RU2136673C1 - Taxane derivative phosphonooxymethyl esters, intermediate compounds, antitumor pharmaceutical composition, method of tumor growth suppression in mammals - Google Patents

Abstract

FIELD: organic chemistry, pharmacy. SUBSTANCE: invention relates to taxane derivative phosphonooxymethyl esters of the formula (I)

where R¹ - hydroxy-group, -OCH₂(OCH₂)_mOP(O)(OH)₂ or OC(O)OR^x; R^2′ - hydrogen atom; R² - hydroxy-group, -OCH₂(OCH₂)_mO-P(O)(OH)_2′-OC(O)OR^x; or R^2′ - fluorine atom and R² - hydrogen atom; R³ - H, hydroxy-, acetoxy-group; R⁴,R⁵ is C_1-6-alkyl or Z-R⁶ where Z - a direct bond; R^6′ - aryl or heteroaryl; R^x is C_1-6-alkyl; p = 0 or 1; R⁶,R⁷ in common form oxo-group at condition that at least one of R¹,R² is OCH₂(OCH₂)_mOP(O)(OH)₂; m = 0 or 1, or their pharmaceutically acceptable salts that show antitumor activity. EFFECT: improved method of synthesis, antitumor activity of compounds. 54 cl, 5 tbl, 11 ex

Description

 This application is a partial continuation of the simultaneously considered application 08/108015 of these inventors, filed August 17, 1993, which in turn is a partial continuation of the application in the US N 07/996455, filed December 24, 1992 / now canceled /. US Application N 08/108015 is incorporated herein by reference in its entirety.

 The present invention relates to antitumor compounds. In particular, in accordance with the present invention, novel taxane derivatives, their pharmaceutical compositions and their use as antitumor substances are provided.

 Taxol (paclitaxel) is a natural product extracted from the Pacific yew bark, Taxus blevifolia. It turned out that it has excellent antitumor activity in animal models (in vivo), and recent studies have explained its unique nature of action, including abnormal turbulin polymerization and impaired mitosis. He is currently undergoing clinical trials against ovarian, breast and other types of cancer in the United States and France, and preliminary results have confirmed that he is the most promising chemotherapeutic drug. Clinical trials of paclitaxel have been reviewed by Rowinsky and Donehower, "The Clinical Pharmacology and Use Antimicrotubule Agents in Cancer Chemotherapeutics", Pharmac. Ther., 52: 35-84, 1991.

 Recently, a semisynthetic analogue of paclitaxel was also discovered under the name Taxotere (brand name), which showed good antitumor activity in animal models. Taxotere is also undergoing clinical trials in Europe and the United States of America. The structure of paclitaxel and Taxotere with the paclitaxel molecule traditional for the numbering system is shown below.

Таксол: R = Ph; R' = ацетил
Таксотер: R = трет-бутокси; R' = водород.

Taxol: R = Ph; R '= acetyl
Taxotere: R = tert-butoxy; R '= hydrogen.

One of the disadvantages of paclitaxel is its very organic solubility in water, which requires the preparation of dosage forms in non-aqueous pharmaceutical carriers. One of the commonly used carriers is Cremophor EL, which itself has undesirable side effects on humans. Therefore, a number of research groups have obtained water-soluble derivatives of paclitaxel, which are disclosed in the following reference materials:
(a) Hogwitz et al. U.S. Patent No. 4,942,184;
(b) Kingston et al. U.S. Patent No. 5,096,699;
(c) Stella et al. U.S. Patent No. 4,960,790;
(d) European Patent Application 0558959 A1, published September 8, 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 of the present invention are phosphonooxymethyl esters of taxon derivatives and pharmaceutically acceptable salts thereof. The solubility of salts in water facilitates the preparation of pharmaceutical compositions.

The present invention relates to taxane derivatives having the formula (A)
T - [OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ] _n , (A)
where T is a taxane fragment bearing on the C13 carbon atom a substituted 3-amino-2-hydroxypropanoyloxy group; n is 1, 2 or 3; m = 0 or an integer from 1 to 6 inclusive; or their pharmaceutically acceptable salts.

In accordance with the present invention also provides taxane derivatives having the formula (B):
T '- [OCH ₂ (OCH ₂ ) _m SCH ₃ ] _n , (B)
where T 'is T in which non-reactive hydroxy groups are protected, m and n are those defined for formula (A).

Further, in accordance with the present invention, intermediates having the formula (C) are provided:
T '- [OCH ₂ (OCH ₂ ) _m OP (O) (OR ^y ) ₂ ] _n , (C)
where T ', m and n are such as those defined for formula (A), and R ^y is a phosphono protecting group.

In addition, in accordance with the present invention, there are provided compounds of formula (D):
13-OH-txn - [OCH ₂ (OCH ₂ ) _m SCH ₃ ] _n , (D)
where m and n are such as those defined above; and txn is a taxane fragment; or their C13 metal alkoxide.

 Still in accordance with the present invention, there is provided a method of inhibiting tumor growth in a mammalian host, comprising administering to the body of said mammalian host an effective anti-tumor amount of a compound of formula (A).

 And in accordance with the present invention, there is provided a pharmaceutical composition which comprises an anti-tumor amount of a compound of formula (A) and a pharmaceutically acceptable carrier.

 Unless otherwise specifically indicated in the description or context, the following definitions should be kept in mind. “Alkyl” means a straight or branched saturated carbon chain having from one to six carbon atoms; examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl and n-hexyl. "Alkenyl" means a straight or branched carbon chain having at least one carbon-carbon double bond and having from two to six carbon atoms; examples include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, pentenyl and hexenyl. "Alkynyl" means an unbranched or branched carbon chain having at least one triple carbon-carbon bond and from two to six carbon atoms; examples include ethynyl, propynyl, butynyl and hexynyl.

"Aryl" means an aromatic hydrocarbon having from six to ten carbon atoms; examples include phenyl and naphthyl. "Substituted aryl" means aryl substituted by at least one group selected from C _1-6 alkanoyloxy, hydroxy, halogen, C _1-6 alkyl, trifluoromethyl, C _1-6 alkoxy, aryl, C _2-6 alkenyl, C _{1 -6} alkanoyl, nitro, amino and amido. "Halogen" means fluoro, chloro, bromo and iodo.

“Phosphono” means the group —P (O) (OH) ₂ , and “phosphonooxymethoxy” or “phosphonooxymethyl ether” means the trivial group —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ . “(Methylthio) thiocarbonyl” means the group —C (S) SCH ₃ . "Methylthiomethyl" (also referred to as the abbreviation MTM) is the trivial name for the group —CH ₂ SCH ₃ .

 "Taxane fragment" (also referred to as the abbreviation txn) means fragments containing a taxane skeleton of twenty carbon atoms represented by the structural formula shown below in absolute configuration.

Показанная выше система нумерации, использованная в данном описании, является традиционной для номенклатуры таксанов, например, обозначение C1 относится к углеродному атому, отмеченному цифрой "1"; C5-C20 оксетан относится к оксетановому кольцу, снабженному y углеродных атомов 4, 5 и 20 атомом кислорода; и C₉ окси к атому кислорода, присоединенному к углеродному атому, отмеченному цифрой "9", причем упомянутый атом кислорода может быть представлен оксогруппой, α- или β- гидрокси или α- или β- ацилокси.

The numbering system shown above, used in this description, is traditional for the nomenclature of taxanes, for example, the designation C1 refers to the carbon atom marked with the number "1"; C5-C20 oxetane refers to an oxetane ring provided with y carbon atoms 4, 5 and 20 of an oxygen atom; and C ₉ oxy to an oxygen atom attached to the carbon atom marked with the number “9”, said oxygen atom being represented by an oxo group, α or β hydroxy, or α or β acyloxy.

(X - неводородная группа и X' - водород или неводородная группа). Стереохимия этого остатка такая же, как у боковой цепи паклитаксела. Эта группа иногда названа в данном описании как "боковая цепь у C13"."Substituted 3-amino-2-hydroxypropanoyloxy" means a residue represented by the formula

(X is a non-hydrogen group and X 'is hydrogen or a non-hydrogen group). The stereochemistry of this residue is the same as that of paclitaxel side chain. This group is sometimes referred to herein as “C13 side chain.”

“Taxane derivative” (abbreviated T) refers to a compound having a taxane moiety carrying a side chain at C ₁₃ .

 “Heteroaryl” means a five- or six-membered aromatic ring containing at least one to four non-carbon atoms selected from oxygen, sulfur and nitrogen. Examples of heteroaryl include thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl, pyrazinyl.

 "Phosphonic protecting groups" means fragments that can be used to block (protect) the phosphonic functional group; preferably, such protecting groups are those that can be removed by methods that do not significantly affect the rest of the molecule. Suitable phosphonooxy protecting groups are those well known to those skilled in the art, including, for example, benzyl and allyl groups.

 “Hydroxy protecting groups” include, but are not limited to, ethers such as methyl, tert-butyl, benzyl, para-methoxybenzyl, para-nitrobenzyl, allyl, trityl, methoxymethyl, methoxyethoxymethyl, ethoxyethyl, tetrahydropyranyl and tetrahydryl ether, tetrahydryl, tetrahydryl, tetrahydryl, as trimethylsilyl and tert-butyldimethylsilyl ethers; esters such as benzoyl, acetyl, phenoacetyl, formyl, mono-, di- and trihaloacetyl, such as chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl; and carbonates such as methyl, ethyl, 2,2,2-trichloroethyl, allyl, benzyl and paranitrophenyl carbonates.

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

 In these textbooks, you can also find methods for introducing and removing protective groups.

 "Pharmaceutically acceptable salt" means a metal or amine salt of an acid phosphono group in which the cation does not contribute significantly to the toxicity or biological activity of the active compound. Suitable metal salts include lithium, sodium, potassium, calcium, barium, magnesium, zinc and aluminum salts. Preferred metal salts are sodium and potassium salts. Suitable amine salts are, for example, ammonia, tromethamine (TRIS), triethylamine, procaine, benzathine, dibenzylamine, chlorprocaine, choline, diethanolamine, triethanolamine, ethylenediamine, glucamine, N-methylglucamine, lysine, arginine, and many other ethanol salts. Preferred amine salts are lysine, arginine and N-methylglucamine salts.

In the description and claims, -OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ encompasses both free acid and its pharmaceutically acceptable salts, unless the meaning specifically indicates that free acid is meant.

In accordance with the present invention, taxane derivatives of the formula (A) are provided
T --- [OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ] _n , (A)
where T is a taxane moiety carrying a substituted 3-amino-2-hydroxypropanoyloxy group on a C13 carbon atom; n is 1, 2 or 3; m = 0 or an integer from 1 to 6, inclusive,
or their pharmaceutically acceptable salts.

 In one embodiment, the taxane moiety contains at least the following functional groups: C1 — hydroxy, C2 — benzyloxy, C4 — acetyloxy, C5 — C20 oxetane, C9 — oxy, and C11 — C12 double bond.

где R^2e' - водород и R^2e - водород, гидрокси, -OC(O)R^x или -OC(O)OR^x; или R^2e - водород и R^2e' - фтор; R^3e - водород, гидрокси, -OC(O)R^x, C_1-6 алкилокси или -OC(O)OR^x; один из R^6e и R^7e - водород, а другой - гидрокси или -OC(O)OR^x; R^6e и R^7e вместе образуют оксогруппу; R^x - такой, как тот, что определен ниже.In a preferred embodiment, the taxane moiety is derived from a residue having the formula

where R ^{2e '} is hydrogen and R ^2e is hydrogen, hydroxy, -OC (O) R ^x or -OC (O) OR ^x ; or R ^2e is hydrogen and R ^{2e '} is fluoro; R ^3e is hydrogen, hydroxy, —OC (O) R ^x , C _1-6 alkyloxy or —OC (O) OR ^x ; one of R ^6e and R ^7e is hydrogen, and the other is hydroxy or —OC (O) OR ^x ; R ^6e and R ^7e together form an oxo group; R ^x - such as that defined below.

где R^1e - водород или -C(O)R^x, -C(O)OR^x; R⁴ и R⁵ - независимо C_1-6 алкил, C_2-6 алкенил, C_2-6 алкинил или -Z-R⁶;
Z - прямая связь, C_1-6 алкил или C_2-6 алкенил; R⁶ - арил, замещенный арил, C_3-6 циклоалкил или гетероарил; и R^x - C_1-6 алкил, необязательно замещенный на один - шесть одинаковых или разных атомов галогена, C_3-6 циклоалкил, C_2-6 алкенил или радикал формулы

где D - связь или C_1-6 алкил; и R^a, R^b и R^c - независимо водород, амино, C_1-6 алкиламино, ди-C_1-6 алкиламино, галоген, C_1-6 алкил или C_1-6 алкокси; p = 0 или 1.In another embodiment, the side chain at C13 is derived from a residue having the formula

where R ^1e is hydrogen or —C (O) R ^x , —C (O) OR ^x ; R ⁴ and R ⁵ are independently C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl or —ZR ⁶ ;
Z is a direct bond, C _1-6 alkyl or C _2-6 alkenyl; R ⁶ is aryl, substituted aryl, C _3-6 cycloalkyl or heteroaryl; and R ^x is C _1-6 alkyl optionally substituted with one to six identical or different halogen atoms, C _3-6 cycloalkyl, C _2-6 alkenyl or a radical of the formula

where D is a bond or C _1-6 alkyl; and R ^a , R ^b and R ^c are independently hydrogen, amino, C _1-6 alkylamino, di-C _1-6 alkylamino, halogen, C _1-6 alkyl or C _1-6 alkoxy; p = 0 or 1.

In a preferred embodiment, R ⁴ is C _1-6 alkyl and p = 1 or R ⁴ is -ZR ⁶ and p = 0. More preferably R ⁴ (O) _p is tert-butoxy, phenyl, isopropyloxy, n-propyloxy or n- butoxy.

In another preferred embodiment, R ⁵ is C _2-6 alkenyl or —ZR ⁶ and Z and R ⁶ are as defined above. More preferably, R ⁵ is phenyl, 2-furyl, 2-thienyl, isobutenyl, 2-propenyl or C _3-6 cycloalkyl.

где R¹ - гидрокси, -OCH₂(OCH₂)_mОP(O)(OH)₂, - OC(O)R^x или - OC(O)OR^x; R^2' - водород и R² - водород, гидрокси, -ОСH₂(OCH₂)_mOP(O)(OH)₂ или -OC(O)OR^x; или R^2' - фтор, и R² - водород; R³ - водород, гидрокси, ацетокси, -OCH₂(OCH₂)_mOP(O)(OH)₂ или -OC(O)OR^x; один из R⁶ или R⁷ - водород, а другой - гидрокси, C_1-6 алканоилокси или -OCH₂(OCH₂)_mOP(O)(OH)₂; или R⁶ и R⁷ вместе образуют оксогруппу; при условии, что, по крайней мере, R¹, R², R³, R⁶ или R⁷ - -OCH₂(OCH₂)_mOP(O)(OH)₂, R⁴, R⁵, R^x, m и p - такие, как те, что представлены выше; или его фармацевтически приемлемыми солями.In another embodiment, the compound of formula (A) may be more specifically represented by formula (I)

where R ¹ is hydroxy, -OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ , - OC (O) R ^x or - OC (O) OR ^x ; R ^{2 '} is hydrogen and R ² is hydrogen, hydroxy, -OSH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or -OC (O) OR ^x ; or R ^{2 ′} is fluoro and R ² is hydrogen; R ³ is hydrogen, hydroxy, acetoxy, —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or —OC (O) OR ^x ; one of R ⁶ or R ⁷ is hydrogen and the other is hydroxy, C _1-6 alkanoyloxy or —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ; or R ⁶ and R ⁷ together form an oxo group; with the proviso that at least R ¹ , R ² , R ³ , R ⁶ or R ⁷ is —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ , R ⁴ , R ⁵ , R ^x , m and p are such as those presented above; or its pharmaceutically acceptable salts.

In the compounds of formula (I), examples of R ^x include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, chloromethyl, 2,2,2-trichloroethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, ethenyl, 2-propenyl, phenyl, benzyl, bromophenyl, 4-aminophenyl, 4-methylaminophenyl, 4-methylphenyl, 4-methoxyphenyl, etc. Examples of R ⁴ and R ⁵ include 2-propenyl, isobutenyl, 3-furanyl (3-furyl), 3- thienyl, phenyl, naphthyl, 4-hydroxyphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-trifluoromethylphenyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, ethenyl, 2-propenyl, 2- propynyl, benzyl, phenethyl, feni lathenyl, 3,4-dimethoxyphenyl, 2-furanyl (2-furyl), 2-thienyl, 2- (2-furanyl) ethenyl, 2-methylpropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexylmethyl, cyclohexylethyl, etc.

In a preferred embodiment, the present invention provides compounds of formula (I) wherein R ⁵ is C _2-6 alkenyl or —ZR ⁶ , and Z and R ⁶ are preferably as defined above. More preferably, R ⁵ is phenyl, 3-furyl, 3-thienyl, 2-propenyl, isobutenyl, 2-furyl, 2-thienyl or C _3-6 cycloalkyl.

In another preferred embodiment, in the compounds of formula (I), R ⁴ is C _1-6 alkyl, with p = 1; or R ⁴ is —ZR ⁶ and Z and R ⁶ are as defined above, with p = 0. More preferably R ⁴ (O) _p is tert-butoxy, phenyl, isopropyloxy, n-propyloxy, n-butoxy.

In another preferred embodiment, the present invention provides compounds of formula (I) wherein R ¹ is —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ . In a more preferred embodiment, R ² is hydroxy, —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or —OC (O) R ^x , and R ^x is preferably C _1-6 alkyl. In another preferred embodiment, R ³ is hydroxy or acetoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), wherein R ² is —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ; R ¹ is hydroxy or —OC (O) R ^x ; and R ³ is hydrogen, hydroxy, acetoxy, —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or —OC (O) OR ^x ; and R ^x is as defined above. In a more preferred embodiment, R ¹ is hydroxy or —OC (O) OR ^x and R ^x is preferably C _1-6 alkyl; and R ³ is hydroxy or acetoxy.

In another preferred embodiment, the present invention provides a compound of formula (I) in which R ³ is —OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ; R ¹ is hydroxy or —OC (O) OR ^x ; R ^{2 '} is hydrogen, and R ² is hydrogen, hydroxy or —OC (O) OR ^x ; or R ^{2 '} is fluoro and R ² is hydrogen; and R ^x is as defined above. In a more preferred embodiment, R ¹ is hydroxy or —OC (O) OR ^x and R ^x is preferably C _1-6 alkyl. In another more preferred embodiment, R ² is hydroxy.

 In another preferred embodiment, m = 0 or 1 when a phosphonooxymethoxy group is present on the C7 taxane moiety.

 Preferred pharmaceutically acceptable salts of the compounds of formula (A) are alkali metal salts including lithium, sodium and potassium salts, and amine salts including salts of triethylamine, triethanolamine, ethanolamine, arginine, lysine and N-methylglucamine. Even more preferred salts are salts of arginine, lysine and N-methylglucamine.

 The most preferred taxane derivatives of formula (A) include the following compounds: (1) 7-O-phosphonooxymethylpaclitaxel; (2) 2'-O- (ethyloxycarbonyl) -7-O-phosphonooxymethylpaclitaxel; (3) 2'-O-phosphonooxymethylpaclitaxel; (4) 2 ', 7-bis-O- (phosphonooxymethyl) paclitaxel; (5) 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) - 3 '- (2-furyl) -2'-O-ethyloxycarbonyl-7-O-phosphonoxymethylpaclitaxel; (6) 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) - 3 '- (2-thienyl) -2'-O-ethyloxycarbonyl-7-O-phosphonooxymethylpaclitaxel; (7) 10-deacetyl-3'-N-desbenzoyl-3'-N- (tert-butyloxycarbonyl) - 10-O- (phosphonooxymethyl) paclitaxel; (8) 2'-O-phosphonooxymethoxymethylpaclitaxel and their corresponding pharmaceutically acceptable salts, in particular salts of sodium, potassium, arginine, lysine, N-methylglucamine, ethanolamine, triethylamine and triethanolamine.

Compounds of formula (A) can be prepared from the starting material — taxane derivatives T - [OH] _n , where T and n are as defined above. The nature of T- [OH] _{n is} not particularly limited as long as there is at least one reactive hydroxy group present on the taxane moiety or side chain of C13 to allow for the formation of an ether bond (phosphonoxymethyl ether). It is understood that a reactive hydroxy group can be directly attached to the propanoyloxy backbone of C13 (e.g., the 2'-hydroxy group of paclitaxel) or to the taxane skeleton (e.g. the 7-hydroxy group of paclitaxel), or it may be present on the substituent on the side chain at C13 or on Deputy on the taxane skeleton. To obtain the compounds of formula (A), the reaction sequence shown in Scheme I can be used.

В схеме I T' - таксановое производное, в котором не вступающие в реакцию гидроксигруппы уже заблокированы (защищены); R^y - фосфонозащитная группа; n и m - такие, как определенные выше. Следовательно, соответственно защищенное T', имеющее одну или несколько реакционноспособных гидроксигрупп, сначала превращают в соответствующий метилтиометиловый эфир формулы (B). При использовании в качестве примера паклитаксела T' может представлять собой сам паклитаксел (для осуществления 2', 7-бисметилтиометилирования), 7-O-триэтилсилилпаклитаксел или 2'-O-этоксикарбонилпаклитаксел. Соединение формулы (B), где m = 0, может быть получено путем обработки T'-[OH]_n диметилсульфоксидом с уксусным ангидридом или диметилсульфидом и органическим пероксидом. Указанные реакции описаны более полно ниже.

In the IT 'scheme, a taxane derivative in which non-reactive hydroxy groups are already blocked (protected); R ^y is a phosphonoprotective group; n and m are as defined above. Therefore, a suitably protected T ′ having one or more reactive hydroxy groups is first converted to the corresponding methylthiomethyl ester of formula (B). When used as an example of paclitaxel, T 'may be paclitaxel itself (for 2', 7-bismethylthiomethylation), 7-O-triethylsilylpaclitaxel or 2'-O-ethoxycarbonylpaclitaxel. The compound of formula (B), where m = 0, can be obtained by treating T ′ - [OH] _n with dimethyl sulfoxide with acetic anhydride or dimethyl sulfide and organic peroxide. These reactions are described more fully below.

 MTM ether having one intermediary (intermediate) methyleneoxy unit (i.e., compounds of formula (B), where m = 1) can be obtained in several possible ways. In this method, a compound of formula (B), where m = 0, is reacted with N-iodosuccinimide (NIS) and methylthiomethanol to continue the chain to one methyleneoxy unit.

T' -- [OCH₂OCH₂SCH₃]_n.T '- [OCH ₂ SCH ₃ ] _n + nCH ₃ SCH ₂ --OH

T 'is [OCH ₂ OCH ₂ SCH ₃ ] _n .

 The methylthiomethanol compound and its preparation are reported to Syn. Comm., 1986, 16 (13): 1607-1610.

 In an alternative method, T-alkoxide (Ad) obtained by treating a compound of formula (Aa) with a base such as n-butyllithium, lithium diisopropylamide or lithium hexamethyldisilazide is reacted with chloromethylmethylthiomethyl ether to obtain a compound of formula (B), where m = 1.

Соединение (Ae) получают введением метилтиометоксида (полученного из метилтиометанола путем обработки основанием, таким как н-бутиллитий, диизопропиламид лития или гексаметилдисилазид лития) во взаимодействие с хлориодметаном. Соединение (Ae) может быть также получено путем обработки 1,1'-дихлордиметилэфира (ClCH₂OCH₂Cl) стехиометрическим количеством или менее (например, около 0,8 эквивалента) иодида натрия, а затем тиометоксида натрия. О 1,1'-дихлордиметиловом эфире сообщено в Ind. J. Chem., 1989, 28B, стр. 454 - 456.

Compound (Ae) is prepared by introducing methylthiomethoxide (obtained from methylthiomethanol by treatment with a base such as n-butyllithium, lithium diisopropylamide or lithium hexamethyldisilazide) into reaction with chlorodiomethane. Compound (Ae) can also be obtained by treating 1,1'-dichlorodimethylether (ClCH ₂ OCH ₂ Cl) with a stoichiometric amount of less or less (for example, about 0.8 equivalents) of sodium iodide and then sodium thiomethoxide. 1,1'-dichlorodimethyl ether is reported in Ind. J. Chem., 1989, 28B, pp. 454 - 456.

In yet another method, a compound of formula (Ae) is reacted with bis (MTM) ether, CH ₃ SCH ₂ OCH ₂ SCH ₃ , and NIS, and a compound of formula (B) is obtained, where m = 1.

T '- [OH] _n + nCH ₃ SCH ₂ OCH ₂ SCH ₃ --->T' - [OCH ₂ OCH ₂ SCH ₃ ] _n ,
Bis (MTM) ether is prepared by introducing 1,1'-dichlorodimethyl ether into reaction with sodium iodide, and then with sodium thiomethoxide.

 The above procedure using methylthiomethanol and NIS can be applied to any reagent having an MTM group to continue the chain to one methyleneoxy unit at a time. For example, a compound of formula (B), where m = 1, can be reacted with NIS to obtain a compound of formula (B) in which m = 2. The process can be repeated to obtain compounds of formula (B) in which m = 3, 4, 5 or 6.

In a second step shown in Scheme 1, methylthiomethyl ether is converted to the corresponding protected phosphonooxymethyl ether. This is accomplished by treating MTM ether with NIS and protected phosphate HOP (O) (OR ^y ) ₂ . In a third step, the phosphonoprotective group and the hydroxy protecting group (s) are removed to give compound (A). For example, a suitable phosphonoprotective group is benzyl, which can be removed by catalytic hydrogenolysis; hydroxy protecting groups such as trialkylsilyl can be removed by fluoride ion, trichloroethoxycarbonyl can be removed by zinc. Deprotection is 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 steps are described in more detail later in the description.

 A variant of the reaction sequence shown in Scheme I is presented in Scheme II.

В схеме II соединение формулы (Aa) вводят в химическое взаимодействие с соединением (Ca) и NIS и получают соединение формулы (C), у которого затем снимают защиту и получают в результате соединение формулы (A). Соединения формулы (Ca), в которой m = 0, могут быть получены сначала обработкой метилтиометанола основанием, таким как гексаметилдисилазид натрия, лития или калия, чтобы получить метилтиометоксид; метоксид затем вводят во взаимодействие с защищенным хлорфосфатом, таким как дибензилхлорфосфат, и получают требуемое соединение. Соединения формулы (Ca), в которой m = 1, могут быть получены обработкой CH₃SCH₂OCH₂Cl двузащищенным фосфатом (солью), например, дибензилфосфатами натрия, калия, тетра(н-бутил)аммония; или CH₃SCH₂OCH₂Cl может быть сначала превращено в соответствующее иодистое соединение с использованием иодида натрия до введения во взаимодействие с фосфатом. В соответствии с другим вариантом соединения формулы (Ca), в которой m = 1, могут быть получены путем обработки ClCH₂OCH₂Cl иодидом натрия с последующей обработкой тиометоксидом натрия, в результате чего получают CH₃SCH₂OCH₂SCH₃; это соединение затем обрабатывают NIS-ом и двузащищенным фосфатом, таким как дибензилфосфат, и получают требуемый продукт. Любой из упомянутых выше реагентов, имеющий МТМ группу, может быть продолжен на одно метиленокси-звено за раз путем введения упомянутого реагента во взаимодействие с метилтиометанолом и NIS.

In Scheme II, a compound of formula (Aa) is chemically reacted with compound (Ca) and NIS to give a compound of formula (C), which is then deprotected and the resulting compound of formula (A). Compounds of formula (Ca) in which m = 0 can be prepared by first treating methylthiomethanol with a base such as sodium, lithium or potassium hexamethyldisilazide to obtain methylthiomethoxide; the methoxide is then reacted with protected chlorophosphate, such as dibenzyl chlorophosphate, to give the desired compound. Compounds of formula (Ca) in which m = 1 can be obtained by treating CH ₃ SCH ₂ OCH ₂ Cl with a bi-protected phosphate (salt), for example, sodium, potassium, tetra (n-butyl) ammonium dibenzyl phosphates; or CH ₃ SCH ₂ OCH ₂ Cl may first be converted to the corresponding iodide compound using sodium iodide before being reacted with phosphate. In another embodiment, compounds of formula (Ca) in which m = 1 can be prepared by treating ClCH ₂ OCH ₂ Cl with sodium iodide, followed by treatment with sodium thiomethoxide, to give CH ₃ SCH ₂ OCH ₂ SCH ₃ ; this compound is then treated with NIS and a bi-protected phosphate such as dibenzyl phosphate, and the desired product is obtained. Any of the above reagents having an MTM group can be extended one methyleneoxy unit at a time by introducing the above reagent into reaction with methylthiomethanol and NIS.

 According to another method for preparing a compound of formula (A), T-alkoxide (Ad) is reacted with iodine phosphate as shown in Scheme III.

В схеме III иодфосфатное соединение получают путем введения во взаимодействие ClCH₂(OCH₂)_mCl с двузащищенным фосфатом, что дает ClCH₂(OCH₂)_mOP(O)(OR^y)₂, которое затем обрабатывают иодидом натрия, получая в результате целевой продукт.

In Scheme III, an iodophosphate compound is prepared by reacting ClCH ₂ (OCH ₂ ) _m Cl with bi-protected phosphate to give ClCH ₂ (OCH ₂ ) _m OP (O) (OR ^y ) ₂ , which is then treated with sodium iodide, resulting in target product.

 Another method suitable for preparing a subgroup of compounds of formula (A) in which at least one of the phosphonooxymethoxy groups is linked to a taxane moiety is shown in Scheme IV.

В схему IV m и n - такие, как определенные выше, X - неводородная группа, P - гидроксизащитная группа, txn - таксановый фрагмент. Соединения формулы (D) являются таксанами, имеющими 13 α- гидроксигруппу и один или несколько метилтиометиловых эфиров, связанных непосредственно или косвенно с таксановым скелетом; включены также C13-алкоксиды металлов формулы (D). Примером соединения формулы (D) является 7-O-метилтиометилбаккатин III:

Сочетание таксана (D) с азетидиноном аналогично тому, что показано ниже в схеме VI, и описанная процедура получения соединения формулы (Id) применима также к получению соединения формулы (Ba) [т.е. соединения формулы (B), в котором, по крайней мере, одна МТМ группа связана непосредственно или косвенно с таксановым фрагментом], если вместо соединения формулы (II) в схеме VI использовать соединение формулы (D). Таксан (D) предпочтительно сначала превращают в C13-алкоксид металла, такой как алкоксид натрия, калия или лития (предпочтительным является алкоксид лития). Азетидинон служит предшественником боковой цепи у C13. После реакции сочетания с таксаном удаляют гидроксизащитную группу P и, если требуется, свободная гидроксигруппа на боковой цепи может быть превращена в МТМ эфир или произведена в сложный эфир или карбонат, как здесь описано.Scheme IV

In Scheme IV, m and n are as defined above, X is a non-hydrogen group, P is a hydroxy protecting group, txn is a taxane fragment. The compounds of formula (D) are taxanes having 13 α-hydroxy groups and one or more methylthiomethyl esters linked directly or indirectly to the taxane skeleton; C13 metal alkoxides of metals of formula (D) are also included. An example of a compound of formula (D) is 7-O-methylthiomethylbaccatin III:

The combination of taxane (D) with azetidinone is similar to that shown in Scheme VI below, and the described procedure for preparing a compound of formula (Id) also applies to the preparation of a compound of formula (Ba) [i.e. a compound of formula (B) in which at least one MTM group is directly or indirectly linked to a taxane moiety] if instead of a compound of formula (II) in scheme VI, a compound of formula (D) is used. Taxane (D) is preferably first converted to a C13 metal alkoxide, such as sodium, potassium or lithium alkoxide (lithium alkoxide is preferred). Azetidinone serves as a precursor to the side chain of C13. After the coupling reaction with the taxane, the hydroxy protecting group P is removed and, if desired, the free hydroxy group on the side chain can be converted to MTM ester or ester or carbonate as described herein.

 Azetidinone can be prepared by the methods described below, which are generally known in the art. Compounds of formula (D) can be prepared in the usual manner described above in connection with the preparation of compounds of formula (B) using a suitably protected taxane. However, it is more appropriate to obtain them from a compound of formula (Ba) by cleaving the side chain at C13 using a borohydride such as sodium borohydride or tetrabutylammonium; for example, 7-O-MTM paclitaxel is treated with tetrabutylammonium borohydride to give 7-O-MTM baccatin III.

 The general process of Scheme I for preparing a compound of formula (A) is presented as an example more specifically in Scheme V, which illustrates the preparation of a compound of formula (I ') [t. e. the compounds of formula (I) in which m = 0]. The procedure used in this synthesis scheme is generally applicable to other taxane derivatives not specifically covered by formula (I). In addition, the procedure in Scheme V may be modified in accordance with the ideas contained herein by those skilled in the art to obtain taxane derivatives of formula (A) in which m = 1 or 2.

It should be understood that in Scheme V and elsewhere in the description, the term “hydroxy protecting group” may include carbonates (—OC (O) OR ^x ); therefore, when carbonate is used as the hydroxy protecting group, it is supposed to be removed at a later stage to form a free hydroxy group, otherwise the carbonate moiety will remain part of the final product.

In Scheme V (see end of description) R ^1a is hydroxy, protected hydroxy, —OC (O) R ^x or —OC (O) OR ^x ; R ^{2 '} is hydrogen and R ^2a is hydrogen, hydroxy, protected hydroxy or —OC (O) OR ^x ; or R ^{2 ′} is fluoro and R ^2a is hydrogen; R ^3a is hydrogen, hydroxy, protected hydroxy, acetoxy or —OC (O) OR ^x ; one of R ^6a or R ^7a is hydrogen, and the other is hydroxy, protected by hydroxy or C _1-6 alkanoyloxy; R ^6a and R ^7a together form an oxo group; with the proviso that at least one of R ^1a , R ^2a or R ^3a , R ^6a or R ^7a is hydroxy, R ^1b is hydroxy, protected hydroxy, —OCH ₂ SCH ₃ , —OC (O) R ^x or -OC (O) OR ^x ; R ^{2 ′} is hydrogen and R ^2b is hydrogen, hydroxy, protected hydroxy, —OCH ₂ SCH ₃ or —OC (O) R ^x ; or R ^{2 ′} is fluoro and R ^2b is hydrogen; R ^3b is hydrogen, hydroxy, protected hydroxy, acetoxy, —OCH ₂ SCH ₃ or —OC (O) OR ^x ; one of R ^6b or R ^7b is hydrogen and the other is hydroxy protected with hydroxy, C _1-6 alkanoyloxy or —OCH ₂ SCH ₃ ; or R ^6b and R ^7b together form an oxo group; with the proviso that one of the radicals R ^1b , R ^2b , R ^3b , R ^6b or R ^7b is —OCH ₂ SCH ₃ , R ^1c is hydroxy, protected hydroxy, —OCH ₂ OP (O) (OR ^y ) ₂ , -OC (O) R ^x or -OC (O) OR ^x ; R ^{2 ′} is hydrogen, and R ^2c is hydrogen, hydroxy, protected hydroxy, —OCH ₂ OP (O) (OR ^y ) ₂ or —OC (O) OR ^x ; or R ^{2 ′} is fluoro and R ^2c is hydrogen; R ^3c is hydrogen, hydroxy, protected hydroxy, acetoxy, —OCH ₂ OP (O) (OR ^y ) ₂ or —OC (O) OR ^x ; one of R ^6c or R ^7c is hydrogen, and the other is hydroxy, protected by hydroxy, C _1-6 alkanoyloxy or —OCH ₂ OP (O) (OR ^y ) ₂ ; with the proviso that at least one of R ^1c , R ^2c , R ^3c , R ^6c or R ^7c is —OCH ₂ OP (O) (OR ^y ) ₂ . R ^{1 '} is hydroxy, -OCH ₂ OP (O) (OH) _2' -OC (O) R ^x or -OC (O) OR ^x ; R ^{2 ″ ″} is hydrogen and R ^{2 ″} is hydrogen, hydroxy, —OCH ₂ OP (O) (OH) ₂ or —OC (O) OR ^x ; or R ^{2 ″} is fluorine and R ^{2 ″} is hydrogen; R ³ is hydrogen, hydroxy, acetoxy, —OCH ₂ OP (O) (OH) _2, or —OC (O) OR ^x ; one of R ^{6 ′} or R ^{7 ′} is hydrogen and the other is hydroxy, C _1-6 alkanoyloxy or —OCH ₂ P (O) (OH) ₂ ; with the proviso that at least one of R ^{1 ′} , R ^{2 ′} , R ^{3 ′} , R ^{6 ′} or R ^{7 ′} is —OCH ₂ OP (O) (OH) ₂ . R ⁴ , R ⁵ and R ^x are as defined above, and R ^y is a phosphono protecting group.

In the first step, the free hydroxy group of the compound of formula (Ia) is converted to the corresponding group, methylthiomethyl ether (—OCH ₂ SCH ₃ ). This conversion can be carried out by one of two procedures (Ia - dimethyl sulfoxide method) and (Ib - dimethyl sulfoxide method). The dimethyl sulfide method for converting alcohols to methylthiomethyl esters is reported in Medina et al., Tet. Lett., 1988, pp. 3773 - 3776 (relevant parts of this material are incorporated herein by reference). The dimethyl sulfoxide method is a well-known reaction, commonly called the Pammerer reaction.

 It should be noted that the reactivity of the hydroxy group differs depending on its location on the starting material, the taxane derivative of formula (Ia). Although in general the 2'-hydroxy group is more reactive in acylation reactions than the 7-hydroxy group, which in turn is more reactive than the 10-hydroxy group, it was unexpectedly found that 7-hydroxy is more easily converted to methylthiomethyl ether than the 2'-hydroxy group , The tertiary hydroxy group at C-1 is usually the least reactive. The difference in the reactivity of the hydroxy group can be used to control the location and degree of methylthiomethylation.

Thus, in the case of the compound of formula (Ia), where R ^1a and R ^2a are both hydroxy, the corresponding methylthiomethylation product is the corresponding 7-O-methylthiomethyl ether. In order to obtain a compound of formula (Ib), where R ^1b is methylthiomethoxy, and without converting the 7-hydroxy group (if any) to methylthiomethyl ether, the 7-hydroxy group is blocked with a conventional hydroxy protecting group such as triethylsilyl. Similarly, 10-methylthiomethyl ether can also be obtained without conversion of 7- and / or 2'-hydroxy groups (if present) when they are blocked with the same or different hydroxy protecting groups. Even if the 7-hydroxy group is preferred for methylthiomethylation, it is still desirable to protect the 2'-hydroxy group if the desired product is 7-monomethylthiomethyl ether.

 In addition, the reaction conditions can be adjusted in order to promote the formation of bis- or tris-methylthiomethylether taxane derivatives. For example, in the case of paclitaxel, increasing the reaction time or using a larger excess of methylthiomethylating reagents may result in a higher content of 2 ', 7-bis (methylthiomethyl) ether paclitaxel in the product mixture.

In the procedure (Ia) in Scheme V, a compound of formula (Ia) is treated with dimethyl sulfide and an organic peroxide such as benzoyl peroxide. The reaction is carried out in an inert organic solvent, such as acetonitrile, methylene chloride and the like, at a temperature favorable to the formation of the product; typically, the reaction is carried out in a temperature range of from about -40 ^° C. to ambient temperature. Dimethyl sulfide and benzoyl peroxide are used in excess with respect to the feed taxane derivative (Ia), and dimethyl sulfide is used in excess with respect to benzoyl peroxide.

 The relative amounts of raw materials used depend on the degree of methylthiomethylation to be achieved. So, when one free hydroxy group of the feed taxane derivative (Ia) is to be converted to methylthiomethyl ether, dimethyl sulfide and benzoyl peroxide can be used in up to 10-fold excess relative to the taxane derivative (Ia), and the excess of dimethyl sulfide relative to benzoyl peroxide is preferably approximately two to three times. In the case where the raw material (Ia) has both 2'- and 7-hydroxy groups, the amount of 2 ', 7-bis (methylthiomethyl) ester obtained increases with increasing relative amounts of dimethyl sulfide and benzoyl peroxide. When the target product is 2 ', 7-bis (methylthiomethyl) ether, dimethyl sulfide, it is preferable to use about 15 to 20-fold excess with respect to the feed taxane derivative, and the excess of benzoyl peroxide relative to the feed taxane derivative is about 5-10 times.

 In another embodiment, a compound of formula (Ib) can be prepared by chemically reacting a compound of formula (Ia) with dimethyl sulfoxide and acetic anhydride (Procedure Ib). This procedure is suitable for converting a non-2'-hydroxy group to its methylthiomethyl ether. In procedure (Ib), a compound of formula (Ia) is dissolved in dimethyl sulfoxide and acetic anhydride is added to the solution. The reaction is usually carried out at room temperature for 18-24 hours, and monomethylthiomethyl ether is obtained.

In a second step in the reaction sequence, methylthiomethyl ether is converted to the corresponding protected phosphonooxymethyl ether. The conversion of methylthiomethyl ether to protected phosphonooxymethyl ether can be accomplished by the general method described in Veeneman et al., Tetrahedron, 1991, v 47, pages 1547-1562 (this material is incorporated herein by reference). In this case, the compound of formula (Ib) with at least one methylthiomethyl ether group is treated with N-iodosuccinimide and protected phosphoric acid, such as dibenzyl phosphate. The reaction is carried out in an inert organic solvent, such as tetrahydrofuran or a halogenated hydrocarbon, such as 1,2-dichloroethane or methylene chloride, and, although not necessarily, in the presence of a dehydrating agent, such as molecular sieves (zeolites). A catalyst such as silver trifluoromethanesulfonate may also be added to accelerate the reaction. The reaction is carried out at a temperature in the range of from about 0 ^° C. to about room temperature, preferably at room temperature. N-iodosuccinimide and protected phosphoric acid are used in about the same molar equivalent as methylthiomethylether (Ib), but are preferably used with a slight excess, for example, about 1.3-1.5 equivalents, with respect to the compound of formula (Ib) .

 In the third stage of the reaction sequence, the phosphonoprotective group and the hydroxy protecting group (if any) are removed. Deprotection is carried out by conventional methods well known in the art, such as acid or base catalyzed hydrolysis, hydrogenolysis, reduction, and the like. For example, catalytic hydrogenolysis can be used to remove a benzyl phosphono protecting group, as well as a benzyloxycarbonyl hydroxy protecting group. Methods for removing protection can be found in ordinary textbooks, for example, Green and Wutz or Mac Omi textbooks mentioned above.

 The basic salts of the compounds of formula (I) can be formed by traditional methods, including contacting the free acid of the compounds of formula (I) with a metal base or with an amine. Suitable metal bases include 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), chlorprocaine, choline, diethanolamine, triethanolamine and the like. The basic salts can then be purified by chromatography, followed by lyophilization or crystallization.

где R₁ - -OR₆, -SR₇ или -NR₈R₉; R₂ - водород, алкил, алкенил, алкинил, арил или гетероарил; R₃ и R₄ - независимо водород, алкил, алкенил, алкинил, арил, гетероарил или ацил при условии, однако, что R₃ и R₄ не являются оба ацилом; R₅ - -COR₁₀, -COOR₁₀, -COSR₁₀, -CONR₈R₁₀, -SO₂R₁₁ или -POR₁₂R₁₃, R₆ - водород, алкил, алкенил, алкинил, арил, гетероарил, гидроксизащитная группа или функциональная группа, которая повышает растворимость в воде таксанового производного; R₇ - алкил, алкенил, алкинил, арил, гетероарил или сульфгидрилзащитная группа; R₈ - водород, алкил, алкенил, алкинил, арил, гетероарил; R₉ - аминозащитная группа; R₁₀ - алкил, алкенил, алкинил, арил, гетероарил; R₁₁ - алкил, алкенил, алкинил, арил, гетероарил, -OR₁₀ или -NR₈R₁₄; R₁₂ и R₁₃ - независимо друг от друга алкил, алкенил, алкинил, арил, гетероарил, -OR₁₀ или -NR₈R₁₄; R₁₄ - водород, алкил, алкенил, алкинил, арил, гетероарил; R₁₅ и R₁₆ - независимо друг от друга водород, гидрокси, низший алканоилокси, алкеноилокси, алкиноилокси, арилоилокси или R₁₅ и R₁₆ вместе образуют оксогруппу; R₁₇ и R₁₈ - независимо друг от друга водород, гидрокси, низший алканоилокси, алкеноилокси, алкиноилокси, арилоилокси или R₁₇ и R₁₈ вместе образуют оксогруппу; R₁₉ и R₂₀ - независимо водород или гидрокси или низший алканоилокси, алкеноилокси, алкиноилокси или арилоилокси; R₂₁ и R₂₂ - независимо водород или низший алканоилокси, алкеноилокси, алкиноилокси или арилоилокси или R₂₁ и R₂₂ вместе образуют оксогруппу; R₂₄ - водород или гидрокси или низший алканоилокси, алкеноилокси, алкиноилокси или арилоилокси; или R₂₃ и R₂₄ вместе образуют оксо или метилен или R₂₃ и R₂₄ вместе с атомом углерода, к которому они присоединены, образуют оксирановое кольцо или R₂₃ и R₂₂ вместе с углеродным атомом, к которому они присоединены, образуют оксетановое кольцо; R₂₅ - водород, гидрокси или низший алканоилокси, алкеноилокси, алкиноилокси или арилоилокси; или R₂₆ - водород, гидрокси, или низший алканоилокси, алкеноилокси, алкиноилокси или арилоилокси; или R₂₆ и R₂₅ вместе образуют оксогруппу; и R₂₇ - водород, гидрокси или низший алкокси, алканоилокси, алкеноилокси, алкиноилокси или арилоилокси; (e) таксановые производные, раскрытые в патенте США N 5227400: 3'-дезфенил-3'-(2-фурил)- или 3'-(2-тиенил)производные паклитаксела, Таксотера; (f) таксановые производные, раскрытые в EP 534709, опубликованном 31 марта 1993 г. (производные паклитаксела, в которых фенильные группы боковой цепи независимо заменены на нафтил, стирил или замещенный фенил); см. также PCT 92/09589, опубликованную 11 июня 1992 г.; (g) таксановые производные, раскрытые в EP 534707, опубликованном 31 марта 1993 г. (производные паклитаксела, в которых 3'-N-бензоильная группа замещена на этоксикарбонил или метоксикарбонил); (h) PCT заявка 93/06093, опубликованная 1 апреля 1993 г. (10-дезацетоксипроизводные паклитаксела и Таксотера); (i) EP 524093, опубликованный 20 января 1993 г. (10-, 7- или 7,10-бис-O-(N-замещенные карбамоилпроизводные таксана); (j) 9- α -гидроксианалог паклитаксела раскрыт в Klein, "Synthesis of 9-Dihydrotaxol: A New Bioactive Taxane", Tetrahedron Letters, 1993, 34 (13): 20-47-2050; (k) 14- β -гидроксианалог паклитаксела и Таксотера, полученный из 14 β -гидрокси-10-дезацетилбаккатина III, раскрыт на 205-ом национальном съезде Американского химического общества в Колорадо, 1993 г. (Med. Chem. Division, Abstract N 28); и (l) другие таксаны, такие как C7-фтортаксаны и различные C10-замещенные таксаны, раскрытые в одновременно рассматриваемой заявке 08/062687 на патент США, поданной данными заявителями 20 мая 1993 г., которая полностью включена в данное описание путем ссылки.Source Taxane Derivatives
The processes described above can be applied to any taxane derivatives of the formula T- [OH] _n to form compounds of the formula (A). Many examples of T- [OH] _n are indicated in the literature and some of them are listed below: (a) paclitaxel; (b) Taxotere (trademark); (c) 10-deacetylpaclitaxel; (d) taxane derivatives disclosed in PCT application 93/06079 (published April 1, 1993) having the formula

where R ₁ is —OR ₆ , —SR ₇ or —NR ₈ R ₉ ; R ₂ is hydrogen, alkyl, alkenyl, alkynyl, aryl or heteroaryl; R ₃ and R ₄ are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl or acyl, provided, however, that R ₃ and R ₄ are not both acyl; R ₅ - -COR ₁₀ , -COOR ₁₀ , -COSR ₁₀ , -CONR ₈ R ₁₀ , -SO ₂ R ₁₁ or -POR ₁₂ R ₁₃ , R ₆ - hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, hydroxy protecting group or a functional group that increases the solubility in water of a taxane derivative; R ₇ is an alkyl, alkenyl, alkynyl, aryl, heteroaryl or sulfhydryl protecting group; R ₈ is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl; R ₉ is an amino protecting group; R ₁₀ is alkyl, alkenyl, alkynyl, aryl, heteroaryl; R ₁₁ is alkyl, alkenyl, alkynyl, aryl, heteroaryl, —OR ₁₀ or —NR ₈ R ₁₄ ; R ₁₂ and R ₁₃ are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, —OR ₁₀ or —NR ₈ R ₁₄ ; R ₁₄ is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl; R ₁₅ and R ₁₆ are independently hydrogen, hydroxy, lower alkanoyloxy, alkenoyloxy, alkynoyloxy, aryloyloxy or R ₁₅ and R ₁₆ together form an oxo group; R ₁₇ and R ₁₈ are independently hydrogen, hydroxy, lower alkanoyloxy, alkenoyloxy, alkynoyloxy, aryloyloxy, or R ₁₇ and R ₁₈ together form an oxo group; R ₁₉ and R ₂₀ are independently hydrogen or hydroxy or lower alkanoyloxy, alkenoyloxy, alkynoyloxy or aryloyloxy; R ₂₁ and R ₂₂ are independently hydrogen or lower alkanoyloxy, alkenoyloxy, alkynyloxy or aryloyloxy, or R ₂₁ and R ₂₂ together form an oxo group; R ₂₄ is hydrogen or hydroxy or lower alkanoyloxy, alkenoyloxy, alkynyloxy or aryloyloxy; or R ₂₃ and R ₂₄ together form oxo or methylene or R ₂₃ and R ₂₄ together with the carbon atom to which they are attached form an oxirane ring or R ₂₃ and R ₂₂ together with the carbon atom to which they are attached form an oxetane ring; R ₂₅ is hydrogen, hydroxy or lower alkanoyloxy, alkenoyloxy, alkynyloxy or aryloyloxy; or R ₂₆ is hydrogen, hydroxy, or lower alkanoyloxy, alkenoyloxy, alkynyloxy or aryloyloxy; or R ₂₆ and R ₂₅ together form an oxo group; and R ₂₇ is hydrogen, hydroxy or lower alkoxy, alkanoyloxy, alkenoyloxy, alkynoyloxy or aryloyl oxy; (e) taxane derivatives disclosed in US Pat. No. 5,227,400: 3′-desphenyl-3 ′ - (2-furyl) - or 3 ′ - (2-thienyl) derivatives of paclitaxel, Taxotere; (f) taxane derivatives disclosed in EP 534709 published March 31, 1993 (paclitaxel derivatives in which the phenyl groups of the side chain are independently replaced by naphthyl, styryl or substituted phenyl); see also PCT 92/09589 published June 11, 1992; (g) taxane derivatives disclosed in EP 534707 published March 31, 1993 (paclitaxel derivatives in which the 3'-N-benzoyl group is substituted with ethoxycarbonyl or methoxycarbonyl); (h) PCT application 93/06093 published April 1, 1993 (10-deacetoxy derivatives of paclitaxel and Taxotere); (i) EP 524093 published January 20, 1993 (10-, 7- or 7,10-bis-O- (N-substituted carbamoyl derivatives of taxane); (j) 9-α-hydroxy analog of paclitaxel is disclosed in Klein, "Synthesis of 9-Dihydrotaxol: A New Bioactive Taxane ", Tetrahedron Letters, 1993, 34 (13): 20-47-2050; (k) 14-β-hydroxy analogue of paclitaxel and Taxotere derived from 14 β -hydroxy-10-deacetylbaccatin III disclosed at the 205th National Congress of the American Chemical Society in Colorado, 1993 (Med. Chem. Division, Abstract N 28); and (l) other taxanes, such as C7-fluorotaxanes and various C10-substituted taxanes, disclosed in simultaneously pending application 08/062 687 for a US patent filed by these applicants on May 20, 1993, which is incorporated herein by reference in its entirety.

The free hydroxy group (s) of the taxane derivatives can be converted by conventional methods into the corresponding esters or carbonates; for example, in compounds of formula (Ia), one of R ^1a , R ^2a or R ^3a is —OC (O) R ^x or —OC (O) OR ^x and R ^x is as defined above. Therefore, the taxane derivative T-OH can be reacted with a compound of the formula LC (O) OR ^x (L is a leaving group), such as chloroformate, in the presence of a base such as a tertiary amine to obtain the corresponding carbonate; for example, the reaction of paclitaxel with ethyl chloroformate in the presence of diisopropylethylamine gives 2'-O-ethyloxycarbonyl paclitaxel. T-OH may also be reacted with a carboxylic acid R ^x CO ₂ H or its acylating equivalent (e.g., anhydride, active ester or acyl halide) to give the corresponding ester.

In addition, taxane derivatives of T- [OH] _n can be prepared by acylating a taxane moiety having a C13 hydroxy group suitably substituted with 3-amino-2-hydroxypropanoic acid, its acylating equivalent, or its precursor. Suitable precursors for substituted 3-amino-2-hydroxypropanoic acid are, for example, azetidinones of formula (III). An example of this acylation reaction is a combination of hydroxy-protected baccatin III or hydroxy-protected 10-deacetylbaccatin III and a phenylisoserine derivative to give paclitaxel derivatives, as disclosed, for example, in Denis et al. US Pat. Nos. 4,924,011 and 4,924,012; or a combination of protected bakketin III and azetidinone giving paclitaxel and its derivatives, as disclosed in European Patent Application 400971 published December 5, 1990 (now US Pat. No. 5,175,315) and US Pat. No. 5,229,526.

A process such as that disclosed in EP 400971 (Holton process) involves reacting 1-benzoyl-3- (1-ethoxy) ethoxy-4-phenyl-2-azetidinone with 7-O-triethylsilyl baccatin III in the presence of N , N-dimethylaminopyridine and pyridine at 25 ^° C. for 12 hours, whereby paclitaxel is obtained after the removal of various hydroxy protecting groups. Ojima et al. , "New and Efficient Approaches to the Semisynthesis of Taxol and its C-13 Side Chain Analogs by Means of β-Lactam Synthon Method" Tetrahedron, 1992, 48 (34): 6985 - 7012 describes an improvement to the Holton process. The Ojima process involves first the formation of the sodium salt of 7-triethylsilylbaccatin III by sodium hydride, and then the introduction of this salt into reaction with the chiral 1-benzoyl-3- (1-ethoxy) ethoxy-4-phenyl-2-azetidinone, resulting in after removal hydroxy protecting groups receive paclitaxel. In US Pat. No. 5,229,526, Holton discloses a combination of a metal of a baccatin III alkoxide or derivative thereof with 2-azetidinone, resulting in side chain taxanes at C13. This process, as said, is highly diastereoselective, therefore, racemic mixtures of the side chain 2-azetidinone precursor can be used. As recently reported by Ojima et al., "A Highly Efficient Route to Taxotere by the β-Lactam Synthon Method", Tetrahedron Letters, 1993, 34 (26): 4149 - 4152, metal alkoxides 7,10-bis-O- (trichloroethoxycarbonyl ) -10-deacetylbaccatin III with chiral 1- (tert-butoxycarbonyl) -4-phenyl-3- (protected by hydroxy) -2-azetidinone gives Taxotere after deprotection. Related to the part described in this application, all of the above links are incorporated into this description by reference.

 The process of interaction of baccatin with azetidinone, common to obtain the compounds of formula (Ia) is shown in scheme VI (see the end of the description). Other taxane derivatives that are not specifically covered by formula (Ia) can also be prepared by this method using appropriate raw materials.

In Scheme VI, R ^{2 '} is hydrogen, and R ^2d is hydrogen protected by hydroxy or —OC (O) OR ^x ; or R ^{2 '} is fluoro and R ^2d is hydrogen; R ^3d is hydrogen, acetoxy, protected hydroxy or —OC (O) OR ^x ; one of R ^6d or R ^7d is hydrogen and the other is hydroxy protected by hydroxy or C _1-6 alkanoyloxy; or R ^6d and R ^7d together form an oxo group; P is a hydroxy-protected group; M is hydrogen or a metal of group 1A, such as lithium, sodium or potassium; and p, R ⁴ , R ⁵ and R ^x are such as those defined above. The reaction can be carried out in accordance with the procedure disclosed in EP 400971, in which a baccatin derivative III of formula (II), where M is hydrogen, is reacted with azetidinone of formula (III) in the presence of an organic base such as N, N-dimethylaminopyridine. However, it is preferable to first convert the baccatin III derivative to 13-alkoxide by treating it with a strong base such as hydrides, alkylamides and bis (trialkylsilyl) amides of Group 1A metals, as disclosed above in U.S. Patent No. 5,229,526 and Ojima. It is more preferred when the 13-alkoxide is lithium alkoxide. The formation of a lithium salt can be achieved by reacting a compound of formula (II), where M is hydrogen, with a strong metal base such as lithium diisopropylamide, C _1-6 alkyl lithium, lithium bis (trimethylsilyl) amide, phenyl lithium, lithium hydride or the like .

The coupling reaction between a taxane of formula (II) and azetidinone of formula (III) is carried out in an inert organic solvent, such as tetrahydrofuran, at a reduced temperature in the range of about 0 ^° C. to -78 ^° C. Azetidinones of formula (III) can be used as a racemic mixture compatible with the taxane metal oxides of formula (II) in which M is a metal of group IA, in which case the azetidinone reagent is preferably used in an amount of at least 2 equivalents with respect to the taxane reagent, and more preferably about t is about 3 to about 6 equivalents. Chiral azetidinones may also be used, in which case one equivalent of azetidinone relative to taxane may be sufficient, but preferably azetidinone is used in a small excess, for example up to 1.5 equivalents.

The hydroprotective groups may be the same or they may be selected so as to enable the selective removal of one or more protective groups without significantly affecting the other groups; for example, in the compound of formula (Id), R ^2d and PO can both be triethylsilyloxy and R ^3d can be benzyloxycarbonyl; catalytic hydrogenolysis in the presence of palladium on carbon provides the removal of the benzyloxycarbonyl protecting group without removing the triethylsilyl group. Thus, the hydroxy protecting groups of a compound of formula (Id) can be selectively removed to give a compound of formula (Ia).

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

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

В этом случае соединение формулы (IIIa) сначала обрабатывают основанием, такимкакн-бутиллитийилитриэтиламин,азатемсоединениемформулыR⁴(O)_pCO-L, где L -уходящая группа, и в результате получают соединение формулы (III).

In this case, the compound of formula (IIIa) is first treated with a base, such as butyl lithium or triethylamine, and then by the addition of the formula R ⁴ (O) _p CO-L, where L is a leaving group, and the result is a compound of formula (III).

Compounds of formula (IIIa) can be prepared according to the general method disclosed in EP 400971, via the -3-acetoxy-4-substituted-2-azetidinone (IIIb) intermediate, or by the method disclosed in US Pat. No. 5,229,526, via by the intermediate compound -3-triethylsilyloxy-4-substituted-2-azetidinone. In an improved method, compound (IIIb) can be prepared by condensing acetoxyacetyl chloride with bis-imine, followed by hydrogenolysis or acid cleavage to remove the N-imine group; this method is shown in the scheme below, in which R ^{5 ′} is an optionally substituted aryl or heteroaryl group, such as furyl or thienyl. This method is disclosed in US Patent Application N 08/052434, filed April 23, 1993, which is incorporated herein by reference.

Продуктом (IIIb), получаемым от этих реакций циклоприсоединения, обычно является рацемическая смесь двух цисазетидинонов. Рацемическая смесь может быть разделена традиционными способами, такими как превращение в диастереомеры, дифференциальная адсорбция на колонке, наполненной хиральными адсорбентами, или ферментативно. Например, рацемическая смесь соединений формулы (IIIb) может быть введена в контакт с ферментом, который катализирует гидролиз сложного эфира, например, эстераза или липаза, обеспечивая селективное расщепление 3-ацильной группы одного энантиомера без влияния на другой. (См. Brieva et al., J. Org. Chem., 1993, 58:1068-1075, а также одновременно рассматриваемую заявку N 092170 на патент США, поданную 14 июля 1993 г., заявку N 552041 на европейский патент, опубликованную 29 июля 1993 г.). В соответствии с другим вариантом рацемическая смесь может быть сначала подвергнута катализируемому основанием гидролизу для удаления 3-ацильной группы и образования рацемической смеси соответствующего 3-гидрокси- β- лактама, после чего рацемическую смесь 3- гидрокси β- лактата вводят в контакт с ферментом, способным к катализирующему ацилированию гидроксигруппы, чтобы избирательно ацилировать гидроксигруппу одного энантиомера без влияния на другой. Или же рацемическая смесь 3-гидрокси- β- лактама может быть ацилирована хиральной карбоновой кислотой, и полученная диастереомерная смесь может быть затем разделена известными в данной области техники методами с последующим удалением хирального вспомогательного реагента и получением в результате требуемого энантиомера.

The product (IIIb) obtained from these cycloaddition reactions is usually a racemic mixture of two cisazetidinones. The racemic mixture can be separated by conventional methods, such as conversion into diastereomers, differential adsorption on a column filled with chiral adsorbents, or enzymatically. For example, a racemic mixture of compounds of formula (IIIb) can be contacted with an enzyme that catalyzes the hydrolysis of an ester, such as esterase or lipase, to selectively cleave the 3-acyl group of one enantiomer without affecting the other. (See Brieva et al., J. Org. Chem., 1993, 58: 1068-1075, as well as U.S. Patent Application No. 092170, filed July 14, 1993; European Patent Application No. 552041, published 29 July 1993). In another embodiment, the racemic mixture may be first subjected to base-catalyzed hydrolysis to remove the 3-acyl group and form the racemic mixture of the corresponding 3-hydroxy-β-lactam, after which the racemic mixture of 3-hydroxy β-lactate is contacted with an enzyme capable of catalyzing the acylation of a hydroxy group to selectively acylate the hydroxy group of one enantiomer without affecting the other. Alternatively, the racemic mixture of 3-hydroxy-β-lactam can be acylated with a chiral carboxylic acid, and the resulting diastereomeric mixture can then be separated by methods known in the art, followed by removal of the chiral auxiliary reagent and thereby obtaining the desired enantiomer.

In materials Ojima et al., J. Org. Chem., 56: 1681-1683, 1991; Tet. Lett. , 33: 5737-5740, 1992, and Tetrahedron, 48: 6985-7012, 1992, describe the synthesis of a number of chiral azetidinones of formula (IIIa) and / or the corresponding N- (p-methoxyphenyl) -relative, where P is a hydroxy-protecting group of triisopropylsilyl; and R ⁵ -4-methoxyphenyl, 3,4-dimethyloxyphenyl, phenyl, 4-fluorophenyl, 4-trifluoromethylphenyl, 2-furyl, 2-phenylethenyl, 2- (2-furyl) ethenyl, 2-methylpropyl, cyclohexylmethyl, isopropyl, phenethyl , 2-cyclohexylethyl or n-propyl. Parts of these reference materials relevant to this application are hereby incorporated by reference. Other azetidinones within the definition of formula (III), but not specifically disclosed in the referenced materials, may be obtained by those skilled in the art by other methods well known in the art.

Biological assessment
Compounds in accordance with the present invention are new antitumor substances; the present compounds of formula (A) were evaluated in vitro cytotoxicity assays and in vivo animal tumor models.

In vitro cytotoxicity data
The compounds of the present invention showed cytotoxic activity in vitro against cancer cells in the human colon HCT-116 and HCT-116 / VM46. HCT-116 / VM46 cells are cells that were previously selected for resistance to teniposide and express a phenotype that is resistant to many drugs, including resistance to paclitaxel. Cytotoxicity was evaluated in human colon cancer cells HCT-116 by XTT testing (2,3-bis (2-methoxy-4-nitro-5-sulfenyl) -5 - [(phenylamino) carbonyl] - 2H-tetrazolium hydroxide) as described by DA Scudiero et al., "Evaluation of 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 seeded at 4000 cells / well on 96-well microtiter plates, and after 24 hours, drugs were added and diluted sequentially. Cells were incubated at 37 ^° C. for 72 hours, with tetrazolium dye (XTT) being added during this period. The dehydrogenase enzyme in living cells reduces XTT to a form that absorbs light at 450 nm, which can be quantified spectrophotometrically. The greater the uptake, the greater the number of living cells. The results were expressed in terms of IC ₅₀ / drug concentration necessary to inhibit the rapid growth of cells (i.e., absorption at 450 nm) to 50% of the growth of untreated control cells. The IC ₅₀ values for the compounds to be evaluated in this test are given in table 1 at the end of the description.

The 7-O-methylthiomethylpaclitaxel compound (Example 1 (a)), which showed an IC ₅₀ = 0.003 μM against HCT-116 and 0.025 μM against HCT-116 / VM46, was also tested in a cytotoxicity test.

In vivo antitumor activity
Hybrid Balb / cx DBA ₂ F ₁ (CDF ₁ ) mice were implanted subcutaneously (sc) with 0.1 ml of 2% (weight / volume) M109 lung cancer (as described by W.Rose "Evaluation of Madison 109 Lung Carcinoma as a Model for Screening Antitumor Drugs ", Cancer Treatment Reports, 65, N 3-4 pp. 299-312, 1981). The test compounds and the standard drug (paxitaxel) were administered intravenously to groups of mice, each group receiving a compound with a different dose level, as a result of which three to four different doses per compound were evaluated. To assess the survival of mice, they were monitored daily until their death or approximately 75 days after tumor implantation if they did not die. One group of mice per experiment was left untreated for control. Tumors were measured once or twice a week, and the size in millimeters was used to estimate the mass of the tumor in accordance with the published procedure (ibid.).

 The average survival time of compound-treated (T) mice was compared with the average survival time of parallel, control (C) mice. The ratio of two values for each group of mice treated with the compounds 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 treated groups and the average time for the control group with tumor growth to 1 g (expressed as T-C in days). The larger the T-C value, the greater the growth retardation of the primary tumor. Compounds showing% T / C ≥ 125% and / or T-C ≥ 4.0 days are considered active in the model (sample) M109 SC.

 The compound of Example 3 (triethanolamine salt) was then evaluated in mouse and human xenograft tumor models (M109, A2780 / s DDP — human ovarian cancer resistant to cisplatin, and HCT-116 — human colon cancer) compared to paclitaxel as a positive control experience. Model A2780 / s DDP is described in Rose and Basler, In vivo, 1989, 3: 249-254. M109 was subcultured (passaged) subcutaneously every two weeks in Balb / C mice and implanted subcutaneously in CDF1 mice to evaluate antitumor activity. A2780 / s DDP and HCT-116 were grown in goiter-free mice both for reseeding (every two to three weeks) and for therapeutic (therapeutic) experiments. The compound of Example 3 was administered intravenously in water or orally in water with a few drops of Tween 80, and paclitaxel was either suspended in water with Tween 80, or dissolved in cremophor / ethanol (50% / 50%) and diluted with saline. The test regimen with M109 (implantation subcutaneously) was performed once a day for 5 consecutive days, starting from 4 days after tumor implantation. In tests with xenograft of a human tumor, the compounds were administered 5 times - once a day every other day, starting from the moment when the tumors were at a stage of 50-100 mg by weight.

 In one experiment with M109, the compound of Example 3 administered intravenously provided a max. % T / C = 155 (T-C = 19 days) at 36 mg / kg / injection. (for comparison, paclitaxel provided max.% T / C = 132 (T-C = 13 days) at 36 or 18 mg / kg / injection.). In the same experiment, the compound of Example 3 administered orally provided a max. % T / C = 158 (T-C = 22.2 days) at a dose of 160 mg / kg / inject., While paclitaxel at the same dose (the highest of the tested), suspended in water and tween 80 showed no activity. In another experiment with M109, the intravenous compound of Example 3 gave max. % T / C = 170 (T-C = 17 days) at 48 mg / kg / injection. (for comparison, paclitaxel for max.% T / C = 167 (T-C = 14 days) at 48 or 36 mg / kg / injection.). In the same experiment, orally administered compound 3 gave max. % T / C = 172 (T-C = 17 days) at a dose of 200 mg / kg / inject., And paclitaxel dissolved in cremophor / ethanol / saline did not show activity at 60 mg / kg / inject. In this experiment, paclitaxel, dissolved in cremophor / ethanol / saline, at a dose of more than 60 mg / kg / injection. not administered due to limitations due to solubility and toxicity.

 In an experiment with A2780 / c DDP, the intravenously administered compounds of Example 3 showed a max. T-C value = 29.8 days at 36 mg / kg / injection. (cf. paclitaxel gave max. T-C = 26.3 days at 36 mg / kg / injection). The orally administered compound of Example 3 gave max. T-C = 20 days at a dose of 160 mg / kg / inject. When experimenting with HCT-116, treatment with polytaxel at an intravenous dose of 24 or 36 mg / kg / injection. gave 6 cures of 7 or 6 of 8 (respectively) treated mice, and oral administration of the compound of Example 3 at 160 or 240 mg / kg / injection. cured, respectively, 6 or 7 of the 8 treated mice. A cure means the absence of a tumor on day 80 after implantation.

 Compounds in accordance with the present invention are phosphonooxymethyl esters of taxane derivatives. Formulations of pharmaceutically acceptable salts exhibit improved solubility in water compared to paclitaxel, which enables convenient pharmaceutical preparation of dosage forms. Without being bound by theory, the phosphonooxomethyl esters in accordance with the present invention are thought to be drug precursors (prior drug raw materials) of paclitaxel or a derivative thereof, the phosphonooxymethyl fragment being cleaved upon contact with phosphatase in vivo to form a related compound. As shown above, the compounds of the present invention are effective tumor growth retardants. Therefore, the present invention also relates to a method for suppressing tumors in a mammalian organism, which (method) comprises introducing into the body a tumor-bearing host of an effective anti-tumor amount of a compound of formula (A).

The compounds of formula (A) in accordance with the present invention can be used in the same way as paclitaxel, therefore, an oncologist who is a specialist in the treatment of cancer, can prescribe without appropriate experimentation the appropriate treatment course for the administration of the compounds in accordance with the present invention. The dosage, method and schedule of administration of the compounds in accordance with the present invention are not particularly limited and may vary with each particular compound used. Thus, the compound in accordance with the present invention can be introduced by any suitable method, preferably parenterally; the dose may, for example, be in the range of about 1-100 mg / kg body weight, or about 20 to 500 mg / m ² . The compounds of formula (A) can also be administered orally, with the oral dose being in the range of about 5-500 mg / kg body weight. The specific dose used will vary depending on the particular formulation, the route of administration and the specific location, host and type of tumor to be treated. When determining the dose, many factors affecting the effect of the drug are taken into account, such as age, weight, gender, diet and physical condition of the patient.

 The present invention also provides pharmaceutical compositions comprising an anti-tumor amount of a compound 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 US 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, injection forms, 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 reconstituted with sterilized water, physiological saline or a mixture of water and an organic solvent such as propylene glycol, ethanol and the like, or some other sterile injectable medium immediately before use for parenteral administration.

 Typical pharmaceutically acceptable carriers are, for example, mannitol, urea, dextrans, lactose, potato and maize starches, magnesium stearate, talc, vegetable oils, polyalkylene glycols, ethyl cellulose, poly / vinylpyrrolidone /, calcium carbonate, ethyl oleate, isopropyl myristate, benzyl carbonate, benzyl carbonate, benzyl gelatin, potassium carbonate, silicic acid. The pharmaceutical preparation may also contain non-toxic auxiliary substances, such as emulsifying, preserving, wetting agents, etc., such as, for example, sorbitan monolaurate, triethanolamine oleate, polyoxyethylene monostearate, glyceryl tripalmitate, sodium dioctyl sulfosuccinate and the like.

In the following experimental procedures, all temperatures, when there is no specific indication, are presented in ^o C. The spectral characteristics of nuclear magnetic resonance (NMR) refer to chemical shifts (δ), expressed in parts per million (ppm), in comparison with tetramethylsilane ( TMS) as ethanol for comparison. Mentioned relative area for various shifts in the spectral data of proton NMR corresponds to the number of hydrogen atoms of a particular functional type in the molecule. The character of the multiplicity shifts is indicated as a wide singlet (bs), a wide doublet (bd), a wide triplet (pcs), a wide quartet (bcc), a singlet (s), a multiplet (m), a doublet (d), a quartet (sq), triplet (t), doublet of doublets (dd), doublet of triplets (dt), and doublet of quartets (dkv). The solvents used to obtain the NMR spectrum are acetone-d ₆ (deuterated acetone), DMSO-d ₆ (perdeuterodimethyl sulfoxide), D ₂ O (deuterated water), CDCl ₃ (deuterochloroform) and other conventional deuterated solvents. The description of the infrared spectrum (IR spectrum) includes only wave numbers (cm ^-1 ) of absorption having identification values of functional groups.

 Celite is a registered trademark of f. Jones-Manville Products Corporation for Diatomaceous Earth.

 Abbreviations used throughout this specification are conventional abbreviations widely used in the art. Some of them: MS (mass spectrometry); HRMS (high resolution mass spectrometry); Ac (acetyl); Ph (phenyl); v / v (volume / volume, vol / vol); FAB (fast atom bombardment); NOBA (meta-nitrobenzyl alcohol); min (min / s); h (hour / s); NIS (N-iodosuccinimide); BOC (tert-butoxycarbonyl); CBZ (benzyloxycarbonyl); Bn (benzyl); Bz (benzoyl); TES (triethylsilyl); DMSO (dimethyl sulfoxide); THF (tetrahydrofuran); GMDS (hexamethyldisilazane).

Getting raw (source) materials
The following are examples of the preparation of several specific raw materials suitable for the preparation of compounds of formula (A).

(а) 2'-7-О-бис(2,2,2-трихлорэтоксикарбонил)-10-дезацетилпаклитаксел
10-Дезацетилпаклитаксел (140 мг, 0,173 ммоль) в сухом дихлорметане (3,5 мл) обрабатывали при 0^oC пиридином (0,028 мл, 0,346 ммоль) и трихлорэтилхлорформиатом (0,0724 мл, 0,260 ммоль). Через 1 час при этой температуре убирали холодную ванну, и смесь перемешивали при комнатной температуре всю ночь. Выпаривали растворитель, и остаток хроматографировали на силикагеле (30-50% этилацетата в гексане), получив в результате указанное в заголовке соединение в виде пены (92,3 мг, 46%). Дальнейшее элюирование дало непрореагировавший исходный материал (35 мг, 25%) и 2',10-О-бис/2,2,2-трихлорэтоксикарбонил)-10-дезацетилпаклитаксел с выходом 16%.Getting 1. 10-Desacetoxypaclitaxel

(a) 2'-7-O-bis (2,2,2-trichloroethoxycarbonyl) -10-deacetylpaclitaxel
10-Deacetylpaclitaxel (140 mg, 0.173 mmol) in dry dichloromethane (3.5 ml) was treated at 0 ^° C. with pyridine (0.028 ml, 0.346 mmol) and trichloroethyl chloroformate (0.0724 ml, 0.260 mmol). After 1 hour, a cold bath was removed at this temperature, and the mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was chromatographed on silica gel (30-50% ethyl acetate in hexane) to give the title compound as a foam (92.3 mg, 46%). Further elution yielded unreacted starting material (35 mg, 25%) and 2 ', 10-O-bis / 2,2,2-trichloroethoxycarbonyl) -10-deacetylpaclitaxel in 16% yield.

(b) 2 ', 7-O-bis (2,2,2-trichloroethoxycarbonyl) -10-deacetoxy-11,12-dihydropaclitaxel-10,12 (18) -diene
The product obtained in stage (a) (92.3 mg, 0.079 mmol) in dry dichloromethane (2 ml) was treated at room temperature with 1,1,2-trifluoro-2-chlorotriethylamine (0.0384 ml, 0.238 mmol). The solution was stirred overnight. The solvent was evaporated and the residue was purified by column chromatography (25% ethyl acetate in hexane), whereby the title compound was obtained as a white powder (42.8 mg, 47.3%).

(c) 10-Desacetoxy-11,12-dihydropaclitaxel-10,12 (18) -diene
The product obtained in step (b) (39 mg, 0.034 mmol) was dissolved in methanol (0.5 ml) and acetic acid (0.5 ml) and treated with acid washed with zinc dust (66.4 mg, 1.020 mmol). The suspension was heated at 40 ^° C. for 1 hour, filtered, and the filtrate was evaporated. Chromatography of the residue with 60% ethyl acetate / hexane gave the title compound as a foam (22 mg, 81%).

(d) 10-desacetoxypaclitaxel
The product obtained in step (c) (22 mg, 0.028 mmol) in ethyl acetate (0.7 ml) was hydrogenated 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, filtration (washing with ethyl acetate), evaporation and chromatography (60% ethyl acetate in hexane) gave the title product (15.0 mg, 68%) as a white foam.

(a) 2'-O-Бензилоксикарбонил-7-дезокси-7 α- фторпаклитаксел
Диэтиламиносеры трифторид (ДАСТ, 18,7 мкл, 0,141 ммоль) растворяли в сухом дихлорметане (0.5 мл), и этот раствор охлаждали до 0^oC. Добавляли раствор 2'-O-(бензилоксикарбонил)паклитаксела (71 мг, 0,072 ммоль) в дихлорметане (1 мл), и полученный раствор держали при 0^oC 30 мин и при комнатной температуре 4 ч. Затем к реакционной смеси добавляли воду (0,15 мл), чтобы быстро прекратить реакцию, и полученную смесь концентрировали до получения остатка. Остаток хроматографировали на колонке с силикагелем (с элюированием 40% этилацетата в гексане) и получили 61 мг (выход 85,7%) смеси (1: 1) указанного в заголовке соединения и 2'-O-бензилоксикарбонил-8-дезметил-7,8-циклопропапаклитаксела.Getting 2. 7-Deoxy-7α-fluoropaclitaxel

(a) 2'-O-Benzyloxycarbonyl-7-deoxy-7 α- fluoropaclitaxel
Diethylamino sulfur trifluoride (DAST, 18.7 μl, 0.141 mmol) was dissolved in dry dichloromethane (0.5 ml), and this solution was cooled to 0 ^° C. A solution of 2'-O- (benzyloxycarbonyl) paclitaxel (71 mg, 0.072 mmol) in dichloromethane (1 ml), and the resulting solution was kept at 0 ^° C. for 30 minutes and at room temperature for 4 hours. Then water (0.15 ml) was added to the reaction mixture to quickly terminate the reaction, and the resulting mixture was concentrated to a residue. The residue was chromatographed on a silica gel column (eluting with 40% ethyl acetate in hexane) to give 61 mg (85.7% yield) of a 1: 1 mixture of the title compound and 2'-O-benzyloxycarbonyl-8-desmethyl-7, 8-cyclopropapaclitaxel.

(b) 7-Deoxy-7α- fluoropaclitaxel
The product mixture obtained in stage (a) (89 mg) was dissolved in ethyl acetate (3 ml) and the mixture was stirred in a hydrogen medium under pressure slightly above one atmosphere in the presence of palladium on charcoal (10% Pd, 29 mg, 0.027 mmol). After 12 hours, the solvent was removed and the residue was purified by silica gel chromatography (eluting with 40% ethyl acetate in hexane) to give 67.7 mg of the title compound together with 8-desmethyl-7.8-cyclopropaclitaxel.

 The HPLC (high pressure liquid chromatography) method was then used to separate 7-deoxy-7α-fluoropaclitaxel and 8-desmethyl-7.8-cyclopropapaclitaxel.

В сухую колбу в инертной атмосфере добавляли 2'-O-(бензилоксикарбонил)паклитаксел (4 г, 4 ммоль) и сухой толуол (80 мл). Полученную суспензию перемешивали при температуре окружающей среды с одновременным добавлением по каплям сухого тетрагидрофурана, пока не получали в результате бесцветный раствор. Этот раствор охлаждали до -78^oC в ванне с сухим льдом и ацетоном и затем обрабатывали диэтиламиносеры трифторидом (ДАСТ, 1,2 мл, 2,5 эквив). Реакционной смеси позволяли перемешиваться в течение 16 ч, когда она постепенно нагревалась до температуры окружающей среды. Полученную суспензию фильтровали и фильтрат (разбавленный 30 мл этилацетата) промывали насыщенным водным раствором бикарбоната натрия и затем рассолом. Органическую фракцию высушивали (MgSO₄) и концентрировали, получив в результате неочищенный (сырой) продукт в виде белой пены. Неочищенный материал частичного очищали путем хроматографии на колонке с силикагелем (с элюированием 10% CH₃CN в CH₂Cl₂) и получали 1,45 г смеси 2'-O-(бензилоксикарбонил)-7-дезокси -7α- фторпаклитаксела и 2'-O-(бензилоксикарбонил)-8-дезметил-7,8-циклопропапаклитаксела (смесь 82:18 по ¹H-ЯМР).Equipment
Pump: PE Series 4
Column: Chandon hypercarb (graphitized carbon), 7 microns, 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
Terms
Mobile phase: 85:15 methylene chloride: hexane
The separation did not disappear at 80: 19: 1 methylene chloride: hexane: isopropyl alcohol
Flow rate: 2.5 ml / min
Detector: 254 nm
Thinner: The sample was dissolved in methylene chloride.
Get 3. 7-Deoxy-7α- fluorobaccatin III

In a dry flask in an inert atmosphere, 2'-O- (benzyloxycarbonyl) paclitaxel (4 g, 4 mmol) and dry toluene (80 ml) were added. The resulting suspension was stirred at ambient temperature while dry tetrahydrofuran was added dropwise until a colorless solution was obtained. This solution was cooled to -78 ^° C in a bath of dry ice and acetone, and then diethylamino sulfur was treated with trifluoride (DAST, 1.2 ml, 2.5 equiv). The reaction mixture was allowed to stir for 16 hours, when it was gradually heated to ambient temperature. The resulting suspension was filtered and the filtrate (diluted with 30 ml ethyl acetate) washed with saturated aqueous sodium bicarbonate and then brine. The organic fraction was dried (MgSO ₄ ) and concentrated, resulting in a crude (crude) product as a white foam. Partial crude material was purified by silica gel column chromatography (eluting with 10% CH ₃ CN in CH ₂ Cl ₂ ) to obtain 1.45 g of a mixture of 2'-O- (benzyloxycarbonyl) -7-deoxy-7α-fluoropaclitaxel and 2 ' -O- (benzyloxycarbonyl) -8-desmethyl-7,8-cyclopropapaclitaxel (82:18 mixture of ¹ H-NMR).

The above mixture (1.45 g) was dissolved in ethyl acetate (60 ml) and treated with palladium-carbon (300 mg). After shaking for 4 hours in a hydrogen atmosphere under a pressure of 50 psi (3.5 kg / cm ² ), the reaction mixture was communicated with the atmosphere and filtered through a short column of silica gel and concentrated. This gave a mixture of the desired products — 7-deoxy-7α-fluoropaclitaxel and 8-desmethyl-7.8-cyclopropapaclitaxel — as a white foam (1.24 g, 99% yield, 90:10 mixture by ¹ H-NMR). This mixture was dissolved in dry methylene chloride (30 ml) and treated with tetrabutylammonium borohydride (745 mg, 2.9 mmol, 2 equiv.) And allowed to stir for 6 hours. Then the reaction mixture was quickly cooled with acetic acid (1 ml), diluted with additional methylene chloride ( 30 ml) and washed with saturated aqueous sodium bicarbonate. The organic fraction was dried (MgSO ₄ ) and concentrated. The crude mixture (substituted taxon nuclei) was partially purified by chromatography on a column of silica gel) eluting with 10% CH ₃ CN in CH ₂ Cl ₂ ) and the result was a mixture with a ratio of 90:10 (as determined by ¹ H-NMR) 7- deoxy-7 α-β-fluorobaccatin III and 8-desmethyl-7,8-cyclopropabaccatin III (510 mg, 60%) as a white foam. The resulting foam was crystallized from hot isopropanol, resulting in 7-deoxy-7α-fluorobaccatin III (in the form of small white needles with a yield of 410 mg); so pl. 234 - 236 ^o C (decomposition).

(a) 2'-O-Бензилоксикарбонил-10-дезацетоксипаклитаксел
10-Безацетоксипаклитаксел (27 мг, 0,034 ммоль) в дихлорметане (1 мл) обрабатывали бензилхлорформиатом (0,0146 мл, 0,102 ммоль) и затем диизопропилэтиламином (0,0177 мл, 0,102 ммоль). Реакционную смесь перемешивали при 0^oC 45 мин и при комнатной температуре в течение 12 ч. Выпаривание растворителя и хроматографии на колонке с силикагелем (с элюированием 40% этилацетата в гексане) дали 25,5 мг (выход 81%) указанного в заголовке соединения в виде пены.Preparation 4. 10-Deacetoxy-7-deoxy-7α-fluoropaclitaxel

(a) 2'-O-benzyloxycarbonyl-10-deacetoxypaclitaxel
10-Bezacetoxypaclitaxel (27 mg, 0.034 mmol) in dichloromethane (1 ml) was treated with benzyl chloroformate (0.0146 ml, 0.102 mmol) and then with diisopropylethylamine (0.0177 ml, 0.102 mmol). The reaction mixture was stirred at 0 ^° C. for 45 minutes and at room temperature for 12 hours. Evaporation of the solvent and silica gel column chromatography (eluting with 40% ethyl acetate in hexane) gave 25.5 mg (81% yield) of the title compound in in the form of foam.

(b) 10-Deacetoxy-7-deoxy -7α- fluoropaclitaxel
The product obtained in stage (a) (25.5 mg, 0.028 mmol) in dichloromethane (0.8 ml) was treated with DAST (0.0071 ml, 0.055 mmol). After 45 minutes at 0 ^° C., the reaction was allowed to continue for 5 hours at room temperature. Evaporation of the solvent and chromatography gave 2'-O-benzyloxycarbonyl-7-deoxy-7α-fluoropaclitaxel as a crude foam. This compound was dissolved in ethyl acetate (1 ml) and stirred in a hydrogen medium at a pressure slightly above one atmosphere in the presence of palladium on charcoal (10%, 8.9 mg) for 12 hours at room temperature. The catalyst was removed by filtration, and the product was chromatographed on a silica gel column, then 10 mg (yield 40% in two steps) of the title product as a foam was obtained.

Раствор 2', 10-O-бис(2,2,2-трихлорэтоксикарбонил)-10-дезацетилпаклитаксела (120 мг, 0,103 ммоль) в дихлорметане (2 мл) охлаждали при 0^oC и обрабатывали диэтиламиносеры трифторидом (0,0266 мл, 0,207 ммоль). Раствор перемешивали при 0^oC 30 мин и при комнатной температуре 4 ч. Реакцию резко прекращали путем добавления воды (0,05 мл). Реакционную смесь концентрировали, и остаток очищали путем хроматографии на колонке с силикагелем (с элюированием 30% этилацетата в гексане), получив в результате 81 мг (выход 68%) 2',10-O-бис(2,2,2-трихлорэтоксикарбонил)-7-дезокси 7α- фторпаклитаксела в виде пены. Это соединение (63 мг, 0,054 ммоль) растворяли в метаноле (0,5 мл) и уксусной кислоте (0,5 мл) и обрабатывали цинковой пылью (104 мг, 1,62 ммоль) в течение 90 мин при 45^oC. Реакционную смесь фильтровали, и фильтрат концентрировали. Хроматография на силикагеле (с элюированием 40% гексана в 60% этилацетата) упомянутого остатка дала 38 мг (выход 86%) указанного в заголовке соединения в виде белого твердого вещества.Preparation 5. 10-Deacetyl-7-deoxy-7α-fluoropaclitaxel

A solution of 2 ', 10-O-bis (2,2,2-trichloroethoxycarbonyl) -10-deacetylpaclitaxel (120 mg, 0.103 mmol) in dichloromethane (2 ml) was cooled at 0 ^° C and treated with diethylaminosulfur trifluoride (0.0266 ml, 0.207 mmol). The solution was stirred at 0 ^° C. for 30 minutes and at room temperature for 4 hours. The reaction was abruptly stopped by adding water (0.05 ml). The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (eluting with 30% ethyl acetate in hexane) to obtain 81 mg (68% yield) 2 ', 10-O-bis (2,2,2-trichloroethoxycarbonyl) -7-deoxy 7α-fluoropaclitaxel as a 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, 1.62 mmol) for 90 minutes at 45 ^° C. the mixture was filtered and the filtrate was concentrated. Silica gel chromatography (eluting with 40% hexane in 60% ethyl acetate) of the above residue gave 38 mg (86% yield) of the title compound as a white solid.

(a) 7-O-[Метилтио)тиокарбонил]баккатин III
Баккатин III (750 мг, 1,278 ммоль) растворяли в сухом тетрагидрофуране (20 мл) и в раствор добавляли одной порцией имидазол (8,7 мг, 0,128 ммоль). Добавляли при комнатной температуре натрия гидрид (50% в минеральном масле, 77 мг, 1,597 ммоль). После прекращения выделения газов (10 мин) добавляли за один раз дисульфид углерода (4,6 мл). Через 3 ч при комнатной температуре желтый раствор обрабатывали метилиодидом (0,238 мл, 3,835 ммоль) и перемешивали всю ночь. Обработка этилацетатом и водой дала указанное в заголовке соединение в виде неочищенного (сырого) масла.Getting 6. 7-Deoxybaccatin III

(a) 7-O- [Methylthio) thiocarbonyl] baccatin III
Baccatin III (750 mg, 1.278 mmol) was dissolved in dry tetrahydrofuran (20 ml) and imidazole (8.7 mg, 0.128 mmol) was added in one portion. Sodium hydride was added at room temperature (50% in mineral oil, 77 mg, 1.597 mmol). After gas evolution ceased (10 min), carbon disulfide (4.6 ml) was added at a time. After 3 hours at room temperature, the yellow solution was treated with methyl iodide (0.238 ml, 3.835 mmol) and stirred overnight. Treatment with ethyl acetate and water gave the title compound as a crude (crude) oil.

 Alternative procedure.

Baccatin III (394 mg, 0.672 mmol) was dissolved in tetrahydrofuran (5 ml) and carbon disulfide (1 ml). Sodium hydride (40.3 mg, 60%, 1.009 mmol) was added to this solution. A catalytic amount of imidazole was also added. The reaction mixture was stirred at room temperature for 1.5 hours, after which methyl iodide (122.8 μl, 2.016 mmol) was added. After 40 minutes, the solvent was removed in vacuo and the residue was chromatographed on silica gel (eluting with 20% -50% -60% ethyl acetate in hexanes) to give the title compound (260 mg, 57.2% yield) together with 7- epibaccatin (98.5 mg, 25%),
(b) 7-O - [(Methylthio) thiocarbonyl] -13-O-triethylsilyl baccatin III
The product of step (a) as a crude oil was dissolved in dry dimethylformamide (5 ml) and treated with imidazole (870 mg, 12.78 mmol) and triethylsilyl chloride (2.10 ml, 12.78 mmol) at room temperature for 15 hours. Water was added, followed by extraction into ethyl acetate. The organic layer was washed with water for a long time and then dried. Silica gel flash chromatography (eluting with 20% ethyl acetate in hexane) afforded the title compound as a glassy solid (yield 209 mg, 209th step in two steps).

Alternative procedure
The product obtained in stage (a) (193.4 mg, 0.286 mmol) was dissolved in dry dimethylformamide (2.86 ml). Imidazole (77.9 mg, 1.14 mmol) was added to this solution, followed by triethylsilyl chloride (192 μl, 1.14 mmol). The reaction mixture was stirred overnight at room temperature. After 12 hours, 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 vacuo. The residue was chromatographed on silica gel (eluting with 20% ethyl acetate in hexanes) to give the title product (163 mg, 72.0% yield).

(c) 7-Deoxy-13-O-triethylsilylbaccatin III
The product obtained in stage (b) (182 mg, 0.230 mmol) in dry benzene (5 ml) was heated to 80 ^° C. in the presence of tributyltin hydride (0.310 ml, 1.150 mmol) and 2,2'-azobisisobutyronitrile (AIBN, 10 mg) . After 3 hours, the solution was allowed to cool, and the solvent was evaporated in vacuo. Silica gel chromatography (eluting with 20% ethyl acetate in hexane) of the obtained residue gave the title compound as an oil.

(d) 7-Deoxybaccatin III
The product obtained in step (c) was dissolved in tetrahydrofuran (5 ml) and treated with tetrabutylammonium fluoride (1 M in tetrahydrofuran, 0.50 ml, 0.50 mmol) for 2 hours at room temperature. After dilution with ethyl acetate and washing with water and brine, followed by chromatography on silica gel (eluting with a 1: 1 mixture of ethyl acetate and hexane), the title compound was obtained as a white glassy solid (63 mg, 58% yield in two steps).

(a) 10-Дезацетил-10-O-(пентафторфенокси)тиокарбонил-7-O- триэтилсилилбаккатин III
7-O-Триэтилсилил-10-дезацетилбаккатин III (см. Green et. al., J. Am. Chem. Soc 110, стр. 5917, 1988) (319 мг, 0,485 ммоль) растворяли в сухом тетрагидрофуране (5 мл), охлаждали до -40^oC и обрабатывали н-бутиллитием (1,58 М в гексанах, 0,384 мл, 0,606 ммоль). Через 40 минут при этой температуре добавляли с помощью шприца неразбавленный пентафторфенилхлортиоформиат (0,086 мл, 0,536 ммоль). Реакционную смесь перемешивали при -20^oC в течение 90 мин, быстро охлаждали насыщенным раствором аммонийхлорида и экстрагировали этилацетатом. Слой этилацетата высушивали и концентрировали. Остаток очищали путем хроматографии на силикагеле (с элюированием 40% этилацетата в гексане) и получили в результате указанное в заголовке соединение в виде пены (320 мг, выход 74%).Getting 7. 10-Desacetoxybaccatin III

(a) 10-Desacetyl-10-O- (pentafluorophenoxy) thiocarbonyl-7-O- triethylsilyl baccatin III
7-O-Triethylsilyl-10-deacetylbaccatin III (see Green et. Al., J. Am. Chem. Soc 110, p. 5917, 1988) (319 mg, 0.485 mmol) was dissolved in dry tetrahydrofuran (5 ml), cooled to -40 ^° C and treated with n-butyl lithium (1.58 M in hexanes, 0.384 ml, 0.606 mmol). After 40 minutes at this temperature, undiluted pentafluorophenylchlorothioformate (0.086 ml, 0.536 mmol) was added via syringe. The reaction mixture was stirred at -20 ^° C. for 90 minutes, quickly cooled with saturated ammonium chloride and extracted with ethyl acetate. The ethyl acetate layer was dried and concentrated. The residue was purified by silica gel chromatography (eluting with 40% ethyl acetate in hexane) to give the title compound as a foam (320 mg, 74% yield).

(b) 10-Desacetoxy-7-O-triethylsilylbaccatin III
The product obtained in stage (a) (119 mg, 0.135 mmol) was dissolved in dry toluene (3 ml) and treated with AIBN-ohm (2 mg). The solution was degassed with dry nitrogen and then tributyltin hydride (0.055 ml, 0.202 mmol) was added. After that, the solution was heated at 90 ^° C. for 1 hour. The solvent was then evaporated and the residue was chromatographed on silica gel (eluting with 40% ethyl acetate in hexane) to give the title compound (87 mg, 99% yield) as a colorless foam.

(c) 10-Desacetoxybaccatin III
The product obtained in step (b) (120 mg, 0.186 mmol) was dissolved in acetonitrile (3.5 ml), and the solution was cooled to -10 ^° C. Concentrated HCl (36%, 0.060 ml) was added and the solution was stirred for 30 minutes. 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 (eluting with 70% ethyl acetate in hexane) to give 10-deacetyloxybaccatin III as a foam (75 mg, 76% yield).

(a) 7-O[(Метилтио)тиокарбонил]-10-дезацетоксибаккатин III
10-дезацетоксибаккатин III (75 мг, 0,142 ммоль) растворили в сухом тетрагидрофуране (2 мл) и дисульфиде углерода (0,5 мл). Затем добавляли гидрид натрия (60% в минеральном масле, 8,5 мг, 0,213 моль), и смесь перемешивали при комнатной температуре в течение 2 ч. Добавляли иодметан (0,026 мл, 0,426 ммоль), и позволяли реакции продолжаться всю ночь. Затем удаляли растворитель, и остаток очищали путем хроматографии на силикагеле (с элюированием 50 - 70% этилацетата), получив в результате указанное в заголовке соединение в виде пены (46,6 мг, выход 53%).Preparation 8. 10-Desacetoxy-7-deoxybaccatin III

(a) 7-O [(Methylthio) thiocarbonyl] -10-deacetoxybaccatin III
10-desacetoxybaccatin III (75 mg, 0.142 mmol) was dissolved in dry tetrahydrofuran (2 ml) and carbon disulfide (0.5 ml). Sodium hydride (60% in mineral oil, 8.5 mg, 0.213 mol) was then added, and the mixture was stirred at room temperature for 2 hours. Iodomethane (0.026 ml, 0.426 mmol) was added and the reaction allowed to continue overnight. Then the solvent was removed, and the residue was purified by silica gel chromatography (eluting with 50-70% ethyl acetate), whereby the title compound was obtained as a foam (46.6 mg, 53% yield).

(b) 10-Desacetoxy-7-deoxybaccatin III
The product obtained in stage (a) (36 mg, 0.058 mmol) in benzene (1 ml) was heated under reflux in the presence of AIBN (2 mg) and tributyltin hydride (0.079 ml, 0.290 mmol) in argon for 3 hours. Concentration the reaction mixture and silica gel flash chromatography of the residue (eluting with 40% ethyl acetate in hexanes) followed by separation by HPLC (high pressure liquid chromatography) from the other components gave the title compound as a foam (16.8 mg, 56% yield) .

Alternative procedure
To a solution of 7-O - [(methylthio) thiocarbonyl] -13-O-triethylsilylbaccatin III (product of Preparation 6, step (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 degassed with dry N ₂ for 5 minutes Tributyltin hydride (708.7 μl, 2.63 mmol) was added, and the reaction mixture was heated at 100 ^° C for 2 hours, after which another portion of tributyltin hydride (425.3 μl, 1.581 mmol) was added. The reaction mixture was heated for 5.5 hours at 100 ^° C., after which it was allowed to cool to room temperature. Silica gel chromatography (eluting with 20% ethyl acetate in hexanes) gave 7-deoxy-10-deacetoxy-13-O- (triethylsilyl) baccatin III (320 mg, 97% yield).

 To a solution of the product obtained in the above step (160 mg, 0.255 mmol) in dry tetrahydrofuran (2 ml) was added tetrabutylammonium fluoride (766 μl, 1 M, 0.766 mmol) at room temperature. The reaction mixture was stirred for 1 h at room temperature. The solvent was removed and the residue was chromatographed on silica gel (eluting with 50-70% ethyl acetate in hexanes) to give the title target product (115 mg, 87.9% yield).

К перемешанному раствору (3R,4S)-4-фенил-3-триэтилсилилокси-2-азетидинона (2,200 г, 7,92 ммоль) в сухом тетрагидрофуране (25 мл) N,N-диизопропилэтиламин (1,65 мл, 9,510 ммоль, 1,2 эквив.) при 0^oC в среде аргона. Раствор перемешивали 5 мин, после чего добавляли ди-трет-бутилдикарбонат (2,080 г, 9,510 ммоль, 1,2 эквив.) и 4-диметиламинопиридин (193,6 мг, 1,581 ммоль, 0,20 эквив. ). Реакционную смесь перемешивали при 0^oC 60 мин и затем разбавляли этилацетатом (25 мл). Полученный раствор промывали рассолом, 10%-ным NaHCO₃, 10%-ным раствором HCl, высушивали (MgSO₄) и концентрировали, получив в результате неочищенное соединение (масло). Это соединение затем очищали путем флеш-хроматографии на силикагеле (с элюированием 15% этилацетата в гексанах) и получили указанное в заголовке соединение в виде белого твердого вещества (2,4 г, выход 83%).Preparation 9. (3R, 4S) -1-tert-Butoxycarbonyl-4-phenyl-3-triethylsilyloxy-2-azetidinone

To a mixed solution of (3R, 4S) -4-phenyl-3-triethylsilyloxy-2-azetidinone (2.200 g, 7.92 mmol) in dry tetrahydrofuran (25 ml) N, N-diisopropylethylamine (1.65 ml, 9.510 mmol, 1.2 equiv.) At 0 ^o C in argon. The solution was stirred for 5 minutes, after which di-tert-butyl dicarbonate (2.080 g, 9.510 mmol, 1.2 equiv.) And 4-dimethylaminopyridine (193.6 mg, 1.581 mmol, 0.20 equiv.) Were added. The reaction mixture was stirred at 0 ^° C. for 60 minutes and then diluted with ethyl acetate (25 ml). The resulting solution was washed with brine, 10% NaHCO ₃ , 10% HCl, dried (MgSO ₄ ) and concentrated, resulting in a crude compound (oil). This compound was then purified by flash chromatography on silica gel (eluting with 15% ethyl acetate in hexanes) to give the title compound as a white solid (2.4 g, 83% yield).

(a) В трехгорлую круглодонную колбу емкостью 1 л, снабженную термометром, магнитной мешалкой и капельной воронкой, добавляли гидробензамид (30,00 г, 100,5 ммоль) и этилацетат (150 мл). Перемешивая под защитным слоем аргона, реакционную смесь охлаждали до 5^oC и добавляли к ней триэтиламин (16,8 мл, 121 ммоль). Затем добавляли по каплям в течение 90 мин раствор ацетоксиацетилхлорида (12,4 мл, 116 ммоль) в этилацетате (300 мл). Через 16 часов при этой температуре реакционной смеси давали возможность нагреваться до 20^oC (1,5 ч) и переносили ее в делительную воронку. Органический слой промывали последовательно насыщенным водным раствором NH₄Cl /150 мл, 100 мл/, насыщенным водным раствором NaHCO₃ (120 мл) и рассолом (120 мл). Для целей идентификации указанное в заголовке соединение может быть выделено на этой стадии путем высушивания органической фазы над MgSO₄, фильтрования и удаления растворителя в вакууме. Это дало (±)-цис-3-ацетилокси-1-[(фенил) (бензилиденимино)метил]-4-фенилазетидин-2-он с количественным выходом сырого продукта в виде красного стекла.Getting 10. (±) -cis-3-Acetoxy-4-phenylazetidin-2-one

(a) Hydrobenzamide (30.00 g, 100.5 mmol) and ethyl acetate (150 ml) were added to a 1 L three-necked round bottom flask equipped with a thermometer, a magnetic stirrer and a dropping funnel. Stirring under a protective layer of argon, the reaction mixture was cooled to 5 ^° C. and triethylamine (16.8 ml, 121 mmol) was added thereto. Then, a solution of acetoxyacetyl chloride (12.4 ml, 116 mmol) in ethyl acetate (300 ml) was added dropwise over 90 minutes. After 16 hours at this temperature, the reaction mixture was allowed to warm to 20 ^° C. (1.5 hours) and transferred to a separatory funnel. The organic layer was washed successively with saturated aqueous NH ₄ Cl / 150 ml, 100 ml /, saturated aqueous NaHCO ₃ (120 ml) and brine (120 ml). For identification purposes, the title compound can be isolated at this stage by drying the organic phase over MgSO ₄ , filtering and removing the solvent in vacuo. This gave (±) -cis-3-acetyloxy-1 - [(phenyl) (benzylidenimino) methyl] -4-phenylazetidin-2-one with a quantitative yield of the crude product as red glass.

(b) A solution of the compound obtained in part (a) in ethyl acetate (500 ml) was carefully transferred under a stream of argon into a 2 liter Parr flask containing 10% palladium on activated carbon (6.00 g). This mixture was treated with hydrogen (4 atm.) For 20 hours, after which the catalyst was removed by filtration through a celite pad. The filtration residue was suspended in ethyl acetate (200 ml), stirred (10 min) and filtered. The filtration residue was washed with ethyl acetate (100 ml), and the filtrates were combined. The organic layer was washed with 10% HCl (300 ml), and both layers were filtered through a sintered glass funnel to remove a white precipitate (dibenzylamine • HCl), which was washed with ethyl acetate (100 ml). The phases were separated, and the organic layer was washed with another portion of 10% HCl (200 ml). The combined washings with 10% HCl were again extracted with ethyl acetate (200 ml), and the combined organic layers were washed with saturated aqueous NaHCO ₃ (300 ml) and brine (250 ml). The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo to a final volume of 75 ml. This mixture was cooled to 4 ^° C. and the precipitated product was separated by filtration. The filtration residue was washed with hexane (200 ml), resulting in 16.12 g (78.1% of the total yield from hydrobenzamide) of the title compound as white needles; so pl. = 150 - 151 ^o C.

(a) Следовали процедуре, описанной в Получении 10, часть (a), за исключением того, что вместо гидробензамида, использовали гидрофурамид [т.е. 2-фурил-CH-(N= CH-2-фурил)₂] , и реакцию проводили в расчете на 18,6 ммоль (вместо 100 ммоль). В результате гидрофурамид (5,00 г, 18,6 ммоль), триэтиламин (3,11 мл, 22,3 ммоль) и ацетоксиацетилхлорид (2,30 мл, 21,4 ммоль) дали 6,192 г (выход 90,4%) соединения (±)-цис-3-ацетилокси-1-[(2-фурил)(2-фурилметиленимино)-метил] -4- (2-фурил)азетидин-2-он в виде бледно-красного сиропа.Preparation 11. (±) -cis-3-Triethylsilyloxy-4- (2-furyl) -N-tert-butoxycarbonylazetidin-2-one

(a) The procedure described in Preparation 10, part (a) was followed, except that instead of hydrobenzamide, hydrofuramide was used [i.e. 2-furyl-CH- (N = CH-2-furyl) ₂ ], and the reaction was carried out calculated on 18.6 mmol (instead of 100 mmol). As a result, hydrofuramide (5.00 g, 18.6 mmol), triethylamine (3.11 ml, 22.3 mmol) and acetoxyacetyl chloride (2.30 ml, 21.4 mmol) gave 6.192 g (90.4% yield) compounds of (±) -cis-3-acetyloxy-1 - [(2-furyl) (2-furylmethyleneimino) methyl] -4- (2-furyl) azetidin-2-one in the form of a pale red syrup.

(b) The procedure described in Preparation 10, part (b) was followed, except that the product was isolated by preparative thin layer chromatography, and the reaction was carried out based on 2.7 mmol, instead of the initial amount of hydrofuramide. Thus, the crude product obtained in part (a) described above was redissolved in ethyl acetate (50 ml) and added to 10% palladium on activated carbon (150 mg). Purification of the crude solid by preparative thin-layer chromatography (2 mm silica gel, eluted with 1: 1 ethyl acetate-hexane) gave 386 mg (adjusted overall yield of hydrofuramide 65.8%) (±) -cis-3- (acetyloxy) - 4- (2-furyl) azetidin-2-one as a yellow solid. This compound was recrystallized from ethyl acetate-hexane; so pl. = 118 - 119 ^o C.

(c) The compound (3.78 g, 19.4 mmol) obtained in the above-described part (b) in 60 ml of methanol was stirred with K ₂ CO ₃ (20 mg, 0.14 mmol) for 90 minutes, and the solution was neutralized by Dowex 50w-X8 and filtered. The filtrate was concentrated, and the residue was dissolved in 80 ml of anhydrous THF (tetrahydrofuran) and stirred at 0 ^° C. with imidazole (1.44 g, 21.2 mmol) and TESCl (3.4 ml, 20.2 mmol) for 30 minutes . The solution was diluted with ethyl acetate and washed with brine, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (hexane-ethyl acetate 3: 1 eluted) to give 4.47 g (86% yield) of (±) -cis-3-triethylsilyloxy-4- (2-furyl) azetidine-2- it is in the form of a colorless oil.

(d) The product obtained in part (c) (2.05 g, 7.7 mmol) in 30 ml of dichloromethane was stirred at 0 ^° C. with diisopropylethylamine (1.5 ml, 8.6 mmol) and di-tert-butyl dicarbonate ( 2.0 g, 9.2 mmol), as well as a catalytic amount of dimethylaminopyridine (DMAP). The solution was diluted with dichloromethane and washed with brine, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (eluting with 8: 1 hexane-ethyl acetate) to give 2.0 g (70% yield) of the title compound as a waxy solid.

 The racemic mixture obtained in part (b) can be used as a substrate for enzymatic hydrolysis using lipase, such as PS-30 from Pseudomonas sp. (Amano International Co.) to obtain (3R, 4R) -3-hydroxy-4- (2-furyl) -azetidin-2-one. An enzymatic separation method using PD-30 lipase and other enzymes is disclosed in U.S. Patent Application No. 092170, filed by these applicants on July 14, 1993, which is hereby incorporated by reference in its entirety.

 Following the procedures in parts (c) and (d) using (3R, 4R) -3-hydroxy-4- (2-furyl) -azetidin-2-one, (3R, 4R) -N- (tert-butoxycarbonyl) is obtained ) -3-triethylsilyloxy-4- (2-furyl) azetidin-2-one.

(a) Следовали процедуре, описанной в Получении 10, стадия (a), за исключением того, что вместо гидробензамида, использовали гидротиенамид [т.е. 2-тиенил-CH-(N = CH-2-тиенил)₂]. При этом гидротиенамид (30 г, 94,7 ммоль), триэтиламин (15,84 мл, 114 ммоль) и ацетоксиацетилхлорид (11,6 мл, 108 ммоль) дали (±)-цис-3-ацетилокси-1-[(2-тиенил) (2-тиенилметиленимино)метил]-4-(2-тиенил)азетидин-2-он в виде вязкого масла.Preparation 12. (±) -cis-3-Triethylsilyloxy-4- (2-thienyl) -N-tert-butoxycarbonylazetidin-2-one

(a) The procedure described in Preparation 10 was followed, step (a), except that instead of hydrobenzamide, hydrothienamide was used [i.e. 2-thienyl-CH- (N = CH-2-thienyl) ₂ ]. Hydrothienamide (30 g, 94.7 mmol), triethylamine (15.84 ml, 114 mmol) and acetoxyacetyl chloride (11.6 ml, 108 mmol) gave (±) -cis-3-acetyloxy-1 - [(2 thienyl) (2-thienylmethyleneimino) methyl] -4- (2-thienyl) azetidin-2-one as a viscous oil.

(b) To a stirred solution of the product obtained in part (a) (0.431 g, 1.03 mmol) in dichloromethane (2.93 ml) at 25 ^° C was added a 70% aqueous solution of acetic acid (0.35 ml of glacial acetic acid) acid and 0.15 ml of water). The reaction mixture was refluxed and stirred for 2.5 hours. The reaction mixture was diluted with 50 ml of dichloromethane and then washed with two 75 ml portions of a saturated aqueous bicarbonate solution, and then with one 50 ml portion of a saturated brine. The organic extract was concentrated in vacuo to a brown oil, dissolved in a minimum amount of dichloromethane and then placed on a 4 '' x 0.5 '' silica gel column (10.16 cm x 1.27 cm). Elution using a gradient of 10-60% ethyl acetate in hexane gave less polar byproducts and then (±) -cis-3-acetyloxy-4- (2-thienyl) -azetidin-2-one (0.154 g, 75% yield) in as a white solid.

(c) a solution of the product obtained in part (b) (2.5 g, 11.8 mmol) was dissolved in methanol (10 ml) and treated with saturated aqueous sodium bicarbonate (10 ml), and the resulting suspension was allowed to mix at ambient temperature for 3 hours. Then the reaction mixture was diluted with ethyl acetate (20 ml) and washed with water (15 ml). The aqueous fraction was back extracted several times with ethyl acetate, and the combined organic fractions were dried (MgSO ₄ ) and concentrated to give a yellow solid (yield 1.7 g). The crude material was dissolved in dry tetrahydrofuran (20 ml), and the solution was cooled to 5 ^° C. in an ice water bath. After stirring for 5 minutes, triethylchlorosilane (1.85 ml, 1.1 equiv.) Was added dropwise. The resulting suspension was allowed to mix for 3 hours at the indicated temperature, after which the solids were separated by filtration. The organic layer was washed with water (2 x 20 ml), and then dried (MgSO ₄ ) and concentrated. The crude product was purified by silica gel column chromatography (eluted with hexanes-ethyl acetate 7: 3) to give (±) -cis-3-triethylsilyloxy-4- (2-thienyl) azetidin-2-one as a colorless solid substances (1.5 g, yield 45%); etc. 70 - 71 ^o C.

Alternative procedure
The product obtained in part (b) (2.0 g, 9.37 mmol) in 40 ml of methanol was stirred with K ₂ CO ₃ (60 mg, 0.43 mmol) for 30 minutes, and the solution was neutralized with Dowex 50W-X8 and filtered . The filtrate was concentrated, and the residue was dissolved in 50 ml of anhydrous THF and stirred at 0 ^° C 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 MgSO ₄ and concentrated. The residue was chromatographed on silica gel (eluting with 3: 1 hexane-ethyl acetate) to give 2.13 g (86% yield) of the title compound as a colorless oil.

(d) A solution of the product obtained in part (c) (425.7 mg, 1.48 mmol) was dissolved in dichloromethane (10 ml) and cooled to 5 ^° C. in an ice water bath. The reaction mixture was treated with a catalytic amount of DMAP, followed by diisopropylethylamino (TESCl, 0.25 ml, 1.0 equiv.) And then di-tert-butyl dicarbonate (388.4 mg, 1.2 equiv.). After stirring for 2 hours at this temperature, the reaction was quickly terminated with saturated aqueous sodium bicarbonate (45 ml), and the organic fraction was washed with water (5 ml), then dried (MgSO ₄ ), passed through a short plug of silica gel and concentrated to obtain as a result, the target product in the form of a colorless oil (525.3 mg, yield 93%).

 The procedure described above in Preparations 9, 11 (d) and 12 (d) can be adapted to produce other N-substituted azetidinones suitable for preparing the compounds of the present invention. Examples of such azetidines are listed in table III; P is a hydroxy protecting group such as triethylsilyl, triisopropylsilyl and ethoxyethyl.

10-Дезацетокси-7-O-триэтилсилилбаккатин III (100 мг, 0,156 ммоль) помещали в колбу под аргоном и растворяли в сухом тетрагидрофуране (1,5 мл). После охлаждения до -40^oC добавляли по каплям н-бутиллитий (1,45 М в гексанах, 0,119 мл, 0,170 ммоль), а затем (3R, 4S)-1-трет-бутоксикарбонил-4-фенил-3-триэтилсилилокси-2-азетидинон (94,2 мг, 0,25 ммоль) в тетрагидрофуране (0,5 мл) в течение 2 мин. Смесь сразу же нагревали до 0^oC и перемешивали 45 мин, перед тем как резко охладить насыщенным аммонийхлоридом (3 мл). Смесь экстрагировали этилацетатом, высушивали и концентрировали. Хроматография на силикагеле (с элюированием 30% этилацетата в гексане) дала 10-дезокси-2', 7-бис-O-(триэтилсилил)таксотер в виде пены (125 мг, выход 76%). Это соединение (100 мг, 0,098 ммоль) немедленно растворяли в ацетонитриле (2 мл) при -5^oC и обрабатывали хлористоводородной кислотой (0,037 мл, 36%, 12 М). Смесь перемешивали 2 ч при -5^oC, после чего резко охлаждали водным бикарбонатом, экстрагировали этилацетатом и высушивали. После выпаривания растворителя остаток хроматографировали на силикагеле (с элюированием 75% этилацетата в гексане), что дало указанное в заголовке соединение в виде пены (80,5 мг, выход 80%).Getting 13. 10-Deoxytaxoter

10-Desacetoxy-7-O-triethylsilylbaccatin III (100 mg, 0.156 mmol) was placed in a flask under argon and dissolved in dry tetrahydrofuran (1.5 ml). After cooling to -40 ^° C, n-butyllithium (1.45 M in hexanes, 0.119 ml, 0.170 mmol) was added dropwise, followed by (3R, 4S) -1-tert-butoxycarbonyl-4-phenyl-3-triethylsilyloxy- 2-azetidinone (94.2 mg, 0.25 mmol) in tetrahydrofuran (0.5 ml) for 2 minutes. The mixture was immediately heated to 0 ^° C. and stirred for 45 minutes before being quenched with saturated ammonium chloride (3 ml). The mixture was extracted with ethyl acetate, dried and concentrated. Silica gel chromatography (eluting with 30% ethyl acetate in hexane) gave 10-deoxy-2 ', 7-bis-O- (triethylsilyl) taxotere as a foam (125 mg, 76% yield). This compound (100 mg, 0.098 mmol) was immediately dissolved in acetonitrile (2 ml) at -5 ^° C and treated with hydrochloric acid (0.037 ml, 36%, 12 M). The mixture was stirred for 2 hours at -5 ^° C, after which it was sharply cooled with aqueous bicarbonate, extracted with ethyl acetate and dried. After evaporation of the solvent, the residue was chromatographed on silica gel (eluting with 75% ethyl acetate in hexane) to give the title compound as a foam (80.5 mg, 80% yield).

 The general procedure carried out in Preparation 13 can be adapted to produce other compounds of formula (Ia) with the starting components baccatin III and azetidinone; examples of other compounds of formula (Ia) are listed in table IV. It is understood that although the following compounds are shown with free hydroxy groups, with appropriate selection of different hydroxy protecting groups, any of the protecting groups at position 2 ', 7 or 10 can be selectively removed without affecting other available protecting groups.

Getting 14 Vis (methylthiomethyl) ether
SH ₃ SCH ₂ OCH ₂ SCH ₃
To a solution of 1,1'-dichlorodimethyl ether (3.0 g, 26.3 mmol) in acetone (100 ml) at 0 ^° C was added sodium iodide (8.23 g, 55.23 mmol), and the mixture was stirred at this temperature 20 minutes Then sodium thiomethoxide (1.84 g, 5.23 mmol) was added in four portions, and the resulting solution was stirred for another 1 hour. After that, the heterogeneous solution was filtered through a celite pad, and the filtrate was concentrated in vacuo. The residual oil was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous layer was removed and then extracted with ethyl acetate, then the combined organics were treated with a mixture of saturated aqueous sodium bicarbonate and 5% aqueous sodium thiosulfate with a ratio of 1: 1 (v / v). Then the organics were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residual oil was purified by flash chromatography (30: 1, hexanes: ethyl acetate) to obtain 1.9 g of a yellow oil, which was then distilled using a Kugelor apparatus (120-130 ^° C, 20 mmHg), which gave 1 5 g (45%) of the title compound as a colorless oil:
¹ H NMR (300 MHz, CDCl ₃ ): δ 4.73 (4H, s), 2.15 (6H, s),
Getting 15. Dibenzylmethylthiomethylphosphate
CH ₃ SCH ₂ OP (O) (OBu) ₂
To a solution of bis (methylthiomethyl) ether (30 mg, 2.34 mmol) and molecular sieves (300 mg) in tetrahydrofuran (THF) (100 ml), dibenzyl phosphate (2.74 g, 9.85 mmol) was added at room temperature and then N-iodosuccinimide (608 mg, 2.71 mmol) and the solution was stirred for 4 hours. The reaction mixture was then diluted with ethyl acetate and filtered through a pad of celite. The filtrate was treated with a mixture of a saturated aqueous solution of sodium bicarbonate and a 5% aqueous solution of sodium thiosulfate with a ratio of 1: 1 (v / v). The colorless organic extract was then washed with brine, dried over sodium sulfate and concentrated in vacuo to give 600 mg (69%) of the title compound:
¹ H NMR (300 MHz, CDCl ₃ ): δ 7.35 (10H, s), 5.29 (2H, d, J = 12.2 Hz), 5.08 (4H, dd, J = 8.0 and 1.0 Hz), 4.68 (2H, s), 2.10 (3H, s).

Examples
The following examples are provided to illustrate the synthesis of the present compounds in accordance with the present invention and should not be construed as limiting the scope of the invention in any way. Specialists in the art can adapt these methods (without unnecessary experiments) to the synthesis of compounds, although not specifically disclosed, but are included in the scope of the present invention.

К тщательно перемешанной смеси паклитаксела (0,85 г, 1 ммоль) и диметилсульфида (0,72 мл, 8 ммоль) в сухом ацетонитриле (10 мл) при 0^oC добавляли бензоилпероксид (0,98 г, 4 ммоль). Перемешивание продолжали в течение 2,5 часов при 0^oC. Ход реакции контролировали путем тонкослойной хроматографии на силикагеле в системе растворителей толуол:кетон (2:1, объем/объем) (R_fтакс.= 0,38, R_fпрод.= 0,64), и когда видели образование продуктов более высокой полярности, то реакцию быстро прекращали путем выпаривания растворителей с использованием роторного испарителя Ротавапор при 30^oC. Анализ путем тонкослойной хроматографии показал наличие некоторых количеств непрореагировавших паклитаксела и 2',7-O-бис(метилтиометил) паклитаксела. Отделение указанного в заголовке соединения от реакционной смеси производили путем колоночной флеш-хроматографии на силикагеле 60 (40 - 63 мкм). EM Sciense (100 мл), диаметр колонки 2 дюйма (50,6 мм) с использованием системы растворителей этилацетат-гексан (1: 1, объем/объем) (R_fпрод.=0,34). Продукт (552 мг, выход 60%) извлекали из фракций 12 - 18 (каждая фракция приблизительно 20 мл).Example 1. 7-O-Phosphonoxymethylpaclitaxel and its monosodium salt
(a) Preparation of 7-O-methylthiomethylpaclitaxel

To a thoroughly mixed mixture of paclitaxel (0.85 g, 1 mmol) and dimethyl sulfide (0.72 ml, 8 mmol) in dry acetonitrile (10 ml) was added benzoyl peroxide (0.98 g, 4 mmol) at 0 ^° C. Stirring was continued for 2.5 hours at 0 ^° C. The progress of the reaction was monitored by thin layer chromatography on silica gel in a toluene: ketone (2: 1, vol / vol) solvent system (R _ftax. = 0.38, R _fprod. = 0 , 64), and when the formation of products of higher polarity was observed, the reaction was quickly stopped by evaporation of the solvents using a Rotavapor rotary evaporator at 30 ^° C. Analysis by thin-layer chromatography showed the presence of some amounts of unreacted paclitaxel and 2 ', 7-O-bis ( methylthiomethyl) paclitaxel. The title compound was separated from the reaction mixture by flash column chromatography on silica gel 60 (40 - 63 μm). EM Sciense (100 ml), 2-inch column diameter (50.6 mm) using an ethyl acetate-hexane (1: 1, v / v) solvent system (R _fprod. = 0.34). The product (552 mg, 60% yield) was recovered from fractions 12-18 (each fraction 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 (MeOH): λ _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 .

¹ H-NMR (CDCl ₃ ), δ: 1.15 (3H, s), 1.19 (3H, s), 1.73 (3H, s), 1.79 (H, s), 1.90 (3H, d), 2.09 (3H, s), 2.16 (3H, s), 2.29 (2H, d), 2.35 (3H, s), 2.77 (H, m) , 3.70 (H, d), 3.83 (H, d), 4.17 (H, d), 4.26 (H, d, overlaps with H, d), 4.63 (2H, t ), 4.77 (H, dd), 4.91 (H, d), 5.65 (H, d), 5.77 (H, dd), 6.16 (H, dd), 6.48 (H, s), 7.07 (H, d), 7.29 - 7.50 (10H, m), 7.57 (H, m), 7.73 (2H, d), 8.08 ( 2H, d).

К смеси 7-O-метилтиометилпаклитаксела (119 мг, 0,13 мМ) и порошкообразных молекулярных сит

(приблизительно 120 мг) в сухом 1,2-дихлорметане (5 мл) добавляли раствор N-иодсукцинимида (45 мг, 0,2 мМ) и дибензилфосфата (55 мг, 0,2 мМ) в сухом тетрагидрофуране (4 мл). Реакционную смесь перемешивали при комнатной температуре 16 часов. Ход реакции контролировали тонкослойной хроматографией в системе толуол: ацетон (2: 1, объем/объем) (R_fпрод.= 0,48). Молекулярные сита удаляли путем фильтрования через Целит 545, и фильтрат экстрагировали метиленхлоридом (100 мл). Органический слой промывали 1%-ным раствором тиосульфата натрия (примерно 100 мл) и 0,5 М бикарбоната натрия (100 мл) и рассолом. Экстракт фильтровали через сепаратор Ватмана для разделения фаз, и выпаривали растворители. Очистка на флеш-колонке с силикагелем 60 в смеси метиленхлорид:этилацетат (2:1, объем/объем) дала 7-O-дибензилфосфонооксиметилпаклитаксел (41,5 мг).(b) Preparation of 7-O-dibenzylphosphonooxymethylpaclitaxel

To a mixture of 7-O-methylthiomethylpaclitaxel (119 mg, 0.13 mmol) and powdered molecular sieves

(approximately 120 mg) in dry 1,2-dichloromethane (5 ml) was added a solution of N-iodosuccinimide (45 mg, 0.2 mmol) and dibenzyl phosphate (55 mg, 0.2 mmol) in dry tetrahydrofuran (4 ml). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by thin layer chromatography in a toluene: acetone system (2: 1, v / v) (R _fprod. = 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 was washed with a 1% solution of sodium thiosulfate (approximately 100 ml) and 0.5 M sodium bicarbonate (100 ml) and brine. The extract was filtered through Whatman separator to separate the phases, and the solvents were evaporated. Purification on a flash column with silica gel 60 in a mixture of methylene chloride: ethyl acetate (2: 1, v / v) gave 7-O-dibenzylphosphonooxymethyl paclitaxel (41.5 mg).

7-O-Дибензилфосфонооксиметилпаклитаксел (41,5 мг) растворяли в этилацетате (5 мл) и добавляли 10%-ный палладий на древесном угле (20 мг). Осуществляли гидрогенизацию при 40 фунтах на кв. дюйм (275 кПа) при комнатной температуре в течение 1 часа. Ход реакции контролировали тонкослойной хроматографией в системе хлороформ:метанол:вода (120:45:8, объем/объем). Очистка путем препаративной тонкослойной хроматографии (несущая силикагель пластика 20х20х0,05 см в аналитической системе) дала 7-O-фосфонооксиметилпаклитаксел (26 мг, выход 75%).(c) Preparation of 7-O-phosphonooxymethylpaclitaxel and its Monosodium Salt

7-O-Dibenzylphosphonooxymethylpaclitaxel (41.5 mg) was dissolved in ethyl acetate (5 ml) and 10% palladium on charcoal (20 mg) was added. Hydrogenation was carried out at 40 psi. inch (275 kPa) at room temperature for 1 hour. The reaction progress was monitored by thin layer chromatography in a chloroform: methanol: water system (120: 45: 8, v / v). Purification by preparative thin layer chromatography (silica gel-containing plastic 20x20x0.05 cm in the analytical system) gave 7-O-phosphonooxymethylpaclitaxel (26 mg, 75% yield).

 Since decomposition of 7-O-dibenzylphosphonooxymethylpaclitaxel was observed during silica gel purification, the hydrogenation procedure was modified. Thus, the crude extract of 7-O-dibenzylphosphonooxymethylpaclitaxel was hydrogenated without any purification. Hydrogenation of the crude 7-O-dibenzylphosphonooxymethylpaclitaxel extract was carried out at 60 psi. inch (400 kPa) for 24 hours.

7-O-phosphonooxymethylpaclitaxel (70 mg) was dissolved in 5 ml of acetone-water solution (1: 1) and diluted with water to 50 ml. Dry sodium bicarbonate (18 mg, 1.2 equiv.) Was added. Acetone was evaporated at room temperature using a rotary evaporator and the remaining aqueous solution was lyophilized. The crude 7-O-phosphonooxymethylpaclitaxel monosodium salt is purified by chromatography on a reverse phase C18 column in a water: acetonitrile system (70: 30, v / v). The eluate was monitored by analytical HPLC / 15 cm, Jones C18 column, 1 ml / min, l = 230/270 nm / in acetonitrile buffer: 0.05 M ammonium acetate (45:55, v / v), pH = 7, R _t = 2.09 min. The fractions containing the desired product were combined, acetonitrile was evaporated, and the remaining aqueous solution was lyophilized, resulting in a monosodium salt (112 mg) of 7-O-phosphonoxymethylpaclitaxel.

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

UV spectrum (MeOH): λ _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 .

¹ H-NMR (acetone-d ₆ / D ₂ O), δ: 8.05 (2H, d), 7.92 (2H, d), 7.65 (1H, dd), 7.58-7. 35 (9H, m, overlap), 7.23 (1H, dd), 6.38 (1H, s), 6.08 (1H, t), 5.65 (1H, d), 5.60 (1H d), 5.10 (1H, bs), 4.99 (1H, ds), 4.97 (1H, bs), 4.80 (1H, d), 4.28 (1H, dd), 4 11 (2H, s), 3.79 (1H, d), 2.94 (1H, m), 2.35 (3H, s), 2.35 - 2.10 (1H, m), 2, 13 (3H, s), 1.95 (3H, s), 1.84 (1H, m), 1.67 (3H, s), 1.13 (6H, s, overlap).

К перемешанному раствору паклитаксела (150 мг, 0,176 ммоль) и N,N-диизопропилэтиламина (93 мкл, 0,534 ммоль, 3 эквив.) в безводном метиленхлориде (4 мл) при комнатной температуре добавляли бензилхлорформиат (75 мкл, 0,525 ммоль, 3 эквив.). Реакционную смесь перемешивали при комнатной температуре 3 часа, концентрировали до 32 мл и очищали на колонке с силикагелем с использованием в качестве элюента смеси (1:1) этилацетат:гексаны, в результате чего получали указанное соединение в виде белого порошка (150 мг, выход 86%); т.пл. 140 - 150^oC (разложение).Example 2. An alternative method of obtaining 7-O-phosphonooxymethylpaclitaxel
(a) Preparation of 2'-O- (benzyloxycarbonyl) paclitaxel

To a mixed 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) was added benzyl chloroformate (75 μl, 0.525 mmol, 3 equiv.) At room temperature. ) The reaction mixture was stirred at room temperature for 3 hours, concentrated to 32 ml and purified on a silica gel column using ethyl acetate: hexanes (1: 1) as an eluent to give the title compound as a white powder (150 mg, yield 86 %); so pl. 140 - 150 ^o C (decomposition).

К охлажденному (сухой лед-CCl₄, температура ванны -30^oC) раствору 2'-O-(бензилоксикарбонил)паклитаксела (4,395 г, 5,0 ммоль) в сухом ацетонитриле (80 мл) добавляли последовательно диметилсульфид (3,6 мл, 40 ммоль) и бензоилпероксид (4,9 г, 20,247 ммоль). Через 10 минут, при -30^oC, убирали охлажденную ванну и тщательно перемешивали реакционную смесь в течение 2 ч при комнатной температуре. Затем реакционную смесь разбавляли этилацетатом до объема 200 мл и промывали водой и рассолом. Органический слой высушивали (MgSO₄), после чего выпаривали растворитель, получив в результате остаток, который держали под вакуумом 18 ч для удаления диметилсульфоксида, который присутствовал как побочный продукт реакции. Остаток очищали на колонке с силикагелем с использованием в качестве элюента сначала смеси (1:2) этилацетат-гексан для удаления менее полярных примесей, а затем смеси (1:1) этилацетат-гексан, что дало в результате ожидаемое указанное в заголовке соединение в виде пены. Тщательно его растирали с сухим диэтиловым эфиром и фильтровали, получив указанное в заголовке соединение в виде рыхлого твердого вещества (5,0 г, 95%); т.пл. 120 - 122^oC.(b) Preparation of 2'-O- (benzyloxycarbonyl) -7-O-methylthiomethylpaclitaxel

To a cooled (dry ice-CCl ₄ bath temperature -30 ^° C.) solution of 2'-O- (benzyloxycarbonyl) paclitaxel (4.395 g, 5.0 mmol) in dry acetonitrile (80 ml) was added successively dimethyl sulfide (3.6 ml , 40 mmol) and benzoyl peroxide (4.9 g, 20.247 mmol). After 10 minutes, at -30 ^° C, the cooled bath was removed and the reaction mixture was thoroughly stirred for 2 hours 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 (MgSO ₄ ), after which the solvent was evaporated, yielding a residue, which was kept under vacuum for 18 h to remove dimethyl sulfoxide, which was present as a reaction by-product. The residue was purified on a silica gel column using first (1: 2) ethyl acetate-hexane as an eluent to remove less polar impurities, and then a mixture (1: 1) ethyl acetate-hexane, which gave the expected title compound as foam. It was triturated thoroughly with dry diethyl ether and filtered to give the title compound as a friable solid (5.0 g, 95%); so pl. 120 - 122 ^o C.

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 (CDCl ₃ ), δ: 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, s), 6.920 (H, d, NH), 7.322-8.22 (15H , m).

 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 dimethyl sulfoxide (10 ml) was added dropwise acetic anhydride (10 ml). The resulting mixture was stirred at room temperature for 18 hours under N ₂ , diluted with ethyl acetate (100 ml) and washed thoroughly with cold 6% sodium bicarbonate solution (6 x 30 ml), chloride water (6 x 30 ml) and brine. The (organic layer) MgSO _{4 was} dried, and the solvent was evaporated to give a residue. It was purified on a silica gel column and eluted with methylene chloride, 5% acetonitrile methylene chloride and 10% acetonitrile methylene chloride, to give the expected title compound (1.86 g, 87.7%). This compound is identical to that obtained by the previously described method using dimethyl sulfide and benzoyl peroxide.

К раствору 2'-O-(бензилоксикарбонил)-7-O- метилтиометилпаклитаксела (5,0 г, 5,5396 ммоль) в сухом 1,2-дихлорэтане (120 мл) добавляли активированные порошкообразные молекулярные сита

(5,0 г). К этой смеси добавляли по каплям при комнатной температуре раствор смеси N-иодсукцинимида (1,61 г, 7,1632 ммоль) и дибензилфосфата (1,97 г, 7,1632 ммоль) в сухом тетрагидрофуране (90 мл). После интенсивного перемешивания при комнатной температуре в течение 30 мин реакционную смесь фильтровали через целит, и фильтрат упаривали досуха, получив красный остаток. Остаток растворяли в этилацетате (100 мл), промывали холодным 6%-ным раствором NaHSO₃ (2 х 50 мл), холодным 6%-ным раствором NaHCO₃ (2 х 50 мл) и рассолом (1 х 50 мл). Органический слой высушивали (MgSO₄), и выпаривали растворитель, получив в результате твердую массу, которую растирали в порошок с сухим диэтиловым эфиром и фильтровали, получив указанное в заголовке соединение в виде цвета слоновой кости твердого вещества (5,9 г, 97%); т.пл. 124 - 127^oC.(c) Preparation of 2'-O- (bnzyloxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel

To a solution of 2'-O- (benzyloxycarbonyl) -7-O-methylthiomethylpaclitaxel (5.0 g, 5.5396 mmol) in dry 1,2-dichloroethane (120 ml) was added activated powder 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 dibenzyl phosphate (1.97 g, 7.1632 mmol) in dry tetrahydrofuran (90 ml). After vigorous stirring at room temperature for 30 minutes, the reaction mixture was filtered through celite, and the filtrate was evaporated to dryness to give a red residue. The residue was dissolved in ethyl acetate (100 ml), washed with cold 6% NaHSO ₃ solution (2 x 50 ml), cold 6% NaHCO ₃ solution (2 x 50 ml) and brine (1 x 50 ml). The organic layer was dried (MgSO ₄ ), and the solvent was evaporated, yielding a solid mass, which was triturated with dry diethyl ether and filtered to obtain the title compound as an ivory solid (5.9 g, 97%) ; so pl. 124 - 127 ^o C.

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 (CDCl ₃ ), δ: 1,160 (3H, s), 1,703 (3H, s), 1,985 (3H, s), 2,164 (3H, s), 2,420 (3H, s), 3,854 (H, d), 4.151 (H, d), 4.216 (H, m), 4.298 (H, d), 4.873 (H, d), 5.043 (6H, m), 5.140 (2H, d), 5.417 (H, d), 5.670 (H, d), 5.971 (H, dd), 6.241 (H, t), 6.317 (H, s), 6.912 (H, d, NH), 7.280 - 8.115 (25H, m).

(d) Preparation of 7-O-phosphonooxymethylpaclitaxel
To a solution of 2'-O- (benzyloxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (6.0 g, 4.7095 mmol) in ethyl acetate (120 ml) was added 10% Pd / C (6.0 g), and the mixture was hydrogenated at 60 psi inch (400 kPa) for 24 hours. The reaction mixture was filtered through celite and, evaporating the solvent, 4.07 g of a crude residue was obtained. It was purified on a short silica gel column by eluting successively with a mixture of chloroform with 10%, 20% and 40% methanol to give the title compound as a white solid (3.2 g, 71%); t. pl. 155 - 158 ^o C.

This product had the same R _f (thin layer chromatography) and the same retention time (HPLC - high pressure liquid chromatography) as an 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): λ _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 .

¹ NMR (acetone-d ₆ / D ₂ O), δ: 1.08 (3H, s), 1.10 (3H, s), 1.63 (3H, s), 1.88 (3H, s) , 1.96 (H, m), 2.13 (3H, s), 2.32 (3H, s), 2.89 (H, m), 3.76 (H, d), 4.19 ( H, m), 4.89 (H, dd), 5.09 (H, dd), 5.55 - 5.60 (2H, overlapping doublets), 6.04 (H, t), 6.32 ( H, s), 7.20 (H, t), 7.34 - 7.67 (10H, overlapping multiplets), 7.87 (2H, dd), 8.02 (2H, dd).

К раствору паклитаксела (4,35 г, 5,1 ммоль) в сухом метиленхлориде (51 мл) добавляли N,N-диизопропилэтиламин (2,67 мл, 15,3 ммоль) и затем этилхлорформиат (1,46 мл, 15,3 ммоль). Реакционную смесь перемешивали при 0^oC 2 часа, а затем при комнатной температуре еще 1 час. Реакционную смесь разбавляли этилацетатом (400 мл), органическую фазу промывали насыщенным раствором NaHCO₃ (2 х 30 мл) и рассолом (30 мл). Полученную органическую фазу высушивали над MgSO₄ и получили неочищенное указанное в заголовке соединение (93%), которое использовали на следующей стадии без дополнительной очистки.Example 3. 2'-O- (Ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel
(a) Preparation of 2'-O- (ethoxycarbonyl) paclitaxel

To a solution of paclitaxel (4.35 g, 5.1 mmol) in dry methylene chloride (51 ml) was added N, N-diisopropylethylamine (2.67 ml, 15.3 mmol) and then ethyl chloroformate (1.46 ml, 15.3 mmol). The reaction mixture was stirred at 0 ^° C. for 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 NaHCO ₃ solution (2 x 30 ml) and brine (30 ml). The resulting organic phase was dried over MgSO _{4 to} give the crude title compound (93%), which was used in the next step without further purification.

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

High-resolution mass spectrum (FAB / NOBA, external CSI / Gli standard): [M + H] ⁺ , m / z 926.3588 observed, C ₅₀ H ₅₆ NO ₁₆ , calculated value: 926.3599 (deviation Δ = 1 , 2 ppm).

¹ H-NMR (CDCl ₃ ), δ: 1.13 (3H, s), 1.23 (3H, s), 1.30 (3H, t), 1.67 (3H, s), 1.92 (3H, s), 2.21 (3H, s), 2.37 (H, d), 2.45 (3H, s), 2.54 (H, m), 3.80 (H, d) , 4.15 - 4.32 (4H, overlapping multiplets), 4.43 (H, dd), 4.96 (H, d), 5.42 (H, d), 5.68 (H, d) 5.98 (H, dd), 6.28 (2H, overlapping multiplets), 7.00 (H, d), 7.34 - 7.59 (11H, overlapping multiplets), 7.74 (2H, d ), 8.12 (2H, d).

Alternative procedure
Paclitaxel (5.40 g, 6.324 mmol) in dry dichloromethane (63 ml) was cooled to 0 ^° C. and treated with undiluted N, N-diisopropylethylamine (3.30 ml, 3 equiv.) And then with undiluted ethyl chloroformate (1.81 ml, 3 equiv.) Dropwise over 5 minutes. The reaction was monitored by thin layer chromatography (50% ethyl acetate in hexane). After 2 hours at 0 ^° C. and 16 hours 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 (MgSO ₄ ) and evaporation gave the crude title compound which was purified by precipitation: dichloromethane (about 100 ml) was added, then it was cooled and hexane (about 60 ml) was added to the cloud point. After cooling in ice for several hours, a solid was collected by filtration (5.17 g, 88% yield).

Alternative procedure
In a 3 L single-necked flask dried above the flame, paclitaxel (99.0 g, 115.9 mmol) was dissolved in 1350 ml of dry methylene chloride in argon. The solution was cooled to -10 ^° C. N, N-diisopropylethylamine (52.4 g, 405.7 mmol) was added slowly (approximately 3 min), and then ClCO ₂ Et (31.45 g, 289.8 mmol; time was added). add about 15 minutes). The resulting mixture was stirred overnight (16 hours) at -4 ^° C. Thin layer chromatography showed that the reaction was not completed. Therefore, another weighed portion of N, N-diisopropylethylamine (2.62 g, 20.28 mmol) was added and then ClCO ₂ Et (2.20 g, 20.28 mmol) and stirring was continued for 3 hours at -4 ^° C. Thin layer chromatography showed lack of source material. The cold mixture was diluted with ethyl acetate (1.5 L) and transferred to a separatory funnel. Then it was washed with 5% KHSO ₄ solution (2 x 500 ml), water (1 x 500 ml), 5% KHSO ₄ solution (1 x 500 ml), water (1 x 500 ml), saturated NaHCO solution ₃ (2 x 500 ml) and brine (2 x 500 ml), dried (MgSO ₄ ) and, removing the solvent in vacuo, 147 g of crude product was obtained. The residue was dissolved in hot methylene chloride (800 ml, bath temperature 42 ^° C.), and hexanes (530 ml) were added dropwise with stirring while maintaining the indicated temperature. The crystallizing mixture was set aside for 3 hours at room temperature, and then put in a cold room (0 ^o C) overnight. Heavy white crystals were collected by filtration and washed with a mixture of hexanes - CH ₂ Cl ₂ (1: 1, v / v) (2 x 200 ml). After drying on a vacuum filter for 1 hour, they were dried in vacuo (approximately 1.0 mmHg) overnight to give 95.7 g (89% yield) of the title compound (uniformity index measured by HPLC = 98.5%).

К раствору 2'-O-(этоксикарбонил)паклитаксела (4,38 г, 4,7 ммоль) в сухом диметилсульфоксиде (12,5 мл) добавляли уксусный ангидрид (12,5 мл). Реакционную смесь перемешивали 24 часа при комнатной температуре и затем разбавляли этилацетатом (500 мл), промывали насыщенным раствором NaHCO₃ (3 х 40 мл) и водой (2 х 40 мл). Полученный органический слой высушивали над MgSO₄, и выпаривали растворители в вакууме досуха. Остаток очищали путем хроматографии на силикагеле (40% этилацетата в гексанах) и получили в результате требуемое указанное в заголовке соединение (4,39 г, 94%).(b) Preparation of 2'-O- (ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel

To a solution of 2'-O- (ethoxycarbonyl) paclitaxel (4.38 g, 4.7 mmol) in dry dimethyl sulfoxide (12.5 ml) was added acetic anhydride (12.5 ml). The reaction mixture was stirred 24 hours at room temperature and then diluted with ethyl acetate (500 ml), washed with saturated NaHCO ₃ solution (3 x 40 ml) and water (2 x 40 ml). The resulting organic layer was dried over MgSO ₄ , and the solvents were evaporated to dryness in vacuo. The residue was purified by silica gel chromatography (40% ethyl acetate in hexanes) to give the desired title compound (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 CSI / Gly reference): [M + H] ⁺ , m / z 986.3646 (calculated value: 986.3633, deviation Δ = 1.3 ppm).

¹ H NMR (CDCl ₃ ): δ: 1.18 (3H, s), 1.20 (3H, s), 1.30 (3H, s), 1.75 (3H, s), 1.84 (H, m), 2.09 (3H, s), 2.11 (3H, s), 2.16 (3H, s), 2.24 (H, d), 2.37 (H, d) 2.45 (3H, s), 2.80 (H, m), 3.68 (H, d), 4.08 - 4.33 (5H, m, overlap), 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, s), 7.00 (H, d), 7.32 - 7.61 (11H, m, overlap), 7.73 (2H, dd), 8.11 (2H, dd).

Alternative procedure
2'-O- (Ethoxycarbonyl (paclitaxel (2.260 g, 2.4406 mmol) was dissolved in anhydrous dimethyl sulfoxide (6 ml) and acetic anhydride (6 ml) was added in one portion at room temperature. The reaction was monitored by HPLC (C18 analytical column; 60% acetonitrile - 40% 10 mM ammonium phosphate buffer, pH 6) After 30 hours, the solution was diluted with ethyl acetate (250 ml) and washed with saturated aqueous bicarbonate (3 times) and then with water and brine. After drying over magnesium sulfate and filtering, the crude product chromatographed on silica gel (40% ethyl acetate in hex not) and the title compound was obtained as a white foam (2.030 g, 91%) with a purity of 90% as determined by HPLC (high pressure liquid chromatography). Part of the product was further purified on a second column (5% acetonitrile in dichloromethane), yielding as a result, a material having a purity of approximately 97% as determined by HPLC.

An alternative way to obtain 2'-O- (ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel
2'-O- (ethoxycarbonyl) paclitaxel (4.170 g, 4.503 mmol) was dissolved in anhydrous acetonitrile (68 ml) at -40 ^° C, and dimethyl sulfide (3.2 ml, 44.10 mmol) was added, followed by benzoyl peroxide (4.400 g, 18.24 mmol). The mixture was placed in an ice bath and stirred at 0 ^° C., and the progress of the reaction was monitored by thin layer chromatography (40% ethyl acetate in hexane). After 3 hours, without detecting the starting material, the solution was treated by adding ethyl acetate (250 ml) and saturated aqueous sodium bicarbonate (100 ml). Then the organic phase was washed with bicarbonate, water and brine, after which it was dried over magnesium sulfate and filtered. The residue was purified by flash chromatography on silica gel (4% acetonitrile in dichloromethane) to give the title compound as a white foam (2.571 g, 58% yield). The purity of this sample was evaluated by HPLC and it was more than 97%. NMR spectrum was identical to that presented above.

Alternative Procedure for the Preparation of 2'-O- (ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel
2'-O- (Ethoxycarbonyl) paclitaxel (49.3 g, 53.2 mmol) was placed in a 1 L flask dried over a flame and dissolved in dry acetonitrile (500 ml) at room temperature. Methyl sulfide (39.1 ml, 0.532 mmol) was quickly added via syringe. The stirred reaction mixture was cooled to -16 ^° C in a salt and ice bath, and solid benzoyl peroxide (51.6 g, 0.213 mmol) was added to the mixture in one portion. (For the full implementation of the reaction, the full four equivalents are required) Stirring was continued for 30 minutes, during which the temperature rose to about -10 ^° C. Throughout this period, the reaction medium remained heterogeneous (benzoyl peroxide did not completely dissolve). The cooling bath was replaced with ice water, as a result of which the temperature rose to 0 ^° C, and the remaining benzoyl peroxide dissolved in about 5 minutes after the temperature was raised. After stirring for another 2.5 hours at 0 ^° C., the reaction, as thin-layer chromatography showed, was completed. By removing the solution on a rotary evaporator, the solution volume was reduced to about 200 ml, after which the solution was transferred to a separatory 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) of saturated NaHCO ₃ and 5% K ₂ CO ₃ (v / v) (2 x 500 ml), saturated NaHCO ₃ (2 x 500 ml), semi-saturated brine (1 x 500 ml) and brine (1 x 500 ml), dried (MgSO ₄ ) and, removing the solvent in vacuo, 67.0 g of crude product was obtained. It was dissolved in acetone (200 ml), heated to 40 ^° C. in a water bath, after which hexanes were added dropwise with stirring until a turbidity (400 ml) was noticed. The crystallizing mixture was left to stand for 3 hours at room temperature, and then transferred to a cold chamber (0 ^° C.), where it was kept overnight (16 hours). A thick layer of caked sediment formed. The solid was collected by filtration and washed with a mixture (3: 1, v / v) hexanes-acetone (2 x 50 ml). The resulting white crystals were dried on a vacuum filter for 1 hour and then in vacuum (about 0.5 mmHg) all night, and as a result, 47.5 g (91% yield) of the title compound (uniformity index at measured by HPLC was 94.8%).

К смеси 2'-O-(этоксикарбонил)-7-O-метилтиометилпаклитаксела (5,677 г, 5,76 ммоль) и молекулярных сит

(5,7 г) в метиленхлориде (100 мл) при комнатной температуре добавляли раствор N-иодсукцинимида (1,953 г, 8,65 ммоль) и дибензилфосфата (2,41 г, 8,65 ммоль) в тетрагидрофуране. Реакционную смесь перемешивали 40 мин при комнатной температуре. После этого периода реакция, как показала тонкослойная хроматография, была завершена. Реакционную смесь фильтровали через целит, и фильтрат концентрировали в вакууме, получив в результате буроватый остаток, который разбавляли этилацетатом (800 мл), органическую фазу промывали 1%-ным Na₂SO₃ (2 x 80 мл), а затем 5%-ным рассолом (2 x 50 мл). Органическую фазу концентрировали в вакууме и высушивали. Хроматография полученного остатка (50 - 60% этилацетата в гексанах) дала целевое указанное в заголовке соединение (6,23 г, 89%).(c) Preparation of 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel

To a mixture of 2'-O- (ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel (5.677 g, 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 dibenzyl phosphate (2.41 g, 8.65 mmol) in tetrahydrofuran. The reaction mixture was stirred for 40 minutes at room temperature. After this period, the reaction, as thin layer chromatography showed, 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 was washed with 1% Na ₂ SO ₃ (2 x 80 ml), and then 5% brine (2 x 50 ml). The organic phase was concentrated in vacuo and dried. Chromatography of the obtained residue (50-60% ethyl acetate in hexanes) gave the title compound (6.23 g, 89%).

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

High-resolution mass spectrum (FAB / NOBA, external CSI / gli standard): [M + Na] ⁺ , m / z 1216, 4291
(C ₆₅ H ₇₁ NO ₂₀ P, calculated value: 1216, 4307, deviation Δ = 1.3 ppm).

¹ H NMR (CDCl ₃ ), δ: 1.18 (3H, s), 1.21 (3H, s), 1.30 (3H, t), 1.67 (6H, s), 1.80 ( H, s), 1.93 (H, m), 1.99 (3H, d), 2.18 (3H, s), 2.23 (H, m), 2.38 (H, m), 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 - 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 - 7.61 (11H, overlapping multiplets), 7.75 (2H, dd), 8.12 (2H, dd).

Alternative procedure
To a solution of 2'-O- (Ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel (350 mg, 0.355 mmol) in anhydrous tetrahydrofuran (8 ml) was added a solution of N-iodosuccinimide (120 mg, 0.532 mmol) and dibenzyl phosphate (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 completed, as indicated by less than 5% of the detected starting material. The solution was diluted with ethyl acetate (75 ml) and washed with a 1% aqueous solution of bisulfite (2 x 50 ml) and brine (50 ml). After quick drying over magnesium sulfate and filtration, the solvent was evaporated. Silica gel flash chromatography (45% ethyl acetate in hexane) gave the title compound as a white foam (281 mg, 65%). HPLC analysis showed a purity of approximately 95%.

помещали в высушенную над пламенем одногорлую колбу емкостью 1 л, которую затем соединяли с вакуумной линией (примерно 0,5 мм рт.ст.). Сита нагревали с помощью прибора для сушки горячим воздухом в течение примерно 10 мин с встряхиванием вручную. После охлаждения под вакуумом в колбу вводили аргон и добавляли 2'-O-(Этоксикарбонил)-7-O-метилтиометилпаклитаксел (37,5 г, 38,03 ммоль), а затем дибензилфосфат (14,3 г, 53,24 ммоль) и тетрагидрофуран (400 мл). Гетерогенную смесь интенсивно перемешивали 15 минут при комнатной температуре посредством магнитной мешалки. В отдельной высушенной над пламенем колбе растворяли N-иодсукцинимид (10,7 г, 47,54 ммоль) в тетрагидрофуране (50 мл) под аргоном. (Во время приготовления раствора NIS переноса жидкости и хода реакции сосуды покрывали алюминиевой фольгой для защиты от света). Затем к реакционной смеси медленно (10 мин) добавляли посредством шприца приготовленный раствор. Колбу, содержащую NIS, промывали 5 мл ТГФ и переносили в реакционную смесь, которую затем перемешивали 2 часа при комнатной температуре. Анализ путем тонкослойной хроматографии показал отсутствие исходного материала. Раствор темно-красного цвета фильтровали через слой целита непосредственно в интенсивно перемешиваемую двухфазную смесь, содержащую этилацетат (500 мл) 10%-ный водный раствор тиосульфата натрия (300 мл) и насыщенный раствор бикарбоната натрия (200 мл). Через несколько секунд красный цвет исчез и раствор стал бесцветным. Слой целита промывали этилацетатом (примерно 100 мл) и оба слоя жидкости переносили в делительную воронку. Органический слой разбавляли 1 литром этилацетата, разделяли слои, и органический слой промывали смесью насыщенного NaHCO₃ и 5%-ного K₂CO₃, а затем насыщенным NaHCO₃ (2 x 500 мл), полунасыщенным рассолом (1 x 500 мл) и рассолом (1 x 500 мл). Экстракт высушивали безводным MgSO₄ и фильтровали. Затем его обрабатывали 5,0 г нейтрального норита (древесный уголь) путем перемешивания при комнатной температуре в течение 15 минут. Опять фильтровали его через слой целита, и, удалив растворитель при пониженном давлении, получили 52 г неочищенного продукта. Его растворяли в смеси толуола и метиленхлорида (280 мл/25 мл), и к полученному раствору добавляли по каплям гексаны (20 мл). После 3 часов выдерживания при комнатной температуре кристаллизующуюся смесь всю ночь держали при 0^oC. На стенках колбы образовалось твердое вещество бледно-желтого цвета. После декантирования маточного раствора остаток тритурировали с толуолом (50 мл), фильтровали, промывали толуолом и высушивали на вакуумном фильтре в течение 30 минут. Затем переносили его в эксикатор с драйеритом, после чего высушивали в вакууме (примерно 0,5 мм рт.ст.) в течение 4 часов, получив в результате 24,4 г (выход 53%) указанного в заголовке соединения (показатель гомогенности при определении путем HPLC был равен 95,9%). Маточный раствор упаривали досуха, тритурировали с тоуолом (100 мл), фильтровали, промывали толуолом и высушивали на вакуумном фильтре в течение 30 минут. После высушивания в эксикаторе, как описано выше, получили 12,5 г (выход 27%) того же самого продукта (показатель гомогенности при определении путем HPLC был равен 97,1%).Alternative procedure
Fragmented molecular sieves

placed in a 1-liter 1-necked flask dried above the flame, which was then connected to a vacuum line (approximately 0.5 mm Hg). The sieves were heated using a hot air dryer for about 10 minutes with manual shaking. After cooling under vacuum, argon was introduced into the flask and 2'-O- (ethoxycarbonyl) -7-O-methylthiomethylpaclitaxel (37.5 g, 38.03 mmol) was added, followed by dibenzyl phosphate (14.3 g, 53.24 mmol) and tetrahydrofuran (400 ml). The heterogeneous mixture was vigorously stirred for 15 minutes at room temperature using a magnetic stirrer. In a separate flame-dried flask, N-iodosuccinimide (10.7 g, 47.54 mmol) was dissolved in tetrahydrofuran (50 ml) under argon. (During the preparation of the NIS fluid transfer solution and the course of the reaction, the vessels were coated with aluminum foil to protect from light). Then, the prepared solution was slowly added via syringe to the reaction mixture (10 min). The flask containing NIS was washed with 5 ml of THF and transferred to the reaction mixture, which was then stirred for 2 hours at room temperature. Analysis by thin layer chromatography showed the absence of starting material. The dark red solution was filtered through a celite pad directly into a vigorously stirred biphasic mixture containing ethyl acetate (500 ml) 10% aqueous sodium thiosulfate solution (300 ml) and saturated sodium bicarbonate solution (200 ml). After a few seconds, the red color disappeared and the solution became colorless. The celite layer was washed with ethyl acetate (approximately 100 ml) and both layers of the liquid were transferred to a separatory 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 NaHCO ₃ and 5% K ₂ CO ₃ and then saturated NaHCO ₃ (2 x 500 ml), half-saturated brine (1 x 500 ml) and brine (1 x 500 ml). The extract was dried with anhydrous MgSO ₄ and filtered. Then it was treated with 5.0 g of neutral norite (charcoal) by stirring at room temperature for 15 minutes. Again it was filtered through a pad of celite, and by removing the solvent under reduced pressure, 52 g of crude product was obtained. It was dissolved in a mixture of toluene and methylene chloride (280 ml / 25 ml), and hexanes (20 ml) were added dropwise to the resulting solution. After 3 hours at room temperature, the crystallized mixture was kept at 0 ^° C. overnight. A pale yellow solid formed on the walls of the flask. After decanting the mother liquor, the residue was triturated with toluene (50 ml), filtered, washed with toluene and dried on a vacuum filter for 30 minutes. Then it was transferred to a desiccator with dreiter, and then dried in vacuum (approximately 0.5 mm Hg) for 4 hours, resulting in 24.4 g (53% yield) of the title compound (homogeneity index when determining by HPLC was 95.9%). The mother liquor was evaporated to dryness, triturated with toluene (100 ml), filtered, washed with toluene and dried on a vacuum filter for 30 minutes. After drying in a desiccator, as described above, 12.5 g (27% yield) of the same product were obtained (homogeneity index determined by HPLC was 97.1%).

К раствору 2'-O-(этоксикарбонил)-7-O-дибензилфосфоно-оксиметилпаклитаксела (1,23 г, 1,01 ммоль) в сухом этилацетате (40 мл) добавляли 10%-ный палладий на угле (428 мг, 10%, 0,404 ммоль). Реакционную смесь подвергали гидрогенизации при 60 фунтах на кв. дюйм (400 кПа) с непрерывным встряхиванием в течение 24 часов. Твердое вещество отфильтровывали через целит, после чего целит несколько раз промывали этилацетатом. Фильтрат концентрировали и получили свободную кислотную форму указанного в заголовке с соединения (1,01 г, 80%-ная чистота при определении путем HPLC). На следующей стадии удаляли примеси путем препаративной хроматографии на колонке C-18.(d) Preparation of 2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel and its monosodium, monopotassium, triethylamine, arginine, lysine, ethanolamine, N-methylglucamine and triethanolamine salts

To a solution of 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphono-hydroxymethylpaclitaxel (1.23 g, 1.01 mmol) in dry ethyl acetate (40 ml) was added 10% palladium on carbon (428 mg, 10% 0.404 mmol). The reaction mixture was hydrogenated at 60 psi. inch (400 kPa) with continuous shaking for 24 hours. The solid was filtered through celite, after which the celite was washed several times with ethyl acetate. The filtrate was concentrated to give the free acid form of the title compound (1.01 g, 80% purity as determined by HPLC). In the next step, impurities were removed by preparative chromatography on a C-18 column.

Mass spectrum (FAB / NOBA, NaI, KI): [M + Na] ⁺ , m / z 1058; [M + K] ⁺ , m / z 1074; [M + 2Na - H] ⁺ , m / z 1080; [M + Na + K - H] ⁺ , m / z 1096; [M + 2K - H] ⁺ , m / z 1112.

High-resolution mass spectrum (FAB / NOBA, external CSI / Gli standard): [M + Na] ⁺ , m / z 1058, 3163
(C ₅₁ H ₅₈ NO ₂₀ PNa, calculated value: 1058, 3188; deviation Δ = 2.3 ppm).

¹ H NMR (acetone-d ₆ / D ₂ O), δ: 1.13 (3H, s), 1.21 (3H, s), 1.66 (3H, s), 1.87 (H, m ), 1.93 (3H, s), 2.14 (3H, s), 2.18 (H, m), 2.44 (3H, s), 2.95 (H, m), 3.81 (H, d), 4.12 (2H, s), 4.15 - 4.27 (3H, overlapping multiplets), 4.92 - 4.99 (2H, overlapping wide multiplets), 5.15 (H, bc), 5.48 (H, d), 5.61 (H, d), 5.84 (H, dd), 6.07 (H, t), 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).

Alternative procedure
2'-O- (ethoxycarbonyl) -7-O- (dibenzylphosphonooxymethyl) - paclitaxel (490 mg, 0.402 mmol) in ethyl acetate (20 ml) was hydrogenated in a Parr shaker at 60 psi. inch (400 kPa) in the presence of palladium on charcoal (10%, mass / mass, 150 mg). Control was carried out by thin layer chromatography and high pressure liquid chromatography (HPLC). When no further starting material or intermediate (preferably monobenzyl phosphate) was detected (26 h), the suspension was filtered through celite and evaporated to dryness. HPLC analysis showed a purity of 88 - 92%.

Alternative procedure
The 2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel triethylamine salt described below (5.4 g, 4.75 mmol) was intensively partitioned between ethyl acetate (100 ml) and 5% NaHSO ₄ (45 ml) with stirring at 0 ^o C for 30 minutes The aqueous layer was separated and extracted with ethyl acetate (20 ml). The combined ethyl acetate layer was washed with semi-saturated brine (25 ml), brine (25 ml x 2), dried over NaSO ₄ and filtered, resulting in a solution of an acid (about 4.75 mmol) in ethyl acetate (about 150 ml). This solution was then concentrated to dryness on a rotary evaporator to give 3.75 g of the title compound as a free acid form in 95% yield.

 Monosodium salt was prepared as follows.

A sample of 2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel (1.6 g, 1.55 mmol) was dissolved in acetonitrile (30 ml) by ultrasonic treatment. This solution was diluted with water (30 ml) and 1.1 M NaHCO ₃ solution (2.11 ml, 2.32 mmol) was added to it, shaken alternately and sonicated to obtain a solution (5 - 20 min). A slightly milky solution was applied to a C-18 column, washed with water in an amount of two column volumes, and then the monosodium salt of 25% acetonitrile in water was eluted. The appropriate fractions were combined, acetonitrile was evaporated and the aqueous phase was lyophilized to give the monosodium salt of the title compound (850 mg, about 50%) with a purity of 97% as determined by HPLC.

Mass spectrum (FAB / NOBA, NaI, KI); [M + Na] ⁺ , m / z 1180.

High-resolution mass spectrum (FAB / NOBA, external CSI / Gli standard): [M + Na] ⁺ , m / z 1080, 2968 (C ₅₁ H ₅₇ NO ₂₀ PNa ₂ ), calculated value: 1080, 3007; deviation D = 3.6 ppm).

 Elemental analysis: C 52.65 (calc. 56.72), H 5.06 (calc. 5.23), N 1.20 (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 .

¹ H NMR (DMSO-d ₆ , D ₂ O-acetone-d ₆ ), δ: 1.10 (6H, s), 1.23 (3H, t), 1.64 (3H, s), 1, 70 (H, m) 1.90 (3H, s), 1.99 (H, m), 2.14 (3H, s), 2.37 (3H, s), 2.98 (H, m) , 3.74 (H, d), 4.07 (2H, s), 4.13 - 4.26 (3H, m, overlapping), 4.80 (H, bdd), 4.97 (H, d ), 5.09 (H, pcs), 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), 8.06 (2H, dd).

 Sodium salt was prepared as follows.

The crude 2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel (89%; 70 mg, 0.060 mmol) in ethyl acetate (2 ml) was treated with a solution of sodium ethyl hexanoate (87.5 mmol in ethyl acetate, 1.0 ml, 0, 0875 mmol) at room temperature with stirring. After stirring at room temperature for 1 h, hexane (1.2 ml) was added to the cloud point. After standing at -20 ^° C for 2 hours, a fine amorphous powder was filtered (with some difficulty, very slowly) through a thin filter paper and 45 mg (70%) of sodium salt were obtained. It had a purity (determined by HPLC) of 95.2% and contained a small amount of ethyl hexanoic acid (NMR).

 The triethanolamine salt was prepared as follows.

The resulting hydrogenation (89% by HPLC 2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylapaclitaxel (0.69 g, 0.593 mmol after adjusting for impurities) was dissolved in ethyl acetate (10 ml) and stirred slowly, while adding a solution of triethanolamine (0.11 M in ethyl acetate, using 5.1 ml, 0.95 equiv.) was added dropwise. The milky-colored solution obtained in this procedure was digested at 0 ^° C for 2 hours, after which it was filtered through a thin filter paper, washing with cold ethyl acetate, 499 mg (80%) of an amorphous fine non-electrostatic was obtained powder, which was dried overnight in vacuo. HPLC showed 96.6% purity (C-18, 45% 5 mM Q ₁₂ + 10 mM ammonium phosphate with a pH of 6.55% acetonitrile). NMR spectrum (D ₂ O / acetone / DMSO) showed traces of ethyl acetate and the absence of other clearly defined impurities. An analysis was made on 2-3 hydrates.

 Then, the triethanolamine salt obtained previously from another experiment was purified as follows. The triethanolamine salt (approximately 2 g) was dissolved in approximately 30% acetonitrile in water. This solution was eluted with a slight nitrogen pressure through a C-18 column (Bakerbond) with a gradient of 20-40% acetonitrile in water. Fractions containing the desired triethanolamine salt were captured and acetonitrile was removed by rotary evaporation under reduced pressure. Aqueous solutions were frozen and lyophilized overnight, resulting in 1.4 g of triethanolamine salt with a purity of 97.5%.

 Triethanolamine salt can also be obtained as follows.

2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel triethylamine salt (3.0 g, 2.64 mmol) was partitioned between ethyl acetate (60 ml) and 5% NaHSO ₄ (30 ml) with vigorous stirring at 0 ^o C for 15 minutes. The aqueous layer was separated and extracted with ethyl acetate (10 ml). The combined ethyl acetate layer was washed with brine (15 ml), dried over Na ₂ SO ₄ , filtered, and a solution of an acid (about 2.64 mmol) in ethyl acetate (about 70 ml) was obtained. To this solution, N (CH ₂ CH ₂ OH) ₃ (0.35 ml, 2.64 mmol) was added dropwise under vigorous stirring at room temperature over 5 minutes. The resulting suspension was stirred for another 1 h, after which it was filtered, washed with ethyl acetate (15 ml × 2), dried in vacuo, and 2.8 g of triethanolamine salt was obtained in 89% yield. HPLC analysis showed a homogeneity index of 98.7%; mp: above 157 ^o C, with decomposition.

Elemental analysis: calculated for C ₅₆ H ₇₃ N ₂ O ₂₃ P • 2.0 H ₂ O • 0.3 ethyl acetate: C 55.60; H 6.48; N, 2.27; KF (H ₂ O) 2.92. Found: C, 55.94; H 6.59; N, 2.43; KF (H ₂ O) 3.50.

 Triethylamine salt was prepared as follows.

To a solution of 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (10 g, 8.23 mmol) in ethyl acetate (350 ml) was added 10% palladium-carbon (2 g, 20% charge) at room temperature . The resulting suspension was degassed by suction of air, followed by purging with argon. This process was repeated two more times. Then the argon was replaced with hydrogen, after which the same degassing procedure was carried out. The resulting suspension was stirred under hydrogen pressure from a balloon (2-3 psi, i.e. 0.14-0.21 kg / cm ² ) for 16 hours at room temperature with vigorous stirring. Hydrogen was aspirated and replaced with argon three times, followed by a degassing procedure. The resulting suspension was filtered through a pad of celite. Et ₃ N (8.23 mmol, 1.14 ml) was slowly added to this homogeneous filtrate over 5 minutes with vigorous stirring. The resulting finely divided white suspension was stirred for another 30 minutes. It was filtered through a funnel with a sintered porous filter with porosity E. The filtration residue was dried in vacuo (1 mmHg) for 16 hours to give 8.22 g of the title triethylamine salt in 88% yield. HPLC analysis gave a homogeneity index of 97.4%; mp: above 178 ^o C with decomposition.

Elemental analysis: calculated for C ₅₇ H ₇₃ N ₂ O ₂₀ P • 4.5 H ₂ O: C 56.19; H 6.79; N, 2.30; KF (H ₂ O) 6.65.

Calculated: C 56.33; H 6.87; N, 2.32; KF (H ₂ O) 7.96.

 An alternative procedure for obtaining triethylamine salt.

 To a 250 ml flask was added 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (5.67 g, 4.66 mmol) and dissolved in ethyl acetate (150 ml). The flask was equipped with a three-way valve, one of the connections of which was connected to the vacuum line, and the other to the argon line. By using a valve, a partial vacuum was created in the flask and then purged with argon. This process was repeated two more times. Activated carbon palladium (10% Pd) (0.85 g) was added to the flask. The argon line attached to the three-way valve was replaced with a cylinder filled with hydrogen. By means of a valve, a vacuum was created in the flask, and then it was purged with hydrogen. This process was repeated four more times. The resulting mixture was stirred at room temperature in a hydrogen atmosphere from a balloon overnight. Analysis by thin layer chromatography 17 hours after the initial exposure to hydrogen showed the absence of starting material. A hydrogen cylinder attached to a three-way valve was replaced with an argon line. By means of a valve, a partial rarefaction was created in the flask, after which it 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 (2 x 10 ml). Triethylamine (0.650 ml, 4.66 mmol) was added to the stirred filtrate. The resulting suspension was stirred at room temperature for two hours, after which the volume was reduced to about 150 ml by evaporation on a rotary evaporator. The solid was filtered off, washed with ethyl acetate (2 x 10 ml) and dried in vacuo to yield 4.76 g (90% yield) of the title triethylamine salt as a white powder (HPLC analysis showed a homogeneity index of 96, 6%).

 An alternative procedure for obtaining triethylamine salt.

 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (5.17 g, 4.25 mmol) was introduced into a 250 ml flask and dissolved in ethyl acetate (150 ml). The flask was equipped with a three-way valve, one of the connections of which was connected to the vacuum line, and the other to the argon line. Using the valve, a partial vacuum was created in the flask, after which it was purged with argon. This process was repeated two more times. Palladium on activated carbon (10% Pd, 0.86 g) was added to the flask. The argon line attached to the three-way valve was replaced with a cylinder filled with hydrogen. By means of a valve, a partial rarefaction was created in the flask, after which it was purged with hydrogen. This process was repeated five more times. The resulting mixture was stirred at room temperature in a hydrogen atmosphere from a balloon overnight. Analysis by thin layer chromatography 16 hours after the initial exposure to hydrogen showed the absence of starting material. A hydrogen cylinder attached to a three-way valve was replaced with an argon line. By means of a valve, a partial vacuum was created in the flask, after which it 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). Triethylamine (0.590 ml, 4.25 mmol) was added to the stirred filtrate. The resulting suspension was stirred at room temperature for one hour, after which the volume was reduced to about 140 ml by evaporation on a rotary evaporator. The solid was filtered off, washed with ethyl acetate (10 ml) and dried under vacuum, yielding 4.46 g (92% yield) of the title triethylamine salt as a white powder (homogeneity determined by HPLC analysis was 96.7%).

 The lysine salt was prepared as follows.

To a suspension of 10% palladium on carbon (20% loading, 3 g) in ethanol (600 ml, strength 200) at 0 ^° C., 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethyl paclitaxel (15.0 g, 12.34 mmol). The resulting suspension was degassed by suctioning air and purging with argon. This process was repeated two more times. Then the argon was replaced with hydrogen, after which the same degassing procedure was carried out with vigorous stirring at 0 ^° C. for 2 hours. The cooling bath was removed and the reaction mixture was stirred at ambient temperature for another 4.5 hours. The reaction mixture was degassed by suction of hydrogen and purging with argon three times, 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, strength 200, 20 ml) over 5 minutes with vigorous stirring. Distilled water (110 ml) was added to the resulting white suspension, and the suspension was stirred for 30 minutes. It was heated to about 55 ^o C. The resulting homogeneous solution was kept in an oil bath at 50 ^o C and slowly cooled to room temperature for 16 hours and to 4 ^o C for 3 hours, filtered and subjected to vacuum drying for 16 hours obtained 11.8 g (yield about 80%) of a lysine salt having a uniformity index of 99.0% as determined by HPLC; so pl. above 170 ^o C with decomposition.

Elemental analysis: calculated for C ₅₇ H ₇₂ N ₃ O ₂₂ P • 8.0 H ₂ O:
C 51.62; H 6.69; N, 3.17; KF (H ₂ O) 10.87.

Found: C, 51.76; H 6.57; N 3.48; KF (H ₂ O) 11.42.

Ethanolamine salt was prepared as follows.
2'-O- (ethoxycarbonyl) -7-O-phosphonooxymethylpaclitaxel triethylamine salt (3.0 g, 2.64 mmol) was partitioned between ethyl acetate (60 ml) and 5% NaHSO ₄ (30 ml) with vigorous stirring at 0 ^o C for 15 minutes. The aqueous layer was separated and extracted with ethyl acetate (15 ml). The combined ethyl acetate layer was washed with brine (15 ml), dried over Na ₂ SO ₄ , filtered, and a solution of the free acid (about 2.64 mmol) in ethyl acetate (about 70 ml) was obtained. To this solution, a solution of H ₂ NCH ₂ CH ₂ OH (0.15 ml, 2.64 mmol) in ethyl acetate (5 ml) was added dropwise with vigorous stirring at room temperature over 5 minutes. The resulting suspension was stirred for another 1 hour, after which it was filtered, washed with ethyl acetate (15 ml x 2) and dried in vacuo, whereby 2.6 g of the title ethanolamine salt was obtained in 89% yield. The uniformity index when determined by HPLC analysis was equal to 97.8%; so pl. above 130 ^o C with decomposition.

Elemental analysis: calculated for C ₅₃ H ₆₅ N ₂ O ₂₁ P • 2.5 H ₂ O:
C 55.73; H 6.18; N, 2.45; KF (H ₂ O) 3.94.

Found: C, 55.76; H 6.39; N, 2.45; KF (H ₂ O) 6.00.

 Arginine salt was prepared as follows.

To a suspension of 10% palladium on carbon (loading 20%, 6 g) in ethanol (900 ml, strength 200) at 0 ^° C., 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (30.0 g 24.69 mmol). The resulting suspension was degassed by suctioning air and purging with argon. This process was repeated two more times. Then, argon was replaced with hydrogen, after which again the degassing procedure described above was carried out with vigorous stirring. The resulting mixture was stirred at 0 ^° C. for 2 hours. The cooling bath was removed and the reaction solution was stirred at ambient temperature for another 24 hours. The reaction mixture was degassed by aspirating hydrogen and purging with argon, performing the above procedure three times. It was filtered under argon through a layer of celite. The filtrate was divided into two equal parts, and ethanol (190 ml, strength 200) was added to each part. To one part (approximately 630 ml), a solution of arginine (2.0 g, 0.94 equiv.) In a mixture of water: ethanol (2: 1, strength 200, 20 ml) was slowly added over 5 minutes with vigorous stirring. Distilled water (100 ml) was added to the resulting white suspension and stirred for 30 minutes, after which it was heated to about 60 ^° C. It was filtered hot and the filtrate was kept in an oil bath at 50 ^° C., allowing it to cool to room temperature, and kept at room temperature for 2 hours and at 4 ^o C for 2 hours. It was filtered and washed with a cold mixture of 3% water in ethanol (100 ml) and dried by suction for 16 hours, resulting in 12.95 g (approximately 86% yield) of the title arginine salt with a uniformity index of 96 , 7%.

This material (12.95 g) was dissolved in a mixture of 15% water in ethanol (approximately 700 ml) at 55 ^o C. The solution was cooled and kept at 30 ^o C for 3.5 hours, at room temperature for 16 hours and at 4 ^o C for 3 hours. The resulting crystals were filtered off, washed with a cold mixture of 2% water in ethanol (50 ml × 2), dried by suction for 4 hours, and then dried in vacuum (1 mmHg) for 16 hours, resulting in 10.2 g (80% yield) of the title arginine salt (uniformity index 98.5%); mp: above 176 ^o C with decomposition.

Elemental analysis: calculated for C ₅₇ H ₇₂ N ₅ O ₂₂ P • 6.4 H ₂ O:
C 51.65; H 6.45; N, 5.28; KF (H ₂ O) 8.7.

Found: C, 51.86; H 6.65; N 5.53; KF (H ₂ O) 8.72.

 N-methylglucamine salt was prepared as follows.

To a suspension of 10% palladium on charcoal (20% charge, 6 g) in methanol (900 ml, strength 200) at 0 ^° C., 2'-O- (ethoxycarbonyl) -7-O-dibenzylphosphonooxymethylpaclitaxel (30.0 g, 24.69 mmol). The resulting suspension was degassed by suctioning air and purging with argon. This process was repeated two more times. Then, argon was replaced with hydrogen, after which the above procedure was carried out with vigorous stirring. The resulting mixture was stirred at 0 ^° C. for 2 hours. The cooling bath was removed and the reaction solution was stirred at ambient temperature for another 24 hours. The reaction mixture was degassed by aspirating hydrogen and purging with argon, following the above procedure three times. It was filtered under argon through a layer of celite. The filtrate was divided into two equal parts, and ethanol (190 ml, strength 200) was added to each part. To one part (approximately 600 ml), a solution of N-methylglucamine (2.24 g, 0.94 equiv.) In a mixture of water: ethanol (1: 1, strength 200, 20 ml) was slowly added over 5 minutes with vigorous stirring. Distilled water (100 ml) was added to the resulting white suspension, and the suspension was stirred for 30 minutes and then heated to about 49 ^° C. A clear homogeneous solution was kept in an oil bath at 50 ^° C, allowing it to cool to room temperature, and kept at room temperature for 2 hours and at 4 ^o C for 1.5 hours. The solution was filtered, washed with a mixture of 3% water in ethanol, and dried by suction at room temperature for 16 hours, resulting in 9.65 g (approximately 64% yield) of the title N-methylglucamine salt with a uniformity index of 96.4%.

This material (9.65 g) was dissolved in a mixture of 15% water in ethanol (approximately 450 ml) at 52 ^o C. Then the solution was cooled and kept at 28 ^o C for 3.5 hours, at room temperature for 16 hours and at 4 ^o C 3 hours. The obtained crystals were filtered off, washed with a cold mixture of 2% water in ethanol (50 ml × 2), dried by suction for 4 hours, and then dried under vacuum (1 mmHg), resulting in 7.5 g (approximately 80% yield) of the title N-methylglucamine salt (the uniformity index when determined by HPLC was 98.6%); mp: above 154 ^o C with decomposition.

Elemental analysis: calculated for C ₅₈ H ₇₅ N ₂ O ₂₅ P • 5.0 H ₂ O:
C 52.72; H 6.48; N, 2.12; KF (H ₂ O) 6.82.

Found: C, 53.09; H 6.50; N, 2.08; KF (H ₂ O) 7.12.

К охлажденному (0 - (-5^oC)) раствору 7-O-(триэтилсилил)паклитаксела (2,46 г, 2,5439 ммоль) в сухом ацетонитриле (100 мл) добавляли диметилсульфид (1,348 г, 1,59 мл, 21,6976 ммоль), а затем бензоилпероксид (2,628 г, 10,8488 ммоль). Гетерогенную смесь перемешивали при 0^oC 1 час и держали при 5^oC 18 часов. Получили желтый раствор. Его упаривали досуха и очищали на колонке с силикагелем (элюирование смесью этилацетат:гексан с отношением 1:4, 1: 3 и 1:2), получив в результате указанное в заголовке соединение (1,0 г, 38%). Оно было использовано как таковое для следующей стадии.Example 4. 2'-O- (phosphonooxymethyl) paclitaxel
(a) Preparation of 2'-O- (methylthiomethyl) -7-O- (triethylsilyl) paclitaxel

To a cooled (0 - (-5 ^° C)) solution of 7-O- (triethylsilyl) paclitaxel (2.46 g, 2.5439 mmol) in dry acetonitrile (100 ml) was added dimethyl sulfide (1.348 g, 1.59 ml, 21.6976 mmol) and then benzoyl peroxide (2.628 g, 10.8488 mmol). The heterogeneous mixture was stirred at 0 ^° C. for 1 hour and kept at 5 ^° C. for 18 hours. Got a yellow solution. It was evaporated to dryness and purified on a silica gel column (eluted with ethyl acetate: hexane 1: 4, 1: 3 and 1: 2) to give the title compound (1.0 g, 38%). It was used as such for the next step.

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

К охлажденному (-15^oC) раствору полученного на стадии (a) продукта (1,0 г, 0,9737 ммоль) в сухом ацетонитриле (30 мл) добавляли пол каплям 0,5 н. HCl (3 мл). Полученный раствор перемешивали при -15^oC в течение 1 часа и при 5^oC в течение 18 часов. Разбавляли его этилацетатом (20 мл) и промывали холодным 6%-ным раствором NaHCPО₃ и рассолом. Высушивали (МgSO₄) и упаривали досуха. Полученный продукт очищали на покрытой силикагелем пластинке (метиленхлорид 15% ацетонитрила) и получали чистое указанное в заголовке соединение (280 мг, 31,4%).(b) Preparation of 2'-O- (methylthiomethyl) paclitaxel

To a cooled (-15 ^° C) solution of the product obtained in step (a) (1.0 g, 0.9737 mmol) in dry acetonitrile (30 ml) was added half a drop of 0.5 N HCl (3 ml). The resulting solution was stirred at -15 ^° C for 1 hour and at 5 ^° C for 18 hours. It was diluted with ethyl acetate (20 ml) and washed with cold 6% NaHCPO ₃ solution and brine. Dried (MgSO ₄ ) and evaporated to dryness. The resulting product was purified on a silica gel-coated plate (methylene chloride 15% acetonitrile) to give the pure title compound (280 mg, 31.4%).

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

NMR (CDCl ₃ ): δ 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.

К перемешиваемому раствору полученного на стадии (b) продукта (0,89 г, 0,9748 ммоль) в сухом 1,2-дихлорэтане (12 мл) добавляли порошкообразные

молекулярные сита (1,0 г), а затем добавляли по каплям раствор смеси N-иодсукцинимида (0,33 г, 1,4622 ммоль) и дибензилфосфата (0,41 г, 1,4622 ммоль) в сухом тетрагидрофуране (8 мл). Полученную смесь перемешивали при комнатной температуре в течение 1 часа, после чего фильтровали через целит. Фильтрат упаривали досуха, и красный остаток растворяли в этилацетате (50 мл) и промывали холодным 6%-ным NaHSO₄, холодным 6%-ным NaHCO₃ рассолом. Раствор высушивали (MgSO₄) и упаривали, получив в результате пену, ее очищали посредством покрытой силикагелем пластины (метиленхлорид-20% ацетонитрила) и получали чистый продукт (0,77 г, 69%).High-resolution mass spectrum: MH ⁺ - 914.3394 (calculated 914.3422)
(c) Preparation of 2'-O- (dibenzylphosphonooxymethyl) paclitaxel

To a stirred solution of the product obtained in step (b) (0.89 g, 0.9748 mmol) in dry 1,2-dichloroethane (12 ml) was added powdered

molecular sieves (1.0 g), and then a solution of a mixture of N-iodosuccinimide (0.33 g, 1.4622 mmol) and dibenzyl phosphate (0.41 g, 1.4622 mmol) in dry tetrahydrofuran (8 ml) was added dropwise. . The resulting mixture was stirred at room temperature for 1 hour, after which it was 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% NaHSO ₄ , cold 6% NaHCO ₃ brine. The solution was dried (MgSO ₄ ) and evaporated, resulting in a foam, it was cleaned with a silica gel plate (methylene chloride-20% acetonitrile) and a pure product was obtained (0.77 g, 69%).

IR spectrum: 3854, 3744, 3362, 3066, 1960, 1722, 1602, 1580 cm ^-1.
NMR (CDCl ₃ ): δ 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.

Смесь полученного на стадии (с) продукта (0,9 г, 0,7874 ммоль) и 10% Рd/C (1 г) в этилацетате (20 мл) гидрогенизировали при 60 фунтах на кв. дюйм (400 кПа) в течение 24 часов. Реакционную смесь фильтровали через целит, и фильтрат упаривали досуха. Остаток очищали посредством покрытой силикагелем пластины (метиленхлорид-40% метанола) и получили указанный в заголовке продукт (0,254 г, 33,4%);
т.пл. 202 - 205^oC (разложение).(d) Preparation of 2'-O- (phosphonooxymethyl) paclitaxel

A mixture of the product obtained in step (c) (0.9 g, 0.7874 mmol) and 10% Pd / C (1 g) in ethyl acetate (20 ml) was hydrogenated at 60 psi. inch (400 kPa) for 24 hours. The reaction mixture was filtered through celite, and the filtrate was evaporated to dryness. The residue was purified through a silica gel-coated plate (methylene chloride-40% methanol) to give the title product (0.254 g, 33.4%);
so pl. 202 - 205 ^o C (decomposition).

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

NMR (acetone-d ₆ / D ₂ O): δ 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 + KH] ⁺ , 1024; [M + 2K-H] ⁺ , 1040.

High-resolution mass spectrum: MNa ⁺ - 986.2955 (calculated 986.2976).

К перемешанному раствору паклитаксела (0,853 г, 1 ммоль) и диметилсульфида (1,465 г, 20 моль) в ацетонитриле (20 мл) при 0^oC добавляли твердый бензоилпероксид (1,995 г, 8 ммоль). Реакционную смесь интенсивно перемешивали при 0^oC в течение 3 часов. Ход реакции контролировали путем тонкослойной хроматографии в смеси гексан:этилацетат (1:1, объем/объем), R_f паклитаксела = 0,24, R_f продукта = 0,60. После исчезновения исходного материала (приблизительно через 3 часа) реакцию быстро прекращали путем выпаривания растворителей досуха при 25^oC с использованием вакуумной линии в помещении. Сухой остаток разделяли посредством колонки с силикагелем (EM Science, 40 - 63 мкм), 100 мл сухого силикагеля, размер колонки : Ф = 3/4 дюйма (19 мм), система растворителей: гексан: этилацетат (3:2, объем/объем), объем каждой фракции: приблизительно 25 мл. Из фракции 15 - 19 было извлечено указанное в заголовке соединение (0,515 г, 53%-ный выход).Example 5. 2 ', 7-O- (Phosphonoxymethyl) paclitaxel sodium salt
(a) Preparation of 2 ', 7-O-bis (methylthiomethyl) paclitaxel

Solid solution of benzoyl peroxide (1.995 g, 8 mmol) was added to a mixed solution of paclitaxel (0.853 g, 1 mmol) and dimethyl sulfide (1.465 g, 20 mol) in acetonitrile (20 ml) at 0 ^° C. The reaction mixture was vigorously stirred at 0 ^° C. for 3 hours. The progress of the reaction was monitored by thin layer chromatography in hexane: ethyl acetate (1: 1, v / v), R _f paclitaxel = 0.24, R _f product = 0.60. After the disappearance of the starting material (after about 3 hours), the reaction was quickly stopped by evaporating the solvents to dryness at 25 ^° C. using an indoor vacuum line. The dry residue was separated by silica gel column (EM Science, 40 - 63 μm), 100 ml of dry silica gel, column size: Ф = 3/4 inch (19 mm), solvent system: hexane: ethyl acetate (3: 2, volume / volume) ), the volume of each fraction: approximately 25 ml. The title compound (0.515 g, 53% yield) was recovered from fraction 15-19.

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): λ _max = 204 nm, E (1% / 1 cm) = 243.45; λ _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 .

¹ H NMR (CDCl ₃ ), δ: 1.17 (3H, s), 1.20 (3H, s), 1.68 (3H, s), 1.74 (3H, s), 1.84 ( H, dd), 2.04 (3H, d), 2.09 (3H, s), 2.15 (3H, s) overlaps with (H, m), 2.37 (H, dd), 2, 51 (3H, s), 2.79 (H, ddd), 3.78 (H, d), 4.18 (H, d), 4.28 (H, m), 4.31 (H, d ), 4.53 - 4.74 (4H, two overlapping AB m), 4.93 (H, d), 4.95 (H, d), 5.68 (H, d), 5.82 (H dd), 6.24 (H, dd), 6.54 (H, s), 7.05 (H, d), 7.28 - 7.59 (10H, overlapping m), 7.57 (H , m), 7.76 (2H, d), 8.09 (2H, d).

К смеси 2', 7-O-бис(метилтиометил)пакклитаксела (198 мг, 0,2 ммоль) и

молекулярных сит (примерно 200 мг) в метиленхлориде (12 мл) при комнатной температуре добавляли раствор N-иодсукцинимида (135 мг, 0,5 ммоль) и дибензилфосфата (167 мг, 0,5 ммоль) в сухом тетрагидрофуране (8 мл). Реакционную смесь перемешивали 1,5 часа затем молекулярные сита офильтровывали на целите, промывали метиленхлоридом (10 мл), и выпаривали растворители досуха при комнатной температуре с использованием бытовой вакуумной линии. Остаток растворяли в этилацетате (100 мл) и промывали в делительной воронке 1%-ным тиосульфатом натрия (50 мл), 0,5 м бикарбонатом натрия (50 мл) и дважды водой (2 х 50 мл). Органическую фазу высушивали над сульфатом магния, упаривали досуха и вновь растворяли в этилацетате (1 мл). Продукт осаждали 50 мл смеси диэтиловый эфир: гексан (1:1) и дважды промывали одной и той же системой растворителей (2 х 50 мл). Получили сырой продукт (218 мг) с 74%-ным выходом. Очистку этого продукта осуществляли путем загрузки его раствора в метиленхлориде (3 мл) на силикагель (Ф = 3/4 дюйма (19 мм) х L = 1 дюйм (25,4 мм)) и элюирования продукта 50 мл смеси метиленхлорид: этилацетат (3: 1). Получили указанное в заголовке соединение (172,7 мг) с выходом 59,3%.(b) Preparation of 2 ', 7-O- (dibenzylphosphonooxymethyl) paclitaxel

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, a solution of N-iodosuccinimide (135 mg, 0.5 mmol) and dibenzyl phosphate (167 mg, 0.5 mmol) in dry tetrahydrofuran (8 ml) was added. The reaction mixture was stirred for 1.5 hours, then the molecular sieves were filtered off on Celite, washed with methylene chloride (10 ml), and the solvents were evaporated to dryness at room temperature using a household vacuum line. The residue was dissolved in ethyl acetate (100 ml) and washed in a separatory 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 redissolved in ethyl acetate (1 ml). The product was precipitated with 50 ml of a mixture of diethyl ether: hexane (1: 1) and washed twice with the same solvent system (2 x 50 ml). A crude product (218 mg) was obtained in 74% yield. This product was purified by loading its solution in methylene chloride (3 ml) onto silica gel (Ф = 3/4 inch (19 mm) x L = 1 inch (25.4 mm)) and eluting the product with 50 ml of a mixture of methylene chloride: ethyl acetate (3 : 1). The title compound (172.7 mg) was obtained in 59.3% yield.

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

UV spectrum (MeCN): λ _max = 194 nm, E (1% / 1 cm) = 1078.36; λ _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 .

¹ H-NMR (CDCl ₃ ), δ: 1.12 (3H, s), 1.14 (3H, s), 1.56 (H, m), 1.67 (3H, s), 1.84 (3H, d), 1.90 (H, m), 2.17 (3H, s), 2.29 (3H, s), 2.73 (H, m), 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 (H, dd), 5.38 (H, dd), 5.54 - 5.64 (2H, overlapping m), 5.99 (H, br. dd), 6.25 (H, s), 7.11 - 7.14 (2H, m), 7.24 - 7.64 (28H, overlapping m), 7.94 (2H, dd), 8, 04 (2H, dd), 8.30 (H, d).

Пробу 2', 7-O-бис(дибензилфосфонооксиметил)паклитаксела (112 мг, 0,078 ммоль) растворяли в этилацетате (7 мл) и гидрогенизировали на 10% палладия на угле (50 мг) при комнатной температуре и 60 фунтах на кв. дюйм (400 кПа) в течение 2 часов. Катализатор удаляли путем фильтрования на целите. Целит промывали этилацетатом (10 мл). Фильтрат обрабатывали твердым бикарбонатом натрия (20 мг, 3 эквив.), после чего выпаривали растворитель досуха. Сухой остаток вновь растворяли в 5 мл смеси вода: ацетон (4:1, объем/объем) и очищали путем хроматографии на колонке C-18 с обратной фазой (55 - 105 мк C-18, Уотерс, 50 мл сухого C-18, Ф = 3/4 дюйма (19 мм), в смеси вода: ацетон (4: 1, объем/объем). Элюент на аналитической колонке C-18 Джонса для НPL C (15 см, 1 мл/мин, λ = 230 нм) в буфере ацетонитрил: фосфат с pH 6 (50/50, объем/объем) с добавлением коктейля Q12 из ионных пар (Regis), R_t = 4,7 минут. Фракции, содержащие указанный в заголовке продукт, объединяли, выпаривали ацетон под бытовым вакуумом при 20^oC, и раствор лиофилизировали. Был получен указанный в заголовке продукт (44,2 мг) с выходом 58,8%.(c) Preparation of 2 ', 7-O-bis (phosphonooxymethyl) paclitaxel sodium salt

A sample of 2 ', 7-O-bis (dibenzylphosphonooxymethyl) paclitaxel (112 mg, 0.078 mmol) was dissolved in ethyl acetate (7 ml) and hydrogenated with 10% palladium-carbon (50 mg) at room temperature and 60 psi. inch (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.), After which the solvent was evaporated to dryness. The dry residue was redissolved in 5 ml of a water: acetone mixture (4: 1, v / v) and purified by chromatography on a reverse phase C-18 column (55 - 105 μm C-18, Waters, 50 ml of dry C-18, Ф = 3/4 inch (19 mm), in a mixture of water: acetone (4: 1, volume / volume). Eluent on an analytical column C-18 Jones for НPL C (15 cm, 1 ml / min, λ = 230 nm ) in acetonitrile buffer: phosphate, pH 6 (50/50, v / v) with the addition of a Q12 cocktail of ion pairs (Regis), R _t = 4.7 minutes, The fractions containing the title product were combined, the acetone was evaporated under domestic vacuum at 20 ^o C, and the solution was lyophilized. the title product (44.2 mg) was obtained in 58.8% yield.

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

UV spectrum (MeCN): λ _max = 192 nm, E (1% / 1 cm) = 129.73; λ _max = 230 nm, E (1% / 1 cm) = 26.43.

IR Spectrum (KBr): 3430, 3066, 2956, 1724, 1658, 1604, 1582, 1520, 1486, 1452, 1374, 1316, 1256, 1152, 1110, 1070, 1026, 966, 914, 802, 772, 710 , 538 cm ^-1 .

¹ H-NMR (acetone-d ₆ / D ₂ O), δ: 0.97 (3H, s), 1.02 (3H, s), 1.47 (H, m), 1.54 (3H, s), 1.70 (H, m), 1.75 (3H, s), 1.85 (H, m), 2.11 (3H, s), 2.30 (3H, s), 2, 88 (H, m), 3.64 (H, d), 4.03 (H, m), 4.06 (H, d), 4.16 (H, d), 4.74 (H, m) ), 4.86 (H, m), 51.11 (H, pcs), 5.22 (H, d), 5.42 (H, d), 5.90 (H, pcs), 6.21 (H, s), 7.06 (H, pc), 7.32 - 7.69 (10H, overlapping m), 7.80 (2H, d), 7.93 (2H, d).

К раствору 2'-O-этилоксикарбонил-7-O-метилтиометилпактилтаксела (соединение примера 3 (b), 27 г, 27,4 ммоль) в 100 мл тетрагидрофурана и 500 мл метанола добавляли свежеизмельченный K₂CO₃ (2,7 г, 19 ммоль). Раствор перемешивали 30 минут и нейтрализовали смолой IP-120 (Н⁺), фильтровали и концентрировали. Затем неочищенный фильтрат растворяли в 200 мл дихлорметана и перемешивали 24 часа с тетрабутиламмонийборогидридом (10 г). Раствор разбавляли дихлорметаном и промывали водой, насыщенным раствором бикарбоната и рассолом. После этого органическую фракцию высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (1:1, гексан:этилацетат) и получили 9,4 г указанного в заголовке соединения (53%) с точкой плавления 269^oC.Example 6. 7-O-Methylthiolmethylbaccatin III

To a solution of 2'-O-ethyloxycarbonyl-7-O-methylthiomethylpactyltaxel (compound of Example 3 (b), 27 g, 27.4 mmol) in 100 ml of tetrahydrofuran and 500 ml of methanol was added freshly ground K ₂ CO ₃ (2.7 g, 19 mmol). The solution was stirred for 30 minutes and neutralized with IP-120 (H ⁺ ) resin, filtered and concentrated. The crude filtrate was then dissolved in 200 ml of dichloromethane and stirred 24 hours with tetrabutylammonium borohydride (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 MgSO ₄ and concentrated. The residue was chromatographed on silica gel (1: 1, hexane: ethyl acetate) to obtain 9.4 g of the title compound (53%) with a melting point of 269 ^° C.

Mass spectrum FAB (NOBA) M + H calculated for C ₃₃ H ₄₃ SO ₁₁ : 647.

 Found: 647.

IR Spectrum (KBr): 3474, 1746, 1724, 1712, 1270, 1240, 1070 cm ^-1 .

¹ H-NMR (CDCl ₃ , 300 MHz), δ: 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), 4.83 (broad square, J = 5, 1 Hz, 1H), 4.66 (AVq, 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), 2.79 (m, 1H), 2.27 (s, 3H), 2.25 (m, 2H), 2.19 (s, 3H), 2.16 (s, 3H), 2.10 (s, 4H), 1.81 (m, 1H), 1.72 (s, 3H), 1.61 (m, 2H), 1, 16 (s, 3H); 1.03 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ: 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.

К раствору гексаметилдисилазана (0,40 мл, 1,90 ммоль) в 15 мл тетрагидрофурана добавляли раствор н-бутиллития (0,75 мл, 2,5 М в гексанах, 1,88 ммоль) и перемешивали 5 минут при -55^oC. К этому раствору добавляли 7-метилтиометилбаккатин III (соединение примера 6, 1,03 г, 1,59 ммоль) в 10 мл тетрагидрофурана и перемешивали 10 минут до добавления 10 мл раствора (3R, 4R)-1-(трет-бутилоксикарбонил)-4-(2-фурил)- 3-(триэтилсилилокси)-2-азетидинона (883 мг, 2,40 ммоль). Убирали холодную ванну, и заменяли ее ванной при 0^oC, после чего реакционную смесь перемешивали 30 минут. Раствор разбавляли этилацетатом и промывали насыщенным раствором NH₄Cl, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (2,5:1, гексан:этилацетат) и получили 1,5 г продукта реакции сочетания 3'-N-дебензоил-3'-дезфенил-3'-N-(трет-бутилоксикарбонил)-3'- (2-фурил)-7-O-метилтиометил-2'-O-триэтилсилилпаклитаксела (93%).Example 7. The triethanolamine salt of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -2'-O-ethyloxycarbonyl-7-O-phosphonoxymethylpalitaxel
(a) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -7-O-methylthiomethylpaclitaxel

To a solution of hexamethyldisilazane (0.40 ml, 1.90 mmol) in 15 ml of tetrahydrofuran was added a solution of n-butyllithium (0.75 ml, 2.5 M in hexanes, 1.88 mmol) and stirred for 5 minutes at -55 ^° C To this solution was added 7-methylthiomethylbaccatin III (compound of Example 6, 1.03 g, 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- (triethylsilyloxy) -2-azetidinone (883 mg, 2.40 mmol). The cold bath was removed and replaced with a bath at 0 ^° C., after which the reaction mixture was stirred for 30 minutes. The solution was diluted with ethyl acetate and washed with saturated NH ₄ Cl solution, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (2.5: 1, hexane: ethyl acetate) to obtain 1.5 g of the reaction product of the combination of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3'- (2-furyl) -7-O-methylthiomethyl-2'-O-triethylsilylpaclitaxel (93%).

Mass Spectrum FAB / NOBA / [M + Na] Calc. for C ₅₀ H ₇₁ NSSiO ₁₆ : 1036, found: 1036.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.07 (d, J = 7.2 Hz, 2H), 7.56 (m, 1H), 7.46 (t, J = 7.5 Hz , 2H), 7.36 (m, 1H), 6.56 (s, 1H), 6.33 (m, 1H), 6.20 (m, 2H), 5.67 (d, J = 6, 9 Hz, 1H), 5.29 (bs, 2H), 4.94 (d, J = 7.8 Hz, 1H), 4.75 (s, 1H), 4.65 (s, 2H), 4 28 (m, 2H), 4.16 (d, J = 8.1 Hz, 1H), 3.89 (d, J = 6.9 Hz, 1H), 2.80 (m, 1H), 2 46 (s, 3H), 2.37 (m, 1H), 2.22 (m, 1H), 2.16 (s, 3H), 2.10 (s, 3H), 2.04 (s, 3H), 1.84 (m, 1H), 1.74 (s, 3H), 1.65 (m, 1H), 1.33 (s, 9H), 1.20 (s, 3H), 1, 19 (s, 3H), 0.81 (t, J = 7.8 Hz, 9H), 0.47 (m, 6H).

¹³ C NMR (CDCl ₃ ; 75.5 Hz), δ: 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 a solution of the 2'-triethylsilyl ether obtained above (330 mg, 0.32 mmol) in 7 ml of tetrahydrofuran was added tetrabutylammonium fluoride (0.35 ml, 1.0 M THF, 0.35 mmol) and stirred for 10 minutes. The solution was diluted with ethyl acetate and washed with brine, dried over MgSO ₄ and concentrated, and the residue was chromatographed on silica gel (2: 1, hexane: ethyl acetate) to give 301 mg of the title compound (95%).

Mass spectrum FAB (NOBA) M + H calculated for C ₄₅ H ₅₈ NO ₁₆ S: 900. Found: 900.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.07 (d, J = 7.3 Hz, 2H), 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), 5.29 (m, 2H ), 4.92 (d, J = 8.0 Hz, 1H), 4.70 (m, 1H), 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), 2.38 (s, 3H), 2.32 (m, 2H), 2.16 (s, 3H), 2.10 (s, 3H), 2.02 (s , 3H), 1.77 (m, 3H) 1.73 (s, 3H), 1.33 (s, 9H), 1.17 (s, 3H), 1.12 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ 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.

К раствору полученного на стадии (a) продукта (864 мг, 0,96 ммоль) в 50 мл дихлорметана при 0^oC добавляли диизопропилэтиламин (2,0 мл, 11,5 ммоль) и этилхлорформиат (0,50 мл, 5,25 ммоль) и перемешивали 4 часа. Раствор разбавляли дихлорметаном и промывали насыщенным раствором бикарбонат, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (1:1, гексан:этилацетат) и получили 884 мг 2'-этилкарбоната указанного в заголовке соединения (95%).(b) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -2'-O-ethyloxycarbonyl-7-O-methylthiomethylpaclitaxel

To a solution of the product obtained in stage (a) (864 mg, 0.96 mmol) in 50 ml of dichloromethane at 0 ^° C. were added diisopropylethylamine (2.0 ml, 11.5 mmol) and ethyl chloroformate (0.50 ml, 5.25 mmol) and stirred for 4 hours. The solution was diluted with dichloromethane and washed with saturated bicarbonate solution, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (1: 1, hexane: ethyl acetate) to obtain 884 mg of 2'-ethyl carbonate of the title compound (95%).

Mass spectrum FAB (NOBA) M + H calculated for C ₄₈ H ₆₂ NO ₁₈ S: 972.3688. Found: 972.3654.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.09 (d, J = 7.8 Hz, 2H), 7.57 (t, J = 7.5 Hz, 1H), 7.46 (t , J = 7.8 Hz, 2H), 7.38 (s, 1H), 6.55 (s, 1H), 6.35 (m, 1H), 6.27 (m, 1H), 6.22 (t, J = 7.8 Hz, 1H), 5.67 (d, J = 7.2 Hz, 1H), 5.51 (d, J = 9.9 Hz, 1H), 5.34 (d , J = 2.4 Hz, 1H), 5.25 (d, J = 10.2 Hz, 1H), 4.95 (d, J = 8.1 Hz, 1H), 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), 2.36-2.21 (m, 2H), 2.16 (s, 3H), 2.11 (s, 3H), 2.09 (s, 3H), 1.83 (m, 1H), 1.74 (s, 3H), 1.67 (s, 1H), 1.59 (s, 1H), 1.34 (s, 9H), 1.29 (t, J = 7.2 Hz, 3H), 1.20 (s, 3H), 1.18 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ: 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.

К раствору полученного на стадии (b) продукта (230 мг, 0,236 ммоль) в 10 мл безводного тетрагидрофурана добавляли 300 мг

молекулярных сит, дибензилфосфат (270 мг, 0,98 ммоль) и перекристаллизованный NIS (62 мг, 0,28 ммоль). К этому раствору добавляли трифторметансульфонат серебра (45 мг, 0,17 ммоль), и раствор перемешивали 3 часа. Раствор фильтровали через целит и разбавляли этилацетатом, промывали 10%-ным раствором NaS₂O₈, насыщенным раствором бикарбоната и рассолом, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (15% ацетонитрила в хлороформе) и получили 219 мг дибензилфосфата указанного в заголовке соединения (77%).(c) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) - 2 -'-O-ethyloxycarbonyl-7-O-dibenzylphosphonoxymethylpaclitaxel

To a solution of the product obtained in stage (b) (230 mg, 0.236 mmol) in 10 ml of anhydrous tetrahydrofuran was added 300 mg

molecular sieves, dibenzyl phosphate (270 mg, 0.98 mmol) and recrystallized NIS (62 mg, 0.28 mmol). Silver trifluoromethanesulfonate (45 mg, 0.17 mmol) was added to this solution, and the solution was stirred for 3 hours. The solution was filtered through celite and diluted with ethyl acetate, washed with 10% NaS ₂ O ₈ solution, saturated bicarbonate solution and brine, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (15% acetonitrile in chloroform) to give 219 mg of the dibenzyl phosphate of the title compound (77%).

Mass Spectrum FAB (NOBA) Calc. for C ₆₁ H ₇₂ NPO ₂₂ Na: 1224. Found: 1224.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 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), 6.35 (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 (d, J = 9.9 Hz, 1H), 5.01 (dd, J = 8.1 and 6.3 Hz, 5H), 4.86 (d, J = 8.4 Hz, 1H), 4.29 - 4.09 (m, 4H), 3.85 (d, J = 6.9 Hz, 1H), 2.77 (m, 1H), 2.40 (s, 3H), 2.30 (m, 2H), 2.16 (s, 3H), 1.99 (s, 3H), 1.94 (m, 1H), 1.70 (s, 3H), 1.67 (s, 1H) , 1.54 (s, 1H), 1.34 (s, 9H), 1.28 (t, J = 7.2 Hz, 3H), 1.20 (s, 3H), 1.17 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ: 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.

К раствору полученного на стадии (c) продукта (311 мг, 0,259 ммоль) в 25 мл этилацетата добавляли 60 мг палладия на угле (10%), и раствор перемешивали в среде водорода в течение 30 минут. Удаляли катализатор путем фильтрования через целит, и фильтрат концентрировали в вакууме. Остаток растворяли в 3 мл этилацетата, и добавляли триэтаноламин (2,3 мл, 0,1 М в этилацетоне, 0,23 ммоль). Раствор концентрировали и остаток хроматографировали на C18 (40% ацетонитрила в воде) и лиофилизировали, получив в результате 205 мг триэтаноламиновой соли (67%) фосфоновой кислоты.(d) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3'- (2-furyl) -2'-O-ethyloxycarbonyl-7-O-phosphonooxymethylpaclitaxel triethanolamine salt

To a solution of the product obtained in step (c) (311 mg, 0.259 mmol) in 25 ml of ethyl acetate was added 60 mg of palladium on carbon (10%), and the solution was stirred in hydrogen medium for 30 minutes. The catalyst was removed by filtration through celite, and the filtrate was concentrated in vacuo. The residue was dissolved in 3 ml of ethyl acetate, and triethanolamine (2.3 ml, 0.1 M in ethyl acetate, 0.23 mmol) was added. The solution was concentrated and the residue was chromatographed on C18 (40% acetonitrile in water) and lyophilized, resulting in 205 mg of triethanolamine salt (67%) of phosphonic acid.

Mass spectrum FAB (NOBA): M + Na calc. for C ₄₇ H ₆₀ HPO ₂₂ Na: 1044. Found: 1044.

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

¹ H NMR (d ₆ acetone / D ₂ O, 300 MHz), δ: 8.09 (d, J = 7.2 Hz, 2H), 7.62 (m, 2H), 7.52 (t, J = 7.5 Hz, 2H), 6.48 (d, J = 3.3 Hz, 1H), 6.42 (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), 5.30 (d, J = 3.6 Hz, 1H), 5, 17 (br, 1H), 5.01 (br, J = 9.0 Hz, 1H), 4.19 (br, 1H), 4.18 (m, 5H), 3.95 (m, 6H), 3.87 (d, J = 6.9 Hz, 1H), 3.68 (s, 7H), 3.50 (pcs, J = 4.8 Hz, 6H), 2.95 (m, 1H), 2.44 (s, 3H), 2.41 (m, 2H), 2.16 (s, 3H), 1.9 (s, 3H), 1.94 (m, 1H), 1.68 (s , 3H), 1.34 (s, 9H), 1.24 (t, J = 6.9 Hz, 3H), 1.17 (s, 6H).

К раствору ГМДС (0,5 мл, 2,4 ммоль) в 18 мл ТГФ при -55^oC добавляли н-бутиллитий (0,85 мл, 2,5 М в гексанах, 2,1 ммоль). Через 10 минут добавляли по каплям 7-метилтиометилбаккатин III (1,15 г, 1,78 ммоль) в 18 мл ТГФ и перемешивали на холоду в течение 10 минут. Добавляли (±)цис-1- (трет-бутилоксикарбонил)-4-(2-тиенил)-3-(триэтилсилилокси)- 2-азетидинон (2,80 г, 7,3 ммоль) в 18 мл ТГФ, и позволяли холодной ванне медленно нагреваться до 0^oC в течение 30 минут. Раствор разбавляли этилацетатом, промывали насыщенным раствором NH₄Cl, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (5 : 1, гексан : этилацетат), получив 1,87 г рекуперированного лактамами (3 : 1, гексан : этилацетат), что дало 1,44 г продукта реакции сочетания 3'-N-дебензоил-3'-дезфенил-3'-N-(трет-бутилоксикарбонил)-3'- (2-тиенил)-7-O-метилтиометил-2'-O-триэтилсилилпаклитаксела (78%).Example 8. The triethanolamine salt of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3'- (2-thienyl) -2'-O-ethyloxycarbonyl-7-O-phosphonoxymethylpaclitaxel
(a) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -7-O-methylthiomethylpaclitaxel

To a solution of HMDS (0.5 ml, 2.4 mmol) in 18 ml of THF at -55 ^° C was added n-butyllithium (0.85 ml, 2.5 M in hexanes, 2.1 mmol). After 10 minutes, 7-methylthiomethylbaccatin III (1.15 g, 1.78 mmol) in 18 ml of THF was added dropwise and stirred in the cold for 10 minutes. (±) cis-1- (tert-butyloxycarbonyl) -4- (2-thienyl) -3- (triethylsilyloxy) -2-azetidinone (2.80 g, 7.3 mmol) in 18 ml of THF was added and allowed to cool the bath is slowly heated to 0 ^o C for 30 minutes. The solution was diluted with ethyl acetate, washed with saturated NH ₄ Cl solution, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (5: 1, hexane: ethyl acetate) to obtain 1.87 g of recovered lactams (3: 1, hexane: ethyl acetate), which afforded 1.44 g of the reaction product of the 3'-N-debenzoyl-3'- combination desphenyl-3'-N- (tert-butyloxycarbonyl) -3'- (2-thienyl) -7-O-methylthiomethyl-2'-O-triethylsilylpaclitaxel (78%).

Mass spectrum FAB (NOBA): M + Na calc. for C ₅₁ H ₇₁ NO ₁₅ S ₂ SiNa: 1052. Found: 1052.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.09 (d, J = 7.2 Hz, 2H), 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), 5.68 (d, J = 6.9 Hz, 1H), 5.49 (br, 1H), 5.39 (br, J = 9.6 Hz, 1H), 4.94 (d , J = 7.8 Hz, 1H), 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), 2.37 (m, 1H), 2.20 (m, 1H), 2.17 (s, 3H), 2.10 (s, 3H), 2.03 (s, 3H), 1.84 (m, 1H), 1.74 (s, 3H) 1.68 (s, 1H), 1.62 (s, 1H), 1.31 (s, 9H), 1.20 (s, 6H), 0.84 (t, J = 7.8 Hz, 9H), 0.50 (m, 6H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ: 201.9, 171.1, 170.7, 170.1, 169.3, 167.0, 155.1 142.8, 140.9, 133.6, 132.9, 130.2, 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 a solution of the 2'-triethylsilyl ether obtained above (1.41 g, 1.37 mmol) in 14 ml of tetrahydrofuran was added tetrabutylammonium fluoride (1.4 ml, 1.0 M in THF, 1.40 mmol). The solution was stirred for 30 minutes, diluted with ethyl acetate, washed with brine, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (hexane: ethyl acetate, 1: 1) to give 1.16 g of the title compound (92%).

Mass spectrum FAB (NOBA): M + Na calc. for C ₄₅ H ₅₇ NO ₁₅ S ₂ Na: 938. Found: 938.

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

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.08 (d, J = 7.2 Hz, 2H), 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), 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 (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), 2.36 (s, 3H), 2.34 (, 2H), 2.17 (s, 3H), 2.10 (s, 3H), 2.00 (s, 3H), 1, 83 (m, 1H), 1.74 (s, 3H), 1.72 (s, 1H), 1.61 (s, 1H), 1.33 (s, 9H), 1.21 (s, 3H ), 1.18 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 MHz), δ: 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, 47.2, 43.2, 35.3, 32.9, 28.2, 26.4 , 22.6, 20.9, 15.1, 14.7, 10.8.

К раствору полученного на стадии (a) продукта (621 мг, 0,677 ммоль) в 35 мл дихлорметана при 0^oC добавляли диизопропилэтиламин (1,20 мл, 6,89 ммоль) и этилхлорформиат (0,35 мл, 3,7 ммоль) и перемешивали 1 час. Убирали холодную ванну, и раствор перемешивали 2 часа, разбавляли дихлорметаном, промывали насыщенным раствором бикарбоната, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (гексан : этилацетат = 1 : 1) и получили 528 мг указанного в заголовке соединения (79%).(b) Preparation of 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -2'-O-ethyloxycarbonyl-7-O-methylthiomethylpaclitaxel

To a solution of the product obtained in stage (a) (621 mg, 0.677 mmol) in 35 ml of dichloromethane at 0 ^° C. were added diisopropylethylamine (1.20 ml, 6.89 mmol) and ethyl chloroformate (0.35 ml, 3.7 mmol) and stirred for 1 hour. The cold bath was removed and the solution was stirred for 2 hours, diluted with dichloromethane, washed with saturated bicarbonate solution, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (hexane: ethyl acetate = 1: 1) to give 528 mg of the title compound (79%).

Mass spectrum FAB (NOBA): M + Na calc. for C ₄₃ H ₆₁ NO ₁₇ S ₂ Na: 1010. Found: 1010.

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

¹ H-NMR (CDCl ₃ , 300 MHz), δ: 8.09 (d, J = 7.2 Hz, 2H), 7.58 (m, 1H), 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), 5.68 (d, J = 6.9 Hz, 2H), 5.33 (d, J = 9.9 Hz, 1H), 5.25 (d, J = 2.4 Hz, 1H), 4.94 (d , J = 7.8 Hz, 1H), 4.65 (s, 2H), 4.33 - 4.08 (m, 5H), 3.88 (d, J = 6.9 Hz, 1H), 2 80 (m, 1H), 2.40 (s, 3H), 2.40 - 2.20 (m, 2H), 2.16 (s, 3H), 2.11 (s, 3H), 2, 07 (s, 3H), 1.83 (m, 1H), 1.74 (s, 3H), 1.69 (s, 1H), 1.60 (s, 1H), 1.33 (s, 9H ), 1.31 (t, J = 7.2 Hz, 3H), 1.20 (s, 3H), 1.19 (s, 3H).

¹³ C-NMR (CDCl ₃ , 75.5 Hz), δ: 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.

К раствору полученного на стадии (b) продукта (516 мг, 0,522 ммоль) в 15 мл безводного тетрагидрофурана добавляли 530 мг молекулярных сит

дибензилфосфат (576 мг, 2,09 ммоль) и перекристаллизованный N-иодсукцинимид (136 мг, 0,604 ммоль). К этому раствору добавляли трифторметансульфонат серебра (50 мг, 0,194 ммоль), и раствор перемешивали 1 час. Раствор фильтровали через целит, разбавляли этилацетатом, промывали 10%-ным раствором NaS₂O₈, насыщенным раствором бикарбоната и рассолом, высушивали над MgSO₄ и концентрировали. Остаток хроматографировали на силикагеле (15% ацетонитрила в хлороформе) и получили 535 мг указанного в заголовке соединения (84%).(c) Preparation of 3'-N- (debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -2'-O-ethyloxycarbonyl-7-O-dibenzylphosphonoxymethylpaclitaxel

To a solution of the product obtained in stage (b) (516 mg, 0.522 mmol) in 15 ml of anhydrous tetrahydrofuran, 530 mg of molecular sieves were added

dibenzyl phosphate (576 mg, 2.09 mmol) and recrystallized N-iodosuccinimide (136 mg, 0.604 mmol). Silver trifluoromethanesulfonate (50 mg, 0.194 mmol) was added to this solution, and the solution was stirred for 1 hour. The solution was filtered through celite, diluted with ethyl acetate, washed with 10% NaS ₂ O ₈ solution, saturated bicarbonate solution and brine, dried over MgSO ₄ and concentrated. The residue was chromatographed on silica gel (15% acetonitrile in chloroform) to give 535 mg of the title compound (84%).

Mass spectrum FAB (NOBA): M + Na calc. for C ₁₆ H ₇₂ NO ₂₁ PSNa: 1240. Found: 1240.

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

¹ H-NMR (CDCl ₃ , 300 MHz), δ: 8.08 (d, J = 7.0 Hz, 2H), 7.58 (m, 1H), 7.47 (t, J = 7.5 Hz, 2H), 7.28 (m, 11H), 6.99 (m, 2H), 6.33 (s, 1H), 6.22 (t, J = 7.8 Hz, 1H), 5, 66 (m, 2H), 5.39 (t, J = 6.6 Hz, 1H), 5.34 (d, J = 12 Hz, 1H), 5.22 (d, J = 2.4 Hz, 1H), 5.01 (dd, J = 8.1 and 6.0 Hz, 5H), 4.86 (d, J = 7.8 Hz, 1H), 4.29 - 4.08 (m, 5H ), 3.85 (d, J = 6.6 Hz, 1H), 2.76 (m, 1H), 2.39 (s, 3H), 2.35 - 2.18 (m, 2H), 2 16 (s, 3H) 1.97 (s, 4H), 1.69 (s, 4H), 1.33 (s, 9H), 1.30 (t, J = 7.2 Hz, 3H), 1.20 (s, 3H); 1.17 (s, 3H).

¹³ C NMR (CDCl ₃ , 75.5 Hz), δ: 197.4, 165.4, 164.9, 163.3, 162.7, 150.6, 149.7, 136.7, 136.0 , 129.4, 128.6, 125.9, 124.7, 124.3, 124.2, 124.1, 123.6, 122.9, 121.1, 121.0, 89.4, 79 , 8, 77.3, 76.5, 76.3, 74.4, 72.0, 70.7, 70.3, 67.7, 64.9, 64.9, 60.9, 52.7 , 46.5, 42.3, 38.9, 30.7, 23.8, 22.0, 18.3, 17.0, 16.4, 10.3, 9.8, 6.2.

К раствору полученного на стадии (c) продукта (512 мг, 0,42 ммоль) в 30 мл этилацетата добавляли 53 мг палладия на угле (10%), и раствор перемешивали в среде водорода в течение 3 часов. Удаляли катализатор путем фильтрования через целит, и фильтрат концентрировали в вакууме. Остаток растворяли в 2 мл этилацетата, и добавляли триэтаноламин (4,0 мл, 0,1 М этилацетате, 0,40 ммоль). Раствор концентрировали, и остаток хроматографировали на C18 (40% ацетонитрила в воде) и лиофилизировали, получив в результате 280 мг триэтаноламиновой соли (56%). Анализ путем HPLC показал чистоту соли, равную 96%.(d) Preparation of the 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -2'-O-ethyloxycarbonyl-7-O-phosphonooxymethyl paclitaxel triethanolamine salt

To a solution of the product obtained in step (c) (512 mg, 0.42 mmol) in 30 ml of ethyl acetate was added 53 mg of palladium on carbon (10%), and the solution was stirred in hydrogen medium for 3 hours. The catalyst was removed by filtration through celite, and the filtrate was concentrated in vacuo. The residue was dissolved in 2 ml of ethyl acetate, and triethanolamine (4.0 ml, 0.1 M ethyl acetate, 0.40 mmol) was added. The solution was concentrated, and the residue was chromatographed on C18 (40% acetonitrile in water) and lyophilized, resulting in 280 mg of triethanolamine salt (56%). HPLC analysis showed a salt purity of 96%.

Mass spectrum FAB (NOBA): M + Na calc. for C ₄₇ H ₆₀ NO ₂₁ PS: 1060. Found: 1060.

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

¹ H NMR (d ₆ acetone / D ₂ O, 300 MHz), δ: 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), 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), 4.86 (dd, J = 12.0 and 6.3 Hz, 1H), 4.17 (m, 5H), 4.00 (s, 7H), 3.92 (t, J = 4.8 Hz, 6H), 3.84 (d, J = 6.9 Hz, 1H), 3.48 (t, J = 5.4 Hz, 6H) 2.94 (m, 1H), 2.42 (s, 3H), 2.36 (m, 1H), 2.27 (m, 1H), 2.15 (s, 3H), 1.95 ( s, 4H), 1.66 (s, 3H), 1.30 (s, 9H), 1.23 (t, J = 7.2 Hz, 3H) 1.14 (s, 6H).

В сухую колбу в среде аргона, содержащую 7-O-триэтилсилил-10- дезацетилбаккатин III (2,093 г, 3,177 ммоль), добавляли сухой тетрагидрофуран (30 мл) и охлаждали до -70^oC. Туда же добавляли по каплям 1,6 M н-бутиллитий (2,38 мл, 3,81 ммоль). После перемешивания в течение 15 минут добавляли по каплям бензилхлорформиат (0,91 мл, 6,35 ммоль). Полученную смесь перемешивали 3 часа с постепенным нагреванием до окружающей температуры. Реакцию быстро прекращали посредством 25 мл насыщенного раствора NH₄Cl, смесь промывали рассолом и высушивали над MgSO₄. Флеш-хроматография (силикагель, 30 - 45% этилацетата в гексане) дала 2,24 г (89%) указанного в заголовке соединения в виде белой пены.Example 9. 10-desacetyl-3'-N-desbenzoyl-3'-N- (tert-butyloxycarbonyl) -10-O- (phosphonooxymethyl) paclitaxel
(a) Preparation of 10-deacetyl-10-O-benzyloxycarbonyl-7-O-triethylsilyl baccatin III

Dry tetrahydrofuran (30 ml) was added to a dry flask in argon medium containing 7-O-triethylsilyl-10-deacetylbaccatin III (2.093 g, 3.177 mmol) and cooled to -70 ^° C. 1.6 M was added dropwise thereto. n-butyllithium (2.38 ml, 3.81 mmol). After stirring for 15 minutes, benzyl chloroformate (0.91 ml, 6.35 mmol) was added dropwise. 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 NH ₄ Cl, the mixture was washed with brine and dried over MgSO ₄ . Flash chromatography (silica gel, 30-45% ethyl acetate in hexane) gave 2.24 g (89%) of the title compound as a white foam.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 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), 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), 2.29 (m, 4H), 2.20 (s, 3H), 2.03 (d, J = 5, 0 Hz, 1H), 1.92 - 1.83 (m, 1H), 1.68 (s, 3H), 1.17 (s, 3H), 1.04 (s, 3H), 0.91 ( t, J = 7.5 Hz, 9H), 0.57 (q, J = 7.4 Hz, 6H).

В сухую колбу, содержащую полученный на стадии (a) продукт (3,50 г, 4,42 ммоль), добавляли небольшое количество толуола, после чего раствор концентрировали под вакуумом. Эту колбу помещали в среду аргона, и добавляли в нее 100 мл сухого тетрагидрофурана. Колбу охлаждали до -70^oC, и добавляли в нее по каплям 1,0 М гексаметилдисилазид лития (6,19 мл, 6,19 ммоль). После перемешивания в течение 20 минут добавляли по каплям раствор (3R,4S)-1-(трет-бутилоксикарбонил)-4-фенил-3-триэтилсилилокси-2- азетидинона (2,58 г, 7,07 ммоль) в 10 мл сухого ТГФ. Реакционную смесь перемешивали 3,5 часов, постепенно нагревая до температуры окружающей среды. Затем ее быстро охлаждали 70 мл насыщенного раствора NH₄Cl, промывали рассолом и высушивали над MgSO₄. Флеш-хроматография (силикагель, 5 - 15% этилацетата в гексанах) дала 5,12 г (99%) указанного в заголовке соединения в виде белой пены.(b) Preparation of 10-deacetyl-10-O-benzyloxycarbonyl-3'-N-desbenzoyl-3'-N- (tert-butyloxycarbonyl) -2'-7-bis-O- triethylsilylpaclitaxel

A small amount of toluene was added to a dry flask containing the product obtained in stage (a) (3.50 g, 4.42 mmol), after which the solution was concentrated in vacuo. This flask was placed in argon medium, and 100 ml of dry tetrahydrofuran was added to it. The flask was cooled to -70 ^° C, and 1.0 M lithium hexamethyldisilazide (6.19 ml, 6.19 mmol) was added dropwise thereto. After stirring for 20 minutes, a solution of (3R, 4S) -1- (tert-butyloxycarbonyl) -4-phenyl-3-triethylsilyloxy-2-azetidinone (2.58 g, 7.07 mmol) in 10 ml dry was added dropwise. THF The reaction mixture was stirred for 3.5 hours, gradually warming to ambient temperature. Then it was quickly cooled with 70 ml of a saturated solution of NH ₄ Cl, washed with brine and dried over MgSO ₄ . Flash chromatography (silica gel, 5-15% ethyl acetate in hexanes) gave 5.12 g (99%) of the title compound as a white foam.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.11 (d, J = 8.0 Hz, 2H), 7.60 - 7.58 (m, 1H), 7.48 (t, J = 8.0 Hz, 2H), 7.24 - 7.26 (m, 10H), 6.32 - 6.26, (m, 2H), 5.69 (d, J = 7 Hz, 1H), 5 47 (br, 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), 4.31 (d, J = 8.3 Hz, 1H), 4.17 (d, = 8.3 Hz , 1H), 3.81 (d, J = 7.0 Hz, 1H), 2.53 (s, 3H), 2.48 - 2.33 (m, 1H), 2.22 - 2.17 ( m, 1H), 2.09 (s, 3H), 1.95 - 1.86 (m, 1H), 1.70 - (s, 3H), 1.65 (s, 1H), 1.52 ( s, 1H), 1.30 (s, 9H), 1.26 - 1.19 (m, 6H), 0.94 - 0.87 (m, 9H), 0.80 - 0.75 (m, 9H), 0.61-0.53 (m, 6H), 0.48-0.30 (m, 6H).

Полученный на стадии (b) продукт (5,12 г, 4,40 ммоль) растворяли в 100 мл этилацетата, переносили в сосуд Парра и помещали под защитный слой аргона. Туда добавляли 10% палладия на угле (2,4 г), и реакционную смесь помещали на гидрогенизационную установку Парра (55 фунтов на кв. дюйм, т.е. 3,87 кг/см²) на 8 часов. Реакционную смесь фильтровали через целит и концентрировали. Флеш-хроматография (силикагель, 15 - 20% этилацетата в гексане) дала 3,24 г (79%) указанного в заголовке соединения в виде белой пены. Гидролиз 2'-триэтилсилильной группы продукта, полученного на стадии Парра.(c) Preparation of 10-deacetyl-3'-N-debenzoyl-3'-N- (tert-butyloxycarbonyl) -7-O-triethylsilylpaclitaxel

The product obtained in step (b) (5.12 g, 4.40 mmol) was dissolved in 100 ml of ethyl acetate, transferred to a Parr vessel and placed under a protective argon layer. 10% palladium-carbon (2.4 g) was added thereto, and the reaction mixture was placed on a Parr hydrogenation unit (55 psi, i.e. 3.87 kg / cm ² ) for 8 hours. The reaction mixture was filtered through celite and concentrated. Flash chromatography (silica gel, 15-20% ethyl acetate in hexane) gave 3.24 g (79%) of the title compound as a white foam. Hydrolysis of the 2'-triethylsilyl group of the product obtained in the Parr step.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 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 - 6.17 (m, 1H), 5.64 (d, J = 7.2 Hz), 5, 42 (d, J = 9.4 Hz, 1H), 5.28 - 5.25 (m, 1H), 5.12 (s, 1H), 4.92 (d, J = 8.6 Hz, 1H ), 4.62 (bs, 1H), 4.38-4.28 (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 - 2.40 (m, 1H), 2.36 (s, 3H), 2.25 (br, J = 8.7 Hz, 2H), 1.99 - 1.91 (m, 1H), 1.85 (s, 3H), 1.74 (s, 3H), 1.69 (s, 1H), 1, 67 (s, 1H), 1.35 (s, 9H), 1.22 (s, 3H), 1.11 (s, 3H), 0.93 (t, J = 7.5 Hz, 9H), 0.61 - 0.49 (m, 6H).

В колбу, содержащую полученный на стадии (с) продукт (3,24 г, 3,51 ммоль), добавляли 30 мл сухого дихлорметана. Колбу помещали под аргон и охлаждали до 0^oC. К реакционной смеси добавляли N,N-диизопропилэтиламин (1,22 мл, 7,02 ммоль), а затем добавляли по каплям бензилхлорформиат (1,00 мл, 7,02 ммоль). Через 15 минут охлажденную ванну убирали, и позволяли реакционной смеси перемешиваться при температуре окружающей среды в течение 7 часов. Затем смесь резко охлаждали посредством 30 мл насыщенного раствора NH₄Cl, промывали рассолом и высушивали над MgSO₄. Флеш-хроматография (силикагель, 7 - 20% этилацетата в гексане) дала 3,24 г (89%) указанного в заголовке соединения в виде белого твердого вещества.(d) Preparation of 10-deacetyl-2'-O-benzyloxycarbonyl-3'-N-debenzoyl-3'-N- (tert-butyloxycarbonyl) -7-O-triethylsilylpaclitaxel

To a flask containing the product obtained in step (c) (3.24 g, 3.51 mmol) was added 30 ml of dry dichloromethane. The flask was placed under argon and cooled to 0 ^° C. N, N-diisopropylethylamine (1.22 ml, 7.02 mmol) was added to the reaction mixture, and then benzyl chloroformate (1.00 ml, 7.02 mmol) was added dropwise. After 15 minutes, the cooled bath was removed and the reaction mixture was allowed to mix at ambient temperature for 7 hours. The mixture was then quenched with 30 ml of a saturated solution of NH ₄ Cl, washed with brine and dried over MgSO ₄ . Flash chromatography (silica gel, 7-20% ethyl acetate in hexane) gave 3.24 g (89%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.10 (d, J = 8.0 Hz, 2H), 7.62 - 7.57 (m, 1H), 7.48 (t, J = 8.0 Hz, 2H), 7.40 - 7.26 (m, 10H), 6.33 - 6.27 (m, 1H), 5.66 (d, J = 7.0 Hz, 1H), 5.49 - 5.42 (m, 2H), 5.31 (s, 1H), 5.22 - 5.13 (m, 3H), 4.93 (d, J = 9.4 Hz, 1H) , 4.38 (dd, J = 6.5 and 10.7 Hz, 1H), 4.34 - 4.28 (m, 2H), 4.18 (d, J = 8.3 Hz, 1H), 3.90 (d, J = 6.7 Hz, 1H), 2.52 - 2.30 (m, 4H), 2.24 - 2.20 (m, 1H), 1.97 - 1.87 ( m, 3H), 1.74 (s, 3H), 1.59 (s, 3H), 1.32 (s, 9H), 1.26 (s, 3H), 1.11 (s, 3H), 0.96-0.88 (m, 9H); 0.61-0.48 (m, 6H).

Полученный на стадии (d) продукт растворяли в 13,5 мл (54%) ДМСО, 8,75 мл (35%) уксусного ангидрида и 2,75 мл (11%) ледяной уксусной кислоты и помещали в атмосферу аргона. Реакционную смесь перемешивали 56 часов, после чего ее разбавляли этилацетатом до объема 60 мл. Раствор промывали насыщенным раствором NaHCO₃, пока он не стал нейтральным при проверке индикаторной бумаги для определения pH, и затем промывали рассолом. Органическую фракцию высушивали MgSO₄ и концентрировали. Флеш-хроматография (15 - 20% этилацетата в гексане) дала 3,12 г неочищенной белой пены с требуемым тиометилацетальным продуктом (т. е. 10-дезацетил-2'-O-бензилоксикарбонил-3'-N-дебензоил- 3'-N-(трет-бутилоксикарбонил)-10-O-(метилтиометил)-7-O- триэтилсилилпаклитакселом), составляющим 70% материала по ЯМР.(e) Preparation of 10-deacetyl-2'-O-benzyloxycarbonyl-3'-N-debenzoyl-3'-N- (tert-butyloxycarbonyl) -10-O- (dibenzylphosphonooxymethyl) -7-O- triethylsilyl paclitaxel

The product obtained in step (d) was dissolved in 13.5 ml (54%) of DMSO, 8.75 ml (35%) of acetic anhydride and 2.75 ml (11%) of glacial acetic acid and placed in an argon atmosphere. The reaction mixture was stirred for 56 hours, after which it was diluted with ethyl acetate to a volume of 60 ml. The solution was washed with saturated NaHCO ₃ solution until it became neutral when checking pH indicator paper, and then washed with brine. The organic layer was dried with MgSO ₄ and concentrated. Flash chromatography (15-20% ethyl acetate in hexane) gave 3.12 g of a crude white foam with the desired thiomethyl acetal product (i.e. 10-deacetyl-2'-O-benzyloxycarbonyl-3'-N-debenzoyl-3'- N- (tert-butyloxycarbonyl) -10-O- (methylthiomethyl) -7-O-triethylsilylpaclitaxel), constituting 70% of the NMR material.

(3,12 г), и полученную гетерогенную смесь интенсивно перемешивали. Добавляли к ней посредством канюли раствор перекристаллизованного N-иодсукцинимида (0,830 г, 3,69 ммоль) и дибензилфосфата (1,027 г, 3,69 ммоль) в сухом тетрагидрофуране (46 мл). Полученную смесь перемешивали 5 часов, фильтровали через слой целита и разбавляли до объема 250 мл этилацетата. Промывали ее холодным 2%-ным раствором NaHCO₃ (2 х 125 мл), холодным 6%-ным раствором NaHCO₃ (2 х 125 мл) и рассолом. Органическую фазу высушивали (MgSO₄) и концентрировали. Флеш-хроматография (силикагель 25 - 35% этилацетата в гексане) дала 1,52 г (40%) указанного в заголовке соединения в виде белого твердого вещества.Then, the above crude mixture (3.12 g) was dissolved in 1,2-dichloroethane (61 ml) and placed under a protective argon layer. Powdered molecular sieves were added.

(3.12 g), and the resulting heterogeneous mixture was intensively mixed. A solution of recrystallized N-iodosuccinimide (0.830 g, 3.69 mmol) and dibenzyl phosphate (1.027 g, 3.69 mmol) in dry tetrahydrofuran (46 ml) was added via cannula. The resulting mixture was stirred for 5 hours, filtered through a pad of celite and diluted to a volume of 250 ml of ethyl acetate. It was washed with cold 2% NaHCO ₃ solution (2 x 125 ml), cold 6% NaHCO ₃ solution (2 x 125 ml) and brine. The organic phase was dried (MgSO ₄ ) and concentrated. Flash chromatography (silica gel 25-35% ethyl acetate in hexane) gave 1.52 g (40%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.08 (d, J = 7.0 Hz, 2H), 7.59 - 7.55 (m, 1H), 7.46 (t, J = 7.2 Hz, 2H), 7.38 - 7.25 (m, 20H), 6.30 (t, J = 8.5 Hz, 1H), 5.65 (d, J = 6.8
Hz, 1H), 5.49-5.39 (m, 4H), 5.32 (s, 1H), 5.18-4.19 (m, 4H), 4.93 (d, J = 9, 2 Hz, 1H), 4.44 (dd, J = 6.6 and 10.2 Hz, 1H), 4.31 (d, J = 8.4 Hz, 1H), 4.16 (d, J = 8.5 Hz, 1H), 3.80 (d, J = 6.9 Hz, 1H), 2.69 - 2.39 (m, 4H), 2.33 - 2.23 (m, 3H), 2.03 (s, 3H), 1.90 (t, J = 12.6 Hz, 1H), 1.68 - 1.63 (m, 6H), 1.28 (s, 9H), 1.16 - 1.10 (m, 6H), 0.93 (t, J = 7.4 Hz, 9H), 0.55 (q, J = 7.8 Hz, 6H).

¹³ C NMR (CDCl ₃ , 75.5 MHz), δ: 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.

Раствор полученного на стадии (е) продукта (50,8 мг, 0,038 ммоль) в сухом тетрагидрофуране (2,5 мл) под аргоном охлаждали до -40^oC. К этому раствору добавляли по каплям тетрабутиламмонийфторид (0,057 мл, 0,057 ммоль) в тетрагидрофуране (1,0 М). Реакционную смесь перемешивали 1,5 часа при постепенном нагреве до -20^oC. Смесь резко охлаждали 15 мл насыщенного раствора NH₄Cl и разбавляли 30 мл этилацетата. Органическую фазу промывали NaHCO₃ (2 х 15 мл) и рассолом. Затем ее высушивали (MgSO₄) и концентрировали. Препаративная тонкослойная хроматография (силикагель, 50% этилацетата в гексане) дала 36 мг (77%) указанного в заголовке соединения в виде белого порошка.(f) Preparation of 10-deacetyl-2'-O-benzyloxycarbonyl-3'-N-debenzoyl-3'-N- (tert-butyloxycarbonyl) -10-O- (dibenzylphosphonooxymethyl) paclitaxel

A solution of the product obtained in step (e) (50.8 mg, 0.038 mmol) in dry tetrahydrofuran (2.5 ml) under argon was cooled to -40 ^° C. To this solution tetrabutylammonium fluoride (0.057 ml, 0.057 mmol) was added dropwise to tetrahydrofuran (1.0 M). The reaction mixture was stirred for 1.5 hours while gradually heating to -20 ^° C. The mixture was sharply cooled with 15 ml of a saturated solution of NH ₄ Cl and diluted with 30 ml of ethyl acetate. The organic phase was washed with NaHCO ₃ (2 x 15 ml) and brine. Then it was dried (MgSO ₄ ) and concentrated. Preparative thin layer chromatography (silica gel, 50% ethyl acetate in hexane) gave 36 mg (77%) of the title compound as a white powder.

¹ H NMR (CDCl ₃ ; 300 MHz), δ: 8.10 (d, J = 8.5 Hz, 2H), 7.60 - 7.55 (m, 1H), 7.49 - 7.44 ( m, 2H), 7.36 - 7.18 (m, 20H), 6.27 - 6.22 (m, 1H), 5.78 (s, 1H), 5.67 (d, J = 7, 0 Hz, 1H), 5.44 - 5.34 (m, 3H), 5.27 (d, J = 2.2 Hz, 1H), 5.24 - 5.05 (m, 4H), 5, 01 - 4.91 (m, 4H), 4.39 - 4.28 (m, 2H), 4.17 (d, J = 8.2 Hz, 1H), 3.87 (d, J = 7, 0 Hz, 1H), 2.58 - 2.51 (m, 1H), 2.41 (s, 3H), 2.40 - 2.18 (m, 2H), 2.00 - 1.87 (m 5H), 1.73-1.69 (m, 4H), 1.30 (s, 9H), 1.22-1.15 (m, 6H).

Mass spectrum (FAB): m / z ⁺ : 1231.

Колбу Парра емкостью 500 мл загружали 10-дезацетил-2'-O- бензилоксикарбонил-3'-N-дезбензоил-3'-N-(тетра-бутилоксикарбонил)-10-O- (дибензилфосфонооксиметил)паклитакселом (264,9 мг, 0,215 ммоль) и этилацетатом (20 мл). Потом колбу обдували струей аргона, и добавляли в нее 10% Pd/C (318 мг). Полученную смесь помещали на установку Парра в среду водорода под давлением 55 фунтов на кв. дюйм (3,86 кг/см²). Реакцию контролировали путем HPLC (70:30 буфер CH₃CN/Q8, pH 6,0, 1,00 мл/мин, колонка Зорбакса C-18, 25,0 см, λ = 230 нм), до тех пор, пока не стало видно исходного материала (12,5 часов). Смесь фильтровали через слой целита, который промывали этилацетатом и небольшим количеством дихлорметана. Полученный фильтрат концентрировали, и остаток растворяли в дихлорметане (5 мл). Добавление гексана вызвало образование осадка, из которого было выделено 140,3 мг свободной кислоты (чистота при определении путем HPLC - 80%) в виде белого твердого вещества. Этот материал направляли прямо на следующую стадию.(g) Preparation of 10-deacetyl-3'-N-desbenzoyl-3'-N- (tert-butyloxycarbonyl) -10-O - (phosphonooxymethyl) paclitaxel triethanolamine salt

A 500 ml Parr flask was charged with 10-deacetyl-2'-O-benzyloxycarbonyl-3'-N-desbenzoyl-3'-N- (tetra-butyloxycarbonyl) -10-O- (dibenzylphosphonoxymethyl) paclitaxel (264.9 mg, 0.215 mmol) and ethyl acetate (20 ml). Then the flask was blown with a stream of argon, and 10% Pd / C (318 mg) was added to it. The resulting mixture was placed on a Parr installation in a hydrogen atmosphere at a pressure of 55 psi. inch (3.86 kg / cm ² ). The reaction was monitored by HPLC (70:30 buffer CH ₃ CN / Q8, pH 6.0, 1.00 ml / min, Zorbax C-18 column, 25.0 cm, λ = 230 nm), until the starting material became visible (12.5 hours). The mixture was filtered through a pad of celite, which was washed with ethyl acetate and a small amount of dichloromethane. The resulting filtrate was concentrated, and the residue was dissolved in dichloromethane (5 ml). The addition of hexane caused a precipitate to form, from which 140.3 mg of free acid was isolated (purity, as determined by HPLC — 80%) as a white solid. This material was sent directly to the next stage.

Dichloromethane (10 ml) was added to the flask containing the above free acid (140 mg, 0.153 mmol). Then, the resulting solution was treated with a 0.100 M solution of triethanolamine in ethyl acetate (1.16 ml, 0.116 mmol), resulting in a cloudy solution. Approximately 2 ml of hexane was added and the mixture was kept at -20 ^° C. overnight. The resulting 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 to give 69.9 mg (42%) of the title triethanolamine salt as a gray powder, which had a purity of 95 - 96% as determined by HPLC analysis. (T _R = 2.05 min, Buffer 70:30
CH ₃ CN / Q8, pH 6.0, 1.00 ml / min, Zorbax C-18, 25.0 cm, λ = 230 nm).

¹ H NMR (d ₆ -acetone / D ₂ O, 300 MHz), δ: 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), 5.81 (s, 1H), 5.59 (d, J = 7.0 Hz, 1H), 5.12 (br, 2H), 4.93 ( d, J = 8.4 Hz, 2H), 4.56 (d, J = 4.9 Hz, 1H), 4.31 - 4.26 (m, 1H), 4.11 (s, 2H), 3.41 - 3.37 (m, 6H), 2.42 - 2.32 (m, 5H), 2.15 (br, 1H), 1.97 (s, 3H), 1.77 - 1, 64 (m, 2H), 1.58 (s, 3H), 1.13 (s, 9H), 1.15-1.07 (m, 6H).

¹³ C NMR (d ₆ -acetone, D ₂ O, 75.5 MHz), δ: 171.6, 166.9, 156.6, 141.8, 135.1, 134.2, 131.0, 130 7, 129.4, 129.3, 128.4, 128.1, 88.3, 85.4, 81.9, 79.7, 78.6, 78.1, 76.8, 76.0 , 74.8, 71.9, 71.2, 47.4, 44.0, 37.1, 36.3, 28.5, 27.0, 23.1, 22.0, 14.7, 10 ,4.

High-resolution mass spectrum: MNa ⁺ - 940.3142,
Calculated for C ₄₄ H ₅₆ NO ₁₈ PNa = 940.3133.

К раствору 7-O-триэтилсилилпаклитаксела (70,0 мг, 72,2 ммоль), бис (метилтиометил)эфира (90 мг, 72,2 ммоль), молекулярных сит (70 мг) и N-иодсукцинимида (160 мг, 72,2 ммоль) в тетрагидрофуране (2,0 мл) при комнатной температуре добавляли трифлат серебра (5,0 мг, 19,5 ммоль), и полученный раствор перемешивали 2 часа. Затем реакционную смесь разбавляли этилацетатом и фильтровали через слой целита. Фильтрат промывали насыщенным водным раствором бикарбоната натрия, а затем смесью (1:1, объем/объем) насыщенного водного раствора бикарбоната натрия и 5%-ного водного раствора тиосульфата натрия и наконец рассолом. Органику затем высушивали над сульфатом натрия и концентрировали в вакууме. Остаточное масло очищали путем флеш-хроматографии (3: 1, гексаны-этилацетат) и получили в результате 22,0 мг (29%) указанного в заголовке соединения в виде белого твердого вещества.Example 10. 2'-O-phosphonooxymethoxymethylpaclitaxel
(a) Preparation of 2'-O- (methylthiomethoxymethyl) -7-O-triethylsilylpaclitaxel

To a solution of 7-O-triethylsilylpaclitaxel (70.0 mg, 72.2 mmol), bis (methylthiomethyl) ether (90 mg, 72.2 mmol), molecular sieves (70 mg) and N-iodosuccinimide (160 mg, 72, 2 mmol) in tetrahydrofuran (2.0 ml), silver triflate (5.0 mg, 19.5 mmol) was added at room temperature, and the resulting solution was stirred for 2 hours. Then the reaction mixture was diluted with ethyl acetate and filtered through a pad of celite. The filtrate was washed with saturated aqueous sodium bicarbonate and then with a mixture (1: 1, v / v) of saturated aqueous sodium bicarbonate and 5% aqueous sodium thiosulfate and finally with brine. The organics were then dried over sodium sulfate and concentrated in vacuo. The residual oil was purified by flash chromatography (3: 1, hexanes-ethyl acetate) to give 22.0 mg (29%) of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.12 - 7.20 (15H, m), 7.04 (1H, d, J = 8.9 Hz), 6.41 (1H, s) 6.25 (1H, m), 5.81 (1H, dd, J = 8.9 and 2.4 Hz), 5.68 (1H, d, J = 7.0 Hz), 4.93 ( 1H, d, J = 8.0 Hz), 4.79 (2H, m), 4.71 (1H, d, J = 2.4 Hz), 4.45 (1H, dd, J = 10.5 and 6.6 Hz), 4.30 (1H, d, J = 8.3 Hz), 4.28 (1H, d, J = 11.7 Hz), 4.17 (1H, d, J = 8 , 3 Hz), 4.04 (1H, d, J = 11.7 Hz), 3.80 (1H, d, J = 6.9 Hz), 2.48 - 1.13 (25H, m, on singlets at 2.51, 2.13, 2.05, 2.01, 1.69, 1.19, 1.16), 0.98 - 0.85 (9H, m), 0.65 - 0 50 (6H, m).

К раствору полученного на стадии (a) продукта (15 мг, 0,0141 ммоль) и молекулярных сит (15 мг) в ТГФ (0,5 мл) при комнатной температуре добавляли дибензилфосфат (20,0 мг, 0,089 ммоль) и затем N-иодсункцинимид (4,2 мг, 0,0187 ммоль), и раствор перемешивали 1 час. В этот момент анализ реакционной смеси путем тонкослойной хроматографии показал присутствие только исходного материала. После этого добавляли тремя частями в течение 2 часов трифлат серебра (5,0 мг, 0,0189 ммоль), и реакционную смесь перемешивали еще 1 час. Затем реакционную смесь разбавляли этилацетатом, и полученный раствор фильтровали через слой целита. Фильтрат обрабатывали смесью (1: 1 объем/объем) насыщенного водного раствора бикарбоната натрия и 5%-ного водного раствора тиосульфата натрия. Органический экстракт затем промывали рассолом, высушивали над сульфатом натрия и концентрировали в вакууме. Остаточное масло очищали путем флеш-хроматографии (1:1, гексан:этилацетат) и получили 5,0 мг (33%) указанного в заголовке соединения.(b) Preparation of 2'-O- (dibenzylphosphonooxymethoxymethyl) -7-triethylsilylpaclitaxel

To a solution of the product obtained in stage (a) (15 mg, 0.0141 mmol) and molecular sieves (15 mg) in THF (0.5 ml), dibenzyl phosphate (20.0 mg, 0.089 mmol) was added at room temperature and then N iodosunkcinimide (4.2 mg, 0.0187 mmol) and the solution was stirred for 1 hour. At this point, analysis of the reaction mixture by thin layer chromatography showed the presence of only the starting material. After that, silver triflate (5.0 mg, 0.0189 mmol) was added in three parts over 2 hours, 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 celite pad. The filtrate was treated with a mixture (1: 1 v / v) of a saturated aqueous solution of sodium bicarbonate and a 5% aqueous solution of sodium thiosulfate. The organic extract was then washed with brine, dried over sodium sulfate and concentrated in vacuo. The residual oil was purified by flash chromatography (1: 1, hexane: ethyl acetate) to give 5.0 mg (33%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.08-7.16 (25H, m), 7.18 (1H, d, J = 8.8 Hz), 6.41 (1H, s) 6.21 (1H, m), 5.82 (1H, dd, J = 9.0 and 3.1 Hz), 5.66 (1H, d, J = 7 Hz), 5.01-4, 65 (10H, m), 4.56 (1H, dd, J = 14.7 and 5.6 Hz), 4.43 (1H, dd, J = 10; 4 and 6.7 Hz), 4.29 (1H, d, J = 8.3 Hz), 4.16 (1H, d, J = 8.3 Hz), 3.78 (1H, d, J = 7.0 Hz), 2.60 - 1 13 (22H, m, incl. Singles at 2.49, 2.15, 1.93, 1.66, 1.15, 1.13, 3H each), 0.95 - 0.84 (9H, m ), 0.63-0.45 (6H, m).

Полученный на стадии (b) продукт обрабатывали тетрабутиламмонийфторидом в соответствии с процедурой, описанной в примере 9 (f), для удаления 7-O-триэтилсилилзащитной группы. Полученное при этом соединение подвергали каталитической гидрогенизации в соответствии с процедурой, описанной в предшествующих примерах, и получили указанное в заголовке соединение.(c) Preparation of 2'-O-phosphonooxymethoxymethylpaclitaxel

The product obtained in step (b) was treated with tetrabutylammonium fluoride in accordance with the procedure described in Example 9 (f) to remove the 7-O-triethylsilyl protecting group. The compound thus obtained was subjected to catalytic hydrogenation in accordance with the procedure described in the previous examples, and the title compound was obtained.

К раствору паклитаксела (20,0 г, 0,0234 моль) и имидазола (3,59 г, 0,052 моль) в 150 мл ДМФ (диметилформамида) при 0^oC добавляли порциями по 2 мл в течение 20 мин триэтилсилилхлорид (6,0 мл, 0,053 моль). Затем реакционную смесь перемешивали при 0^oC в течение 1 часа. Далее смесь разбавляли этилацетатом и насыщенным водным раствором аммонийхлорида. Органический слой отделяли, промывали рассолом, высушивали над сульфатом натрия и концентрировали в вакууме, получив в результате желтое масло. Очистка неочищенного продукта путем флеш-хроматографии (гексаны:этилацетат, 1:3, а затем 1:1) дала 21,07 г (выход 98%) целевого указанного в заголовке соединения в виде неокрашенного белого твердого вещества.Example II 2'-O-phosphonooxymethoxymethylpaclitaxel (Alternative Method)
(a) Preparation of 2'-O-triethylsilylpaclitaxel

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 0 ^° C., 2 ml portions were added in 20 ml portions over 20 minutes (6.0 ml, 0.053 mol). Then the reaction mixture was stirred at 0 ^o C for 1 hour. The mixture was then diluted with ethyl acetate and saturated aqueous ammonium chloride. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated in vacuo, resulting in a yellow oil. Purification of the crude product by flash chromatography (hexanes: ethyl acetate, 1: 3 and then 1: 1) gave 21.07 g (98% yield) of the desired title compound as an unpainted white solid.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.15 (2H, m), 7.70 (2H, m), 7.65 - 7.30 (11H, m), 7.15 (1H, d, J = 8.9 Hz), 6.30 (1H, s), 6.25 (1H, m), 6.70 - 6.10 (2H, m), 4.94 (1H, d, J = 7.9 Hz), 4.67 (1H, d, J = 2.0 Hz), 4.40 (1H, m), 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).

К раствору 2'-O-триэтилсилилпаклитаксела (22,3 г, 24,1 ммоль) в ТГФ (250 мл), охлажденном до -50^oC, добавляли по каплям в течение 10 минут бутиллитий (1,6 М в гексанах, 12,9 мл, 8,06 ммоль). Полученный раствор перемешивали 20 мин, поддерживая при этом температуру в пределах между -50^oC и -35^oC. Затем реакционную смесь охлаждали до -50^oC и добавляли в нее по каплям в течение 5 минут бензилхлорформиат (5,0 мл, 29,8 ммоль). Реакционную смесь выдерживали при -40^oC в течение 30 минут, после чего приводили в равновесие до 0^oC в течение приблизительно 30 минут. Затем смесь разбавляли этилацетатом и насыщенным водным раствором аммоний хлорида, и полученный органический слой промывали рассолом, высушивали над сульфатом натрия и концентрировали в вакууме. ¹H ЯМР-анализ неочищенной смеси показал наличие требуемого 2'-O-триэтилсилил-7-O-бензилоксикарбонилпаклитаксела, а также 2'-O-триэтилсилил-7-эпигидроксипаклитаксела (отношение 3: 1 соответственно). Эту смесь продуктов использовали на следующей стадии без дальнейшей очистки и затем разделения изомеров. Аналитическую пробу основного продукта 2'-O-триэтилсилил-7-O-бензилоксикарбонилпаклитаксела очищали путем флеш-хроматографии.(b) Preparation of 2'-O-triethylsilyl-7-O-benzyloxycarbonylpaclitaxel

To a solution of 2'-O-triethylsilylpaclitaxel (22.3 g, 24.1 mmol) in THF (250 ml), cooled to -50 ^° C, butyllithium (1.6 M in hexanes, 12) was added dropwise over 10 minutes. 9 ml, 8.06 mmol). The resulting solution was stirred for 20 minutes while maintaining the temperature between -50 ^o C and -35 ^o C. Then the reaction mixture was cooled to -50 ^o C and benzyl chloroformate (5.0 ml, 29) was added dropwise thereto over 5 minutes. , 8 mmol). The reaction mixture was kept at -40 ^° C for 30 minutes, after which it was equilibrated to 0 ^° C for approximately 30 minutes. The mixture was then diluted with ethyl acetate and a saturated aqueous solution of ammonium chloride, and the resulting organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. ¹ H NMR analysis of the crude mixture revealed the desired 2'-O-triethylsilyl-7-O-benzyloxycarbonylpaclitaxel as well as 2'-O-triethylsilyl-7-epihydroxypaclitaxel (3: 1 ratio, respectively). This product mixture was used in the next step without further purification and then isomer separation. An analytical sample of the main product of 2'-O-triethylsilyl-7-O-benzyloxycarbonyl-paclitaxel was purified by flash chromatography.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.12 (2H, m), 7.72 (1H, m), 7.65 - 7.27 (1H, d, J = 8.8 Hz) , 6.41 (1H, m), 6.20 (1H, m), 5.72 - 5.65 (2H, m), 5.52 (1H, m), 5.24 (1H, d, J = 12.3 Hz), 5.16 (1H, d, J = 12.3 Hz), 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), 3.94 (1H, d, J = 6.8 Hz), 2.70 - 1.12 (22H, incl. Singlets at 2.54, 2.14, 2.01, 1.80, 1.20, 1.15, 3H each), 0.81 - 0.73 (9H, m) , 0.55-0.31 (6H, m).

К раствору полученного на стадии (b) продукта (24,0 г, 22,6 ммоль) в ацетонитриле (250 мл), охлажденном до 0^oC, добавляли хлористоводородную кислоту (6 н., 1,0 мл, 6,0 ммоль). Через 10 минут анализ путем тонкослойной хроматографии (гексаны: этилацетат, 1: 1) показал, что реакция завершена. Реакционную смесь разбавляли насыщенным водным раствором бикарбоната натрия, а затем этилацетатом и органический слой отделяли, промывали рассолом, высушивали сульфатом натрия и концентрировали в вакууме. Остаточное масло очищали путем флеш-хроматографии (гексаны:этилацетат, 1:3, а затем 1:1) и получили в результате 11,4 г (48% за две стадии) указанного в заголовке соединения и 4,8 г (20%) 7-эпигидроксипаклитаксела.(c) Preparation of 7-O-benzyloxycarbonylpaclitaxel

To a solution of the product obtained in stage (b) (24.0 g, 22.6 mmol) in acetonitrile (250 ml), cooled to 0 ^° C., hydrochloric acid (6 N, 1.0 ml, 6.0 mmol) was added ) After 10 minutes, analysis by thin layer chromatography (hexanes: ethyl acetate, 1: 1) showed that the reaction was completed. The reaction mixture was diluted with saturated aqueous sodium bicarbonate, and then ethyl acetate and the organic layer were separated, washed with brine, dried with sodium sulfate and concentrated in vacuo. The residual oil was purified by flash chromatography (hexanes: ethyl acetate, 1: 3 and then 1: 1) to give 11.4 g (48% in two steps) of the title compound and 4.8 g (20%) 7-epihydroxypaclitaxel.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 8.09 (2H, m), 7.71 (2H, m), 7.65 - 7.27 (16H, m), 7.10 (1H, d, J = 8.9 Hz), 6.39 (1H, s), 6.16 (1H, m), 5.81 (1H, d, J = 8.9 and 2.4 Hz), 5, 65 (1H, d, J = 6.9 Hz), 5.49 (1H, dd, J = 10.6 and 7.2 Hz), 5.20 (1H, d, J = 11.9 Hz), 5.12 (1H, d, J = 11.9 Hz), 4.91 (1H, d, J = 8.4 Hz), 4.78 (1H, m), 4.30 (1H, d, J = 8.4 Hz), 4.15 (1H, d, J = 8.4 Hz), 3.91 (1H, d, J = 6.8 Hz), 3.39 (1H, d, J = 4 9 Hz), 2.65 - 1.10 (22H, incl. Singlets at 2.39, 2.18, 1.81, 1.75, 1.21, 1.15, 3H each).

К раствору 7-O-бензилоксикарбонилпаклитаксела (5,53 г, 5,71 ммоль), простого 1,1'-дитиометилдиметилового эфира (7,8 г, 57,1 ммоль), N-иодсукцинимида (6,35 г, 28,3 ммоль) и высушенных в печи порошкообразных молекулярных сит (5,0 г), в ТГФ (110 мл) при комнатной температуре добавляли трифлат серебра (300 мг, 1,17 ммоль). Анализ путем тонкослойной хроматографии (гексаны:этилацетат, 1:1) реакционной смеси через 20 минут показал превращение примерно 40% исходного материала в высшие фракции. Затем добавляли трифлат серебра (150 мг, 0,585 ммоль), и контролировали реакцию путем тонкослойной хроматографии, которая показала, что спустя 30 минут реакция была завершена приблизительно на 65%. Смесь разбавляли этилацетатом (100 мл), фильтровали через слой целита, и фильтрат выливали в делительную воронку, содержавшую 20 мл насыщенного водного раствора бикарбоната натрия и 50 мл 5%-ного водного раствора тиосульфата натрия. Органический слой отделяли, промывали рассолом, высушивали над сульфатом натрия и концентрировали в вакууме. Остаточное масло очищали путем флеш-хроматографии (гексаны:этилацетат, градиентное элюирование, от 4:1 до 3:2) и получили 3,0 г (выход 54%) указанного в заголовке соединения в виде светло-желтого твердого вещества.(d) Preparation of 2'-O- (methylthiomethoxymethyl) -7-O-benzyloxycarbonylpaclitaxel

To a solution of 7-O-benzyloxycarbonylpaclitaxel (5.53 g, 5.71 mmol), 1,1'-dithiomethyldimethyl ether (7.8 g, 57.1 mmol), N-iodosuccinimide (6.35 g, 28, 3 mmol) and oven-dried powder molecular sieves (5.0 g), silver triflate (300 mg, 1.17 mmol) was added at room temperature in THF (110 ml). Analysis by thin layer chromatography (hexanes: ethyl acetate, 1: 1) of the reaction mixture after 20 minutes showed the conversion of approximately 40% of the starting material to higher fractions. Then silver triflate (150 mg, 0.585 mmol) was added, and the reaction was monitored by thin layer chromatography, which showed that after 30 minutes the reaction was completed by about 65%. The mixture was diluted with ethyl acetate (100 ml), filtered through a pad of celite, and the filtrate was poured into a separatory funnel containing 20 ml of a saturated aqueous solution of sodium bicarbonate and 50 ml of a 5% aqueous solution of sodium thiosulfate. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated in vacuo. The residual oil was purified by flash chromatography (hexanes: ethyl acetate, gradient elution, 4: 1 to 3: 2) to give 3.0 g (54% yield) of the title compound as a pale yellow solid.

¹ H NMR (CDCl ₃ , 300 MHz), δ: 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), 6.26 (1H, m), 5.77 (1H, dd, J = 8.8 and 2.5 Hz), 5 , 71 (1H, d, J = 6.9 Hz), 5.51 (1H, dd, J = 10.6 and 7.1 Hz), 5.21 (1H, d, J = 11.9 Hz) 5.14 (1H, d, J = 11.9 Hz), 4.92 (2H, m), 4.68 (1H, d, J = 2.5 Hz), 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, including singlets at 2.50, 2.15, 2.05, 1, 74, 1.72, 1.20, 1.15, 3H each).

К раствору 2'-O-(метилтиометоксиметил)-7-O-бензилоксикарбонилпаклитаксела (1,06 г, 1,07 ммоль) и высушенных в печи порошкообразных молекулярных сит (1,0 г), в ТГФ (20 мл) при комнатной температуре дибензилфосфат (1,49 г, 5,30 ммоль), а затем сразу же N-иодсукцинимид (2,65 г, 1,18 ммоль). Анализ путем тонкослойной хроматографии (гексаны:этилацетат, 1:1) реакционной смеси через 2,5 часа показал, что реакция завершена приблизительно на 60%. После этого добавляли N-иодсукцинимид (175 мг, 0,78 ммоль), и реакционную смесь перемешивали еще 30 минут, после чего анализ путем тонкослойной хроматографии показал, что реакция завершена. Далее реакционную смесь разбавляли этилацетатом (50 мл) и фильтровали через слой целита. Фильтрат выливали в делительную воронку, содержавшую 100 мл насыщенного раствора бикарбоната натрия и 20 мл 5%-ного водного раствора тиосульфата натрия. Органический слой отделяли, промывали рассолом, высушивали над сульфатом натрия, и концентрировали в вакууме. Остаточное масло очищали путем флеш-хроматографии (гексаны:этилацетат, градиентное элюирование, от 3:1 до 1:1) и получили 750 мг (62%-ный выход целевого указанного в заголовке соединения в виде белого твердого вещества.(e) Preparation of 2'-O-dibenzylphosphonoxymethoxymethyl) -7-O-benzyloxycarbonylpaclitaxel

To a solution of 2'-O- (methylthiomethoxymethyl) -7-O-benzyloxycarbonylpaclitaxel (1.06 g, 1.07 mmol) and oven-dried powder molecular sieves (1.0 g), in THF (20 ml) at room temperature dibenzyl phosphate (1.49 g, 5.30 mmol) and then immediately N-iodosuccinimide (2.65 g, 1.18 mmol). Analysis by thin layer chromatography (hexanes: ethyl acetate, 1: 1) of the reaction mixture after 2.5 hours showed that the reaction was completed by approximately 60%. After that, N-iodosuccinimide (175 mg, 0.78 mmol) was added, and the reaction mixture was stirred for another 30 minutes, after which analysis by thin-layer chromatography showed that the reaction was completed. Next, the reaction mixture was diluted with ethyl acetate (50 ml) and filtered through a pad of celite. The filtrate was poured into a separatory funnel containing 100 ml of a saturated solution of sodium bicarbonate and 20 ml of a 5% aqueous solution of sodium thiosulfate. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residual oil was purified by flash chromatography (hexanes: ethyl acetate, gradient elution, 3: 1 to 1: 1) to give 750 mg (62% yield of the title compound as a white solid.

¹ H NMR (CDCl ₃ , 360 MHz), δ: 8.10 (2H, m), 7.79 (2H, m), 7.65 - 7.24 (26H, m), 7.10 (1H, m), 6.41 (1H, s), 6.20 (1H, m), 5.79 (1H, dd, J = 8.8 and 3.6 Hz), 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 - 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, including singlets at 2.48, 2.19, 1.95, 1, 80, 1.20, 1.10, 3H each).

(f) preparation of the triethanolamine salt of 2'-O-phosphonooxymethoxymethylpaclitaxel
To a solution of 2'-O- (dibenzylphosphonooxymethoxymethyl) -7-O-benzyloxycarbonylpaclitaxel (500 mg, 0.382 mmol) in ethyl acetate (40 ml) in a Parr flask was added palladium (10%) on carbon. The flask was mounted on a Parr apparatus, and the reaction mixture was exposed to hydrogen at 50 psi. inch (3.5 kg / cm ² ). The reaction mixture was shaken for 6.5 hours and then filtered through a funnel with a porous glass plate. Triethanolamine (0.1 N in ethyl acetate, 4.0 ml) was added to this filtrate, and the resulting solution was concentrated in vacuo. The crude solid was suspended in about 5.0 ml of ethyl acetate, and the solvent was decanted. This process was repeated three times, whereby the title triethanolamine salt (300 mg) was obtained with a purity of 87% as determined by HPLC analysis. Further purification of this compound by chromatography on a C18 column (water: acetonitrile, 3: 1) gave the title target compound (120 mg, 34%) with a purity of 95% as determined by HPLC.

¹ H NMR (CD COCD ₃ , D ₂ O, 300 MHz), δ: 9.05 (1H, d, J = 8.7 Hz), 8.15 - 7.12 (21H, m), 6.40 (1H, m), 6.05 (1H, m), 5.69 - 5.55 (2H, m), 5.01 - 4.85 (6H, m), 4.35 (1H, m), 4.14 (2H, m), 3.96 - 3.85 (6H, m), 3.25 (1H, d, J = 7.1 Hz).

Claims (54)

1. Phosphonoxymethyl esters of taxon derivatives of the formula

where R ¹ is hydroxy-OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or OC (O) OR ^x ;
R ^{2 '} is hydrogen and R ² is hydroxy, -OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ or -OC (O) OR ^x or R ^2' is fluorine,
R ² is hydrogen;
R ³ is hydrogen, hydroxy, acetoxy;
R ⁴ , R ⁵ - independently (C ₁ - C ₆ ) alkyl or Z - R ^{6 '} ;
Z is a direct link;
R ^{6 '} is aryl or heteroaryl,
R ^x - (C ₁ - C ₆ ) alkyl;
p is 0 or 1;
R ⁶ and R ⁷ together form an oxo group, provided that at least one of R ¹ , R ² is OCH ₂ (OCH ₂ ) _m OP (O) (OH) ₂ ;
m = 0 or 1,
or their pharmaceutically acceptable salts. 2. The compound according to claim 1, where R ^{2 '} is hydrogen and R ² is OCH ₂ OP (O) (OH) ₂ , or a pharmaceutically acceptable salt thereof. 3. The compound of claim 2, wherein R ¹ is hydroxy or OC (O) OR ^x and R ^x is (C ₁ - C ₆ ) alkyl. 4. The compound according to claim 3, where R ³ is hydrogen, hydroxy or acetoxy. 5. The compound according to claim 3, where R ⁴ (O) _p is phenyl or tertbutoxy. 6. The compound according to claim 3, where R ⁵ is phenyl, 2-furyl or 2-thienyl.  7. The compound according to claim 1, which is 2'-0- (ethoxycarbonyl) -7-0- (phosphonoxymethyl) paclitaxel or a pharmaceutically acceptable salt thereof.  8. The compound according to claim 7, which is its sodium salt.  9. The compound according to claim 7, which is its triethanolamine salt.  10. The compound according to claim 7, which is its triethylamine salt.  11. The compound according to claim 7, which is its arginine salt.  12. The compound according to claim 7, which is its lysine salt.  13. The compound according to claim 7, which is its ethanolamine salt.  14. The compound according to claim 7, which is its N-methylglucamine salt.  15. The compound according to claim 7, which is 7-0- (phosphonooxymethyl) paclitaxel or a pharmaceutically acceptable salt thereof.  16. The compound according to clause 15, which is its sodium salt.  17. The compound according to claim 1, which is 3'-N-debenzoyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -2'-0-ethyloxycarbonyl-7-0-phosphonoxymethylpaclitaxel or a pharmaceutically acceptable salt thereof.  18. The compound according to 17, which is its triethanolamine salt.  19. The compound according to claim 1, which is 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -2'-0-ethyloxycarbonyl- 7-0-phosphonooxymethylpaclitaxel or a pharmaceutically acceptable salt thereof.  20. The compound according to claim 19, which is its triethanolamine salt. 21. The compound according to claim 1, where R ¹ -OCH ₂ OP (O) (OH) ₂ , or its pharmaceutically acceptable salt. 22. The compound according to item 21, where R ^{2 '} is hydrogen, R ² is hydrogen, hydroxy or —OC (O) OR ^x and R ^x is (C ₁ - C ₆ ) alkyl. 23. The compound according to item 22, where R ³ is hydrogen, hydroxy or acetoxy. 24. The compound of claim 22, wherein R ⁴ (O) _p is phenyl or tert-butoxy. 25. The compound according to item 22, where R ⁵ is phenyl.  26. The compound according to claim 1, which is 2-0- (phosphonooxymethyl) paclitaxel or a pharmaceutically acceptable salt thereof. 27. The compound according to claim 1, where R ¹ and R ² are both OCH ₂ OP (O) (OH) ₂ , or a pharmaceutically acceptable salt thereof.  28. The compound according to claim 1, which is 2 ', 7-0-bis (phosphonooxymethyl) paclitaxel or a pharmaceutically acceptable salt thereof.  29. The compound of claim 28, which is its sodium salt. 30. The compound according to claim 1, where R ¹ -OCH ₂ OCH ₂ OP (O) (OH) ₂ , or its pharmaceutically acceptable salt.  31. The compound according to claim 1, which is 2'-0-phosphonooxymethoxymethylpaclitaxel or a pharmaceutically acceptable salt thereof.  32. The compound according to p, which is its triethanolamine salt.  33. The compound according to claim 1, which is 10-deacetyl-3'-N-desbenzoyl-3'-N- (tert-butyloxycarbonyl) -10-0- (phosphonooxymethyl) paclitaxel or a pharmaceutically acceptable salt thereof.  34. The compound of claim 33, which is its triethanolamine salt. 35. 7-0-methylthiomethylbaccatin III of the formula

or its C ₁₃ metal alkoxide. 36. Phosphonoxymethyl esters of taxane derivatives of the formula

where R ^1b is hydroxy, OCH ₂ SCH ₃ or OC (O) OR ^x ;
R ^{2 ′} is hydrogen or R ^2b is hydroxy, hydroxy, protected with a phosphono group —OCH ₂ S CH ₃ or R ^{2 ′} is fluoro, and R ^2b is hydrogen;
R ^3b is hydroxy protected by an acetoxy trialkylsilyl group;
R ⁴ , R ⁵ - independently (C ₁ - C ₆ ) alkyl or Z - R ^{6 '} ;
R ^{6 '} is aryl or heteroaryl;
R ^x - (C ₁ - C ₆ ) alkyl,
p is 0 or 1;
R ^6b and R ^7b together form an oxo group, provided that at least one of R ^1b , R ^2b is OCH ₂ SCH ₃ .  37. The compound according to clause 36, which is a 7-0-methylthiomethylpaclitaxel.  38. The compound according to clause 36, which is 2'-0- (benzoyloxycarbonyl) -7-0-methylthiomethylpaclitaxel.  39. The compound according to clause 36, which is 2'-0- (ethoxycarbonyl) - 7-0-methylthiomethylpaclitaxel.  40. The compound according to clause 36, which is 2'-0- (methylthiomethyl) -7-0- (triethylsilyl) -paclitaxel.  41. The compound of claim 36, which is 2-0- (methylthiomethyl) paclitaxel.  42. The compound according to clause 36, which is 2 ', 7-0-bis (methylthiomethyl) paclitaxel.  43. The compound according to clause 36, which is 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -7-0-methylthiomethylpaclitaxel.  44. The compound according to clause 36, which is 3'-N-debenzoyl-3'-desphenyl-3-N- (tert-butyloxycarbonyl) -3 '- (2-furyl) -2'-0-ethyloxycarbonyl-7 -0-methylthiomethylpaclitaxel.  45. The compound according to clause 36, which is 3'-N-debenzoyl-3'-desphenyl-3'-N) (tert-butyloxycarbonyl) -3 '- (2-thienyl) -7-0-methylthiomethylpaclitaxel.  46. The compound according to clause 36, which is 3'-N-debenzoyl-3'-desphenyl-3'-N- (tert-butyloxycarbonyl) -3 '- (2-thienyl) -2'-0-ethyloxycarbonyl- 7-0-methylthiomethylpaclitaxel. 47. The compound of the formula

where R ^{2 '} , R ^2b , R ^3b , R ⁴ , R ⁵ , R ^6b , R ^7b and p are defined above.  48. The compound of claim 47, which is 2'-0- (methylthiomethoxymethyl) -7-0-triethylsilylpaclitaxel.  49. The compound of claim 47, which is 2'-0- (methylthiomethoxymethyl) -7-0-benzyloxycarbonylpaclitaxel. 50. Phosphonoxymethyl esters of taxane derivatives of the formula

where R ^1k is hydroxy, -OCH ₂ OP (O) (OCH ₂ R ^y ) ₂ or -OC (O) OR ^x ;
R ^{2 ′} is hydrogen, R ^2c is hydroxy, hydroxy is protected by a trialkylsilyl group, OCH ₂ OP (O) (OCH ₂ R ^y ) ₂ or R ^{2 ′} is fluoro,
R ^2c is hydrogen; R ^3c is acetoxy; R ^x is a phosphonoprotective group;
R ^6c and R ^7c together form an oxo group, provided that at least one of R ^1c , R ^2c is —OCH ₂ OP (O) (OCH ₂ R ^y ) ₂ ;
R ⁴ , R ⁵ - independently (C ₁ - C ₆ ) alkyl or Z - R ⁶ ;
z is a direct connection;
R ^{6 '} is aryl or heteroaryl;
R ^x - (C ₁ - C ₆ ) alkyl,
p is 0 or 1;
R ^y is a phosphonoprotective group. 51. The compound of the formula

where R ^{2 '} , R ^2c , R ^3c , R ⁴ , R ⁵ , R ^6c , R ^7c , R ^y and p are as defined above.  52. An antitumor pharmaceutical composition comprising an active principle represented by a taxane derivative and pharmaceutically acceptable carriers, wherein the taxon derivative contains an effective antitumor amount of a compound according to claim 1.  53. A method of inhibiting tumor growth in a mammal, comprising administering an antitumor agent to the mammal, characterized in that an effective amount of a compound according to claim 1 is administered as an antitumor agent.  54. The method according to p. 50, characterized in that it includes the oral administration of an antitumor agent to the body. Priority on points:
12.24.92 - according to claims 1-17, 22-31, 34, 37-39, 47;
08/17/93 - the rest of the claims.

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