash column (silica gel 60 in a mixture of methylene chloride:ethyl acetate (2:1, volume/volume) gave 7-O-dibenzylethylenediamine at 41.5 mg).

(c) Obtain 7-O-phosphonomethylglycine and its monosodium salt

< / BR>
7-O-Dibenzylethylenediamine at 41.5 mg) was dissolved in Atila. inch (275 kPa) at room temperature for 1 hour. The reaction course was monitored by thin-layer chromatography in the system chloroform:methanol:water (120:45:8, vol/vol). Purification by preparative thin-layer chromatography (silica gel carrier plastic HH,05 cm analytical system) gave 7-O-phosphonomethylglycine (26 mg, 75% yield).

Since when cleaning on silica gel watched the decomposition of 7-O - dibenzylethylenediamine, the process of hydrogenation changed. Thus, the crude extract 7-O - dibenzylethylenediamine was hydrogenosomal without any purification. The hydrogenation of the crude extract 7-O-dibenzylethylenediamine was carried out at 60 psi (400 kPa) for 24 hours.

7-O-phosphonomethylglycine (70 mg) was dissolved in 5 ml acetone-water (1:1) and diluted with water to 50 ml) was Added dry sodium bicarbonate (18 mg, 1.2 equiv.). The acetone is evaporated at room temperature using a rotary evaporator and the remaining aqueous solution liofilizirovanny. Untreated monosodium salt of 7-O - phosphonomethylglycine purified by chromatography on a column of C18 reverse phase system water:acetonitrile (70: 30, vol/the reel: of 0.05 M ammonium acetate (45:55, volume/volume), pH=7, Rt=2,09 min Fractions containing the desired product were combined, evaporated acetonitrile, and the remaining aqueous solution liofilizirovanny, resulting in monosodium salt (112 mg), 7-O-phosphonomethylglycine.

Mass spectrum (FAB): [M+H]+m/z 986; [M+Na]+m/z 1008.

UV spectrum (MeOH):max= 230 nm, E (1%/1 cm)=248.

IR-spectrum (KBr): 3430, 3066, 2948, 1724, 1652, 1602, 1580, 1518, 1486, 1452, 1372, 1316, 1246, 1178, 1154, 1108, 1070, 1000, 982, 946, 856, 802, 776, 710, 628, 538 cm-1.

1H-NMR (acetone-d6/D2O): with 8.05 (2H, d), 7,92 (2H, d), the 7.65 (1H, DD), 7,58 - to 7.35 (9H, m, overlapping), of 7.23 (1H, DD), 6,38 (1H, s), between 6.08 (1H, t), the 5.65 (1H, d), the ceiling of 5.60 (1H, d), 5,10 (1H, CL), 4,99 (1H, d), equal to 4.97 (1H, CL), 4,80 (1H, d), to 4.28 (1H, DD), 4,11 (2H, s), with 3.79 (1H, d), to 2.94 (1H, m) to 2.35 (3H, s), 2,35 - 2,10 (1H, m) to 2.13 (3H, s), of 1.95 (3H, s) of 1.84 (1H, m) to 1.67 (3H, s) of 1.13 (6H, s, overlap).

Example 2. An alternative way to obtain 7-O - phosphonomethylglycine

(a) Obtaining 2'-O-(benzyloxycarbonyl)of paclitaxel

< / BR>
To a stirred solution of paclitaxel (150 mg, 0,176 mmol) and N,N-diisopropylethylamine (93 μl, 0,534 mmol, 3 equiv.) in anhydrous methylene chloride (4 ml) at room temperature was added benzylchloride (75 μl, 0,525 mmol, 3 equiv.). The reaction mixture was stirred at room temperature for 3 hours, the oxygen:hexane, as a result, we received the connection specified in the form of a white powder (150 mg, yield 86%); so pl. 140 - 150oC (decomposition).

(b) Obtaining 2'-O-(benzyloxycarbonyl)-7-O - methyldiethanolamine

< / BR>
To a cooled (dry ice-CCl4the bath temperature -30oC) solution of 2'-O-(benzyloxycarbonyl)paclitaxel (4,395 g, 5.0 mmol) in dry acetonitrile (80 ml) was added sequentially dimethyldisulfide (3.6 ml, 40 mmol) and benzoyl peroxide (4.9 g, 20,247 mmol). After 10 minutes at -30oC, cleaned chilled bath and carefully stirred the reaction mixture for 2 h at room temperature. Then the reaction mixture was diluted with ethyl acetate to a volume of 200 ml and washed with water and brine. The organic layer was dried (MgSO4), after which the solvent evaporated, resulting in a residue, which was kept under vacuum for 18 h to remove the DMSO, which was present as a by-product of the reaction. The residue was purified on a column of silica gel using as eluent first mixture (1:2) ethyl acetate-hexane to remove less polar impurities, and then a mixture (1:1) ethyl acetate-hexane, gave the expected result specified in the title compound in the form of foam. Carefully Rastogi substances (5.0 g, 95%); so pl. 120 - 122oC.

Mass spectrum (FAB): [MH]+m/z 1048; [M + Na]+m/z 1070; [M + K]+m/z 108.

IR (KBr): 3440, 3066, 1750, 1722, 1664, 1602, 1583, 1538 cm-1.

NMR (CDCl3), : 1,177 (3H, s), 1,236 (3H, s), 1,745 (3H, s) 2,023 (3H, s), 2,121 (3H, s) 2,162 (3H, s) 2,436 (3H, s) 3,887 (H, d), 4,134 (H, d), 4,197 (H, d), 4,295 (H, m) 4,964 (H, d), 5,161 (2H, d), 5,450 (H, d), 5,703 (H, d), 5,981 (H, DD), 6,257 (H, t) 6,541 (H, C) 6,920 (H, d, NH), 7,322 is 8.22 (15H, m).

Specified in the title compound was also obtained in the following alternative way.

To a solution of 2'-O-(benzyloxycarbonyl)paclitaxel (2.0 g, 2,0263 mmol) in dry dimethylsulfoxide (10 ml) was added dropwise acetic anhydride (10 ml). The resulting mixture was stirred at room temperature for 18 h under N2was diluted with ethyl acetate (100 ml) and thoroughly washed with cold 6% sodium bicarbonate solution (6 x 30 ml), chloride water (6 x 30 ml) and brine. Dried (MgSO4) the organic layer and the solvent is evaporated, after receiving the balance. It was purified on a column of silica gel and was suirable with methylene chloride, a mixture of methylene chloride - 5 % acetonitrile, and a mixture of methylene chloride - 10% acetonitrile, resulting in an expected specified in the title compound (1.86 g, 87.7 per cent). This compound is identical to what has been previously described is bonyl)-7-O-dibenzylethylenediamine

< / BR>
To a solution of 2'-O-(benzyloxycarbonyl)-7-O - methyldiethanolamine (5.0 g, 5,5396 mmol) in dry 1,2-dichloroethane (120 ml) was added activated powdered molecular sieves (5.0 g). To this mixture was added dropwise at room temperature a solution of a mixture of N-iodosuccinimide (1,61 g, 7,1632 mmol) and dibenzylamine (1.97 g, 7,1632 mmol) in dry tetrahydrofuran (90 ml). After intensive stirring at room temperature for 30 min, the reaction mixture was filtered through celite, and the filtrate was evaporated to dryness, receiving a red residue. The residue was dissolved in ethyl acetate (100 ml), washed with cold 6% solution of NaHSO3(2 x 50 ml), cold 6% solution of NaHCO3(2 x 50 ml) and brine (1 x 50 ml). The organic layer was dried (MgSO4), and the solvent evaporated, resulting in a solid mass which was ground into powder with dry diethyl ether and filtered, obtaining mentioned in the title compound in the form of an ivory solid (5.9 g, 97%); so pl. 124 - 127oC.

Mass spectrum (FAB): [MH]+m/z 1278; [M + Na]+m/z 1301; [M + K]+m/z 1316.

IR-spectrum (KBr): 3430, 3066, 3032, 1750, 1726, 1664, 1582, 1532 cm-1.

NMR (CDCl3), : 1,160 (3H, s), 1,703 (3H, s) 1,985 (3H, s) 2,164 (3H, s) 2,420 (3H, s), 3,87,280 - 8,115 (25H, m).

(d) Obtain 7-O-phosphonomethylglycine

To a solution of 2'-O-(benzyloxycarbonyl)-7-O - dibenzylethylenediamine (6.0 g, 4,7095 mmol) in ethyl acetate (120 ml) was added 10% Pd/C (6.0 g) and the mixture was hydrogenosomal at 60 psi (400 kPa) for 24 hours. The reaction mixture was filtered through celite and viparis solvent, got 4,07 g of crude residue. It was purified on a short column of silica gel by successive elution with a mixture of chloroform with 10%, 20% and 40% of methanol and got mentioned in the title compound as a white solid (3.2 g, 71%); so pl. 155 - 158oC.

This product had the same Rf(tondalaya chromatography) and the same retention time (HPLC - liquid chromatography high pressure), and that of the authentic sample.

Mass spectrum (FAB): [MH]+m/z 964; [M + Na]+m/z 986; [M + K]+m/z 1002; [M + K++ Na+- H]+m/z 1024; [M + 2K - H]+m/z 1040.

UV spectrum (MeOH):max= 230 nm, E (1%/1 cm) = 252,5.

IR-spectrum (KBr): 3432, 3066, 2992, 1722, 1648, 1602, 1580, 1522, 1488, 1452, 1372, 1316, 1246, 1178, 1154, 1110, 1070, 1000, 980, 946, 854, 802, 776, 710, 628, 538 cm-1.

1NMR (acetone-d6/D2O): 1,08 (3H, s), 1,10 (3H, s), and 1.63 (3H, s), a 1.88 (3H, s), 1,96 (H, m), 2.13 and (3H), 7,34 - to 7.67 (10H, overlapping multiplets), 7,87 (2H, DD), 8,02 (2H, DD).

Example 3. 2'-O-(Etoxycarbonyl)-7-O-phosphonomethylglycine

(a) Obtaining 2'-O-(etoxycarbonyl)of paclitaxel

< / BR>
To a solution of paclitaxel (4.35 g, 5.1 mmol) in dry methylene chloride (51 ml) was added N,N-diisopropylethylamine or 2.67 ml of 15.3 mmol) and then ethylchloride (1,46 ml of 15.3 mmol). The reaction mixture was stirred at 0oC 2 hours and then at room temperature for another 1 hour. The reaction mixture was diluted with ethyl acetate (400 ml), the organic phase was washed with saturated solution of NaHCO3(2 x 30 ml) and brine (30 ml). The resulting organic phase was dried over MgSO4and received untreated specified in the title compound (93%), which was used in the next stage without additional purification.

Mass spectrum (FAB/NOBA, NaI, KI): [M + H]+m/z 926; [M + Na]+m/z 948; [M + K]+m/z 964.

The mass spectrum of high resolution (FAB/NOBA, external benchmark CSI/Gli): [M + H]+m/z 926,3588 observed, C50H56NO16computed value: 926,3599 (deviation = 1,2 M. D.).

1H-NMR (CDCl3), : of 1.13 (3H, s) of 1.23 (3H, s) of 1.30 (3H, t) to 1.67 (3H, s) of 1.92 (3H, s), of 2.21 (3H, s), 2,37 (H, d), a 2.45 (3H, s) 2,54 (H, m), 3,80 (H, d), 4,15 - 4,32 (4H, overlapping multiplets), 4, multiplet), 7,74 (2H, d), to 8.12 (2H, d).

An alternative procedure

Paclitaxel (of 5.40 g, 6,324 mmol) in dry dichloromethane (63 ml) was cooled to 0oC and was treated with undiluted N,N-diisopropylethylamine (3,30 ml, 3 equiv. and then undiluted etelcharge.com (1,81 ml, 3 equiv.) dropwise within 5 minutes. The reaction was monitored by thin-layer chromatography (50% ethyl acetate in hexane). After 2 h at 0oC and 16 h at room temperature the reaction was completed, and the yellow-orange solution was diluted with ethyl acetate (300 ml) and washed with saturated sodium bicarbonate (3 x 75 ml) and brine (75 ml). Drying (MgSO4) and evaporation gave the crude specified in the title compound, which was purified by precipitation: added dichloromethane (100 ml), then cooled and added hexane (about 60 ml) to the point of turbidity. After cooling in ice for several hours was collected by filtration the solid (5,17 g, yield 88%).

An alternative procedure

In dried over a flame odnogolosy the flask with a capacity of 3 l was dissolved paclitaxel (99,0 g, 115,9 mmol) in 1350 ml of dry methylene chloride in an argon atmosphere. The solution was cooled to -10oC. Slowly (approximately 3 min) was added N,N-diisopropylamino the mixture was stirred overnight (16 hours) at -4oC. Thin layer chromatography showed that the reaction was not completed. So I added another sample of N,N-diisopropylethylamine (2,62 g to 20.28 mmol) and then ClCO2Et (2.20 g, to 20.28 mmol) and continued stirring for 3 hours at -4oC. Thin layer chromatography showed no starting material. The cold mixture was diluted with ethyl acetate (1.5 l) and transferred into a separating funnel. Then it was washed with 5% solution of KHSO4(2 x 500 ml), water (1 x 500 ml), 5% solution of KHSO4(1 x 500 ml), water (1 x 500 ml), a saturated solution of NaHCO3(2 x 500 ml) and brine (2 x 500 ml), dried (MgSO4and by removing the solvent in vacuo, got 147 g of the crude product. The residue was dissolved in hot methylene chloride (800 ml, the temperature of the bath 42oC), was added dropwise hexane (530 ml) with stirring and while maintaining the specified temperature. The crystallizing mixture was set aside for 3 hours at room temperature, and then placed in a cold room (0oC) overnight. Heavy white crystals was collected by filtration and washed with a mixture of hexane - CH2Cl2(1: 1, volume/volume) (2 x 200 ml). After drying in vacuum the filter for 1 hour, they were dried under vacuum (approximately 1.0 mm RT. the measurement by HPLC = 98,5%).

(b) Obtaining 2'-O-(etoxycarbonyl)-7-O-methyldiethanolamine

< / BR>
To a solution of 2'-O-(etoxycarbonyl)paclitaxel (of 4.38 g, 4.7 mmol) in dry dimethylsulfoxide (12.5 ml) was added acetic anhydride (12.5 ml). The reaction mixture was stirred 24 hours at room temperature and then was diluted with ethyl acetate (500 ml), washed with saturated solution of NaHCO3(3 x 40 ml) and water (2 x 40 ml). The obtained organic layer was dried over MgSO4and evaporated the solvent in vacuo to dryness. The residue was purified by chromatography on silica gel (40% ethyl acetate in hexano) and got the desired result is specified in the header connection (4,39 g, 94%).

Mass spectrum (FAB/NOBA, NaI, KI): [M + H]+m/z 986; [M + Na]+m/z 1008; [M + K]+m/z 1024.

MCBP (FAB/NOBA, external benchmark CSI/Gli): [M + H]+m/z 986,3646 (calculated value: 986,3633, reject = 1,3 M. D.).

1H NMR (CDCl3),: : of 1.18 (3H, s) of 1.20 (3H, s) of 1.30 (3H, s) of 1.75 (3H, s), 1,84 (H, m), is 2.09 (3H, s), 2,11 (3H, s) of 2.16 (3H, s), 2,24 (H, d), 2,37 (H, d), a 2.45 (3H, s), 2,80 (H, m), 3,68 (H, d), 4,08 - 4,33 (5H, m, overlapping) and 4.65 (2H, s), 4,96 (H, d), 5,43 (H, d), 5,69 (H, d), 5,98 (H, DD), 6,26 (H, t), 6,55 (H, C) 7,00 (H, d), 7,32 - to 7.61 (11H, m, overlapping), 7,73 (2H, DD), 8,11 (2H, DD).

An alternative procedure

2'-O-(Etoxycarbonyl(paclitaxel (2,260 g, 2,4406 mmol) of p is the anhydride (6 ml). The reaction was monitored by HPLC (analytical C18 column; 60% acetonitrile - 40% 10 mm ammoniumphosphate buffer, pH 6). After 30 h, the solution was diluted with ethyl acetate (250 ml) and washed with saturated aqueous bicarbonate (3 times), and then water and brine. After drying over magnesium sulfate and filtration, the crude product was chromatographically on silica gel (40% ethyl acetate in hexane) and got mentioned in the title compound as a white foam (2,030 g, 91%) with a purity of 90% when determining by HPLC (liquid chromatography high pressure). Part of the product was further purified by a second column (5% acetonitrile in dichloromethane), resulting in the material having a purity of approximately 97% in the determination by HPLC.

An alternative method of obtaining 2'-O-(etoxycarbonyl)-7-O - methyldiethanolamine

2'-O-(Etoxycarbonyl)paclitaxel (4,170 g 4,503 mmol) was dissolved in anhydrous acetonitrile (68 ml) at -40oC, was added dimethyl sulfide (3.2 ml, 44,10 mmol) and then benzoyl peroxide (4,400 g, 18,24 mmol). The mixture was placed in an ice bath and stirred at 0oC, and the reaction course was monitored by thin-layer chromatography (40% ethyl acetate in hexane). After 3 hours, not finding the source material, rest. the ATEM organic phase is washed with bicarbonate, water and brine, then dried over magnesium sulfate and filtered. The residue was purified by flash chromatography on silica gel (4% acetonitrile in dichloromethane) and got the result mentioned in the title compound as a white foam (2,571 g, yield 58%). The purity of this sample was assessed by HPLC and it was over 97%. The NMR spectrum was identical to that presented above.

An alternative procedure for obtaining 2'-O-(etoxycarbonyl)-7-O-methyldiethanolamine

2'-O-(Etoxycarbonyl)paclitaxel (49,3 g, 53.2 mmol) were placed in a dried up over the flame odnogolosy flask with a capacity of 1 liter and was dissolved in dry acetonitrile (500 ml) at room temperature. Through syringe quickly added metilsulfate (39,1 ml, 0,532 mmol). Stirred the reaction mixture was cooled to -16oC in a bath with salt and ice, and to the mixture was added in one portion solid benzoyl peroxide (51,6, 0,213 mmol). (For the full implementation of response required four equivalent). Continued stirring for 30 minutes, during which the temperature rose to about -10oC. In the course of this period, the reaction medium remained heterogeneous (benzoyl peroxide no prior 0oC, and the remaining benzoyl peroxide was dissolved in about 5 minutes after the temperature rise. After stirring for a further 2.5 hours at 0oC reaction, as shown by thin layer chromatography, was completed. By dropping the solution on a rotary evaporator reduces the volume of the solution to approximately 200 ml, and then transferred the solution to a separating funnel where it was washed with heptane (5 x 500 ml). The acetonitrile layer was diluted with ethyl acetate (1.5 l) and washed with a mixture (3:1) saturated NaHCO3and 5% K2CO3(volume/volume) (2 x 500 ml), saturated NaHCO3(2 x 500 ml), polysystem brine (1 x 500 ml) and brine (1 x 500 ml), dried (MgSO4and by removing the solvent in vacuo, got 67,0 g of the crude product. It was dissolved in acetone (200 ml) was heated to 40oC in a water bath, after which was added dropwise with stirring hexane, until not noticed turbidity (400 ml). Crystallizing the mixture was left to stand for 3 hours at room temperature, and then transferred into a cold cell (0oC), where she was kept overnight (16 hours). Formed a thick layer of loose sediment. The solid is collected by filtration and washed with a mixture (3:1, volume/volume) hexane-acetone (2 x 5.5 mm RT. Art.) all night, and the result is received and 47.5 g (yield 91%) specified in the connection header (index homogeneity in the measurement by HPLC was equal to 94.8%).

(c) Obtaining 2'-O-(etoxycarbonyl)-7-O-dibenzylethylenediamine

< / BR>
To a mixture of 2'-O-(etoxycarbonyl)-7-O-methyldiethanolamine (5,677 g, USD 5.76 mmol) and molecular sieves (5.7 g) in methylene chloride (100 ml) at room temperature was added a solution of N-iodosuccinimide (1,953 g, 8,65 mmol) and dibenzylamine (2,41 g, 8,65 mmol) in tetrahydrofuran. The reaction mixture was stirred 40 min at room temperature. After this period the reaction, as shown by thin layer chromatography, was completed. The reaction mixture was filtered through celite, and the filtrate was concentrated in vacuo, resulting in a brownish residue, which was diluted with ethyl acetate (800 ml), the organic phase is washed with 1% Na2SO3(2 x 80 ml) and then with 5% brine (2 x 50 ml). The organic phase was concentrated in vacuum and dried. Chromatography of the resulting residue (50 - 60% ethyl acetate in hexano) gave the target specified in the header connection (6,23 g, 89%).

Mass spectrum (FAB/NOBA, NaI, KI): [M + Na]+m/z 1238; [M + K]+m/z 1254.

The mass spectrum of high resolution (FAB/NOBA,tkanina = 1,3 M. D. ).

1H NMR (CDCl3), : of 1.18 (3H, s) to 1.21 (3H, s) of 1.30 (3H, t) to 1.67 (6H, s), 1,80 (H, s), 1.93 and (H, m), 1,99 (3H, d), to 2.18 (3H, s), 2,23 (H, m), 2,38 (H, m), of 2.45 (3H, s), 2,80 (H, m), 3,86 (H, d), 4,14 - 4,32 (5H, overlapping multiplets), 4,88 (H, d), 5,00 is 5.07 (4H, overlapping multiplets), 5,42 (H, d), 5,68 (H, d), 5,96 (H, DD), 6,26 (H, t) 6,33 (H, s), 6,95 (H, d), 7,30 - to 7.61 (11H, overlapping multiplets), of 7.75 (2H, DD), to 8.12 (2H, DD).

An alternative procedure

To a solution of 2'-O-(Etoxycarbonyl)-7-O-methyldiethanolamine (350 mg, 0,355 mmol) in anhydrous tetrahydrofuran (8 ml) was added a solution of N-iodosuccinimide (120 mg, 0,532 mmol) and dibenzylamine (148 g, 0,532 mmol) in tetrahydrofuran (5 ml). The reaction was monitored by HPLC (C18 column; 70% acetonitrile, 30% 10 mm ammonium phosphate, pH 6). After 2 hours the reaction was complete, as evidenced by less than 5% of the detected source material. The solution was diluted with ethyl acetate (75 ml) and washed with 1% aqueous solution of bisulfite (2 x 50 ml) and brine (50 ml). After rapid drying over magnesium sulfate and filtering, the solvent evaporated. Flash chromatography on silica gel (45% ethyl acetate in hexane) gave specified in the title compound as a white foam (281 mg, 65%). Analysis by HPLC showed a purity of approximately 95%.

An alternative process which is then connected to the vacuum line (approximately 0.5 mm RT.cent.). Sieves was heated by means of a device for drying with hot air for about 10 min with shaking hand. After cooling under vacuum in a flask were introduced argon was added 2'-O-(Etoxycarbonyl)-7-O-methyldiethylamine (37,5 g, 38,03 mmol), and then dimensionful (14.3 g, 53,24 mmol) and tetrahydrofuran (400 ml). The heterogeneous mixture was intensively stirred for 15 minutes at room temperature by a magnetic stirrer. In some dried over a flame flask was dissolved N-iodosuccinimide (10.7 g, 47,54 mmol) in tetrahydrofuran (50 ml) under argon. (During solution preparation NIS transfer fluid and the progress of the reaction vessel was covered with aluminum foil for protection from light). Then to the reaction mixture slowly (10 min) was added via syringe prepared solution. The flask containing the NIS, were washed in 5 ml of THF and transferred in the reaction mixture, which was then stirred 2 hours at room temperature. Analysis by thin-layer chromatography showed no starting material. The solution is dark red was filtered through a layer of celite directly into the intensively stirred biphasic mixture of ethyl acetate (500 ml), 10% aqueous sodium thiosulfate solution (300 ml) and a saturated solution of bi is ivali with ethyl acetate (about 100 ml) and both liquid layer was transferred into a separating funnel. The organic layer was diluted with 1 liter of ethyl acetate, the layers were separated, and the organic layer was washed with a mixture of saturated NaHCO3and 5% K2CO3and then with saturated NaHCO3(2 x 500 ml), polysystem brine (1 x 500 ml) and brine (1 x 500 ml). The extract was dried anhydrous MgSO4and filtered. Then it was treated with 5.0 g of neutral Narita (charcoal) by stirring at room temperature for 15 minutes. Again, it was filtered through a layer of celite, and by removing the solvent under reduced pressure, obtained 52 g of the crude product. It was dissolved in a mixture of toluene and methylene chloride (280 ml/25 ml) and the resulting solution was added dropwise hexane (20 ml). After 3 hours of storage at room temperature for crystallizing the mixture all night kept at 0oC. To the flask walls formed solid pale yellow color. After desantirovaniya mother liquor residue was criterional with toluene (50 ml), filtered, washed with toluene and dried on the vacuum filter for 30 minutes. Then transferred it into a desiccator with drierite, and then dried under vacuum (about 0.5 mm RT.CT.) within 4 hours, resulting in 24.4 g (yield 53%) specified in the header with the, what was retireval with toola (100 ml), filtered, washed with toluene and dried on the vacuum filter for 30 minutes. After drying in a desiccator, as described above, received 12.5 g (yield 27%) of the same product (a measure of homogeneity in the determination by HPLC was equal to 97.1%).

(d) Obtaining 2'-O-(etoxycarbonyl)-7-O-phosphonomethylglycine and its monosodium, montalieu, triethylamine, arginine, lysine, ethanolamine, N-metilglyukaminovoy and triethanolamine salts

< / BR>
To a solution of 2'-O-(etoxycarbonyl)-7-O-dibenzylamino-oxitetraciclina (1,23 g, 1.01 mmol) in dry ethyl acetate (40 ml) was added 10% palladium on coal (428 mg, 10%, 0,404 mmol). The reaction mixture was subjected to hydrogenation at 60 psi (400 kPa) with continuous shaking for 24 hours. The solid was filtered through celite, and then celite washed several times with ethyl acetate. The filtrate was concentrated and the obtained free acid form of the title compound (1.01 g, 80% purity when determining by HPLC). The next phase was removed impurities by preparative chromatography on a column of C-18.

Mass spectrum (FAB/NOBA, NaI, KI): [M + Na]+m/z 1058; [M + K]+m/z ia (FAB/NOBA, external benchmark CSI/Gli): [M + Na]+m/z 1058, 3163

(C51H58NO20PNa, calculated value: 1058, 3188; deviation = 2,3, M. D.).

1H NMR (acetone-d6/D2O), : of 1.13 (3H, s) to 1.21 (3H, s) of 1.66 (3H, s), 1,87 (H, m) of 1.93 (3H, s), and 2.14 (3H, s), 2,18 (H, m) 2,44 (3H, s), 2.95 and (H, m), 3,81 (H, d), of 4.12 (2H, s), 4,15 - 4,27 (3H, overlapping multiplets), 4,92 - 4,99 (2H, overlapping broad multiplets), 5,15 (H, CL), 5,48 (H, d), 5,61 (H, d), of 5.84 (H, DD), 6,07 (H, t) of 6.96 (H, s), 7,25 (H, t), 7,28 - 7,69 (10H, overlapping multiplets), 7,89 (2H, DD), 8,08 (2H, DD), 8,86 (H, d).

An alternative procedure

2'-O-(Etoxycarbonyl)-7-O-(dibenzylideneacetone)- paclitaxel (490 mg, 0,402 mmol) in ethyl acetate (20 ml) was hydrogenosomal in Parr apparatus for shaking at 60 psi (400 kPa) in the presence of palladium on charcoal (10%, weight/mass, 150 mg). The control was carried out by thin-layer chromatography and liquid chromatography high pressure (HPLC). When not found (26 h) no more starting material or intermediate product (preferably monobenzoate), the suspension was filtered through celite and evaporated it to dryness. Analysis by HPLC showed a purity of 88 - 92%.

An alternative procedure

Described below triethylamine salt of 2'-O-(etoxycarbonyl)- 7-O-fo ml) with stirring at 0oC for 30 minutes the Aqueous layer was separated and was extracted with ethyl acetate (20 ml). The combined ethyl acetate layer was washed polysystem brine (25 ml), brine (25 ml x 2), dried over NaSO4and filtered, resulting in an acid solution (approximately of 4.75 mmol) in ethyl acetate (150 ml). This solution is then concentrated to dryness on a rotary evaporator and got 3.75 g specified in the connection header in the form of the free acid form with a 95% yield.

Monosodium salt was prepared as follows.

A sample of 2'-O-(etoxycarbonyl)-7-O-phosphonomethylglycine (1.6 g, 1.55 mmol) was dissolved in acetonitrile (30 ml), subjecting to ultrasound treatment. This solution was diluted with water (30 ml) was added thereto 1.1 M solution of NaHCO3(2,11 ml, 2.32 mmol), alternately shook and were treated by ultrasound, to obtain a solution of (5 - 20 min). The solution slightly milky color was applied on the column, C-18, washed with water in the amount of two volumes of the column, and then suirable monosodium salt 25% acetonitrile in water. The appropriate fractions were combined, evaporated acetonitrile and liofilizirovanny the aqueous phase, resulting in monosodium salt specified in the connection header (850 , m/z 1180.

The mass spectrum of high resolution (FAB / NOBA, external benchmark CSI/Gli): [M + Na]+m/z 1080, 2968 (C51H57NO20PNa2calculated value: 1080, 3007; deviation D=3,6 m D.).

Elemental analysis: C 52,65 (calc. 56,72), H is 5.06 (calc. 5,23), 1,20 N (calc. 1,30), Na 2,74 (calc. 2,12).

IR-spectrum (KBr): 3430, 3066, 2988, 1746, 1722, 1660, 1602, 1582, 1526, 1488, 1452, 1374, 1246, 1178, 1150, 1108, 1070, 1052, 1026, 1002, 966, 912, 834, 792, 776, 710, 628, 538 cm-1.

1H NMR (DMSO-d6D2O-acetone-d6), : 1,10 (6H, c) of 1.23 (3H, t), of 1.64 (3H, c), 1,70 (H, m) 1,90 (3H, s), 1,99 (H, m), and 2.14 (3H, s), is 2.37 (3H, s), 2,98 (H, m), 3,74 (H, d), 4,07 (2H, s), 4,13 - 4.26 deaths (3H, m, overlapping), 4,80 (H, SD), equal to 4.97 (H, d), 5,09 (H, PC), 5,44 (H, d), 5,55 (H, d), 5,99 (H, t) 6,34 (H, s), 7,22 (H, t), 7,43 - 7,69 (10H, m, overlapping), 7,92 (2H, d), of 8.06 (2H, DD).

The sodium salt was prepared as follows.

The crude 2'-O-(etoxycarbonyl)-7-O-phosphonomethylglycine (89%; 70 mg, to 0.060 mmol) in ethyl acetate (2 ml) was treated with a solution of ethylhexanoate sodium (87,5 mm acetate, 1.0 ml, 0,0875 mmol) at room temperature with stirring. After stirring at room temperature for 1 h was added hexane (1.2 ml) to the point of turbidity. After maturation at -20oC for 2 h, was filtered (with some difficulty, very slowly) oncoprotein by HPLC) 95.2% and contained a small amount ethylhexanoic acid (NMR).

Triethanolamine salt was prepared as follows.

The resulting hydrogenation (89% by HPLC 2'-O-(etoxycarbonyl)-7-O-phosphonomethylglycine (0,69 g, 0,593 mmol after correction for impurities) was dissolved in ethyl acetate (10 ml) and slowly stirred while adding dropwise a solution of triethanolamine (0,11 M in ethyl acetate, with the use of 5.1 ml of 0.95 equiv.). Milky white solution obtained by this procedure was degenerately at 0oC for 2 h, then filtered through fine filter paper, washing with cold ethyl acetate. Received 499 mg (80%) of amorphous thin elektrostaticheskogo powder, which was dried overnight in vacuum. HPLC showed 96,6% purity (C-18, 45% of 5 mm Q12+ 10 mm ammonium phosphate pH 6,55% acetonitrile). NMR spectrum (D2O/acetone/DMSO) showed traces of ethyl acetate and the absence of other cuckoobananas impurities. Produced analysis on 2-3 hydrate.

Then was purified triethanolamine salt, previously obtained in the result of another experiment, in the following way. Triethanolamine salt (approximately 2 g) was dissolved in approximately 30% acetonitrile in water. This solution was suirable at low nitrogen pressure through a column of C-18 (Bakerbond) with gradiator by evaporation on a rotary evaporator under reduced pressure. Aqueous solutions were frozen and liofilizirovanny all night, resulting in 1.4 g triethanolamine salt with a purity of 97.5%.

Triethanolamine salt can be obtained as follows.

2'-O-(Etoxycarbonyl)-7-O-phosphonomethylglycine triethylamine salt (3.0 g, of 2.64 mmol) was distributed between ethyl acetate (60 ml) and 5% NaHSO4(30 ml) with intensive stirring at 0oC for 15 minutes. The aqueous layer was separated and was extracted with ethyl acetate (10 ml). The combined ethyl acetate layer was washed with brine (15 ml), dried over Na2SO4, was filtered, and the obtained solution of the acid (about to 2.64 mmol) in ethyl acetate (about 70 ml). To this solution at room temperature was added dropwise with vigorous stirring N(CH2CH2OH)3(0.35 ml, of 2.64 mmol) for 5 minutes. The resulting suspension was stirred for another 1 h, after which it was filtered, washed with ethyl acetate (15 ml × 2), dried under vacuum, and got the 2.8 g triethanolamine salt with a yield of 89%. Analysis by HPLC showed that the index of homogeneity equal to 98.7 per cent; so pl.: above 157oC, with decomposition.

Elemental analysis: calculated for C56H73N2O23P0,3 ethyl acetate: as follows.

To a solution of 2'-O-(etoxycarbonyl)-7-O-dibenzylethylenediamine (10 g, 8,23 mmol) in ethyl acetate (350 ml) at room temperature was added 10% palladium on coal (2 g, 20% loading). The resulting suspension was degirolami by sucking air with the subsequent purge of argon. This process was repeated two more times. Then argon was replaced by hydrogen, and then carried out the same procedure for degassing. The resulting suspension was stirred under hydrogen pressure from a cylinder (2-3 psi, i.e., 0.14 to 0.21 kg/cm2) for 16 h at room temperature with intensive stirring. Was aspirated hydrogen and replaced it with argon three times, followed by the implementation of procedures degassing. The resulting suspension was filtered through a layer of celite. To this homogeneous filtrate was slowly added Et3N (8,23 mmol, 1,14 ml) for 5 minutes under vigorous stirring. The obtained fine white suspension was stirred for another 30 minutes. Was filtered through a funnel with sintered porous filter with a porosity E. the Residue from the filtration was dried under vacuum (1 mm RT.CT.) for 16 h, and was obtained by 8.22 g specified in the header triethylamine salt with a yield of 88%. Analysis by HLC gave pok>7H73N2O20P 4,5 H2O: C 56,19; H 6,79; N 2,30; KF (H2O) of 6.65.

Calculated: C 56,33; H 6,87; N 2,32; KF (H2O) of 7.96.

An alternative procedure for obtaining triethylamine salt.

In a flask with a capacity of 250 ml was added 2'-O-(etoxycarbonyl)-7-O-dibenzylethylenediamine (5,67 g of 4.66 mmol) and dissolving it in ethyl acetate (150 ml). The flask was supplied with three-way valve, one of the connections of which are connected to the vacuum line and the other line of the argon. By using a valve in the flask was creating a partial vacuum and then purged with argon. This process was repeated two more times. To the flask was added palladium on charcoal (10% Pd) (0.85 grams). Line argon attached to the three-way valve was replaced with a balloon filled with hydrogen. Through the valve in the flask had created a vacuum, and then blew it with hydrogen. This process was repeated four more times. The resulting mixture was stirred at room temperature in the environment of hydrogen from the container throughout the night. Analysis by thin-layer chromatography after 17 hours after the initial exposure to hydrogen showed the absence of starting material. The balloon with hydrogen attached to a three-way valve, a shelter. This process was repeated two more times. The contents of the flask were subjected to vacuum filtration through a layer of celite. Celite was washed with ethyl acetate (2 x 10 ml). To the stirred filtrate was added triethylamine (0,650 ml of 4.66 mmol). The resulting suspension was stirred at room temperature for two hours, then was reduced to about 150 ml by evaporation on a rotary evaporator. The solid was filtered, washed with ethyl acetate (2 x 10 ml) and dried under vacuum, resulting in value of 4.76 g (yield 90%) indicated in the title triethylamine salt as a white powder (analysis by HPLC showed that the index of homogeneity of 96.6%).

An alternative procedure for obtaining triethylamine salt.

In a flask with a capacity of 250 ml was injected 2'-O-(etoxycarbonyl)-7-O-dibenzylethylenediamine (of 5.17 g of 4.25 mmol) and dissolving it in ethyl acetate (150 ml). The flask was supplied with three-way valve, one of the connections of which are connected to the vacuum line and the other line of the argon. Using the valve, was created in the flask partial vacuum, and then was purged with argon. This process was repeated two more times. Was added into the flask palladium on charcoal (10% Pd, 0,86 g). Line argon attached to t the th rarefaction, then was purged with hydrogen. This process was repeated five more times. The resulting mixture was stirred at room temperature in the environment of hydrogen from the container throughout the night. Analysis by thin-layer chromatography after 16 hours after the initial exposure to hydrogen showed the absence of starting material. The balloon with hydrogen attached to a three-way valve, replaced by a line of argon. Through valve was created in the flask partial vacuum, and then was purged with argon. This process was repeated two more times. The contents of the flask were subjected to vacuum filtration through a layer of celite. Celite was washed with ethyl acetate (4 x 10 ml). To the stirred filtrate was added triethylamine (0,590 ml of 4.25 mmol). The resulting suspension was stirred at room temperature for one hour, then was reduced to about 140 ml by evaporation on a rotary evaporator. The solid was filtered, washed with ethyl acetate (10 ml) and dried under vacuum, resulting in 4,46 g (yield 92%) indicated in the title triethylamine salt as a white powder (determined by HPLC analysis of the index of homogeneity was 96.7%).

The lysine salt was prepared as follows.

To suspe oxycarbonyl)-7-O-dibenzylethylenediamine (15.0 g, 12,34 mmol). The resulting suspension was degirolami by sucking air and purging with argon. This process was repeated two more times. Then argon was replaced by hydrogen, and then performed the same procedure degassing under intensive stirring at 0oC for 2 hours Remove the cooling bath and the reaction mixture was stirred at ambient temperature for another 4.5 hours. The reaction mixture was degirolami by sucking hydrogen and purge with argon three times, it was filtered under argon through a layer of celite. To the obtained filtrate was slowly added a solution of lysine (1.63 g, 0,94 equiv.) in a mixture of water:ethanol (1:1, the fortress of 200, 20 ml) for 5 minutes under vigorous stirring. To the resulting white suspension was added distilled water (110 ml) and stirred suspension of 30 minutes. Would heat it up to about 55oC. the Obtained homogeneous solution was kept in the installation of the oil bath at 50oC and slowly cooled to room temperature, 16 hours and up to 4oC for 3 hours, filtered it, and subjecting to vacuum drying for 16 hours, got to 11.8 g (yield about 80%) lysine salt having uniformity 99,0% in the determination by HPLC; so pl. above 170oC with the N 3,17; KF (H2O) 10,87.

Found: C 51,76; H to 6.57; N 3,48; KF (H2O) 11,42.

Ethanolamine salt was prepared as follows

2'-O-(Etoxycarbonyl)-7-O-phosphonomethylglycine triethylamine salt (3.0 g, of 2.64 mmol) was distributed between ethyl acetate (60 ml) and 5% NaHSO4(30 ml) with intensive stirring at 0oC for 15 minutes. The aqueous layer was separated and was extracted with ethyl acetate (15 ml). The combined ethyl acetate layer was washed with brine (15 ml), dried over Na2SO4, was filtered, and the obtained solution of the free acid (about to 2.64 mmol) in ethyl acetate (about 70 ml). To this solution at room temperature was added dropwise with intensive stirring, a solution of H2NCH2CH2OH (0.15 ml, of 2.64 mmol) in ethyl acetate (5 ml) for 5 minutes. The resulting suspension was stirred for another 1 hour, after which it was filtered, washed with ethyl acetate (15 ml × 2) and dried under vacuum, resulting in a 2.6 g specified in the header ethanolamine salt with a yield of 89%. Uniformity in the determination by HPLC-analysis was equal to 97.8 per cent; so pl. above 130oC with decomposition.

Elemental analysis: calculated for C53H65N2O21P 2,5 H2Yu Sol was prepared as follows.

To a suspension of 10% palladium on coal (load of 20%, 6 g) in ethanol (900 ml, fortress 200) at 0oC) was added in parts 2'-O-(etoxycarbonyl)-7-O-dibenzylethylenediamine (30.0 g, 24,69 mmol). The resulting suspension was degirolami by sucking air and purging with argon. This process was repeated two more times. Then argon was replaced by hydrogen, after which he again performed the procedure described above degassing under vigorous stirring. The resulting mixture was stirred at 0oC for 2 hours. Removed the cooling bath and the reaction was stirred solution at ambient temperature for another 24 hours. The reaction mixture was degirolami by sucking hydrogen and purge with argon, through the above procedure three times. Was filtered under argon through a layer of celite. The filtrate was divided into two equal parts, and each part was added ethanol (190 ml, fortress 200). To one part (about 630 ml) was slowly added a solution of arginine (2.0 g, 0,94 equiv. ) in a mixture of water: ethanol (2:1, the fortress of 200, 20 ml) for 5 minutes under vigorous stirring. To the resulting white suspension was added distilled water (100 ml) and stirred 30 minutes, then was heated to approximately 60oC. it was Filtered hot, the URS, and kept at room temperature for 2 hours and at 4oC for 2 hours. Filtered it, and washed with cold mixture of 3% water in ethanol (100 ml) and dried by suction for 16 hours, resulting in a received 12,95 g (approximately 86% yield) specified in the header of the arginine salt uniformity equal to 96,7%.

This material (12,95 g) was dissolved in a mixture of 15% water in ethanol (700 ml) at 55oC. the Solution was cooled and kept at 30oC for 3.5 hours, at room temperature for 16 hours and at 4oC for 3 hours. The obtained crystals were filtered off, washed with cold mixture of 2% water in ethanol (50 ml x 2), was subjected to drying by suction for 4 hours and then dried under vacuum (1 mm RT.CT.) 16 hours, resulting in a 10.2 g (80% yield) specified in the header of arginine salt (uniformity of 98.5%); so pl.: above 176oC with decomposition.

Elemental analysis: calculated for C57H72N5O22P 6,4 H2O:

C 51,65; H 6,45; N 5,28; KF (H2O) 8,7.

Found: C 51,86; H of 6.65; N of 5.53; KF (H2O) 8,72.

N-Methylglucamine salt was prepared as follows.

To a suspension of 10% palladium on coal (20% load, 6 g) in metekel (30.0 g, 24,69 mmol). The resulting suspension was degirolami by sucking air and purging with argon. This process was repeated two more times. Then argon was replaced by hydrogen, and then carried out the above procedure with vigorous stirring. The resulting mixture was stirred at 0oC for 2 hours. Removed the cooling bath, and the reaction solution was stirred at ambient temperature for another 24 hours. The reaction mixture was degirolami by sucking hydrogen and purge with argon, following the above procedure three times. Was filtered under argon through a layer of celite. The filtrate was divided into two equal parts, and each part was added ethanol (190 ml, fortress 200). To one part (about 600 ml) was slowly added a solution of N-methylglucamine (2.24 g, 0,94 equiv.) in a mixture of water:ethanol (1: 1, the fortress of 200, 20 ml) for 5 minutes under vigorous stirring. To the resulting white suspension was added distilled water (100 ml), and the suspension was stirred for 30 minutes and then was heated to about 49oC. a Transparent homogeneous solution was kept in the installation of the oil bath at 50oC, allowing it to cool to room temperature, and kept at room temperature for 2 hours and at 4oC for 1.5 hours. Solution filtralite 9,65 g (approximately 64% yield) specified in the header of the N-metilglyukaminovoy salt uniformity 96,4%.

This material (9,65 g) was dissolved in a mixture of 15% water in ethanol (450 ml) at 52oC. Then the solution was cooled and kept at 28oC 3.5 hours at room temperature 16 hours at 4oC 3 hours. The obtained crystals were filtered off, washed with cold mixture of 2% water in ethanol (50 ml x 2), dried by suction for 4 hours and then dried under vacuum (1 mm RT.cent.), that resulted in 7.5 g (about 80% yield) specified in the header of the N-metilglyukaminovoy salt, a measure of uniformity in the determination by HPLC amounted to 98.6%); so pl.: above 154oC with decomposition.

Elemental analysis: calculated for C58H75N2O25R 5,0 H2O:

C 52,72; H 6,48; N 2,12; KF (H2O) 6,82.

Found: C 53,09; H 6,50; N 2,08; KF (H2O) 7,12.

Example 4. 2'-O-(phosphonomethyl)paclitaxel

(a) Obtaining 2'-O-(methylthiomethyl)-7-O-(triethylsilyl)-paclitaxel

< / BR>
To a chilled (0 - (-5oC) a solution of 7-O-(triethylsilyl)paclitaxel (2,46 g, 2,5439 mmol) in dry acetonitrile (100 ml) was added dimethyl sulfide (1,348 g of 1.59 ml, 21,6976 mmol) and then benzoyl peroxide (2,628 g, 10,8488 mmol). The heterogeneous mixture was stirred at 0oC 1 hour and kept at the 5oC 18 hours. Got a yellow solution. It was evaporated the PE the result is listed in the title compound (1.0 g, 38%). It was used as such for the next stage.

Mass spectrum: [M + H]+, 1028; [M + Na]+, 1050; [M + K]+, 1066.

(b) Obtaining 2'-O-(methylthiomethyl)of paclitaxel

< / BR>
To a cooled (-15oC) the solution obtained in stage (a) product (1.0 g, 0,9737 mmol) in dry acetonitrile (30 ml) was added floor drops of 0.5 N. HCl (3 ml). The resulting solution was stirred at -15oC for 1 hour and 5oC for 18 hours. Diluted with ethyl acetate (20 ml) and washed with cold 6% solution NaHCP3and brine. Dried (gSO4) and was evaporated to dryness. The obtained product was purified on coated silica gel plate (methylene chloride to 15% acetonitrile) and received net specified in the title compound (280 mg, 31.4 percent).

IR-spectrum: 3446, 3064, 2940, 1726, 1666, 1582, 1516, 1486 cm-1.

NMR (CDCl3): 1,118 (s, 3H), 1,229 (s, 3H), 1,622 (s, 3H), 1,689 (s, 3H), 1,871 (s, 3H), 2,209 (s, 3H), 2,450 (s, 3H), 3,800 (d, H) 4,119 (d, H) 4,305 (d, H) 4,413 (m, H) 4,563 (d, H) 4,703 (d, H) 4,940 (d, H), 4,958 (DD, H) 5,667 (d, H) 5,822 (DD, H), 6,263 (m, 2H), 7,019 (d, NH), 7,293 - 8,127 (m, 15H).

Mass spectrum: [M+H]+, 914, [M+Na]+, 936; [M+K]+, 952.

The mass spectrum of the high-resolution: MH+- 914,3394 (calculated 914,3422)

(c) Obtaining 2'-O-(dibenzylideneacetone)paclite is oratane (12 ml) was added powdered molecular sieves (1.0 g), and then was added dropwise a solution of a mixture of N-iodosuccinimide (0.33 g, 1,4622 mmol) and dibenzylamine (0,41 g, 1,4622 mmol) in dry tetrahydrofuran (8 ml). The resulting mixture was stirred at room temperature for 1 hour, then filtered through celite. The filtrate was evaporated to dryness, and the red residue was dissolved in ethyl acetate (50 ml) and washed with cold 6% NaHSO4cold 6% NaHCO3the brine. The solution was dried (MgSO4) and evaporated, resulting in the foam, it was purified by coated silica gel plates (methylene chloride-20% acetonitrile) and obtained the pure product (0,77 g, 69%).

IR-spectrum: 3854, 3744, 3362, 3066, 1960, 1722, 1602, 1580 cm-1.< / BR>
NMR (CDCl3): 1,075 (s, 3H), 1,167 (s, 3H), 1,651 (s, 3H), 1,799 (s, 3H), 2,209 (s, 3H), 2,296 (s, 3H), 2,464 (m, H) 3,686 (m, H) 4,121 (d, H) 4,240 (d, H) 4,293 (m, H), 4,808-4,957 (m, 6H), 5,006 (m, H), 5,565-5,649 (m, 2H), 6,034 (t, H) 6,194 (3, H), 7,100-8,132 (m, 26H).

Mass spectrum: [M+H]+, 1144; [M+Na]+, 1166; [M+K]+, 1182.

(d) Obtaining 2'-O-(phosphonomethyl)of paclitaxel

< / BR>
The mixture obtained in stage (C) product (0.9 g, 0,7874 mmol) and 10% PD/C (1 g) in ethyl acetate (20 ml) was hydrogenosomal at 60 psi (400 kPa) for 24 hours. The reaction mixture was filtered through celite, and the filtrate was evaporated to dryness. Ostanovka product (0,254 g, 33.4 per cent);

so pl. 202 - 205oC (decomposition).

IR-spectrum (KBr): 3438, 3066, 2942, 1722, 1652, 1602 cm-1.

NMR (acetone-d6/D2O): 1,081 (s, 6H) 1,571 (s, 3H), 1,847 (s, 3H), 2,115 (s, 3H), 2,357 (s, 3H), 3,707 (d, H) 4,08 (m, 2H), 4,275 (m, H), 4,941 - 5,085 (m, 4H), 5,231 (t, H) 5,430 (d, H) 5,544 (d, H) 5,970 (t, H) 6,376 (m, H) 6,961 - 8,017 (m, 16H).

Mass spectrum: [M+Na]+, 986, [M+K]+, 1002; [M+2Na-H]+, 1008; [M+Na+K-H]+, 1024; [M+2K-H]+, 1040.

The mass spectrum of the high-resolution: MNa+- 986,2955 (calculated 986,2976).

Example 5. 2',7-O-(Phosphonomethyl)of paclitaxel sodium salt

(a) Obtaining 2',7-O-bis(methylthiomethyl)of paclitaxel

< / BR>
To a stirred solution of paclitaxel (0,853 g, 1 mmol) and dimethyl sulfide (1,465 g, 20 mol) in acetonitrile (20 ml) at 0oC was added solid benzoyl peroxide (1,995 g, 8 mmol). The reaction mixture was intensively stirred at 0oC for 3 hours. The reaction course was monitored by thin-layer chromatography in a mixture of hexane:ethyl acetate (1:1, volume/volume), Rfpaclitaxel = 0,24, Rfproduct = 0,60. After the disappearance of the starting material (approximately 3 hours) the reaction was quickly stopped by evaporation of solvent to dryness at 25oC using a vacuum line in the room. Dry OST the 4 inch (19 mm), the solvent system: hexane: ethyl acetate (3:2, vol/vol), the volume of each fraction: approximately 25 ml Of fractions 15 to 19 was extracted specified in the header connection (0,515 g, 53% yield).

Mass spectrum (FAB/NOBA matrix, NaI, KI): [M+H]+m/z 974; [M+Na]+m/z 996; [M+K]+m/z 1012.

UV spectrum (MeOH):max= 204 nm, E (1%/1 cm)= 243,45;max= 228 nm, E (1%/ 1 cm) = 313,99.

IR-spectrum (KBr): 3440, 3064, 2926, 1724, 1668, 1602, 1582, 1514, 1484, 1452, 1372, 1314, 1266, 1242, 1178, 1142, 1068, 1026, 990, 915, 886, 848, 800, 774, 710, 646, 606, 570, 540, 480 cm-1.

1H NMR (CDCl3), : 1,17 (3H, s) of 1.20 (3H, s), by 1.68 (3H, s), of 1.74 (3H, s), 1,84 (H, DD), 2,04 (3H, d), is 2.09 (3H, s), of 2.15 (3H, s) overlaps with (H, m), 2,37 (H, DD), of 2.51 (3H, s), 2,79 (H, DDD), 3,78 (H, d), 4,18 (H, d), to 4.28 (H, m), or 4.31 (H, d), 4.53-in - 4,74 (4H, two overlapping AB m), is 4.93 (H, d), 4.95 points (H, d), 5,68 (H, d), of 5.82 (H, DD), 6,24 (H, DD), 6,54 (H, s), 7,05 (H, d), 7,28 - to 7.59 (10H, overlapping m), EUR 7.57 (H, m), 7,76 (2H, d), of 8.09 (2H, d).

(b) Obtaining 2',7-O-(dibenzylideneacetone)of paclitaxel

< / BR>
To a mixture of 2', 7-O-bis(methylthiomethyl)paclitaxel (198 mg, 0.2 mmol) and molecular sieves (approximately 200 mg) in methylene chloride (12 ml) at room temperature was added a solution of N-iodosuccinimide (135 mg, 0.5 mmol) and dibenzylamine (167 mg, 0.5 mmol) in dry tetrahydrofuran (8 ml). The reaction mixture was stirred 1.5 hours and dry at room temperature using a household vacuum line. The residue was dissolved in ethyl acetate (100 ml) and washed in a separating funnel with 1% sodium thiosulfate (50 ml), 0.5 m sodium bicarbonate (50 ml) and twice with water (2 x 50 ml). The organic phase was dried over magnesium sulfate, evaporated to dryness and re-dissolved in ethyl acetate (1 ml). The product was besieged by 50 ml of a mixture of diethyl ether: hexane (1:1) and twice washed with the same solvent system (2 x 50 ml). Obtained crude product (218 mg) with 74% yield. Purification of this product was carried out by loading its solution in methylene chloride (3 ml) on silica gel (f = 3/4 inch (19 mm) x L = 1 inch (25.4 mm) and elution of the product with 50 ml of a mixture of methylene chloride: ethyl acetate (3: 1). Got mentioned in the title compound (172,7 mg) with a yield of 59.3%.

Mass spectrum (FAB, NOBA matrix/NaI, KI): [M + Na]+m/z 1456; [M + K]+m/z 1472.

UV spectrum (MeCN): max= 194 nm, E (1%/1 cm) = 1078,36;max= 228 nm, E (1%/1 cm) = 311,95.

IR-spectrum (KBr): 3430, 3066, 3032, 2958, 1744, 1726, 1664, 1602, 1582, 1532, 1488, 1456, 1372, 1270, 1244, 1158, 1108, 1068, 1016, 1000, 952, 886, 800, 776, 738, 698, 604, 498 cm-1.

1H-NMR (CDCl3), : 1,12 (3H, s) to 1.14 (3H, s), and 1.56 (H, m), rate of 1.67 (3H, s) of 1.84 (3H, d), 1,90 (H, m), 2,17 (3H, s) to 2.29 (3H, s), 2,73 (H, m), of 3.73 (H, d), 4,08 (H, d), 4,15 (H, m), 4,20 (H, d), 4,77 (H, m), 4,79 (H, d), 4,91 - 5,04 (10H, overlapping m), 5.25-inch (H, DD), 5,38 (H, DD), 5,54 - 5, d).

(c) obtaining the sodium salt of 2',7-O-bis(phosphonomethyl)of paclitaxel

< / BR>
A sample of 2', 7-O-bis(dibenzylideneacetone)paclitaxel (112 mg, 0,078 mmol) was dissolved in ethyl acetate (7 ml) and was hydrogenosomal 10% palladium on coal (50 mg) at room temperature and 60 psi (400 kPa) for 2 hours. The catalyst was removed by filtration on celite. Celite was washed with ethyl acetate (10 ml). The filtrate was treated with solid sodium bicarbonate (20 mg, 3 equiv.), then the solvent evaporated to dryness. The dry residue was again dissolved in 5 ml of a mixture water: acetone (4:1, vol/vol) and was purified by chromatography on a column of C-18 reverse phase (55 - 105 MK C-18, waters, 50 ml of dry C-18, f = 3/4 inch (19 mm) in a mixture of water: acetone (4: 1, volume/volume). Eluent on an analytical column (C-18 Jones for PL C (15 cm, 1 ml/min, = 230 nm) in buffer acetonitrile: phosphate pH 6 (50/50, vol/vol) supplemented with a cocktail Q12 of ion pairs (Regis), Rt= 4.7 minutes. The fractions containing specified in the header of the product were combined, evaporated acetone under household vacuum at 20oC, and the solution liofilizirovanny. Was obtained is listed in title product (44,2 mg) with a yield of 58.8%.

Mass spectrum (FAB, NOBA matrix/NaI, KI): [M + H]+m/z 1118; [M + Na]+m/z 1140.

066, 2956, 1724, 1658, 1604, 1582, 1520, 1486, 1452, 1374, 1316, 1256, 1152, 1110, 1070, 1026, 966, 914, 802, 772, 710, 538 cm-1.

1H-NMR (acetone-d6/D2O): 0,97 (3H, s) of 1.02 (3H, s), 1,47 (H, m), and 1.54 (3H, s), 1.70 to (H, m), of 1.75 (3H, s), 1,85 (H, m), 2,11 (3H, s), is 2.30 (3H, s), 2,88 (H, m), 3,64 (H, d), a 4.03 (H, m) 4,06 (H, d), 4,16 (H, d), 4,74 (H, m), a 4.86 (H, m), 51,11 (H, PC), 5,22 (H, d), 5,42 (H, d), 5,90 (H, PC), 6,21 (H, s), 7,06 (H, PC), 7,32 - 7,69 (10H, overlapping m), 7,80 (2H, d), to 7.93 (2H, d).

Example 6. 7-O-Methylthiopyrimidin III

< / BR>
To a solution of 2'-O-ethoxycarbonyl-7-O-methylthiotetrazole (compound of example 3 (b), 27 g, a 27.4 mmol) in 100 ml of tetrahydrofuran and 500 ml of methanol was added svejeispechennyi K2CO3(2.7 g, 19 mmol). The solution was stirred for 30 minutes and neutralized resin IP-120 (H+), filtered and concentrated. Then the crude filtrate was dissolved in 200 ml dichloromethane and stirred for 24 hours with tetrabutylammonium (10 g). The solution was diluted with dichloromethane and washed with water, saturated bicarbonate solution and brine. After that, the organic fraction was dried over MgSO4and concentrated. The residue was chromatographically on silica gel (1:1, hexane:ethyl acetate) and obtained 9.4 g specified in the connection header (53%) with a melting point of 269oC.

Mass spectrum FAB (NOBA) M + H calc. DL-1
.

1H-NMR (CDCl3, 300 MHz): 8,08 (d, J = 7,1 Hz, 2H), 7,58 (t, J = 7.5 Hz, 1H), 7,45 (t, J = 7.8 Hz, 2H), 6,55 (s, 1H), 4,94 (d, J = 8,1 Hz, 1H), a 4.83 (Shir. square, J = 5,1 Hz, 1H), 4,66 (Avcv, J = 14,7 and 12.3 Hz, 2H), 4,30 (m, 2H), 4,13 (d, J = 8,4 Hz, 1H), 3,91 (d, J = 6,6 Hz, 1H), and 2.79 (m, 1H), and 2.27 (s, 3H), of 2.25 (m, 2H), 2,19 (s, 3H), of 2.16 (s, 3H), 2,10 (s, 4H), is 1.81 (m, 1H), 1,72 (s, 3H), of 1.61 (m, 2H), 1,16 (s, 3H), of 1.03 (s, 3H).

13C NMR (CDCl3, and 75.5 Hz), : 202,3, 170,8, 169,3, 167,0, 144,2, 132,6, 132,1, 130,1, 129,4, 128,6, 83,9, 80,9, 78,7, 75,7, 74,5, 73,9, 67,9, 57,6, 47,6, 42,7, 38,3, 26,7, 22,6, 21,0, 20,1, 15,2, 15,0, 10,8.

Example 7. Triethanolamine salt of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3' -(2-furyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine

(a) Obtaining 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2-furyl)-7-O - methyldiethanolamine

< / BR>
To a solution of hexamethyldisilazane (and 0.40 ml, 1,90 mmol) in 15 ml of tetrahydrofuran was added a solution of n-utility (0.75 ml, 2.5 M in hexano, 1.88 mmol) and stirred 5 minutes at -55oC. To this solution was added 7-methylthiopyrimidin III (compound of example 6, of 1.03 g of 1.59 mmol) in 10 ml of tetrahydrofuran and stirred for 10 minutes before adding 10 ml of a solution of (3R, 4R)-1-(tert-butyloxycarbonyl)-4-(2-furyl)- 3-(triethylsilyl)-2-azetidinone (883 mg, of 2.40 mmol). Removed a cool bath, and replaced her bath at 0oa solution of NH4Cl, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (2,5:1, hexane:ethyl acetate) and received 1.5 g of the reaction product of a combination of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'- (2-furyl)-7-O-methylthiomethyl-2'-O-criticalimportance (93%).

Mass spectrum FAB /NOBA/ [M+Na] calc. for C50H71NSSiO16:1036 found: 1036.

IR spectrum (film): 3446 (), 1720, 1368, 1242, 1166, 1144, 1124, 1066 cm-1.

1H NMR (CDCl3, 300 MHz): 8,07 (d, J = 7.2 Hz, 2H), 7,56 (m, 1H), 7,46 (t, J = 7.5 Hz, 2H), was 7.36 (m, 1H), 6,56 (s, 1H), 6,33 (m, 1H), 6,20 (m, 2H), 5,67 (d, J = 6,9 Hz, 1H), from 5.29 (CL, 2H), 4,94 (d, J = 7.8 Hz, 1H), 4.75 in (C, 1H) and 4.65 (s, 2H), 4,28 (m, 2H), 4.16 the (d, J = 8,1 Hz, 1H), 3,89 (d, J = 6,9 Hz, 1H), 2,80 (m, 1H), 2,46 (s, 3H), is 2.37 (m, 1H), 2,22 (m, 1H), 2,16 (s, 3H), 2,10 (s, 3H), 2,04 (s, 3H), of 1.84 (m, 1H), of 1.74 (s, 3H), of 1.65 (m, 1H), 1,33 (s, 9H), of 1.20 (s, 3H), 1,19 (s, 3H), 0,81 (t, J = 7.8 Hz, 9H), 0,47 (m, 6H).

13C NMR (CDCl3; 75,5 Hz), : 202,0, 171,2, 170,3, 169,3, 167,1, 155,3, 152,0, 141,9, 141,0, 133,6, 132,9, 130,2, 129,2, 128,7, 110,7, 107,3, 84,0, 81,1, 80,2, 78,7, 76,1, 75,7, 74,7, 74,1, 72,4, 71,1, 57,4, 52,8, 47,1, 43,3, 35,2, 33,0, 28,1, 26,3, 22,9, 21,2, 21,0, 15,0, 14,5, 10,9, 6,5, 4,3.

To the solution obtained above 2'-triethylsilyl ester (330 mg, 0.32 mmol) in 7 ml of tetrahydrofuran was added tetrabutylammonium (0,35 ml, 1.0 M THF, 0.35 mmol) and stirred 10 minutes. The solution was diluted with ethyl acetate and probyval:ethyl acetate) and received in the 301 mg specified in the connection header (95%).

Mass spectrum FAB (NOBA) M+H calc. for C45H58NO16S: 900. Found: 900.

IR spectrum (film): 3442, 1720, 1242, 1066, 1026 cm-1.

1H NMR (CDCl3, 300 MHz): 8,07 (d, J = 7,3 Hz, 2H), EUR 7.57 (t, J = 7,3 Hz, 1H), 7,45 (t, J = 7.8 Hz, 2H), 7,38 (s, 1H), 6,53 (s, 1H), 6,34 (d, J = 3.2 Hz, 1H), 6,29 (d, J = 3.2 Hz, 1H), 6,17 (t, J = 8,1 Hz, 1H), 5,65 (d, J = 6,9 Hz, 1H), from 5.29 (m, 2H), 4.92 in (d, J = 8.0 Hz, 1H), 4,70 (m, 1H), with 4.64 (d, J = 4,6 Hz, 2H), 4,29 (m, 2H), 4,14 (d, J = 8,3 Hz, 1H), 3,86 (d, J = 6,8 Hz, 1H), 3,37 (d, J = 5.8 Hz, 1H), 2,77 (m, 1H), of 2.38 (s, 3H), 2,32 (m, 2H), 2,16 (s, 3H), 2,10 (s, 3H), 2,02 (s, 3H), 1.77 in (m, 3H) 1,73 (s, 3H), of 1.33 (s, 9H), of 1.17 (s, 3H), of 1.12 (s, 3H).

13C NMR (CDCl3, and 75.5 Hz), 202,0, 172,6, 170,3, 169,2, 167,0, 155,2, 151,3, 142,4, 140,4, 133,7, 133,2, 130,2, 129,1, 128,7, 110,7, 107,4, 83,9, 81,2, 80,5, 78,6, 76,5, 76,1, 75,4, 74,6, 74,0, 72,5, 71,8, 57,4, 51,7, 47,2, 43,2, 35,2, 32,8, 28,1, 26,4, 22,6, 20,9, 15,2, 14,6, 10,9, 8,3.

(b) Obtaining 3'-N-desbenzoyl-3'-destiny-3'-N-(tert - butyloxycarbonyl)-3'-(2-furyl)-2'-O-ethoxycarbonyl-7-O - methyldiethanolamine

< / BR>
To the solution obtained in stage (a) product (864 mg, 0.96 mmol) in 50 ml of dichloromethane at 0oC was added diisopropylethylamine (2.0 ml, 11.5 mmol) and ethylchloride (0,50 ml of 5.25 mmol) and stirred 4 hours. The solution was diluted with dichloromethane and washed with saturated solution of bicarbonate, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (1:pectr FAB (NOBA) M+H calc. for C48H62NO18S: 972,3688. Found: 972,3654.

IR spectrum (film): 1752, 1720, 1370, 1244, 1196, 1176, 1064 cm-1.

1H NMR (CDCl3, 300 MHz): of 8.09 (d, J = 7.8 Hz, 2H), EUR 7.57 (t, J = 7.5 Hz, 1H), 7,46 (t, J = 7.8 Hz, 2H), 7,38 (s, 1H), 6,55 (s, 1H), 6.35mm (m, 1H), 6,27 (m, 1H), from 6.22 (t, J = 7.8 Hz, 1H), 5,67 (d, J = 7.2 Hz, 1H), 5,51 (d, J = 9.9 Hz, 1H), of 5.34 (d, J = 2.4 Hz, 1H), 5.25-inch (d, J = 10,2 Hz, 1H), 4.95 points (d, J = 8,1 Hz, 1H) and 4.65 (s, 2H), 4,30 (m, 2H), 4,22 (m, 2H), 3,88 (d, J = 7.2 Hz, 1H), 2,81 (m, 1H), 2,41 (s, 3H), 2,36-of 2.21 (m, 2H), of 2.16 (s, 3H), 2,11 (s, 3H), of 2.09 (s, 3H) and 1.83 (m, 1H), 1,74 (s, 3H), 1,67 (s, 1H), 1,59 (s, 1H), of 1.34 (s, 9H), of 1.29 (t, J = 7.2 Hz, 3H), of 1.20 (s, 3H), of 1.18 (s, 3H).

13C NMR (CDCl3, and 75.5 Hz), : 202,1, 169,9, 169,1, 167,6, 167,0, 154,0, 150,1, 142,6, 141,0, 133,6, 132,9, 130,2, 129,2, 128,7, 110,7, 107,5, 83,9, 81,1, 80,7, 78,7, 76,0, 75,7, 75,1, 74,7, 74,2, 71,8, 65,1, 57,4, 49,7, 47,1, 43,2, 35,0, 33,0, 28,1, 26,3, 22,6, 21,1, 20,9, 15,1, 14,5, 14,1, 10,9.

(c) Obtaining 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2-furyl)- 2-'-O-ethoxycarbonyl-7-O-dibenzylethylenediamine

< / BR>
To the solution obtained in stage (b) product (230 mg, 0,236 mmol) in 10 ml of anhydrous tetrahydrofuran was added 300 mg of molecular sieves, dipentylester (270 mg, 0.98 mmol) and recrystallized NIS (62 mg, 0.28 mmol). To this solution was added triftorbyenzola silver (45 mg, 0,17 mmol) and the solution was stirred for 3 hours. The solution was filtered through celite and razbam, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (15% acetonitrile in chloroform) and received 219 mg dimensiondata specified in the title compound (77%).

Mass spectrum FAB (NOBA) calc. for C61H72NPO22Na: 1224. Found: 1224.

IR spectrum (film): 3422 (Shir. ), 1750, 1722, 1370, 1244, 1160, 1036, 1016, 1000, 976, 944 cm-1.

1H NMR (CDCl3, 300 MHz): 8,08 (d, J = 6,9 Hz, 2H); 7,58 (t, J = 7.2 Hz, 1H), 7,46 (t, J = 7.8 Hz, 2H), 7,39 (s, 1H), 7,31 (m, 1H), 6.35mm (m, 2H), 6,28 (s, 1H), 6,21 (t, J =7.8 Hz, 1H), 5,64 (d, J = 6,9 Hz, 1H), 5,50 (d, J = 10.5 Hz, 1H), 5,39 (d, J = 6,6 Hz, 1H), 5,32 (d, J = 2.4 Hz, 1H), 5.25-inch (d, J = 9.9 Hz, 1H), free 5.01 (DD, J = 8.1 and 6.3 Hz, 5H), a 4.86 (d, J = 8,4 Hz, 1H), 4,29 - 4.09 to (m, 4H), 3,85 (d, J = 6,9 Hz, 1H), 2,77 (m, 1H), is 2.40 (s, 3H), 2,30 (m, 2H), 2,16 (s, 3H), of 1.99 (s, 3H), of 1.94 (m, 1H), 1,70 (s, 3H), 1,67 (s, 1H), and 1.54 (s, 1H), of 1.34 (s, 9H), of 1.28 (t, J = 7.2 Hz, 3H), of 1.20 (s, 3H), of 1.17 (s, 3H).

13C NMR (CDCl3, and 75.5 Hz), : 201,8, 169,9, 169,2, 167,7, 167,0, 155,1, 154,0, 150,0, 142,74, 141,1, 133,7, 132,9, 130,2, 129,1, 128,7, 128,5, 128,4, 128,0, 110,7, 107,6, 93,8, 84,1, 81,6, 80,8, 80,7, 78,8, 76,3, 75,1, 74,6, 71,8, 69,3, 69,2, 65,1, 57,0, 49,7, 46,7, 43,2, 35,0, 28,1, 26,4, 22,6, 21,2, 20,8, 14,6, 14,1, 10,5.

(d) Receiving triethanolamine salt of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'- (2-furyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine

< / BR>
To the solution obtained in stage (c) product (311 mg, 0,259 mmol) in is. Remove the catalyst by filtration through celite, and the filtrate was concentrated in vacuum. The residue was dissolved in 3 ml of ethyl acetate, was added triethanolamine (2.3 ml, 0.1 M in utilizethe, 0.23 mmol). The solution was concentrated and the residue was chromatographically on C18 (40% acetonitrile in water) and liofilizirovanny, resulting in 205 mg triethanolamine salt (67%) phosphonic acid.

Mass spectrum FAB (NOBA): M + Na calc. for C47H60HPO22Na: 1044. Found: 1044.

IR spectrum (film): 3432 (Shir. ), 1752, 1722, 1372, 1246, 1158, 1108, 1096, 1070, 1002 cm-1.

1H NMR (d6acetone/D2O, 300 MHz): of 8.09 (d, J = 7.2 Hz, 2H), 7.62mm (m, 2H), 7,52 (t, J = 7.5 Hz, 2H), 6.48 in (d, J = 3.3 Hz, 1H), 6.42 per (m, 2H), 6,16 (t, J = 8.7 Hz, 1H), 5,65 (d, J = 6,9 Hz, 1H), 5,46 (d, J = 3.6 Hz, 1H), and 5.30 (d, J = 3.6 Hz, 1H), 5,17 (SHS, 1H), 5,01 (sm, J = 9.0 Hz, 1H), 4,19 (SHS, 1H), 4,18 (m, 5H), 3,95 (m, 6H), a 3.87 (d, J = 6,9 Hz, 1H), 3,68 (s, 7H), 3,50 (PC, J = 4,8 Hz, 6H), 2.95 and (m, 1H), 2,44 (s, 3H), 2,41 (m, 2H), of 2.16 (s, 3H), and 1.9 (s, 3H), of 1.94 (m, 1H), 1,68 (s, 3H), of 1.34 (s, 9H), 1,24 (t, J = 6.9 Hz, 3H), of 1.17 (s, 6H).

Example 8. Triethanolamine salt of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'- (2-thienyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine

(a) Obtaining 3'-N-desbenzoyl-3'-destiny-3'-N-(tert - butyloxycarbonyl)-3'-(2-thienyl)-7-O-methyldiethanolamine

< / BR>
To a solution of the chickpeas added dropwise 7-methylthiopyrimidin III (1,15 g, 1.78 mmol) in 18 ml of THF and stirred in the cold for 10 minutes. Added ()CIS-1- (tert-butyloxycarbonyl)-4-(2-thienyl)-3-(triethylsilyl)- 2-azetidinone (2,80 g, 7,3 mmol) in 18 ml of THF, and allowed the cold water to rise slowly to 0oC for 30 minutes. The solution was diluted with ethyl acetate, washed with saturated solution of NH4Cl, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (5 : 1, hexane : ethyl acetate), obtaining of 1.87 g of recovered lactam (3 : 1, hexane : ethyl acetate) which gave 1.44 g of the reaction product of a combination of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'- (2-thienyl)-7-O-methylthiomethyl-2'-O-criticalimportance (78%).

Mass spectrum FAB (NOBA): M + Na calc. for C51H71NO15S2SiNa: 1052. Found: 1052.

IR spectrum (film): 3442 (Shir. ), 1720, 1490, 1368, 1270, 1242, 1162, 1110, 1064, 1024, 984, 754 cm-1.

1H NMR (CDCl3, 300 MHz): of 8.09 (d, J = 7.2 Hz, 2H), EUR 7.57 (t, J = 7,6 Hz, 1H), 7,47 (t, J = 7.8 Hz, 2H), 7,22 (m, 1H), 6,95 (m, 2H), 6,55 (s, 1H), 6,21 (t, J = 9,3 Hz, 1H), of 5.68 (d, J = 6,9 Hz, 1H), 5,49 (sm, 1H), 5,39 (sm, J = 9.6 Hz, 1H), 4,94 (d, J = 7.8 Hz, 1H) and 4.65 (s, 2H), 4,57 (s, 1H), 4,28 (m, 2H), 4,17 (d, J = 8,4 Hz, 1H), 3,88 (d, J = 6,9 Hz, 1H), 2,80 (m, 1H), 2,46 (s, 3H), is 2.37 (m, 1H), measuring 2.20 (m, 1H), 2,17 (s, 3H), of 2.10 (s, 3H), 2,03 (s, 3H), of 1.84 (m, 1H), 1,74 (s, 3H), 1,68 (s, 1H), 1,62 (s, 1H), 1,31 (s, 9H), of 1.20 (s, 6H), 030,2, 129,2, 128,7, 126,9, 124,6, 83,9, 81,2, 80,1, 78,8, 77,4, 76,0, 75,7, 75,2, 74,8, 74,1, 71,3, 57,4, 53,8, 47,0, 43,3, 35,3, 33,3, 28,1, 26,3, 23,0, 21,3, 20,9, 14,9, 14,4, 10,9, 6,6, 4,5.

To the solution obtained above 2'-triethylsilyl ether (1,41 g, 1.37 mmol) in 14 ml of tetrahydrofuran was added tetrabutylammonium (1,4 ml, 1.0 M in THF, of 1.40 mmol). The solution was stirred for 30 minutes, diluted with ethyl acetate, washed with brine, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (hexane : ethyl acetate, 1 : 1) and received 1,16 g specified in the title compound (92%).

Mass spectrum FAB (NOBA): M + Na calc. for C45H57NO15S2Na: 938. Found: 938.

IR spectrum (film): 3440 (Shir.), 1720, 1368, 1242, 1168, 1106, 1066, 710 cm-1.

1H NMR (CDCl3, 300 MHz): 8,08 (d, J = 7.2 Hz, 2H), to 7.59 (m, 1H), 7,47 (t, J = 7.8 Hz, 2H), 7,24 (m, 1H), 7,07 (m, 1H), 6,99 (m, 1H), 6,53 (s, 1H), 6,18 (t, J = 8,1 Hz, 1H), to 5.66 (d, J = 6,9 Hz, 1H), 5,49 (d, J = 9.6 Hz, 1H), 5,32 (d, J = 9.6 Hz, 1H), 4.92 in (d, J = 7.8 Hz, 1H), 4,63 (m, 3H), 4,28 (m, 2H), 4,15 (d, J = 8,4 Hz, 1H), 3,86 (d, J = 6,9 Hz, 1H), 2,80 (m, 1H), 3,47 (d, J = 5.4 Hz, 1H), 2,78 (m, 1H), a 2.36 (s, 3H), 2,34 ( , 2H), 2,17 (s, 3H), 2,10 (s, 3H), from 2.00 (s, 3H) and 1.83 (m, 1H), 1,74 (s, 3H), 1,72 (s, 1H), 1,61 (s, 1H), 1,33 (s, 9H), to 1.21 (s, 3H), of 1.18 (s, 3H).

13C NMR (CDCl3, 75.5 MHz), : 201,9, 172,3, 170,3, 169,2, 167,0, 154,0, 141,5, 140,2, 133,7, 133,3, 130,2, 129,1, 128,7, 127,0, 125,4, 125,4, 83,9, 81,3, 80,4, 78,6, 76,1, 75,4, 74,5, 74,0, 73,4, 72,5, 57,5, 52,8, Il)-3'-(2-thienyl)-2'-O-ethoxycarbonyl - 7-O-methyldiethanolamine

< / BR>
To the solution obtained in stage (a) product (621 mg, 0,677 mmol) in 35 ml of dichloromethane at 0oC was added diisopropylethylamine (1.20 ml, 6,89 mmol) and ethylchloride (0.35 ml, 3.7 mmol) and stirred 1 hour. Removed the cold bath, and the solution was stirred for 2 hours, diluted with dichloromethane, washed with a saturated solution of bicarbonate, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (hexane : ethyl acetate = 1 : 1) and received 528 mg specified in the title compound (79%).

Mass spectrum FAB (NOBA): M + Na calc. for C43H61NO17S2Na: 1010. Found: 1010.

IR spectrum (film): 3510, 3440, 1752, 1720, 1370, 1244, 1198, 1170, 1026, 988, 756 cm-1.

1H-NMR (CDCl3, 300 MHz): of 8.09 (d, J=7.2 Hz, 2H), 7,58 (m, 1H), of 7.48 (t, J= 7.8 Hz, 2H), 7,26 (m, 1H), 6,99 (, 2H), 6,55 (s, 1H), 6,23 (t, J=9.0 Hz, 1H), of 5.68 (d, J=6,9 Hz, 2H), 5,33 (d, J=9.9 Hz, 1H), 5.25 in (d, J=2.4 Hz, 1H), 4,94 (d, J=7.8 Hz, 1H) and 4.65 (s, 2H), 4,33 - 4,08 (m, 5H), 3,88 (d, J= 6,9 Hz, 1H), 2,80 (m, 1H), 2.40 a (s, 3H), 2.40 a - 2,20 (m, 2H), 2,16 (s, 3H), 2,11 (s, 3H), 2,07 (s, 3H) and 1.83 (m, 1H), of 1.74 (s, 3H), 1.69 in (s, 1H), 1,60 (s, 1H), 1,33 (s, 9H), is 1.31 (t, J=7.2 Hz, 3H), of 1.20 (s, 3H), 1,19 (s, 3H).

13C-NMR (CDCl3, and 75.5 Hz), : 202,0, 169,7, 169,1, 167,5, 167,1, 154,0, 140,9, 133,6, 132,9, 130,2, 129,2, 128,7, 127,2, 125,4, 125,3, 83,9, 81,2, 80,6, 78,8, 76,9, 76,0, 75,7, 74,7, 74,2, 72,8, 72,0, 65,2, 57,4, 50,9, 47,1, 43,3, 35,1, 33,0, 28,1, 26,4, 22,7, 21,2, 20,9, 15,1, 14,5, 14,1, 10,9�-7-O - dibenzylethylenediamine

< / BR>
To the solution obtained in stage (b) product (516 mg, 0,522 mmol) in 15 ml of anhydrous tetrahydrofuran was added 530 mg of molecular sieves dimensionful (576 mg, of 2.09 mmol) and recrystallized N-iodosuccinimide (136 mg, 0,604 mmol). To this solution was added triftorbyenzola silver (50 mg, 0,194 mmol) and the solution was stirred for 1 hour. The solution was filtered through celite, diluted with ethyl acetate, washed with 10% solution NaS2O8, a saturated solution of bicarbonate and brine, dried over MgSO4and concentrated. The residue was chromatographically on silica gel (15% acetonitrile in chloroform) and received 535 mg specified in the connection header (84%).

Mass spectrum FAB (NOBA): M+Na calc. for C16H72NO21PSNa: 1240. Found: 1240.

IR spectrum (film): 3424 (Shir.), 1750, 1722, 1370, 1244, 1016, 1000, 944 cm-1.

1H-NMR (CDCl3, 300 MHz): 8,08 (d, J=7,0 Hz, 2H), 7,58 (m, 1H), 7,47 (t, J=7.5 Hz, 2H), 7,28 (m, 11H), of 6.99 (m, 2H), 6,33 (s, 1H), from 6.22 (t, J=7.8 Hz, 1H), 5,66 (m, 2H), 5,39 (t, J=6,6 Hz, 1H), of 5.34 (d, J=12 Hz, 1H), with 5.22 (d, J= 2.4 Hz, 1H), free 5.01 (DD, J=8.1 and 6.0 Hz, 5H), a 4.86 (d, J=7.8 Hz, 1H), 4,29 - 4,08 (m, 5H), 3,85 (d, J=6,6 Hz, 1H), was 2.76 (m, 1H), 2,39 (s, 3H), 2,35 - to 2.18 (m, 2H), 2,16 (s, 3H) 1,97 (s, 4H), 1.69 in (C, 4H), of 1.33 (s, 9H), of 1.30 (t, J=7.2 Hz, 3H), of 1.20 (s, 3H), of 1.17 (s, 3H).

13C NMR (CDCl3, and 75.5 Hz), : 197,4, 165,4, 164,9, 163,3, , 2,3, 38,9, 30,7, 23,8, 22,0, 18,3, 17,0, 16,4, 10,3, 9,8, 6,2.

(d) Receiving triethanolamine salt of 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2 - thienyl)-2'-O-ethoxycarbonyl-7-O-phosphonomethylglycine

< / BR>
To the solution obtained in stage (c) product (512 mg, 0.42 mmol) in 30 ml of ethyl acetate was added 53 mg of palladium on coal (10%), and the solution was stirred in an environment of hydrogen for 3 hours. Remove the catalyst by filtration through celite, and the filtrate was concentrated in vacuum. The residue was dissolved in 2 ml of ethyl acetate, was added triethanolamine (4,0 ml of 0.1 M acetate, 0.40 mmol). The solution was concentrated, and the residue was chromatographically on C18 (40% acetonitrile in water) and liofilizirovanny, resulting in 280 mg triethanolamine salt (56%). Analysis by HPLC showed a purity of salt equal to 96%.

Mass spectrum FAB (NOBA): M+Na calc. for C47H60NO21PS:1060. Found: 1060.

IR-spectrum (KBr): 3422 (Shir.), 1750, 1720, 1372, 1246, 1162, 1096, 1068, 1000 cm-1.

1H NMR (d6acetone/D2O, 300 MHz): of 8.06 (d, J=7.2 Hz, 2H), 7,63 (t, J= 7.2 Hz, 1H), 7,52 (t, J=7.8 Hz, 2H), 7,38 (d, J=4,2 Hz, 1H), 7,16 (d, J=3.5 Hz, 1H), 7,01 (DD, J=5.1 and 3.6 Hz, 1H) 6,37 (s, 1H), 6,11 (t, J=8.7 Hz, 1H), 5,61 (d, J=6,9 Hz, 1H), ceiling of 5.60 (s, 1H), 5,26 (d, J=4.5 Hz, 1H), 5,14 (d, J=6,6 Hz, 1H), 5,00 (d, J=8,4 Hz, 1H), a 4.86 (DD, J=12,0 and 6.3 Hz, 1H), 4 of 1.66 (s, 3H), of 1.30 (s, 9H), of 1.23 (t, J=7.2 Hz, 3H) 1.14 in (s, 6H).

Example 9. 10-deacetyl-3'-N-desbenzoyl-3'-N-(tert - butyloxycarbonyl)-10-O-(phosphonomethyl)paclitaxel

(a) Receiving 10-deacetyl-10-O-benzyloxycarbonyl-7-O - triethylenemelamine III

< / BR>
In a dry flask in an argon atmosphere containing 7-O-triethylsilyl-10 - deacetylbaccatin III (2,093 g 3,177 mmol), was added dry tetrahydrofuran (30 ml) and cooled to -70oC. There was added dropwise a 1.6 M n-utility (2.38 ml, 3,81 mmol). After stirring for 15 minutes, was added dropwise benzylchloride (0,91 ml of 6.35 mmol). The resulting mixture was stirred for 3 hours with gradual warming to ambient temperature. The reaction was quickly stopped by 25 ml of a saturated solution of NH4Cl, the mixture was washed with brine and dried over MgSO4. Flash chromatography (silica gel, 30 to 45% ethyl acetate in hexane) gave 2.24 g (89%) indicated in the title compound as a white foam.

1H NMR (CDCl3, 300 MHz): 8,10 (d, J=8.0 Hz, 2H), 7,63 - 7,58 (m, 1H), 7,47 (t, J= 8.0 Hz, 2H), 7,41 - 7,26 (m, 5H), of 6.29 (s, 1H), 5,61 (d, J=7,0 Hz, 1H), 5,20 (sq J=12,2 Hz, 2H), 4,96 (d, J=9.0 Hz, 1H), 4,87 - 4,84 (m, 1H), 4,48 (DD, J=6,7 and 10.4 Hz, 1H), 4,30 (d, J=8.5 Hz, 1H), 4,14 (d, J=8.5 Hz, 1H), 3,84 (d, J=7,0 Hz, 1H), 2,58 - 2,48 (m, 1H), to 2.29 (m, 4H), of 2.20 (s, 3H), 2,03 (d, J= 5.0 Hz, 1H), 1,92 of - 1.83 (m, 1H), 1,68 (s, 3H), of 1.17 (s, 3H), 1.04 million is -3'-N-(tert-butyloxycarbonyl)-2'-7-bis-O - criticalimportance

< / BR>
In a dry flask containing obtained in stage (a) product (3.50 g, was 4.42 mmol), was added a small amount of toluene, after which the solution was concentrated under vacuum. The flask was placed in an argon atmosphere, was added 100 ml of dry tetrahydrofuran. The flask was cooled to -70oC, was added dropwise 1.0 M hexamethyldisilazide lithium (to 6.19 ml, to 6.19 mmol). After stirring for 20 minutes was added dropwise a solution of (3R,4S)-1-(tert-butyloxycarbonyl)-4-phenyl-3-triethylsilyl-2 - azetidinone (2.58 g, 7,07 mmol) in 10 ml dry THF. The reaction mixture was stirred 3.5 hours, gradually warming to ambient temperature. Then it was rapidly cooled to 70 ml of a saturated solution of NH4Cl, washed with brine and dried over MgSO4. Flash chromatography (silica gel, 5 to 15% ethyl acetate in hexano) gave 5,12 g (99%) indicated in the title compound as a white foam.

1H NMR (CDCl3, 300 MHz): 8,11 (d, J= 8.0 Hz, 2H), 7,60 - 7,58 (m, 1H), of 7.48 (t, J = 8.0 Hz, 2H), 7.24 to 7,26 (m, 10H), 6,32 - 6,26, (m, 2H), 5,69 (d, J = 7 Hz, 1H), 5,47 (sm, J = 9.7 Hz, 1H), 5,31 - 5,10 (m, 3H), 4,94 (d, J = 8.5 Hz, 1H), 4,56 (s, 1H), 4,46 (DD, J = 6,9 and 10.6 Hz, 1H), or 4.31 (d, J = 8,3 Hz, 1H), 4,17 (q, j = 8,3 Hz, 1H), 3,81 (d, J = 7,0 Hz, 1H), 2,53 (s, 3H), 2,48 is 2.33 (m, 1H), 2,22 - 2,17 (m, 1H), 2,09 (s, 3H), 1,95 is 1.86 (m, 1H), 1.70 to (s, 3H), of 1.65 (s, 1H), 1,52 (s, 1H), of 1.30 (s, 9H), 1,26 - 1,1 the et-butyloxycarbonyl)-7-O - criticalimportance

< / BR>
Obtained in stage (b) product (5,12 g, 4.40 mmol) was dissolved in 100 ml of ethyl acetate, transferred into a vessel Parra and placed under a protective layer of argon. There was added 10% palladium on coal (2.4 g) and the reaction mixture was placed on a hydrogenation installation Parra (55 pounds per square inch, i.e., a 3.87 kg/cm2for 8 hours. The reaction mixture was filtered through celite and concentrated. Flash chromatography (silica gel, 15% to 20% ethyl acetate in hexane) gave 3,24 g (79%) indicated in the title compound as a white foam. The hydrolysis of 2'-triethylsilyl group of the product obtained at stage Parra.

1H NMR (CDCl3, 300 MHz): 8,10 (d, J = 8.0 Hz, 2H), 7,63 - 7,58 (m, 1H), 7,49 (d, J = 8.0 Hz, 2H), 7,39 - 7,26 (m, 5H), 6,27 - of 6.17 (m, 1H), 5,64 (d, J = 7,2 Hz), 5,42 (d, J = 9.4 Hz, 1H), 5,28 - a 5.25 (m, 1H), 5,12 (s, 1H), 4.92 in (d, J = 8.6 Hz, 1H), 4,62 (SHS, 1H), to 4.38 - to 4.28 (m, 3H), 4,17 (d, J = 8.5 Hz, 1H), 3,85 (d, J = 6,7 1H), 3,36 (d, J = 5.3 Hz, 1H), 2,49 - to 2.40 (m, 1H), a 2.36 (s, 3H), 2,25 (sm, J = 8.7 Hz, 2H), 1,99 is 1.91 (m, 1H), 1,85 (s, 3H), of 1.74 (s, 3H), 1.69 in (s, 1H), 1,67 (s, 1H), of 1.35 (s, 9H), 1,22 (s, 3H), 1,11 (s, 3H), of 0.93 (t, J = 7.5 Hz, 9H), and 0.61 - 0,49 (m, 6H).

(d) Receiving 10-desacetyl-2'-O-benzyloxycarbonyl-3'-N-desbenzoyl-3'-N-(tert - butyloxycarbonyl)-7-O-criticalimportance

< / BR>
Into the flask containing obtained in stage (C) the product (3,24 g, 3,51 mmol), was added 30 ml of dry dichloromethane. The flask PL), and then was added dropwise benzylchloride (1,00 ml, 7,02 mmol). After 15 minutes, cooled bath was removed and the reaction mixture was allowed to mix at ambient temperature for 7 hours. Then the mixture was rapidly cooled by 30 ml of a saturated solution of NH4Cl, washed with brine and dried over MgSO4. Flash chromatography (silica gel, 7 - 20% ethyl acetate in hexane) gave 3,24 g (89%) indicated in the title compound as a white solid.

1H NMR (CDCl3, 300 MHz): 8,10 (d, J = 8.0 Hz, 2H), 7,62 - EUR 7.57 (m, 1H), of 7.48 (t, J = 8.0 Hz, 2H), 7,40 - 7,26 (m, 10H), 6,33 - 6,27 (m, 1H), to 5.66 (d, J = 7,0 Hz, 1H), 5,49 - 5,42 (m, 2H), 5,31 (s, 1H), 5,22 - 5,13 (m, 3H), is 4.93 (d, J = 9.4 Hz, 1H), to 4.38 (DD, J = 6,5 and 10.7 Hz, 1H), 4,34 - to 4.28 (m, 2H), 4,18 (d, J = 8,3 Hz, 1H), 3,90 (d, J = 6,7 Hz, 1H), 2,52 - of 2.30 (m, 4H), 2,24 - of 2.20 (m, 1H), 1,97 - to 1.87 (m, 3H), of 1.74 (s, 3H), of 1.59 (s, 3H), 1,32 (s, 9H), 1.26 in (s, 3H), 1,11 (s, 3H), 0,96 - to 0.88 (m, 9H), and 0.61 - of 0.48 (m, 6H).

(e) Receiving 10-deazetil-2'-O-benzyloxycarbonyl-3'-N-desbenzoyl-3'-N-(tert - butyloxycarbonyl)-10-O-(dibenzylideneacetone)-7-O - criticalimportance

< / BR>
Obtained in stage (d) the product was dissolved in 13.5 ml (54%) DMSO, is 8.75 ml (35%) of acetic anhydride and 2.75 ml (11%) glacial acetic acid and placed in an atmosphere of argon. The reaction mixture was stirred for 56 hours, after which it was diluted is when the test indicator paper to determine the pH, and then washed with brine. The organic fraction was dried MgSO4and concentrated. Flash chromatography (15 - 20% ethyl acetate in hexane) to give 3.12 g of the crude white foam with the desired dimethylacetamide product (i.e., 10-desacetyl-2'-O-benzyloxycarbonyl-3'-N-desbenzoyl - 3'-N-(tert-butyloxycarbonyl)-10-O-(methylthiomethyl)-7-O - criticalimportance), constituting 70% of the material by NMR.

Then the above crude compound (3.12 g) was dissolved in 1,2-dichloroethane (61 ml) and placed under a protective layer of argon. Added powdered molecular sieves (3.12 g) and the resulting heterogeneous mixture was intensively stirred. Added by cannula a solution of recrystallized N-iodosuccinimide (0,830 g of 3.69 mmol) and dibenzylamine (of 1.027 g of 3.69 mmol) in dry tetrahydrofuran (46 ml). The resulting mixture was stirred 5 hours, filtered through a layer of celite and diluted to a volume of 250 ml of ethyl acetate. Washed it in cold 2% solution of NaHCO3(2 x 125 ml) cold 6% solution of NaHCO3(2 x 125 ml) and brine. The organic phase was dried (MgSO4) and concentrated. Flash chromatography (silica gel 25 - 35% ethyl acetate in hexane) gave 1.52 g (40%) indicated in the title compound as a white solid ve (t, J = 8.5 Hz, 1H), 5,65 (d, J = 6,8

Hz, 1H), 5,49 of 5.39 (m, 4H), 5,32 (s, 1H), 5,18 - 4,19 (m, 4H), is 4.93 (d, J = 9,2 Hz, 1H), of 4.44 (DD, J = 6,6 and 10.2 Hz, 1H), or 4.31 (d, J = 8,4 Hz, 1H), 4.16 the (d, J = 8.5 Hz, 1H), 3,80 (d, J = 6,9 Hz, 1H), 2,69 - 2,39 (m, 4H), 2,33 - of 2.23 (m, 3H), 2,03 (s, 3H), 1,90 (t, J = 12,6 Hz, 1H), 1,68 - to 1.63 (m, 6H), of 1.28 (s, 9H), 1,16 - of 1.10 (m, 6H), of 0.93 (t, J = 7.4 Hz, 9H), 0.55 and (sq, J = 7.8 Hz, 6H).

13C NMR (CDCl3, 75.5 MHz), : 204,1, 169,7, 167,9, 167,1, 151,1, 140,7, 135,7, 133,6, 130,2, 129,2, 128,9, 128,8, 128,7, 128,6, 128,5, 128,4, 128,3, 128,2, 128,0, 127,8, 126,4, 90,4, 84,2, 81,1, 80,4, 79,3, 78,8, 74,9, 72,8, 72,0, 70,5, 69,2, 69,1, 69,0, 58,1, 46,8, 43,2, 37,1, 35,0, 28,1, 26,5, 22,8, 21,0, 14,1, 10,0, 6,9, 5,5.

Mass spectrum (FAB): m/z+: 1345.

(f) Receiving 10-desacetyl-2'-O-benzyloxycarbonyl-3'-N-desbenzoyl-3'-N-(tert-butyloxycarbonyl)-10-O-(dibenzylideneacetone)of paclitaxel

< / BR>
The solution obtained in stage (e) product (50,8 mg of 0.038 mmol) in dry tetrahydrofuran (2.5 ml) under argon was cooled to -40oC. To this solution was added dropwise tetrabutylammonium (0,057 ml 0,057 mmol) in tetrahydrofuran (1.0 M). The reaction mixture was stirred for 1.5 hours while gradually heating to -20oC. the Mixture was rapidly cooled to 15 ml of a saturated solution of NH4Cl and diluted with 30 ml of ethyl acetate. The organic phase is washed with NaHCO3(2 x 15 ml) and brine. Then it was dried (MgSO4) and concentrated. Preparative tonkas the ideal of white powder.

1H NMR (CDCl3; 300 MHz): 8,10 (d, J = 8.5 Hz, 2H), 7,60 - of 7.55 (m, 1H), 7,49 - 7,44 (m, 2H), was 7.36 - to 7.18 (m, 20H), 6,27 - to 6.22 (m, 1H), 5,78 (s, 1H), 5,67 (d, J = 7,0 Hz, 1H), 5,44 - of 5.34 (m, 3H), 5,27 (d, J = 2.2 Hz, 1H), 5,24 - of 5.05 (m, 4H), 5,01 - 4,91 (m, 4H), 4,39 - to 4.28 (m, 2H), 4,17 (d, J = 8,2 Hz, 1H), a 3.87 (d, J = 7,0 Hz, 1H), 2,58 is 2.51 (m, 1H), 2,41 (s, 3H), 2.40 a - to 2.18 (m, 2H), 2.00 in to 1.87 (m, 5H), 1,73 was 1.69 (m, 4H), of 1.30 (s, 9H), 1,22 - of 1.15 (m, 6H).

Mass spectrum (FAB): m/z+: 1231.

(g) Receiving 10-deacetyl-3'-N-desbenzoyl-3'-N-(tert-butyloxycarbonyl)-10-O -(phosphonomethyl)of paclitaxel triethanolamine salt

< / BR>
A Parr flask with a capacity of 500 ml was loaded 10-desacetyl-2'-O - benzyloxycarbonyl-3'-N-desbenzoyl-3'-N-(Tetra-butyloxycarbonyl)-10-O- (dibenzylideneacetone)paclitaxel (264,9 mg, 0,215 mmol) and ethyl acetate (20 ml). Then the bulb blew a stream of argon, was added 10% Pd/C (318 mg). The resulting mixture was placed on the installation Parra in the environment of hydrogen under a pressure of 55 psig (3,86 kg/cm2). The reaction was monitored by HPLC (70:30 buffer CH3CN/Q8, pH 6,0, 1,00 ml/min, column ZORBAX C-18, 25,0 cm = 230 nm), until then, until it became visible the source material (12.5 hours). The mixture was filtered through a layer of celite, which was washed with ethyl acetate and a small amount of dichloromethane. The obtained filtrate was concentrated, and the residue was dissolved in di the slots (purity, when determining by HPLC - 80%) as a white solid. This material was sent directly to the next stage.

Into the flask containing the above free acid (140 mg, 0,153 mmol) was added dichloromethane (10 ml). Then the resulting solution was treated 0,100 M solution of triethanolamine in ethyl acetate (1,16 ml, 0,116 mmol), the resulting solution became turbid. Added about 2 ml of hexane and the mixture was kept at -20oC all night. The precipitate was filtered through a funnel with a porous (4,0 - 5,5 µm) glass plate. The solid was collected and placed under vacuum for 4 hours, which gave to 69.9 mg (42%) indicated in the title triethanolamine salt in the form of a grey powder, which had a purity of 95 to 96% in the determination by HPLC-analysis. (TR= 2,05 min, the Buffer 70:30

CH3CN/Q8, pH 6,0, 1,00 ml/min, ZORBAX C-18, 25,0 cm = 230 nm).

1H NMR (d6-acetone/D2O, 300 MHz): 8,03 (d, J = 7,4 Hz, 2H), 7,65 (t, J = 7,3 Hz, 1H), 7,54 (t, J = 7,6 Hz, 2H), 7,42 - 7,33 (m, 5H), 7,21 (t, J = 7,0 Hz, 1H), 6,09 (t, J = 9.0 Hz, 1H), of 5.81 (s, 1H), 5,59 (d, J = 7,0 Hz, 1H), 5,12 (CL, 2H), is 4.93 (d, J = 8,4 Hz, 2H), 4,56 (d, J = 4.9 Hz, 1H), or 4.31 - 4.26 deaths (m, 1H), 4,11 (s, 2H), 3,41 - 3,37 (m, 6H), 2,42 of - 2.32 (m, 5H), 2,15 (SHS, 1H), of 1.97 (s, 3H), 1.77 in - of 1.64 (m, 2H), 1,58 (s, 3H), of 1.13 (s, 9H), 1,15 - of 1.07 (m, 6H).

13C NMR (d6-acetone, D2O, 75.5 MHz), : 171,6, 166,9, 156,6, 141,8, 135,1, 134,2, pectr high resolution: MNa+- 940,3142,

Calculated for C44H56NO18PNa = 940,3133.

Example 10. 2'-O-phosphonomethylglycine

(a) Obtaining 2'-O-(methylthiomethyl)-7-O-criticalimportance

< / BR>
To a solution of 7-O-criticalimportance (70.0 mg, 72.2 mmol), bis (methylthiomethyl)ester (90 mg, 72.2 mmol), molecular sieves (70 mg) and N-iodosuccinimide (160 mg, 72.2 mmol) in tetrahydrofuran (2.0 ml) at room temperature was added triflate silver (5.0 mg, of 19.5 mmol) and the resulting solution was stirred for 2 hours. Then the reaction mixture was diluted with ethyl acetate and filtered through a layer of celite. The filtrate was washed with saturated aqueous sodium bicarbonate solution and then with a mixture (1:1, vol/vol) saturated aqueous sodium bicarbonate solution and 5% aqueous sodium thiosulfate solution and finally brine. The organics were then dried over sodium sulfate and concentrated in vacuum. The residual oil was purified by flash chromatography (3: 1, hexane-ethyl acetate) and received as a result of 22.0 mg (29%) indicated in the title compound as a white solid.

1H NMR (CDCl3, 300 MHz): 8,12 - 7,20 (15H, m),? 7.04 baby mortality (1H, d, J = 8,9 Hz), 6,41 (1H, s), and 6.25 (1H, m), of 5.81 (1H, DD, J = 8,9 and 2.4 Hz), of 5.68 (1H, d, J = 7,0 Hz), is 4.93 (1H, d, J = 8.0 Hz), 4,79 (21H, d, J = 6.9 Hz), 2,48 - 1,13 (25H, m, incl. the singlets at 2,51, 2,13, 2,05, 2,01, 1,69, 1,19, 1,16), 0,98 - 0,85 (9H, m), 0,65 - of 0.50 (6H, m).

(b) Obtaining 2'-O-(dibenzylideneacetone)-7 - criticalimportance

< / BR>
To the solution obtained in stage (a) product (15 mg, 0,0141 mmol) and molecular sieves (15 mg) in THF (0.5 ml) at room temperature was added dimensionful (20.0 mg, 0,089 mmol) and then N-yodsanklai (4.2 mg, 0,0187 mmol) and the solution was stirred for 1 hour. At this point, the analysis of the reaction mixture by thin-layer chromatography showed the presence only of the source material. After this was added in three portions over 2 hours triplet silver (5.0 mg, 0,0189 mmol) and the reaction mixture was stirred for another 1 hour. Then the reaction mixture was diluted with ethyl acetate, and the resulting solution was filtered through a layer of celite. The filtrate was treated with a mixture (1: 1 vol/vol) saturated aqueous sodium bicarbonate solution and 5% aqueous sodium thiosulfate solution. The organic extract is then washed with brine, dried over sodium sulfate and concentrated in vacuum. The residual oil was purified by flash chromatography (1:1, hexane:ethyl acetate) and received 5.0 mg (33%) specified in the connection header.

1H NMR (CDCl3, 300 7 and 5.6 Hz), 4,43 (1H, DD, J = 10; 4 and 6.7 Hz), the 4.29 (1H, d, J = 8,3 Hz), 4,16 (1H, d, J = 8,3 Hz), of 3.78 (1H, d, J = 7,0 Hz), 2,60 - 1,13 (22H, m, incl. the singlets at 2,49, 2,15, 1,93, 1,66, 1,15, 1,13, 3H each), of 0.95 is 0.84 (9H, m), 0,63 - 0,45 (6H, m).

(c) Obtaining 2'-O-phosphonomethylglycine

< / BR>
Obtained in stage (b) the product was treated with tetrabutylammonium in accordance with the procedure described in example 9 (f), to remove the 7-O-triethylsilanol group. Resulting compound was subjected to catalytic hydrogenation in accordance with the procedure described in the preceding examples, and has been specified in the header of the connection.

Example II. 2'-O-phosphonomethylglycine (Alternate method)

(a) Obtaining 2'-O-criticalimportance

< / BR>
To a solution of paclitaxel (20,0 g, 0,0234 mol) and imidazole (3,59 g, 0,052 mol) in 150 ml of DMF (dimethylformamide) at 0oC) was added in portions of 2 ml for 20 min triethylsilane (6,0 ml, 0,053 mol). Then the reaction mixture was stirred at 0oC for 1 hour. Next, the mixture was diluted with ethyl acetate and saturated aqueous solution of ameriglide. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated in vacuum, having rezultata 21,07 g (yield 98%) of the target specified in the connection header in the form of unpainted white solid.

1H NMR (CDCl3, 300 MHz): 8,15 (2H, m), of 7.70 (2H, m), 7,65 - 7,30 (11H, m), to 7.15 (1H, d, J = 8,9 Hz), 6,30 (1H, s), and 6.25 (1H, m), 6,70 - 6,10 (2H, m), 4,94 (1H, d, J = 7.9 Hz), of 4.67 (1H, d, J = 2.0 Hz), and 4.40 (1H, m), the 4.29 (1H, d, J = 8,4 Hz), 4,18 (1H, d, J = 8,4 Hz), 3,81 (1H, d, J = 7,1 Hz), 2,65 - 1,10 (22H, including singlets at 2,55, 2,20, 1,88 1,69, 1,22, 1,13, 3H each).

(b) Obtaining 2'-O-triethylsilyl-7-O-benzyloxycarbonylglycine

< / BR>
To a solution of 2'-O-criticalimportance (22,3 g, and 24.1 mmol) in THF (250 ml), cooled to -50oC, was added dropwise within 10 minutes utility (1.6 M in hexano, 12.9 ml of 8.06 mmol). The resulting solution was stirred 20 min while maintaining the temperature between -50oC and -35oC. Then the reaction mixture was cooled to -50oC and added dropwise within 5 minutes benzylchloride (5.0 ml, to 29.8 mmol). The reaction mixture was stirred at -40oC for 30 minutes, then brought balance to the 0oC for about 30 minutes. Then the mixture was diluted with ethyl acetate and saturated aqueous ammonium chloride, and the obtained organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuum.1H NMR analysis of the crude mixture showed the presence of the desired 2'-O-triethylsilyl-7-O-basiliximab the products used in the next stage without further purification and then separation of the isomers. An analytical sample of the primary product 2'-O-triethylsilyl-7-O-benzyloxycarbonylglycine was purified by flash chromatography.

1H NMR (CDCl3, 300 MHz): to 8.12 (2H, m), 7,72 (1H, m), 7,65 - 7,27 (1H, d, J = 8,8 Hz), 6,41 (1H, m ), of 6.20 (1H, m), 5,72 - the 5.65 (2H, m), 5,52 (1H, m), of 5.24 (1H, d, J = 12.3 Hz), 5,16 (1H, d, J = 12.3 Hz), of 4.95 (1H, d, J = 8.7 Hz), 4,69 (1H, s), 4,35 (1H, d, J = 8,3 Hz), 4,25 (1H, d, J = 8,3 Hz), of 3.94 (1H, d, J = 6.8 Hz), 2,70 - 1,12 (22H, incl. the singlets at 2,54, 2,14, 2,01, 1,80, 1,20, 1,15, 3H each), 0,81 - to 0.73 (9H, m), 0.55 to 0,31 (6H, m).

(c) Obtain 7-O-benzyloxycarbonylglycine

< / BR>
To the solution obtained in stage (b) product (24,0 g, and 22.6 mmol) in acetonitrile (250 ml), cooled to 0oC, was added hydrochloric acid (6 N., 1.0 ml, 6.0 mmol). After 10 minutes, analysis by thin-layer chromatography (hexane: ethyl acetate, 1: 1) showed that the reaction is complete. The reaction mixture was diluted with saturated aqueous sodium bicarbonate and then with ethyl acetate and the organic layer was separated, washed with brine, dried with sodium sulfate and concentrated in vacuum. The residual oil was purified by flash chromatography (hexane:ethyl acetate, 1:3, then 1:1) and obtained as a result of 11.4 g (48% over two stages) specified in the connection header and 4.8 g (20%) 7-epigallocatechol.

(d) Obtaining 2'-O-(methylthiomethyl)-7-O - benzyloxycarbonylglycine

< / BR>
To a solution of 7-O-benzyloxycarbonylglycine (5.53 g, 5,71 mmol) of 1,1'-decimalisation ester (7.8 g, to 57.1 mmol), N-iodosuccinimide (6,35 g, 28.3 mmol) and kiln dried powdered molecular sieves (5.0 g) in THF (110 ml) at room temperature was added triflate silver (300 mg, at 1.17 mmol). Analysis by thin-layer chromatography (hexane:ethyl acetate, 1:1) reaction mixture after 20 minutes showed a conversion of about 40% of the source material in the higher fractions. Then added triplet silver (150 mg, 0,585 mmol) and monitored the reaction by thin-layer chromatography, which showed that after 30 minutes the reaction was completed by approximately 65%. The mixture was diluted with ethyl acetate (100 ml), filtered through a layer of celite, and the filtrate was poured into a separating funnel containing 20 ml of a saturated aqueous solution of sodium bicarbonate and 50 ml of 5% aqueous sodium thiosulfate solution. Organicsue oil was purified by flash chromatography (hexane:ethyl acetate, gradient elution from 4:1 to 3:2) and obtained 3.0 g (yield 54%) indicated in the title compound as a pale yellow solid.

1H NMR (CDCl3, 300 MHz): 8,10 (2H, m), 7,74 (2H, m), 7,66 - 7,25 (18H, m), 7,05 (1 H, d, J = 8,9 Hz), 6,40 (1H, s), of 6.26 (1H, m), 5,77 (1H, DD, J = 8.8 and 2.5 Hz), 5,71 (1H, d, J = 6.9 Hz), the 5.51 (1H, DD, J = 10.6 and a 7.1 Hz), to 5.21 (1H, d, J = 11,9 Hz), 5,14 (1H, d, J = 11,9 Hz) to 4.92 (2H, m), and 4.68 (1H, d, J = 2.5 Hz), or 4.31 (1H, d, J = 11.8 Hz), 4,30 (1H, d, J = 8.5 Hz), 4,16 (1H, d, J = 8.5 Hz), 4,10 (1H, d, J = 11.8 Hz), 3,93 (1H, d, J = 6.9 Hz), 2,65 - 1,10 (25H, incl. the singlets at 2,50, 2,15, 2,05, 1,74, 1,72, 1,20, 1,15, 3H each).

(e) Obtaining 2'-O-dibenzylideneacetone)-7-O - benzyloxycarbonylglycine

< / BR>
To a solution of 2'-O-(methylthiomethyl)-7-O-benzyloxycarbonylglycine (1.06 g, 1.07 mmol) and kiln dried powdered molecular sieves (1.0 g) in THF (20 ml) at room temperature dimensionful (1,49 g, and 5.30 mmol), and then immediately N-iodosuccinimide (2.65 g, 1.18 mmol). Analysis by thin-layer chromatography (hexane:ethyl acetate, 1:1) reaction mixture after 2.5 hours showed that the reaction was completed by approximately 60%. After this was added N-iodosuccinimide (175 mg, 0.78 mmol) and the reaction mixture was stirred for another 30 minutes, after which analysis by thin-layer chromatography showed that the reaction is completed in a separating funnel, containing 100 ml of saturated sodium bicarbonate solution and 20 ml of 5% aqueous sodium thiosulfate solution. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated in vacuum. The residual oil was purified by flash chromatography (hexane:ethyl acetate, gradient elution from 3:1 to 1:1) and received 750 mg (62% yield of the target specified in the title compound as a white solid.

1H NMR (CDCl3, 360 MHz): 8,10 (2H, m), 7,79 (2H, m), 7,65 - 7,24 (26H, m), 7,10 (1H, m), 6,41 (1H, s), of 6.20 (1H, m), 5,79 (1H, DD, J = 8.8 and 3.6 Hz), the 5.65 (1H, d, J = 7,0 Hz), 5,52 (1H, m), 5,20 (1H, d, J = 11.8 Hz), 5,11 (1H, d, J = 11.8 Hz), 5,04 - 4,85 (6H, m), 4.75 V - 4,60 (4H, m), 4,30 (1H, d, J = 8,4 Hz), 4,15 (1H, d, J = 8,4 Hz), 3,92 (1H, d, J = 7,0 Hz), 2,65 - 11,0 (22H, incl. the singlets at 2,48, 2,19, 1,95, 1,80, 1,20, 1,10, 3H each).

(f) receiving triethanolamine salt 2'-O-phosphonomethylglycine

To a solution of 2'-O-(dibenzylideneacetone)-7-O - benzyloxycarbonylglycine (500 mg, 0,382 mmol) in ethyl acetate (40 ml) in a Parr flask was added palladium (10%) on coal. The flask was attached to a Parr apparatus, and the reaction mixture was subjected to hydrogen at 50 psi (3.5 kg/cm2). The reaction mixture was dissolved in 6.5 h and then filtered through a funnel set aside the solution was concentrated in vacuum. The crude solid is suspended in approximately 5.0 ml of ethyl acetate, and decantation of the solvent. This process was repeated three times, resulting in the received specified in the header triethanolamine salt (300 mg) with a purity of 87% in the determination by HPLC-analysis. Further purification of this compound by chromatography was carried out on a C18 column (water:acetonitrile, 3: 1) gave specified in the title target compound (120 mg, 34%) with a purity of 95% in the determination by HPLC.

1H NMR (CD CD3D2O, 300 MHz): 9,05 (1H, d, J = 8.7 Hz), 8,15 - 7,12 (21H, m), 6,40 (1H, m), equal to 6.05 (1H, m), 5,69 - of 5.55 (2H, m), 5,01 - 4,85 (6H, m), 4,35 (1H, m), 4,14 (2H, m), 3.96 points - of 3.85 (6H, m) of 3.25 (1H, d, J = 7,1 Hz).

1. Phosphonoacetate esters taxonomic derivatives of the formula

< / BR>
where R1- hydroxy-OCH2(OCH2)mOP(O) (OH)2or OC(O)ORx;

R2'is hydrogen and R2- hydroxy, -OCH2(OCH2)mOP(O) (OH)2or-OC(O)ORxor R2'- fluorine,

R2is hydrogen;

R3is hydrogen, hydroxy, acetoxy;

R4, R5- independent (C1- C6)alkyl, or Z - R6';

Z is a direct link;

R6'- aryl or heteroaryl,

Rx- (C1- C6)alkyl;

p = 0 or 1;

R6and R7>mOP(O) (OH)2;

m = 0 or 1,

or their pharmaceutically acceptable salts.

2. Connection on p. 1, where R2'is hydrogen and R2- OCH2OP(O)(OH)2or its pharmaceutically acceptable salt.

3. Connection on p. 2, where R1- hydroxy - or OC(O)ORxand Rx- (C1- C6)alkyl.

4. Connection on p. 3, where R3is hydrogen, hydroxy or acetoxy.

5. Connection on p. 3, where R4(O)pis phenyl or tributoxy.

6. Connection on p. 3, where R5- phenyl, 2-furyl or 2-thienyl.

7. Connection on p. 1, which represents a 2'-0-(etoxycarbonyl)-7-0-(phosphonomethyl)-paclitaxel or its pharmaceutically acceptable salt.

8. Connection on p. 7, which represents its sodium salt.

9. Connection on p. 7, which represents his triethanolamine salt.

10. Connection on p. 7, which represents his triethylamine salt.

11. Connection on p. 7, which represents its arginine salt.

12. Connection on p. 7, which represents a lysine salt.

13. Connection on p. 7, which represents his ethanolamine salt.

16. Connection on p. 15, which represents its sodium salt.

17. Connection on p. 1, which is 3'-N-desbenzoyl-3'-N-(tert-butyloxycarbonyl)-3'-(2-furyl)-2'-0-ethoxycarbonyl-7-0-phosphonomethylglycine or its pharmaceutically acceptable salt.

18. Connection on p. 17, which represents his triethanolamine salt.

19. Connection on p. 1, which is 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2-thienyl)-2'-0-ethoxycarbonyl-7-0-phosphonomethylglycine or its pharmaceutically acceptable salt.

20. Connection on p. 19, which represents his triethanolamine salt.

21. Connection on p. 1, where R1-OCH2OP(O) (OH)2or its pharmaceutically acceptable salt.

22. Connection on p. 21, where R2'is hydrogen, R2is hydrogen, hydroxy or-OC(O)ORxand Rx- (C1- C6)alkyl.

23. Connection on p. 22, where R3is hydrogen, hydroxy or acetoxy.

24. Connection on p. 22, where R4(O)pis phenyl or tributoxy.

25. Soemer)paclitaxel or its pharmaceutically acceptable salt.

27. Connection on p. 1, where R1and R2both OCH2OP(O)(OH)2or its pharmaceutically acceptable salt.

28. Connection on p. 1, which is 2', 7-0-bis(phosphonomethyl)paclitaxel or its pharmaceutically acceptable salt.

29. Connection on p. 28, which represents its sodium salt.

30. Connection on p. 1, where R1-OCH2OCH2OP(O)(OH)2or its pharmaceutically acceptable salt.

31. Connection on p. 1, which represents a 2'-0-phosphonomethylglycine or its pharmaceutically acceptable salt.

32. Connection on p. 31, which represents his triethanolamine salt.

33. Connection on p. 1, which is a 10-deacetyl-3'-N-desbenzoyl-3'-N-(tert-butyloxycarbonyl)-10-0-(phosphonomethyl)paclitaxel or its pharmaceutically acceptable salt.

34. Connection on p. 33, which represents his triethanolamine salt.

35. 7-0-methylthiopyrimidin III formula

< / BR>
or C13the metal alkoxide.

36. Phosphonoacetate esters taxonomic derivatives of the formula

< / BR>
where R1bis hydroxy, OCH2SCH3or OC(O)ORx or R2'- fluorine, and R2bis hydrogen;

R3bis hydroxy, protected trialkylsilyl acetoxy group;

R4, R5- independent (C1- C6)alkyl, or Z - R6';

R6'- aryl or heteroaryl;

Rx- (C1- C6)alkyl,

p = 0 or 1;

R6band R7btogether form oxoprop provided that at least one of R1b, R2b- OCH2SCH3.

37. Connection on p. 36, which is 7-0-methyldiethylamine.

38. Connection on p. 36, which represents a 2'-0-(benzyloxycarbonyl)-7-0-methyldiethylamine.

39. Connection on p. 36, which represents a 2'-0-(etoxycarbonyl)- 7-0-methyldiethylamine.

40. Connection on p. 36, which represents a 2'-0-(methylthiomethyl)-7-0-(triethylsilyl)-paclitaxel.

41. Connection on p. 36, which represents 2-0-(methylthiomethyl)paclitaxel.

42. Connection on p. 36, which represents a 2',7-0-bis(methylthiomethyl)paclitaxel.

43. Connection on p. 36, which represents a 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2-furyl)-7-0-methyldiethylamine.

44. Connect dicarbonyl-7-0-methyldiethylamine.

45. Connection on p. 36, which represents a 3'-N-desbenzoyl-3'-destiny-3'-N)(tert-butyloxycarbonyl)-3'-(2-thienyl)-7-0-methyldiethylamine.

46. Connection on p. 36, which represents a 3'-N-desbenzoyl-3'-destiny-3'-N-(tert-butyloxycarbonyl)-3'-(2-thienyl)-2'-0-ethoxycarbonyl-7-0-methyldiethylamine.

47. The connection formulas

< / BR>
where R2', R2b, R3b, R4, R5, R6b, R7band p are defined above.

48. Connection on p. 47, which represents a 2'-0-(methylthiomethyl)-7-0-criticalimportance.

49. Connection on p. 47, which represents a 2'-0-(methylthiomethyl)-7-0-benzyloxycarbonylglycine.

50. Phosphonoacetate esters taxonomic derivatives of the formula

< / BR>
where R1k- hydroxy, -OCH2OP(O)(OCH2Ry)2-or-OC(O)ORx;

R2'is hydrogen, R2chydroxy, hydroxy, protected trialkylsilyl group, OCH2OP(O)(OCH2Ry)2or R2'- fluorine,

R2cis hydrogen; R3c- acetoxy; Rx- phosphonoamidate group;

R6cand R7ctogether form oxoprop provided that at least one of R1c, R2c6
;

z is a direct link;

R6'- aryl or heteroaryl;

Rx- (C1- C6)alkyl,

p = 0 or 1;

Ry- phosphonoamidate group.

51. The connection formulas

< / BR>
where R2', R2c, R3c, R4, R5, R6c, R7c, Ryand p are defined above.

52. Antitumor pharmaceutical composition containing the active principle, presents derived taxane, and pharmaceutically acceptable carriers, wherein as derived taxane contains an effective antitumor amount of a compound under item 1.

53. Method of inhibiting growth of a tumor in a mammal, comprising the introduction into the organism of a mammal antineoplastic agents, characterized in that as antitumor agents is administered an effective amount of the compounds under item 1.

54. The method according to p. 50, characterized in that it includes oral introduction into the body of antineoplastic agents.

Priority points:

24.12.92 - PP.1-17, 22-31, 34, 37-39, 47;

17.08.93 remaining claims.

 

Same patents:

, 20-epoxy-1,2, 4,7; , 1013th- hexahydroxy-11-en-9-it is complex ester group in position 13 formed (2r, 2s)-n-hexanoyl-3 - phenylisoxazol, method thereof and pharmaceutical composition" target="_blank">

The invention relates to a new taxane, namely 4,10-diacetate 2-benzoate 5, 20-epoxy-1,2, 4,7, 1013th-hexahydroxy-11-EN-9-it is complex ester group in position 13 formed (2R, 2S)-N-hexanoyl-3-phenylisoxazol having the structural formula I, which are extracted from the bark of the roots of plants of the genus Taxus extraction

The invention relates to a new method of obtaining 7-trialkylsilyl-baccatin III General formula 1

< / BR>
in which the symbols R are identical or different, denote alkyl radicals with 1-4 carbon atoms, if appropriate substituted phenyl radical, from 10-desacetyl-baccatin III formula II

< / BR>
In the General formula (1) each of the symbols R represents preferably a linear or branched alkyl radical with 1-4 carbon atoms

The invention relates to antiparasitics agents and, in particular, to compounds related to the avermectins and milbemycin, but containing substituents at the 3-position

The invention relates to the field of medicine and for the pharmaceutical solution of antitumor action of its receipt and perfusion solution

The invention relates to pharmaceutical industry and relates to antitumor compositions containing the derivatives taxane

, 20-epoxy-1,2, 4,7; , 1013th- hexahydroxy-11-en-9-it is complex ester group in position 13 formed (2r, 2s)-n-hexanoyl-3 - phenylisoxazol, method thereof and pharmaceutical composition" target="_blank">

The invention relates to a new taxane, namely 4,10-diacetate 2-benzoate 5, 20-epoxy-1,2, 4,7, 1013th-hexahydroxy-11-EN-9-it is complex ester group in position 13 formed (2R, 2S)-N-hexanoyl-3-phenylisoxazol having the structural formula I, which are extracted from the bark of the roots of plants of the genus Taxus extraction

The invention relates to the field of macrolides

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of borrelidin of the general formula (I)

wherein R represents the group of the general formulae -COOR1, -CONR2R3, -CONR4CONR2R5 or -CH2OR6 wherein R1 represents (C2-C6)-alkyl group, (C1-C6)-alkyl group substituted with hydroxyl group or 5-8-membered saturated nitrogen-containing heterocyclic group (it can comprise oxygen atom in addition to nitrogen atom) or 5-6-membered nitrogen-containing aromatic heterocyclic group or (C3-C6)-cycloalkyl group; R2 and R3 are similar or different and represent independently hydrogen atom or (C1-C6)-alkyl group that can be substituted optionally with hydroxyl, (C2-C5)-alkoxycarbonyl or 5-8-membered saturated nitrogen-containing heterocyclic group (it can comprises oxygen atom in addition to nitrogen atom) or 5-6-membered aromatic homocyclic group or aromatic heterocyclic group comprising oxygen and/or nitrogen atom, 5-6-membered cycloalkyl or heteroaryl group; R4 and R5 are similar or different and represent independently hydrogen atom or (C3-C6)-cycloalkyl group; R6 represents hydrogen atom; also, invention relates to tautomers, solvates of these compounds, their mixtures and acid-additive salts. Also, invention relates to pharmaceutical compositions comprising compounds of the general formula (I) as an active component. Angiogenesis inhibitors of the present invention inhibit formation of new vessels in tissues of live organisms and can be used for prophylaxis and inhibition of the angiogenesis process arising in the tumor proliferation, and for prophylaxis of formation of tumor metastasis. Invention provides preparing new derivatives of borrelidin eliciting the value physiological effect.

EFFECT: valuable medicinal properties of compounds.

8 cl, 15 ex

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