RU2195458C2 - Cryptoficine compound, pharmaceutical composition, method of inhibition of cell proliferation and method of pathological state relief - Google Patents

Abstract

FIELD: organic chemistry, medicine, oncology, pharmacy. SUBSTANCE: invention relates to a novel cryptoficine compound of the formula (I)

where Ar means methyl or phenyl or any simple unsubstituted or substituted aromatic or heterocyclic group; R₁ means halogen atom; R₂ means OH; or R₁ and R₂ can be joined to form epoxy-ring, aziridine ring; R₁ and R₂ can be joined in common to form a double bond between C₁₈ and C₁₉; R₃ means lower alkyl group; R₄ and R₅ mean H; or R₄ and R₅ can be joined to form a double bond between C₁₃ and C₁₄; R₆ means benzyl, hydroxybenzyl (oxybenzyl), alkoxybenzyl, halogenoxybenzyl, dihalogenoxybenzyl, halogenalkoxy-benzyl or dihalogenalkoxybenzyl group; R₇, R₈, R₉, R₁₀ each means independently H or lower alkyl group; X and Y each means independently O, NH. Compounds can be used for inhibition of proliferation of cells in mammal and for treatment of tumors. Invention describes also pharmaceutical composition based on compounds and methods of using. EFFECT: improved method of treatment, valuable medicinal properties of compounds. 56 cl, 10 tbl, 14 dwg, 22 ex

Description

 This invention was implemented in part with the support of the US Government with grants No. CA 12623 and CA 53001 from The National Cancer Institute, Department of Health and Human Services. Thus, the US government may have certain rights to this invention.

 Neoplastic diseases characterized by cell proliferation do not undergo normal control of cell growth, most of them cause death in people. A clinical chemotherapy trial has shown that new and more effective drugs are desirable for treating these diseases. Such an experiment also demonstrated that drugs that destroy the cytoskeletal microtubule system can be effective in inhibiting tumor cell proliferation.

 The microtubule system of eukaryotic cells is the main component of the cytoskeleton and is in a dynamic state - an ordered structure and a disordered structure; Thus, tubulin heterodimers polymerize with the formation of microtubules, and microtubules are depolymerized to their constituent components. Microtubules play a key role in the regulation of cellular architecture, metabolism and division. The dynamic state of microtubules is critical to their normal functions. As for cell division, tubulin polymerizes into microtubules, which form a mitotic spindle. The microtubules are then depolymerized when the mitotic spindle used is full. Thus, agents that disrupt the polymerization or depolymerization of microtubules, and thereby inhibit mitosis, include some of the most effective chemotherapeutic agents for clinical use.

 Such antimitotic agents or poisons can be classified into three groups based on their molecular mechanism of action. The first group consists of agents including colchicine and colcemid, which inhibit the formation of microtubules due to the destruction of tubulin. The second group consists of agents including vinblastine and vincristine, which induce the formation of paracrystalline tubulin aggregates. Vinblastine and vincristine are well known anti-cancer drugs. Their effect of destroying the microtubules of the mitotic spindle mainly inhibits hyperproliferative cells. The third group consists of agents including taxol, which accelerates the polymerization of tubulin and thus stabilizes the structure of microtubules.

 However, most of them, which have activity as an antitumor agent, do not guarantee efficacy against a tumor cell and, of course, not against a tumor cell that exhibits a phenotype of drug resistance. Wink alkaloids, such as vinblastine and vincristine, are effective against tumor cells and tumors, however, they lack activity against some tumors and cells with drug resistance. One of the foundations of a tumor cell exhibiting drug resistance (DR) or multiple drug resistance (MDR) is continuous over-expression of P-glycoprotein. Compounds that are poor substrates for the transport of P-glycoprotein should be suitable for the destruction of such DR or MDR phenotypes.

 Thus, the manifestation of the DR or MDR phenotype by many tumor cells and the clinically proven form of action of anti-microtubule agents against tumor cells makes it necessary to develop anti-microtubule cytotoxic agents in relation to tumor cells with no drug resistance, as well as cytotoxic in relation to tumor cells with a drug resistance phenotype. Agents that are promising in this regard include a class of compounds known as cryptophycins.

 Regarding methods for producing cryptophycins, a method for the complete synthesis of cryptophycins is not currently known. Cryptophycin compounds are currently obtained by isolation from blue-green algae or by semi-synthetic process variants of such naturally prepared compounds. The lack of a complete synthetic method inevitably makes it difficult to obtain stereospecific cryptophycins, a method that can lead to maximum activity and increase the stability of the compound. For example, a study showed that cryptophycins with an inactive macrocyclic ring are more active. Thus, a complete synthetic method that can produce cryptophycins with a macrocyclic ring, which is more stable than in naturally obtained cryptophycins, will be desirable. The present invention solves these problems.

где Аr представляет метил или фенил, или любую простую незамещенную, или замещенную ароматическую, или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между C₁₈ и C₁₉;
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между С₁₃ и C₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇, R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
Х и Y каждый независимо представляют О, NH или алкиламино.The present invention provides new cryptophycin compounds having the following structure:

where Ar represents methyl or phenyl, or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

 The present invention further provides general synthetic methods for producing cryptophycins. The present invention also provides the use of cryptophycins in pharmaceutical preparations for inhibiting the proliferation of mammalian cells and for treating a tumor.

Figure 1 represents the general structure of selected cryptophycin compounds of the present invention and a number system for hydroxy acid elements A and D and amino acid elements B and C in selected embodiments;
figures 2A and 2B graphically depict the effects of cryptophycin compounds and vinblastine on Jurkat cell proliferation and cell cycle progression. Jurkat cells were incubated with the indicated concentrations of cryptophycin compounds (A) or vinblastine (B) for 24 hours. For each sample, the number of viable cells (■) and the mitotic index (□) were determined, as described in the experimental part. Values represent the value +/- standard deviation (sd) for three samples in one of three such experiments;
figure 3 graphically depicts the reversibility of the effects of vinblastine, cryptophycin and taxol on cell growth. SKOV3 cells were treated with 0.1 nM vinblastine (□), 0.1 nM cryptophycin (■) or 1 nM taxol (x) at time = 0. These concentrations inhibited cell growth by 50% for each compound. After 24 hours, the cells were washed and incubated in a drug-free medium for the indicated time. Cell density was determined by sulphorodamine B (SRB) staining as described in the experimental part and expressed as +/- sd absorbance at 560 nm for ternary samples in one of three experiments;
figure 4 depicts isobolograms for the combination effects of vinblastine and cryptophycin on cell proliferation. SKOV3 cells were treated with vinblastine (0-600 pM) and / or cryptophycin (1-100 pM) for 48 hours. Then, the number of cells was determined using SRB staining, as described in the experimental part, and the IC _50s value (■) and the additivity line (---) for combinations of vinblastine and cryptophycin compounds were determined. Values represent values for two experiments, each containing triple samples;
figure 5 is a first cryptophycin synthesis scheme in accordance with the present invention;
figure 6 is a diagram for obtaining oxyacid element A;
Figure 7 is a diagram for producing a cryptophycin subunit comprising an hydroxy acid element A and amino acid B;
figure 8 is a diagram for producing a cryptophycin subunit comprising amino acid element C and hydroxy acid D;
figure 9 represents the first scheme for the synthesis of selected cryptophycins in accordance with the present invention;
figure 10 represents a second scheme for the synthesis of selected cryptophycins in accordance with the present invention;
figure 11 is a diagram for the synthesis of a subunit of cryptophycin, including hydroxy acid D;
figure 12 represents a third scheme for the synthesis of selected cryptophycins in accordance with the present invention;
figure 13 is a fourth diagram for the synthesis of selected cryptophycins in accordance with the present invention; and
figure 14 represents a fifth scheme for the synthesis of selected cryptophycins in accordance with the present invention.

где Аr представляет метил, или фенил, или любую простую незамещенную или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между C₁₈ и C₁₉.The present invention provides new cryptophycin compounds having the following structure:

where Ar represents methyl, or phenyl, or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ .

R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

где R₁ представляет Н, ОН, галоид, О кетогруппы, NH₂, SH, низшую алкокси группу или низшую алкильную группу;
R₂ представляет Н, ОН, О кетогруппы, NH₂, SH, низшую алкокси группу или низшую алкильную группу; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца азиридинового кольца, эписульфидного кольца, или двойной связи между С₁₀ и С₁₁; или
R₁ и R₂ могут объединяться вместе с образованием тетрагидрофуранового кольца;
R₃ представляет Н или низшую алкильную группу;
R₄ представляет ОН, низшую алканоильную группу или низшую α-оксиалканоилокси группу;
R₅ представляют Н или ОН группу;
R₆ представляет Н; или
R₅ и R₆ могут объединяться вместе с образованием двойной связи между С₅ и С₆;
R₇ представляет бензильную, оксибензильную, метоксиксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидметоксибензильную или дигалоидметоксибензильную группу;
R₈ представляет ОН, низшую β-аминокислоту, С₁ связывается с N β-аминокислоты, или этерифицированной низшей β-аминокислоты, где С₁ связывается с N этерифицированной низшей β-аминокислотной группы;
R₄ и R₈ могут объединяться вместе с образованием дидепсипептидной группы, состоящей из низшей β-аминокислоты, связанной с низшей α-оксиалкановой кислотой; и
R₅ и R₈ могут объединяться вместе с образованием дидепсипептидной группы, состоящей из низшей

-аминокислоты, связанной с низшей α-оксиалкановой кислотой; и
со следующими условиями:
R₁ представляет Н, низшую алкильную группу или низшую алкокси группу только, если R₂ представляет ОН, О кетогруппы, NH₂, SH;
R₂ представляет Н, низшую алкильную группу или низшую алкокси группу только, если R₁ представляет ОН, О кетогруппы, NH₂, SH;
если R₁ представляет ОН, R₂ представляет ОН, R₃ представляет метил, R₅ и R₆ могут объединяться вместе с образованием двойной связи между C₅ и C₆, R₄ и R₈ объединяются вместе с образованием дидепсипептидной группы со структурой X:

где O₁ указанного X соответствует R₄, N₈ указанного Х соответствует R₈, R₉ представляет метил и R₁₀ представляет изобутил, R₇ не является 3-хлор-4-метоксибензилом;
если R₁ и R₂ объединяются вместе с образованием эпоксидного кольца, R₉ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, R₉ представляет метил и R₁₀ представляет изобутил, R₇ не является 3-хлор-4-метоксибензилом;
если R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, R₉ представляет метил и R₁₀ представляет изобутил, R₇ не является 3-хлор-4-метоксибензилом; и
если R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₉ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₄ связывается с концевой карбоксильной группой лейциновой кислоты и R₈ присоединяется к азоту концевой группы либо 3-амино-2-метилпропионовой кислоты, либо метилового эфира 3-амино-2-метилпропионовой кислоты, R₇ не является 3-хлор-4-метоксибензилом.In one aspect of the present invention, new cryptophycin compounds are provided having the following structure:

where R ₁ represents H, OH, halogen, O keto groups, NH ₂ , SH, a lower alkoxy group or a lower alkyl group;
R ₂ represents H, OH, O ketogroups, NH ₂ , SH, a lower alkoxy group or a lower alkyl group; or
R ₁ and R ₂ may combine to form an epoxy ring of the aziridine ring, an episulfide ring, or a double bond between C ₁₀ and C ₁₁ ; or
R ₁ and R ₂ may combine to form a tetrahydrofuran ring;
R ₃ represents H or a lower alkyl group;
R ₄ represents OH, a lower alkanoyl group or a lower α-hydroxyalkanoyloxy group;
R ₅ represent H or OH group;
R ₆ represents H; or
R ₅ and R ₆ may combine to form a double bond between C ₅ and C ₆ ;
R ₇ represents a benzyl, oxybenzyl, methoxybenzyl, halooxybenzyl, dihalooxybenzyl, halogenomethoxybenzyl or dihalomethoxybenzyl group;
R ₈ represents OH, a lower β-amino acid, C ₁ binds to an N β-amino acid, or an esterified lower β-amino acid, where C ₁ binds to an N esterified lower β-amino acid group;
R ₄ and R ₈ can combine to form a didepsipeptide group consisting of a lower β-amino acid linked to a lower α-hydroxyalkanoic acid; and
R ₅ and R ₈ may combine to form a didepsipeptide group consisting of a lower

-amino acids bound to lower α-hydroxyalkanoic acid; and
with the following conditions:
R ₁ represents H, a lower alkyl group or a lower alkoxy group only if R ₂ represents OH, O keto groups, NH ₂ , SH;
R ₂ represents H, a lower alkyl group or a lower alkoxy group only if R ₁ represents OH, O keto groups, NH ₂ , SH;
if R ₁ represents OH, R ₂ represents OH, R ₃ represents methyl, R ₅ and R ₆ can combine together to form a double bond between C ₅ and C ₆ , R ₄ and R ₈ combine to form a didepsipeptide group with structure X:

where O _{1 of the} specified X corresponds to R ₄ , N _{8 of the} specified X corresponds to R ₈ , R ₉ represents methyl and R ₁₀ represents isobutyl, R _{7 is} not 3-chloro-4-methoxybenzyl;
if R ₁ and R ₂ combine to form an epoxy ring, R ₉ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₄ and R ₈ combine to form a didepsipeptide with structure X, R ₉ represents methyl and R ₁₀ represents isobutyl, R _{7 is} not 3-chloro-4-methoxybenzyl;
if R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ , R ₃ represents methyl, R ₅ and R ₆ combine together to form a double bond between C ₅ and C ₆ , R ₄ and R ₈ combine together the formation of a didepsipeptide with structure X, R ₉ represents methyl and R ₁₀ represents isobutyl, R _{7 is} not 3-chloro-4-methoxybenzyl; and
if R ₁ and R ₂ combine together to form an epoxy group, R ₉ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₄ binds to the terminal carboxyl group of the leucinic acid, and R ₈ binds to the terminal nitrogen of either 3-amino-2-methylpropionic acid or 3-amino-2-methylpropionic acid methyl ester, R _{7 is} not 3-chloro-4-methoxybenzyl.

The invention further provides cryptophycin compounds, where at least one of the groups attached to C ₂ , C ₈ , C ₉ , C ₁₀ and C ₁₁ has R-stereochemistry. In a further embodiment of the invention, at least one of the groups attached to C ₂ , C ₈ , C ₉ , C ₁₀ and C ₁₁ has S-stereochemistry.

где O₁ указанного X соответствует R₄ или R₅, N₈ указанного X соответствует R₈, R₉ представляет Н или низшую алкильную группу и R₁₀ представляет Н или низшую алкильную группу.The invention further provides cryptophycin compounds in accordance with the above structure, wherein the structure of a didepsipeptide that forms when R ₄ or R ₅ combines with R ₈ has the following structure X:

where O _{1 of} said X corresponds to R ₄ or R ₅ , N _{8 of} said X corresponds to R ₈ , R ₉ represents H or a lower alkyl group, and R ₁₀ represents H or a lower alkyl group.

The following terms, as used here, have the indicated meanings, unless otherwise indicated, are clearly meant to be used in context:
"lower β-amino acid" means a β-amino acid having from three to eight carbon atoms, and includes linear and non-linear hydrocarbon chains; for example 3-amino-2-methylpropionic acid.

 "Etherified lower β-amino acid" means a β-amino acid having from three to eight carbon atoms, where the hydrogen of the carboxyl group is replaced by a methyl group; for example 3-amino-2-methylpropionic acid methyl ester.

 "Lower alkanoyloxy group" means an alkanoyloxy group with one to seven carbon atoms and includes linear and non-linear hydrocarbon chains.

 "Lower α-hydroxyalkanoyloxy group" means an α-hydroxyalkanoyloxy group with two to seven carbon atoms and includes linear and non-linear hydrocarbon chains, for example, 2-hydroxy-4-methylvaleric acid.

 "Lower alkoxyl group" means any alkyl group with one to five carbon atoms bonded to an oxygen atom.

 "Lower alkyl group" means an alkyl group with one to five carbon atoms and includes linear and non-linear hydrocarbon chains including, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sesqui-butyl, methylated butyl groups, pentyl and tert-pentyl groups.

 "Allyl substituted alkene" means an alkene that contains an alkyl substituent.

 "Epoxy ring" means a three-membered ring whose skeleton consists of two carbon atoms and an oxygen atom.

 "Aziridine ring" means a three-membered ring whose skeleton consists of two carbon atoms and a nitrogen atom.

 “Episulfide ring” means a three-membered ring whose skeleton consists of two carbon atoms and a sulfur atom.

 "Sulfate ring" means a five-membered ring consisting of a carbon-carbon-oxygen-sulfur-oxygen skeleton with two additional oxygen atoms bonded to a sulfur atom.

 "Monoalkylphosphate ring" means a five-membered ring consisting of a carbon-carbon-oxygen-phosphorus-oxygen skeleton with two additional oxygen atoms, one of which contains a lower alkyl group attached to a phosphorus atom.

 “Simple unsubstituted aromatic group” refers to ordinary aromatic rings containing 4n + 2pi electrons in a monocyclic conjugated system (eg, furyl, pyrrolyl, thienyl, pyridyl) or a bicyclic conjugated system (eg, indolyl or naphthyl).

 “Simple substituted aromatic group” refers to a phenyl group substituted with a single group (eg, lower alkyl group or halogen).

 A “heteroaromatic group” refers to aromatic rings that contain one or more non-carbon substituents, such as oxygen, nitrogen, or sulfur.

"Halogen" refers to those group members in the periodic table of elements that are known as halogens. Halogenation methods include, but are not limited to, hydrogen addition, substitution at elevated temperatures, photohalogenation, etc. and such methods are known to those skilled in the art. ^{1, 2} .

Криптофицин 2
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между С₅ и С₆, R₇ представляет 4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого криптофицинового соединения, Криптофицина 4, является следующей:

Криптофицин 4
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₄ объединяются вместе с образованием тетрагидрофуранового кольца, R₂ представляет ОН группу, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₈ представляет (2-карбометоксипропил)амино группу. Структура этого соединения, Криптофицина 6, является следующей:

Криптофицин 6
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₄ объединяются вместе с образованием тетрагидрофуранового кольца, R₂ и R₈ являются ОН группами, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между С₅ и С₆ таким образом, что представляют двойную связь, R₇ представляет 3-хлор-4-метоксибензил. Структура этого соединения, Криптофицина 7, является следующей:

Криптофицин 7
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ представляет хлор, R₂ представляет ОН группу, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 8, является следующей:

Криптофицин 8
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ представляет метокси группу, R₂ представляет ОН группу, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 9, является следующей:

Криптофицин 9
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ представляет метокси группу, R₂ и R₄ являются ОН группами, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₈ представляет (2-карбоксипропил)амино группу. Структура этого соединения, Криптофицина 10, является следующей:

Криптофицин 10
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₄ объединяются вместе с образованием тетрагидрофуранового кольца, R₂ представляет ОН группу, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₈ представляет (2-карбоксипропил)амино группу. Структура этого соединения, Криптофицина 12, является следующей:

Криптофицин 12
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₄ представляет ОН группу, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₈ представляет (2-карбоксипропил)амино группу. Структура этого соединения, Криптофицина 14, является следующей:

Криптофицин 14
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-гидроксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 16, является следующей:

Криптофицин 16
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и C₆, R₇ представляет 3-хлор-4-гидроксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 17, является следующей:

Криптофицин 17
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между С₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет втор-бутил. Структура этого соединения, Криптофицина 18, является следующей:

Криптофицин 18
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀
представляет изопропил. Структура этого соединения, Криптофицина 19, является следующей:

Криптофицин 19
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидного кольца, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет водород и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 21, является следующей:

Криптофицин 21
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидного кольца, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3,5-дихлор-4-гидроксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 23, является следующей:

Криптофицин 23
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидного кольца, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и C₆, R₇ представляет 4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет водород и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 24, является следующей:

Криптофицин 24
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₄ представляет гидрокси, R₆ представляет водород, R₇ представляет 3-хлор-4-метоксибензил и R₅ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 26, является следующей:

Криптофицин 26
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет водород, R₅ и R₈ объединяются вместе с образованием двойной связи между C₅ и C₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 28, является следующей:

Криптофицин 28
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет водород, R₅ и R₈ объединяются вместе с образованием двойной связи между C₅ и C₆, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет водород и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 29, является следующей:

Криптофицин 29
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет метил, R₅ представляет гидрокси, R₆ представляет водород, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 30, является следующей:

Криптофицин 30
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3,5-дихлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 31, является следующей:

Криптофицин 31
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет метил, R₅ представляет гидрокси, R₆ представляет водород, R₇ представляет 3-хлор-4-метоксибензил и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 35, является следующей:

Криптофицин 35
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет водород, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-метоксибензил, и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 40, является следующей:

Криптофицин 40
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет водород, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3,5-дихлор-гидроксибензил, и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет изобутил. Структура этого соединения, Криптофицина 45, является следующей:

Криптофицин 45
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-метоксибензил, и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет пропил. Структура этого соединения, Криптофицина 49, является следующей:

Криптофицин 49
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₀ и С₁₁ атомами углерода, R₃ представляет водород, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и C₆, R₇ представляет 3-хлор-4-метоксибензил, и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет пропил. Структура этого соединения, Криптофицина 50, является следующей:

Криптофицин 50
Другой вариант соединения настоящего изобретения представляет соединение, где R₁ и R₂ объединяются вместе с образованием эпоксидной группы, R₃ представляет метил, R₅ и R₆ объединяются вместе с образованием двойной связи между C₅ и С₆, R₇ представляет 3-хлор-метоксибензил, и R₄ и R₈ объединяются вместе с образованием дидепсипептида со структурой X, где R₉ представляет метил и R₁₀ представляет втор-бутил. Структура этого соединения, Криптофицина 54, является следующей:

Криптофицин 54
Из вышеприведенных соединений, Криптофицины 2, 4, 16-19, 21, 23, 24, 26, 28-31, 40, 43, 45, 49, 50 и 54 являются метаболитами, продуцированными штаммом Nostoc sp. зелено-голубых водорослей (цианобактерий), которые были культивированы этими соединениями, впоследствии выделенными из этой культуры. Криптофицины 6 и 7 являются артефактами, которые продуцируются, если процедура выделения использует растворители, содержащие метанол. Криптофицины 8, 9, 10-12, 14 и 35 являются производными этих естественно продуцированных метаболитов, которые были химически модифицированы способами, описанными в экспериментальной части этой заявки, с альтернативными способами для создания приведенных в качестве примеров соединений, а также не приведенных в качестве примеров соединений, доступных для специалистов в этой области.One embodiment of the cryptophycin compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy group, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 4 -methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 2, is as follows:

Cryptophycin 2
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this cryptophycin compound, Cryptophycin 4, is as follows:

Cryptophycin 4
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₄ combine to form a tetrahydrofuran ring, R ₂ represents an OH group, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₈ represents a (2-carbomethoxypropyl) amino group. The structure of this compound, Cryptophycin 6, is as follows:

Cryptophycin 6
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₄ combine to form a tetrahydrofuran ring, R ₂ and R ₈ are OH groups, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and With ₆ so that they represent a double bond, R ₇ represents 3-chloro-4-methoxybenzyl. The structure of this compound, Cryptophycin 7, is as follows:

Cryptophycin 7
Another embodiment of the compound of the present invention is a compound wherein R ₁ is chloro, R ₂ is OH, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro- 4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 8, is as follows:

Cryptophycin 8
Another embodiment of the compound of the present invention is a compound wherein R ₁ is a methoxy group, R ₂ is an OH group, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro -4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 9, is as follows:

Cryptophycin 9
Another embodiment of the compound of the present invention is a compound wherein R ₁ is a methoxy group, R ₂ and R ₄ are OH groups, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro-4-methoxybenzyl and R ₈ represents a (2-carboxypropyl) amino group. The structure of this compound, Cryptophycin 10, is as follows:

Cryptophycin 10
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₄ combine to form a tetrahydrofuran ring, R ₂ represents an OH group, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₈ represents a (2-carboxypropyl) amino group. The structure of this compound, Cryptophycin 12, is as follows:

Cryptophycin 12
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ is methyl, R ₄ is an OH group, R ₅ and R ₆ are combined to form a double bonds between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₈ represents a (2-carboxypropyl) amino group. The structure of this compound, Cryptophycin 14, is as follows:

Cryptophycin 14
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy group, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro -4-hydroxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 16, is as follows:

Cryptophycin 16
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-hydroxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 17, is as follows:

Cryptophycin 17
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is sec-butyl. The structure of this compound, Cryptophycin 18, is as follows:

Cryptophycin 18
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents methyl, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ represents methyl and R ₁₀
represents isopropyl. The structure of this compound, Cryptophycin 19, is as follows:

Cryptophycin 19
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy ring, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro -4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is hydrogen and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 21, is as follows:

Cryptophycin 21
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy ring, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3.5 β-dichloro-4-hydroxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 23, is as follows:

Cryptophycin 23
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy ring, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is hydrogen and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 24, is as follows:

Cryptophycin 24
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ is methyl, R ₄ is hydroxy, R ₆ is hydrogen, R ₇ is 3-chloro -4-methoxybenzyl and R ₅ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 26, is as follows:

Cryptophycin 26
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents hydrogen, R ₅ and R ₈ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ represents methyl and R ₁₀ represents isobutyl. The structure of this compound, Cryptophycin 28, is as follows:

Cryptophycin 28
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents hydrogen, R ₅ and R ₈ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ represents hydrogen and R ₁₀ represents isobutyl. The structure of this compound, Cryptophycin 29, is as follows:

Cryptophycin 29
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ is methyl, R ₅ is hydroxy, R ₆ is hydrogen, R ₇ is 3-chloro -4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 30, is as follows:

Cryptophycin 30
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form an epoxy group, R ₃ represents methyl, R ₅ and R ₆ combine together to form a double bond between C ₅ and C ₆ , R ₇ represents 3.5 -dichloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 31, is as follows:

Cryptophycin 31
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy group, R ₃ is methyl, R ₅ is hydroxy, R ₆ is hydrogen, R ₇ is 3-chloro-4-methoxybenzyl and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 35, is as follows:

Cryptophycin 35
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form an epoxy group, R ₃ represents hydrogen, R ₅ and R ₆ combine together to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro -methoxybenzyl, and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is isobutyl. The structure of this compound, Cryptophycin 40, is as follows:

Cryptophycin 40
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine together to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents hydrogen, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3,5-dichloro-hydroxybenzyl, and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ represents methyl and R ₁₀ represents isobutyl. The structure of this compound, Cryptophycin 45, is as follows:

Cryptophycin 45
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy group, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro -methoxybenzyl, and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is propyl. The structure of this compound, Cryptophycin 49, is as follows:

Cryptophycin 49
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form a double bond between C ₁₀ and C ₁₁ carbon atoms, R ₃ represents hydrogen, R ₅ and R ₆ combine to form a double bond between C ₅ and C ₆ , R ₇ represents 3-chloro-4-methoxybenzyl, and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ represents methyl and R ₁₀ represents propyl. The structure of this compound, Cryptophycin 50, is as follows:

Cryptophycin 50
Another embodiment of the compound of the present invention is a compound wherein R ₁ and R ₂ combine to form an epoxy group, R ₃ is methyl, R ₅ and R ₆ are combined to form a double bond between C ₅ and C ₆ , R ₇ is 3-chloro -methoxybenzyl, and R ₄ and R ₈ combine to form a didepsipeptide with structure X, where R ₉ is methyl and R ₁₀ is sec-butyl. The structure of this compound, Cryptophycin 54, is as follows:

Cryptophycin 54
Of the above compounds, Cryptophycins 2, 4, 16-19, 21, 23, 24, 26, 28-31, 40, 43, 45, 49, 50 and 54 are metabolites produced by the strain Nostoc sp. blue-green algae (cyanobacteria), which were cultivated by these compounds, subsequently isolated from this culture. Cryptophycins 6 and 7 are artifacts that are produced if the isolation procedure uses solvents containing methanol. Cryptophycins 8, 9, 10-12, 14, and 35 are derivatives of these naturally produced metabolites that have been chemically modified by the methods described in the experimental part of this application, with alternative methods for creating exemplary compounds as well as non-exemplary compounds. compounds available to specialists in this field.

 The present invention provides methods for producing the above cryptophycin compounds by culturing a strain of Nostoc sp. Morphological characteristics of Nostoc sp. blue-green algae (cyanobacteria), as disclosed in US Pat. No. 4,946,835, are such that they are filamentous and composed of plant cells. In longer fibers, heterocysts are periodically observed in the intercalary position; Akinets are not observed. Reproduction occurs due to hormones in addition to statistical trichome breaks. Basis for identification Nostoc sp. can be found in J. Gen. Micro., 111: 1-61 (1979).

Где R₁ представляет Н, ОН, галоид, О кетонной группы, NH₂, SH, низшую алкоксильную группу или низшую алкильную группу;
R₂ представляет Н, ОН, О кетонной группы, NH₂, SH, низшую алкоксильную группу или низшую алкильную группу; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца или двойной связи между С₁₀ и С₁₁; или
R₁ и R₂ могут объединяться вместе с образованием тетрагидрофуранового кольца;
R₃ представляет Н или низшую алкильную группу;
R₄ представляют ОН, низшую алканоилокси группу или низшую α-оксиалканоилокси группу;
R₅ представляют Н или ОН группу;
R₆ представляет Н; или
R₅ и R₆ могут объединяться вместе с образованием двойной связи между C₄ и С₆;
R₇ представляет бензильную, оксибензильную, метоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидметоксибензильную или дигалоидметоксибензильную группу;
R₈ представляют ОН, низшую β-аминокислоту, где C₁ связывается с N β-аминокислоты, или этерифицированную низшую β-аминокислоту, где C₁ связывается с N этерифицированной β-аминокислотной группы;
R₄ и R₈ могут объединяться вместе с образованием дидепсипептидной группы, состоящей из низшей β-аминокислоты, связанной с низшей α-оксиалканоильной кислотой; или
R₅ и R₈ могут объединяться вместе с образованием дидепсипептидной группы, состоящей из низшей β-аминокислоты, связанной с низшей α-оксиалканоильной кислотой;
при следующих условиях:
R₁ представляет H, низшую алкильную группу или низшую алкоксильную группу только, если R₂ представляет ОН, О кетонной группы, NH₂, SH.The invention further provides that embodiment in which Nostoc sp. can be cultured and new cryptophycin metabolites, as well as previously described cryptophycin metabolites, can be isolated from this culture. In a preferred embodiment of the present invention, the strain Nostoc sp. designated GSV 224 represents a strain that is cultivated and from which compounds represented by the following structure are isolated:

Where R ₁ represents H, OH, halogen, O ketone group, NH ₂ , SH, lower alkoxyl group or lower alkyl group;
R ₂ represents H, OH, O ketone group, NH ₂ , SH, lower alkoxyl group or lower alkyl group; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, or a double bond between C ₁₀ and C ₁₁ ; or
R ₁ and R ₂ may combine to form a tetrahydrofuran ring;
R ₃ represents H or a lower alkyl group;
R ₄ represents OH, a lower alkanoyloxy group or a lower α-hydroxyalkanoyloxy group;
R ₅ represent H or OH group;
R ₆ represents H; or
R ₅ and R ₆ may combine to form a double bond between C ₄ and C ₆ ;
R ₇ represents a benzyl, oxybenzyl, methoxybenzyl, halooxybenzyl, dihalooxybenzyl, halo methoxybenzyl or dihalo methoxybenzyl group;
R ₈ represents OH, a lower β-amino acid, where C ₁ binds to N β-amino acids, or an esterified lower β-amino acid, where C ₁ binds to N an esterified β-amino acid group;
R ₄ and R ₈ can combine to form a didepsipeptide group consisting of a lower β-amino acid linked to a lower α-hydroxyalkanoyl acid; or
R ₅ and R ₈ may combine to form a didepsipeptide group consisting of a lower β-amino acid linked to a lower α-hydroxyalkanoyl acid;
under the following conditions:
R ₁ represents H, a lower alkyl group or a lower alkoxyl group only if R ₂ represents OH, O of a ketone group, NH ₂ , SH.

 In a preferred embodiment of the invention, the chemically modified cryptophycin metabolite isolated by the above method precisely provides a compound also having this structure. Methods for chemically modifying cryptophycin compounds to produce additional compounds within the scope of the present invention are available to those skilled in the art. However, additional methods are described in more detail in the experimental part of this application.

 In addition to the new cryptophycin compounds of the present invention, the present invention provides new methods for the preparation as well as the use of the above compounds, which include the following previously disclosed cryptophycin compounds, Cryptophycin 1, 3, 5, 13 and 15. The structures I of these compounds are presented at the end of the description.

 The invention is thus directed to the provision of any strain of Nostoc sp. and preferably providing a strain of Nostoc sp., GSV 224, for the production of cryptophycin compounds. To this end, GSV 224, strain Nostoc sp. , was placed in the data bank on October 7, 1993 according to the Budapest Treaty of the International Microorganism Databank for the purpose of patenting a patent culture placed in the data bank of the American Type Culture collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A. under ATCC number Acession N 55483. Other strains of Nostoc sp., in particular strain MB 5357, previously placed in the Merck and Co. database under ATCC number N 53789, represent strains that can be considered for use in the practice of the present invention.

 As is the case with other organisms, the characteristics of Nostoc sp. Are subject to change. For example, recombinants, variants, or mutants of specific strains can be obtained by treatment with various known physical or chemical mutagens, such as ultraviolet radiation, radiation, gamma radiation, or by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. All natural and induced variants, mutants and recombinants of specific strains that retain the characteristics of the production of a cryptophycin compound are included within the scope of the claimed invention.

 Cryptophycin compounds of the present invention can be obtained by culturing a strain of Nostoc sp. under aerobic conditions, immersed in an appropriate culture medium before producing mainly antibiotic activity. Other culture uses, such as surface growth on a cured medium, can also be used to produce these compounds. The culture media used for the growth of specific strains may include any medium from many nitrogen and carbon sources and inorganic salts that are known to those skilled in the art. Savings in the production process, optimal yields and ease of product isolation are those factors that are taken into account when choosing carbon sources and nitrogen sources that are used. Among the inorganic nutrient salts that can be introduced into the culture medium, conventional soluble salts are used which are capable of producing ions of iron, potassium, sodium, magnesium, calcium, ammonium, chloride, carbonate, phosphate, sulfate, nitrate and the like.

 Traces of valuable elements that are necessary for the growth and development of organisms should also be included in the cultural environment. Such traces of valuable elements are usually present in the form of impurities in other components of the medium in quantities sufficient to satisfy the growth requirements of organisms. The addition of small amounts (i.e., 0.2 ml / L) of an anti-foaming agent, such as polypropylene glycol (with a molecular weight of about 2000), may be desirable for large-scale cultivation of the medium if foaming becomes a problem.

To obtain significant amounts of cryptophycin compounds, aerobic cultivation with immersion in tanks can be used. Small amounts can be obtained using culture in a shake flask. Since the delay time in the manifestation of metabolite production is usually associated with the inoculation of large tanks by organisms, the use of a vegetative inoculum is preferred. A vegetative inoculum is prepared by inoculating a small volume of culture medium with fragments of a vegetative trichome or with an organism form containing heterocysts to produce a fresh, actively growing body culture. The vegetative inoculum is then transferred to a large tank. Wednesday,
used for the vegetative inoculum may be the same as that used for large-scale cultivations or fermentations, but another medium may be used.

Organisms can be grown at temperatures between 20 ^o C and 30 ^o C and the intensity of the incident radiation from about 100 to 200 μmol of photons m- ² sec ^-1 (photosynthetically active radiation).

As is common in processes of this type of aerobically immersed culture, gaseous carbon dioxide is introduced into the culture by adding to a sterile air stream pulsed through the culture medium. To effectively obtain cryptophycin compounds, the amount of carbon dioxide should be about 1% (at 24 ^o C and a pressure of one atmosphere).

 Previously known techniques, in particular US Pat. No. 4,946,835, provide methods for cultivating Nostoc sp., The contents of which are incorporated herein by reference.

 The production of a cryptophycin compound can occur during cultivation with test samples of the broth in relation to organisms that are known to be sensitive to these antibiotics. One suitable proven organism is Candida albicans.

 After they are obtained under the aerobic conditions of an immersed culture, the cryptophycin compounds of the invention can be isolated from the culture and from the culture medium by methods known to those skilled in the art. Isolation is usually accompanied by initially filtering the culture medium to separate the algae cells and then freeze drying the isolated cells. Freeze-dried algae can be extracted with a suitable solvent, such as ethanol, methanol, isopropanol or dichloromethane. Cryptophycins can be isolated due to the fact that this extract, as well as the culture medium, undergoes rapid chromatography on a reverse phase column. Cryptophycins can be purified by reverse phase liquid chromatography (HPLC).

 As will be apparent from the structures of cryptophycin, cryptophycin compounds contain groups that are capable of chemical modification. The parent compound of the present invention contemplates those cryptophycins that have antitumor activity. For example, derivatives exemplified in the present invention include compounds containing epoxy oxygen or hydroxyl groups on the C-7 and C-8 unit A or the leucinic acid group of unit B in Figure 1. Such derivatives of the new and previously described compounds that possess desired antitumor activity are included in the claimed invention. In addition, the relationship between the structure of cryptophycin compounds and antitumor activity is provided in the experimental part below.

 While selected cryptophycin compounds, which are known to be metabolites produced by the algae of the present invention, other cryptophycin compounds, for example Cryptophycin 8-15, can be obtained from metabolites using published techniques that are known to those skilled in the art; for example, the synthesis described in US patents 4868208, 4845086 and 4845085, the contents of which are incorporated herein by reference, or by using other methods known to specialists in this field. In addition, the present invention in the experimental part provides methods for producing derivatives.

 Cryptophycins are powerful antitumor and antifungal depsipeptides from green-blue algae (cyanobacteria) belonging to Nostocaceae. The first cryptophycin compound, Cryptophycin 1, was isolated from the land-living Nostoc sp., ATCC 53789, and was found to be very active against fungi, especially the Cryptococcus strain (RE Schwarz et al., J.lnd. Microbiol. 1990, 5: 113-124). Cryptophycin 1 has also been isolated from Nostoc sp. GSV 224, along with 24 additional algal cryptophycin analogues in the form of minimal algae constituents, and was found to be very active against subcutaneous transplanted solid tumors in mice (G. Trimurtulu et al., J. Am. Chem. Soc. 1994, 16 : 4729-4737; R. Borrow et al., J. Am. Chem. Soc. 1995, 117: 2479-2490). Two analogues from Nostoc sp. , GSV 224, Cryptophycin 3 and 5, were previously described as semi-synthetic analogues of Cryptophycin 1 (D.F. Sesin, U.S. Patent No. 4,845,085, published July 4, 1989; D.F. Sesin et al., U.S. Patent No. 4,868,208, published September 19, 1989). Cryptophycins showed significant selective cytotoxicity to the tumor in the Corbett assay and were equally cytotoxic to drug-sensitive tumor cells and drug-resistant tumor cells. Cryptophycin 1 exhibits the same form of action as vinblastine, but differs from the last drug in the irreversibility of inhibiting the microtubule ensemble (C. D. Smith et al., Cancer Res. 1994, 54: 3779-3784). One of the cryptophycins from Nostoc sp., GSV 224, Cryptophycin 24, was isolated from a sea sponge and named arenostatin A (M. Kobayashi et al., Tetrahedron Lett. 1994, 35: 7969-72; M. Kobayashi et al., Tennen Yuki Kagobutsu Toronkai Koen Yoshishu 1994, 36st, 104-110).

 Twenty-two additional cryptophycin compounds designated herein by Cryptophycin 2, 4, 6, 7, 16-19, 21, 23, 24, 26, 28-31, 40, 43, 45, 49, 50, and 54 are disclosed in Patent Application N 08/172632, filed December 21, 1993, and 08/249955, filed May 27, 1994, and International application N PCT / US94 / 14740, filed December 21, 1994, such compounds are either metabolites isolated from strain Nostoc sp., Or obtained semi-synthetic by way of such metabolites. In addition, disclosed in these patent applications is a characteristic of selected cryptophycin compounds as antimicro-tubular agents with clinical type activity manifested against a wide range of tumors implanted in mice, including DR and MDR tumors.

где Аr представляет метил, или фенил, или любую простую незамещенную, или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между C₁₈ и C₁₉;
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между С₁₃ и C₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇ , R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
X и Y каждый независимо представляют О, NH или алкиламино.The present invention provides new cryptophycin compounds having the following structure:

where Ar represents methyl, or phenyl, or any simple unsubstituted, or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

In a preferred embodiment of this cryptophycin compound, R ₈ cryptophycin is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. In another preferred embodiment of this cryptophycin compound, R ₇ is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. In a further preferred embodiment of this cryptophycin compound, R ₇ is H, R ₈ is methyl, R ₃ is methyl; X and Y are not both O.

The present invention provides an additional preferred embodiment of this cryptophycin compound, wherein R ₃ is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. In another preferred embodiment of this cryptophycin compound, R ₉ is methyl, ethyl, propyl, butyl, isobutyl, pentyl or isopentyl. In another preferred embodiment of this cryptophycin compound, R ₁₀ is methyl, ethyl, propyl, butyl, isobutyl, pentyl or isopentyl.

The invention further provides cryptophycin compounds, where at least one of the groups attached to C ₃ , C ₆ , C ₁₀ , C ₁₆ , C ₁₇ , and C ₁₈ has R-stereochemistry. In another embodiment of the invention, at least one of the groups attached to C ₃ , C ₆ , C ₁₀ , C ₁₆ , C ₁₇ , and C ₁₈ has S-stereochemistry.

КРИПТОФИЦИН 51
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃, R₇ и R₈ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 52, является следующей:

КРИФТОФИЦИН 52
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием S,S-эпоксидного кольца, R₃, R₇ и R₈ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 53, является следующей:

КРИПТОФИЦИН 53
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ представляет S-хлор, R₂ представляет R гидроксильную группу, R₃, R₇ и R₈ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 55, является следующей:

КРИПТОФИЦИН 55
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃, R₇ и R₈ представляют метил, R₄ и R₅ представляют водород, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 57, является следующей:

КРИПТОФИЦИН 57
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃, R₇ и R₈ представляют метил, R₄ и R₅ представляют водород, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 58, является следующей:

КРИПТОФИЦИН 58
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием R, R-эписульфидного кольца, R₃, R₇ и R₈ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 61, является следующей:

КРИПТОФИЦИН 61
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-метоксифенил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и C₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 81, является следующей:

КРИПТОФИЦИН 81
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет метил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и С₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 82, является следующей:

КРИПТОФИЦИН 82
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет метил, R₁ и R₂ объединяются вместе с образованием R, R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 90, является следующей:

КРИПТОФИЦИН 90
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет метил, R₁ и R₂ объединяются вместе с образованием S, S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и X и Y являются кислородом. Структура этого соединения, Криптофицина 91, является следующей:

КРИПТОФИЦИН 91
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием R, R-азиридинового кольца, R₃, R₇ и R₈ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₁₀ представляет водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 97, является следующей:

КРИПТОФИЦИН 97
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-фторфенил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 110, является следующей:

КРИПТОФИЦИН 110
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-толил, R₁ и R₂ объединяются вместе с образованием двойной связи между С₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 111, является следующей:

КРИПТОФИЦИН 111
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет 2-тиенил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 112, является следующей:

КРИПТОФИЦИН 112
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-фторфенил, R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 115, является следующей:

КРИПТОФИЦИН 115
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-фторфенил, R₁ и R₂ объединяются вместе с образованием S,S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 116, является следующей:

КРИПТОФИЦИН 116
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-толил, R₁ и R₂ объединяются вместе с образованием R, R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и C₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 117, является следующей:

КРИПТОФИЦИН 117
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-толил, R₁ и R₂ объединяются вместе с образованием S, S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и C₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 118, является следующей:

КРИПТОФИЦИН 118
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет 2-тиенил, R₁ и R₂ объединяются вместе с образованием R, R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 119, является следующей:

КРИПТОФИЦИН 119
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет 2-тиенил, R₁ и R₂ объединяются вместе с образованием S, S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y являются кислородом. Структура этого соединения, Криптофицина 120, является следующей:

КРИПТОФИЦИН 120
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и C₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, Х представляет кислород и Y представляет азот, несущий один водород. Структура этого соединения, Криптофицина 121, является следующей:

КРИПТОФИЦИН 121
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием R, R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, Х представляет кислород и Y представляет азот, несущий один водород. Структура этого соединения, Криптофицина 122, является следующей:

КРИПТОФИЦИН 122
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ и R₂ объединяются вместе с образованием S, S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, Х представляет кислород и Y представляет азот, несущий один водород. Структура этого соединения, Криптофицина 123, является следующей:

КРИПТОФИЦИН 123
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ и R₂ объединяются вместе с образованием двойной связи между C₁₈ и C₁₉, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 124, является следующей:

КРИПТОФИЦИН 124
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 125, является следующей:

КРИПТОФИЦИН 125
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ и R₂ объединяются вместе с образованием S,S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ образованием S, S-эпоксидного кольца, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 126, является следующей:

КРИПТОФИЦИН 126
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, Х представляет кислород и Y представляет азот несущий единичный водород. Структура этого соединения, Криптофицина 127, является следующей:

КРИПТОФИЦИН 127
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет фенил, R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, Х представляет кислород и Y представляет азот несущий единичный водород. Структура этого соединения, Криптофицина 127, является следующей:

КРИПТОФИЦИН 128
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-толил, R₁ представляет S-хлор, R₂ представляет S-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и C₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 130, является следующей:

КРИПТОФИЦИН 130
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-толил, R₁ представляет R-хлор, R₂ представляет R-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 131, является следующей:

КРИПТОФИЦИН 131
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между С₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 132, является следующей:

КРИПТОФИЦИН 132
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ представляет R-хлор, R₂ представляет S-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀
представляют водород, и X и Y представляют кислород. Структура этого соединения, Криптофицина 133, является следующей:

КРИПТОФИЦИН 133
Другой вариант соединения настоящего изобретения представляет соединение, когда Аr представляет п-хлорфенил, R₁ представляет R-хлор, R₂ представляет R-гидроксильную группу, R₃ и R₇ представляют метил, R₄ и R₅ объединяются вместе с образованием двойной связи между C₁₃ и С₁₄, R₆ представляет 3-хлор-4-метоксибензил, R₉ представляет изобутил, R₈ и R₁₀ представляют водород, и Х и Y представляют кислород. Структура этого соединения, Криптофицина 134, является следующей:

КРИПТОФИЦИН 134
Впредь набор, приведенный ниже представляет криптофициновые соединения, чьи замещающие группы основаны на следующей структуре:

где R₁ представляет Н или галоид;
R₂ представляет Н кислород котонной группы; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, или R₁ и R₂ могут объединяться вместе с образованием эписульфидного кольца;
R₃ представляет Н или низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ представляет Н или ОН;
R₅ представляет Н или ОН; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи;
R₆ представляет Н или галоид;
При следующем условии,
если R₁ и R₂ объединяются вместе с образованием эпоксидного кольца, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор, R₃ не является метилом. One embodiment of the cryptophycin compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 51, is as follows:
 

 Cryptophycin 51
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 52, is as follows:
 

 CRYPHOTHICINE 52
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 53, is as follows:
 

 Cryptophycin 53
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ represents S-chloro, R₂ represents R hydroxyl group, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 55, is as follows:
 

 Cryptophycin 55
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃, R₇ and R₈ represent methyl, R₄ and R₅ represent hydrogen, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 57, is as follows:
 

 Cryptophycin 57
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ represents S-chloro, R₂ represents an R-hydroxyl group, R₃, R₇ and R₈ represent methyl, R₄ and R₅ represent hydrogen, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 58, is as follows:
 

 Cryptophycin 58
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form the R, R-episulfide ring, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 61, is as follows:
 

 Cryptophycin 61
 Another embodiment of the compound of the present invention is a compound when Ar is p-methoxyphenyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 81, is as follows:
 

 Cryptophycin 81
 Another embodiment of a compound of the present invention is a compound when Ar is methyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 82, is as follows:
 

 Cryptophycin 82
 Another embodiment of a compound of the present invention is a compound when Ar is methyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 90, is as follows:
 

 Cryptophycin 90
 Another embodiment of a compound of the present invention is a compound when Ar is methyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 91, is as follows:
 

 Cryptophycin 91
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an R, R-aziridine ring, R₃, R₇ and R₈ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₁₀ represents hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 97, is as follows:
 

 Cryptophycin 97
 Another embodiment of the compound of the present invention is a compound when Ar is p-fluorophenyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 110, is as follows:
 

 Cryptophycin 110
 Another embodiment of the compound of the present invention is a compound when Ar is p-tolyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 111, is as follows:
 

 Cryptophycin 111
 Another embodiment of the compound of the present invention is a compound when Ar is 2-thienyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 112, is as follows:
 

 Cryptophycin 112
 Another embodiment of the compound of the present invention is a compound when Ar is p-fluorophenyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 115, is as follows:
 

 Cryptophycin 115
 Another embodiment of the compound of the present invention is a compound when Ar is p-fluorophenyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 116, is as follows:
 

 Cryptophycin 116
 Another embodiment of the compound of the present invention is a compound when Ar is p-tolyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 117, is as follows:
 

 Cryptophycin 117
 Another embodiment of the compound of the present invention is a compound when Ar is p-tolyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 118, is as follows:
 

 Cryptophycin 118
 Another embodiment of the compound of the present invention is a compound when Ar is 2-thienyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 119, is as follows:
 

 Cryptophycin 119
 Another embodiment of the compound of the present invention is a compound when Ar is 2-thienyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y are oxygen. The structure of this compound, Cryptophycin 120, is as follows:
 

 Cryptophycin 120
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, X represents oxygen and Y represents nitrogen carrying one hydrogen. The structure of this compound, Cryptophycin 121, is as follows:
 

 Cryptophycin 121
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, X represents oxygen and Y represents nitrogen carrying one hydrogen. The structure of this compound, Cryptophycin 122, is as follows:
 

 Cryptophycin 122
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, X represents oxygen and Y represents nitrogen carrying one hydrogen. The structure of this compound, Cryptophycin 123, is as follows:
 

 Cryptophycin 123
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ and R₂ combine together to form a double bond between C₁₈ and C₁₉, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 124, is as follows:
 

 Cryptophycin 124
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ and R₂ combine together to form an R, R-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 125, is as follows:
 

 Cryptophycin 125
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ and R₂ combine together to form an S, S-epoxy ring, R₃ and R₇ represent methyl, R₄ and R₅ the formation of S, S-epoxy rings, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 126, is as follows:
 

 Cryptophycin 126
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ represents S-chloro, R₂ represents an R-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, X represents oxygen and Y represents nitrogen carrying a single hydrogen. The structure of this compound, Cryptophycin 127, is as follows:
 

 Cryptophycin 127
 Another embodiment of a compound of the present invention is a compound when Ar is phenyl, R₁ represents S-chloro, R₂ represents an R-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, X represents oxygen and Y represents nitrogen carrying a single hydrogen. The structure of this compound, Cryptophycin 127, is as follows:
 

 Cryptophycin 128
 Another embodiment of the compound of the present invention is a compound when Ar is p-tolyl, R₁ represents S-chloro, R₂ represents an S-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 130, is as follows:
 

 Cryptophycin 130
 Another embodiment of the compound of the present invention is a compound when Ar is p-tolyl, R₁ represents R-chloro, R₂ represents an R-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 131, is as follows:
 

 Cryptophycin 131
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ represents S-chloro, R₂ represents an R-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 132, is as follows:
 

 Cryptophycin 132
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ represents R-chloro, R₂ represents an S-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀
 represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 133, is as follows:
 

 Cryptophycin 133
 Another embodiment of the compound of the present invention is a compound when Ar is p-chlorophenyl, R₁ represents R-chloro, R₂ represents an R-hydroxyl group, R₃ and R₇ represent methyl, R₄ and R₅ combine together to form a double bond between C_thirteen and C₁₄, R₆ represents 3-chloro-4-methoxybenzyl, R₉ represents isobutyl, R₈ and R₁₀ represent hydrogen, and X and Y represent oxygen. The structure of this compound, Cryptophycin 134, is as follows:
 

 Cryptophycin 134
 Henceforth, the kit below represents cryptophycin compounds whose substituent groups are based on the following structure:
 

 where r₁ represents H or halogen;
 R₂ represents H oxygen of the cotonic group; or
 R₁ and R₂ can combine to form an epoxy ring, or R₁ and R₂ can combine to form an episulfide ring;
 R₃ represents H or a lower alkyl group;
 R₄ and R₅ represent H; or
 R₄ represents H or OH;
 R₅ represents H or OH; or
 R₄ and R₅ can combine together to form a double bond;
 R₆ represents H or halogen;
 Under the following condition
 if R₁ and R₂ combine together to form an epoxy ring, R₄ and R₅ combine together to form a double bond and R₆ represents chlorine, R₃ not methyl.

R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

КРИПТОФИЦИН 20
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-бром, R₂ представляет R гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 25, является следующей:

КРИПТОФИЦИН 25
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-бром, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 27, является следующей:

КРИПТОФИЦИН 27
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃ представляет S-метил, R₄ и R₅ являются водородом и R₆ представляет хлор. Структура этого соединения, Криптофицина 32, является следующей:

КРИПТОФИЦИН 32
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁, R₄ и R₅ являются водородом, R₂ представляет S-гидроксильную группу, R₃ представляет R-метил, и R₆ представляет хлор. Структура этого соединения, Криптофицина 33, является следующей:

КРИПТОФИЦИН 33
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁, R₂, R₄ и R₅ являются водородом, R₃ представляет R-метил, и R₆ представляет водород. Структура этого соединения, Криптофицина 34, является следующей:

КРИПТОФИЦИН 34
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-бром, R₂, представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи, и R₆ представляет хлор. Структура этого соединения, Криптофицина 37, является следующей:

КРИПТОФИЦИН 37
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием S,S-эпоксидного кольца, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи, и R₆ представляет хлор. Структура этого соединения, Криптофицина 38, является следующей:

КРИПТОФИЦИН 38
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием S,R-эпоксидного кольца, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи, и R₆ представляет хлор. Структура этого соединения, Криптофицина 39, является следующей:

КРИПТОФИЦИН 39
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃ представляет S-метил, R₄ представляет S-гидроксильную группу и R₅ представляет R-гидроксильную группу и R₆ представляет хлор. Структура этого соединения, Криптофицина 41, является следующей:

КРИПТОФИЦИН 41
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием R,R-эпоксидного кольца, R₃ представляет S-метил, R₄ представляет R-гидроксильную группу и R₅ представляет S-гидроксильную группу и R₆ представляет хлор. Структура этого соединения, Криптофицина 42, является следующей:

КРИПТОФИЦИН 42
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет водород, R₂ представляет S-гидроксильную группу, R₃ представляет R-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 48, является следующей:

КРИПТОФИЦИН 48
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ представляют водород и R₆ представляет хлор. Структура этого соединения, Криптофицина 59, является следующей:

КРИПТОФИЦИН 59
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ и R₂ объединяются вместе с образованием S,S-эписульфидного кольца, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 60, является следующей:

КРИПТОФИЦИН 60
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ представляет водород, R₄ и R₅ объединяются вместе с образованием двойной связи и R₁ представляет хлор. Структура этого соединения, Криптофицина 63, является следующей:

КРИПТОФИЦИН 63
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-хлор, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ представляют водород и R₆ представляет хлор. Структура этого соединения, Криптофицина 64, является следующей:

КРИПТОФИЦИН 64
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-хлор, R₂ представляет S-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 69, является следующей:

КРИПТОФИЦИН 69
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-хлор, R₂ представляет S-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 70, является следующей:

КРИПТОФИЦИН 70
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-бром, R₂ представляет S-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 71, является следующей:

КРИПТОФИЦИН 71
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-бром, R₂ представляет S-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 72, является следующей:

КРИПТОФИЦИН 72
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-хлор, R₂ представляет S-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 73, является следующей:

КРИПТОФИЦИН 73
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-хлор, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет водород. Структура этого соединения, Криптофицина 74, является следующей:

КРИПТОФИЦИН 74
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет S-фтор, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ представляет хлор. Структура этого соединения, Криптофицина 75, является следующей:

КРИПТОФИЦИН 75
Другой вариант соединения настоящего изобретения представляет соединение, когда R₁ представляет R-фтор, R₂ представляет R-гидроксильную группу, R₃ представляет S-метил, R₄ и R₅ объединяются вместе с образованием двойной связи и R₆ - представляет хлор. Структура этого соединения, Криптофицина 76, является следующей:

КРИПТОФИЦИН 76
Настоящее изобретение обеспечивает способы получения приведенных выше криптофициновых соединений, а также ранее известных криптофицинов посредством полного синтеза.A variant of the cryptophycin compound of the present invention is a compound when R ₁ is hydrogen, R ₂ is oxygen of the ketone group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 20, is as follows:

Cryptophycin 20
Another embodiment of the compound of the present invention is a compound when R ₁ is S-bromo, R ₂ is R is a hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 25, is as follows:

Cryptophycin 25
Another embodiment of the compound of the present invention is a compound when R ₁ is R-bromo, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 27, is as follows:

Cryptophycin 27
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ is S-methyl, R ₄ and R ₅ are hydrogen and R ₆ is chloro. The structure of this compound, Cryptophycin 32, is as follows:

Cryptophycin 32
Another embodiment of the compound of the present invention is a compound when R ₁ , R ₄ and R ₅ are hydrogen, R ₂ is an S-hydroxyl group, R ₃ is R-methyl, and R ₆ is chloro. The structure of this compound, Cryptophycin 33, is as follows:

Cryptophycin 33
Another embodiment of the compound of the present invention is a compound when R ₁ , R ₂ , R ₄ and R ₅ are hydrogen, R ₃ is R-methyl, and R ₆ is hydrogen. The structure of this compound, Cryptophycin 34, is as follows:

Cryptophycin 34
Another embodiment of the compound of the present invention is a compound when R ₁ is R-bromo, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond, and R ₆ is chloro. The structure of this compound, Cryptophycin 37, is as follows:

Cryptophycin 37
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ are combined together to form an S, S-epoxy ring, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond, and R ₆ is chloro. The structure of this compound, Cryptophycin 38, is as follows:

Cryptophycin 38
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ are combined together to form an S, R-epoxy ring, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond, and R ₆ is chloro. The structure of this compound, Cryptophycin 39, is as follows:

Cryptophycin 39
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ combine to form an R, R-epoxy ring, R ₃ is S-methyl, R ₄ is an S-hydroxyl group, and R ₅ is an R-hydroxyl group and R ₆ represents chlorine. The structure of this compound, Cryptophycin 41, is as follows:

Cryptophycin 41
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ is S-methyl, R ₄ is an R-hydroxyl group and R ₅ is an S-hydroxyl group and R ₆ represents chlorine. The structure of this compound, Cryptophycin 42, is as follows:

Cryptophycin 42
Another embodiment of the compound of the present invention is a compound when R ₁ is hydrogen, R ₂ is an S-hydroxyl group, R ₃ is R-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 48, is as follows:

Cryptophycin 48
Another embodiment of the compound of the present invention is a compound when R ₁ is S-chloro, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are hydrogen, and R ₆ is chloro. The structure of this compound, Cryptophycin 59, is as follows:

Cryptophycin 59
Another embodiment of the compound of the present invention is a compound when R ₁ and R ₂ are combined to form an S, S-episulfide ring, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 60, is as follows:

Cryptophycin 60
Another embodiment of the compound of the present invention is a compound when R ₁ is S-chloro, R ₂ is an R-hydroxyl group, R ₃ is hydrogen, R ₄ and R ₅ are combined to form a double bond and R ₁ is chloro. The structure of this compound, Cryptophycin 63, is as follows:

Cryptophycin 63
Another embodiment of the compound of the present invention is a compound when R ₁ is R-chloro, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are hydrogen and R ₆ is chloro. The structure of this compound, Cryptophycin 64, is as follows:

Cryptophycin 64
Another embodiment of the compound of the present invention is a compound when R ₁ is R-chloro, R ₂ is an S-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 69, is as follows:

Cryptophycin 69
Another embodiment of the compound of the present invention is a compound when R ₁ is S-chloro, R ₂ is an S-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 70, is as follows:

Cryptophycin 70
Another embodiment of the compound of the present invention is a compound when R ₁ is R-bromo, R ₂ is an S-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 71, is as follows:

Cryptophycin 71
Another embodiment of the compound of the present invention is a compound when R ₁ is S-bromo, R ₂ is an S-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 72, is as follows:

Cryptophycin 72
Another embodiment of the compound of the present invention is a compound when R ₁ is S-chloro, R ₂ is an S-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 73, is as follows:

Cryptophycin 73
Another embodiment of the compound of the present invention is a compound when R ₁ is S-chloro, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is hydrogen. The structure of this compound, Cryptophycin 74, is as follows:

Cryptophycin 74
Another embodiment of the compound of the present invention is a compound when R ₁ is S-fluoro, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 75, is as follows:

Cryptophycin 75
Another embodiment of the compound of the present invention is a compound when R ₁ is R-fluoro, R ₂ is an R-hydroxyl group, R ₃ is S-methyl, R ₄ and R ₅ are combined to form a double bond and R ₆ is chloro. The structure of this compound, Cryptophycin 76, is as follows:

Cryptophycin 76
The present invention provides methods for producing the above cryptophycin compounds, as well as previously known cryptophycin by complete synthesis.

 The invention further provides that new cryptophycin metabolites, as well as previously described cryptophycin metabolites, can be synthesized using the methods provided in this invention.

-аминокислотой с образованием второй субединицы; и сочетание первой субединицы со второй субединицей с образованием криптофицина.The present invention provides a method for producing a cryptophycin compound comprising selective allyl substituted E alkene; rearrangement of allyl substituted E alkene via Wittig stereospecific rearrangement; the conversion of this compound to the first δ-amino acid or δ-hydroxy acid, a combination of the first acid with the second α-amino acid to form the first subunit; a combination of a third β-amino acid with a fourth α-hydroxy acid or

-amino acid with the formation of the second subunit; and combining the first subunit with the second subunit to form cryptophycin.

где Аr представляет метил или фенил или любую простую незамещенную или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между C₁₈ и C₁₉;
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между С₁₃ и С₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇, R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
Х и Y каждый независимо представляют О, NH или алкиламино.The present invention further provides a preferred embodiment of the method, where the obtained cryptophycin compound has the following structure:

where Ar represents methyl or phenyl or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

In a preferred embodiment of the present invention, the method provides a cryptophycin compound, wherein Ar is phenyl, R ₃ is methyl and R ₆ is halo methoxybenzyl, R ₇ is H; R ₈ represents methyl; R ₉ represents isobutyl; R ₁₀ represents H; X represents O; and Y represents O.

где R₁ представляет галоид; R₂ представляет ОН; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца;
R₃ представляет Н; и R₄ представляют Н; или
R₃ и R₄ могут объединяться вместе с образованием двойной связи.In addition to the fact that the present invention provides cryptophycin compounds with the above structure, the present invention provides methods for producing previously disclosed cryptophycin compounds and cryptophycin compounds known in the art. Cryptophycin compounds 1, 8 and 35 were obtained using complete synthesis. The following compound is an example of previously disclosed and previously known in the writer cryptophycin compounds obtained by complete synthesis:

where R ₁ represents a halogen; R ₂ represents OH; or
R ₁ and R ₂ may combine to form an epoxy ring;
R ₃ represents H; and R ₄ represent H; or
R ₃ and R ₄ may combine to form a double bond.

где Аr представляет метил или фенил или любую простую незамещенную или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₁ могут объединяться вместе с образованием двойной связи между C₁₈ и C₁₉;
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между C₁₃ и C₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇, R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
Х и Y каждый независимо представляют О, NH или алкиламино;
вместе с фармацевтически приемлемым носителем.The present invention also provides a pharmaceutical composition useful for inhibiting the proliferation of a mammalian hyperproliferative cell, comprising an effective amount of a cryptophycin compound with the following structure:

where Ar represents methyl or phenyl or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₁ may combine to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino;
together with a pharmaceutically acceptable carrier.

 In a preferred embodiment of the present invention, the pharmaceutical composition further includes at least one additional antitumor agent.

где Аr представляет метил, или фенил, или любую простую незамещенную или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между С₁₈ и C₁₉,
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между С₁₃ и C₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇, R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
Х и Y каждый независимо представляют О, NH или алкиламино.The present invention also provides a method for inhibiting mammalian cell proliferation, comprising contacting the mammalian cell with a cryptophycin compound in an amount sufficient to inhibit cell proliferation, a cryptophycin compound having the following structure:

where Ar represents methyl, or phenyl, or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ ,
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

 In a preferred embodiment of the present invention, this method further comprises contacting the cell with at least one additional antitumor agent. In a preferred embodiment of the present invention, the mammalian cell that is being contacted is hyperproliferative. In another preferred embodiment of the present invention, the hyperproliferative cell is human.

где Ar представляет метил, или фенил, или любую простую незамещенную или замещенную ароматическую или гетероароматическую группу;
R₁ представляет галоид, SH, амино, моноалкиламино, диалкиламино, триалкиламмоний, алкилтио, диалкилсульфоний, сульфат или фосфат;
R₂ представляет ОН или SH; или
R₁ и R₂ могут объединяться вместе с образованием эпоксидного кольца, азиридинового кольца, эписульфидного кольца, сульфатного кольца или моноалкилфосфатного кольца; или
R₁ и R₂ могут объединяться вместе с образованием двойной связи между С₁₈ и C₁₉;
R₃ представляет низшую алкильную группу;
R₄ и R₅ представляют Н; или
R₄ и R₅ могут объединяться вместе с образованием двойной связи между C₁₃ и С₁₄;
R₆ представляет бензильную, оксибензильную, алкоксибензильную, галоидоксибензильную, дигалоидоксибензильную, галоидалкоксибензильную или дигалоидалкоксибензильную группу;
R₇, R₈, R₉ и R₁₀ каждый независимо представляют Н или низшую алкильную группу; и
Х и Y каждый независимо представляют О, NH или алкиламино.The present invention also provides a method of inhibiting the proliferation of a mammalian hyperproliferative cell having a plurality of drug resistance phenotypes, a method comprising contacting the cell with an amount of a cryptophycin compound effective to disrupt the dynamic state of microtubule polymerization and depolymerization to delay cell mitosis, thereby inhibiting cell proliferation, a cryptophycin compound having the following structure:

where Ar represents methyl, or phenyl, or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen, SH, amino, monoalkylamino, dialkylamino, trialkylammonium, alkylthio, dialkylsulfonium, sulfate or phosphate;
R ₂ represents OH or SH; or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring, an episulfide ring, a sulfate ring, or a monoalkyl phosphate ring; or
R ₁ and R ₂ may combine together to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl, alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and
X and Y each independently represent O, NH or alkylamino.

 In a preferred embodiment of the present invention, the method further comprises contacting the cell with at least one additional antitumor agent. In another preferred embodiment of the present invention, the mammalian cell is human.

 The present invention also provides a method of alleviating a pathological condition caused by hyperproliferative mammalian cells, a method comprising administering to a subject an effective amount of a pharmaceutical composition disclosed herein to inhibit cell proliferation. In a preferred embodiment of the present invention, the mammalian cells are human.

 In a preferred embodiment of the present invention, the method further comprises conducting the subject at least one additional therapy aimed at alleviating the pathological condition. In a preferred embodiment of the present invention, the pathological condition is characterized by the formation of tumors. In another preferred embodiment of the present invention, the tumors are selected from the group consisting of breast, small chamber lung, non-small chamber lung, calorectal leukemia, melanoma, pancreatic adenocarcinoma, central nervous system (CNS), ovarian prostate, sarcoma of soft tissue or bone, head and neck, gastritis, which includes pancreatic gastritis and gastric gastritis, stomach, myeloma, bladder, renal neuroendocrinia, which includes the thyroid gland and tumors that are not associated with the disease m Hodgkin and cancer, Hodgkin's disease-related.

 The method for preparing cryptophycin compounds is summarized in Scheme 1, as shown in Figure 5. The starting material is the 3-alkene (a) substituted at the C-2 position by an XH group in the S-configuration, where X is oxygen or NH. L-alanine and L-lactic acid serve as an inexpensive source of starting material a. A key step in the synthesis is the stereo-selective [2, 3] Wittig rearrangement (DJ - S. Tsai et al., J. Org. Chem., 1984, 49: 1842-1843; K. Mikami et al., Tetrahedron 1984, 25: 2303-2308; N. Sayo et al., Chem. Lett., 1984, 259-262) of a (b) propargyl ether in (3R, 4R) -3- (XH substituted) -4-alkylhept-5 (E ) -en-1-yn (c), where X is oxygen or protected nitrogen (e.g. tert-butyldimethylsilylamino). Compound c can then be converted to the δ-hydroxy or amino acid link A of the cryptophycin precursor, methyl (5S, 6R) -5- (XP-substituted) -6-alkyl-8-aryl-octa-2E, 7E-dienoate (d ), where P is a suitable protecting group using methods known to those skilled in the art.

 One of the strategies for synthesizing a cryptophycin compound, which is composed of a δ-hydroxy or amino acid link A, an α-amino acid link B, a β-amino acid link C and an α-hydroxy or amino acid link D, is to create a macrocycle unit from two precursors representing residues of the cryptophycin molecule for example, AB of the precursor (e) containing the δ-hydroxy or amino acid link A and the α-amino acid link B and CD of the precursor (f) containing the β-amino acid link C and α-hydroxy or the amino acid link D.

 In the method described herein, the cryptophycin compound is composed of AB and C-D precursors in two stages by (1) connecting the end units A and D to AB and C-D precursors to form a cyclic product.

 In the synthesis of Cryptophycin 51 described in the experimental part, an ether bond is formed between the δ-hydroxy group of unit A in the AB residue and the carboxylic acid group of unit D in the CD fragment to form an acyclic C-DAB intermediate and then the amide bond is formed between the carboxylic acid group unit B in AB residue and β-amino group of unit C in the CD residue. Compound K is a precursor of an AB residue, compound P is a precursor of a CD residue, and compound R is an acyclic precursor of Cryptophycin 51 CDAB. Compounds K and P contain protecting groups on a carboxylic acid group of unit B and a β-amino group of unit C to limit coupling with the formation of ether between units A and D in stage 1. These protecting groups are removed from the CDAB intermediate so that amide formation can occur between units B and C in stage 2.

The synthesis of methyl (5S, 6R) -5-tert-butyldimethylsilyloxy-6-methyl-8-phenyl-octa-2E, 7E-dienoate (G), link A of the cryptophycin precursor 51 is summarized in Scheme 2 (Figure 6). (S) trans-3-penten-2-ol (A), the starting material was prepared by enzymatic cleavage of the racemic compound. Reaction of A with propargyl chloride and base under phase transfer conditions gives propargyl ether B in 86% yield. Processing B with butyl lithium at -90 ^° C leads to alcohol C with 71% yield. The desired 3R, 4R anti-compound C was a product obtained only by Wittig rearrangement. After protecting the hydroxyl group C in the form of tert-butyldimethylsilyl ether (or tert-butyldimethylsilyl ether), hydroboration of the triple bond (N.C. Brown, Organic Synthesis Via Boranes, Willey, 1975) leads to aldehyde D with a 73% yield from C. Then D was converted to trans α, β-unsaturated ester E by Horner-Emons reaction in 90% yield. Selective ozonolysis of the C6-C7 double bond in D gives the aldehyde F in 83% yield. Finally, the reaction of Wittig F with benzyl triphenylphosphonium chloride in the presence of butyllithium gives G in 80% yield. The yield of G from A was 26%.

The reaction of combining unit A of the precursor Q with D-3- (3-chloro-4-methoxyphenyl) alanine of unit B to form the AB precursor (K) is summarized in Scheme 3 (Figure 7). Hydrolysis of the methyl ester group in G with lithium hydroxide in acetone gives carboxylic acid H in 95% yield. The combination of H with trichloroethyl ether I to form J can be achieved in 65% yield by treating a solution of H in N, N-dimethylformamide (DMF) with a small excess of pentafluorophenyl diphenylphosphinate (FDPP), an equimolar amount of trifluoroacetate salt I, followed by treatment with 3 equivalents of diamine of diisopropyl DIEA) at 25 ^° C (S. Chen et al., Tetrahedron Lett. 1991, 32: 6711-6714). Fluorodesilylation J then leads to K in 95% yield.

 Protected amino acid I was prepared from D-tyrosine in five stages. Initially, D-tyrosine was chlorinated with sulfuryl chloride in glacial acetic acid (R. Zeynek, Hoppe-Seyler's Z.f. Physiol. Chemie 1926, 144: 247-254). Then N- (tert-butoxycarbonyl) -3- (3-chlorop-4-hydroxyphenyl) -D-alanine was obtained in 94% yield by treating the suspension of the amino acid in a 50% aqueous solution of dioxane with ditret-butyl dicarbonate in the presence of triethylamine. The resulting product was demethylated with dimethyl sulfate in the presence of potassium carbonate by refluxing in acetone in 84% yield. Then, methyl ester was saponified with sodium hydroxide in an aqueous solution of dioxane to form N- (tert-butoxycarbonyl) -3- (3-chloro-4-methoxyphenyl) -D-alanine in 86% yield. The aging of the BOC-protected amino acid with trichloroethanol, pyridine and DDC in dichloromethane leads to trichlorethyl ether I in 65% yield. Treatment of this substance with trifluoroacetic acid (TFA) results in a quantitative yield of trifluoroacetate salt I.

 The synthesis of (2S) -2- [3 '(tert-butoxycarbonyl) amino-2', 2'-dimethylpropanoyloxy] -4-methylpentanoic acid (P), a C-D precursor, is summarized in Scheme 4 (Figure 8). The starting point for unit C of part P was amino alcohol L. Protection of the amino group in L by treatment with di-tert-butyl carbonate in the presence of triethylamine (in 93% yield), followed by oxidation of the primary alcohol with rhenium tetroxide (PHJ, Cerlsen et al., J Org. Chem. 1981, 46: 3936-3938) gives carboxylic acid M (in 66% yield). L-leucinic acid was converted to allyl ether N in 93% yield under phase transfer conditions by keeping it in a mixture of allyl bromide in dichloromethane and an aqueous solution of sodium bicarbonate containing tetra-n-butylammonium chloride (S. Friedrich-Bochnitschek et al. , J. Org. Chem., 1989, 54: 751-756). The reaction of combining M with N was carried out with 4-dimethylaminopyridine (DMAP) and dichlorocyclocarbodiimide (DCC) in dichloromethane to give O in 75% yield. The allyl ether O was cleaved in THF containing morpholine and a tetrakis (triphenylphosphine) -palladium catalyst with the formation of P in 95% yield (P. D. Jeffrey et al., J. Org. Chem. 1982, 47: 587-590).

 The combination of AB precursor (K) and C-D precursor (P) was performed as shown in scheme 5 (figure 9). Treatment of K and P with DCC / DMAP in dichloromethane affords the fully protected C-D-A-B intermediate (Q) in 84% yield. Reductive cleavage of the trichloroethyl ether group in Q was achieved using activated zinc dust in acetic acid. The BOC-protected group was then removed with trifluoroacetic acid to give R as the trifluoroacetate salt in 91% of the total yield from Q.

 Macrolactamization of R using FDPP leads to Cryptophycin 51 with 61% yield (J. Dudash, Jr.et al., Synth.Commun, 1993, 23: 349-356). The total yield from S-trans-3-penten-2-ol (A) was 7%.

 Cryptophycin 51 served as a precursor of Cryptophycin 52, R, R-epoxide, and Cryptophycin 53 S, S-epoxide. In turn, Cryptophycin 52 served as a precursor to Cryptophycin 55, 18R, 19S-chlorohydrin and Cryptophycin 57, 13,14-dihydro analog. Cryptophycin 57 served as a precursor of Cryptophycin 58. Cryptophycin 53 served as a precursor of Cryptophycin 61 using the method described by T.H. Chan and J.F. Finkenbine, J. Am. Chem. Soc., 1972, 94: 2880-2882; and Cryptophycin 97 using the method described by Y. Ittah et al., J. Org. Chem., 1978, 43: 4271-4273.

For the synthesis of cryptophycins that contained Ar groups that differ from phenyl, the A link of the precursors of the general formula d (Scheme 1, R ₃ = Me) can be prepared by the Wittig reaction of the aldehyde F (Scheme 2; TBS-protecting group) or S (Scheme 6 ; TBPS protecting group) with the corresponding aryl triphenylphosphonium chloride in the presence of butyllithium. Cryptophycin 81 was prepared from the precursor d (Ar = p-methoxyphenyl, R ₃ = Me) as shown in schemes 6 and 7 (figures 10 and 11).

The ar group can also be introduced into a new cryptophycin at a later stage of the synthesis. First, the precursor d (Ar = R ₃ = Me) was converted to Cryptophycin 82 by a combination of the corresponding AB (e) and CD (f) precursors, as shown in Scheme 8 (Figure 12). Selective ozonolysis of Cryptophycin 82 or periodic acid oxidation of the corresponding cryptophycin epoxides 90 and 91 leads to Cryptophycin aldehyde 108. The reaction of Wittig Cryptophycin 108 with the corresponding aryl triphenylphosphonium chloride in the presence of butyllithium gives a new cryptophycin (Scheme 9; Figure 13). Using this procedure, Cryptophycin 110 (Ar = p-fluorophenyl), Cryptophycin 111 (Ar = p-tolyl), Cryptophycin 112 (Ar = 2-thienyl) and Cryptophycin 124 (Ar = p-chlorophenyl) were obtained. Cryptophycin 110 served as a precursor of Cryptophycin 115 and 116 epoxides. Cryptophycin 111 served as a precursor of Cryptophycin 117 and 118 epoxides and Cryptophycin chlorhydrins 128, 130 and 131. Cryptophycin 112 served as a precursor of Cryptophycin epoxides 119 and 120. Cryptophycin Cryptophycin epoxides 125, 126 and Cryptophycin chlorohydrins 132, 133 and 134.

 Another strategy in the synthesis of cryptophycin is the creation of a macrocycle assembly of three precursors, for example, AB of the precursor (e) containing a δ-hydroxy or amino acid unit A, a precursor containing a δ-hydroxy or amino acid unit D, and a precursor containing α-amino acid C link. In the method described here, cryptophycin comprises a node of AB, C and D precursors in three stages due to (1) the connection of terminal A and D in AB and D precursors with the formation of an acyclic DAB intermediate, (2) end link connections D C. D-A-B and C precursors to form an acyclic C-D-A-B intermediate, and (3) Connection terminal units B and C to form the cyclic product.

 In the synthesis of cryptophycin 121 described in the experimental part, an ether bond is formed between the δ-hydroxy group of unit A in the AB residue and the carboxylic acid group of unit D in the D fragment to form an acyclic D-A-B intermediate. An amide bond is then formed between the carboxylic acid group of unit C and the α-amino group of unit D in the D-A-B fragment. Finally, an amide bond is formed between the carboxylic acid group of unit B in the AB residue and the β-amino group of unit C in the C-D residue. Compound K is a precursor with an AB residue, BOC-L-leucinaldehyde is a precursor of D and compound AL is a precursor of C. Compound AK is an acyclic C-D-A-B precursor of Cryptophycin 121 (Scheme 10; Figure 14). Compounds K and BOC-L-leucinaldehyde contain protecting groups on the carboxylic acid group of unit B and the α-amino group of unit D to limit the coupling reaction to form an ester between units A and D in step 1. The protecting group is removed from the δ-amino group of unit D in DAB intermediate, so that amide formation can occur between amino group C and carboxylic acid C in stage 2. These protecting groups are removed from the CDAB intermediate so that amide formation can occur between units B and C in stage 2.

 The new cryptophycin compounds of the present invention contain epoxy rings that open with nucleophiles at different speeds than the cryptophycin 1 epoxy ring, or contain chlorohydrin functional groups that form epoxy rings at different speeds than the cryptophycin 8 chlorohydrin functional groups, turning into the cryptophycin epoxy ring 1. Cryptophycin 1 epoxy ring or chlorohydrin functional group (masked epoxy ring) Cryptophycin 8 is a su ety for optimal activity in vivo. If epoxy oxygen is removed (as found in Cryptophycin 3) or the epoxy ring is hydrolyzed to a diol (as found in Cryptophycin 15), the antitumor activity is greatly reduced. Cryptophycin 1 exhibits noticeable toxicity in animals, compared to Cryptophycin 8. This is reflected in terms of T / C (mostly above 0%) and the total values of Log destruction (mostly below 2.0) for Cryptophycin 1, compared with these values for Cryptophycin 8 (for the most part T / C values of 0% and total values of Log destruction above 2.8). Cryptophycin 25, corresponding to the bromohydrin analogue, shows the T / C ratio and the total values of Log destruction, which are comparable to those for Cryptophycin 1. This striking difference in in vivo activity confirms that Cryptophycin 25 bromohydrin converts more quickly to Cryptophycin 1 in vivo, than Cryptophycin 8. This is further confirmation that the less toxic Cryptophycin 8 may have more time to accumulate in the tumor area before becoming the active compound of Cryptophycin 1. Epoxy group and cryptophycin 1 probably binds covalently to its target receptor in the tumor cell. The new cryptophycins in the present invention may potentially have better in vivo activity than Cryptophycin 1 and Cryptophycin 8 due to the manifestation of more favorable in vivo epoxide formation rates from the corresponding chlorohydrin prodrugs and covalent binding to the target receptor in the tumor cell.

 The compounds of the present invention are more resistant to hydrolysis and solvolysis than Cryptophycin 1 and 21. The ester bond connecting the C and D units in Cryptophycin 1 is relatively sensitive to mild basic hydrolysis, cleaving at pH 11 to hydroxy acid with a half-life of 0.83 hours. The C-D ether bond in Cryptophycin 21, which lacks the C-2 methyl group on the C atom, opens up faster with a half-life of 0.25 hours. The C-D ester bond is also sensitive to solvolysis. If methanol is used in the isolation scheme, significant methanolysis of Cryptophycin 1 and 21 takes place. Cryptophycin 21 is much more susceptible to methanolysis than Cryptophycin 1. Cryptophycin 1 exhibits antitumor activity, while Cryptophycin 21 is inactive, probably because the CD ester bond Cryptophycin 21 is hydrolyzed faster than the cryptophycin 1 CD ester bond in vivo. By hydrolysis of the C-D ether bond, a partial decrease in the in vivo activity of Cryptophycin 1 during the intraperitoneal and subcutaneous routes of administration of the drug can also be explained. The C-D ester bond of cryptophycins containing two methyl groups on the C-2 atom of C, such as that found in Cryptophycin 52, is stable at pH 11.

The compounds of the present invention and previously discovered cryptophycin compounds can be used therapeutically as antitumor agents and thereby be used in methods for treating tumor diseases. The term "neoplastic", as used here, refers to a tumor that undergoes abnormal growth, such growth that occurs due to the proliferation of cells that are not subject to the usual growth restrictions. The term "anti-neoplastic agent" (antitumor agent), as used here, refers to any compound, composition, mixture, joint mixture or combination that inhibit, remove, slow down or change the neoplastic cell phenotype,
Chemotherapy, surgery, radiation therapy, therapy with biological response modifiers and immunotherapy are currently used in the treatment of cancer. Each type of therapy has specific indications that are known to specialists in this field, and one of the types of therapy or all of them can be used to achieve complete destruction of neoplastic cells. Chemotherapy using one or more cryptophycins is provided by the present invention. In addition, a combination of chemotherapy processes, chemotherapy using cryptophycins in combination with other neoplastic agents, is also provided by the subject invention in the form of combination therapy, which is usually more effective than using single neoplastic agents. Thus, a further aspect of the present invention provides compositions comprising a therapeutically effective amount of at least one new cryptophycin compound of the present invention, including non-toxic addition salts thereof, which serve to provide the above therapeutic beneficial effect. Such compositions may also be provided with a physiologically suitable liquid, gel or solid carriers, diluents, adjuvants and excipients. Such carriers, diluents, adjuvants and excipients may be found in United States Pharmacopeia Vol. XXII and National Formulary Vol. Xvii. US Pharmacopeia Convention, Inc., Rockville, MD (1989), the contents of which are incorporated herein by reference. Additional forms of processing are provided in AHFS Drug Information, 1993 ed. by American Hospital Formulary Service, pp. 522-660, the contents of which are incorporated herein by reference.

 The present invention further provides that the pharmaceutical composition used to treat a neoplastic disease comprises at least one cryptophycin compound and at least one additional antitumor agent. Antitumor compounds that can be used in combination with cryptophycin include those provided by The Merck Index. 11th ed. Merck and Co., Inc. (1989) pp. Ther 16-17, the contents of which are incorporated herein by reference. In another embodiment of the invention, the antitumor agents may be antimetabolites, which may include, but are not limited to, methotrixan, 5-fluorouracil, 6-mercaptopurine, cytosine arabinoside, hydroxyurea, and 2-chlorodeoxyadenosine. In another embodiment of the present invention, antitumor agents contemplate alkylating agents, which may include, but are not limited to, cyclophosphamide, melphalan, busulfan, paraplatin, chlorambucil and nitrogen mustard. In another embodiment of the invention, the antitumor agents are plant alkaloids, which may include, but are not limited to, vincristine, vinblastine, taxol, and etopside. In another embodiment of the present invention, the antitumor agents contemplated are antibiotics, which may include, but are not limited to, doxorubicin (adriamycin), daunorubicin, mitomycin c, and bleomycin. In another embodiment, the antitumor agents contemplated are harmones, which may include, but are not limited to, calousterone, diomestavolone, proiponate, epithiostanol, mepitiostan, testolactone, tamoxifen, polyestradiol phosphate, mesterol acetate, flutamide, nilutamide and trilotane. In another aspect of the invention, contemplated antitumor agents include enzymes that may include, but are not limited to, L-Asparaginase or aminoacridine derivatives, which may include, but are not limited to, amsacrine. Additional antitumor agents include those provided by Skeel, Roland T., “Antitumor drugs and biological response modifier: classification, use and toxicity of clinically beneficial agents,” Handbook of Cancer Chemotherapy (3rd ed.), Little Brown and Co. (1991), the contents of which are incorporated herein by reference.

 The cryptophycin compounds of the present invention and compositions can be administered to mammals for veterinary use, such as domestic animals, and for clinical use in humans, in a manner analogous to other therapeutic agents. Basically, the dose required for therapeutic efficacy will vary according to the type of use and form of administration, as well as the specific requirements of the individual hosts. Typically, doses will be in the range of about 0.001 to 1000 mg / kg, more typically 0.01 to 10 mg / kg of the host body weight. Alternatively, doses within these areas can be administered by continuous infusion over a longer period of time, usually in excess of 24 hours, until the desired therapeutic effect is achieved. Indeed, the drug dose as well as the route of administration should be selected based on relative efficacy, relative toxicity, tumor growth characteristics and the effect of cryptophycin on the cell cycle, drug pharmacokinetic characteristics, age, gender, physical condition of the patient and previous treatment.

 Cryptophycin compounds with or without additional antitumor agents can be formulated in therapeutic formulations in their natural form or in salt form. Pharmaceutically acceptable, non-toxic salts include basic addition salts (prepared with free carboxylic or other anionic groups) which can be prepared from inorganic bases such as, for example, sodium, potassium, ammonium, calcium or iron hydroxides, and organic bases such as isopropylamine , trimethylamine, 2-ethylaminoethanol, histidine, procaine and the like. Such salts can also be obtained as acid addition salts with any free cationic groups and will usually form with inorganic acids, such as, for example, hydrochloric or phosphoric acids, or organic acids, such as acetic, oxalic, tartaric, mandelic acid and the like. like. Additional excipients that the invention further provides are those excipients that are available to those skilled in the art, for example, those found in United States Pharmacopeia Vol. XXII and National Formulary Vol.XVII, U.S. Pharmacopeia Convention, Inc., Rockville, MD (1989), the contents of which are incorporated herein by reference.

 The suitability of a particular carrier for inclusion in a given therapeutic composition depends on the preferred route of administration. For example, antitumor compositions may be formulated for oral administration. Such compositions are usually prepared either in the form of a liquid solution or in the form of suspensions or in the form of solid forms. Oral compositions typically include such commonly used additives as binders, fillers, carriers, preservatives, stabilizing agents, emulsifiers, buffers and fillers, such as, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate and the like. These compositions are prepared in the form of solutions, suspensions, tablets, pills, capsules, subject to release of the compositions, or powders, and usually contain 1-95% of the active ingredient, preferably 2-70%.

 The compositions of the present invention can also be prepared in the form of injectable, or in the form of liquid solutions, suspensions, or emulsions; solid forms suitable for dissolution in, suspension in, or may be prepared in liquid form prior to injection. Such injections can be administered subcutaneously, intravenously, intraperitoneally, intramuscularly, intratracheally or intrapleurally. The active ingredient or ingredients are often mixed with diluents or excipients that are physiologically tolerant or compatible with the active ingredient (s). Suitable diluents and fillers are, for example, water, saline, dextrose, glycerin or the like, and combinations thereof. In addition, if desired, the compositions may contain minimal amounts of auxiliary substances, such as wetting or emulsifying agents, stabilizing or pH buffering agents.

The invention further provides methods for using cryptophycin compounds, combined by a common type of structure, to inhibit the proliferation of mammalian cells by contacting these cells with the cryptophycin compound in an amount sufficient to inhibit the proliferation of mammalian cells. A preferred embodiment is a method of inhibiting the proliferation of hyperproliferative mammalian cells. For the purposes of this invention, “mammalian hyperproliferative cells” are mammalian cells that do not undergo characteristic growth restrictions, such as programmed cell death (apoptosis). Another preferred embodiment is when the mammalian cell is human. The invention further provides contacting a mammalian cell with at least one cryptophycin compound and at least one additional antitumor agent. The types of antitumor agents considered are the same as those disclosed hereinabove,
The invention further provides methods of using cryptophycin compounds, combined by a common type of structure, to inhibit the proliferation of hyperproliferative cells with drug resistance phenotypes, including compounds with a variety of drug resistance phenotypes, by contacting the indicated cell with a cryptophycin compound in an amount sufficient to inhibit the proliferation of mammalian hyperproliferative cells. A preferred embodiment is when the mammalian cell is human. The invention further provides contact of a mammalian cell with a cryptophycin compound and at least one additional antitumor agent. The types of antitumor agents considered are those such as those disclosed hereinabove.

 The invention further provides a method for alleviating pathological conditions caused by hyperproliferating mammalian cells, for example, neoplasia, by administering to the subject an effective amount of the pharmaceutical composition provided herein above to inhibit the proliferation of hyperproliferating cells. The term "pathological condition", as used here, refers to any pathology that occurs during the proliferation of mammalian cells that are not subject to normal cell growth restrictions. Such cell proliferation may be due to tumors, including, but not limited to, the following tumors: breast, small chamber, non-small chamber, calorectal leukemia, melanoma, pancreatic adenocarcinoma, central nervous system (CNS), ovarian prostate, soft tissue or bone sarcoma , head and neck, gastritis, which includes pancreatic gastritis and gastric gastritis, stomach, myeloma, bladder, renal neuroendocrinia, which includes the thyroid gland and neoplasms, not associated with Hodkins disease, and neoplasms associated with Hodkins disease. In another embodiment of the invention, the tumor cells are human. The present invention further provides methods of alleviating such pathological conditions using cryptophycin in combination with other therapy, as well as with other antitumor agents. Such therapies and their correspondence to various tumors can be found in Cancer Principles and Practice of Oncology. 4th.ed /, Editors De Vita, V., Hellmann, S. and Rosenberg, S., Uppincott Co. (1993), the contents of which are incorporated herein by reference.

 In the present invention, cryptophycin compounds are manifested in a highly disturbed microtubule structure in cultured cells. In addition, in contrast to Vinca alkaloids, cryptophycin compounds are obviously poor substrates for drug outflow from P-glycoprotein. Cryptophycin 1 is the most cytotoxic for the strain of blue-green algae (cyanobacteria) Nostoc sp., Designated as GSV 224 and exhibits excellent activity against tumors implanted in mice. This cyclic didepsipeptide was previously isolated from Nostoc sp. ATCC N 53789 as an antifungal agent and its complete structure has been previously determined. The relative and absolute stereochemistry of this potentially important drug has been established using a combination of chemical and spectral methods. Twenty-four additional cryptophycin compounds, Cryptophycins 2-7, 16-19, 21, 23, 24, 26, 28-31, 40, 43, 45, 49, 50 and 54, were also isolated from GSV 224 and their complete structures were determined and cytotoxicity. Some derivatives and degradation products are described from a chemical and pharmaceutical point of view.

 The following examples serve to illustrate certain preferred variants and aspects of the present invention and are not considered as limiting the scope of the invention.

 Experimental part.

 In the description of the experimental part that follows, all weights are given in grams (g), milligrams (mg), micrograms (μg), nanograms (ng), picograms (pkg), moles (mol) or millimoles (mmol), all concentrations are given as volume percent (%), molar (M), millimolar (mm), micromolar (μM), nanomolar (nm), picomolar (pcM) or normal (n), all volumes are given in liters (l), milliliters ( ml) or microliters (μl) and linear measurements in millimeters (mm), unless otherwise specified.

 The following examples illustrate the isolation and synthesis of cryptophycin compounds, as well as their use as therapeutic agents in accordance with the invention.

In the extracts of about 1000 blue-green algae (cyanobacteria), which were tested for antitumor activity, it was found that the lipophilic extract of Nostoc sp. GSV 224 has the strongest cytotoxicity ³ , exhibiting minimum inhibitory concentrations (MICs) of 0.24 ng / ml against KB, against the human nasopharyngial carcinoma cell line, and 6 ng / ml against LoVo, the human calorectal adenocarcinoma cell line. More importantly, this extract exhibits significant selective tumor cytotoxicity in the Corbett assay. A bioanalysis recorded by reverse phase chromatography of an algal extract resulted in a fraction that predominantly represented Cryptophycin 1, a potent fungicide that was previously isolated from Nostoc sp. ATCC 53789 by Merck researchers and was found to be very active against strains of Cryptococcus.

Cryptophycin 1 was responsible for most of the cytotoxic activity of the crude algal extract of Nostoc sp. GSV 224 and pure compound, showing IC ₅₀ values of 3 and 5 pkg / ml against KB and LoVo, respectively. In a Corbett analysis, it was found that Cryptophycin 1 was highly selective for the tumor and equally cytotoxic against drug sensitivity and drug resistance of tumor cells. Immunofluorescence analysis shows that Cryptophycin 1 interacts with the cellular target in the same way as it does with vinblastine, but differs from the last drug in that it has a longer duration of action and does not form paracrystalline bodies. In preliminary in vivo experiments, Cryptophycin 1 exhibits very promising activity against tumors implanted in mice.

 Minimal amounts of some other cryptophycin compounds were present in Nostoc sp. GSV 224. Twenty-one of these compounds can be isolated in sufficient amounts to determine the structure and antitumor assessment in vitro by extracting algae with a 1: 5 mixture of dichloromethane / acetonitrile and reverse phase liquid chromatography-gel chromatography. Cryptophycins 2, 3, 4, 16, 17, 18, 19, 21, 23, 24, 26, 28, 29, 30, 31, 40, 43, 45, 49, 50 and 54, accompanying Cryptophycin 1, in the fraction, 65:35 acetonitrile / water eluted from a reverse phase flash column. Cryptophycins 2, 3, 4, 5, 6, and 7 were found only if the algae was extracted with methanol and reverse phase chromatography was performed in a methanol / water mixture. Cryptophycin 2, 3, 4, 5, and 6 were eluted with a 3: 1 methanol / water mixture, and Cryptophycin 7, found in an earlier, less toxic fraction, was eluted with a 1: 3 methanol / water mixture. Acyclic Cryptophycin 5, 6, and 7 were random products generated by the decomposition of Cryptophycin 1 during the isolation procedure.

Cryptophycin 3 and 5 were identical with fungicidal semisynthetic compounds prepared from Cryptophycin 1 by Merck researchers ^{8, 9} . Cryptophycin 3 was prepared by treating Cryptophycin 1 with a zinc-copper pair or diphosphoretetraiodide ⁸ , Cryptophycin 5 was prepared by methanolysis of Cryptophycin 1 ⁹ .

Example 1. The definition of the structure
Determination of the structures of new compounds, namely Cryptophycin 2, 4, 6, 7, 8, 9, 10, 12, 14, 16, 17, 18, 19, 21, 23, 24, 26, 28, 29, 30, 31, 40, 43, 45, 49, 50 and 54, as well as those that were previously disclosed, were carried out in a direct manner using a methodology that is well known to those skilled in the art. Mass spectral data were consistent with molecular compositions. The data of proton magnetic resonance and carbon nuclear magnetic resonance obtained from the COSY, HMQC, NMVC, and NOESY spectra make it possible to collect all the basic structures of these depsipeptide-type compounds. The presence of various hydroxyl and amino acid units in each compound was confirmed by gas chromatographic mass spectral analysis. Complete structures, including absolute stereochemistry, were determined using a combination of chemical destruction methods and spectral analytical methods of the corresponding derivatives of cryptophycin compounds.

Example 2. The dependence of the structure-activity (SAR)
To study the structural characteristics of Cryptophycin 1 necessary for optimal activity, all of the compounds described here were evaluated for cytotoxicity with respect to the KB cell line (human nasopharyngial carcinoma), LoVo (human colon carcinoma) and SKOV3 (ovarian human carcinoma). IC ₅₀ values are shown in Tables 1 and 2. Comparison of cytotoxicity shows that the intact (inactive) macrolide ring, epoxy and methyl groups and the double bond in 7,8-epoxy-5-hydroxy-6-methyl-8-phenyl-2- an octene acid unit (see unit A in Figure 1), chlorine and O-methyl groups in the 3- (3-chloro-4-methoxyphenyl) alanine unit (unit B), a methyl group in the 3-amino-2-methylpropionic acid unit ( unit C), and the isobutyl group in the leucine acid unit (unit D) of Cryptophycin 1, are necessary for optimal cytotoxicity. The strong cytotoxicity of Cryptophycin 8 is most likely due to chlorohydrin functionality, which acts as a masking agent for the epoxide.

The most active compounds were also evaluated for selective cytotoxicity against four different cell types, namely, murine leukemia (L1210 or P388), murine solid tumor (colon adenocarcinoma 38, duct 03 pancreatic adenocarcinoma, M16 / M17 breast adenocarcinoma), human solid carcinoma tumor (colon CX-1, HCT8, N116; earth fault P125 lung, breast MX-1, MCF-7) and fibroblast (LML) weak malignancy using ⁴ Corbett assay, diffuse disk assay modeled after analysis of chno used in antifungal and antibacterial testing. The results shown in Table 1 indicate that Cryptophycins 1-5 and 8 are not selective for either a solid tumor or leukemia, but are much more active against tumor cell lines, including those tumor cell lines that resist drug such as M17. None of the compounds shows an inhibition zone for any cell lines of a solid tumor, such a zone that would constitute a zone of 250 units, i.e. 7.5 mm, larger than the inhibition zone for the leukemia cell line. Cryptophycins 1-5 and 8, however, exhibit markedly large inhibition zones (zone of 400 units) for all tumor cell lines, compared with the inhibition zone for LML fibroblast.

 Diagnostically, it was found that LML behaves more like a normal cell than a tumor cell with respect to clinically useful cytotoxic agents (see Corbett assay data for 5-fluorouracil, etopside, and taxol in Table 1). Since various cytotoxicities for the zone were greater than 250 units, Cryptophycins 1-5 and 8 were tumor selective. Therefore, these compounds have become candidates for in vivo testing.

 Cryptophycin 1 is active against a wide range of murine and human tumors implanted in mice, including drug resistance tumors (Table 3). Cryptophycin 1 exhibits excellent activity against the five early stages of murine tumors, namely, cecum adenocarcinomas # 38 and # 51, a taxol-sensitive mammary tumor that resists Taxol # 16 / C / RP, and pancreatic double adenocarcinoma # 03 and two early stages of human tumors tested in mice with (SCID) severe combined immunodeficiency, namely MX-1 breast and H125 lung adenosarcoma, showing T / C tumor predisposition values (average tumor predisposition in the treated women otnyh / average tumor predisposition in untreated animals) that comprise less than 10%.

T / C values that are less than 42% are considered active according to NCl standards; T / C values that are less than 10% are considered to have excellent activity and potential clinical activity according to NCl standards ⁴ . Two of the three trials show a total Log kill (tumor cell) of 2.0. The total Log kill value is determined as T-C / 3.2 Td, where T represents the average time in days for tumors of the treated group to reach a tumor of 750 mg, C represents the average time in days for tumors of the control group to reach a tumor of 750 mg, and Td represents the doubling time of the tumor volume. The total values of Log destruction above 2.8, 2.0-2.8, 1.3-1.9, 0.5-0.8 and below 0.5 with a drug treatment duration of 5-20 days are considered ++++, +++, ++, + and - (inactive), respectively . A characterization of activity from +++ to ++++, which is an indication of clinical activity, is necessary for partial influence or complete regression of masses of 100-300 mg in size of most transplanted solid tumors of mice.

 Cryptophycin 8 is also active against a wide range of tumors transplanted into mice (Table 4). It exhibits excellent activity against all tumors tested to date, showing T / C tumor predisposition values of less than 10%, but more importantly, the total characteristics of Log kill activity from +++ to ++++ and, in some cases, cure.

 Good in vivo activity was also detected with Cryptophycin 35 in one trial to date.

 The lethal toxicity observed during the testing of Cryptophycin 1 and 8 was attributed to leukopenia, which is common for all clinically used antitumor drugs.

Example 3. The state of culture
Nostoc sp. GSV 224 was obtained from Professor CP Wolk, MSU-DOE Plant Research Laboratory, Michigan State University. Nostoc sp. ATCC 53789 was purchased from
American Type Culture Collection. A 1 L algae culture flask was used to inoculate an autoclaved 20 L glass container containing an inorganic medium designated BG-11 ³ , the pH of which was adjusted to 7.0 NaOH. The cultures were continuously irradiated with light with an incident light intensity of 200 μmol photons m ^-2 sec ^-1 (photosynthetically active radiation) from a bench of cold white fluorescent tubes and aerated at a speed of 5 l / min with a mixture of CO ₂ and air at a temperature of 24 ^o +/- 1 ^o C. Typically, the culture was collected by filtration after 21 days. Yields of lyophilized Nostoc sp. GSV 224 and ATCC 53789 averaged 0.61 and 0.3 g / l of culture, respectively.

Example 4. Isolation
Method A: Lyophilized Nostoc sp. GSV 224 was extracted with 2 L of a 1: 5 CH ₂ Cl ₂ / CH ₃ CN mixture for 48 hours and the extract was concentrated in vacuo to give a dark green solid. The residue (1 g; KB MIC 0.24 ng / ml) was placed in a silicon column coated with ODS (55 g, 7 • 5 cm) and flash chromatographed with 1: 3 CH ₃ CN / H ₂ O (0.8 L), 1: 1 CH ₃ CN / H ₂ O (0.8 L), 65:35 CH ₃ CN / H ₂ O (1.0 L), MeOH (0.8 L) and CH ₂ Cl ₂ (0.5 L). The fraction that was eluted 65:35 CH ₃ CN / H ₂ O (420 mg KB MIC 14 pg / ml) was subjected to reverse phase liquid chromatography (Econosil C18, 10 μm, 25 cm x 21, 5 mm, UV detection at 250 nm, 65:35 CH ₃ CN / H ₂ O, flow rate 6 ml / min) to obtain Cryptophycin 1 (t _R 49.3 min, 220 mg) and a number of impurity fractions. The fraction eluted from the Econosil C18 column at t _R 28.8 min was further purified by normal phase liquid gel permeation chromatography (Econosil silica gel 5 μm cartridge, 250 • 4.6 mm, 6: 4 ethyl acetate / hexane, 3 ml / min) to obtain Cryptophycin 16 (3.0 mg). The fraction eluted from the Econosil C18 column at t _R 32.5 min was subjected to liquid chromatography on an Econosil silica gel column using a 55:45 ethyl acetate / hexane mixture at 3 ml / min to give Cryptophycin 24 (0.8 mg) . The fraction eluted from the Econosil C18 column at t _R 35.5 min was twice subjected to liquid gel permeation chromatography on an Econosil silica gel column first using a 1: 1 ethyl acetate / hexane mixture at a speed of 3 ml / min and then using a mixture of 4 : 6 ethyl acetate / methylene chloride at a rate of 2.5 ml / min to give Cryptophycin 23 (1.2 mg) and Cryptophycin 43 (0.1 mg). The fraction eluted from the Econosil C18 column at t _R 39.5 min was subjected to liquid chromatography on an Econosil silica gel column using a 1: 1 ethyl acetate / hexane mixture at 3 ml / min to give Cryptophycin 2 (6 mg) and Cryptophycin 21 (14 mg) and a complex mixture of cryptophycin eluted at t _R 32.5 min. This last fraction, collected from 400 g of dry algae, was chromatographed on a selective semi-preparative column (partisil C18, 250 • 9.4 mm, 10 microns) with 65:35 water / acetonitrile and a reverse phase analytical column (Econosil, 150 • 4.6 mm, 5 μm) with a 5: 4: 1 mixture of water / acetonitrile / methanol at a rate of 1.3 ml / min to obtain Cryptophycin 50 (t _R 34.8, 0.4 mg) and Cryptophycin 40 (t _R 38.8 min, 0.3 mg). The fraction eluted from the Econosil C18 column at t _R 44.5 min was twice subjected to liquid gel permeation chromatography on an Econosil 1: 1 silica gel column with ethyl acetate / hexane at a rate of 3 ml / min to obtain Cryptophycin 17 (0.3 mg). Purification by liquid phase gel permeation chromatography with normal phase, the fraction eluted from the Econosil C18 column at t _R 54.5 min, as a shoulder to Cryptophycin 1, gave Cryptophycin 45 (t _R 6.7 min, 0.1 mg), Cryptophycin 26 (t _R 8.9 min, 0.5 mg) and Cryptophycin 54 (t _R 19.8 min, less than 0.1 mg) with elution with a 1: 1 ethyl acetate / hexane mixture. The fraction eluted from the Econosil C18 column as a wide peak (t _R 58 to 70 minutes) was analyzed by liquid gel permeation chromatography on an Econosil silica gel column with 43:57 ethyl acetate / hexane at 2.5 ml / min to give Cryptophycin 4 (t _R 19.6 min, 1.5 mg), Cryptophycin 31 (t _R 9.4 min, 0.8 mg), Cryptophycin 19 (t _R 25/8 min, 1.5 mg), Cryptophycin 40 (t _R 28 min , 0.1 mg), Cryptophycin 28 (t _R 29.0 min, 0.5 mg) and Cryptophycin 29 impurities (t _R 52.5 min, 2.0 mg) and Cryptophycin 30 (t _R 49 min, 3.0 mg). Pure Cryptophycins 29 and 30 were obtained after treatment by reverse-phase gel permeation chromatography (Econosil C18, 10 µ, 250 x 10 mm, 3: 1 methanol / water). The fraction eluted from the Econosil C18 column at t _R 78.9 min was subjected to gel permeation chromatography on an Econosil silica gel column to obtain Cryptophycin 3 (t _R 16.4 min, 3.0 mg). The fraction eluted from the Econosil C18 column at t _R 82.8 min was subjected to gel permeation chromatography on an Econosil silica gel column with 45:55 ethyl acetate / hexane at a rate of 3 ml / min and gave Cryptophycin 18 (t _R 19.2, 0.8 mg).

Method B:
Lyophilized Nostoc sp. GS V 224 (12.23 g) was twice extracted with 700 ml and 400 ml portions of MeOH for 12 and 5 hours (h), respectively. The extracts were combined and concentrated in vacuo to give 1.84 g of a dark green solid, which was partitioned between water and CH ₂ Cl ₂ . The lipophilic portion (0.65 g, KB MIC 0.24 ng / ml) was loaded into an ODS-coated silica gel column (55 g, 7 × 5 cm) and flash chromatographed with 1: 3 MeOH / H ₂ O, 1: 1 MeOH / H ₂ O (0.8 L), 3: 1 MeOH / H ₂ O (0.8 L), MeOH (0.8 L) and CH ₂ Cl ₂ (0.5 L). A fraction that was eluted with 3: 1 MeOH / H ₂ O (22 mg, KB MIC 14 pg / ml), which was considered to be essentially all having cytotoxic activity, was subjected to reverse phase gel chromatography ( Econosil C18, 10 microns, 250 • 10 mm, detection using UV at 250 nm, flow rate 3 ml / min) using 1: 5 MeOH / H ₂ O as eluent, gave Cryptophycin 7 (t _R 7.6 min, 0 , 2 mg), 5 (t _R 15.4 min, 2.3 mg), 2 (t _R 16.0 min, 1.0 mg), 1 (t _R 19.0 min, 12.0 mg), 4 (t _R 26.5 min, 1 , 2 mg) and 3 (t _R 30.2 min, 1.4 mg). The fraction (8.1 mg), which was eluted from the flash column using 1: 3 MeOH / H ₂ O, from one of the cultures, showed moderate cytotoxicity (KB MIC 2 μg / ml). Purification by liquid gel permeation chromatography using 2: 3 MeOH / H ₂ O as an eluent gave Cryptophycin G (7, t _R 6.0 min, 2.4 mg).

 Example 5. Spectral data for Cryptophycin 1-7.

 Letters in italics refer to links A-D (in figure 1).

 Cryptophycin 1.

[α] _D +33.8 ^° (MeOH, C 1.83); UV λ _max (ε) 208 (42,400), 218 (33,700), 228 (23,800), 280 (2,210); CD [θ] ₂₀₂ +15.900, [θ] ₂₀₆ +64.900, [θ] ₂₁₄ +26.900, [θ] ₂₂₄ +46.300, [θ] ₂₃₇ +10.500. IR (CHCl ₃ ) ν _max 3425, 2963, 1751, 1719, 1677, 1502, 1259 cm ^-1 . EIMS m / z (relative intensity) 654/656 (20/9), 412/414 (33/12), 280/282 (31/12), 227 (80), 195/197 (92/44), 91 (100); high resolution EIMS m / z 654.2665 (calculated for С ₃₅ Н ₄₃ СlN ₂ О ₈ , 4.3 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7.8-epoxy-5-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.74 (2, dt; 15.5 and 0.9), 6.68 (3, ddd; 15.5, 9.6 and 5.2), 2.45 (4, ddd; 14.2, 11.1 and 9.6), 2.55 (4, brdd; 14.2 and 5.2). 5.16 (5, ddd; 11.1, 4.9 and 1.9), 1.80 (6, m), 1.14 (6-Me, d; 7.1). 2.92 (7, dd; 7.5 and 2.0), 3.69 (8, d; 2.0), 7.25 (10/14, m), 7.34-7.39 (11/12/13, m); leucic acid (D) 4.83 (2, dd; 6.8 and 3.3), 1.70 (3, m), 1.36 (3, m), 1.70 (4, m), 0.86 (5, d; 6.6), 0.85 (4- Me, d; 6.6); 3-amino-2-methylpropionic acid (C) 2.71 (2, m), 1.22 (2-Me, d; 7.1), 3.30 (3, ddd; 13.4, 5.8 and 3.8), 3.48 (3, ddd; 13.4, 6.3 and 5.8), 6.93 (3-NH, brt; 5.8); 3-chloro-4-methoxyphenylalanine (B) 4.80 (2, ddd; 8.7, 7.3 and 5.4), 5.61 (2-NH, d; 8.7), 3.03 (3, dd; 14.4 and 7.3), 3.13 (3, dd ; 14.4. And 5.4.), 7.21 (5, d; 2.1), 3.87 (7-OCH ₃ , s), 6.83 (8, d; 8.5), 7.07 (9, dd; 8.5 and 2.1). ¹³ C NMR (CDCl ₃ ): δ unit (carbon position) A 165.3 (1), 125.3 (2), 141.0 (3), 36.7 (4), 76.2 (5), 40.6 (6), 13.5 (6-Me ), 63.0 (7), 59.0 (8), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.5 (12); D 170.7 (1), 71.3 (2), 39.4 (3), 24.5 (4), 22.9 (5), 21.3 (4-Me); C 175.6 (1), 38.2 (2), 14.1 (2-Me), 41.1 (3); B 170.9 (1), 53.6 (2), 35.0 (3), 129.7 (4), 131.0 (5), 122.4 (6), 154.0 (7), 56.1 (7-OCH ₃ ), 112.2 (8), 128.4 (9).

Cryptophycin 2
[α] _D +20.4 ^o (MeOH, s 0.54); UV λ _max (ε) 206 (43,800), 218 (37,500), 232 (22,900), 278 (2,410); CD [θ] ₂₀₃ +54,100, [θ] ₂₁₂ +16,500, [θ] ₂₂₅ +53,600, [θ] ₂₃₆ - 14,000. IR (CHCl ₃ ) ν _max 3423, 3029, 2961, 1742, 1724, 1678, 1512, 1258 cm ^-1 . EIMS m / z (relative intensity, assignment) 620 (11, M ⁺ ), 431 (3), 378 (8), 377 (6), 311 (11), 246 (10), 244 (8), 227 (14), 195 (17), 161 (84, CH ₃ O — C ₃ H ₄ —CH = CH = CO ⁺ ), 121 (79, CH ₃ O — C ₆ H ₄ —CH ₂ ⁺ ), 91 ( 100); high resolution EIMS m / z 620.3094 (calculated for C ₃₅ H ₄₄ N ₂ O ₈ , 0.3 mmu error); 161.0605 (calculated for C ₁₀ H ₉ O ₂ , - 0.2 mmu error); 121.0658 (calculated for C ₈ H ₉ O, - 0.4 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7,8-epoxy-5-hydroxy-6-methyl-8-phenyl-octenoic acid (A) 5.71 (2, dd; 15.4 and 1.3), 6.70 (3. ddd; 15.4, 10.2 and 5.0), 2.45 (4, m), 2.55 (4, m), 5.18 (5, ddd; 11.3, 4.8 and 2.0), 1.79 (6 , m), 1.14 (6-Me, d; 7.0), 2.92 (7, dd; 7.7 and 2.0), 3.68 (8, d; 2.0), 7.24 (10/14, m), 7.34-7.39 ( 11/12/13, m); leucic acid (D) 4.82 (2, dd; 10.1 and 3.7), 1.70 (3, m), 1.33 (3, m), 1.70 (4, m), 0.86 (5, d; 6.4), 0.84 ( 4-Me, d; 6.4); 3-amino-2-methylpropionic acid (C) 2.68 (2, m), 1.23 (2-Me, d; 7.3), 3.39 (3-H ₂ , m), 7.02 (3-NH, brt; 6.0); O-methyl tyrosine (B) 4.79 (2, ddd; 8.1, 7.0 and 5.7), 5.55 (2-NH, d; 8.1), 3.07 (3, dd; 14.5 and 7.0), 3.13 (3, dd; 14.5 and 5.7, 7.10 (5/9, d; 8.6), 6.81 (6/8, d; 8.6), 3.78 (7-OCH ₃ , s). ¹³ NMR (CDCl ₃ ): δ unit (carbon position) A 165.1 (1), 125.1 (2), 141.1 (3), 36.7 (4), 76.0 (5), 40.7 (6), 13.6 (6-Me), 63.0 (7), 59.0 (8), 136.7 (9 ), 125.6 (10/14), 128.7 (11/13), 128.5 (12); D 170.6 (1), 71.3 (2), 39.4 (3), 24.5 (4), 21.3 (5), 22.9 (4 -Me); C 176.0 (1), 38.1 (2), 14.2 (2-Me), 40.7 (3); B 171.1 (1), 53.3 (2), 35.3 (3), 131.0 (4), 130.2 ( 5/9), 114.1 (6/8), 158.6 (7), 55.2 (7-OCH ₃ ).

 Cryptophycin 3.

[α] _D +20.3 ^° (MeOH, s 1.13); UV λ _max (ε) 206 (51,700), 218 (31,200), 230 (22,900), 246 (18,800), 280 (3,230); CD [θ] ₂₀₅ +50,000, [θ] ₂₁₂ - 390, [θ] ₂₁₈ -47,200, [θ] ₂₃₃ - 100, [θ] ₂₅₁ +33,400, [θ] ₂₇₁ +4,310. IR (CHCl ₃ ) ν _max 3417, 2926, 1742, 1721, 1676, 1499, 1336 cm ^-1 . EIMS m / z (relative intensity) 638/640 (2 / 0.7, M ⁺ ), 412/414 (63/19), 280/282 (15/5), 227 (100), 195 (63), 91 (98); high resolution EIMS m / z 638.2764 (calculated for C ₃₅ H ₄₃ ClN ₂ O ₇ , - 0.5 mmu error), 412.1516 (calculated for C ₂₀ H ₂₇ ClNO ₆ , 1.1 mmu error), 227.1293 (calculated for C ₁₅ H ₁₇ NO , 1.0 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.77 (2, d; 15.5), 6.68 (3, ddd; 15.5, 9.5 and 5.3), 2.37 (4, m), 2.54 (4, m), 5.01 (5, ddd; 11.4, 6 and 1.5), 2.56 (6, m ), 1.14 (6-Me, d; 7.0), 6.01 (7, dd; 15.8 and 8.8), 6.41 (8, d; 15.8), 7.28-7.34 (10/11/13/14, m), 7.23 (12, m); leucine acid (D) 4.84 (2, dd; 10.1 and 3.6), 1.62 (3, m), 1.36 (3, m), 1.62 (4, m), 0.77 (5, d; 6.5), 0.73 (4- Me, d; 6.3); 3-amino-2-methylpropionic acid (C) 2.71 (2, m), 1.22 (2-Me, d; 7.3), 3.28 (3, dt; 13.5 and 7.0), 3.50 (3, ddd; 13.5, 4.9 and 4), 6.93 (3-NH, brt; 6.3); 3-chloro-4-methoxyphenylalanine (B) 4.82 (2, m), 5.64 (2-NH, d; 8.8), 3.05 (3, dd; 14.5 and 7.0), 3.13 (3, dd; 14.5 and 5.5), 7.22 (5, d; 2.2), 3.87 (7-OCH ₃ , s), 6.84 (8, d; 8.5), 7.08 (9, dd; 8.5 and 2.2). ¹³ C NMR (CDCl ₃ ): δ unit (carbon position) A 165.4 (1), 125.2 (2), 141.4 (3), 36.5 (4), 77.1 (5), 42.3 (6), 17.3 (6-Me ), 130.1 (7), 130.0 (8), 136.7 (9), 126.1 (10/14), 128.6 (11/13), 128.4 (12); D 170.1 (1), 71.6 (2), 39.5 (3), 24.5 (4), 21.2 (5), 22.7 (4-Me); C 175.6 (1), 38.3 (2), 14.0 (2-Me), 41.2 (3); B 170.9 (1), 53.5 (2), 35.1 (3), 129.8 (4), 131.0 (5), 122.4 (6), 154.0 (7), 56.1 (7-OCH ₃ ), 112.2 (8), 127.6 (9).

Cryptophycin 4
[α] _D +36.7 ^° (MeOH, s 1.93); UV λ _max (ε) 206 (41.800), 228 (25.000), 240 (21,200), 248 (22,500), 280 (3,000), 290 (1,230); CD [θ] ₂₀₅ +63,900, [θ] ₂₁₁ +3,040, [θ] ₂₁₈ -71,900, [θ] ₂₂₉ -11,700, [θ] ₂₃₄ -130, [θ] ₂₅₂ +47,500, [θ] ₂₇₀ , +5,400 . IR (CHCl ₃ ) ν _max 3410, 2962, 2917, 1741, 1718, 1678, 1511, 1251 cm ^-1 . EIMS m / z (relative intensity) 604 (2, M ⁺ ), 378 (74), 246 (11), 227 (46), 161 (100), 91 (96); high resolution EIMS m / z 604.3127 (calculated for C ₃₅ H ₄₄ N ₂ O ₇ , 2.2 mmu error), 378.1910 (calculated for C ₂₀ H ₂₈ NO ₆ , 0.7 mmu error), 227.1293 (calculated for C ₁₅ H ₁₇ NO, 1.7 mmu error), 161.0605 (calculated for С ₁₀ Н ₉ O ₂ , - 0.2 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.74 (2, dd; 15.3 and 1.2), 6.71 (3, ddd; 15.3, 10.3 and 5.0), 2.37 (4, m), 2.53 (4, m), 5.03 (5, ddd; 11.2, 6.4 and 2.0), 2.55 (6, m ), 1.13 (6-Me, d; 6.8), 6.01 (7, dd; 15.8 and 8.8), 6.40 (8, d; 15.8), 7.28-7.37 (10/11/13/14, m), 7.22 ( 12, m); leucine acid (D) 4.84 (2, dd; 10.1 and 3.6), 1.65 (3, m), 1.34 (3, m), 1.65 (4, m), 0.75 (5, d; 6.5), 0.72 (4- Me, d; 6.3); 3-amino-2-methylpropionic acid (C) 2.69 (2, m), 1.22 (2-Me, d; 7.5), 3.39 (3-H ₂ , m), 7.03 (3-NH, brt; 6.0); O-methyl tyrosine (B) 4.79 (2, m), 5.61 (2-NH, d; 7.8), 3.08 (3, dd; 14.5 and 7.0), 3.13 (3, dd; 14.5 and 5.3), 7.11 ( 5/9, d; 8.8), 6.81 (6/8, d; 8.8), 3.78 (7-OCH ₃ , s). ¹³ C NMR (CDCl ₃ ): δ unit (carbon position) A 165.3 (1), 125.1 (2), 141.5 (3), 36.5 (4), 77.1 (5), 42.3 (6), 17.3 (6-Me ), 130.1 (7), 131.8 (8), 136.7 (9), 126.2 (10/14), 128.7 (11/13), 127.6 (12); D 170.8 (1), 71.6 (2), 39.5 (3), 24.5 (4), 21.2 (5), 22.7 (4-Me); C 175.9 (1), 38.2 (2), 14.2 (2-Me), 40.9 (3); B 171.2 (1), 53.8 (2), 35.3 (3), 131.0 (4), 130.2 (5/9), 114.1 (6/8), 158.6 (7), 55.2 (7-OCH ₃ ).

Cryptophycin 5
[α] _D +36.0 ^o (MeOH, s 0.55); UV λ _max (ε) 206 (45.600), 218 (37.700), 280 (3.790), 286 (3.480), 325 (2.080); CD [θ] ₂₀₃ +7,710, [θ] ₂₀₆ +29,000, [θ] ₂₁₀ +21,400, [θ] ₂₂₂ +59,800, [θ] ₂₃₄ +12,800, [θ] ₂₄₁ +13,700. IR (CHCl ₃ ) ν _{max /} 3426, 2958, 1728, 1672, 1502, 1259 cm ^-1 . EIMS m / z (relative intensity) 686/688 (0.1510.05), 655/657 (1 / 0.3), 654/656 (1.5 / 0.5), 311/313 (75/27), 195 (66), 155 (54), 121 (51), 91 (100); high resolution EIMS m / z 686.2983 (calculated for C ₃₆ H ₄₇ ClN ₂ O ₉ , - 1.3 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz 7.8-epoxy-5-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.87 (2 , d; 15.3), 6.72 (3, dt; 15.3 and 6.8), 2.60 (4, m), 2.52 (4, ddd; 15.2, 7.8, and 6.8), 5.11 (5, ddd; 12.3, 7.8, and 7.1 ), 1.87 (6, m), 1.12 (6-Me, d; 7.1), 2.91 (7, dd; 7.3 and 2.1), 3.70 (8, d; 2.1), 7.24 (10/14, brd; 7.4) , 7.29-7.36 (11/12/13, m); leucinic acid (D) 4.09 (2, m), 2.86 (2-OH, brd, 6.1), 1.83 (3, m), 1.42 ( 3, m), 1.86 (4, m), 0.90 (5, d; 6.6), 0.87 (4-Me, d; 6.8); 3-amino-2-methylpropionic acid (C) 3.64 (1-OCH ₃ , s), 2.60 (2, m), 1.07 (2-Me, d; 7.3), 3.27 (3, ddd; 13.5, 8.0 and 5.5), 3.39 (3, m), 6.32 (3-NH, t; 5.4 ); 3-chloro-4-methoxyphenylalanine (B) 4.59 (2, dt; 6 and 7.5), 6.30 (2-NH, d; 7.5), 2.95 (3, dd; 13.6 and 7.5), 3.0 (3, dd; 13.6 and 6.0), 7.2 (5, d; 2.1), 3.86 (7-OCH ₃ , s), 6.84 (8, d; 8.5), 7.05 (9, dd, 8.5; 2.1). ¹³ C NMR (CDCl ₃ ): δ unit (carbon position) A 164.8 (1), 126.5 (2), 139.2 (3), 34.4 (4), 75.5 (5), 39.2 (6), 12.9 (6-Me ), 63.3 (7), 58.7 (8), 136.8 (9), 125.7 (10/14), 128.6 (11/13), 128.4 (12); D 175.1 (1), 69.2 (2), 43.2 (3), 24.3 (4), 21.2 (5), 23.2 (4-Me); C 175.4 (1), 51.9 (1-OMe), 39.1 (2), 14.7 (2-Me), 41.6 (3); at 170.6 (1), 54.6 (2), 37.4 (3), 129.5 (4), 131.0 (5), 122.4 (6), 154.1 (7), 56.1 (7-OMe), 112.2 (8), 128.4 (9).

Cryptophycin 6
[α] _D +17.1 ^° (MeOH, C 1.1); UV λ _max (ε) 206 (40,000), 218 (30,100), 228 (21,400), 282 (2,430); CD [θ] ₂₀₃ +37,700, [θ] ₂₁₀ -5,430, [θ] ₂₁₃ -1,260, [θ] ₂₂₁ +24,100, [θ] ₂₃₂ +8,480, [θ] ₂₄₀ +13,400, [θ] ₂₅₄ +790. IR (CHCl ₃ ) ν _max 3425, 3006, 2956, 1726, 1672, 1641, 1502, 1462, 1259 cm ^-1 . MC with fast atom bombardment (thioglycerol) m / z, (relative intensity) 573/575 (13/6) [MH ₂ O] ⁺ , 217 (26), 91 (100). ¹ H NMR (CDCl ₃ ): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 5,7,8-trihydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.92 (2, dt; 15.0 and 1.5),
6.94 (3, dt, 15 and 7.5), 2.51 (4, m), 2.64 (4, m), 3.97 (5, ddd; 9.3, 6.5 and 4.5), 2.03 (6, m), 1.10 (6-Me, d; 6.5), 3.70 (7, dd; 9.0 and 7.5), 4.64 (8, d; 7.5), 7.33-7.39 (10/11/13/14, m), 7.28 (12, tt ; 6.5 and 2.0); 3-chloro-4-methoxyphenylalanine (B) 4.60 (2, td; 8.0 and 6.0), 6.09 (2-NH, brd; 8.0), 2.96 (3, dd; 13.8 and 8.0), 3.02 (3, dd; 13.8 and 6.0), 7.22 (5, d; 2.0), 3.86 (7-OCH ₃ , s), 6.84 (8, d; 8.5), 7.07 (9, dd; 8.5 and 2.0) 3-amino-2-methylpropionic acid (C) 3.63 (1-OCH ₃ , s), 2.58 (2, m), 1.07 (2-Me, d; 7.0), 3.24 (3, ddd; 13.8, and 6.5), 3.41 (3, ddd; 13.8 6.5 and 4.8), 6.21 (3-NH, brt; 6.5). ¹³ C NMR (CDCl ₃ ): δ unit (carbon position) A 165.2 (1), 125.6 (2), 141.3 (3), 36.9 (4), 82.5 (5), 46.3 (6), 14.3 (6-Me ), 85.1 (7), 84.8 (8), 140.9 (9), 125.8 (10/14), 128.6 (11/13), 127.8 (12); B 170.6 (1), 54.5 (2), 37.3 (3), 129.6 (4), 131.0 (5), 122.5 (6), 154.1 (7), 56.1 (7-OCH ₃ ), 112.2 (8), 128.5 (9) C 52.0 (1-OCH ₃ ), 175.4 (1), 39.2 (2), 14.7 (2-Me), 41.6 (3).

Cryptophycin 7
[α] _D -51.9 ^o (MeOH, s 0.89); UV λ _max (ε) 206 (23,400), 220 (14,900), 282 (1,670); CD [θ] ₂₀₂ +35,400, [θ] ₂₀₆ -1,730, [θ] ₂₁₁ -19,200, [θ] ₂₂₀ -15,800, [θ] ₂₃₂ +29,000, [θ] ₂₆₃ +2,040. IR (CHCl ₃ ) ν _max 3426, 2946, 1732, 1675, 1501, 1258 cm ^-1 . EIMS m / z (relative intensity) 455/457 (1 / 0.3, [M-2H ₂ O] ⁺ ), 105 (100), 77 (98); MS with fast atomic bombardment m / z (matrix tests) 496/498 [MH ₂ O + Na] ⁺ , (thioglycerol matrix) 474/476 [M-H ₂ O + H] ^{+ 1} H NMR (CO ₃ OD ): amino or hydroxy-acid unit α (carbon position, multiplicity; J in Hz) 5,7,8-trihydroxy-6-methyl-8-phenyl-2-octenoic acid) (A) 6.06 (2, ddd; 15.5, 1.3 and 1.0), 6.80 (3, dt; 15.5 and 7.5), 2.49 (4, m), 2.59 (4, m), 3.92 (5, ddd; 9.5, 6.3 and 4.7), 1.95 (6, m) , 1.08 (6-Me, d; 6.7), 3.59 (7, dd; 9.0 and 7.8), 4.56 (8, d; 7.8), 7.37 (10/14, brd; 7.3), 7.31 (11/13, brt ; 7.3), 7.24 (12, tt; 7.3 and 1.5); 3-chloro-4-methoxyphenylalanine (B) 4.52 (2, dd; 6.9 and 5.0), 2.93 (3, dd; 13.8 and 6.9), 3.15 (3, dd; 13.8 and 5.0), 7.20 (5, d; 2.2 ), 3.78 (7-OCH ₃ , s), 6.88 (8, d; 8.4), 7.08 (9, dd; 8.4 and 2.2). ¹³ C NMR (CO ₃ OD): δ unit (carbon position) A 167.4 (1), 127.6 (2), 140.9 (3), 37.9 (4), 84.0 (5), 47.6 (6), 14.4 (6- Me), 86.0 (7), 85.8 (8), 142.9 (9), 127.1 (10/14), 129.3 (11/13), 128.5 (12); B 177.6 (1), 57.3 (2), 38.2 (3), 132.8 (4), 132.1 (5), 122.9 (6), 155.0 (7), 56.5 (7-OCH ₃ ), 113.2 (8), 130.1 (9).

Cryptophycin 16
[α] _D -41.3 ^o (MeOH, s 5.2); UV λ _max (ε) 242 (4963), 280 (2430), 286 (2212), IR (pure substance) ν _max 3402, 3270, 2960, 1748, 1724, 1676, 1514, 1466, 1343, 1239, 1177 cm ^-1 ; EIMS m / z (relative intensity) 640/642 (66/27), 398/400 (47/16), 265 (55), 227 (93), 181 (100); high resolution EIMS m / z 640.25676 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₈ , -1.6 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7.8-epoxy-6-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.74 ( 2, d; 16), 6.67 (3, add; 15.3, 9.7 and 5.5), 2.45 (4, dt; 14.3 and 10.4), 2.55 (4, brdd; 14.3 and 5.3), 5.15 (5, ddd; 11.2, 4.8 and 1.8), 1.8 (6, m), 1.14 (6-Me, d; 7.0), 2.92 (7, dd; 7.5 and 2.0), 3.69 (8, d; 2.0), 7.24-7.26 (10/14 , m), 7.33-7.39 (11/12/13, m); 3-chloro-4-hydroxyphenylalanine (B) 4.8 (2, m), 5.64 (2-NH, d; 8.8), 3.03 (3, dd; 14.5 and 7.0), 3.11 (3, dd; 14.4 and 5.6), 7.17 (5, d; 2.2), 5.61 (7-OH, s), 6.91 (8, d; 8.3), 7.0 (9, dd; 8.3 and 2.2); 3-amino-2-methylpropionic acid (C) 2.71 (2, m), 1.22 (2-Me, d; 7.3), 3.28 (3, dt; 13.6 and 6.8), 3.49 (3, ddd; 13.6, 5 and 4.1), 6.92 (3-NH, br t; 6.1); leucine acid (D) 4.83 (2, dd; 10.1 and 3.3), 1.36 (3, m), 1.67-1.75 (3, m), 1.67-1.75 (4, m), 0.85 (5, d; 7.5 ), 0.86 (4-Me, d; 6.8). ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.3 (1), 125.3 (2), 141.0 (3), 36.7 (4), 76.2 (5), 40.6 (6), 13.5 (6-Me) , 63.0 (7), 59.0 (8), 136.8 (9), 125.6 (10/14), 128.7 (11/13), 128.6 (12); B 170.9 (1), 53.6 (2), 35.1 (3), 129.9 (4), 129.6 (5), 120.0 (6), 150.4 (7), 116.4 (8), 129.2 (9); S 175.6 (1), 38.3 (2), 14.1 (2-Me), 41.1 (3), D 170.8 (1), 71.3 (2), 39.4 (3), 24.6 (4), 21.3 (5), 22.9 (4-Me).

Cryptophycin 17
[α] _D -27.8 ^° (CHCl ₃ p. 0.37); UV λ _max (ε) 248 (14740), 268 (8100), 278 (3400), 284 (2840); IR (pure substance) ν _max 3412, 2958, 1750, 1723, 1668, 1504, 1463, 1290, 1177, 751 cm ^-1 ; EIMS m / z (relative intensity) 624/626 (10/3), 398/400 (95/35), 284 (100), 149 (95); high resolution EIMS m / z 624.26161 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₇ , -1.4 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.77 (2, d; 15.4), 6.67 (3, ddd; 15.4, 9.5 and 5.3), 2.37 (4, m), 4.99 (5, ddd; 11.2, 6.3 and 1.6), 2.54 (6, m), 1.14 (6-Me , d; 6.7), 6.01 (7, dd; 15.7, and 8.7), 6.41 (8, d; 15.9), 7.28-7.34 (10/11/13/14, m), 7.23 (12, m); 3-chloro-4-hydroxyphenylalanine (B) 4.82 (2, m), 5.63 (2-NH, d; 8.7), 3.12 (3, dd; 14.7 and 5.6), 3.03 (3 ', dd; 14.7, and 7.1 ), 7.18 (5, d; 2.0), 5.47 (7-OH, br s), 6.91 (8, d; 8.3), 7.02 (9, dd; 8.3 and 2.0); 3-amino-2-methylpropionic acid (C) 2.71 (2, m), 1.21 (2-Me, d '6.9), 3.25 (3, m), 3.52 (3', m), 6.89 (3-NH, br t; 6.1); leucic acid (D) 4.84 (2, dd; 9.6 and 3.1), 1.62 (3, m), 1.36 (3 ', m), 1.62 (4, m), 0.77 (5, d' 6.5), 0.73 (4-Me, d; 6.5); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.4 (1), 125.3 (2), 141.3 (3), 36.5 (4), 77.1 (5), 42.3 (6), 17.3 (6-Me) 130.0 (7), 129.9 (8), 136.7 (9), 126.2 (10/14), 128.6 (11/13), 127.6 (12); B 170.9 (1), 53.5 (2), 35.1 (3), 129.6 (4), 131.9 (5), 126.2 (6), 150.3 (7), 116.3 (8), 127.6 (9); C 175.9 (1), 38.4 (2), 13.9 (2-Me), 41.3 (3); D 170.9 (1), 71.6 (2), 39.5 (3), 24.5 (4), 21.2 (5), 22.7 (4-Me).

Cryptophycin 18
[α] _D +54.9 ^° (MeOH, s 0.93); UV λ _max (ε) 250 (20518), 284 (3857); IR (pure substance) ν _max 3411, 3271, 2966, 1746, 1728, 1668, 1505, 1463, 1258, 1178 cm ^-1 ; EIMS m / z (relative intensity) 638/640 (4.5 / 1.1), 412/414 (59/19), 280 (17), 227 (100); high resolution EIMS m / z 638.272934 (calculated for C ₃₅ H ₄₃ ClN ₂ O ₇ , 2.9 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.76 (2, d; 15.5), 6.65 (3, ddd; 15.4, 9.2 and 6.2), 2.38-2.47 (4, m), 5.08 (5, ddd; 10.6, 4.9 and 2.2), 2.58 (6, m), 1.15 (6-Me , d; 6.8), 6.07 (7, dd; 15.9 and 8.5), 6.43 (8, d; 15.9), 7.21-7.35 (10/11/12/13/14, m); 3-chloro-4-methoxyphenylalanine (B) 4.83 (2, m), 3.05 (3, dd; 14.5 and 7.1), 5.65 (2-NH, d; 8.7), 3.14 (3, dd; 14.4 and 5.5), 7.21 (5, d; 2.4), 3.86 (7-OCH ₃ , s), 6.83 (8, d; 8.3), 7.08 (9, dd; 8.3 and 2.2); 3-amino-2-methylpropionic acid (C) 2.73 (2, m), 1.23 (2-Me, d; 7.2), 3.23 (3, dt; 13.5 and 6.8), 3.56 (3, ddd; 13.5, 5.7 and 4.0), 6.85 (3-NH, dd; 7.1 and 6.2); leucine acid (D) 4.8 (2, d; 4.6), 1.86-1.89 (3, m), 0.94 (3-Me, d; 7.0), 1.20-1.26 (4, m), 1.39-1.44 (4, m ), 0.77 (5, d; 7.4), ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.5 (1), 125.2 (2), 141.5 (3), 36.4 (4), 77.7 (5 ), 41.9 (6), 17.1 (6-Me), 129.8 (7), 131.9 (8), 136.8 (9), 128.6 (10/14), 126.2 (11/13), 127.6 (12); B 170.0 (1), 53.5 (2), 35.1 (3), 129.9 (4), 131.1 (5), 122.4 (6), 153.9 (7), 56.1 (7-OCH ₃ ), 112.2 (8) , 128.5 (9); C 175.3 (1), 38.6 (2), 14.0 (2-Me), 41.4 (3); D 169.5 (1), 76.6 (2), 36.2 (3), 15.5 (3-Me), 24.2 (4), 14.0 (5).

Cryptophycin 19
[α] _D +62.6 ^o (MeOH, s 0.67); UV (MeOH) λ _max (ε) 204 (44900), 230 (17000), 248 (15600), 280 (2500); IR (pure substance) ν _max 3413, 3272, 2966, 1745, 1726, 1672, 1504, 1258, 1199, 1178, 1066, 692 cm ^-1 ; EIMS m / z (relative intensity) 624/626 (3.0 / 1.4), 398/400 (58/21), 280/282 (15/5), 227 (100), 195/197 (57/22); high resolution EIMS m / z 624.2585 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₇ , -1.8 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.76 (2, d; 15.2), 6.64 (3, ddd; 15.4, 9.1 and 6.2), 2.38 (4, m), 2.47 (4, m), 5.04 (5, ddd; 7.1, 5.1 and 1.8), 2.57 (6, m), 1.15 (6-Me, d; 6.9), 6.05 (7, dd; 15.8 and 8.5), 6.43 (8, d; 15.8), 7.29-7.35 (10/11/13/14, m), 7.23 (12, m); 3-chloro-4-methoxyphenylalanine (B) 4.84 (2, m), 5.67 (2-NH, d; 8.9), 3.04 (3, dd; 14.3 and 7.1), 3.14 (3, dd; 14.3 and 5.3), 7.22 (5, d; 2.0), 3.86 (7-OCH ₃ , s), 6.83 (8, d; 8.2), 7.08 (9, dd; 8.2 and 2.0); 3-amino-2-methylpropionic acid (C) 2.75 (2, m), 1.23 (2-Me, d; 7.1), 3.19 (3, m), 3.59 (3, m), 6.80 (3-NH, brt ; 6.7); 2-hydroxyisovalerianic acid (D) 4.73 (2, d; 4.2), 2.09 (3, m), 0.84 (4, d; 6.9), 0.95 (3-Me, d; 6.9). ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.5 (1), 125.3 (2), 141.3 (3), 36.3 (4), 77.7 (5), 42.0 (6), 17.1 (6-Me) 129.9 (7), 131.9 (8), 136.8 (9), 126.1 (10/14), 128.6 (11/13), 127.6 (12); B 171.0 (1), 53.4 (2), 35.1 (3), 130.0 (4), 131.1 (5), 122.4 (6), 153.9 (7), 56.1 (7-OMe), 112.2 (8), 128, 5 (9); C 175.1 (1), 38.7 (2), 13.9 (2-Me), 41.5 (3); D 169.6 (1), 76.9 (2), 29.8 (3), 19.0 (4), 16.7 (3-Me).

Cryptophycin 21
[α] +40.2 ^° (CHCl ₃ with 0.72); UV (MeOH) λ _max (ε) 240 (6700), 280 (2400), 288 (2100); IR (pure substance) ν _max 3403, 3279, 2957, 1731, 1673, 1503, 1464, 1409, 1372, 1258, 1174, 1065, 1023, 889 cm ^-1 ; EIMS m / z (relative intensity) 640/642 (10/4), 612 (5), 478 (15), 398 (40), 266 (33), 227 (76), 195 (95), 155 ( 100), 127 (90); high resolution EIMS m / z 640.2550 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₈ , -0.2 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7,8-epoxy-5-hydroxy-6-methyl-8-phenylloctanoic acid (A) 5.73 (2, d; 15.4), 6.68 (3, ddd; 15.0, 9.9 and 4.9), 2.45 (4, m), 2.56 (4, m), 5.19 (5, ddd; 11.2, 5.1 and 1.5), 1.80 (6, m), 1.14 (6-Me, d; 7.1), 2.92 (7, dd; 7.5 and 2.0), 3.68 (8, d; 1.8), 7.25 (10/14, m), 7.33-7.38 (11/12/13, m); 3-chloro-4-methoxyphenylalanine (B) 4.74 (2, ddd; 8.2, 6.8 and 6.2), 5.68 (2-NH, d; 8.6), 2.98 (3, dd; 14.3 and 7.7), 3.14 (3, dd ; 14.3, and 5.6), 7.21 (5, d; 2.0), 3.86 (7-OMe, s), 6.83 (8, d; 8.4), 7.07 (9, dd; 8.4 and 2.0); 3-aminopropionic acid (C) 2.56 (2, m), 3.51 (3, m), 3.45 (3, m), 6.90 (3-NH, br t; 5.8); leucine acid (D) 4.89 (2, dd; 10.0 and 3.3), 1.67 (3, m), 1.31 (3, m), 1.67 (4, m), 0.84 (5, d; 6.4), 0.83 ( 4-Me, d; 6.4); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.5 (1), 125.3 (2), 141.0 (3), 36.7 (4), 75.9 (5), 40.6 (6), 13.5 (6-Me) , 63.0 (7), 59.0 (8), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.5 (12); B 170.7 (1), 53.9 (2), 35.0 (3), 129.8 (4), 130.9 (5), 122.4 (6), 153.9 (7), 56.1 (7-OMe), 112.2 (8), 128.3 (9); C 172.6 (1), 32.4 (2), 34.4 (3); D 170.5 (1), 71.2 (2), 39.5 (3), 24.4 (4), 22.8 (5), 21.2 (4-Me).

Cryptophycin 23
[α] _D +47 ^° (MeOH, s 1.55); UV λ _max (ε) 240 (4571), 282 (2174), 290 (2177); IR (pure substance) ν _max 3284, 2960, 1747, 1724, 1653, 1540, 1490, 1339, 1272, 1174 cm ^-1 ; EIMS m / z (relative intensity) 674/675/678 (47/35/8), 432/434/436 (11/5/2), 299/301/303 (39/30/7), 227 ( 64), 215/217/219 (31/20/8), 141 (100); high resolution EIMS m / z 674.21643 (calculated for C ₃₄ H ₄ Cl ₂ N ₂ O ₈ , -0.3 mmu error). ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7,8-epoxy-5-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.77 ( 2, d; 15.4), 6.65 (3, ddd; 15.4, 9.3 and 6.0), 2.47 (4, dt; 14.2 and 10.2), 2.55 (4, br dd; 14.2 and 5.6), 5.13 (5, ddd; 11.0 , 4.6 and 1.6), 1.81 (6, m), 1.15 (6-Me, d; 6.9), 2.93 (7, dd; 7.6 and 2.0), 3.7 (8, d; 2.0), 7.22-7.26 (10 / 14, m), 7.32-7.39 (11/12/13, m); 3,5-dichloro-4-hydroxyphenylalanine (B) 4.81 (2, m), 5.69 (2-NH, d; 8.6), 3.11 (3, dd; 14.5 and 5.6), 3.50 (3, dd; 14.3 and 7.0 ), 7.13 (5/9, s), 5.78 (7-OH, s), 3-amino-2-methylpropionic acid (C) 2.73 (2, m), 1.22 (2-Me, d; 7.1), 3.19 (3, dt; 13.4 and 6.9), 3.58 (3, ddd; 13.6, 5.8 and 4.1), 6.82 (3-NH, br t; 5.9); leucine acid (D) 4.84 (2, dd; 9.9 and 3.2), 1.38 (3, m), 1.68-1.75 (3, m), 1.68-1.75 (4, m), 0.86 (4-Me, d; 6.7 ), 0.87 (5, d; 6.7). ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.4 (1), 125.4 (2), 140.9 (3), 36.7 (4), 76.3 (5), 40.6 (6), 13.5 (6- Me), 63.0 (7), 58.9 (8), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.6 (12); B 170.7 (1), 53.3 (2), 35.0 (3), 130.3 (4), 129.0 (5/9), 121.0 (6/8), 146.7 (7); C 175.3 (1), 38.4 (2), 13.9 (2-Me), 41.5 (3); D 170.8 (1), 71.3 (2), 39.4 (3), 24.6 (4), 21.3 (4-Me), 22.9 (5).

(положение углерода) A 165.4 (1), 125.3 (2), 141.0 (3), 36.7 (4), 75.9 (5), 40.6 (6), 13.4 (6-Me), 63.0 (7), 59.0 (8), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.5 (12); В 170.7 или 170.6 (1), 54.1 (2), 35.2 (3), 128.5 (4), 130.2 (5/9), 114.1 (6/8), 158.6 (7), 55.2 (7-ОМе); С 172.8 (1), 32.5 (2), 34.2 (3); D 170.6 или 170.7 (1), 71.2 (2), 39.5 (3), 24.4 (4), 21.3 (5), 22.8 (4-Me).Cryptophycin 24
[α] _D +48.8 ^° (CHCl ₃ , s 0.63); UV λ _max (ε) 228 (19006), 242 (8249), 274 (2351); IR (pure substance) ν _max 3400, 3284, 2959, 1732, 1678, 1652, 1514, 1248, 1178 cm ^-1 ; EIMS m / z (relative intensity, assignment) 606 (2, M ⁺ ), 364 (7), 161 (55, CH ₃ O-C ₆ H ₄ -CH = CH = CO ⁺ ), 121 (100, CH ₃ OC ₆ H ₄ -CH ₂ ⁺ ), 91 (68); high resolution EIMS m / z 606.2954 (calculated for C ₃₄ H ₄₂ N ₂ O ₈ , -1.3 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7,8-epoxy-5-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.70 ( 2, dd; 15.2 and 1.3), 6.70 (3, ddd; 15.2, 10.3 and 4.7), 2.43 (4, dt; 14.3 and 10.9), 2.56 (4, m), 5.20 (5, ddd; 11.3, 5.1 and 2.0), 1.79 (6, m), 1.14 (6-Me, d; 7.0), 2.92 (7, dd; 7.5 and 2.0), 3.68 (8, d; 2.0), 7.23-7.38 (10/11/12 / 13/14, m); O-methyl tyrosine (B) 4.73 (2, m), 5.58 (2-NH, d; 8.3), 3.03 (3, dd; 14.5 and 7.5), 3.14 (3, dd; 14.5 and 5.7), 7.11 (5 / 9, d; 8.6), 6.81 (6/8, d; 8.6), 3.78 (7-OMe, s); 3-aminopropionic acid (C) 2.55 (2-H ₂ , m), 3.42 (3, m), 3.53 (3, m) 6.97 (3-NH, br t; 5.7); leucine acid (D) 4.89 (2, dd; 9.9 and 3.5), 1.29 (3, m), 1.62-1.70 (3/4, m), 0.83 (5, d; 5.9), 0.84 (4-Me, d ; 6.1); ¹³ C NMR (CDCl ₃ ) unit

(carbon position) A 165.4 (1), 125.3 (2), 141.0 (3), 36.7 (4), 75.9 (5), 40.6 (6), 13.4 (6-Me), 63.0 (7), 59.0 (8 ), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.5 (12); B 170.7 or 170.6 (1), 54.1 (2), 35.2 (3), 128.5 (4), 130.2 (5/9), 114.1 (6/8), 158.6 (7), 55.2 (7-OMe); C 172.8 (1), 32.5 (2), 34.2 (3); D 170.6 or 170.7 (1), 71.2 (2), 39.5 (3), 24.4 (4), 21.3 (5), 22.8 (4-Me).

Cryptophycin 26
[α] _D +28.2 ^° (CHCl ₃ , s 1.31); UV λ _max (ε) 254 (14615), 284 (2949), IR (pure substance) ν _max 3299, 2960, 1732, 1644, 1504, 1258, 1209 cm ^-1 ; EIMS m / z (relative intensity) 656/658 (0.5 / 0.1, M ⁺ ), 638/640 (1.7 / 1.0), 525/527 (3.7 / 1.8), 412/414 (10/4), 280 / 282 (12/11), 227 (20), 195 (48), 131 (68); high resolution EIMS m / z 656.2836 (calculated for C ₃₅ H ₄₅ ClN ₂ O ₈ , 2.8 mmu error); 638.2712 (calculated for C ₃₅ H ₄₃ ClN ₂ O ₇ , 4.7 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 3,5-dihydroxy-6-methyl-8-phenyl-7-octenoic acid (A) 2.46 (2, dd; 14.8 and 7.8), 2.58 (2, dd; 14.8 and 3.0), 5.46 (3, m), 1.86-1.90 (4-H ₂ , m), 3.61 (5, m), 2.37 (6, m) , 1.14 (6-Me, d; 6.8), 6.06 (7, dd; 16 and 8.7), 6.47 (8, d; 16), 7.37 (10/14, br d; 7.9), 7.32 (11/13, br t; 7.6), 77.22-7.28 (12, m); 3-chloro-4-methyloxyphenylalanine (B) 4.73 (2, br dt; 6.4 and 8.1), 6.14 (2-NH, d; 8.6), 2.84 (3, dd; 14.4 and 8), 3.18 (3, dd; 14.4 and 6.3), 7.21 (5, d; 2.2), 3.85 (7-OMe, s), 6.82 (8, d; 8.6), 7.08 (9, dd; 8.6 and 2.2); 3-amino-2-methylpropionic acid (C) 2.87 (2, m), 1.19 (2-Me, d; 7.0), 3.01 (3, ddd; 13.4, 10.6 and 4.9), 3.73 (3, ddd; 13.4, 8.2 and 4.7), 6.72 (3-NH, br dd; 7.3 and 5.2); leucine acid (D) 4.95 (2, dd; 9.7 and 4.2), 1.62-1.72 (3, m), 1.79-1.84 (3, m), 1.62-1.72 (4, m), 0.90 (4-Me, d ; 6.4), 0.95 (5, d; 6.4). ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 170.0 (1), 41.5 (2), 71.4 (3), 37.3 (4), 71.9 or 71.8 (5), 43.6 (6), 16.6 (6- Me), 130.8 (7), 132.5 (8), 136.8 (9), 126.2 (10/14), 128.6 (11/13), 127.6 (12); B 170.9 (1), 53.2 (2), 34.7 (3), 130.3 (4), 131.1 (5), 122.2 (6), 153.8 (7), 56.1 (7-OMe), 112.2 (8), 128.5 (9); C 174.3 (1), 40.1 (2), 14.4 (2-Me), 42.5 (3); D 170.7 (1), 71.8 or 71.9 (2), 38.9 (3), 24.6 (4), 21.6 (4-Me), 22.9 (5).

Cryptophycin 28
[α] _D +65.6 ^° (MeOH, s 0.93); UV (MeOH) λ _max (ε) 204 (48000), 230 (19300), 248 (18700), 280 (3400); IR (pure substance) ν _max 3413, 3270, 2958, 1745, 1726, 1665, 1504, 1258, 1197, 1175, 1066, 694 cm ^-1 ; EIMS m / z (relative intensity) 624/626 (3.0 / 1.3), 412/414 (70/24), 280/282 (13/6), 213 (100), 195/197 (86/40); high resolution EIMS m / z 624.2626 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₇ , - 2.4 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-8-phenyl-2,7-octadiene acid (A) 5.78 (2, d; 15.6), 6.71 (3, ddd; 15.6, 9.9 and 5.4), 2.40 (4, m), 2.53 (4, m), 5.17 (5, m), 2.53 (6-H ₂ , br t; 6.7), 6.07 (7, dt; 15.8 and 7.4), 6.44 (8, d; 15.8), 7.27-7.38 (10/11/13/14, m) 7.22 (12, m); 3-chloro-4-methoxyphenylalanine (B) 4.82 (2, m), 5.72 (2-NH, d; 8.5), 3.04 (3, dd; 14.5 and 7.2), 3.14 (3, dd; 14.5 and 5.4), 7.22 (5, d; 2.0), 3.87 (7-OMe, s), 6.84 (8, d; 8.5), 7.08 (9, dd; 8.5 and 2.0); 3-amino-2-methylpropionic acid (C) 2.72 (2, m), 1.21 (2-Me, d; 7.2), 3.29 (3, dt; 13.5 and 7.0), 3.49 (3, ddd; 13.5, 4.9 and 3.8), 6.97 (3-NH, br t; 5.6); leucine acid (D) 4.82 (2, m), 1.40 (3, m), 1.62 (3, m), 1.62 (4, m), 0.76 (4-Me, d; 6.3), 0.74 (5, d; 6.3); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.4 (1), 125.2 (2), 141.2 (3), 38.5 (4), 73.5 (5), 38.6 (6), 124.1 (7), 133.8 (8), 136.7 (9), 126.1 (10/14), 128.6 (11/13), 127.6 (12); B 170.9 (1), 53.6 (2), 35.1 (3), 129.8 (4), 131.0 (5), 122.4 (6), 154.0 (7), 56.1 (7-OMe), 112.3 (8), 128.4 ( 9); C 175.6 (1), 38.3 (2), 14.0 (2-Me), 41.2 (3); D 170.9 (1), 71.6 (2), 39.6 (3), 24.5 (4), 21.5 (4-Me), 22.6 (5).

Cryptophycin 29
[α] _D +22.2 ^° (CHCl ₃ , c 1.13); UV λ _max (ε) 250 (17000), 284 (3300); IR (pure substance) ν _max 3415, 3272, 2960, 1744, 1734, 1674, 1504, 1259, 1197, 1174, 1067, 694 cm ^-1 ; EIMS m / z (relative intensity) 624/626 (2.6 / 1.1), 398/400 (44/15), 227 (100), 195/197 (50/16), 155/157 (59/20), 131 (63), 91 (95); high resolution EIMS m / z 624.2607 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₇ , - 0.5 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.75 (2, dd; 15.3 and 1.1), 6.69 (3, ddd; 15.3, 10.1 and 5.3), 2.36 (4, m), 2.54 (4, m), 5.03 (5, ddd; 11.0, 6.4 and 1.8), 2.56 (6, m ), 1.14 (6-Me, d; 6.8), 6.01 (7, dd; 15.8 and 8.8), 6.41 (8, d; 15.8), 7.28-7.33 (10/11/13/14, m), 7.22 (12, m); 3-chloro-4-methoxyphenylalanine (B) 4.76 (2, m), 5.67 (2-NH, d; 8.6), 3.0 (3, dd; 14.4 and 10.2), 3.14 (3, dd; 14.4 and 5.9), 7.22 (5, d; 2.2), 3.87 (7-OMe, s), 6.83 (8, d; 8.4), 7.08 (9, dd; 8.4 and 2.2); 3-aminopropionic acid (C) 2.55 (2-H ₂ , m), 3.44 (3, m), 3.55 (3, m), 6.89 (3-NH, br t; 5.7); leucine acid (D) 4.90 (2, dd; 9.9 and 3.5), 1.34 (3, ddd; 15.4, 10.3 and 3.5), 1.63 (3, m), 1.63 (4, m), 0.76 (4-Me, d ; 6.4), 0.72 (5, d; 6.4); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.6 (1), 125.2 (2), 141.5 (3), 36.4 (4), 77.1 (5), 42.3 (6), 17.3 (6-Me) 130.1 (7), 131.8 (8), 136.7 (9), 126.2 (10/14), 128.6 (11/13), 127.6 (12); B 170.9 (1), 53.8 (2), 34.9 (3), 129.9 (4), 131.0 (5), 122.4 (6), 153.9 (7), 56.1 (7-OMe), 112.2 (8), 128.4 ( 9); C 172.6 (1), 32.4 (2), 34.5 (3); D 170.4 (1), 71.5 (2), 39.7 (3), 24.4 (4), 21.2 (4-Me), 22.6 (5).

Cryptophycin 30
[α] _D -12.3 ^o (CHCl ₃ , s 1.53); UV λ _max (ε) 254 (17200), 284 (3600); IR (pure substance) ν _max 3414, 3306, 2961, 1738, 1729, 1660, 1504, 1258, 1205, 1183, 1066, 695 cm ^-1 ; EIMS m / z (relative intensity) 656/658 (1.0 / 0.3), 638/640 (3.0 / 1/0), 525/527 (3.8 / 1.3), 412/414 (10.5 / 3.6), 280/282 (10.3 / 3.8), 227 (29), 195/197 (48/17), 155/157 (74/21), 131 (100); high resolution EIMS m / z 656.2852 (calculated for С ₃₅ Н ₄₅ СlN ₂ O ₈ , 1.5 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 3,5-dihydroxy-6-methyl-8-phenyl-7-octenoic acid (A) 2.25 (2, dd; 16.0 and 9.6), 2.64 (2, brd; 16.0), 3.89 (3, m), 2.51 (3-OH, d; 6.4), 1.77 (4, ddd; 14.3, 9.8 and 2.1), 1.88 (4, ddd ; 14.3, 11.3 and 3.8), 4.88 (5, ddd; 11.3, 6.2 and 2.1), 2.53 (6, m), 1.10 (6-Me, d; 6.8), 5.99 (7, dd; 15.9 and 9.0), 6.40
(8, d; 15.9), 7.28-7.33 (10/11/13/14, m), 7.23 (12, t); 3-chloro-4-methoxyphenylalanine (B) 4.60 (2, m), 6.61 (2-NH, d; 8.1), 3.09 (3, dd; 14.2 and 5.6), 3.15 (3, dd; 14.2 and 7.3), 7.22 (5, d; 2.1), 3.86 (7-OMe, s), 6.83 (8, d; 8.3), 7.07 (9, dd; 8.3 and 2.1); 3-amino-2-methylpropionic acid (C) 2.67 (2, m), 1.21 (2-Me, d; 7.3), 3.26 (3, ddd; 13.6, 7.3 and 6.4), 3.63 (3, ddd; 13.6, 6.2 and 3.9), 6.75 (3-NH, br t; 6.3); leucic acid (D) 4.83 (2, dd; 9.6 and 4.1), 1.42 (3, m), 1.64 (3, m), 1.64 (4, m), 0.79 (4-Me, d; 6.4) 0.76 (5 , d; 6.4); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 171.6 (1), 42.4 (2), 66.0 (3), 41.3 (4), 76.0 (5), 42.0 (6), 17.3 (6-Me) 130.0 (7), 131.9 (8), 136.7 (9), 126.1 (10/14), 128.6 (11/13), 127.6 (12); B 170.8 (1), 54.3 (2), 35.1 (3), 130.1 (4), 131.1 (5), 122.2 (6), 153.8 (7), 56.1 (7-OMe), 112.1 (8), 128.7 ( 9); C 175.6 (1), 39.7 (2), 13.8 (2-Me), 41.5 (3); D 171.9 (1), 72.1 (2), 39.1 (3), 24.6 (4), 21.4 (4-Me), 22.7 (5).

Cryptophycin 31
[α] _D +50.6 ^° (MeOH, s 1.13); UV λ _max (ε) 242 (3800), 284 (700); IR (pure substance) ν _max 3412, 3272, 2961, 1745, 1725, 1678, 1537, 1481, 1270, 1196, 1176, 1000, 698 cm ^-1 ; EIMS m / z (relative intensity) 688/690 (1.2 / 1.0 / 0.4), 446/448/450 (7.9 / 6.7 / 3.1), 314/316/318 (17/11/3), 91 (100) ; high resolution EIMS m / z 688.2336 (calculated for C ₃₅ H ₄₂ Cl ₂ N ₂ O ₈ , -1.8 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 7.8-epoxy-5-hydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.78 (2, d; 15.5), 6.66 (3, ddd; 15.5, 9.4 and 6.0), 2.47 (4, ddd; 14.1, 10.8 and 9.4), 2.56 (4, m), 5.14 (5, ddd; 10.8, 4.7 and 1.7) , 1.82 (6, m), 1.15 (6-Me, d; 7.1), 2.93 (7, dd; 7.5 and 1.9), 3.70 (8, d; 1.9), 7.24-7.26 (10/14, m ), 7.34-7.39 (11/12/13, m); 3,5-dichloro-4-methoxyphenylalanine (B) 4.83 (2, m), 5.68 (2-NH, d; 9.0), 3.0 (3, dd; 14.4 and 7.3), 3.14 (3, dd; 14.4 and 5.6 ), 7.16 (5/9, s), 3.87 (7-OMe, s); 3-amino-2-methylpropionic acid (C) 2.74 (2, m), 1.22 (2-Me, d; 7.1), 3.20 (3, m), 3.58 (3, ddd; 13.5, 5.6 and 4.1), 6.82 (3-NH, br t; 5.6); leucinic acid (D) 4.83 (2, m), 1.38 (3, m), 1.72 (3, m), 1.72 (4, m), 0.87 (4-Me, d; 6.8), 0.86 (5, d; 6.8); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.4 (1), 125.4 (2), 141.0 (3), 36.7 (4), 76.3 (5), 40.6 (6), 13.5 (6-Me) , 63.0 (7), 58.9 (8), 136.7 (9), 125.6 (10/14), 128.7 (11/13), 128.6 (12); B 170.8 (1), 53.3 (2), 35.2 (3), 129.3 (4), 129.6 (5/9), 134.5 (6/8), 151.2 (7), 60.6 (7-OMe); C 175.3 (1), 38.3 (2), 13.9 (2-CH ₃ ), 41.5 (3); D 170.6 (1), 71.3 (2), 39.4 (3), 24.6 (4), 22.9 (4-Me), 21.3 (5).

Cryptophycin 40
[α] _D +41.6 ^° (CHCl ₃ , C 0.31); UV λ _max (ε) 242 (4974), 266 (3911), 274 (3666), 286 (2359), 328 (511); IR (pure substance) ν _max 3415, 2959, 1748, 1723, 1667, 1505, 1463, 1289, 1176 cm ^-1 ; EIMS m / z (relative intensity) 640/642 (5/2), 280/282 (7/3), 213 (13), 195/197 (51/17), 155 (29), 141 (32) 121 (28), 91 (100), 69 (47); high resolution EIMS m / z 640.2570 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₈ , -1.8 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7.8-epoxy-5-hydroxy-8-phenyl-2-octenoic acid (A) 5.77 (2, d; 15.1), 6.72 (3, ddd; 15.1, 9.7 and 4.9), 2.42 (4, m), 2.58 (4, m), 5.33 (5, m), 1.89 (6, ddd; 12.9, 8.1 and 5.0), 2.13 (6, ddd; 12.9, 9.3 and 5.0), 2.98 (7, ddd; 6.7, 4.5 and 1.9), 3.64 (8, d; 1.9), 7.31-7.39 (10/11/13/14, m) 7.22 (12, m); 3-chloro-4-methoxyphenylalanine (B) 4.83 (2, m), 5.64 (2-NH, d; 8.6), 3.03 (3, dd; 14.3 and 7.5), 3.14 (3, dd; 14.3 and 5.4), 7.21 (5, d; 2.0), 3.87 (7-OMe, s), 6.84 (8, d; 8.3), 7.08 (9, dd; 8.3 and 2.0); 3-amino-2-methylpropionic acid (C) 2.72 (2, m), 1.23 (2-Me, d; 7.3), 3.31 (3, dt; 13.8 and 6.9), 3.50 (3, ddd; 13.6, 5.7 and 3.9), 6.96 (3-NH, br t; 6.0); leucine acid (D) 4.85 (2, dd; 6.7 and 3.4), 1.42 (3, m), 1.72 (3, m), 1.72 (4, m), 0.86 (4-Me, d; 3.7), 0.87 (5, d; 3.7); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.3 (1), 125.2 (2), 140.9 (3), 39.0 (4), 72.0 (5), 37.3 (6), 59.0 (7), 58.7 (8), 140.9 (9), 125.6 (10/14), 128.7 (11/13), 128.5 (12); B 170.9 (1), 53.6 (2), 35.1 (3), 129.8 (4), 131.0 (5), 122.5 (6), 157.0 (7), 56.1 (7-OMe), 112.3 (8), 128.4 ( 9); C 175.6 (1), 38.3 (2), 14.1 (2-Me), 41.1 (3); D 170.9 (1), 71.4 (2), 39.4 (3), 24.5 (4), 21.5 (4-Me), 22.8 (5).

Cryptophycin 43
[α] _D +20 ^° (CHCl ₃ , s 0.2); UV λ _max (ε) 250 (20512), 282 (4083), 294 (1734); IR (pure substance) ν _max 3400, 3272, 2927, 1727, 1660, 1516, 1455, 1242, 1175 cm ^-1 ; EIMS m / z (relative intensity) 533 (24), 484 (3), 445 (14), 398 (9), 364 (29), 227 (59), 149 (67), 91 (100); high resolution EIMS m / z 590.3044 (calculated for C ₃₄ H ₄₁ N ₂ O ₇ , -5.2 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.75 (2, d; 15.3), 6.69 (3, ddd; 15.3, 9.9 and 5.3), 2.37 (4, dt; 14.2 and 10.4), 2.52 (4, m), 5.01 (5, ddd; 11.2, 6.4 and 1.8), 2.55 (6 , m), 1.13 (6-Me, d; 6.9), 6.01 (7, dd; 15.8 and 8.9), 6.41 (8, d; 15.8), 7.21-7.34 (10/11/12/13/14, m ); 4-methoxyphenylalanine (B) 4.80 (2, m), 5.64 (2-NH, d; 8.4), 3.06 (3, dd; 14.5 and 7.2), 3.13 (3, dd; 14.4 and 5.3), 7.06 (5 / 9, d; 8.4), 6.74 (6/8, d; 8.4); 3-amino-2-methylpropionic acid (C) 2.69 (2, m), 1.22 (2-Me, d; 7.3), 3.33 (3, m), 3.44 (3, dt; 14.0 and 4.7), 7.0 (3 -NH, m); leucine acid (D) 4.84 (2, dd; 10.0 and 3.6), 1.60-1.67 (3, m), 1.35 (3, m), 1.60-1.67 (4, m), 0.76 (5, d; 6.4), 0.73 (4-Me, d; 6.7); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 125.2 (2), 141.5 (3), 36.5 (4), 77.5 (5), 42.3 (6), 17.3 (6-Me), 130.1 (7) 131.8 (8), 136.8 (9), 126.2 (10/14), 128.6 (11/13), 127.6 (12); in 53.8 (2), 35.3 (3), 129.8 (4), 130.5 (5/9), 115.6 (6/8), 154.6 (7); C 38.3 (2), 14.1 (2-Me), 41.0 (3); D 71.6 (2), 39.6 (3), 24.5 (4), 21.2 (5), 22.9 (4-Me). Due to the small size of the sample, carbonyl carbon signals cannot be seen.

Cryptophycin 45
[α] _D +72.0 ^° (MeOH, s 0.12); UV λ _max (ε) 250 (25500), 284 (5300); IR (pure substance) ν _max 3407, 3239, 2958, 1743, 1727, 1667, 1538, 1469, 1242, 1196, 1177, 694 cm ^-1 ; EIMS m / z (relative intensity) 658/660/662 (2.1 / 1.4 / 0.3), 483 (7.6), 432/434/436 (9.5 / 6.4 / 1.8), 300/302/304 (8.0 / 5.5 / 1.2), 227 (100) 91 (87); high resolution EIMS m / z 658.2207 (calculated for C ₃₄ H ₄₀ Cl ₂ N ₂ O ₇ , 0.6 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.80 (2, d; 14.7), 6.66 (3, ddd; 14.7, 8.5 and 5.5), 2.38 (4, m), 2.53 (4, m), 4.97 (5, br dd; 10.4 and 6.2), 2.57 (6, m), 1.14 (6-Me, d; 6.7), 6.01 (7, dd; 15.9 and 8.7), 6.42 (8, d; 15.9), 7.28-7.34 (10/11/13/14, m), 7.22 (12; m ); 3,5-dichloro-4-hydroxyphenylalanine (B) 4.82 (2, m), 5.73 (2-NH, br d; 8.7), 3.02 (3, dd; 14.3 and 6.2), 3.10 (3, dd; 14.3 and 5.2), 7.14 (5/9, s), 5.79 (7-OH, s); 3-amino-2-methylpropionic acid (C) 2.73 (2, m), 1.21 (2-Me, d; 7.0), 3.17 (3, m), 3.60 (3, m), 6.81 (3 -NH, br t; 6.7); leucine acid (D) 4.84 (2, dd; 10.0 and 3.2), 1.38 (3, ddd; 14.9, 10.2 and 3.2), 1.65 (3, m), 1.65 (4, m), 0.78 (4-Me, d ; 6.5), 0.73 (5, d; 6.5); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.5 (1), 125.4 (2), 141.2 (3), 36.4 (4), 77.6 (5), 42.3 (6), 17.3 (6-Me) 130.0 (7), 131.9 (8), 136.7 (9), 126.2 (10/14), 128.6 (11/13), 127.6 (12); B 171.0 (1), 53.2 (2), 35.0 (3), 130.4 (4), 129.1 (5/9), 121.0 (6/8), 146.7 (7); C 175.2 (1), 38.5 (2), 13.9 (2-Me), 41.6 (3); D 170.7 (1), 71.5 (2), 39.5 (3), 24.6 (4), 22.7 (4-Me), 21.2 (5).

Cryptophycin 49
[α] _D +68.1 ^° (MeOH, s 0.075); UV λ _max (ε) 246 (25500), 284 (5200); IR (pure substance) ν _max 3401, 3282, 2962, 1744, 1728, 1668, 1540, 1505, 1464, 1258, 1198, 1177, 1066, 694 cm ^-1 ; EIMS m / z (relative intensity) 624/626 (0.8 / 0.3), 398/400 (43/14), 227 (78), 195/197 (58/26) 91 (100); high resolution EIMS m / z 624.2650 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₇ , -4.8 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octanoic acid (A) 5.77 (2, d; 14.1), 6.67 (3, m), 2.38 (4, m), 2.50 (4, m), 5.01 (5, m), 2.56 (6, m), 1.13 (6-Me, d; 6.5), 6.03 (7, dd; 15.8 and 8.6), 6.42 (8, d; 15.8), 7.29-7.35 (10/11/13/14, m), 7.23 (12; m); 3-chloro-4-methoxyphenylalanine (B) 4.82 (2, m), 5.64 (2-NH, m), 3.06 (3, m), 3.13 (3, m), 7.22 (5, m), 3.87 (7 -OMe, s), 6.83 (8, m), 7.08 (9, m); 3-amino-2-methylpropionic acid (C) 2.72 (2, m), 1.22 (2-Me, d; 6.7), 3.26 (3, m), 3.53 (3, m), 6.90 (3-NH, m ); 2-hydroxyvaleric acid (D) 4.81 (2, dd; 8.8 and 3.9), 1.63 (3, m), 1.68 (3, m), 1.33 (4-H ₂ , m), 0.74 (5, t; 7.3) .

Cryptophycin 50
[α] _D + 32.0 ^° (CHCl ₃ , p. 0.44); UV λ _max (ε) 242 (4933), 262 (3996), 274 (3719), 286 (2430), 332 (359); IR (pure substance) ν _max 3412, 3274, 2958, 1752, 1724, 1676, 1648, 1503, 1465, 1258, 1177, 1066, 753 cm ^-1 ; EIMS m / z (relative intensity) 640/642 (4/2), 398/400 (11/4), 280/282 (10/3), 227 (17), 195/197 (57/18), 157 (20), 141 (31), 91 (100); high resolution EIMS m / z 640.2531 (calculated for C ₃₄ H ₄₁ ClN ₂ O ₈ , 2.1 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 7,8-epoxy-5-hydroxy-6-methyl-8-phenylloctanoic acid (A) 5.73 (2, d; 15.7), 6.67 (3, ddd; 15.7, 9.7 and 5.4), 2.45 (4, m), 2.55 (4, m), 5.13 (5, ddd; 11.2, 5.0 and 1.7), 1.78 (6, m), 1.15 (6-Me, d; 6.9), 2.91 (7, dd; 7.5 and 1.9), 3.68 (8, d; 1.7), 7.25 (10/14, m), 7.33-7.38 (11/12/13; m); 3-chloro-4-methoxyphenylalanine (B) 4.80 (2, ddd; 8.3, 7.1 and 5.4), 5.61 (2-NH, d; 8.3), 3.03 (3, dd; 14.4 and 7.3), 3.13 (3, dd ; 14.4 and 5.6), 7.21 (5, d; 1.9), 3.87 (7-OMe, s), 6.83 (8, d; 8.4), 7.07 (9, dd; 8.4 and 2.2); 3-amino-2-methylpropionic acid (C) 2.71 (2, m), 1.22 (2-Me, d; 7.3), 3.29 (3, dt; 13.6 and 6.9), 3.49 (3, ddd; 13.6, 6.7 and 5.0), 6.92 (3-NH, br t; 6.7); 2-hydroxypentanoic acid (D) 4.75 (2, dd; 9.2 and 3.7), 1.55 (3, m), 1.65 (3, m), 1.33 (4-H ₂ , m), 0.84 (5, t; 7.3) ; ¹³ C NMR (CDCl ₃ ) unit δ values (carbon position) A 165.3 (1), 125.3 (2), 141.0 (3), 36.9 (4), 76.3 (5), 40.8 (6), 13.6 (6-Me ), 63.2 (7), 59.1 (8), 136.8 (9), 125.5 (10/14), 128.7 (11/13), 128.5 (12); B 170.9 (1), 53.6 (2), 35.1 (3), 129.8 (4), 131.0 (5), 122.5 (6), 154.0 (7), 56.1 (7-OMe), 112.3 (8), 128.5 (9); C 175.6 (1), 38.4 (2), 14.1 (2-Me), 41.2 (3); D 170.4 (1), 72.4 (2), 32.7 (3), 18.4 (4), 13.5 (5).

Cryptophycin 54
EIMS m / z (relative intensity) 654/656 (17/10), 493 (5), 411/413 (12/4), 280 (16), 227 (25), 195/197 (45/25) , 141 (30), 91 (100); high resolution EIMS m / z 654.2686 (calculated for C ₃₅ H ₄₃ ClN ₂ O ₈ , 2.2 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) (A) 5.73 (2, d; 15.4), 6.66 (3, ddd; 15.4, 9.7 and 5.7), 2.46 ( 4, m), 2.53 (4. m), 5.16 (5, ddd; 11.0, 4.2, 1.7), 1.79 (6, m), 1.14 (6-Me, d; 6.8), 2.89 (7, dd ; 7.4 and 1.8), 3.69 (8, d; 1.9), 7.25 (10/14, m), 7.30-7.38 (11/12/13, m); (B) 4.81 (2, m), 5.63 (2-NH, d; 8.6), 3.03 (3, dd; 14.5, 7.3), 3.13 (3, dd; 14.5, 5.5), 7.21 (5, d; 2.2 ), 3.87 (7-OMe, s), 6.83 (8, d; 8.4), 7.07 (9, dd; 8.4 and 2.2); (C) 2.73 (2, m), 1.22 (2-Me, d; 7.3), 3.26 (3, ddd; 13.4, 6.8, 6.8), 3.51 (3, ddd; 13.4, 6.8, 5.3), 6.88 (3 -NH, br t; 6.8); (D) 4.73 (2, d; 4.2), 1.78-1.82 (3, m), 0.92 (3-Me, d; 6.8), 1.36-1.41 (4, m), 1.18-1.20 (4, m), 0.80 (5, t; 7.5); ¹³ C NMR (CDCl ₃ ) unit δ (carbon position) A 165.3 (1), 125.4 (2), 141.0 (3), 36.6 (4), 76.3 (5), 40.6 (6), 13.2 (6-Me) , 63.1 (7), 58.7 (8), 136.7 (9), 125.4 (10/14), 128.6 (11/13), 128.5 (12); B 170.9 (1), 53.5 (2), 35.0 (3), 129.8 (4), 131.0 (5), 125.2 (6), 153.9 (7), 56.1 (7-OMe), 112.2 (8), 128.4 (9); C 175.4 (1), 38.5 (2), 14.0 (2-Me), 41.3 (3); D 169.4 (1), 76.5 (2), 36.1 (3), 15.6 (3-Me), 24.0 (4), 11.2 (5).

Example 6. Synthesis of Cryptophycin Derivatives
Cryptophycin 8
To a solution of 3.8 mg of Cryptophycin 1 in 1.5 ml of a 2: 1 mixture of 1,2-dimethoxyethane / water was added 9 μl of 1N Hcl. The solution was allowed to mix for 4 hours, neutralized with potassium carbonate and evaporated. The residue was in the form of a partition between water and CH ₂ Cl ₂ , CH ₂ Cl ₂ - the soluble part was purified using reverse-phase liquid chromatography by gel to obtain 3.3 mg of pure Cryptophycin 8.

(Hereinafter in all subsequent examples)
Data from EIMS electron emission spectroscopy m / z (relative intensity) 690/692/694 (0.8 / 0.5 / 0.2). High resolution EIMS m / z 690.2533 (calculated for C ₃₅ H ₄₄ Cl ₂ N ₂ O ₈ -5.8 mmu error). ¹ H-NMR (CDCl ₃ ) - data of proton magnetic resonance; amino or hydroxy-acid unit δ (carbon position, multiplicity, J in Hz) 8-chloro-5,7-dihydroxy-6-methyl-8-phenyl-2-octenoic acid (A) 5.79 (2, d - doublet; 15.4) , 6.69 (3, ddd - triple doublet; 15.4, 9.7 and 5.6), 2.68 (4, ddt - double doublet triplet; 14.0, 5.5 and 1.8), 2.38 (4, m - multiplet); 5.11 (5, ddd - triple doublet; 10.8, 8.6 and 1.8), 2.51 (6, m - multiplet), 1.05 (6-Me, d - doublet; 7.0), 4.01 (7dd - double double; 9.6 and 1.4), 4.65 (8, d - doublet; 9.6), 7.36-7.41 (10/11/12/13/14, m) leucinic acid (D) 4.92 (2, dd; 10.1 and 3.5), 1.76 (3/4, m ), 1.45 (3, m), 0.94 (5, d; 6.6), 0.94 (4-Me, d; 6.4); 3-amino-2-methylpropionic acid (C) 2.73 (2, m), 1.22 (2-Me, d; 7.2), 3.25 (3, ddd; 13.6, 6.8 and 6.1), 3.54 (3, ddd; 13.5, 6.1 and 3.4), 6.91 (3-NH, brt; 6.1); 3-chloro-4-methoxyphenylalanine (B) 4.82 (2, ddd; 8.8, 7.2 and 6.6), 5.64 (2NH, d; 8.8), 3.03 (3, dd; 15.4 and 7.2), 3.16 (3, dd; 15.4 and 5.6), 7.23 (5, d; 2.2), 3.88 (7-OCH ₃ , s), 6.85 (8, d; 8.5), 7.09 (9, dd; 8.5 and 2.2).

Cryptophycin 9
To a solution of 10 mg of Cryptophycin 1 in 1 ml of dry methanol was added 10 μl of HCl (obtained by processing 1.25 g of thionyl chloride with 25 ml of MeOH). After stirring for 4 hours, the solvent was removed in vacuo and the sample was left in vacuo for 12 hours. Purification by reverse phase liquid gel chromatography gave 8 mg of pure Cryptophycin 9.

¹ H NMR (CDCl ₃ ): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 5,7-dihydroxy-8-methoxy-6-methyl-8-phenyl-2-octanoic acid (A) 5.76 ( 2, d; 15.5), 6.67 (3, ddd; 15.5, 9.5 and 5.6), 2.34 (4, ddd; 14.1, 11.1 and 9.5), 2.62 (4, dddd; 14.1, 5.6, 1.8 and 1.5), 5.09 ( 5, ddd; 11.1, 7.8 and 1.8), 2.24 (6, dqd; 7.8, 7.0 and 2.2), 1.03 (6-Me, d; 7.0), 3.71 (7, dd; 8.3 and 2.2), 4.03 (8, d; 8.3), 3.20 (8-OCH ₃ , s), 7.31-7.40 (10/11/12/13/14, m); leucine acid (D) 4.86 (2, dd; 9.8 and 3.5), 1.71 (3/4, m), 1.41 (3, m), 0.89 (5/4-Me, d; 6.4); 3-amino-2-methylpropionic acid (C) 2.71 (2, ddq; 6.8, 3.9 and 7.2), 1.21 (2-Me, d; 7.2), 3.23 (3, ddd; 13.5, 6.8 and 6.0), 3.52 ( 3, ddd; 13.5, 6.0 and 3.9), 6.90 (3-NH, brt; 6.0), 3-chloro-4-methyloxyphenylamine (B) 4.82 (2, ddd; 8.8, 7.4 and 5.7), 5.66 (2 -NH, d; 8.8), 3.02 (3, dd; 14.4, 7.4), 3.15 (3, dd; 14.4 and 5.5), 7.23 (5, d; 2.2), 3.87 (7-OCH ₂ , s), 6.84 (8, d; 8.5), 7.08 (9, dd; 8.5 and 2.2).

Cryptophycin 10
To a stirred solution of 7 mg Cryptophycin 9 in 1 ml of acetone and 0.3 ml of water was added 8 μl of a 2 N NaOH solution. After stirring for 4 hours, the solution was neutralized to pH 7 with 1 N HCl and the solvent was removed under reduced pressure. The residue was subjected to reverse phase liquid chromatography-gel chromatography using a 7: 3 MeOH / H ₂ O mixture to give pure Cryptophycin 10 (5 mg).

¹ H NMR (CD ₂ OD): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 5,7-dihydroxy-8-methoxy-6-methyl-8-phenyl-2-octanoic acid (A) 5.99 (2, dt; 15.4 and 1.3), 6.82 (3, dt; 15.4 and 7.3), 2.30 (4, m), 2.50 (4, m), 3.66 (5, td; 7.8 and 3.5), 2.05 ( 6, d; 1.8, and 7.0), 0.96 (6-Me, d; 7.0), 4.04 (7, dd; 8.8 and 2.0), 4.01 (8, d; 8.8), 3.12 (8-OCH ₃ , s) 7.26-7.36 (10/11/12/13/14, m); 3-amino-2-methylpropionic acid (C) 2.50 (2, m), 1.02 (2-Me, d; 7.3), 3.16 (3, dd; 13.4 and 6.9), 3.82 (3, dd; 13.4 and 6.6); 3-chloro-4-methyloxyphenylamine (B) 4.57 (2, dd; 8.5 and 6.5), 2.82 (3, dd; 13.9 and 8.6), 3.03 (3, dd; 13.9 and 6.5), 7.25 (5, d ; 2.2), 3.82 (7-OCH ₃ , s), 6.96 (8, d; 8.6), 7.13 (9, dd; 8.6 and 2.2). ¹³ C NMR (CD ₃ OD): δ 179.5, 173.4, 168.2, 155.4, 143.7, 141.7, 131.9, 131.7, 129.8, 129.3, (2C), 129.2 (2C), 128.8, 126.2, 123.2, 113.4, 85.9, 74.5 , 74.1, 56.8, 56.6, 56.3, 43.3, 41.2, 40.2, 38.8, 38.0, 15.5, 9.9.

Cryptophycin 12
To a solution of 5 mg of Cryptophycin 1, 5 or 8 in 1 ml of a 4: 1 acetone / water mixture was added 15 μl of a 2 N NaOH solution. After stirring at room temperature for 5 hours, the reaction mixture was neutralized to pH 7 with 1 N HCl and the solvent was evaporated. A substance soluble in CH ₂ Cl _{2 was} passed through a small silicon cartridge with CH ₂ Cl ₂ , 1: 1 EtOAc / CH ₂ Cl ₂ , and EtOAc. The fraction eluted with EtOAc contained pure Cryptophycin 12.

¹ H NMR (CD ₃ OD): amino or hydroxy-acid unit δ (carbon position, multiplicity; J in Hz) 5,7,8-trihydroxy-6-methyl-8-phenyl-2-octanoic acid (A) 6.07 (2 , ddd; 15.5, 1.3 and 1.2), 6.40 (3, dt; 15.5 and 7.3), 2.49 (4, m), 2.60 (4, m), 3.92 (5, ddd; 9.3, 6.7 and 4.5), 1.94 ( 6, m), 1.07 (6-Me, d; 6.6), 3.61 (7, dd; 8.9 and 7.6), 4.56 (8, d; 7.6), 7.36 (10/14, dd; 7.4 and 1.5), 7.32 (11/13, brt; 7.5), 7.25 (12, m); 3-amino-2-methylpropionic acid (C) 2.54 (2, ddq; 7.0, 6.6 and 7.0), 1.02 (2-Me, d; 7.0), 3.14 (3, dd; 13.5 and 7.0), 3.42 ( 3, dd; 13.4 and 6.6); 3-chloro-4-methyloxyphenylamine (B) 4.57 (2, dd; 8.4 and 6.7), 2.83 (3, dd; 13.8 and 8.4), 3.02 (3, dd; 13.8 and 6.6), 7.25 (5, d; 2.1 ), 3.82 (7-OCH ₃ , s), 6.95 (8, d; 8.5), 7.12 (9, dd; 8.5 and 2.1). Methylation of Cryptophycin 12 with diazomethane gave Cryptophycin 6.

Cryptophycin 14
To a solution of 3 mg Cryptophycin 6 in 1 ml of a 3: 1 acetone / water mixture was added 5 μl of a 2 N NaOH solution. After stirring for 5 hours, the reaction mixture was neutralized to pH 7 with 1 N HCl and then evaporated to dryness. The residue was subjected to reverse-phase liquid chromatography-gel chromatography to obtain 2.4 mg of Cryptophycin 14,
¹ H NMR (CD ₃ OD): amino or hydroxy acid unit δ (carbon position, multiplicity; J in Hz) 5-hydroxy-6-methyl-8-phenyl-2,7-octadiene acid (A) 5.98 (2, d ; 15.3), 6.78 (3, dt; 15.3 and 7.5), 2.35 (4, m), 3.64 (5, td; 7.2 and 4.8), 2.47 (6, m), 1.14 (6-Me, d; 6.9) , 6.22 (7, dd; 15.9 and 8.1), 6.39 (8, d; 15.9), 7.24-7.36 (10/11/12/13/14, m), 3-amino-2-methylpropionic acid (C) 2.35 (2, m), 1.02 (2-Me, d; 6.9), 3.18 (3, dd; 13.2 and 6.6), 3.36 (3, dd; 13.2 and 4.5); 3-chloro-4-methyloxyphenylalanine (B) 4.58 (2, dd; 8.7 and 6.3), 2.80 (3, dd; 13.8 and 9.0), 3.05 (3, dd; 13.8 and 6.3), 7.25 (5, d ; 2.1), 3.82 (7-OCH ₃ , s), 6.95 (8, d; 8.4), 7.13 (9, dd; 8.4 and 2.1).

Cryptophycin 35
A catalytic amount of PtO ₂ was added to the flask containing 0.5 ml of CH ₂ Cl ₂ . The flask was evacuated, H ₂ was introduced and the mixture was stirred at room temperature for 20 minutes. A solution of 10 mg of Cryptophycin 1 in a minimum amount of CH ₂ Cl ₂ was added and the mixture was stirred at room temperature for 45 minutes. The catalyst was removed by filtration through celite / cotton and the solvent was evaporated. Treatment of the residue with reverse phase liquid chromatography using a C18 column afforded 6.5 mg of Cryptophycin 35.

EIMS m / z (relative intensity) 656/658 (25/10), 412/414 (25/12), 280/282 (20/10), 195/197 (78/25), 141 (58), 91 (100); high resolution EIMS m / z 656.2864 (calculated for C ₃₅ H ₄₅ ClN ₂ O ₈ , 0.0 mmu error); ¹ H NMR (CDCl ₃ ): amino or hydroxy acid unit δ values (carbon position, multiplicity; J in Hz) 2,3-dihydro-7,8-epoxy-5-hydroxy-6-methyl-8-phenyl octanoic acid ( A) 2.32 (2, ddd; 14.5, 9.2, 5.8), 2.10 (2, ddd; 14.5, 9.2, 6.2), 1.5-1.8 (3/4 overlay m), 5.07 (5, ddd; 12.5, 5.6 , 2.0), 1.80 (6, m), 1.12 (6-Me, d; 7.0), 2.90 (7, dd; 7.4, and 1.8), 3.67 (8, d; 1.8), 7.24 (10/14, m ), 7.32-7.38 (11/12/13, m); 3-chloro-4-methoxyphenylalanine (B) 4.71 (2, ddd; 8.7, 6.4, 6.3), 5.62 (2-NH, d; 8.7), 3.08 (2H-3, br d; 6.4), 7.19 (5, d; 2.0), 3.87 (7-OMe, s), 6.83 (8, d; 8.5), 7.07 (9, dd; 8.4, 2.0); 3-amino-2-methylpropionic acid (C) 2.72 (2, m), 1.18 (2-Me, d; 6.9), 3.12 (3, ddd; 11.4, 10.6, 5.6), 3.70 (3, ddd), 6.76 (3-NH, br t; 6.0); leucine acid (D) 4.83 (2, dd; 9.9, 3.8), 1.39 (3, m), 1.70 (3, m), 1.72 (4, m), 0.87 (4-Me, d; 5.3), 0.86 ( 5, d; 5.3); ¹³ C NMR (CDCl ₃ ) unit δ values (carbon position) A 172.4 (1), 36.2 (2), 32.0 (3), 21.1 (4), 76.6 (5), 40.2 (6), 13.6 (6-Me ), 63.3 (7), 59.2 (8), 136.8 (9), 125.6 (10/14), 128.7 (11/13), 128.6 (12); B 170.7 (1), 53.7 (2), 35.5 (3), 130.0 (4), 131.1 (5), 122.2 (6), 153.8 (7), 56.1 (7-OMe), 112.1 (8 ), 128.5 (9); C 175.2 (1), 38.2 (2), 13.6 (2-Me), 42.1 (3); D 171.9 (1), 71.7 (2), 39.6 (3), 24.5 (4), 22.9 (4-Me), 21.4 (5).

 Example 7. Analysis of the activity of cryptocin compounds in microtubule depolymerization.

 Vinblastine, cytochalazine B, tetramethylrodamine isothiocyanate (TRITC) -phaloidin, sulforodamine B (SRB), and anti-β-tubulin and vimentin antibodies were obtained from Sigma Chemical Company. Eagle's basal medium containing Earle salts (BME) was from Gibco and fetal bovine serum (FBS) was purchased from Halcone Laboratories.

Cell lines
The Jurkat T line of rat aortic smooth muscle leukemia cells and A-10 cells was obtained from the American Type Culture Collection and cultured in BME containing 10% FBS and 50 μg / ml gentamicin sulfate. Ovarian human carcinoma cells (SKOV3) and the sub-line selected for resistance to vinblastine (SKVLB1) were a generous gift from Dr. Victor Ling of the Ontario Cancer Institute. Both cell lines were contained in BME containing 10% FBS and 50 μg / ml gentamicin sulfate. Vinblastine was added at a final concentration of 1 μg / ml to SKVLB1 cells 24 hours after passage to maintain the selected pressure for P-glycoprotein overexpressing cells.

 Cell proliferation and cycle delay assays.

Cell proliferation assays were performed as described by Skehan et.al. ¹¹ . For Jurkat cells, cultures were treated with incubated drugs as described in Skehan et.al. ¹¹ , and total cell sets were determined by counting cells in a hemacytometer. The percentage of cells in mitosis was determined by staining with 0.4% Giemsa in PBS followed by three fast washes with PBS. At least 1000 cells per treatment were counted for the presence of myotic structures and a myotic index calculated as the ratio of cells with myotic structures to the total number of cells counted.

 The effect of cryptophycin and vinblastine on the proliferation of Jurkat cells and the cell cycle.

Dose response curves for the effects of cryptophycin compounds and vinblastine on cell proliferation and the percentage of cells in mitosis are shown in figures 2A and 2B, respectively. Less than 3% of untreated cells exhibit mitotic structures. Both cryptophycin compounds and vinblastine cause a dose-dependent increase in the percentage of cells observed in mitosis. An increase in the mitotic index closely correlated with a decrease in cell proliferation, i.e. the concentrations of both cryptophycin compounds and vinblastine, which caused 50% of the cells to accumulate in mitosis, were actually the same as the concentration that inhibited cell proliferation by 50%. IC ₅₀ values for cryptophycin compounds and vinblastine for these effects were 0.2 and 8 nM, respectively.

The effect of cytochalazine B. vinblastine and cryptophycin on the cytoskeleton
Aortic smooth muscle cells (A-10) were grown on glass coverslips and treated with PBS, 2 μM cytochalazine B, 100 nM vinblastine or 10 nM cryptophycin compounds. After 24 hours, microtubule and vimentin intermediate fibers were visualized using indirect immunofluorescence analysis and microfibers were stained using TRITC-phalloidin. The morphological effects of each drug were investigated. The untreated cells showed completely expanded microtubule networks with microtubule organizing centers surrounding the nucleus. Vimentin intermediate fibers were also uniformly distributed throughout the cytoplasm, while bundles of microfibers were concentrated along the major axis of the cell. Cytochalazine B caused complete depolymerization of microfibers along with the accumulation of paracrystalline residues. This compound did not affect the distribution of microtubules, nor the distribution of intermediate fibers. Both vinblastine and the cryptophycin compound caused a marked emptying of microtubules. None of the compounds influenced the organization of microfibers; however, vimentin intermediate fibers flattened to form concentric rings around the nucleus of cells treated with either vinblastine or a cryptophycin compound.

The effect of cryptophycin and vinblastine on taxol-stabilized microtubules
A-10 cells were treated for 3 hours with 0 or 10 μM Taxol prior to the addition of PBS, 100 nM vinblastine, or 10 nM cryptophycin compound. After 24 hours, the organization of microtubules was investigated using immunofluorescence analysis, as described above. Compared to control cells, microtubules in taxol-treated cells were intensively connected, especially in the polar cell regions. As before, vinblastine caused the complete depolymerization of microtubules in untreated pre-cells. However, taxol pretreatment prevented microtubules from depolymerization in response to vinblastine. Similarly, taxol pretreatment completely stabilized microtubules against cryptophycin-induced depolymerization.

Reversibility of microtubule depolymerization due to vinblastine and cryptophycin
A-10 cells were treated with either 100 nM vinblastine or 10 nM cryptophycins for 24 hours, resulting in complete microtubule depolymerization. Then the cells were washed and incubated in a drug-free medium for 1 hour or 24 hours. Microtubules underwent rapid re-polymerization after removal of vinblastine, showing significant microtubule levels after 1 hour and complete morphological recovery in 24 hours. In contrast, microtubules do not reappear in cells treated with cryptophycin compounds, either 1 hour or 24 hours after removal of the compound.

Reversibility of Inhibition of Cell Proliferation by Cryptophycin, Vinblastine, and Taxolam
SKOV3 cells were treated for 24 hours with previously established IC ₅₀ doses of vinblastine, cryptophycin compounds or taxol (i.e., the values determined in the experiments summarized in Table 5). During this time, the cell density increased from 0.4 to 0.5 ± 0.05 absorption units (Figure 3), indicating a 25% increase in the number of cells for all three treatments. Removal of the drug led to the rapid growth of the cells treated with vinblastine, so that their number increased approximately three times within 24 hours. In contrast, cells treated with cryptophycin compounds or taxol remained blocked, increasing only 0.2-0.4 times within 24 hours after drug removal. The proliferative capacity of cells treated with cryptophycin or taxol was subsequently restored, as the cells then doubled in the next 24 hours.

The effect of combinations of vinblastine and cryptophycin on cell proliferation
SKOV3 cells were treated with combinations of cryptophycin and vinblastine for 48 hours. Then, the percentage of surviving cells was determined and IC ₅₀ values were calculated for each combination. The effect of these treatments with the indicated combinations, as well as treatments with one drug, is depicted as isobolograms (Figure 4). The IC ₅₀ values for combinations of cryptophycin compounds and vinoblastin very close to the line of additivity, indicating that these two drugs induce only additive inhibition of cell proliferation.

Toxicity of cryptophycin, vinblastine and taxol to SKOV3 and SKVLBI cells
SKVLBI cells resist natural anti-cancer drugs due to their over-expression of P-glycoprotein ¹² . The ability of taxol, vinblastine and cryptophycin compounds to inhibit the growth of SKOV3 and SKVLBI cells is summarized in Table 5. Taxol induces dose-dependent inhibition of proliferation of both cell lines with IC ₅₀ values for SKOV3 and SKVLBI cells 1 and 8000 and M, respectively. Vinblastine also inhibits the growth of both cell lines, with IC ₅₀ values of 0.35 and 4200 nM for SKOV3 and SKVLBI cells, respectively. Cryptophycins show IC ₅₀ values of 7 and 600 pM for SKOV3 and SKVLBI cells, respectively. Resulting resistance factors for SKVLBI cells to compounds are calculated as IC ₅₀ for SKVKBI. IC ₅₀ for SKOV3 cells are also indicated in table 5.

 Thus, it demonstrates that the present invention provides new cryptophycin compounds, as well as the previously described cryptophycin compounds, which are potent cell proliferation inhibitors acting by breaking microtubule networks and inhibiting mitosis. Cryptophycin compounds disrupt microtubule organization and thus normal cellular functions, including mitosis.

 Classical anti-microtubule agents, such as colchicine and Wink alkaloids, delay cell division during mitosis. It seems appropriate to compare the effect of one of these agents on cell proliferation with cryptophycin compounds. For this purpose, vinblastine of the Wink alkaloid was selected as the representative of antimicrotubule agents. Thus, a comparison was made of the effect of cryptophycin compounds and vinblastine on the proliferation and development of the cell cycle of the Jukart T cell leukemia cell line. Both compounds caused a parallel dose-dependent inhibition of cell proliferation and cell accumulation in mitosis.

 Since antimitotic effects are usually mediated by the destruction of microtubules in mitotic spindles, the effects of cryptophycin compounds on cytoskeletal structures have been characterized using fluorescence spectroscopy. Immunofluorescence staining of cells treated with either a cryptophycin compound or vinblastine clearly demonstrates that both compounds cause complete loss of microtubules. Similar studies with SKOV3 cells demonstrate that the anti-microtubule effects of cryptophycin compounds are not unique to the smooth muscle cell line. No drug has an effect on microfiber bundle levels or distribution, which was readily induced by cytochalazine B, indicating that microtubule loss cannot be due to a nonspecific mechanism, such as activation of proteases or loss of energy charge. Both vinblastine and cryptophycin compounds also markedly accelerate the collapse of vimentin intermediate fibers so that brightly colored rings form around the cell nuclei.

 Removing vinblastine from the culture medium leads to rapid re-polymerization of microtubules. In contrast, cells treated with cryptophycin compounds remained depleted with respect to microtubules for at least 24 hours after the compound was removed from the cultures.

The present invention demonstrates that cryptophycin compounds disrupt P-glycoprotein-mediated multiple drug resistance. Transport due to P-glycoprotein limits the ability of anticancer drugs of the natural product to inhibit the growth of tumor cells with the required or de novo drug resistance ^13-15 . Wink alkaloids, although very useful as an initial source of chemotherapy, are extremely good medications for the transport of P-glycoprotein and are therefore very limitedly useful against P-glycoprotein-mediated MDR tumors.

 Therefore, the identification of agents that can block multiple drug resistance should lead to the development of useful and new anti-cancer agents. The cryptophycin compounds of the present invention are such agents since they are poor substrates for P-glycoprotein-mediated transport. This fact is reflected in the low factor of cellular resistance for cryptophycin compounds, in comparison with vinblastine, taxol and other natural medicinal products.

 Complete synthesis of cryptophycins.

 The structures of the newly synthesized compounds, namely Cryptophycin 51, 52, 53, 55, 56, 57, 58, and 61, were directly confirmed using a methodology that is well known to those skilled in the art. Mass spectral data were consistent with molecular compositions. The proton and carbon data of the NMR spectra were similar to those for cryptophycin 1 and the corresponding analogues of natural origin and semisynthetic analogues.

 The following examples demonstrate the complete synthesis of cryptophycin compounds, as well as their use as therapeutic agents in accordance with the invention.

Example 8. Synthesis of Cryptophycin 51
S-trans-3-penten-2-ol (A)
A mixture of racemic trans-3-penten-2-ol (933 mg, 11 mmol), trifluoroethyl laurate (4.14 g, 15 mmol) and pork pancretite lipase (PPL, 2.0 g) in 25 ml of anhydrous diethyl ether was stirred for 80 hours . Then the PPL was filtered and washed three times. The ether filtrates were evaporated and the sticky oil was then vacuum distilled with a short reflux condenser. S-trans-3-penten-2-ol (A) was condensed in a trap cooled with liquid nitrogen (338 mg).

¹ H-NMR (CDCl ₃ ) δ 5.57 (4-H; dq, -15.3 / 6.0). 5.47 (3-H; ddd, -15.3 / 6.4 / 1.2), 4.19 (2-H; 1: 4: 6: 4: 1 pentuplet. 6.4). 2.24 (OH; bs), 1.63 (5-H ₃ ; d. 6.0), 1.19 (1-H ₃ ; d, 6.4). ¹³ C NMR (CDCl ₃ ) δ 135.5 (3), 125.5 (4). 68.7 (2), 23.3 (5), 17.5 (1).

S-trans-2- (2-propynyloxy) -3-pentene (B)
To a vigorously stirred mixture of S-enantiomer A (628 mg, 7.3 mmol), tetrabutylammonium hydrosulfate (138 mg, 0.41 mmol) and 40% NaOH in water (5 ml) at 0 ^° C., propargyl chloride (767 mg, 10.3 mmol) was added dropwise. , 745 μl), Intensive stirring was continued overnight, after which the mixture was neutralized with HCl at 0 ^° C and the propargyl ether was extracted into pentane. The extract was evaporated and the propargyl ether was purified on a short silica gel column (2% diethyl ether / pentane) to give 778 mg of propargyl ether B,
[α] _D -118.9 ^o (s 2.0, CHCl ₃ ): ¹ N NMR (CDCl ₃ ) δ 5.70 (4-H; dq, 18.5 / 6.5), 5.31 (3-H; ddd, 18.5 / 7.2 / 1.4) , 4.15 (1'-H; dd, -15.6 / 2.1), 4.01 (1'-H; dd, -15.6 / 2.1), 4.01 (2-H; m), 2.38 (3'-H; t, 2.1 ), 1.73 (5-H; dd, 6.5 / 1.4). 1.25 (1H; d, 6.3).

(3R, 4R) -4-Methylhept-5 (E) -en-1-in-3-ol (C)
An aliquot of a hexane solution of butyl lithium (2.5 M, 5.1 ml, 12.8 mmol) was evaporated in vacuo and the residue was cooled to -90 ^° C. A solution of propargyl ether B (454 mg, 3.66 mmol) in 10 ml of THF was slowly added. After that allow the temperature to rise overnight to room temperature, the reaction mixture is quenched with a solution of NH ₄ Cl. Extraction with ether three times, evaporation of the dried extract and purification of the residue on a column of silica gel (:% EtOAc / hexane) gives 332 mg of alcohol C (71% yield).

131.5 (5), 127.9 (6), 83.5 (2), 73.6 (1), 66.2 (3), 43.4 (4), 18.1 (7), 15.7 (4-Ме).[α] _D + 32.9 ^° (c 3.0, CHCl ₃ ); IR (NaCl) ν _max 3306, 2968, 1455, 1379, 1029, 975 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ 5.61 (6-H; dq, 15.3 / 6.3), 5.38 (5-H; dd, 15.3 / 7.7). 4.13 (3-H; bs), 2.45 (1-H; d, 1.5), 2.38 (4-H; m), 2.20 (OH; bd, 3.3), 1.68 (7-H; d, 6.2), 1.09 (4-CH ₃ ; d, 6.8), ¹³ C NMR (CDCl ₃ )

131.5 (5), 127.9 (6), 83.5 (2), 73.6 (1), 66.2 (3), 43.4 (4), 18.1 (7), 15.7 (4-Me).

 (3S, 4R) -3-tert-butyldimethylsilyloxy-4-methylhept-5E-enal (D).

 To a stirred solution of alcohol C (248 mg, 2 mmol) and imidazole (340 mg, 5 mmol) in 3 ml of dry DMF was added tert-butyldimethylsilyl chloride (452 mg, 3 mmol). After stirring the mixture overnight, 10 ml of 10% NaOH solution was added to break up the excess tert-butyldimethylsilyl chloride. The product is extracted into ether and the extract is washed successively with water, 0.5 N HCl, and water, dried and evaporated. Purification of the residue by silica gel chromatography with hexane afforded 457 mg of (3R, 4R) -3-tert-butyldimethylsilyloxy-4-methylhept-5E-en-1-one (96% yield).

¹ H-NMR (CDCl ₃ ) δ 5.50 (6-H; dq, 15.3 / 6.1), 5.38 (5-H; dd, 15.3 / 7.5). 4.16 (3-H; dd, 5.7 / 1.7), 2.37 (1-H; d, 1.7), 2.35 (4-H; m), 1.68 (7-H; d, 6.1), 1.07 (4-Me; d, 6.8), 0.90 (CMe ₃ ; s), 0.12 (SiMe; s), 0.09 (SiMe; s).

 Using the same procedure, the corresponding TBDPS derivative (3R, 4R) -3-tert-butyldiphenylsilyloxy-4-methylhept-5E-en-1-one was obtained in 92% yield.

19.4

18.0 (7), 14.7 (4-Me).[α] _D + 32.9 ^° (c 3.0, CHCl ₃ ). ¹ H NMR (CDCl ₃ ) δ 7.72 / 7.38 (2Ph-H ₅ ), 5.32 (6-H; m), 5.25 (5-H; dd, 16.2 / 7.3), 4.29 (3-H; dd, 5.2 / 2.0), 2.38 (4-H; d, 2.0), 2.64 (4-H; d, 2.0), 1.64 (7-H; d, 5.3), 1.11 (4-Me; d, 6.9), 1.06 (CMe ₃ ) . ¹³ C NMR (CDCl ₃ ) δ 136.1 / 135.9 / 133.6 /129.7/129.6/127.5/127.3 (Ph), 132.4 (5), 126.1 (6), 83.3 (2), 73.5 (1), 68.0 (3), 43.6 (4), 26.9

19.4

18.0 (7), 14.7 (4-Me).

2-Methylbutene (1.15 ml, 2M solution in THF, 2.3 mmol) was added to 1.1 ml of a solution of BF ₃ in THF (1M, 1.1 mmol) at -25 ^° C and the mixture was stirred in an ice bath for two hours. Then the temperature was lowered to -50 ^o C and immediately added a solution of TBS derivative (238 mg, 1 mmol) in 1 ml of THF. The cooling bath was removed and the reaction mixture was allowed to warm to room temperature and left at room temperature for one hour. Then a 2.2 M solution of KH ₂ PO ₄ / K ₂ NRA ₄ (4.8 ml) and 30% H ₂ O ₂ (0.8 ml) were added at 0 ^° C. After 1 hour, THF was evaporated and the residue was extracted into ether. The dry ether extract was evaporated, the residue was chromatographed on silica gel (1% EtOAc / Hexane) to give 194 mg of aldehyde D (76% yield).

¹ H NMR (CDCl ₃ ) δ 9.78 (1-H; t, 2.3), 5.46 (6-H; dq, 15.3 / 6.1), 5.34 (5-H; dd, 15.3 / 7.5), 4.13 (3-H ; m), 2.47 (2-H; m), 2.31 (4-H; m). 1.66 (7-H; brd, 6.1), 0.99 (4-Me; d, 6.8), 0.87 (CMe ₃ ; s), 0.07 (SiMe; s), 0.04 (SiMe; s).

19.6

18.3 (7), 14.9 (4-Me).The aldehyde tert-butyl diphenyl silyl ether derivative (TBDPS) was obtained in 83% yield, ¹ H-NMR (CDCl ₃ ) δ 9.52 (1-H; t, 2.4), 7.69 / 7.40 (2Ph-H ₅ ), 5.28 (6-H ; m), 5.22 (5.H; dd, 16.2 / 6.2), 4.19 (3-H; m), 2.42 (2-H; m). 2.29 (4-H; m), 1.60 (7-H; d, 5.4), 1.07 (CMe ₅ ), 1.02 (4-Me; d, 6.9). ¹³ C NMR (CDCl ₃ ) δ 202.0 (1), 136.1 / 133.6 / 133.3 / 130.2 / 129.7 / 127.7 / 127.6 (Ph), 132.3 (5), 126.2 (6), 72.8 (3), 47.6 (2), 42.2 (4), 27.1

19.6

18.3 (7), 14.9 (4-Me).

Methyl (5S, 6R) -5-tert-butyldimethylsilyloxy-6-methyl-7-oxonone-2E, 7E-dienoate (E)
To a stirred solution of aldehyde D (0.74 g, 2.9 mmol) and trimethylphosphonoacetate (632 mg, 3.5 mmol) in 5 ml THF, cooled to -78 ^° С, tetramethylguanidine (435 μl, 3.5 mmol) was added. After 30 minutes, the cooling bath was removed and the mixture was stirred for an additional four hours. The mixture was neutralized with 1 N HCl and the product was extracted into ether. Evaporation of the dried ether extract left a residue which was chromatographed on silica gel (5% EtOAc / Hexane) to give 0.814 g E (yield 96%). ¹ H-NMR (CDCl ₃ ) δ 6.93 (3-H; dt, 15.6 / 7.8), 5.62 (2-H; dd, 15.6 / 1.2). 5.37 (8-H, m), 5.37 (7-H, m), 3.71 (OCH ₃ , s), 3.61 (5-H, m), 2.29 (4-H ₂ , m), 2.22 (6-H , m) 1.66 (9-H ₃ ; brd, 6.1), 0.99 (6-Me; d, 6.8), 0.88 (CMe ₃ ; s), 0.03 (SiMe; s), 0.01 (SiMe; s).

19.4

18.1 (9), 14.7 (6-Me).The aldehyde tert-butyl diphenylsilyl ether derivative (TBDPS) was obtained in 90% yield, ¹ H-NMR (CDCl ₃ ) δ 7.68 / 7.38 (2Ph-H ₅ ), 6.75 (3-H; dt, 15.6 / 7.4), 5.62 (2 -H; d, 15.6), 5.34 (8-H, m), 5.29 (7-H, m), 3.70 (5-H, m), 3.68 (OCH ₃ , s), 2.28 (4-H ₂ , m), 2.20 (6-H, m), 1.62 (9-H ₃ ; d, 5.3), 1.08 (CMe ₅ ), 0.99 (6-Me; d, 6.9), ¹³ C NMR (CDCl ₃ ) δ 166.7 (1), 146.4 (3), 136.0 / 134.2 / 133.8 / 129.62 / 129.56 / 127.5 / 127.4 (Ph), 132.5 (7), 125.8 (8), 122.6 (2), 76.2 (5), 51.3 (OCH ₃ ), 41.7 (6), 36.8 (4), 27.0

19.4

18.1 (9), 14.7 (6-Me).

Methyl (5S, 6R) -5-tert-butyldimethylsilyloxy-6-methyl-7-oxohept-2 (E) -7-enoate (F)
Ozone was passed through a solution of methyl ester E (328 mg, 1.0 mmol) and 97 μl of pyridine in 15 ml of CH ₂ Cl ₂ at -78 ^° C and ozonolysis was monitored by thin layer chromatography. After the methyl ester was used up, about 500 mg of zinc dust and 1 ml of glacial acetic acid were added. The temperature was slowly raised to 25 ^° C. The mixture was filtered and the filtrate was washed successively with saturated solutions of CuSO ₄ and NaHCO ₃ . After evaporation of the solvent, the crude aldehyde F (249 mg, 83%) was used in the next step without further purification. ¹ H-NMR (CDCl ₃ ) δ 9.96 (7-H; t, 2.3), 6.96 (3-H, dt, 15.7 / 7.6), 5.90 (2-H, dd, 15.7 / 0.7), 4.05 (5- H; m), 3.74 (OMe; s), 2.51 (6-H; m), 2.45 (4-H ₃ ; m), 1.09 (6-Me; d, 6.9), 0.88 (CMe ₃ ; s), 0.04 (SiMe; s), 0.03 (SiMe; s).

Methyl (5S, 6R) -5-tert-butyldimethylsilyloxy-6-methyl-8-phenyl-octa-2 (E), 7E-dienoate (G)
To a stirred solution of aldehyde F (25.0 mg, 0.08 mmol) in 1.5 ml of THF at -78 ^° C was added 0.80 ml of a cooled (-78 ^° C) mixture of benzyltriphenylphosphonium chloride (268 mg, 0.69 mmol, in 6.9 ml of THF) and n- butyllithium (280 μl, 2.5 M in hexane). After 15 minutes, the cooling bath was removed and stirring was continued for two hours. The reaction was quenched with a saturated solution of ammonium chloride and THF was evaporated. The concentrate was extracted into hexane twice and the combined extracts were washed with brine, dried and evaporated. The remaining oil, a 5: 1 mixture of E and Z isomers, was dissolved in 1.5 ml of benzene containing thiophenol (0.02 M) and 1.1'-azobis (cyclohexanecarbonitrile) (VAZO, 0.006 M), and the mixture was refluxed for 5 hours. After cooling to room temperature, hexane (15 ml) was added and the organic solution was washed successively with 10% NaOH and brine, dried (MgSO ₄ ) and evaporated. Chromatography of the residue on silica gel (2% EtOAc / Hexane) gave 24 mg (80% yield) of product G.

18.1

16.2 (6-Me), -4.4 (Si-Me), -4.5 (Si-Me), Calcd for C₂₂H₃₄O₃Si: С, 70.52; Н, 9.17. Found: С. 70.72; Н, 9.42.[α] _D +68.2 ^° (c 1.5, CHCl ₃ ); EIMS m / z 374 (<1%; M ⁺ ), 359 (1; M ⁺ -CH ₃ ), 317 (10; M ⁺ -Bu), 275 (10), 243, (73), 143 (20) 115 (10), 97 (64), 89 (31), 73 (100); HREIMS m / z 374.2232 (C ₂₂ H ₃₄ O ₃ Si, Δ + 4.5 mmu), 359.2031 (C ₂₁ H ₃₁ O ₃ Si, Δ + 1.1 mmu). 317.1579 (C ₁₈ H ₂₅ O ₃ Si, Δ-0.6 mmu); UV (MeOH) λ _max (ε) 206 (33500), 252 (20100) nm; IR ν _max 2952, 2855, 1725, 1657, 1435, 1257, 1168, 1097, 970, 836, 775 cm ^-1 ; ¹ H NMR δ 7.2-7.4 (Ph-H ₅ ; m), 6.96 (3-H; ddd, 15.6 / 7.8 / 7.5), 6.37 (8-H; d, 15.9), 6.16 (7-H; dd, 15.9 / 8.1), 5.84 (2-H; d, 15.6), 3.75 (5-H; ddd, 10.2 / 6.0 / 4.2), 3.72 (OMe; s), 2.44 (6-H;
m), 2.36 (4-H ₂ ; m), 1.10 (6-Me; d, 6.9), 0.91 (Si-CMe ₃ ; s), 0.06 (Si-Me, s), 0.05 (Si-Me; s ); ¹³ C NMR δ 166.8 (1), 146.4 (3), 137.6 (Ph 1 '), 131.9 (8), 130.4 (7), 128.5 (Ph 3' / 5 '), 127.0 (Ph 4'), 126.0 ( Ph 2 '/ 6'), 122.9 (2), 75.0 (5), 51.4 (OMe), 42.8 (6), 37.6 (4), 25.9

18.1

16.2 (6-Me), -4.4 (Si-Me), -4.5 (Si-Me), Calcd for C ₂₂ H ₃₄ O ₃ Si: C, 70.52; H, 9.17. Found: S. 70.72; H, 9.42.

(5S, 6R) -5-tert-butyldimethylsilyloxy-6-methyl-8-phenyllocta-2E, 7E-dienoic acid (H)
To a solution of Cr ester (159 mg, 0.43 mmol) in 7 ml of acetone was added 5 ml of 1 N LiOH. The mixture was stirred at 25 ^{° C.} for 3 hours, diluted with 20 ml Et ₂ O and oxidized with 1 N Hcl to ~ pH 4. The organic layer was separated and washed with 20 ml of brine and water, dried (Mq SO ₄ ) and evaporated. The remaining oil was chromatographed on silica gel (40% EtOAc in hexane containing 0.5% AcOH to give pure acid H as a pale yellow, low viscosity oil (145 mg, 95% yield).

18.1

16.1 (6-Ме), -4.4 (Si-Me), -4.5 (Si-Me).[α] _D +87.0 ^° (c 1.4, CHCl ₃ ); EIMS m / z: 343 (1; M ⁺ -OH), 303 (5), 275 (9), 257 (4), 229 (62), 213 (16), 171 (22), 143 (37), 131 (16), 115 (23), 97 (100), 91 (44); HREIMS m / z 343.2107 (С ₂₁ Н ₃₁ O ₂ Si, Δ-1.3 mmu), 229.1220 (С ₁₅ Н ₁₇ O ₂ , Δ + 0.9 mmu); UV λ _max (ε) 206 (24500), 252 (15600) nm; IR ν _max 3300-2800 (br), 2956, 2856, 1697, 1651, 1419, 1256, 1097, 836, 693 cm ^-1 , ¹ H NMR δ 10.4 (CO ₂ H; bs, W _1/2 ≈100) 7.2-7.4 (Ph-H ₅ ; m), 7.09 (3-H; ddd, 15.6 / 7.6 / 7.6), 6.39 (8-H; d, 15.9), 6.16 (7-H; dd, 15.9 /8.1), 5.85 (2-Н; d, 15.6), 3.78 (5-Н; ddd, 6.0 / 6.0 / 4.2), 2.46 (6-Н; m), 2.40 (4-Н ₂ ; m), 1.12 (6-Me; d, 6.9), 0.92 (Si-CMe ₃ ; s), 0.07 (SiMe ₂ ; s); ¹³ C NMR δ 171.6 (1), 149.1 (3), 137.5 (Ph 1 '), 131.8 (8), 130.5 (7), 128.5 (Ph 3' / 5 '), 127.1 (Ph 4'), 126.1 ( Ph 2 '/ 6'), 122.7 (2), 74.9 (5), 42.9 (6), 37.6 (4), 25.8

18.1

16.1 (6-Me), -4.4 (Si-Me), -4.5 (Si-Me).

3- (3-Chloro-4-methoxyphenyl) -D-alanine 2,2,2-trichloroethyl ester (I)
A sample of the BOC D-chlorothyrosine derivative (160 mg, 0.35 mmol) was dissolved in 3 ml of pure trifluoroacetic acid and left at room temperature for 1 hour. Excess reagent was removed under reduced pressure, returning the desired amine I as trifluoroacetate salt (165 mg, 100% yield).

56.1 (OMe), 54.2 (2), 34.9 (3).[α] _D +1.7 ^o (c 5.2, CHCl ₃ ); IR ν _max 3400-2500 (br), 1760, 1680, 1500, 1200, 1130, 1070, 805, 710 cm ^-1 ; ¹ H NMR δ 8.07 (NH ₂ ; br m, W _1/2 ≈45), 7.27 (5-H; s), 7.12 (9-H; d, 8.1), 6.88 (8-H; d, 8.1) , 4.86 / 4.67 (CH ₂ Cl ₃ ; AB q, -12.0), 4.41 (2-Н; bs, W _1/2 ≈20), 3.86 (OMe; s), 3.33 (3-H; dd, -14.4 /3.6), 3.22 (3-H '; dd, -14.4 / 6.6); ¹³ C NMR δ 167.6 (1), 155.0 (7), 130.9 (5), 128.8 (9), 125.4 (4), 123.1 (6), 112.7 (8), 93.4 (CCl ₃ ), 75.3

56.1 (OMe), 54.2 (2), 34.9 (3).

18.1

16.5 (6-Ме), -4.3 (Si-Me), -4.6 (Si-Me); unit B δ 170.1 (1), 154.3 (7), 131.1 (5), 128.5 (4/9), 122.6 (6), 112.2 (8), 94.2 (CCl₃), 74.8

56.1 (OMe), 53.0 (2), 36.5 (3).Compound J
To a stirred solution of H (25 mg, 0.07 mmol) in 3 ml of anhydrous DMF in an argon atmosphere was added pentafordiphenylphosphinate (FDPP, 32 mg, 0.08 mmol), trifluoroacetate salt 1 (35 mg, 0.07 mmol) and diisopropylethylamine (DIEA, 27 mg, approximately 36 μl, 0.21 mmol, approximately 3 equivalents). Stirring was continued for one hour at 25 ^{° C.} and then the reaction mixture was extracted with 20 ml of Et ₂ O. The ether extract was washed with 10 ml of 1 N HCl, followed by washing with 10 ml of a saturated NaHCO ₃ solution, 20 ml of brine and 20 ml of water, dried ( MgSO ₄ ) and evaporated. The remaining pale yellow oil was chromatographed on silica gel (15% EtOAc in hexane) to give J as a colorless oil (32 mg, 65% yield):
[α] _D + 11.8 ^° (c 1.2, CHCl ₃ ); EIMS m / z: 644/646/648/650 (7/8/6/3; M ⁺ -Bu), 570/572/574 (46/100/21), 536/538 (18/15), 394 / 396 (67/29), 275 (20), 155/157 (29/9); HREIMS m / z 644.0981 (C ₂₉ H ₃₄ Cl ₄ NO ₅ Si, Δ-2.13 mmu): UV λ _max (ε) 204 (54900), 230 (23200), 248 (19200), 284 (3500) nm; IR ν _max 3290, 2980, 2850, 1760, 1680, 1640, 1505, 1380, 1270, 1169, 990, 720 cm ^-1 ; ¹ H NMR unit A δ 1.2-7.4 (Ph-H ₅ ; m), 6.87 (3-H; ddd, 15.0 / 7.8 / 7.5), 6.37 (8-H; d, 16.2), 6.18 (7-H; dd. 16.2 / 8.1), 5.82 (2-H; d, 15.0), 3.75 (5-H; ddd, 9.9 / 6.0 / 4.8), 2.46 (6-H; m), 2.36 (4-H ₂ ; m), 1.11 (6-Me; d, 6.9), 0.91 (SiCMe ₃ ; s), 0.07 (SiMe; s), 0.06 (SiMe; s); unit B δ 7.19 (5-H; d, 2.1), 7.04 (9-H; dd, 8.4 / 2.1), 6.85 (8-H; d, 8.4), 5.85 (NH, d, 7.8), 5.08 (2 -H; ddd, 7.8 / 6.0 / 5.7), 4.81 / 4.74 (CH ₂ CCl ₃ ; AB q, -11.7), 3.87 (OMe; s), 3.22 (3-H, dd, -14 1 / 5.7), 3.12 (3-H '; dd, -14.1 / 6.0), ¹³ C NMR unit A δ 165.1 (1), 143.0 (3), 137.6 (9), 132.0 (8), 130.4 (7), 128.5 (11 / 13), 127.0 (12), 126.0 (10/14), 124.7 (2), 75.0 (5), 42.6 (6), 37.6 (4), 25.9

18.1

16.5 (6-Me), -4.3 (Si-Me), -4.6 (Si-Me); unit B δ 170.1 (1), 154.3 (7), 131.1 (5), 128.5 (4/9), 122.6 (6), 112.2 (8), 94.2 (CCl ₃ ), 74.8

56.1 (OMe), 53.0 (2), 36.5 (3).

56.1 (OMe), 53.0 (2), 36.5 (3).(1 ', R, 5S, 6R) -N-1' - (carbo-2``, 2 '', 2 '' - trichloroethoxy) -2 '- (3-chloro-4-methoxyphenyl) ethyl-5- tert-butyldimethylsilyloxy-6-methyl-8-phenyl-octa-2E, 7E-dienamide (K)
To a solution of J (50 mg, 0.07 mmol) in 4 ml of MeCN was added 400 μl of a 50% aqueous HF solution and the mixture was stirred for 1 hour at 25 ^° C. Extraction in 30 ml of Et ₂ O followed by washing the ether extract with 30 ml portions of solution of NaHCO ₃ , brine and water, drying (MgSO ₄ ) and evaporation gave alcohol K in the form of a colorless foam (40 mg, 95% yield):
[α] _D -6.1 ^° (c 1.3, CHCl ₃ ); EIMS m / z (rel intensity) 587/589/591/593 (M ⁺ , <1%), 552/554/556 (1/2 / 0.5), 456/458/460/462 (1/2/1 /0.2), 342/344/346 (7/8/4), 212/214 (15/5), 195/197 (6/2), 155/157 (99/34), 131 (100), 91 (77); HREIMS m / z 587.0721 (C ₂₇ H ₂₉ ³⁵ Cl ₄ NO ₅ , Δ + 7.9 mmu); UV λ _max 204 (56500), 230 (22100), 248 (18100), 284 (3600) nm; IR ν _max 3400, 3300, 2980, 1780, 1680, 1640, 1505, 1270, 1180, 1090, 1000, 770 cm ^-1 , ¹ H NMR unit A δ 7.2-7.4 (Ph-H ₅ ; m), 6.92 ( 3-H; ddd, 15.3 / 7.8 / 7.5), 6.46 (8-H; d, 15.9), 6.14 (7-H; dd, 15.9 / 8.4), 5.90 (2-H; d, 15.3), 3.65 ( 5-H; ddd 7.8 / 5.6 / 4.0), 2.39 (6-H / 4-H ₂ ; bm), 1.78 (OH; bs, W _1/2 ≈40 Hz), 1.14 (6-Me; d, 6.9 ); unit B δ 7.18 (5-H; d, 1.8), 7.03 (9-H; dd, 8.4 / 1.8), 6.84 (8-H; d, 8.4), 5.97 (NH; d, 7.8), 5.06 (2 -H; ddd, 7.8 / 6.0 / 5.7), 4.79 / 4.72 (CH ₂ CCl ₃ ; AB q, -12.0), 3.86 (OMe; s), 3.20 (3-H; dd, -14.1 / 5.7) 3.10 (3-H '; dd, -14.1 / 6.0). ¹³ C NMR unit A δ 165.3 (1), 142.6 (3), 137.0 (9), 131.7 (8), 131.0 (7), 128.5 (11/13), 127.3 (12), 126.1 (10/14), 125.0 (2), 73.8 (5), 43.2 (6), 37.2 (4), 16.8 (6-Me); unit B δ 170.2 (1), 154.2 (7), 131.0 (5), 128.4 (9), 128.3 (4), 122.5 (6), 112.2 (8), 94.2 (CCl ₃ ), 74.7

56.1 (OMe), 53.0 (2), 36.5 (3).

 3- (tert-butoxycarbonyl) amino-2,2-dimethylpropanoic acid (M).

79.7

68.1 (1), 47.1 (3), 36.7 (2), 28.3

22,4 (2-Мe₂).To a solution of 3-amino-2,2-dimethylpropan-1-ol (L) (3 g, 29 mmol) in 51 ml of a 10% solution of triethanolamine in MeOH was added di-tert-butyl dicarbonate (6.7 g, 31 mmol) and the mixture stirred at 25 ^o C for 1 hour. After removing the solvent, the residue was dissolved in CH ₂ Cl ₂ (30 ml) and the solution was washed twice with a 1 M KHSO ₄ solution (pH 2) and once with a saturated NaCl solution and dried (MgSO ₄ ). Removal of the solvent in vacuo afforded 5.8 g, (93% yield) of 3- (tert-butoxycarbonyl) amino-2,2-dimethylpropanol as a white solid, which was directly used for the next step without further purification (above 95% purity by NMR analysis), mp 70.5-71.5 ^o C;
IR ν _max 3350, 1685, 1456 cm ^-1 ; ¹ H NMR δ 4.87 (NH; br s), 3.72 (OH; br s), 3.19 (1-H ₂ ; d, 5.1), 2.95 (3-H ₂ , d, 6.0), 1.44 (CMe ₃ ; s ), 0.85 (2-Me ₂ ; s); ¹³ C NMR (CDCl ₃ ) δ 157.6

79.7

68.1 (1), 47.1 (3), 36.7 (2), 28.3

22.4 (2-Me ₂ ).

79.6

49.5/47.9 (2/3), 28.4

22.9 (2-Ме₂).To a solution of alcohol 3- (tert-butoxycarbonyl) amino-2,2-dimethylpropanol (5.3 g, 25.9 mmol) and sodium periodate (16.6 g, 77.7 mmol) in carbon tetrachloride (52 ml), acetonitrile (52 ml) and water ( 78 ml) ruthenium trichloride hydrate (122 mg) was added and the mixture was stirred at 25 ^{° C.} for 1 hour. The mixture was filtered through celite and a saturated solution of potassium carbonate in water (50 ml) was added. The aqueous layer was separated, washed with ether (20 ml), acidified with HCl to pH 2 at 0 ^{° C.} and extracted with CH ₂ Cl ₂ (and extracted 3 times with 30 ml). The combined extracts were washed with saturated NaCl and dried (MgSO ₄ ). Removing the solvent in vacuo gave a residue, which was first subjected to flash chromatography with reverse phase on a C18 silicon column (ODS 120 A, 50 to 90% MeOH) and then crystallized from ether to obtain 3.7 g, (yield 66%) M as white solid, mp 106-108 ^o C;
EIMS m / z (rel intensity) 217 (0.1), 161 (11), 98 (25), 88 (71), 57 (100); HREIMS m / z, 217.1292 (C ₁₀ H ₁₉ NO ₄ , Δ + 2.2 mmu); IR ν _{max /} 3450-2500, 1710, 1694, 1510 cm ^-1 ; ¹ H NMR of major conformer δ 5.03 (NH; br s), 3.26 (3-H ₂ ; m), 1.45 (CMe ₃ ; s), 1.24 (2-Me ₂ ; s); ¹³ С NMR (СОСl ₃ ) δ 183.2 (1), 156.3

79.6

49.5 / 47.9 (2/3), 28.4

22.9 (2-Me ₂ ).

 Allyl (2S) -2-hydroxy-4-methylpentanoate (N).

To a solution of 2.66 g of L-leucinic acid (20 mmol) and 1.74 g of sodium bicarbonate (20 mmol) in 30 ml of water at 0 ^° C were added 30 ml of a solution of 6.44 g of tetrabutylammonium chloride (20 mmol) and 1.74 ml of allyl bromide (20 mmol) ) in CH ₂ Cl ₂ . After vigorously stirring the mixture for 24 hours, CH ₂ Cl _{2 was} evaporated. About 50 ml of water was added and the aqueous layer was extracted four times with Et ₂ O. The ethereal solution was dried over anhydrous sodium sulfate and then evaporated. The residue was passed through a short Si column to obtain 3.21 g of allyl ether N (yield 93%) as a colorless oil,
[α] _D -8.4 ^° (c 1.1, CHCl ₃ ); IR ν _max 3464, 2957, 1732, 1203, 1140, 1087 cm ¹ ; ¹ H NMR δ 5.92 (allyl 2-H; m), 5.34 (allyl 3-H _Z ; dd, 17.4 / 1.1), 5.28 (allyl 3-H _E ; dd, 10.5 / 1.1), 4.67 (allyl 1-H ₂ ; d, 5.7), 4.23 (2-H; br s), 2.64 (OH; br s), 1.89 (4-H; m), 1.57 (3-H ₂ ; m), 0.96 (5-H ₃ ; d, 6.5), 0.95 (4-Me; d, 6.7); ¹³ C NMR δ 175.3 (1), 131.4 (allyl C-2), 118.6 (3), 68.9 (2), 65.7 (allyl C-1), 43.2 (3), 24.1 (4), 23.0 (5), 21.3 (4-Me).

 Allyl (2S) -2- [3 '(tert-butoxycarbonyl) amino-2', 2'-dimethylpropanoyloxy] -4-methylpentanoate (O).

79.0

48.6 (3), 44.0 (2), 28.4

22.2/23.0 (2-Me₂); unit D δ 170.6 (1), 131.4 (allyl C-2), 119.1 (allyl C-3), 70.9 (2), 66.0 (allyl C-1), 39.5 (3), 24.8 (4), 23.0 (5), 21.5 (4-Ме).To a solution of 0.8 g of M (3.7 mmol), 0.76 g of N (4.4 mmol) and 92 mg of DMAP in 10 ml of dry CH ₂ Cl ₂ at 0 ^° С, 0.84 g of DDC (4.1 mmol) in CH ₂ Cl _{2 was} added. The mixture was stirred at 25 ^{° C.} for 3 hours and filtered. The filtrate was washed with saturated aqueous sodium bicarbonate, dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (silica gel, 5% EtOAc / Hexane) gave 1.0 g (92% yield) of pure O as a colorless oil, R _f 0.68 (17:83 EtOAc / Hexane),
[α] _D -29.4 ^° (c 18.1, CHCl ₃ ); EIMS m / z (rel intensity) 371 (2, M ⁺ ), 242 (13), 184 (12), 126 (20), 84 (100); HREIMS m / z 371.2317 (C ₁₉ H ₃₃ NO ₆ , Δ-0.9mmu); IR (neat) ν _max 3385, 2963, 1731, 1720, 1513 cm ^-1 ; ¹ H NMR (300 MHz, CDCl ₃ ) unit C δ 5.39 (NH; obscured br s), 3.33 (3-H; dd, -13.5 / 7.4), 3.27 (3-H '; dd, -13.5 / 5.9), 2.78 (2-H, m). 1.44 (CMe ₃ ; s), 1.23 (2-Me; s), 1.22 (2-Me; s); unit D δ 5.91 (allyl 2-H, ddt, 16.6 / 10.3 / 6.0 Hz), 5.34 (allyl 3-H ₂ ; bd, 16.6), 5.27 (allyl 3-H _E ; bd, 10.3), 5.08 ( 2-H; dd, 9.6 / 3.6), 4.65 (allyl 1-H ₂ ; m), 1.6-1.9 (3-H ₂ /4-H; m), 0.94 (5-H ₃ ; d, 6.3), 0.94 (4-Me; d, 7.3). ¹³ C NMR unit C δ 176.5 (1), 156.3

79.0

48.6 (3), 44.0 (2), 28.4

22.2 / 23.0 (2-Me ₂ ); unit D δ 170.6 (1), 131.4 (allyl C-2), 119.1 (allyl C-3), 70.9 (2), 66.0 (allyl C-1), 39.5 (3), 24.8 (4), 23.0 (5 ), 21.5 (4-Me).

 (2S) -2- [3 '(tert-butoxycarbonyl) amino-2', 2'-dimethylpropanoyloxy] -4-methylpentanoic acid (P).

br s), 1.22 (2-Me; s), 1.21 (2-Me; s); unit D δ 6.47 (1-ОН; br s, W_1/2≈35), 5.09 (2-H; dd, 9.3/2.7), 1.7-1.9 (3-Н₂/4-Н; m), 0.97 (5-H₃; d, 6.0), 0.94 (4-Me; d, 6.0). ¹³С NMR (СDСl₃) unit С δ 176.5 (1), 156.5

79.3

48.6 (3), 44.0 (2), 28.3

23.0 (2-Me), 22.2 (2-Me); unit D δ 175.4 (1), 70.6 (2), 39.5 (3), 24.8 (4), 23.0 (5), 21.4 (4-Me).To 10 ml of a solution of 180 mg (0.49 mmol) of 0 and 60 mg (0.05 mmol) of tetrakis (triphenylphosphine) palladium in dry THF (in an argon atmosphere), 470 μl (5.4 mmol) of dry morpholine was slowly added over 10 minutes. After stirring for 50 minutes, 40 ml of ether was added and the solution was washed with 1 N HCl (40 ml) and then extracted with saturated sodium bicarbonate solution (2 times 30 ml). The aqueous extract was acidified with a solution of 0.5 N HCl and extracted with ether (40 ml). The ether extract was washed with water (2 times, 30 ml), dried (MgSO ₄ ) and evaporated in vacuo to give P as a colorless mobile oil (152 mg, 95% yield):
[α] _D -22.2 ^° (c 2.2, CHCl ₃ ); EIMS m / z (rel intensity) 331 (1, M ⁺ ), 275 (1), 258 (4), 231 (9), 202 (36), 174 (13), 144 (31), 126 (16) 114 (14), 98 (54), 88 (50), 84 9100); HREIMS m / z 331.2004 (C ₁₆ H ₂₉ NO ₆ , Δ-1.0 mmu). ¹ H NMR (CDCl ₃ ) unit C δ 5.41 (NH; dd, 5.7 / 5.4), 3.30 (3-H ₂ ; m), 2.68 (2-H; m), 1.43

br s), 1.22 (2-Me; s), 1.21 (2-Me; s); unit D δ 6.47 (1-OH; br s, W _1/2 ≈35), 5.09 (2-H; dd, 9.3 / 2.7), 1.7-1.9 (3-Н ₂ /4-Н; m), 0.97 (5-H ₃ ; d, 6.0), 0.94 (4-Me; d, 6.0). ¹³ C NMR (CDCl ₃ ) unit C δ 176.5 (1), 156.5

79.3

48.6 (3), 44.0 (2), 28.3

23.0 (2-Me); 22.2 (2-Me); unit D δ 175.4 (1), 70.6 (2), 39.5 (3), 24.8 (4), 23.0 (5), 21.4 (4-Me).

 Compound O.

74.6

56.1 (7-ОМe), 53.2 (2), 36.6 (3); unit С 176.9 (1), 156.4

79.1

48.7 (3), 44.0 (2), 22.8 (2-Ме), 22.3 (2-Me'); unit D 170.7 (1), 71.4 (2), 39.5 (3), 28.4

24.8 (4), 23.0 (4-Ме), 21.4 (5),
Аминокислота R.To a solution of alcohol K (80mg, 0.14 mmol), acid P (68mg, 0.21 mmol) and DMAP (4 ml) in dry CH ₂ Cl ₂ (4 mL) stirring at 0 ^o C under argon, was added DDC ( 44 mg, 0.21 mmol) in dry CH ₂ Cl ₂ (1 ml). The mixture was stirred at 0 ^{° C.} for 30 minutes, during which a white precipitate formed, and then allowed to warm to room temperature and was stirred for an additional 4 hours. The precipitate was filtered off and the filtrate was diluted with Et ₂ O (40 ml) and washed successively with dilute HCl (1 M, 40 ml), saturated aqueous sodium bicarbonate (40 ml), and brine (40 ml). The ether layer was dried (Mg ₂ SO ₄ ) and evaporated in vacuo to give a waxy solid. Chromatography (silica gel, EtOAc: 1: 3 hexane) gives pure Q as a colorless viscous oil (103 mg, 84% yield),
[α] _D -3.1 ^o (c 2.9. CHCl ₃ ); EIMS m / z 800/802/804/806 (<1, M ⁺ -C ₅ H ₈ O ₂ ), 415/417/419/421 (5/3/3/2), 342/344/346 (7 / 9/4), 286/288/290 (2/6/2), 207 (34), 178 (22), 155/157 (66/24), 131 (36), 91 (70), 70 ( 100); HREIMS m / z 800.2179 (C ₃₈ H ₄₈ N ₂ O ₈ ³⁵ Cl ₄ , Δ-1.4 mmu); UV (MeOH) λ _max (ε) 204 (51200), 230 (18500), 248 (17200), 282 (2200) nm; IR (NaCl) ν _max 3376, 2965, 1755, 1728, 1712, 1678, 1504, 1258, 1150, 1067, 732 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A: 7.28-7.33 (10-Н / 14-Н / 11-Н / 13-Н; m), 7.22 (12-Н; m), 6.78 (3-Н; ddd , 15.8 / 6.4 / 6.3), 6.40 (8-H; d, 15.8), 6.01 (7-H; dd, 15.8 / 8.7), 5.88 (2-H; d, 15.8), 5.06 (5-H; bm , W _1/2 ≈20 Hz), 2.62 (6-Н; M), 2.53 (4-Н ₂ , bm, W _1/2 ≈15 Hz), 1.12 (6-СН ₃ ; d, 6.8); unit B 7.18 (5-H; d, 2.0), 7.05 (9-H; dd, 8.5 / 2.0), 6.83 (8-H; d, 8.5), 6.49 (NH; d, 7.9), 5.06 (2- H; bm, W _1/2 ≈20 Hz), 4.79 / 4.70 (CH ₂ CCl ₃ ; ABq, -11.7), 3.85 (OCH ₃ ; s), 3.20 (3-Н _b ; dd, -14.1 / 5.8) 3.07 (3-H ₂ ; dd, -14.1 / 6.7); unit C 5.38 (NH; bt, 6.5), 3.27 (3-H ₂ ; d, 6.5), 1.20 (2-CH ₃ ; s), 1.15 (2-CH ₃ ';s); unit D 4.92 (2-H; dd, 10.0 / 3.8), 1.72 (4-H; bm, W _1/2 ≈20 Hz), 1.67 (3-H _b ; ddd, -14.1 / 10.0 / 5.0 /), 1.56 (3-H ₂ ; ddd, -14.1 / 9.1 / 3.8), 1.43 (СО ₂ СМe ₃ ; s), 0.86 (4-СН ₃ ; d, 6.4), 0.82 (5-Н ₃ ; d, 6.4). ¹³ C NMR (CDCl ₃ ) δ unit A 165.4 (1), 139.3 (3), 136.9 (9), 131.7 (8), 130.1 (7), 128.6 (11/13), 127.5 (12), 126.2 (10 / 14), 125.4 (2), 76.5 (5), 41.1 (6), 33.4 (4), 16.7 (6-Me); unit B 170.0 (1), 154.1 (7), 131.2 (5), 128.8 (4), 128.5 (9), 122.3 (6), 112.1 (8), 94.3

74.6

56.1 (7-OMe), 53.2 (2), 36.6 (3); unit C 176.9 (1), 156.4

79.1

48.7 (3), 44.0 (2), 22.8 (2-Me), 22.3 (2-Me '); unit D 170.7 (1), 71.4 (2), 39.5 (3), 28.4

24.8 (4), 23.0 (4-Me), 21.4 (5),
Amino acid R.

Activated zinc dust (400 mg, excess) and AcOH (4 ml) were added to amino acid Q (100 mg, 0.11 mmol). The heterogeneous mixture was sonicated for 45 minutes, mixed for an additional 90 minutes at room temperature, and then poured onto a celite pad. Organic matter was washed off with a celite pad CH ₂ Cl ₂ . The solvent was removed in vacuo leaving the carboxylic acid as a colorless amorphous solid.

The resulting crude acid was dissolved without purification in trifluoroacetic acid (TFA, 5 ml) and allowed to remain at room temperature for 1 hour. After this time, the excess TFA was removed in vacuo and the resulting amorphous solid was then chromatographed (Sep-Pak ^™ , silica gel, first CH ₂ Cl ₂ , then 10% MeOH / CH ₂ Cl ₂ ) to give the desired compound as the trifluoroacetate ammonium salt . Repeated lyophilization of the aqueous salt solution gives the free amino acid R as a colorless amorphous solid (68 mg, 91% yield in two steps); IR (NaCl) ν _max 3300, 3200, 2965, 1693, 1606, 1504, 1441, 1259, 1201, 1146, 1066, 727 cm ^-1 . ¹ H NMR (CD ₃ OD) δ unit A: 7.33 (10-H / 14-H; d, 7.4), 7.28 (11-H / 13-H; t, 7.4), 7.18-7.23 (12-H; m), 6.69 (3-H; ddd, 15.6 / 7.7 / 7.0), 6.43 (8-H; d, 15.8), 6.04 (7-H; dd, 15.8 / 8.9), 6.00 (2-H; d, 15.6), 5.01 (5-Н; ddd, 9.1 / 6.9 / 3.1), 2.64 (4-Н _b ; bm, W _1/2 ≈30 Hz), 2.60 (6-Н; bm, W _1/2 ≈20 Hz), 2.49 (4-H ₂ ; ddd, 15.8 / 9.1 / 7.7), 1.13 (6-Me; d, 6.7); unit B 7.18-7.23 (5-H; m), 7.11 (9-H; dd. 8.3 / 1.6), 6.92 (8-H; d, 8.3), 4.59 (2-H; bm, W _1/2 ≈ 20 Hz). 3.81 (OCH ₃ ; s), 3.14 (3-H _b ; dd, -13.7 / 4.3), 2.96 (3-H ₂ ; m, W _1/2 ≈20 Hz); unit C 2.96 (3-H ₂ ; bm, W _1/2 ≈20 Hz), 1.31 (2-CH ₃ ; s), 1.25 (2-CH ₃ ';s); unit D 4.90 (2-H; dd, 9.6 / 4.0), 1.66 (4-H; bm, W _1/2 ≈25 Hz), 1.59 (3-H _b ; ddd, -14.4 / 9.6 / 4.8), 1.53 (3-H ₂ , ddd, -14.4 / 9.1 / 4.0), 0.81 (4-Me; d, 6.5), 0.74 (5-H ₃ ; d, 6.5). ¹³ C NMR (CD ₃ OD) δ unit A 167.7 (1), 140.7 (3), 138.4 (9), 133.0 (8), 131.7 (7), 129.6 (11/13), 128.5 (12), 127.3 ( 10/14), 127.1 (2), 78.4 (5), 43.1 (6), 35.7 (4), 17.4 (6-Me), unit B 179.8 (1), 155.2 (7), 132.3 (4), 132.1 (5), 130.1 (9), 123.0 (6), 113.4 (8), 56.6 (7-OMe), 56.6 (2), 37.8 (3), unit C 176.8 (1), 48.2 (3), 42.2 (2), 23.3 (2-Me), 23.3 (2-Me '), unit D 172.0 (1), 73.4 (2), 40.7 (3), 26.0 (4), 23.1 (4-Me), 21.8 (5).

Cryptophycin 51
To a stirred solution of amino acid T (75 mg, 0.11 mmol) in anhydrous DMF (20 ml) at room temperature in an argon atmosphere was added diisopropylethylamine (DIEA, 44 mg, 60 μl, 0.34 mmol, approximately 3 equivalents) followed by pentafluorodiphenylphosphinate (FDPP, 55 mg, 0.14 mmol, approximately 1.3 equivalents) in DMF (2 ml). The mixture was stirred for 12 hours, Et ₂ O (40 ml) and H ₂ O (40 ml) were added, dried (MgSO ₄ ) and evaporated under reduced pressure. The residual waxy solid was further purified by reverse phase chromatography (ODC, 10 μl 30% H ₂ O / MeCN, 3 ml min ^-1 ) to obtain Cryptophycin 51 as a colorless amorphous solid (45 mg, 61% yield),
[α] _D +26.4 ^° (c 0.25, CHCl ₃ ); EIMS m / z 652/654 (M ⁺ , 3/1), 632/634 (3/2), 426/428 (51/15), 227 (64), 195/197 (64/22), 155 / 157 (71/15), 131 (59), 91 (100); HREIMS m / z 652.2936 (C ₃₆ H ₄₅ N ₂ O ₇ ³⁵ Cl, Δ-2.1 mmu); UV (MeOH) λ _max (ε) 204 (52000), 228 (20400), 250 (13400), 284 (2800) nm; IR (NaCl) ν _max 3376, 3270, 2960, 1747, 1721, 1659, 1536, 1514, 1259, 1150, 1066, 1013, 980, 694 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A 7.32 (10-H / 14-H; dd. 8.0 / 1.5), 7.29 (11-H / 13-H; t, 8.0), 7.24 (12-H; bm, W _1/2 ≈15 Hz), 6.77 (3-Н; ddd, 15.2 / 10.8 / 4.3), 6.40 (8-Н; d, 15.8), 6.01 (7-Н; dd, 15.8 / 8.8), 5.76 ( 2-Н; dd, 15.2 / 1.1), 5.04 (5-Н; ddd, 11.1 / 6.4 / 1.9), 2.54 (4-Н _b / 6-Н; bm, W _1/2 ≈15 Hz), 2.37 ( 4-H _a ; ddd, -14.3 / 11.1 / 10.8), 1.13 (6-Me; d, 6.8); unit B 7.20 (5-H; d, 2.0), 7.05 (9-H; dd, 8.4 / 2.0), 6.84 (8-H; d, 8.4), 5.61 (NH; d, 7.8), 4.74 (2- H; ddd, 7.8 / 7.6 / 5.4), 3.87 (OMe; s), 3.11 (3-H _b ; dd, -14.2 / 5.4), 3.06 (3-H _a ; dd, -14.2 / 7.6); unit С 7.24 (NH; bm. W _1/2 ≈15 Hz), 3.40 (3-H _b ; dd, -13.5 / 8.5), 3.12 (3-Н _a ; dd. -13.5 / 3.6), 1.22 (2 -Me; s), 1.15 (2-Me ', s); unit D 4.85 (2-H; dd, 10.2 / 3.6), 1.66 (3-H _b , ddd, -14.0 / 10.2 / 4.6), 1.61 (4-H; bm W _1/2 ≈20.0 Hz), 1.33 ( 3-H _a , ddd, -14.0 / 9.0 / 3.6), 0.74 (4-Me; d, 6.6), 0.72 (5-H ₃ ; d, 6.6). ¹³ C NMR (CDC1 ₃ ) δ unit A 165.1 (1), 142.2 (3), 136.7 (9), 131.7 (8), 130.1 (7), 128.6 (11/13), 127.5 (12), 126.1 (10/14), 124.6 (2), 77.0 (5), 42.2 (6), 36.5 (4), 17.3 (6-Me); unit B 170.3 (1), 154.1 (7), 130.9 (5), 129.5 (4), 128.3 (9), 122.5 (6), 112.3 (8), 56.1 (7-OMe), 54.2 (2), 35.3 (3); unit C 178.0 (1), 46.5 (3), 42.7 (2), 22.8 (2-Me), 22.6 (2-Me '), unit D 170.6 (1), 71.5 (2), 39.5 (3), 24.5 (4), 22.7 (4-Me), 21.2 (5).

 Example 9. Synthesis of Cryptophycin 52 and Cryptophycin 53.

To a stirred solution of Cryptophycin 51 (75 mg, 0.12 mmol) in anhydrous dichloromethane at 0 ^° C in an argon atmosphere was added a solution of m-chloroperbenzoic acid (mCPBA, 50 mg, 0.23 mmol, approximately 2 equivalents of 80% active oxygen) in dichloromethane (1 ml). After 30 minutes, the reaction mixture was allowed to warm to room temperature and stirred for an additional 12 hours. Then, the solvent was removed under reduced pressure to obtain a 1.8: 1 mixture of Cryptophycin 52 and 53 (according to NMR analysis), respectively, as an amorphous solid. A mixture of regioisomeric epoxides was dissolved in a minimum amount of acetonitrile and subjected to reverse phase chromatography (YMC-ODS, 10 μm, 250 mm • 22.5 mm, 30% H ₂ O / MeCN, 6 ml min ^-1 ) to separate Cryptophycin 52 (37 mg, yield 48%) and Cryptophycin 53 (19 mg, yield 25%).

Spectral data for Cryptophycin 52
[α] _D +19.9 ^° (c 0.5. CHCl ₃ ); EIMS m / z 668/670 (4/2, M ⁺ ), 445 (35), 244 (12), 227 (22), 195/197 (66/27), 184 (45), 155/157 (38 / 10), 91 (100); HREIMS m / z 668.2873 (C ₃₆ H ₄₅ N ₂ O ₈ ³⁵ Cl, Δ-0.9 mmu), 445.2497 (C ₂₅ H ₃₅ NO ₆ , Δ-3.3 mmu); UV (MeOH) λ _max (ε) 204 (35100), 218 (20900) nm; IR (NaCl) ν _max 3415, 3270, 2960, 1748, 1721, 1650, 1536, 1504, 1260, 1192, 1150, 1066, 1013, 800, 698 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A 7.33-7.38 (11-Н / 12-Н / 13-Н; bm, W _1/2 ≈25 Hz), 7.24 (10-H / 14-H; m, W _1/2 ≈15 Hz), 6.76 (3-H; ddd, 15.1 / 10.8 / 4.3), 5.71 (2-H; dd, 15.1 / 1.7), 5.20 (5-H; ddd, 11.0 / 5.0 / 1.8) , 3.68 (8-H; d, 1.9), 2.92 (7-H; dd, 7.5 / 1.9), 2.57 (4-H _b ; ddd, -14.6 / 1.8 / 1.7), 2.45 (4-H _a ; ddd , -14.6 / 11.0 / 10.8), 1.78 (6-H; bm, W _1/2 ≈15 Hz), 1.14 (6-Me; d, 6.9), unit B 7.18 (5-H; d, 2.2 ), 7.04 (9-H; dd, 8.4 / 2.2), 6.83 (8-H; d, 8.4), 5.56 (NH; d, 7.9), 4.73 (2-H; ddd, 7.9 / 7.4 / 5.3), 3.87 (OMe; s), 3.09 (3-H _b ; dd, -14.6 / 5.3), 3.05 (3-H _a , dd, -14.6 / 7.4); unit C 7.20 (NH; dd, 8.6 / 3.2), 3.41 (3-H _b ; dd, -13.4 / 8.6), 3.10 (3-H _a , dd, -13.4 / 3.2), 1.22 (2-Me; s ), 1.15 (2-Me ';s); unit D 4.82 (2-H; dd, 10.2 / 3.5), 1.73 (3-H _b ; bm, W _1/2 ≈20 Hz), 1.66 (4-H; bm, W _1/2 ≈20 Hz), 1.31 (3-H _a , ddd, -13.8 / 9.1 / 3.5), 0.84 (4-Me; d, 6.6), 0.82 (5-H ₃ ; d, 6.6); ¹³ C NMR (CDCl ₃ ) δ unit A 164.9 (1), 141.8 (3), 136.7 (9), 128.7 (11/13), 128.3 (12), 125.6 (10/14), 124.7 (2), 75.9 (5), 63.0 (7), 59.0 (8), 40.7 (6), 36.9 (4), 13.5 (6-Me), unit B 170.3 (1), 154.1 (7), 130.9 (5), 129.5 ( 4), 128.5 (9), 122.6 (6), 112.4 (8), 56.1 (7-OMe), 54.3 (2), 35.3 (3), unit С 178.0 (1), 46.5 (3), 42.8 (2 ), 22.8 (2-Me), 22.8 (2-Me '), unit D 170.5 (1), 71.2 (2), 39.3 (3), 24.6 (4), 22.7 (4-Me), 21.2 (5) .

Spectral data for Cryptophycin 53
[α] _D + 20.8 ^° (c 1.7, CHCl ₃ ); EIMS m / z 668/670 (5/4, M ⁺ ), 445 (32), 244 (15), 227 (24), 195/197 (64/21), 184 (60), 155/157 (33 / 9), 91 (100); HREIMS unit 668.2853 (C ₃₆ H ₄₅ N ₂ O _a ³⁵ Cl, Δ1.1 mmu); UV (MeOH) λ _max (ε) 204 (38700), 218 (22900) nm; IR (NaCl) ν _max 3415, 3280, 2917, 2849, 1748, 1722, 1660, 1504, 1465, 1260, 1190, 1150, 1066, 755 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A 7.29-7.36 (11-H / 12-H / 13-H, bm, W _1/2 ≈20 Hz), 7.23 (10-H / 14-H; dd, 8.3 /1.7), 6.77 (3-H; ddd, 15.1 / 10.9 / 4.3), 5.81 (2-H; dd, 15.1 / 1.3), 5.17 (5-H; ddd, 11.2 / 4.9 / 1.8), 3.58 (8-H; d, 1.7), 2.90 (7-H; dd, 7.8 / 1.7), 2.67 (4-H _b ; ddd, 14.7 / 11.2 / 10.9), 2.56 (4-H _a ; dddd, 14.7 / 4.3 / 1.8 / 1.3), 1.67-1.78 (6-H; bm, W _1/2 ≈45), 1.03 (6-CH ₃ ; d, 7.1); unit B 7.21 (5-H; d, 2.1), 7.07 (9-H; dd, 8.5 / 2.1), 6.84 (8-H; d, 8.4), 5.90 (2-NH; d, 7.9), 4.75 ( 2-H; ddd, 7.9 / 7.9 / 4.9), 3.85 (7-OCH ₃ ; s), 3.14 (3-H _b ; dd, 14.5 / 4.9), 3.03 (3-H _a ; dd, 14.5 / 7.9) ; unit C 7.29-7.36 (3-NH; bm, W _1/2 ≈25), 3.43 (3-H _b ; dd. 13.7 / 8.8), 3.10 (3-H _a ; dd, 13.7 / 3.4), 1.23 ( 2-CH ₃ ; s), 1.17 (2-CH ₃ ';s); unit D 4.92 (2-H; dd, 10.3 / 3.2), 1.67-1.78 (3-Н _b / 4-Н; bm, W _1/2 ≈45), 1.48 (3-H _a ; ddd, 13.9 / 8.8 / 3.2), 0.89 (4-CH ₃ ; d, 6.6), 0.86 (5-H ₃ ; d, 6.6). ¹³ C NMR (CDCl ₃ ) δ unit A 165.1 (1), 142.0 (3), 137.0 (9), 128.5 (11/13), 128.5 (12), 125.3 (10/14), 124.6 (2), 76.7 (5), 63.2 (7), 56.2 (8), 40.8 (6), 36.7 (4), 13.4 (6-Me); unit B 170.4 (1), 154.0 (7), 130.8 (5), 129.7 (4), 128.2 (9), 122.5 (6), 112.3 (8), 56.1 (7-OMe), 54.4 (2), 35.3 (3); unit C 177.9 (1), 46.4 (3), 42.7 (2), 23.0 (2-Me), 22.7 (2-Me '); unit D 170.5 (1), 71.3 (2), 39.2 (3), 24.7 (4), 22.8 (4-Me), 21.3 (5).

Example 10. Synthesis of Cryptophycin 55
To a solution of Cryptophycin 52 (6 mg, 0.009 mmol) in 0.6 ml of a 2: 1 mixture of 1,2-dimethoxyethane / water was added 2 μl of a 12 N HCl solution. The solution was allowed to stir at room temperature for 20 hours, neutralized with potassium carbonate, filtered through a 5 micron filter and evaporated. Acetonitrile insoluble matter was purified by reverse phase gel permeation chromatography on a C18 column (250 x 10 mm column) using a 4: 1 MeOH / H ₂ O mixture to give 3.0 mg Cryptophycin 55 (48% yield) .

[α] _D +42.5 ^° (c 1.1, CHCl ₃ ); EIMS m / z 704/706/708 (M ⁺ <1), 668/670 (1.5 / 0.5, M ⁺ -HCl), 445 (6), 226 (8), 195/197 (16/5), 184 (10), 155/157 (33/11), 135 (100), 91 (99), 77 (30); HREIMS m / z 668.2873 (M ⁺ -HCl, C ₃₆ H ₄₅ N ₂ O ₈ ³⁵ Cl, Δ-0.8 mmu); UV (MeOH) λ _max (ε) 204 (48400), 218 (29200), 284 (1600) am; IR (NaCl) ν _max 3410, 3286, 2959, 1748, 1723, 1666, 1538, 1504, 1455, 1257, 1178, 1066, 753 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A 7.35-7.42 (10-Н / 11-Н / 12-Н / 13-Н / 14-Н; m), 6.78 (3-H; ddd, 15.1 / 10.6 / 4.5 ), 5.78 (2-H; dd, 15.1 / 1.7), 5.16 (5-H; ddd, 11.1 / 8.3 / 2.1), 4.65 (8-H; d, 9.7), 4.01 (7-H; bd, 9.7 ), 2.69 (4-H _b ; dddd, -14.5 / 4.5 / 2.1 / 1.7), 2.50 (6-H; bm, W _1/2 ≈15), 2.38 (4-H _a ; ddd, -14.5 / 11.1 / 10.6), 1.53 (7-OH, s), 1.04 (6-Me, d, 7.1); unit B 7.21 (5-H; d, 2.2), 7.07 (9-H; dd, 8.5 / 2.2), 6.85 (8-H; d. 8.5), 5.57 (2-NH; d, 7.8), 4.74 ( 2-H; ddd, 7.8 / 7.6 / 5.2), 3.88 (7-OCH ₃ ; s), 3.13 (3-H _b ; dd. 14.5 / 5.2), 3.05 (3-H _a ; dd, 14.5 / 7.6) ; unit C 7.21 (3-NH; m), 3.38 (3-H _b ; dd, 13.5 / 8.3), 3.17 (3-H _a ; dd, 13.5 / 4.1), 1.23 (2-CH ₃ ; s), 1.17 (2-CH ₃ ; s), unit D 4.93 (2-H; dd, 10.1 / 3.5), 1.78 (3-H _b ; ddd, 13.5 / 10.1 / 5.0), 1.72 (4-H; bm, W _{1 / 2} ≈20), 1.43 (3-H _a ; ddd, 13.5 / 8.8 / 3.5), 0.92 (4-CH ₃ ; d. 6.6), 0.92 (5-H ₃ , d, 6.4), ¹³ C NMR ( CDCl ₃ ) δ unit A 165.1 (C-1), 142.4 (C-3), 138.4 (C-9), 129.0 (C-11/13), 128.3 (C-12), 128.0 (C-10/14 ), 124.6 (С-2), 76.1 (С-5), 74.1 (С-7), 62.0 (С-8), 38.4 (С-6), 36.5 (С-4), 8.6 (6-Me) ; unit B 170.3 (C-1), 154.1 (C-7), 130.9 (C-5), 129.6 (C-4), 129.2 (C-9), 122.6 (C.6), 112.3 (C-8) 56.1 (7-OMe), 54.3 (C-2), 35.3 (C-3); unit C 177.8 (C-1), 46.5 (C-3), 42.8 (C-2), 22.9 (2-Me), 23.0 (C-2-Me '); unit D 170.3 (C-1), 71.3 (C-2), 39.7 (C-3), 24.8 (C-4), 22.7 (4-Me), 21.6 (C-5).

 The corresponding diol, Cryptophycin 56 (2.8 mg, 44% yield) was also obtained.

Example 11. Synthesis of Cryptophycin 57
A small amount of PtO ₂ (about 1 mg) was added to the flask containing 0.5 ml of CH ₂ HCl ₂ . The air was removed from the flask and H ₂ was introduced and the mixture was stirred at room temperature for 20 minutes. A solution containing 10.2 mg of Cryptophycin 52 (0.015 mmol) in CH ₂ HCl ₂ (0.3 ml) was added, and the mixture was stirred at room temperature for 20 minutes. The catalyst was removed by filtration through celite / cotton and the solvent was removed in vacuo. Reverse-phase liquid phase gel chromatography (ODS, 10 microns, 250 x 22.5 mm, MeCN / H ₂ O (3: 1), 5 ml min ^-1 ) gave pure Cryptophycin 57 (9.1 mg, 89% yield).

[α] _D +3.4 (c = 4.5, CHCl ₃ ); EIMS m / z 670/672 (M ⁺ , 9/3), 447 (10), 246 (63), 229 (20), 195/197 (78/25), 184 (58), 155/157 (39 / 13), 128 (21), 91 (100), 77 (23); HREIMS m / z 670.3037 (C ₃₆ H ₄₇ N ₂ O ₈ ³⁵ Cl, Δ-1.6 mmu); UV (MeOH) λ _max (ε) 204 (31400), 218 (12000), 284 (1200) nm; ¹ H NMR (CDCl ₃ ) δ unit A: 7.30-7.37 (11/12/13-H, bm), 7.23 (10/14-H, bdd, 7.9, 1.9), 5.03 (5-H, ddd, 9.0 , 5.6, 3.4), 3.66 (8-H, d, 2.1). 2.89 (7-H, dd, 7.7, 2.1), 2.27 (2-H _b , ddd, 14.3, 8.7, 6.2), 2.04 (2-H _a , ddd, 14.3, 8.8, 6.8), 1.64-1.75 (6-H / _4-H2, bm), 1.61 (3-H _{2, bm, W 1/2 ≈25),} 1.11 ( _6-CH3, d, 7.1), unit B: 7.19 (5-H , d, 2.1), 7.04 (9-H, dd, 8.3, 2.1), 6.83 (8-H, d, 8.3), 5.55 (2-NH, d, 8.3), 4.65 (2-H, ddd, 8.3 , 7.3, 5.3), 3.87 (7-OCH ₃ , s), 3.16 (3-Н _b , dd, 14.3, 7.3), 3.08 (3-Н _a , dd, 14.3, 5.3), unit С: 6.91 (3 -NH, dd, 6.4, 6.4), 3.41 (3-H _b , dd, 13.5, 6.4), 3.30 (3-H _a , dd, 13.5, 6.4), 1.21 (2-CH ₃ , s), 1.13 ( 2-CH ₂ ', s), unit D: 4.80 (2-H, dd, 9.8, 4.1), 1.64-1.75 (3-H _b / 4-H, bm), 1.34 (3-H _a , ddd, 15.4, 10.1, 4.1), 0.86 (4-CH ₃ , d, 6.5), 0.84 (5-H ₃ , d, 6.5); ¹³ C NMR (CDCl ₃ ) δ unit A: 172.6 (1), 136.9 (9), 128.7 (11/13), 128.5 (12), 125.6 (10/14), 76.8 (5), 63.4 (7), 59.2 (8), 40.2 (6), 36.2 (2), 32.2 (4), 21.4 (3), 13.6 (6-Me), unit B: 170.4 (1), 154.0 (7), 131.1 (5), 130.0 (4), 128.5 (9), 122.5 (6), 112.2 (8), 56.1 (7-OMe), 54.3 (2), 35.3 (3), unit C: 177.6 (1), 47.0 (3), 43.1 (2), 22.5 (2-Me '), 22.4 (2-Me), unit D: 171.7 (1), 72.0 (2), 39.0 (3), 24.6 (4), 22.8 (4-Me), 21.8 (5).

Example 12. Synthesis of Cryptophycin 58
To a stirred solution of Cryptophycin 57 (5.5 mg, 0.008 mmol) in 3 ml of ethanol-free chloroform at a temperature of about -60 ^° C was added TMSCI (used in the form obtained from Aldrich, about 4.5 mg, about 5.2 μl, about 0.04 mmol). The reaction mixture was stirred for 20 minutes, during which time no starting material remained. Then the volatile components removed under reduced pressure left an amorphous solid. This material was absorbed with acetonitrile and subjected to liquid gel permeation chromatography (ODS, 10 microns, 250 x 22.5 mm, MeCN / H ₂ O (3: 1), 5 ml min ^-1 ) to isolate pure Cryptophycin 58 (5.4 mg yield 93%) as the main product.

-4.9 mmu); UV (MeOH) λ_max(ε) 204 (331900), 218 (11800), 284 (1800) nm; ¹H NMR (CDCl₃) δ unit A: 7.34-7.42 (10/11/12/13/14-Н, bm), 5.01 (5-Н, ddd, 9.6, 8.3. 2.5), 4.65 (8-Н, d, 9.6), 4.00 (7-H, dd, 9.6, 1.9), 2.42 (6.Н, ddq, 8.3, 1.9, 7.0), 2.29 (2-Н_b, ddd, 14.3. 9.4, 4.5), 2.06 (2-H_a, ddd, 14.3, 8.3, 7.5), 1.62-1.82 (3-H₂/4-H₂, bm), 0.99 (6-СН₃, d, 7.0), unit В: 7.20 (5-H, d, 2.1), 7.06 (9-Н, dd, 8.3, 2.1), 6.84 (8-Н, d, 8.3), 5.62 (2-NH, d, 8.3), 4.61 (2-Н, ddd, 8.3, 7.7, 5.4), 3.87 (7-ОСН₃, s), 3.17 (3-Н_b, dd, 14.3, 7.7), 3.11 (3-H_a, dd, 14.3, 5.4), unit C: 6.97 (3-NH, dd, 6.4. 6.2), 3.43 (3-Н_b, dd, 13.4, 6.2), 3.31 (3-Н_a, dd, 13.4, 6.4), 1.23 (2-СН₃, s), 1.16 (2-СН₃', s), unit D: 4.93 (2-Н, dd, 10.0, 4.0), 1.86 (3-H_b, ddd, 14.0, 10.0, 5.5), 1.58 (3-H_a, ddd, 14.0, 8.3, 4.0), 0.97 (4-CH₃, d. 6.8), 0.94 (5-Н₃, d, 6.6); ¹³C NMR (CDCl₃) δ unit A: 172.8 (1), 138.7 (9), 129.0 (12), 128.9 (11/13), 128.0 (10/14), 76.5 (5), 73.8 (7), 62.1 (8), 38.1 (6), 35.9 (2), 31.8 (4), 21.4 (3), 8.7 (6-Me), unit В: 170.6 (1), 153.9 (7), 131.0 (5), 130.2 (4), 128.5 (9), 122.4 (6), 112.2 (8), 56.1 (7-ОМе), 54.4 (2), 35.0 (3), unit С: 177.2 (1), 47.0 (3), 43.2 (2), 22.5 (2-Ме'), 22.4 (2-Me), unit D: 171.8 (1), 72.0 (2), 39.4 (3), 24.9 (4), 22.9 (4-Мe), 21.7 (5).[α] _D +7.2 (c = 2.1, CHCl ₃ ); EIMS m / z 706/708/710 (M ⁺ , 27/23/8), 670/672 (M ⁺ -HCl, 14/13), 583 (54), 581 (53), 485 (23), 483 (21), 447 (34), 294 (21), 282 (39), 246 (57), 195/197 (87/27), 184 (73), 155/157 (45/10), 128 (30 ), 91 (95), 77 (30), 69 (100); HREIMS m / z 706.2844 (C ₃₆ H ₄₈ N ₂ O ₈ ³⁵ Cl ₂ , Δ-5.6 mmu), m / z 670.3070 (M ⁺ -HCl, C ₃₆ H ₄₇ N ₂ O ₈ ³⁵ Cl,

-4.9 mmu); UV (MeOH) λ _max (ε) 204 (331900), 218 (11800), 284 (1800) nm; ¹ H NMR (CDCl ₃ ) δ unit A: 7.34-7.42 (10/11/12/13/14-Н, bm), 5.01 (5-Н, ddd, 9.6, 8.3. 2.5), 4.65 (8-Н , d, 9.6), 4.00 (7-H, dd, 9.6, 1.9), 2.42 (6.Н, ddq, 8.3, 1.9, 7.0), 2.29 (2-Н _b , ddd, 14.3. 9.4, 4.5), 2.06 (2-H _a , ddd, 14.3, 8.3, 7.5), 1.62-1.82 (3-H ₂ /4-H ₂ , bm), 0.99 (6-CH ₃ , d, 7.0), unit B: 7.20 ( 5-H, d, 2.1), 7.06 (9-H, dd, 8.3, 2.1), 6.84 (8-H, d, 8.3), 5.62 (2-NH, d, 8.3), 4.61 (2-H, ddd, 8.3, 7.7, 5.4), 3.87 (7-OCH ₃ , s), 3.17 (3-H _b , dd, 14.3, 7.7), 3.11 (3-H _a , dd, 14.3, 5.4), unit C: 6.97 (3-NH, dd, 6.4. 6.2), 3.43 (3-Н _b , dd, 13.4, 6.2), 3.31 (3-Н _a , dd, 13.4, 6.4), 1.23 (2-СН ₃ , s) , 1.16 (2-CH ₃ ', s), unit D: 4.93 (2-H, dd, 10.0, 4.0), 1.86 (3-H _b , ddd, 14.0, 10.0, 5.5), 1.58 (3-H _a , ddd, 14.0, 8.3, 4.0), 0.97 (4-CH ₃ , d. 6.8), 0.94 (5-H ₃ , d, 6.6); ¹³ C NMR (CDCl ₃ ) δ unit A: 172.8 (1), 138.7 (9), 129.0 (12), 128.9 (11/13), 128.0 (10/14), 76.5 (5), 73.8 (7), 62.1 (8), 38.1 (6), 35.9 (2), 31.8 (4), 21.4 (3), 8.7 (6-Me), unit B: 170.6 (1), 153.9 (7), 131.0 (5), 130.2 (4), 128.5 (9), 122.4 (6), 112.2 (8), 56.1 (7-OMe), 54.4 (2), 35.0 (3), unit C: 177.2 (1), 47.0 (3), 43.2 (2), 22.5 (2-Me '), 22.4 (2-Me), unit D: 171.8 (1), 72.0 (2), 39.4 (3), 24.9 (4), 22.9 (4-Me), 21.7 (5).

Example 13. Synthesis of Cryptophycin 61
Triphenylphosphine sulfide (4 mg, 0.014 mmol) was added to a solution of Cryptophycin 53 (5 mg, 0.007 mmol) in dry benzene, followed by 0.65 μl of trifluoroacetic acid in the form of a solution in dry benzene (100 μl). The solution was allowed to stir at room temperature for 6 hours, neutralized with sodium bicarbonate, filtered and evaporated. The residue was partitioned between water and CH ₂ Cl ₂ . The CH ₂ Cl ₂ soluble substance was purified by reverse phase gel permeation chromatography on a C18 column using a 4: 1 MeOH / H ₂ O mixture to give pure Cryptophycin 61 (1.9 mg, 37% yield).

[α] _D +28.4 (c = 0.7, CHCl ₃ ); EIMS m / z 684/686 (M ⁺ , not observed), 652/654 (M ⁺ -S, 5/4), 426/428 (90/29), 294 (10), 227 (100), 195 / 197 (57/20), 184 (20), 155/157 (34/9), 131 (45), 129 (44), 91 (76), 77 (27); HREIMS m / z 652.2973 (M ⁺ -S, C ₃₆ H ₄₅ N ₂ O ₇ ³⁵ Cl, Δ-5.8 mmu); UV (MeOH) λ _max (ε) 204 (26700), 218 (11600), 284 (820) nm; IR (NaCl) ν _max 3410, 3271, 2958, 1749, 1724, 1670, 1503, 1463, 1258, 1176, 1066, 758 cm ^-1 . ¹ H NMR (CDCl ₃ ) δ unit A 7.29-7.34 (11/12/13-H; m), 7.25 (10/14-H, bd, 6.6), 6.73 (3-H, ddd, 15.2 / 10.6 / 4.5), 5.66 (2-Н; dd, 15.2 / 1.7), 5.22 (5-Н, ddd, 11.2 / 4.2 / 2.0), 3.68 (8-Н, d, 5.1), 3.01 (7-Н, dd. 8.4 / 5.1), 2.52 (4-H _b , dddd, -14.4 / 4.5 / 2.0 / 1.7). 2.41 (4-H _a , ddd, -14.4 / 11.2 / 10.6), 1.68-1.74 (6-H, m), 1.14 (6-Me, d, 6.9); unit B 7.18 (5-H, d, 2.2), 7.04 (9-H, dd, 8.4 / 2.2), 6.84 (8-H, d, 8.4), 5.45 (NH, d, 7.8), 4.75 (2- H, ddd, 7.8 / 7.3 / 5.4), 3.87 (OMe, s), 3.09 (3-H _b , dd, -14.5 / 5.4), 3.05 (3-H _a , dd, -14.5 / 7.3); unit C 7.17 (NH, dd, 8.3, 3.9), 3.39 (3-H _b , dd, -13.5 / 8.3), 3.14 (3-H _a , dd, -13.5 / 3.9), 1.23 (2-Me, s ), 1.16 (2-Me ', s); unit D 4.86 (2-H, dd, 10.2 / 3.4), 1.77 (3-H _b , ddd, -14.0 / 10.2 / 4.9), 1.68-1.74 (4-H, m), 1.42 (3-H _a , ddd, -14.0 / 8.7 / 3.4), 0.92 (4-Me, d, 6.6), 0.88 (5-H ₃ , d, 6.4). ¹³ C NMR (CDCl ₃ ) δ unit A 164.9 (1), 141.7 (3), 138.3 (9), 128.8 (11/13), 128.0 (12), 126.7 (10/14), 124.7 (2), 76.6 (5), 45.8 (7), 43.9 (8), 43.9 (6), 36.6 (4), 16.0 (6-Me), unit B 170.2 (1), 154.1 (7), 130.9 (5), 129.4 ( 4), 128.3 (9), 122.8 (6), 112.4 (8), 56.1 (7-OMe), 54.2 (2), 35.3 (3), unit C 177.9 (1), 46.5 (3), 42.7 (2 ), 22.9 (2-Me), 22.8 (2-Me '), unit D 170.4 (1), 71.3 (2), 39.4 (3), 24.7 (4), 22.7 (4-Me), 21.4 (5) .

Example 14 Synthesis of Cryptophycin 81
Compound S
Compound S is tert-butyl diphenylsilyl ether (TBDVS) derivative F.

Compound T
To 10 ml of a THF solution of p-methoxybenzyltriphenylphosphonium chloride (1 mmol) at -78 ^° C, 400 μl of butyl lithium (1 mmol, 2.5 M in hexane) was added. The mixture was stirred for 30 minutes and then a 2.64 ml aliquot of a 3 ml THF aldehyde S solution (75.0 mg, 0.24 mmol) was added at -78 ^° C. After 30 minutes, cooling was stopped, but stirring was continued for an additional two hours, while whose temperature slowly rose to 25 ^° C. The reaction was quenched with a saturated solution of ammonium chloride and the THF was evaporated. The product was extracted into hexane twice and the combined organic extracts were washed with brine, dried and then concentrated. The residue was transferred to a flash column with silica gel (3% EtOAc / Hexane) to give 63 mg of compound T and 40 mg of a mixture of T and Z isomers.

19.5

16.2 (6-СН₃).Compound T had the following properties:
[α] _D +110.5 ^o (CHCl ₃ , s 0.75); IR ν _max 2956, 2857, 1724, 1608, 1511, 1428, 1250, 1173, 1111, 1037, 821, 703, 505 cm ^-1 : EIMS m / z (relative intensity%) 497 (<1, M ⁺ -OMe ), 471 (31, M ⁺ -Bu '), 367 (56), 294 (31), 199 (75), 135 (100); high-resolution EIMS 497.24770 (calcd for C ₃₂ H ₃₇ O ₃ Si, Δ + 3.5 mmu, M ⁺ -OMe), 471.19859 (calcd for C ₂₉ H ₃₁ O ₄ Si, Δ + 0.6 mmu, M ⁺ -Bu ') . ¹ H NMR δ 7.71 / 7.68 (SiPh ₂ , 2'-H, 6'-H / 2 '' - H, 6 '' - H; d; 6.5), 7.45 / 7.43 (SiPh ₂ , 4'-H / 4``-H; t; 7.4), 7.39 / 7.38 (SiPh <sub>2</sub> , 3'-Н, 5'-Н / 3 "-Н, 5``-Н; dd; 7.4. 6.5), 7.24 (10- H, 14-H; d; 8.7), 6.85 (11-H, 13-H; d; 8.7), 6.79 (3-H; dt; 15.7, 7.5), 6.19 (8-H, d, 16.1), 6.00 (7-H, dd, 16.1. 8.1), 5.66 (2-H, dt, 15.7, 1.3), 3.82 (5-H, m), 3.81 (-OSH ₃ , S), 3.69 (CO ₂ CH ₃ , S), 2.41 (6-H, m), 2.36 (4-H, m), 2.30 (4-H, m), 1.12 (6-CH ₃ , d, 7.0), 1.09 (CMe ₃ , S) . ¹³ C NMR δ 166.7 (1), 158.8 (12) 146.0 (3), 136.0 (SiPh _2, 2 ', 6' / 2 '', 6 ''), 134.1 / 133.7 ( SiPh _2, 1 '/ 1``), 130.4 (9), 130.0 (8), 129.7 / 129.6 (SiPh ₂ , 4 '/ 4''), 129.5 (7), 127.6 / 127.5 (3', 5 '/ 3'', 5 ''), 127.1 (10, 14), 122.8 (2), 113.9 (11, 13), 76.4 (5), 55.2 (OCH ₃ ), 51.3 (СО ₂ СН ₃ ), 42.1 (6), 37.1 (4 ), 27.0

19.5

16.2 (6-CH ₃ ).

 Additionally, compound T was obtained from a mixture of T and Z isomers, 40 mg of a mixture of E and Z isomers was dissolved in 4 ml of a benzene solution containing thiophenol (0.002 M) and ACN (0.006 M). The mixture was heated at the boil under reflux for 5 hours. Treatment and purification on a short silica gel column gave 37.2 mg of compound T.

19.5

16.0 (6-СН₃).Compound U
To 6 ml of an acetone solution of compound T (76 mg, 0.15 mmol) was added 4.4 ml of 1 N LiOH in water. The clear solution was stirred overnight. The acetone was evaporated and the aqueous solution was acidified with 1 N HCl. The product was extracted three times with EtOAc. The organic layer was dried and concentrated. Purification on a silica gel column (20% EtOAc / hexane with 0.5% AcOH) afforded 62.2 mg of the acid of compound U (81% yield);
[α] _D +120.8 ^° (CHCl ₃ , s 3.1); IR ν _max 2960, 2858, 1695, 1650, 1511, 1427, 1250, 1111, 1036, 702 cm ^-1 ; ¹ H NMR δ 7.73 / 7.70 (SiPh ₂ , 2'-H, 6'-H / 2 '' - H, 6 '' - H, d, 7.0), 7.50 (SiPh ₂ , 4'-H / 4 ''-H, m), 7.44 ( SiPh _2, 3'-H, 5'-H / 3''- H, 5''- H, m), 7.29 (10-H, 14-H, d, 8.6 ), 6.96 (3-Н; dt; 15.6, 7.8), 6.89 (11-Н, 13-Н, d, 8.6), 6.22 (8-Н, d, 16.0), 6.03 (7-H, dd, 16.0 7.9), 5.70 (2-H, d, 15.6), 3.88 (5-H, m), 3.83 (OCH ₃ , S), 2.43 (6-H, m), 2.40 (4-H, m), 1.17 (6-CH ₃ , d, 6.9); 1.14 (CMe ₃ , s); ¹³ C NMR, 171.7 (1), 158.8 (12), 148.8 (3), 135.0 (SiPh ₂ , 2 ', 6' / 2 '', 6 ''), 133.9 / 133.7 (SiPh ₂ , 1 '/ 1 ''), 130.3 (9), 130.0 (8), 129.7 (SiPh ₂ , 4 '/ 4``), 129.4 (7), 127.6 (SiPh ₂ , 3', 5 '/ 3'',5'' ), 127.1 (10, 14), 122.5 (2), 113.9 (11, 13), 76.2 (5), 55.2 (OCH ₃ ), 42.3 (6), 37.1 (4), 27.0

19.5

16.0 (6-CH ₃ ).

Compound V
Compound U (59 mg, 0.12 mmol), the trifluoroacetate salt of compound I (57.2 mg, 0.12 mmol) and diisopropylethylamine (DIEA, 62 μl, 0.36 mmol) were dissolved in 1.5 ml of dry DMF. Pentafedipheniphosphinate (FDPP, 55 mg, 0.14 mmol) in 0.6 ml of DMF was added to this solution, and the reaction mixture was stirred for two hours. Ether was added and the organic layer was washed successively with 1 N HCl, saturated sodium bicarbonate, and brine, respectively. Concentration and purification by chromatography (silica gel column, 8% EtOAc / Hexane) afforded 74.2 mg of compound V (72% yield).

19.5

16.6 (6-Me); unit В 170.0 (1), 154.2 (7), 131.0 (5), 128.4 (4/9), 122.5 (6), 112.1 (8), 94.2 (ССl₃), 74.7

56.1 (OMe), 52.9 (2) 36.4 (3).[α] _D +53.8 ^° (CHCl ₃ , s 1.6); IR ν _max 3286, 2959, 1760, 1667, 1640, 1607, 1510, 1253, 1174, 1111, 1067, 1027, 703 cm ^-1 ; EIMS m / z (relative intensity%) 798/799/800/801/802/803/804/805 (31/14/44/17/23/10/6/3. M ⁺ -Bu '), 766 ( 40), 694/695/696/697/698/699/700/701 (70/31/100/38/58/19/14/5), 662 (67), 622 (71), 544 (70) 518 (83); high-resolution EIMS 798.1443 (calcd for C ₄₀ H ₄₀ Cl ₄ NO ₆ Si, Δ-6.4 mmu, M ⁺ -Bu '). ¹ H NMR δ unit A 7.69 / 7.65 (SiPh ₂ , 2'-H, 6'-Н / 2 '' - Н, 6 '' - Н; d; 6.5), 7.41 (SiPh ₂ , 4'-H / 4``-H, m), 7.35 (SiPh <sub>2</sub> , 3'-H, 5'-Н / 3 '' - Н, 5``-Н, m), 7.24 (10-Н, 14-Н; d , 8.7), 6.85 (11-Н, 13-Н, d, 8.7), 6.65 (3-Н, dt, 15.3, 7.5), 6.20 (8-Н, d, 16.1), 6.03 (7-Н, dd , 16.1, 8.0), 5.50 (2-H, d, 15.3), 3.81 (OCH ₃ , S), 3.77 (5-H, m), 2.39 (6-H, m), 2.34 (4-H, m ), 2.29 (4-H ', m), 1.11 (6-Me, d, 6.9), 1.06 (CMe ₃ , S); unit B 7.15 (5-H, d, 1.8), 7.00 (9-H, dd, 8.4, 1.8), 6.83 (8-H, d, 8.4), 5.65 (NH, d, 7.7), 5.01 (2- H, ddd, 7.7, 6.0, 5.5), 4.78 / 4.72 (CH ₂ CCl ₃ , ABq, -11.9), 3.86 (OMe, S), 3.15 (3-H; dd, 6.1, -14.5), 3.08 (3 -H ', dd, 5.8, -14.5). ¹³ C NMR δ unit A 165.1 (1), 158.8 (12), 142.5 (3), 136.0 (SiPh ₂ , 2 ', 6' / 2 '', 6 ''), 134.2 / 133.6 (SiPh ₂ , 1 ' / 1``), 130.4 (9), 129.9 (8), 129.7 / 129.6 (SiPh <sub>2</sub> , 4 '/ 4''), 129.5 (7), 127.6 / 127.5 (SiPh <sub>2</sub> , 3', 5 ' / 3``, 5 ''), 127.1 (10, 14), 124.6 (2), 113.9 (11, 13), 76.4 (5), 55.2 (OMe), 42.1 (6), 37.2 (4), 27.0

19.5

16.6 (6-Me); unit B 170.0 (1), 154.2 (7), 131.0 (5), 128.4 (4/9), 122.5 (6), 112.1 (8), 94.2 (CCl ₃ ), 74.7

56.1 (OMe), 52.9 (2) 36.4 (3).

56.1 (OMe), 53.0 (2), 36.5 (3).Compound W
To a solution of compound V (55.8 mg, 0.065 mmol) in 5.7 ml of acetonitrile was added 0.2 ml of 49% hydrofluoride acid at 0 ^° C. An ice bath was removed after five minutes and the reaction mixture was intensively stirred for 17 hours. The product was extracted into ether, washed successively with saturated sodium bicarbonate solution and brine. Concentration and purification using normal phase chromatography (silica gel column, 25% EtOAc / hexane) gave 31.6 mg of compound W (yield 79%);
[α] _D -3.7 ^o (CHCl ₃ , s 1.3); IR ν _max 3286, 2961, 1756, 1668, 1634, 1607, 1510, 1251, 1175, 1066, 812 cm ^-1 . ¹ H NMR δ unit A 7.27 (10-H, 14-H, d, 8.5), 6.93 (3-H, dt, 15.4, 7.6), 6.83 (11-H, 13-H, d, 8.5), 6.34 (8-H, d, 15.9), 5.88 (7-H, dd, 15.9, 8.2), 5.86 (2-H, d, 15.4), 3.81 (OMe, S), 3.80 (5-H, m), 2.40 (6-H, m), 2.36 (4-H, m), 1.13 (6-Me, d, 6.8); unit B 7.17 (5-H, d, 1.9), 7.05 (9-H, dd, 8.5, 1.9), 6.83 (8-H, d, 8.5), 5.90 (NH, d, 7.7), 5.03 (2- H, ddd, 7.8, 5.9, 5.6), 4.79 / 4.72 (CH ₂ CCl ₃ , ABq, -11.9), 3.86 (OCH ₃ , S), 3.20 (3-H, dd, 6.0, -14.3), 3.09 ( 3-H ', dd, 5.9, -14.3), ¹³ C NMR δ unit A 165.2 (1), 159.1 (12), 142.6 (3), 131.3 (9), 129.8 (7), 128.7 (8), 127.3 (10, 14), 125.0 (2), 114.0 (11, 13), 73.8 (5), 55.3 (OMe), 43.3 (6), 37.2 (4), 16.9 (6-Me) unit B 170.1 (1) , 154.2 (7), 131.0 (5), 128.5 (4/9), 122.5 (6), 112.2 (8), 94.2 (CCl ₃ ), 74.7

56.1 (OMe), 53.0 (2), 36.5 (3).

(2S, 2'R) -2- [3 '(tert-butoxycarbonyl) amino-2'-methylpropanoyl-hydroxy] -4-methylpentanoic acid (AC)
A solution of methyl (S) - (+) - 3-hydroxy-2-methylpropanoate (X) (10 g, 85 mmol) in 300 ml of approximately 9 M ammonia in methanol was heated to 50 ^{° C.} in an evacuated glass ampoule for 168 hours, purged with argon to remove excess ammonia and then evaporated to dryness in vacuo. The residue triturated with ether left behind (S) - (+) - 3-hydroxy-2-methylpropanamide (5.7 g, 66% yield) as a white solid, mp 85.5-87.5 ^o С
[α] _D +28.7 ^° (c 3.5, MeOH); EIMS m / z (rel intensity) 88 (19, M-Me), 85 (35), 73 (69); HREIMS m / z 88.0397 (C ₃ H ₆ NO ₂ , Δ + 0.2 mmu); IR ν _max 3384, 2960, 1671, 1473 cm ^-1 ; ¹ H NMR δ 5.83 (NH; br s), 5.42 (NH; br s), 3.73 (3-H ₂ ; m), 2.55 (2-H; m), 1.19 (2-Me; d, 7.2); ¹³ C NMR δ 180.7 (1), 65.4 (3), 44.0 (2), 14.5 (2-Me). Anal. Found: C, 46.45; H, 8.83. Calcd for C ₄ H ₉ NO ₂ : C, 46.59; H, 8.79.

A suspension of (S) - (+) - 3-hydroxy-2-methylpropanamide (2.1 g, 20 mmol) in anhydrous THF (20 ml) was slowly added to 1 M boron-THF complex (61 mmol, 61 ml), cooled to 0 ^o C. The mixture was refluxed for 6 hours, cooled to 0 ^o C, carefully decomposed with concentrated HCl (10 ml) and concentrated in vacuo. The concentrate was saturated with NaOH (20 g), extracted with CHCl ₃ (4 times 15 ml) and the combined extracts were dried (MgSO ₄ ). After filtering and removing the solvent, vacuum distillation afforded 1.4 g (77% yield) of (R) -3-amino-2-methylpropan-1-ol (Y) as a colorless oil with a boiling point. 110-112 ^o With (40 mm Hg).

[α] _D +8.9 ^o (s 22.6, MeOH); IR ν _max 3358, 1873, 1598, 1466 cm ^-1 ; ¹ H NMR δ 5.18 (NH ₂ , br s), 3.8 (1-H ₂ ; m), 2.95 (3-H; m), 2.68 (3-H; m), 1.81 (2-H; m), 0.82 (2-Me; d, 7.2); ¹³ C NMR δ 66.9 (1), 46.4 (3), 37.1 (2), 14.4 (2-Me).

To a solution of amino alcohol Y (2.0 mg, 22 mmol) in 39 ml of a 10% solution of triethylamine in MeOH was added di-tert-butyl dicarbonate (5.4 mg, 25 mmol) and the mixture was stirred at 25 ^° C for 30 minutes. After removing the solvent, the residue was dissolved in CH ₂ Cl ₂ and the solution was washed twice with 1 M KHSO ₄ (pH 2) and once with saturated NaCl solution and dried (MgSO ₄ ). Removal of the solvent in vacuo afforded 4.3 g (100% yield) of (R) -3- (tert-butoxycarbonyl) amino-2-methylpropan-1-ol as a viscous oil, which was used directly in the next step without further purification (more than 95%) purity, according to NMR analysis);
IR ν _max 3356, 1976, 1686, 1523, 1456 cm ^-1 ; ¹ H NMR δ 4.82 (NH; br s), 3.54 (1-H; dd, -11.4 / 4.2), 3.31 (1-H / 3-H; m), 3.25 (3-H; dd, -14.1 / 6.6), 1.77 (2-H; m), 1.44 (CMe ₃ ; s), 0.87 (2-Me; d, 6.9).

To a solution of alcohol (R) -3- (tert-butoxycarbonyl) amino-2-methylpropan-1-ol (2.2 g, 12 mmol) of sodium periodate (7.5 g, 35 mmol) in carbon tetrachloride (25 ml), acetonitrile (25 r) and water (38 ml) were added ruthenium trichloride hydrate (51 mg, 0.25 mmol) and the mixture was stirred at 25 ^° C for 1 hour. The mixture was diluted with CH ₂ Cl ₂ (100 ml) and then filtered through celite. The filtrate was acidified (pH 9) with a 2 M K ₂ CO ₃ solution and the aqueous layer was washed with ether. The aqueous layer was acidified with a solution of 1 M KHSO ₄ to pH 2 at 0 ^o C and was extracted with CH ₂ Cl ₂ (3 times in 20 ml). The combined extracts were washed with saturated NaCl and dried (MgSO ₄ ). Removal of the solvent in vacuo afforded 2.0 g (85% yield) of (R) -3- (tert-butoxycarbonyl) amino-2-methylpropanoic acid (Z) as a sticky solid. Pure Z (1.75 g, yield 74%) was crystallized from ether, mp. 69.5-70.5 ^o C.

79.5/81.0

42.7/44.0 (3), 39.9/40.2 (2), 28.3/28.3

14.6/14.6 (2-Me), Anal. Found: С, 53.04; H, 8.62. Calcd for C₉H₁₇NO₄: С, 53.18; Н, 8.43.[α] _D -18.4 ^° (s 2, MeOH); EIMS m / z (rel intensity) 147 (60; M ⁺ -Me ₂ C = CH ₂ ), 130 (12), 74 (29), 57 (100); HREIMS m / z 147.0517 (C ₅ H ₉ NO ₄ , Δ + 1.4 mmu); IR ν _max 3322-2400, 2797, 1711, 1654, 1413 cm ^-1 ; ¹ H NMR of major conformer δ 5.00 (NH; br s), 3.32 (3-H; m), 3.24 (3-H '; m), 2.71 (2-H; m), 1.44 (CMe ₃ ; s) 1.20 (2-Me; d); ¹³ C NMR of major / minor (2: 1 ratio) conformers δ 180.7 / 179.5 (1), 156.0 / 157.7

79.5 / 81.0

42.7 / 44.0 (3), 39.9 / 40.2 (2), 28.3 / 28.3

14.6 / 14.6 (2-Me), Anal. Found: C, 53.04; H, 8.62. Calcd for C ₉ H ₁₇ NO ₄ : C, 53.18; H, 8.43.

To a solution of 2.66 g of L-leucinic acid (20 mmol) and 1.74 g of sodium bicarbonate (20 mmol) in 30 ml of water at 0 ^{° C.} were added 30 ml of a solution of 6.44 g of tetrabutylammonium chloride (20 mmol) and 1.74 ml of allyl bromide (20 mmol) in CH ₂ Cl ₂ . After vigorously stirring the mixture for 24 hours, CH ₂ Cl _{2 was} evaporated. About 50 ml of water was added and the aqueous layer was extracted four times with Et ₂ O. The ethereal solution was dried over anhydrous sodium sulfate and then evaporated. The residue was passed through a short Si column to give 3.21 g of allyl (2S) -2-hydroxy-4-methylpentanoate (AA) (yield 93%) as a colorless oil,
[α] _D -8.4 ^° (c 1.1, CHCl ₃ ); IR ν _max 3464, 2957, 1732, 1203, 1140, 1087 cm ^-1 ; ¹ H NMR δ 5.92 (allyl 2-H; m), 5.34 (allyl 3-H ₂ ; dd, 17.4 / 1.1), 5.28 (allyl 3-H _E ; dd, 10.5 / 1.1), 4.67 (allyl 1-H ₂ ; d, 5.7), 4.23 (2-H; br s), 2.64 (OH; br s), 1.89 (4-H; m), 1.57 (3-H ₂ ; m), 0.96 (5-H ₃ ; d, 6.5), 0.95 (4-Me; d, 6.7); ¹³ C NMR δ 175.3 (1), 131.4 (allyl C-2), 118.6 (3), 68.9 (2), 65.7 (allyl C-1), 43.2 (3), 24.1 (4), 23.0 (5), 21.3 (4-Me).

5.17 (NH; br s), 3.42 (3-H; m), 3.22 (3-H'; m), 2.78 (2-Н, m), 1.43 (СМе₃; br s), 1.21 (2-Me; d. 7.1); unit D δ 5.90 (allyl 2-H; m), 5.33 (allyl 3-Н_Z; d, 16.3), 5.27 (allyl 3-Н_E; d, 10.3), 5.09 (2-H; dd, 9.7/3.7), 4.63 (allyl 1-H₂; m), 1.80 (3-H₂; m), 1.64 (4-H; m), 0.96 (5-H₃; d, 6.5), 0.94 (4-Me; d, 7.3), ¹³С NMR unit C δ 174.7 (1), 156.0

79.2

43.1 (3), 40.3 (2), 28.3

14.5 (2-Me); unit D δ 170.4 (1), 131.4 (allyl C-2), 119.0 (allyl C-3), 70.9 (2), 65.9 (allyl C-1), 39.6 (3), 24.7 (4), 23.0 (5), 21.5 (4-Me).To a solution of 1.74 g Z (8.55 mmol), 1.34 g AA (8.0 mmol) and 64 mg DMAP in 12 ml of dry CH ₂ Cl ₂ at 0 ^° C were added dropwise 8 ml of a solution of DDC (2.47 g, 12 mmol) in CH ₂ Cl ₂ . The clear solution was stirred at 0 ^{° C.} for 30 minutes and then at 23 ^{° C.} for 3 hours. The white precipitate was filtered off, the solvent was evaporated and the residue was redissolved in Et ₂ O. The ether solution was washed successively with cold 0.5 N NaCl solution, sodium bicarbonate solution and brine. The dried ether layer (Na ₂ SO ₄ ) was evaporated and the product was purified using flash chromatography (silica gel) to give 2.62 g (yield 92%) of pure allyl (2S, 2'R) -2- [3 '(tert-butoxycarbonyl) amino-2'-methylpropanoyl-hydroxy] -4-methylpentanoate (AB) as a colorless oil,
[α] _D -51.3 ^° (c 3.41, CHCl ₃ ); EIMS m / z (rel intensity) 301 (5.2), 284 (4.0), 258 (1.5), 228 (43.5), 170 (41.8), 130 (74.5), 112 (100); HREIMS m / z, 301.1532 (C ₁₄ H ₂₃ NO ₆ , Δ-0.7 mmu, M-Me ₂ C = CH ₂ ), 284.1496 (C ₁₄ H ₂₂ NO ₅ , Δ + 0.2 mmu); IR ν _max 3395, 2962, 1747, 1715, 1515, 1251, 1175, 1083 cm ^-1 . ¹ H NMR unit C

5.17 (NH; br s), 3.42 (3-H; m), 3.22 (3-H '; m), 2.78 (2-H, m), 1.43 (CMe ₃ ; br s), 1.21 (2-Me ; d. 7.1); unit D δ 5.90 (allyl 2-H; m), 5.33 (allyl 3-H _Z ; d, 16.3), 5.27 (allyl 3-H _E ; d, 10.3), 5.09 (2-H; dd, 9.7 / 3.7 ), 4.63 (allyl 1-H ₂ ; m), 1.80 (3-H ₂ ; m), 1.64 (4-H; m), 0.96 (5-H ₃ ; d, 6.5), 0.94 (4-Me; d, 7.3), ¹³ C NMR unit C δ 174.7 (1), 156.0

79.2

43.1 (3), 40.3 (2), 28.3

14.5 (2-Me); unit D δ 170.4 (1), 131.4 (allyl C-2), 119.0 (allyl C-3), 70.9 (2), 65.9 (allyl C-1), 39.6 (3), 24.7 (4), 23.0 (5 ), 21.5 (4-Me).

1.21 (2-Me; d, 7.1); unit D δ 5.12 (2-H; dd, 9.6/3.5), 1.90-1.68 (3Н/4-Н; m), 0.97 (5-Н; d, 6.1), 0.94 (4-Me; d, 6.0). ¹³C NMR unit С δ 174.6 or 174.8 (1), 156.1

79.5

43.0 (3), 40.4 (2), 28.3

14.5 (2-Me): unit D δ 174.6 or 174.8 (1), 70.5 (2), 39.5 (3), 24.7 (4), 23.0 (5), 21.4 (4-Me).688 μl (8 mmol) of dry morpholine was slowly added to 10 ml of a solution of 282 mg (0.8 mmol) AB and 91 mg (0.08 mmol) of tetrakis (triphenylphosphine) palladium in dry THF. After stirring for 40 minutes, the solvent was evaporated and 100 ml of CH ₂ Cl _{2 was} added. The solution was washed successively with 2 N HCl (40 ml) and water. The organic layer was filtered and the filtrate was extracted twice with saturated sodium bicarbonate solution. After backwashing with CH ₂ Cl ₂ , the aqueous layer was first acidified to pH 3 with a cold KHSO ₄ solution at 0 ^{° C.} and then extracted three times with ether. The dried ether extract was evaporated to give 250 mg of (2S, 2'R) -2- [3 '(tert-butoxycarbonyl) amino-2'-methylpropanoyloxy] -4-methylpentanoic acid (AC) (yield 100%) as a wax-like solid
[α] _D -47.9 ^° (c 4.7, CHCl ₃ ); EIMS m / z (rel intensity) 261 (12), 244 (18), 217 (28), 198 (17), 188 (100), 160 (61); HREIMS m / z 261.1221 (С ₁₁ Н ₁₉ NO ₆ , Δ-0.8 mmu, М-Ме ₂ C = СН ₂ ), 244.1221 (C ₁₁ H ₁₈ NO ₅ , Δ-3.6 mmu); IR ν _max 3376, 2960, 1738, 1518, 1174, 786 cm ^-1 , ¹ H NMR (CDCl ₃ + D ₂ O) unit С δ 3.49 (H-3; dd, -13.8 / 3.5), 3.12 (3- H; dd, -13.8 / 8.7), 2.68 (2-H; m), 1.43

1.21 (2-Me; d, 7.1); unit D δ 5.12 (2-H; dd, 9.6 / 3.5), 1.90-1.68 (3Н / 4-Н; m), 0.97 (5-Н; d, 6.1), 0.94 (4-Me; d, 6.0) . ¹³ C NMR unit C δ 174.6 or 174.8 (1), 156.1

79.5

43.0 (3), 40.4 (2), 28.3

14.5 (2-Me): unit D δ 174.6 or 174.8 (1), 70.5 (2), 39.5 (3), 24.7 (4), 23.0 (5), 21.4 (4-Me).

56.1 (ОМе), 53.2 (2), 36.6 (3); unit С 175.2 (1), 156.0

79.3

43.1 (3), 40.4 (2), 28.3

14.4 (2-Me); unit D 170,1 (1), 71.4 (2), 39.5 (3), 24.7 (4), 22.9 (5), 21.4 (4-Ме).Compound AD
Alcohol W (34.8 mg, 0.056 mmol), compound AC (26.8 mg, 0.085 mmol) and DMAP (1.74 mg) were dissolved in 283 μl of dichloromethane. To this mixture was added 666 μl of a solution of DDC (17.5 mg, 0.085 mmol) in dichloromethane. The reaction mixture was stirred overnight. The solvent was evaporated with a stream of nitrogen, ether was added and a white precipitate was filtered off. The filtrate was washed successively with 0.5 N HCl, saturated sodium bicarbonate and brine. Concentration and chromatography (silica gel column, 25% EtOAc / Hexane) afforded 46 mg of compound AD (90% yield);
[α] _D -11.8 ^° (CHCl ₃ , s 2.0); IR ν _max 3369, 2961, 1737, 1511, 1252, 1174, 1066, 813, 756 cm ^-1 . ¹ H NMR (500 MHz) δ unit A 7.25 (10-H / 14-H, dt, 8.5. 2.1), 6.84 (11-H / 13-H, dt, 8.5, 2.1), 6.76 (3-H, ddd, 15.5, 6.5, 6.4), 6.34 (8-H, d, 15.6), 5.88 (2-H, bd, 15.5), 5.86 (7-H, dd, 15.6, 8.7), 5.04 (5, m) 3.80 (OMe, s), 2.56 (6-H, m), 2.52 (4-H, m), 1.10 (6-Me, d, 6.8); unit B 7.19 (5-H, d, 2.1), 7.05 (9-H, dd, 8.5, 2.1), 6.83 (8-H, d, 8.5), 6.55 (NH, bd, 7.3), 5.04 (2- H, m), 4.78 / 4.70 (CH ₂ CCl ₃ , ABq, -11.8), 3.85 (OCH ₃ , S), 3.19 (3-Н, dd, 6.3, -13.8), 3.08 (3-Н ', dd 6.8. -13.8); unit C 5.14 (NH, bt, 6.3), 3.32 (3-H ', m), 3.20 (3-H', m), 2.73 (2-H, m), 1.42 (CMe ₃ , s), 1.18 (2 -Me, d, 7.0); unit D 4.93 (2-H, dd, 10.0, 3.7), 1.67 (3-H / 4-H, m), 1.55 (3-H ', m), 0.86 (5-H, d, 6.5) 0.83 (4-Me-H, d, 6.5). ¹³ C NMR δ unit A 165.4 (1), 159.1 (12), 139.3 (3), 131.1 (9), 129.7 (7), 128.5 (8), 127.3 (10, 14), 125.4 (2), 114.0 ( 11, 13), 76.5 (5), 55.3 (OMe), 41.1 (6), 33.4 (4), 16.7 (6-Me); unit B 170.5 (1), 154.1 (7), 131.2 (5), 128.9 (4), 128.5 (9), 122.4 (6), 112.1 (8), 94.3 (CCl ₃ ), 74.6

56.1 (OMe), 53.2 (2), 36.6 (3); unit C 175.2 (1), 156.0

79.3

43.1 (3), 40.4 (2), 28.3

14.4 (2-Me); unit D 170.1 (1), 71.4 (2), 39.5 (3), 24.7 (4), 22.9 (5), 21.4 (4-Me).

Cryptofinin 81
Compound AD (46 mg, 0.05 mmol) was mixed with Zn-activated dust (178 mg, excess) in 1.3 ml of HOAc. The mixture was sonicated for 45 minutes and then stirred for an additional 90 minutes. About 30 ml of dichloromethane was added. The solid was filtered off and the filtrate was evaporated in vacuo. The residue was dissolved in 1.1 ml of TFA and the solution was stirred for one hour. TFA was evaporated in vacuo and water was added. Freeze drying gave the free amino acid. The amino acid was dissolved in 4.6 ml of DMF. To this solution was added 26 μl of DIEA and FDPP (30 mg, 0.075 mmol, in 2.2 ml of DMF), respectively. After stirring for 6 hours, the solvent was evaporated and EtOAc was added. The solution was washed with 0.5 N HCl and brine, respectively. Evaporation of the solvent followed by chromatographic purification (silica gel, ether) gave 20.5 mg of Cryptophycin 81 (61% yield).

[α] _D +34.9 ^° (CHCl ₃ , s 0.45); IR ν _max 3409, 3270, 2958, 1746, 1725, 1672, 1511, 1251, 1175, 1066, 1025, 972, 816 cm ^-1 . ¹ H NMR (500 MHz) δ unit A 7.26 (10-Н / 14-Н; dt, 8.6, 2.5), 6.84 (11-Н / 13-Н, dt, 8.6, 2.5), 6.68 (3-H, ddd, 15.4, 9.9, 5.6), 6.35 (8-H, d, 15.9), 5.86 (7-H, dd, 15.9, 8.8), 5.77 (2-H, dd, 15.4, 0.9), 4.99 (5- H, ddd, 11.2, 6.0, 1.7), 3.80 (OCH ₃ , S), 2.53 (4-Н / 6-Н, m), 2.37 (4-Н ', ddd, 11.2, 9.9, -14.6), 1.12 (6-Me, d, 6.9); unit B 7.22 (5-H, d, 2.2), 7.08 (9-H, dd, 8.4, 2.4), 6.84 (8-H, d, 8.4), 5.64 (NH, d, 8.6), 4.81 (2- H, m), 3.86 (OMe, S), 3.13 (3-H; dd, 5.6, -14.5), 3.05 (3-H ', dd, 7.1, -14.5); unit C, 6.93 (NH, bdd, 5.8, 5.6), 3.50 (3-H, ddd, 5.2, 3.9, -13.5), 3.28 (3-H ', ddd, 6.9, 6.7, -13.5), 2.71 (2 -H, m) 1.22 (2-Me; d, 7.3); unit D 4.84 (2-H, dd, 10.1, 3.4), 1.67 (3-H / 4-H; m), 1.38 (3-H ', m), 0.78 (5-H, d, 6.5), 0.75 (4-Me-H, d, 6.5). ¹³ C NMR (125 MHz) δ unit A 165.4 (1), 159.2 (12), 141.4 (3), 131.1 (9), 129.6 (7), 128.4 (8), 127.3 (10, 14), 125.2 (2 ), 114.1 (11, 13), 77.5 (5), 55.3 (OMe), 42.2 (6), 36.4 (4), 17.4 (6-Me): unit B 171.0 (1), 154.0 (7), 131.2 ( 5), 129.9 (4), 128.4 (9), 122.5 (6), 112.1 (8), 56.2 (OMe), 53.5 (2), 35.1 (3); unit C 175.6 (1), 41.2 (3), 38.3 (2), 14.0 (2-Me); unit D 170.9 (1), 71.6 (2), 39.5 (3), 24.5 (4), 22.7 (5), 21.3 (4-Me).

Example 15. Synthesis of Cryptophycin 82
Compound AE
Compound AE is a derivative of tert-butyl diphenylsilyl ether (TBDMS) E.

AF compound
Hydrolysis of AE (150 mg) was carried out in 87% yield using the procedure described above for the hydrolysis of T and U.

¹ H NMR δ 7.69 / 7.64 (SiPh ₂ , 2'-H, 6'-H / 2 '' - H, 6 '' - H, d, 6.3), 7.41 (SiPh ₂ , 4'-H / 4 ''-H, m), 7.39 (SiPh ₂ , 3'-H, 4-Me-H / 3''- Н, 5``-Н, m), 6.86 (3-H; dt; 15.5, 7.5) 5.62 (2-H, d, 15.5); 5.30 (7-H / 8-H, m).

AG compound
Obtaining AG (96 mg) from AF (76 mg, 0.18 mmol) was performed in 70% yield using the procedure described above to obtain V from U.

19.5

18.1 (9), 16.4/16.3 (6-Ме); unit В 170.0 (1), 154.2 (7), 131.0 (5), 128.4 (4/9), 122.5 (6), 112.1 (8), 94.2 (ССl₃), 74.7

56.1 (OMe), 52.9 (2) 36.4 (3). ¹ H NMR δ unit A 7.67 (SiPh ₂ , 2'-H, 6'-H / 2 '' - H, 6 '' - H; m), 7.46-7.31 (SiPh ₂ , 3'-H, 4 ' -H, 4-Me-H / 3 '' - H, 4 '' - H, 5 '' - H, m), 6.62 (3-H, dt, 15.4, 7.6), 5.51 / 5.48 (2-H , d, 15.4), 5.33 (7/8, m), 3.70 (5-Н, m), 2.22 (4-Н / 6-Н, m), 1.61 (9, bd, 7.4), 1.06 (CMe ₂ , S), 0.98 (6-Me, d, 6.8); unit B 7.16 (5-H, d, 1.7), 7.00 (9-H, dd, 8.5, 1.7), 6.83 / 6.82 (8-H,
d, 8.5), 5.68 / 5.66 (NH, d, 7.3), 5.03 (2-H, m), 4.78 / 4.73 (CH ₂ CCl ₃ , ABq, -11.9), 3.87 (OMe, S), 3.16 (3 -H; m), 3.09 (3-H ', m). ¹³ C NMR δ unit A 165.1 (1), 143.0 / 142.9 (3), 136.0 (SiPh ₂ , 2 ', 6' / 2``, 6 ''), 134.3 / 133.7 (SiPh <sub>2</sub> , 1 '/ 1''), 132.5 (7), 129.6 (8), 129.6 (SiPh <sub>2</sub> , 4' / 4``), 127.5 (SiPh ₂ , 3 ', 4-Me / 3'', 5``), 125.7 / 125.4 (2), 76.3 (5), 41.6 (6), 36.9 / 36.8 (4), 27.0

19.5

18.1 (9), 16.4 / 16.3 (6-Me); unit B 170.0 (1), 154.2 (7), 131.0 (5), 128.4 (4/9), 122.5 (6), 112.1 (8), 94.2 (CCl ₃ ), 74.7

56.1 (OMe), 52.9 (2) 36.4 (3).

Compound AN
The preparation of AN (53.4 mg) from AG (84 mg, 0.11 mmol) was performed in 92% yield using the procedure described above to obtain W from V.

56.1 (OMe), 53.1 (2), 36.5 (3). ¹ H NMR δ unit A 6.83 (3-H, dt, 15.4. 7.5), 5.80 (2-H, d, 15.4), 5.51 (8-H, m), 5.33 (7-H, dd, 15.2 , 8.3), 3.50 (5-H, m), 2.42 (4-H, m), 2.30 (4-H ', m), 2.28 (6, m), 1.68 (9, d, 7.3), 0.99 ( 6-Me, d, 6.7); unit B 7.20 (5-H, d, 1.8), 7.03 (9-H, dd, 8.4, 1.8), 6.80 (8-H, d, 8.4), 6.10 (NH, bd, 7.0), 5.07 (2- H, m), 4.80 / 4.70 (CH ₂ CCl ₃ , ABq, -11.5), 3.88 (OCH ₃ , S), 3.20 (3-Н, dd, 5.5, -14.3), 3.10 (3-Н ', dd , 7.2, -14.3). ¹³ C NMR δ unit A 165.4 (1), 142.8 (3), 132.2 (7), 127.7 (8), 125.7 (2), 73.6 (5), 42.8 (6), 36.9 (4), 18.1 (9) 16.8 (6-Me); unit B 170.2 (1), 154.2 (7), 131.0 (5), 128.5 (4/9), 122.5 (6), 112.2 (8), 94.2 (CCl ₃ ), 74.7

56.1 (OMe), 53.1 (2), 36.5 (3).

Cryptophycin 82
Compound A1 (36.8 mg, 86% yield) was prepared from 27.4 mg (0.052 mmol) of AH using the procedure described above for AD.

Compound A1 (68 mg, 0.083 mmol) was cyclized to Cryptophycin 82 (28.5 mg) using the procedure described above for cyclization of AD to Cryptophycin 81;
[α] _D +19.9 ^° (CHCl ₃ , s 2.0). ¹ N NMR (500 MHz) δ unit A 6.65 (3-H, ddd, 15.4, 9.5, 5.6), 5.76 (2-H, d, 15.4), 5.48 (8-H, dq, 15.3, 6.5), 5.27 (7-Н, ddd, 15.3, 8.4, 1.5), 4.89 (5-Н, ddd, 10.7, 5.6, 1.5), 2.38 (4-Н, m), 2.33 (4-Н '/ 6-Н, m ), 1.66 (9-H, dd, 6.4, 1.5), 1.00 (6-Me, d, 6.9); unit B 7.22 (5-H, d, 2.0), 7.08 (9-H, dd, 8.4, 2.0), 6.83 (8-H, d, 8.4), 5.74 (NH, m), 4.81 (2-H, ddd, 8.5, 7.3, 5.6), 3.87 (OMe, S), 3.13 (3-H; dd, 5.6, -14.4), 3.04 (3-H ', dd, 7.3, -14.4); unit C, 6.93 (NH, bdd, 6.7, 4.7), 3.52 (3-H, ddd, 4.7, 3.9, -13.5), 3.27 (3-H ', ddd, 6.8, 6.7. -13.5), 2.72 (2 -H, dqd, 7.1, 6.8, 3.9), 1.20 (2-Me; d, 7.1); unit D 4.88 (2-H, dd, 9.6, 3.4), 1.75 (3-H / 4-H; m), 1.47 (3-H ', m), 0.94 (5-H, d, 6.2), 0.91 (4-Me-H, d, 6.4). ¹³ C NMR (125 MHz) δ unit A 165.5 (1), 141.7 (3), 131.4 (7), 127.1 (8), 125.2 (2), 77.7 (5), 41.5 (6), 36.2 (4), 17.9 (9), 17.2 (6-Me); unit B 171.0 (1), 153.9 (7), 131.1 (5), 129.9 (4), 128.4 (9), 122.4 (6), 112.2 (8), 56.1 (OMe), 53.5 (2), 35.1 (3 ); unit C 175.5 (1), 41.3 (3), 38.2 (2), 14.0 (2-Me); unit D 170.9 (1), 71.5 (2), 39.6 (3), 24.6 (4), 23.0 (5), 21.5 (4-Me).

Example 16. Synthesis of Cryptophycin 90 and Cryptophycin 91
General procedure for the epoxidation of styrene-type cryptophycins.

Three equivalents of m-chlorobenzoic acid in dichloromethane (about 10 mg / ml) were added to a solution of cryptophycin (about 10 mg / ml) in dichloromethane. The solution was stirred at room temperature until all starting material had reacted. The solution was passed through a short silica gel column using CH ₂ Cl ₂ and then 1: 4 CH ₂ Cl ₂ : Et ₂ O to give a mixture of two epoxides. Epoxides were separated using liquid gel permeation chromatography (C-18, 7: 3 MeCN: H ₂ O).

 Using this procedure, Cryptophycin 82 (5 mg) was converted to 2.2 mg of Cryptophycin 90 and 1.2 mg of Cryptophycin 91.

Spectral data for Cryptophycin 90
¹ H-NMR (500 MHz) δ-link A
6.67 (3-H, ddd, 15.4, 9.5, 5.8), 5.79 (2-H, d, 15.4), 5.09 (5-H, ddd, 10.4, 4.5, 2.6), 2.84 (8-H, qd, 5.2 , 2.2), 2.57 (7-H, dd, 7.6, 2.2), 2.46 (4-H, m), 1.82 (6-H, m), 1.32 (9-H, d, 5.2), 1.03 (6- Me, d, 6.9); unit B 7.22 (5-H, d, 2.2), 7.08 (9-H, dd, 8.4, 2.2), 6.84 (8-H, d, 8.4), 5.74 (NH, d, 8.6), 4.81 (2- H, ddd, 8.3, 7.4, 5.7), 3.87 (OMe, S), 3.14 (3-H; dd, 5.4, -14.5), 3.03 (3-H ', dd, 7.3, -14.5); unit C, 6.95 (NH, bdd, 6.7, 4.8), 3.52 (3-H, ddd, 4.8, 3.7, -13.4), 3.29 (3-H ', ddd, 6.7, 6.5, -13.4), 2.74 (2 -H, dqd, 7.3, 6.5, 3.7), 1.24 (2-Me; d, 7.3); unit D 4.90 (2-H, dd, 9.9, 3.7), 1.75 (3-H, m), 1.60 (4-H; m), 1.49 (3-H ', ddd, 8.7, 3.7, -13.7), 0.95 (5-H, d, 6.5), 0.91 (4-Me-H, d, 6.7). ¹³ C NMR (125 MHz) δ unit A 165.4 (1), 141.1 (3), 125.3 (2), 76.3 (5), 60.0 (7), 56.8 (8), 40.2 (6), 36.5 (4), 17.5 (9), 13.3 (6-Me); unit B 171.0 (1), 154.0 (7), 131.1 (5), 129.9 (4), 128.4 (9), 122.5 (6), 112.3 (8), 56.1 (OMe), 53.6 (2), 35.1 (3 ); unit C 175.5 (1), 41.2 (3), 38.3 (2), 14.1 (2-Me); unit D 170.7 (1), 71.4 (2), 39.6 (3), 24.7 (4), 23.0 (5), 21.5 (4-Me)
Spectral data for Cryptophycin 91
¹ H-NMR (500 MHz) δ-link A
6.68 (3-H, ddd, 15.4, 9.8, 5.4), 5.78 (2-H, d, 15.4), 5.09 (5-H, ddd, 11.1, 3.7, 2.0), 2.75 (8-H, m), 2.62 (7-H, dd, 9.5, 2.1), 2.48 (4-H, m), 1.78 (6-H, m), 1.32 (9-H, d, 5.2), 0.99 (6-Me, d, 7.1); unit B 7.23 (5-H, d, 1.9), 7.09 (9.H, dd, 8.4, 1.9), 6.85 (8-H, d, 8.4), 5.68 (NH, d, 8.4), 4.82 (2- H. m), 3.88 (OMe, S), 3.15 (3-H; dd, 5.4, -14.4), 3.04 (3-H ', dd, 7.2, -14.4); unit C, 6.95 (NH, bdd, 6.0, 4.6), 3.53 (3-H, ddd, 5.0, 4.0, -13.4), 3.29 (3-H ', ddd, 6.9, 6.7, -13.4), 2.75 (2 -H, m), 1.23 (2-Me; d, 6.7); unit D 4.92 (2-H, dd, 9.8, 3.4), 1.77 (3-H, m), 1.58 (4-H; m), 0.96 (5-H, d, 7.3), 0.92 (4-Me- H, d, 6.7).

Example 17. Synthesis of Cryptophycin 97
Zinc azide (40 mg) and concentrated sulfuric acid (4 μl) were added to a solution of the cyclic depsipeptide, Cryptophycin 53 (9 mg, 0.013 mmol) dissolved in dimethyl sulfoxide (1 ml). Then the mixture was allowed to mix at 75-85 ^o C for 2 days. After this time, the reaction mixture was cooled to room temperature, diluted with EtzO (15 ml) and the organic layer was washed with brine (2 times 20 ml) and water (20 ml). Then the ether extract was dried (MgSO ₄ ) and the solvent was removed in vacuo to return an amorphous colorless solid, which was predominantly an azido alcohol, Cryptophycin 86. Product purification was achieved by reverse phase chromatography (ODS, 10 μl, 250 • 1 mm, 25 % H ₂ O / MeCN, 3 ml min ^-1 ) with the return of pure azidoalcohol, Cryptophycin 86, as an amorphous colorless solid (7.4 mg, 77% yield).

Spectral data for Cryptophycin 86
[α] _D -22.0 (c = 3.0, CHCl ₃ ); MS (EI) m / z 482/484 (highest observed ion, M ⁺ -229, 8/3), 625/627 (14/5), 259 (7), 195/197 (100/34), 184 ( 14), 155/157 (82/70), 91 (23), 77 (22); HRMS, obsd m / z 482.1791, C ₂₃ H ₃₁ N ₂ O ₇ ³⁵ Cl (Δ 2.9 mmu); MS (FAB) m / z (magic bullet matrix) 712/714 (M ⁺⁺ H, 79/36), 686/688 (31/12), 232 (74), 184 (100). ¹ H NMR (CDCl ₃ ) δ unit A: 7.37-7.42 (10/11/12/13/14-Н, bm, W _1/2 ≈20), 6.77 (3-H, ddd, 15.2, 10.6, 4.4 ), 5.77 (2-H, dd, 15.2, 1.3), 5.45 (5-H, ddd, 11.0, 4.2, 2.0), 4.55 (8-H, d, 5.7), 3.75 (7-H, dd, 7.3 , 5.7), 2.55 (4-H _b, dddd, 14.5, 4.4, 2.0, 1.3), 2.43 _{(4-H a, ddd, 14.5,} 11.0, 10.6), 2.34 ( 7-OH, s), 1.80 (6 -H, ddq, 7.3, 4.2, 7.0), 0.99 (6-Me, d. 7.0), unit B: 7.20 (5-H, d, 2.2), 7.06 (9-H, dd, 8.4, 2.2), 6.84 (8-H, d, 8.4), 5.76 (NH, d, 7.7), 4.74 (2-H, ddd, 7.7, 7.5, 5.5), 3.87 (OCH ₃ , s), 3.10 (3-H _b , dd, 14.5, 5.5), 3.06 (3-H _a , dd, 14.5, 7.5), unit C: 7.22 (NH, dd, 8.4, 3.7), 3.40 (3-H _b , dd, 13.6, 8.4), 3.14 (3-H _a , dd, 13.6, 3.7), 1.23 (2-CH ₃ , s), 1.16 (2-CH ₃ ', s), unit D: 4.85 (2-H, dd, 9.5, 5.1), 1.71 _{(3-H b, ddd, 13.6,} 9.5, 5.9), 1.59 ( _{4-H, bm, W 1/2 ≈25),} 1.50 ( _{3-H a, ddd, 13.6,} 7.9, 5.1), 0.89 ( 4-CH ₃ , d, 6.6), 0.85 (5-H ₃ , d, 6.6); ¹³ C NMR (CDCl ₃ ) δ unit A: 165.3 (1), 143.0 (3), 135.1 (9), 129.1 (12), 128.9 (11/13), 128.6 (10/14), 124.3 (2), 75.1 (7), 74.6 (5), 67.8 (8), 39.3 (6), 34.7 (4), 11.9 (6-Me), unit B: 170.5 (1), 154.1 (7), 130.9 (5), 129.7 (4), 128.3 (9), 122.5 (6), 112.4 (8), 56.1 (7-OMe), 54.3 (2), 35.3 (3), unit C: 177.8 (1), 46.5 (3), 42.8 (2), 22.9 (2-Me), 22.7 (2-Me '), unit D: 170.2 (1), 71.4 (2), 39.4 (3), 24.7 (4), 22.6 (4-Me), 21.8 (5).

Cryptophycin 97
To a solution of cyclic depsipeptide, Cryptophycin 86 (5.5 mg, 0.008 mmol) dissolved in a mixture of Et ₂ O / CH ₂ Cl ₂ (3: 1 ml, 0.5 ml), an ether solution (0.5 ml) of triphenylphosphine (3 mg, 0.011 mmol). Then the mixture was allowed to mix at room temperature for 3 days. After this time, the solvent was removed in vacuo and the residue was dissolved in CH ₂ Cl ₂ and subjected to purification using liquid gel permeation chromatography (CN column, 10 μm, 250 • 10 mm, 80% EtOAc / CH ₂ Cl ₂ , 3 ml min ^{- 1} ) with the return of pure Cryptophycin 97, in the form of an amorphous colorless solid (4.2 mg, 82% yield).

UV (MeOH) λ _max (ε) 202 (24400), 218 (9400), 284 (2200) nm; MS (EI) m / z 667/669 (M ⁺ , 11/3), 639/641 (41/16), 442 (21), 226 (17), 195 (43), 196 (32), 197 ( 100), 198 (71), 199 (11), 182/184 (25/16), 155/157 (63/22), 146 (30), 91 (40), 77 (29), HRMS, obsd m / z 667.3061, C ₃₆ H ₄₆ N ₃ O ₇ ³⁵ Cl (Δ-3.6 mmu). ¹ H NMR (CDCl ₃ ) δ unit A 7.35 (11-Н / 13-Н; m, W _1/2 ≈15 Hz), 7.28 (12-H, m), 7.16 (10-Н / 14-Н; m, W _1/2 ≈15 Hz), 6.74 (3-H; ddd, 15.2 / 9.0 / 6.1), 5.69 (2-H; d, 15.2), 5.21 (5-H; ddd, 9.2 / 4.3 / 4.2 ), 2.79 (8-H, bs), 2.51 (4-H ₂ , m), 2.11 (7-H, bd, 6.5), 1.48 (6-H, m), 1.13 (6-Me, d, 6.9 ); unit B: 7.18 (5-H, d, 2.1), 7.04 (9-H, d, 8.4, 2.1), 6.83 (8-H, d, 8.4), 5.53 (NH, m), 4.73 (2-H , ddd, 7.6, 5.6, 5.4), 3.87 (OMe, s), 3.09 (3-H _b , dd, 14.7, 5.4), 3.04 (3-H _a , dd, 14.7, 7.6), unit C: 7.20 ( NH, m), 3.40 _{(3-H b, dd, 13.5,} 8.6), 3.11 ( _{3-H a, dd, 13.5,} 3.3), 1.22 ( 2-Me, s), 1.15 (2-Me, s) , unit D: 4.84 (2-H, dd, 10.1, 3.7). 1.71 (3-H _b , m), 1.67 (4.Н, bm, W _1/2 ≈25), 1.35 (3-H _a , m), 0.86 (4-Me, d, 6.7), 0.84 (5 -H ₃ , d, 6.5); ¹³ C NMR (CDCl ₃ ) δ unit A: 165.0 (1), 142.1 (3), 139.2 (9), 128.8 (11/13), 127.5 (12), 125.4 (10/14), 124.6 (2), 76.6 (5), 42.5 (6), 42.1 (7), 40.5 (8), 36.6 (4), 14.8 (6-Me), unit B: 170.2 (1), 154.1 (7), 130.9 (5), 129.5 (4), 128.2 (9), 122.6 (6), 112.4 (8), 56.1 (7-OMe), 54.3 (2), 35.3 (3), unit C: 177.9 (1), 46.5 (3), 42.7 (2), 22.8 (2-Me / 2-Me '), unit D: 170.5 (1), 71.3 (2), 39.5 (3), 24.6 (4), 22.7 (4-Me), 21.3 (5 )

Example 18. The synthesis of cryptophycins 110-112 and 124
Cryptophycin 108
To a mixture of Cryptophycin 90 and Cryptophycin 91 (27 mg, 0.045 mmol) in 0.8 ml of tetrahydrofuran was added 400 μl of an aqueous solution of periodic acid (32 mg, 0.14 mmol). The clear solution was stirred at room temperature for 5 hours. Water was added and the aqueous solution was extracted twice with ethyl acetate. The organic layer was washed with water, dried and concentrated. The residue was purified by reverse phase chromatography on an ODS column (1: 1 MeCN / H ₂ O) to give a Cryptophycin 108% yield of 90%.

¹ Y-NMR (500 MHz) δ-unit A 9.64 (7-Н, d, 1.9), 6.67 (3-Н, ddd, 15.3, 10.0, 5.4), 5.81 (2-Н, dd, 15.3, 0.9) , 5.32 (5-H, ddd, 11.0, 6.6, 2.1), 2.65 (6-H, qdd, 7.2, 6.9, 1.9), 2.53 (4-H, m), 2.44 (4-H ', m), 1.17 (6-Me, d, 7.2); unit B 7.21 (5-H, d, 2.2), 7.08 (9-H, dd, 8.4, 2.2), 6.84 (8-H, d, 8.4), 5.91 (NH, d, 8.4), 4.80 (2- H, m), 3.86 (OMe, S), 3.16 (3-H; dd, 5.4, -14.6), 3.00 (3-H ', dd, 7.8, -14.6); unit C 7.05 (NH, bdd, 6.9, 5.0), 3.47 (3-H, ddd, 4.6, 4.2, -13.5). 3.32 (3-H ', ddd, 6.9, 6.6, -13.5), 2.73 (2-H, m), 1.23 (2-Me; d, 7.2); unit D 4.83 (2-H, dd, 9.5, 3.6), 1.75 (3-H, m), 1.70 (4-H; m), 1.40 (3-H ', ddd, 9.5, 3.9, -14.0), 0.93 (5-H, d, 6.6), 0.88 (4-Me-H, d, 6.6). ¹³ C NMR (125 MHz) δ unit A 200.6 (7), 165.4 (1), 140.5 (3), 125.6 (2), 73.7 (5), 50.1 (6), 36.1 (4), 10.8 (6-Me ), unit B 171.1 (1), 154.0 (7), 131.0 (5), 129.9 (4), 128.3 (9), 122.4 (6), 112.3 (8), 56.1 (OMe), 53.8 (2), 35.0 (3); unit C 175.6 (1), 41.0 (3), 38.1 (2), 14.1 (2-Me); unit D 170.4 (1), 71.3 (2), 39.3 (3), 24.6 (4), 22.9 (5), 21.4 (4-Me).

 Cryptophycin 108 was also obtained by selective ozonolysis of Cryptophycin 82 using the procedure described above for ozonolysis of E in F.

General procedure for the Wittig reaction
Butyllithium (0.4 ml, 2.5 M in hexane) was added to 10 ml of a solution of aryl triphenylphosphonium chloride (1 mmol) in THF at -78 ^° C. The reaction mixture was stirred for 15 minutes at -78 ^° C and then placed for 1 hour in a bath with ice. Three equivalents of the above mixture were added to a solution of Cryptophycin 108 in THF (about 30 mg / ml) at -78 ^° C. The solution was stirred for 20 minutes until the cooling bath was removed. When the temperature rose to 25 ^{° C.} , the reaction was quenched with a saturated aqueous solution of ammonium chloride. The mixture was extracted twice with ethyl acetate. The organic extract was washed with water, dried and concentrated. The residue was purified on an ODS flash column (65:35 MeCN / H ₂ O) to give a mixture of E and Z isomers (the content of 8% to 13% Z isomers depended on the nature of the aryl group; analysis was determined by NMR). The desired E-isomer was crystallized from ether.

Cryptophycin 110
The Wittig reaction led to 5.1 mg of the p-fluorophenyl analogue (containing about 8% of the Z-isomer) from 7.3 mg of Cryptophycin 108 (about 1 mg of unreacted aldehyde was returned). After crystallization from ether, 4.0 mg of pure Cryptophycin 110 was obtained.

[α] _D +42.4 ^o (MeOH, s 2.1). ¹ H NMR δ unit A 7.29 (10-H / 14-H; dd, 8.6, 5.6), 6.98 (11-H / 13-H, dt, 8.6, 8.5), 6.68 (3-H, ddd, 15.3, 9.7, 5.6), 6.38 (8-H, d, 15.8), 5.83 (7-H, dd, 15.8, 8.8), 5.78 (2-H, d, 15.3), 5.00 (5-H, ddd, 10.8, 7.3, 1.3), 2.53 (4-H / 6-H, m), 2.36 (4-H ', m), 1.13 (6-CH ₃ , d, 6.8); unit B 7.21 (5-H, d, 1.8), 7.07 (9-H, dd, 8.4, 1.8), 6.84 (8-H, d, 8.4), 5.68 (NH, d, 8.5), 4.82 (2- H, m), 3.87 (OMe, s), 3.14 (3-H; dd, 5.6, -14.4), 3.04 (3-H ', dd, 7.2, -14.4); unit C, 6.95 (NH, bdd, 6.8, 5.9), 3.50 (3-Н, td, 4.4, -13.5), 3.28 (3-Н ', ddd, 6.8, 6.7, -13.5), 2.72 (2-Н , m), 1.23 (2-Me; d, 7.2); unit D 4.82 (2-H, m), 1.65 (3-H / 4-H; m), 1.35 (3-H ', ddd, 4.5, 3.8, -10.9), 0.78 (5-H, d, 6.4 ), 0.74 (4-Me-H, d, 6.4). ¹³ C NMR (15 MHz) δ unit 165.4 (1), 162.3 (12, d, ¹ J _CF 245.8 Hz), 141.4 (3), 132.9 (9), 130.6 (7), 129.9 (8), 127.6 (10 / 14, d, ³ J _CF 8.0 Hz), 125.2 (2), 115.5 (11/13, d, ² J _CF 21.5 Hz), 77.4 (5), 42.2 (6), 36.4 (4), 17.3 (6 -Me); unit B 170.9 (1), 153.9 (7), 131.0 (5), 129.9 (4), 128.4 (9), 122.4 (6), 112.2 (8), 56.1 (OMe), 53.6 (2), 35.1 (3 ); unit C 175.6 (1), 41.1 (3), 38.3 (2), 14.0 (2-Me); unit D 170.9 (1), 71.5 (2), 39.5 (3), 24.5 (4), 22.7 (5), 21.2 (4-Me).

Cryptophycin 111
The Wittig reaction led to 43 mg of the p-tolyl analogue (containing about 9% Z-isomer) from 55 mg of Cryptophycin 108. After crystallization from ether, 34 mg of pure Cryptophycin 111 was obtained.

[α] _D +44.3 ^o (CHCl ₃ , s 06); EIMS m / z (relative intensity%) 652 (2.9, M ⁺ ), 497 (3.6), 412 (23.8), 242 (20), 145 (46), 105 (75); highresolution EIMS 652.29094 (calcd for C ₃₆ H ₄₅ ClN ₂ O ₇ , Δ-0.6 mmu, M ⁺ ). ¹ N NMR δ unit A 7.21 (10-H / 14-H; d, 8.0), 7.11 (11-H / 13-H, d, 8.0), 6.68 (3-H, ddd, 15.2, 9.6, 5.4) , 6.37 (8 H, d, 15.8), 5.95 (7-H, dd, 15.8, 8.6), 5.76 (2-H, d, 15.2), 4.99 (5-H, dd, 10.4, 6.2), 2.51 ( 4-H / 6-H, m), 2.38 (4-H ', m), 2.32 (12-Me, s), 1.13 (6-CH ₃ , d, 6.8); unit B 7.22 (5-H, d, 2.1), 7.08 (9-H, dd, 8.4, 2.1), 6.83 (8-H, d, 8.4), 5.80 (NH, d, 8.4), 4.81 ( 2-H, m), 3.86 (OMe, s), 3.14 (3-H; dd, 5.6, 14.4). 3.03 (3-H ', dd, 7.3, -14.4); unit C 6.98 (NH, bdd, 6.0, 5.7), 3.49 (3-H, td, 4.6, -13.4), 3.29 (3-H ', ddd, 6.7, 6.6, -13.4), 2.70 (2- H, m) 1.22 (2-Me; d, 7.2); unit D 4.81 (2-H, m), 1.65 (3-H / 4-H; m), 1.37 (3-H ', m), 0.78 (5-H, d, 5.8), 0.73 (4 -Me-H, d, 6.4). ¹³ C NMR (75 MHz) δ unit A 165.5 (1), 141.5 (3), 137.4 (12), 133.9 (9), 131.7 (7), 129.3 (10/14), 129.0 (8), 126.0 (11/13), 125.1 (2), 77.4 (5), 42.2 (6), 36.4 (4), 21.1 (12-Me), 17.3 (6-Me), unit B 171.0 (1), 153.9 (7 ), 131.0 (5), 129.9 (4), 128.4 (9), 122.4 (6), 112.2 (8), 56.1 (OMe), 53.6 (2), 35.1 (3); unit C 175.6 (1), 41.1 (3), 38.3 (2), 14.1 (2-Me); unit D 170.9 (1), 71.6 (2), 39.5 (3), 24.5 (4), 22.7 (5), 21.2 (4-Me).

Cryptophycin 112
The Wittig reaction yielded 35 mg of a 2-thienyl analogue (containing about 13% Z-isomer) from 51 mg of Cryptophycin 108. After crystallization from ether, 25 mg of pure Cryptophycin 111 was obtained.

¹ H NMR (500 MHz) δ unit A 7.12 (12-H; d, 4.9), 6.94 (11-H, dd, 4.9, 3.4), 6.90 (10-H, d, 3.4), 6.68 (3-H , ddd, 15.2, 9.5, 5.3), 6.54 (8-H, d, 15.7), 5.83 (7-H, dd, 15.7, 8.7), 5.78 (2-H, d, 15.2), 4.96 (5 -H, dd, 9.5, 6.5), 2.51 (4-H / 6-H, m), 2.35 (4-H ', m), 1.13 (6-CH ₃ , d, 6.8); unit B 7.21 (5-H, d, 1.6), 7.07 (9-H, dd, 8.4, 1.6), 6.84 (8-H, d, 8.4), 5.74 (NH, d, 7.1), 4.82 (2- H, m), 3.87 (OMe, s), 3.13 (3-H; dd, 5.5, -14.4), 3.04 (3-H ', dd, 7.1, -14.4); unit C, 6.97 (NH, bt, 5.8), 3.50 (3-H, ddd, 4.4, 4.3, -13,4), 3.28 (3-H ', ddd, 6.8, 6.6, -13.4), 2.71 (2 -H, m) 1.22 (2-Me; d, 7.2); unit D 4.82 (2-H, m), 1.67 (3-H / 4-H; m), 1.39 (3-H ', m), 0.80 (5-H, d, 6.4), 0.77 (4-Me -H, d, 6.4).

Cryptophycin 124
The Wittig reaction resulted in 131 mg of the p-chlorophenyl analogue (containing about 10% Z-isomer) from 153 mg of Cryptophycin 108. After crystallization from ether, 107 mg of pure Cryptophycin 124 was obtained;
[α] _D +29.2 ^° (CHCl ₃ , s 0.5); high-resolution EIMS m / z 672.23691 (calcd for С ₃₅ Н ₄₂ Сl ₂ N ₂ O ₇ , Δ 0.0 mmu). ¹ H NMR (500 MHz) δ unit A 7.26 (10-Н / 11-Н / 13-Н / 14-Н, s), 6.68 (3-H, ddd, 15.2, 9.6, 5.4), 6.36 (8- H, d, 15.8), 5.98 (7-H, dd, 15.8, 8.8), 5.77 (2-H, d, 15.2), 5.00 (5-H, bdd, 9.4, 6.3), 2.54 (4-H / 6-H, m), 2.38 (4-H ', m), 1.13 (6-CH ₃ , d, 6.8); unit B 7.22 (5-H, d, 1.7), 7.07 (9-H, dd, 8.4, 1.7), 6.84 (8-H, d, 8.4), 5.73 (NH, bd, 7.8), 4.82 (2- H, m), 3.86 (OMe, s), 3.13 (3-H; dd, 5.5, -14.4), 3.04 (3-H ', dd, 7.2, -14.4); unit C. 6.97 (NH, bt, 5.8), 3.49 (3-H, td, 4.2, -13.4), 3.29 (3-H ', ddd, 6.7, 6.6, -13.4), 2.71 (2-H, m ), 1.22 (2-Me; d, 7.2); unit D 4.81 (2-H, m), 1.65 (3-H / 4-H; m), 1.35 (3-H ', m), 0.77 (5-H, d, 7.1), 0.75 (4-Me -H, d, 7.1). ¹³ C NMR (75 MHz) δ unit A 165.4 (1), 141.3 (3), 135.2 (12), 133.2 (9), 130.9 (7), 130.6 (8), 128.7 (11/13), 127.3 (10 / 14), 125.2 (2), 77.3 (5), 42.2 (6), 36.5 (4), 17.3 (6-Me): unit B 170.9 (1), 153.9 (7), 131.0 (5), 129.8 ( 4), 128.4 (9), 122.4 (6), 112.2 (8), 56.1 (OMe), 53.6 (2), 35.1 (3); unit C 175.6 (1), 41.1 (3), 38.3 (2), 14.1 (2-Me); unit D 170.8 (1), 71.5 (2), 39.6 (3), 24.5 (4), 22.8 (5), 21.3 (4-Me).

Example 19. The synthesis of cryptophycins 115-120 and 126
Cryptophycins 115 and 116
Using the general procedure described above for the epoxidation of styrene-type cryptophycins, Cryptophycin 110 (3.5 mg) was converted to 2.0 mg of Cryptophycin 115 and 1 mg of Cryptophycin 116.

Spectral data for Cryptophycin 115
[α] _D +29.1 ^° (MeOH, s 0.8); EIMS m / z (relative intensity%) 672 (1.9, M ⁺ ), 412 (5.8), 245 (17), 195 (52), 155 (31), 141 (23), 135 (15), 109 (100 ); high-resolution EIMS 668.2853 (calcd for C ₃₅ H ₄₂ ClFN ₂ O ₈ , Δ + 3.4 mmu, M ⁺ ). ¹ N NMR (500 MHz) δ unit A 7.22 (10-H / 14-H; ddt, 8.7, 5.2, 2.0), 7.01 (11-H / 13-H, ddt, 8.7, 8.5, 2.0), 6.68 ( 3-H, ddd, 15.2, 9.7, 5.2), 5.74 (2-H, dd, 15.2, 0.8), 5.15 (5-H, ddd, 11.2, 5.0, 1.8), 3.67 (8-H, d, 2.0 ), 2.88 (7-Н, dd, 7.4, 2.0), 2.54 (4-Н, dtd, 5.2, 1.8, -14.4), 2.44 (4-Н ', ddd, 11.2, 9.7. -14.4), 1.79 ( 6-H, m), 1.13 (6-CH ₃ , d, 6.8); unit B 7.21 (5-H, d, 2.0), 7.06 (9-H, dd, 8.3, 2.0), 6.83 (8-H, d, 8.3), 5.63 (NH, d, 8.4), 4.80 (2- H, ddd, 8.4, 7.2, 5.4), 3.87 (OMe, s), 3.14 (3-H; dd, 5.4, -14.4), 3.03 (3-H ', dd, 7.2, -14.4); unit C 6.94 (NH, bdd, 6.7, 5.0), 3.48 (3-H, ddd, 5.0, 3.7, -13.4), 3.30 (3-H ', ddd, 6.8, 6.7, -13.4), 2.72 (2- H, m), 1.22 (2-Me; d, 7.4); unit D 4.83 (2-H, dd, 9.9, 3.6), 1.70 (3-H / 4-H; m), 1.35 (3-H ', m), 0.87 (5-H, d, 6.5), 0.85 (4-Me-H, d, 6.5). ¹³ C NMR (125 MHz) δ unit A 165.3 (1), 162.9 (12, d, ¹ j _CF 245.4 Hz), 141.0 (3), 132.5 (9), 127.3 (10/14, d, ³ J _CF 8.3 Hz), 125.3 (2), 115.7 (11/13, d, ² J _CF 21.9 Hz), 76.1 (5), 63.0 (7), 58.3 (8), 40.5 (6), 36.7 (4), 13.4 ( 6-Me); unit B 170.9 (1), 154.0 (7), 131.0 (5), 129.7 (4), 128.4 (9), 122.5 (6), 112.3 (8), 56.1 (OMe), 53.6 (2), 35.0 (3 ); unit C 175.6 (1), 41.1 (3), 38.2 (2), 14.1 (2-Me); unit D 170.7 (1), 71.3 (2), 39.4 (3), 24.5 (4), 22.9 (5), 21.3 (4-Me).

Cryptophycins 117 and 118
Using the general procedure described above for the epoxidation of styrene-type cryptophycins, Cryptophycin 111 (6.2 mg) was converted to 3.5 mg of Cryptophycin 117 and 1 mg of Cryptophycin 118.

Spectral data for Cryptophycin 117
[α] _D +25.5 ^° (MeOH, s 1.8); EIMS m / z (relative intensity%) 668 (4.8, M ⁺ ), 412 (6.2), 280 (11), 173 (9.4), 145 (15), 135 (34), 105 (100); high-resolution EIMS m / z 668.28532 (calcd for С ₃₆ H ₄₅ СlN ₂ О ₈ , Δ + 1.1 mmu, M ⁺ ). ¹ H NMR (500 MHz) δ unit A 7.17 / 7.13 (10-Н / 11-Н / 13-Н / 14-Н, A ₂ B ₂ q, 8.0), 6.67 (3-H, ddd, 15.4, 9.8 , 5.6), 5.73 (2-H, dd, 15.4, 0.9), 5.14 (5-H, ddd, 11.2, 4.9, 2.0), 3.65 (8-H, d, 2.0), 2.91 (7-H, dd , 7.6, 2.0), 2.54 (4-Н, bdd, 5.6, -14.3), 2.44 (4-Н ', ddd, 10.7, 9.8, -14.3), 2.35 (12-Ме, s), 1.77 (6- H, m), 1.14 (6-CH ₃ , d, 6.9); unit B 7.21 (5-H, d, 2.2), 7.07 (9-H, dd, 8.5, 2.2), 6.83 (8-H, d, 8.5), 5.65 (NH, d, 8.5), 4.80 (2- H, ddd, 8.3, 7.4, 5.6). 3.87 (OMe, s), 3.13 (3-H; dd, 5.6, -14.5), 3.02 (3-H ', dd, 7.4, -14.5); unit C 6.93 (NH, bd, 6.8, 5.1), 3.48 (3-H, ddd, 5.1, 3.8, -13.2), 3.29 (3-H ', ddd, 6.9, 6.8, -13.2), 2.71 (2- H, m) 1.22 (2-Me: d, 7.1); unit D 4.82 (2-H, dd, 9.8, 3.6), 1.70 (3-H / 4-H; m), 1.33 (3-H ', m), 0.85 (5-H, d, 6.5), 0.84 (4-Me-H, d, 6.5). ¹³ C NMR (125 MHz) δ unit A 165.3 (1), 141.0 (3), 138.4 (12), 133.7 (9), 129.4 (10/14), 125.6 (11/13), 125.3 (2), 76.2 (5), 62.9 (7), 59.0 (8), 40.7 (6), 36.7 (4), 21.1 (12-Me), 13.6 (6-Me); unit B 170.9 (1), 154.0 (7), 131.0 (5), 129.8 (4), 128.4 (9), 122.4 (6), 112.3 (8), 56.1 (OMe), 53.6 (2), 35.1 (3); unit C 175.5 (1), 41.1 (3), 38.3 (2), 14.1 (2-Me); unit D 170.7 (1), 71.3 (2), 39.4 (3), 24.5 (4), 22.8 (5), 21.2 (4-Me).

Spectral data for Cryptophycin 118
^{The 1} H-MR spectrum of the indicated Cryptophycin was similar to the Cryptophycin 38 spectrum with the exception of multiplets at 7.30-7.38 for phenyl protons, replaced by 4H multiplet at 7.15 and 3H singlet at 2.35 for p-tolyl protons.

Cryptophycins 119 and 120
Using the general procedure described above for the epoxidation of styrene-type cryptophycins, Cryptophycin 112 (8 mg) was converted to 1.2 mg of Cryptophycin 119 and 0.5 mg of Cryptophycin 120.

Spectral Data for Cryptophycin 119
¹ H-NMR (500 MHz) δ unit A 7.28 (12-H, d, 5.2), 7.11 (10-H, d, 3.3), 7.00 (11-H, dd, 5.2, 3.3), 6.68 (3- H, ddd, 15.1, 9.9, 5.2), 5.76 (2-H, dd, 15.1, 1.3), 5.15 (5-H, ddd, 11.2, 5.5, 1.8), 3.94 (8-H, d, 2.0), 3.10 (7-H, dd, 7.5, 2.0), 2.56 (4-H, bdd, 5.2, -14.3), 2.44 (4-H ', ddd, 11.2, 9.9, -14.3), 1.76 (6-H, m) 1.13 (6-CH ₃ , d, 6.8); unit B 7.21 (5-H, d, 2.3), 7.07 (9-H, dd, 8.4, 2.3), 6.83 (8-H, d, 8.4), 5.63 (NH, d, 8.4), 4.80 (2- H, ddd, 8.5, 7.4, 5.7), 3.87 (OMe, s), 3.13 (3-H; dd, 5.7, -14.5), 3.03 (3-H ', dd, 7.4, -14.5); unit C 6.95 (NH, bd, 6.5, 5.3), 3.48 (3-H, ddd, 5.3, 3.6, -13.5), 3.31 (3-H ', ddd, 6.8, 6.5, -13.5), 2.72 (2- H, m), 1.23 (2-Me; d, 7.3); unit D 4.85 (2-H, dd, 10.3, 3.5), 1.70 (3-H / 4-H; m), 1.33 (3-H ^' , m), 0.86 (5-H, d, 6.3), 0.85 (4-Me-H, d, 6.3).

Cryptophycins 125 and 126
Using the general procedure described above for the epoxidation of styrene-type cryptophycins, Cryptophycin 124 (47 mg) was converted to 26 mg of Cryptophycin 125 and 12 mg of Cryptophycin 126.

Spectral data for Cryptophycin 125
[α] _D +35.6 ^° (CHCl ₃ , s = 0.9); high-resolution EIMS m / z 688.2301 (calcd for С ₃₅ Н ₄₂ Сl ₂ N ₂ O ₈ , Δ + 1.7 mmu). ¹ H NMR (500 MHz) δ unit A 7.33 (11-H / 13-H, dt, 8.5, 2.0), 7.18 (10-H / 14-H, dt, 8.5. 2.0), 6.67 (3-H, ddd, 15.1, 9.9, 5.2), 5.73 (2-H, dd, 15.1, 0.9), 5.15 (5-H, ddd, 11.0, 4.7, 1.5), 3.66 (8-H, d, 1.9), 2.87 ( 7-H, dd, 7.4, 1.9), 2.53 (4-H, m), 2.42 (4-H ', ddd, 10.6, 10.5, -14.4), 1.78 (6-H, m), 1.12 (6- CH ₃ , d, 7.0); unit B 7.20 (5-H, d, 2.1), 7.06 (9-H, dd, 8.3, 2.1), 6.83 (8-H, d, 8.3), 5.72 (NH, d, 8.1), 4.79 (2- H, ddd, 8.3, 7.9, 5.4), 3.86 (OMe, s), 3.12 (3-H; dd, 5.4, -14.5), 3.02 (3-H ', dd, 7.4, -14.5); unit C 6.97 (NH, bdd, 6.3, 5.5), 3.46 (3-H, ddd, 4.4, 4.1, -13.7), 3.22 (3-H ', ddd, 7.0, 6.3, -13.7), 2.70 (2- H, m), 1.22 (2-Me; d, 7.2); unit D 4.82 (2-H, dd, 9.9, 3.4), 1.70 (3-H / 4-H; m), 1.33 (3-H ', m), 0.87 (5-H, d, 6.5), 0.84 (4-Me-H, d, 6.5). ¹³ C NMR (125 MHz) δ unit A 165.3 (1), 141.0 (3), 135.3 (12), 134.4 (9), 128.9 (11/13), 126.9 (10/14), 125.3 (2), 76.0 (5), 63.1 (7), 58.2 (8), 40.4 (6), 36.7 (4), 13.4 (6-Me); unit B 170.9 (1), 154.0 (7), 131.0 (5), 129.8 (4), 128.3 (9), 122.4 (6), 112.3 (8), 56.1 (OMe), 53.7 (2), 35.0 (3 ); unit C 175.6 (1), 41.0 (3), 38.2 (2), 14.1 (2-Me); unit D 170.7 (1), 71.2 (2), 39.4 (3), 24.5 (4), 22.9 (5), 21.3 (4-Me).

Spectral data for Cryptophycin 126
¹ H-MR (300 MHz) unit A δ 7.26 (11-H / 13-H, d, 8.1), 7.17 (10-H / 14-H, d, 8.1), 6.70 (3-H, ddd, 15.1 , 9.4, 5.3), 5.81 (2-H, d, 15.1), 5.14 (5-H, bdd, 10.0, 4.5), 3.57 (8-H, bs), 2.85 (7-H, bd, 7.6), 2.66 (4-H, m), 2.59 (4-H ', m), 1.77 (6-H, m), 1.04 (6-CH ₃ , d, 7.0); unit B 7.23 (5-H, bs), 7.08 (9-H, bd, 8.4), 6.83 (8-H, d, 8.4), 5.82 (NH, d, 6.8), 4.81 (2-H, ddd, 7.2, 6.8, 5.4), 3.86 (OMe, s), 3.14 (3-H; dd, 5.4, -14.1), 3.03 (3-H ', dd, 7.4, -14.1); unit C 7.03 (NH, bt, 5.7), 3.47 (3-H, ddd, 4.0, 3.7, -13.1), 3.34 (3-H ', ddd, 6.8, 6.4, -13.1), 2.72 (2-H, m) 1.24 (2-Me; d, 7.0); unit D 4.90 (2-H, dd, 10.0, 2.5), 1.74 (3-H / 4-H; m), 1.45 (3-H ', m), 0.91 (5-H, d, 6.5), 0.86 (4-Me-H, d, 6.5). ¹³ C NMR (125 MHz) δ unit A 165.4 (1), 141.3 (3), 135.6 (12), 134.1 (9), 128.8 (11/13), 126.8 (10/14), 125.2 (2), 76.8 (5), 63.3 (7), 55.6 (8), 40.8 (6), 36.7 (4), 13.4 (6-Me); unit B 170.9 (1), 153.9 (7), 131.0 (5), 129.8 (4), 128.4 (9), 122.3 (6), 112.2 (8), 56.1 (OMe), 53.7 (2), 35.0 (3 ); unit C 175.7 (1), 41.0 (3), 38.2 (2), 14.1 (2-Me); unit D 170.8 (1), 71.4 (2), 39.3 (3), 24.6 (4), 23.1 (5), 21.3 (4-Me).

-лейцина (1.245 г, 5 ммолей) в 30 мл безводного CH₂Cl₂ добавляли твердый EDC (1-(3-диметиламинопропил)-3-этилкарбодиимид гидрохлорид) (0.53 г, 2.75 ммоля) в атмосфере N₂ при перемешивании при 0^oС. После перемешивания при 0^oС в течение 1.5 часов, смесь концентрировали ниже 5^oС и остаток разбавляли 35 мл холодного ЕtOАс, последовательно промывали двумя порциями (10 мл) каждого, охлажденного на льду раствора, 5% водного раствора KHSO₄, 5% водного NаНСО₃, рассолом. Органическую фазу отделяли, сушили (MgSO₄) при 5^oС и выпаривали ниже 5^oС. Оставшееся масло разбавляли охлажденным на льду ТГФ (5 мл) и добавляли раствор соединения К (295 мг, 0,5 ммоля) в охлажденный на льду безводный ТГФ (5 мл). Затем были добавлены несколько кристаллов DMAP к смеси, которую перемешивали и позволяли смеси достигнуть 25^oС в течение ночи. Добавляли раствор 5% водного NaHCO₃ (5 мл) и полученную двухфазную смесь интенсивно перемешивали при 25^oС в течение 2 часов. Добавляли ЕtOАс (40 мл), водную фазу экстрагировали добавлением ЕtOАс (2 раза по 20 мл) и объединенные органические слои промывали водой и рассолом, сушили (NaSO₄), фильтровали и выпаривали. Остаток отфильтровывали через слой силикагеля с 25% ЕtOАс в гексане с получением соединения AJ в виде бесцветного масла (385 мг, выход 96%).Example 20. Synthesis of Cryptophycin 121-123 and 127
AJ connection
To a solution of BOC- monohydrate

solid leucine (1.245 g, 5 mmol) in 30 ml of anhydrous CH ₂ Cl _{2 was} added solid EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) (0.53 g, 2.75 mmol) in an atmosphere of N ₂ with stirring at 0 ^o C. After stirring at 0 ^° C for 1.5 hours, the mixture was concentrated below 5 ^° C and the residue was diluted with 35 ml of cold EtOAc, washed successively with two portions (10 ml) of each ice-cooled solution of 5% aqueous KHSO ₄ , 5 % aqueous NaHCO ₃ , brine. The organic phase was separated, dried (MgSO ₄ ) at 5 ^{° C.} and evaporated below 5 ^° C. The remaining oil was diluted with ice-cold THF (5 ml) and a solution of compound K (295 mg, 0.5 mmol) was added to ice-cooled anhydrous THF (5 ml). Then, several DMAP crystals were added to the mixture, which was stirred and allowed to reach 25 ^{° C.} overnight. A solution of 5% aqueous NaHCO ₃ (5 ml) was added and the resulting biphasic mixture was intensively stirred at 25 ^{° C.} for 2 hours. EtOAc (40 ml) was added, the aqueous phase was extracted with EtOAc (2 times 20 ml) and the combined organic layers were washed with water and brine, dried (NaSO ₄ ), filtered and evaporated. The residue was filtered through a layer of silica gel with 25% EtOAc in hexane to give compound AJ as a colorless oil (385 mg, 96% yield).

]_D 13.6^o (с=0.63, CHCCl₃); EIMS m/z (rel. intensity) 805/803/801 (M⁺, 1%), 568/570/572/574 (9/10/6/2), 523/525/527 (5/6/1), 418/420/422/424 (15/15/6/2), 341/343/345/347 (58/70/35/9), 307/309/311 (29/22/10), 224/227/228/229 (10/100/31/14), 208/210/211/2'12 (20/83/45/54); HREIMS m/z 800.2218 (C₃₅H₄₈ ³⁵Cl₄N₂O₈, Δ-5.3 mmu): ¹H NMR (СDСl₃) unit A δ 7.21-7.35 (Ph-H₅; m), 6.77 (3-H; ddd, 6.7/6.7/15.3), 6.4 (8-H; d, 15.8), 6.05 (7-Н; dd, 8.7/15.8), 5.9 (2-Н; d, 15.3), 5.0 (5-Н; m), 2.6 (6-Н; m), 2.55 (4-H₂; m), 1.17 (6-Me; d, 6.7); unit В δ 7.22 (5-H; s), 7.05 (9-H; d, 8.1), 6.84 (8-Н; d, 8.1), 6.55 (NH; d, 7.8), 5.0 (2-Н; ddd, 5.5/7.1/7.8), 4.68-4.8 (CH₂CCl₃; ABq, 11.9), 3.86 (ОМe; s), 3.2 (3-H; dd, 5.5/14.0), 3.06 (3-H'; dd, 7.1/14.0); unit D δ 4.8 (NH; d), 4.2 (2-H; m), 1.65 (4-H; m), 1.55 (3-H; m), 1.4

s), 1.37 (3-H'; m), 0.85 (4-Me; d, 6.5), 0.8 (5-Н; d, 6.5); ¹³C NMR unit А δ 165.4 (1), 139.3 (3), 137.0 (9), 131.5 (8), 130.5 (7), 128.5 (11/13), 127.3 (12), 126.2 (10/12), 125.8 (2), 76.2 (5), 40.8 (6), 33.4 4), 16.8 (6-Me); unit В δ 170.0 (1), 154.2 (7), 131.0 (5), 129.0 (9), 122.5 (6), 112.2 (8), 94.4

74.7

56.1 (ОМe), 53.3 (2), 36.5 (3), unit D δ 173.2 (1), 155.7

80.0

52.4 (2), 41.2 (3), 28.3

24.8 (4), 22.8 (4-Ме), 21.6 (5).[

] _D 13.6 ^o (c = 0.63, CHCCl ₃ ); EIMS m / z (rel. Intensity) 805/803/801 (M ⁺ , 1%), 568/570/572/574 (9/10/6/2), 523/525/527 (5/6/1 ), 418/420/422/424 (15/15/6/2), 341/343/345/347 (58/70/35/9), 307/309/311 (29/22/10), 224 / 227/228/229 (10/100/31/14), 208/210/211 / 2'12 (20/83/45/54); HREIMS m / z 800.2218 (C ₃₅ H ₄₈ ³⁵ Cl ₄ N ₂ O ₈ , Δ-5.3 mmu): ¹ H NMR (CDCl ₃ ) unit A δ 7.21-7.35 (Ph-H ₅ ; m), 6.77 (3- H; ddd, 6.7 / 6.7 / 15.3), 6.4 (8-H; d, 15.8), 6.05 (7-H; dd, 8.7 / 15.8), 5.9 (2-H; d, 15.3), 5.0 (5- H; m), 2.6 (6-H; m), 2.55 (4-H ₂ ; m), 1.17 (6-Me; d, 6.7); unit B δ 7.22 (5-H; s), 7.05 (9-H; d, 8.1), 6.84 (8-H; d, 8.1), 6.55 (NH; d, 7.8), 5.0 (2-H; ddd , 5.5 / 7.1 / 7.8), 4.68-4.8 (CH ₂ CCl ₃ ; ABq, 11.9), 3.86 (OMe; s), 3.2 (3-H; dd, 5.5 / 14.0), 3.06 (3-H '; dd , 7.1 / 14.0); unit D δ 4.8 (NH; d), 4.2 (2-H; m), 1.65 (4-H; m), 1.55 (3-H; m), 1.4

s), 1.37 (3-H '; m), 0.85 (4-Me; d, 6.5), 0.8 (5-H; d, 6.5); ¹³ C NMR unit A δ 165.4 (1), 139.3 (3), 137.0 (9), 131.5 (8), 130.5 (7), 128.5 (11/13), 127.3 (12), 126.2 (10/12), 125.8 (2), 76.2 (5), 40.8 (6), 33.4 4), 16.8 (6-Me); unit B δ 170.0 (1), 154.2 (7), 131.0 (5), 129.0 (9), 122.5 (6), 112.2 (8), 94.4

74.7

56.1 (OMe), 53.3 (2), 36.5 (3), unit D δ 173.2 (1), 155.7

80.0

52.4 (2), 41.2 (3), 28.3

24.8 (4), 22.8 (4-Me), 21.6 (5).

AK connection
Compound AJ (115 mg, 0.134 mmol) was dissolved in TFA (3 ml) and left at 25 ^o C for 1 hour. The solvent was removed and the residue was reevaporated from CH ₂ Cl ₂ to give an amorphous solid. This solid was dissolved in anhydrous THF (5 ml) and a sufficient amount of diethyl isopropylamine was added to adjust the pH of the solution to 8-9 when an aliquot of the solution left a stain on wet pH paper. The solution was cooled to 0 ^{° C.} with stirring, and a solution of compound AL in THF (3 mg) was added. To prepare compound AL, compound Z (55 mg, 0.27 mmol) was dissolved in THF (3 ml) and diethylisopropylamine (0.046 ml, 0.27 mmol) was added. After cooling to -15 ^° C, pivaloyl chloride (0.033 ml, 0.27 mmol) was added dropwise to this mixture, and the solution was stirred at -15 ^° C for 10 minutes and at 0 ^° C for 20 minutes. The resulting suspension was then transferred to a solution of compound AJ in THF. The resulting mixture was stirred at -0 ^{° C.} and allowed to reach a temperature of 25 ^{° C.} overnight. After 12 hours at 25 ^{° C., a} 5% aqueous solution of NaHCO _{3 was} added and the resulting mixture was intensively stirred at 25 ^{° C.} for 1.5 hours. EtOAc (40 ml) was added and the phases were separated. The aqueous phase was extracted additionally with EtOAc (2 times 10 ml). The combined organic extracts were washed with 5% aqueous NaHCO ₃ solution (20 ml), 5% aqueous KHSO ₄ solution (20 ml) and brine, dried (NaSO ₄ ) and evaporated. The remaining oil was chromatographed on silica gel, eluting with 35% EtOAc in hexane to give the AA compound as a colorless foam (98 mg, 83% yield).

3.86 (OMe; s), 3.2 (Н-3; m), 3.1 (Н-3'; dd, 7.2/14.1); unit C δ 5.0 (NH; m), 3.2 (Н2-3; m), 2.55 (Н-2; m), 1.1 (2-Me; d, 7.1); unit D δ 6.12 (NH; m), 4.4 (Н-2; m), 1.65 (H-4; m), 1.55 (Н-3; m), 1.4 (Н-3'; m), 0.86 (4-Me; d, 6.8), 0.81 (5-Н; d, 6.8); ¹³C NMR (CDCl₃) unit A δ 165.8 (1), 138.9 (3), 131.5 (8), 130.4 (7), 128.6 (11/13), 127.4 (12), 126.2 (10/14), 125.8 (2), 76.3 (5), 33.6 (4), 16.6 (6-Me); unit В δ 170.2 (1), 154.1 (7), 136.9 (4), 131.2 (5), 128.6 (9), 122.3 (6), 112.2 (8), 94.4

74.5

56.1 (ОМе), 53.4 (2), 36.6 (3); unit С δ 175.4 (1), 156.3

79.5

43.6 (3), 41.3 (2), 15.1 (2-Me); unit D δ 172.6 (1), 51.3 (2), 40.5 (3), 24.5 (4), 22.7 (4-Ме), 21.5 (5).[α] _D -8.3 (c 0.88, CHCl ₃ ); ¹ H NMR (CDCl ₃ ) unit A δ 7.28-7.35 (Ph-H ₄ ; m), 7.22 (H-12; m), 6.75 (H-3; ddd, 6.4 / 6.4 / 15.4), 6.4 (H- 8; d, 15.9), 6.04 (H-7; dd, 8.6 / 15.9), 5.95 (H-2; d, 15.4), 5.0 (H-5; m), 2.6 (H-4; m), 1.1 (6-Me; d, 6.8); unit B δ 7.22 (Н-5, d, 1.5), 7.15 (NH; d, 7.6), 7.05 (Н-9; dd, 1.5 / 8.2), 6.85 (Н-8; d, 8.2), 5.0 (Н -2; m), 4.8-4.68

3.86 (OMe; s), 3.2 (H-3; m), 3.1 (H-3 '; dd, 7.2 / 14.1); unit C δ 5.0 (NH; m), 3.2 (H2-3; m), 2.55 (H-2; m), 1.1 (2-Me; d, 7.1); unit D δ 6.12 (NH; m), 4.4 (H-2; m), 1.65 (H-4; m), 1.55 (H-3; m), 1.4 (H-3 '; m), 0.86 (4 -Me; d, 6.8), 0.81 (5-H; d, 6.8); ¹³ C NMR (CDCl ₃ ) unit A δ 165.8 (1), 138.9 (3), 131.5 (8), 130.4 (7), 128.6 (11/13), 127.4 (12), 126.2 (10/14), 125.8 (2), 76.3 (5), 33.6 (4), 16.6 (6-Me); unit B δ 170.2 (1), 154.1 (7), 136.9 (4), 131.2 (5), 128.6 (9), 122.3 (6), 112.2 (8), 94.4

74.5

56.1 (OMe), 53.4 (2), 36.6 (3); unit C δ 175.4 (1), 156.3

79.5

43.6 (3), 41.3 (2), 15.1 (2-Me); unit D δ 172.6 (1), 51.3 (2), 40.5 (3), 24.5 (4), 22.7 (4-Me), 21.5 (5).

Cryptophycin 121
Compound AK (73 mg, 0.082 mmol) was dissolved in AcOH (3.5 ml), activated Zn dust was added, and the resulting suspension was subjected to ultrasound for 45 minutes. After stirring at 25 ^{° C. for} an additional 1.5 hours, the mixture was diluted with CH ₂ Cl ₂ (5 ml) and filtered through Celite ^R. The solid was washed additionally with CH ₂ Cl ₂ (10 ml) and the resulting filtrate was evaporated. The residue was dissolved in TFA (3 ml) without further purification, stored at 25 ^{° C.} for 1 hour, and the solvent was removed in vacuo. The residue was re-evaporated from CH ₂ Cl ₂ then from toluene to obtain a colorless solid. This solid was dissolved in DMF (3 ml), FDPP (43 mg, 0.108 mmol) was added, followed by the addition of a sufficient amount of diethylisopropylamine to adjust the pH of the solution to pH 8-9 when an aliquot stained on wet pH paper. After
stirring at 25 ^o C for 16 hours, the mixture was diluted with ether (40 ml) and washed with 5% aqueous KHSO ₄ solution (2 times 1 ml), 5% aqueous NaHCO ₃ solution (15 ml) and brine. After drying (Na ₂ SO ₄ ) and evaporation, the residue was purified by reverse phase gel permeation chromatography on a C-18 silica gel column (Econosil ^R C-18, 22 x 250 mm), eluting with CH ₃ CN / water 65 : 35 at 6 ml / min, the fraction eluted at t _R 48 minutes was collected and evaporated to give an amorphous solid (26 mg, 50% yield).

[α] _D +46.5 ^° (c 0.81, CHCl ₃ ); EIMS m / z 637/639 (M ⁺ , 1%), 449/451 (2/1), 420 (5), 411/413 (7/5), 227/228 (9/7), 195 (10 ), 184 (15), 167/168/169 (40/86/29), 155/157 (100/26), 91 (85), 69 (86); HREIMS m / z 637.2864 (C ₃₅ H ₄₄ ³⁵ ClN ₃ O ₆ , Δ + 5.5 mmu); ¹ H NMR unit A δ 7.21-7.35 (Pn-H ₅ ; m), 6.75 (H-3; ddd, 4.2 / 10.8 / 16), 6.4 (H-8; d, 16), 6.04 (H- 7; dd, 8.8 / 16), 5.75 (H-2; d, 16), 5.1 (H-5; m), 2.55 (H-4; m, H-6; m), 2.35 (H- 4 ';m); unit B δ 7.18 (H-5; d, 2), 7.05 (H-9; dd, 2.0 / 8.3), 6.85 (H-8; d, 8.3), 5.7 (NH; d, 7.2), 4.7 (H -2; ddd, 4.9 / 7.2 / 7.7). 3.86 (OMe; s), 3.1 (H-3; dd, 4.9 / 14.4), 3.0 (H-3 '; dd, 7.7 / 14.4); unit C δ 7.25 (NH; m), 3.5 (H-3; m), 3.4 (H-3 '; m). 2.55 (H-2; m); 1.2 (2-Me; d, 7.2); unit D δ 5.8 (NH; d, 7.2), 4.4 (H-2; m), 1.55 (H-4, m), 1.38 (H ₂ -3; dd, 7.2 / 7.7), 0.76 (4-Me; d, 6.6), 0.74 (H-5; d, 6.6); ¹³ C NMR (CDCl ₃ ) unit A δ 165.1 (1), 141.9 (3), 136.8 (9), 131.7 (8), 130.2 (7), 128.5 (11/13), 127.3 (14), 126.1 (10/12), 125.1 (2), 76.5 (5), 42.3 (6), 36.3 (4), 17.2 (6-Me); unit B δ 171.0 (1), 154.1 (7), 130.8 (5), 129.6 (4), 128.4 (9), 122.5 (6), 112.4 (8), 56.2 (OMe), 54.4 (2), 35.4 (3); unit C δ 175.8 (1), 41.0 (3), 38.6 (2), 14.8 (2-Me); unit D δ 173.2 (1), 51.1 (2), 40.9 (3), 24.7 (4), 23.4 (4-Me), 21.5 (5)
Cryptophycins 122 and 123
To a stirred solution (0 ^° C) of Cryptophycin 121 (0.011 mmol) in anhydrous CH ₂ Cl ₂ (1.3 ml) was added 99% m-CPBA (17 mg, 0.1 mmol) in one portion. Anhydrous toluene (0.7 ml) was then added and the resulting mixture was stirred at 25 ^{° C.} for 72 hours. The solvent was evaporated in vacuo and the remaining solid was purified by reverse phase gel permeation chromatography on a C-18 silica gel column (Econosil ^R C-18, 22 x 250 mm), eluting with a mixture of CH ₃ CM / water 65:35 at 6 ml / min, Cryptophycin 122 (9 mg, 44% yield) was eluted at t _R 37.5 minutes and Cryptophycin 123 (5 mg, 23% yield) was eluted at t _R 40 minutes.

Cryptophycin 122
[α] _D +35.0 (c 1, CHCl ₃ ); EIMS m / z 653/655 (1.6 / 0.7, M ⁺ ), 411/413 (20/5), 280/282 (39/19), 252/254 (13/8), 223/225/227 (19 / 10/23), 211/213 (18/6), 195/197 (51/13), 184/186 (49/11), 176/168/169 (20/16/21), 155/156 / 157 (95/59/42), 139/141/143 (60/40/24), 135/135 (30/11), 129/131 (40/29), 91 (100); HREIMS m / z 653.2906 (C ₃₅ H ₄₄ ³⁵ ClN ₃ O ₇ , Δ-3.8 mmu); ¹ H NMR (d ₆ -acetone) unit A δ 6.65 (H-3; ddd, 38 / 11.0 / 15.0), 5.9 (H-2; dd, 1.9 / 15.0), 5.25 (H-5; ddd, 1.9 / 4.9 / 11.5), 3.82 (H-8; dd, 2.0), 3.0 (H-7; dd, 2.0 / 7.7), 2.65 (H-4; dddd, 2.0 / 2.0 / 3.8 / 14.5), 2.4 ( H-4 '; ddd, 11.0 / 11.5 / 14.5), 1.85 (H-6; dqd, 4.9 / 7.4 / 7.7), 1.1 (6-Me; d, 6.9); unit B δ 7.45 (NH; d, 7.9), 7.22 (H-9; dd, 2.0 / 8.4), 7.0 (H-8; d, 8.4), 4.45 (H-2; ddd, 3.6 / 7.9 / 11.2), 3.84 (OMe; s), 3.2 (H-3; dd, 3.6 / 14.5), 2.75 (H-3 '; dd, 11.2 / 14.5); unit C δ 7.8 (NH; d, 8.8), 3.65 (H-3; ddd, 3.3 / 8.8 / 13.2), 3.1 (H-3 '; ddd, 2.1 / 2.1 / 13.2), 2.55 (H-2; m ); unit D δ 7.35 (NH; d, 8.1), 4.25 (H-2; ddd, 4.8 / 8.1 / 10.8), 1.65 (H-4; m), 1.45 (H-3; ddd, 5.1 / 10.8 / 13.7) , 1.35 (H-3 '; 4.8 / 9.0 / 13.7), 0.8 (4-Me / H-5; d, 6.5); ¹³ C NMR unit A δ 165.9 (1), 140.8 (3), 138.6 (9), 129.4 (11/13), 129.2 (12), 126.7 (2), 126.6 (10/14), 76.0 (5), 63.9 (7), 59.3 (8), 41.2 (6), 37.7 (4), 13.9 (6-Me); unit B δ 171.9 (1), 154.6 (7), 132.5 (4), 131.4 (5), 129.0 (9), 122.4 (6), 113.3 (8), 56.9 (2), 65.4 (OMe), 36.4 ( 3); unit C δ 177.2 (1), 41.3 (3), 38.9 (2), 15.7 (2-Me); unit D δ 174.2 (1), 51.7 (2), 40.6 (3), 25.3 (4), 23.1 (4-Me), 21.5 (5).

Cryptophycin 123
[α] _D +25.2 ^° (c 0.58, CHCl ₃ ); ¹ H NMR (CDCl ₃ ) unit A δ 7.21- 7.35 (Ph-H ₅ ; m), 6.75 (H-3; ddd, 4.1 / 10.9 / 14.1), 5.85 (H-2; dd, 1.4 / 15.2), 5.25 (H-5; m), 3.6 (H-8; d, 2.0), 2.9 (H-7; dd, 2.0 / 7.8), 2.7 (H-4; m), 2.55 (H-4; m) , 1.75 (H-6; m), 1.05 (6-Me; d, 7.1), unit B δ 7.18 (H-5; d, 2.0), 7.05 (H-9; dd, 2.0, 8.5), 6.85 ( H-8; d, 8.5), 5.95 (NH; d, 7.7). 4.7 (H-2; ddd, 4.9 / 7.7 / 8.1), 3.86 (OMe; s), 3.15 (H-3; dd, 4.9 / 14.5), 3.05 (H-3 '; dd, 8.1 / 14.5); unit C δ 7.25 (NH; m), 3.55 (H-3; ddd, 4.6 / 8.7 / 13.3). 3.15 (H-3 '; ddd, 3.0 / 3.1 / 13.3), 2.55 (H-2; m), 1.2 (2-Me; d, 7.3); unit D δ 6.0 (NH; d, 8.2), 4.45 (H-2; m), 1.6 (H-4; m), 1.55 (H ₂ -3; m), 0.88 (4-Me; d, 7.1) 0.87 (5-H; d, 7.1);
Cryptophycin 127
Cryptophycin 122 (5 mg, 0.0075 mmol) was dissolved in CHCl ₃ (2 ml) and cooled to -40 ^o C in an atmosphere of N ₂ . Trimethylsilyl chloride (0.02 ml, 0.157 mmol) was added dropwise and the resulting mixture was stirred at -40 ^° C for 1 hour. The solvent was evaporated and the residue was filtered through a pad of silica gel with 15% EtOH in diethyl ether to give Cryptophycin 127 (4 mg, 77% yield).

[α] _D +28.6 (c 1.16, CHCl ₃ ); EIMS m / z 653 (0.5; M ⁺ -HCl); 411 (2); 182/184 (22/26); 153/155 (68/40); 135 (31); 107/108/109 (55/22/31); 91/92 (100/30); 79/81 (45/35); HREIMS m / z 653, 2841 (C ₃₅ H ₄₄ ³⁵ ClN ₃ O ₇ , Δ 2.6 mmu); ¹ H NMR (CHCl ₃ ) unit A δ 7.3-7.4 (Ph-H5; m), 6.75 (H-3; ddd, 4.2 / 10.9 / 15.0), 5.8 (H-2; dd, 1.5 / 15.0), 5.2 (H-5; ddd, 1.9 / 9.9 / 9.9), 4.65 (H-8; d, 9.6), 4.0 (H-7; brd, 9.6), 2.65 (H-4; m), 2.48 (H-6 ; m), 2.35 (H-4 '; ddd, 10.9 / 11.2 / 14.5), 1.02 (6-Me; d, 7.0); unit B δ 7.18 (H-5; d, 2.2), 7.05 (H-9; dd, 2.2 / 8.6), 6.85 (H-8; d, 8.6), 6.05 (NH; d, 7.7), 4.65 (H -2; ddd, 4.7 / 7.7 / 8.6), 3.86 (OMe; s), 3.15 (Н-3; dd, 4.7 / 14.6), 2.9 (Н-3 '; dd, 8.6 / 14.6); unit C δ 7.25 (NH; brdd, 4.1 / 5.7), 3.4 (H ₂ -3; m), 2.55 (H-2; m), 1.15 (2-Me; d, 7.5); unit D δ 6.2 (NH; d, 8.2), 4.45 (H-2; m), 1.65 (H-4; m), 1.55 (H ₂ -3; m), 0.93 (4-Me; d, 6.8) 0.92 (5-Me; d, 6.8); ¹³ C NMR (CDCl ₃ ) unit A δ 165.4 (1), 142.4 (3), 138.8 (9), 128.8 (11/13), 128.2 (12), 128.0 (10/14), 124.0 (2), 75.6 (5), 73.9 (7), 62.1 (8), 38.6 (6), 36.4 (4), 8.5 (6-Me); unit B δ 171.1 (1), 154.0 (7), 130.8 (5), 129.8 (4), 128.2 (9), 122.4 (6), 112.4 (8), 56.1 (OMe), 54.5 (2), 35.4 ( 3); unit C δ 175.8 (1), 41.1 (3), 38.8 (2), 14.8 (2-Me); unit D δ 173.2 (1), 51.1 (2), 40.9 (3), 25.0 (4), 22.8 (5), 21.8 (4-Me).

Example 21. Synthesis of Cryptophycin 128 and 130-134
Cryptophycins 128, 130 and 131
The crude 2: 1 mixture of 5.7 mg of Cryptophycin 117 and 118 was dissolved in CHCl ₃ (0.5 ml) and treated with trimethylsilyl chloride (5 μl) at -60 ^° C for 3 hours.

 Separation by reverse phase gel permeation liquid chromatography of the resulting chlorohydrin mixture gave Cryptophycin 128, Cryptophycin 130 and Cryptophycin 131. Further purification by normal phase gel permeation liquid chromatography gave pure Cryptophycin 128 (2.1 mg).

Spectral data for Cryptophycin 128
[α] _D +51.4 ^° (CHCl ₃ , s 0.4); IR ν _max 3408, 3281, 2958, 1747, 1731, 1668, 1538, 1505, 1258, 1179, 1067, 910, 733 cm ^-1 ; EIMS m / z (relative intensity%) 668 (0.3. M ⁺ -HCl), 500 (1.2), 445 (4.2), 407 (6), 318 (12), 274 (17), 240 (33), 199 (29), 155 (31), 141 (23), 135 (15), 109 (100); high-resolution EIMS 668.2851 (calcd for C ₃₆ H ₄₅ ClN ₂ O _8. Δ + 1.3 mmu, M ⁺ -HCl). ¹ N NMR (500 MHz) δ unit AA 7.27 (10-H / 14-H, d, 8.0), 7.18 (11-H / 13-H, d, 8.0), 6.69 (3-H, ddd, 15.2, 9.6, 5.4), 5.79 (2-H, dd, 15.2, 0.8), 5.10 (5-H, ddd, 11.0, 8.4, 1.7), 4.63 (8-H, d, 9.6), 3.99 (7-H, bd, 9.6), 2.67 (4-H, bdd, 5.4, -14.2), 2.49 (6-H, dqd, 7.6, 7.6, 1.7), 2.36 (4-H ', td, 10.7, -14.2), 2.35 (12-Me, s), 1.04 (6-CH ₃ , d, 7.1); unit B 7.23 (5-H, d, 2.1). 7.09 (9-H, dd, 8.5, 2.1), 6.84 (8-H, d, 8.5), 5.83 (NH, d, 8.6), 4.80 (2-H, ddd, 8.2, 7.5, 5.7), 3.87 ( OMe, s), 3.16 (3-H; dd, 5.6. -14.4), 3.00 (3-H ', dd, 7.5, -14.4); unit C 6.94 (NH, bt, 5.8), 3.53 (3-H, ddd, 5.4, 4.1, -13.5), 3.24 (3-H ', ddd, 6.9, 6.6, -13.5), 2.74 (2-H, m) 1.22 (2-Me; d, 7.3); unit D 4.92 (2-H, dd, 9.9, 3.2), 1.75 (3-H / 4-H; m), 1.47 (3-H ', m), 0.94 (5-H, d, 6.4), 0.92 (4-Me-H, d, 6.4). ¹³ C NMR (125 MHz) δ unit A 165.6 (1), 141.6 (3), 139.2 (12), 135.3 (9), 129.7 (10/14), 127.9 (11/13), 125.2 (2), 76.4 (5), 74.0 (7), 62.0 (8), 38.4 (6), 36.3 (4), 21.1 (12-Me), 8.6 (6-Me): unit B 171.1 (1), 153.9 (7), 131.0 (5), 130.0 (4), 128.4 (9), 122.4 (6), 112.2 (8), 56.1 (OMe), 53.6 (2), 35.0 (3); unit C 175.3 (1), 41.3 (3), 38.3 (2), 14.0 (2-Me); unit D 170.6 (1), 71.3 (2), 39.7 (3), 24.7 (4), 23.1 (5), 21.5 (4-Me).

Spectral data for Cryptophycin 130
¹ H-NMR (300 MHz) δ unit A 7.27 (10-H / 14-H, d, 7.8), 7.16 (11-H / 13-H, d, 7.8), 6.69 (3-H, ddd, 15.1 , 9.6, 5.2), 5.75 (2-Н, d, 15.1), 5.40 (5-Н, ddd, 10.6, 3.0, 1.8), 5.05 (8-Н, d, 5.9), 3.74 (7-Н, ddd , 10.5, 5.4, 5.2), 2.59 (4-Н, m), 2.55 (6-Н, m), 2.37 (4-Н ', m), 2.33 (12-Me, s), 1.06 (6-СН _3d , 6.9); unit B 7.21 (5-H, bs), 7.07 (9-H, bd, 8.4), 6.83 (8-H, d, 8.4), 5.77 (NH, d, 6.6), 4.80 (2-H, m) 3.87 (OMe, s); 3.12 (3-H; dd, 5.6, -14.4). 3.02 (3-H ', dd, 7.2, -14.4); unit C 7.00 (NH, bt, 6.5), 3.47 (3-H, ddd, 4.3, 4.0, -13.3), 3.24 (3-H ', ddd, 6.7, 6.6, -13.3), 2.72 (2-H, m) 1.23 (2-Me; d, 7.3); unit D 4.80 (2-H, m), 1.70 (3-H / 4-H; m), 1.42 (3-H ', ddd, 7.9, 4.7, -13.0), 0.91 (5-H, d, 6.4 ), 0.86 (4-Me-H, d, 6.4). ¹³ C NMR (125 MHz) unit A 165.5 (1), 142.1 (3), 138.8 (12), 135.2 (9), 129.5 (10/14), 127.5 (11/13), 124.8 (2), 77.8 ( 5), 74.2 (7), 67.2 (8), 39.4 (6), 34.6 (4), 21.1 (12-Me), 12.3 (6-Me); unit B 171.0 (1), 153.9 (7), 131.0 (5), 129.5 (4), 128.4 (9), 112.2 (8), 56.1 (OMe), 53.6 (2), 35.0 (3) 'unit C 175.6 (1), 41.1 (3), 38.3 (2), 14.1 (2-Me); unit D 170.3 (1), 71.5 (2), 39.5 (3), 24.6 (4), 22.7 (5), 21.6 (4-Me).

Spectral data for Cryptophycin 131
¹ H-NMR (300 MHz) δ unit A 7.18 (10-Н / 11-Н / 13-Н / 14-Н, s), 6.64 (3-H, ddd, 15.2, 9.8, 5.4), 5.70 (2 -H, d, 15.2), 5.06 (5-H, bt, 9.1), 4.86 (8-H, d, 9.7), 4.06 (7-H, bd, 9.7), 2.54 (4-H, bdd, 5.2 , -14.2), 2.37 (12-Me, s), 2.13 (4-H ', m), 1.85 (6-H, m), 0.97 (6-CH ₃ , d, 6.6); unit B 7.21 (5-H, d, 2.1), 7.07 (9-H, dd, 8.4, 2.1), 6.83 (8-H, d, 8.4), 5.66 (NH, d, 10.0), 4.79 (2- H, bq, 7.7), 3.87 (OMe, s), 3.14 (3-H; dd, 5.6, -14.4), 3.02 (3-H ', dd, 7.2. -14.4); unit C 6.93 (NH, bt, 6.2), 3.51 (3-H, ddd, 5.0, 4.2. -13.5), 3.27 (3-H ', ddd, 6.7, 6.4, -13.5), 2.73 (2-H, m) 1.23 (2-Me; d, 7.3); unit D 4.86 (2-H, dd, 9.7, 3.4), 1.75 (3-H / 4-H; m), 1.46 (3-H ', m), 0.94 (5-H, d, 6.0), 0.93 (4-Me-H, d, 6.4).

Cryptophycin 132, 133, and 134
A mixture of Cryptophycin 124 and isomer Z (134 mg, 0.199 mmol) was dissolved in dichloromethane (6 ml) and allowed to interact with m-chloroperbenzoic acid (103 mg, 0.598 mmol) at room temperature for 36 hours. Phosphate buffer (10 ml, pH 8) was added to remove the chlorobenzoic acid formed during the reaction. After 30 minutes, the aqueous layer was replaced with dimethyl sulfoxide (50 μl) and a fresh sample of phosphate buffer (10 ml). Stirring was continued for 30 minutes. The organic layer was separated, the solvent was evaporated, and the residue was purified using reverse phase liquid chromatography-gel chromatography (Econosil ODS silica gel, 22 x 250 mm, 35% water / CH ₃ CN, at 6 ml / min) to obtain Cryptophycin 134 (t _R 52.5 min, 9 mg, 6% yield), partially pure Cryptophycin 133 (t _R 61 min, 32 mg, 22% yield) and crude Cryptophycin 132 (t _R 67.5 min, 72 mg). Further purification using normal phase liquid phase gel chromatography (Econosil silica gel, 10 x 250 mm, 56% EtOAc / Hexane, 3 ml / min) gave pure Cryptophycin 132 (65 mg, 45% yield).

Spectral data for Cryptophycin 132
[α] _D +60.1 ^° (CHCl ₃ , c = 1.1); EIMS m / z (relative intensity%) 688 (1.6, M ⁺ -HCl), 412 (10), 261 (11), 195 (57), 184 (28), 165 (28), 155 (92), 135 (85); high-resolution EIMS m / z 688.23563 (calcd for C ₃₅ H ₄₂ Cl ₂ N ₂ O ₈ , Δ -3.8 mmu, M ⁺ -HCl). ¹ H NMR (500 MHz) δ unit A 7.33 (10-H / 11-H / 13-H / 14-H, s), 6.67 (3-H, ddd, 15.2, 9.9, 5.1), 5.79 (2- H, dd, 15.2, 1.1), 5.10 (5-H, ddd, 11.1, 8.1, 1.5), 4.63 (8-H, d, 9.5), 3.98 (7-H, bd, 9.3), 2.62 (4- H, bdd, 5.1, -14.2), 2.47 (6-H, dqd, 7.4, 7.4, 1.5), 2.40 (4-H ', td, 10.8, -14.2), 2.35 (12-Me, s), 1.02 (6-CH ₃ , d, 7.0); unit B 7.21 (5-H, d, 2.1), 7.06 (9-H, dd, 8.4, 2.1), 6.82 (8-H, d, 8.4), 6.04 (NH, d, 8.5), 4.74 (2- H, td, 8.0, 5.4), 3.85 (OMe, s), 3.13 (3-H; dd, 5.3, -14.4), 2.95 (3-H ', dd, 7.8, -14.4); unit C 7.00 (NH, bt, 5.9), 3.48 (3-H, ddd, 5.0. 3.9, -13.4), 3.23 (3-H ', ddd, 6.6, 6.3, -13.4), 2.71 (2-H, m) 1.21 (2-Me; d, 7.2); unit D 4.91 (2-H, dd, 9.7, 3.4), 1.75 (3-H / 4-H; m), 1.45 (3-H ', m), 0.92 (5-H, d, 6.6), 0.91 (4-Me-H, d, 6.6). ¹³ C NMR (125 MHz) δ unit A 165.7 (1), 141.6 (3), 137.3 (12), 134.8 (9), 129.4 (11/13), 129.0 (10/14), 125.2 (2), 76.4 (5), 74.0 (7), 61.4 (8), 38.4 (6), 36.2 (4), 8.7 (6-Me); unit B 171.1 (1), 153.9 (7), 131.0 (5), 130.1 (4), 128.4 (9), 122.3 (6), 112.2 (8), 56.1 (OMe), 53.8 (2), 34.9 (3 ); unit C 175.3 (1), 41.0 (3), 38.1 (2), 14.1 (2-Me); unit D 170.6 (1), 71.3 (2), 39.7 (3), 24.7 (4), 23.1 (5), 21.5 (4-Me).

Spectral data for Cryptophycin 134
^{The 1} H-NMR spectrum is similar to the Cryptophycin 27 spectrum, except that two multiplets at 7.31 (10/14) and 7.36-7.40 (11/12/13) for phenyl protons were replaced by two doublets at 7.26 (10/14) and 7.36 (11/13) for p-chlorophenyl protons.

Example 22. The structure-activity relationship (SAR) and in vivo assessment
The cytotoxicity of Cryptophycins 1, 3, and 8 and new cryptophycins with respect to human KB and LoVo tumor cell lines is shown in Table 6. Cryptophycin 51 and Cryptophycin 3 exhibit comparable cytotoxicity (IC ₅₀ 3.5-5.8 nM). Cryptophycin 52 (IC ₅₀ 43-70 pcM), however, has slightly lower cytotoxicity than Cryptophycin 1 (IC ₅₀ 9-29 pcM), and Cryptophycin 55 (IC ₅₀ 33-47 pcM), has slightly lower cytotoxicity than Cryptophycin 8 ( IC ₅₀ 9-19 pcM). The cytotoxicity of IC ₅₀ for Cryptophycins 117, 122, 125, 127, 128 and 132 is comparable to the cytotoxicity for Cryptophycins 1, 8, 52 and 55; however, the data do not currently allow more complete comparisons.

 Cryptophycin 52 is active in vivo, but requires roughly three times the full dose, compared with Cryptophycin 1. In vivo activity of Cryptophycin 52 against five solid tumors of mouse origin and three human solid tumors is summarized in table 7. Similarly, Cryptophycin 55 was active in vivo, but also required the use of three full dosages, compared with Cryptophycin 8. The in vivo activity of Cryptophycin 55 against six solid tumors of mouse origin and four human solid tumors is summarized in table 8. TED in vivo appears to correlate with the in vitro data.

 Cryptophycins 117, 125, 128 and 132 are active in vivo against pancreatic adenocarcinoma of murine origin (Panc 03). Cryptophycins 117 and 128 are stronger, so they require lower total doses than Cryptophycins 1 and 8, respectively, while Cryptophycins 125 and 132 are less powerful, so they require higher total doses than Cryptophycins 1 and 8, respectively. The data are summarized in table 9.

 T / C values that are less than 42% are considered active according to NCl standards; T / C values that are less than 10% are believed to have superior activity and potential clinical activity according to NCl standards. The total Log kill value is determined as T-C / 3.2 Td, where T represents the average time in days for tumors of the treated group to reach a tumor of 750 mg, C represents the average time in days for tumors of the control group to reach a tumor of 750 mg, and Td represents the doubling time of the tumor volume. T.H. Corbett et. al. , Cytotocxic Anticancer Drugs: Model and Conceptd for Drug Discovery and Development, pp. 35-87; Kluwer: Norwell, 1992). The total values of Log destruction above 2.8, 2.0-2.8, 1.3-1.9, 0.7-1.2 and below 0.7 with a drug treatment duration of 5-20 days are considered ++++, +++, ++, + and - (inactive), respectively . A characterization of activity from +++ to ++++, which is an indication of clinical activity, is necessary for the partial influence or complete regression of masses with 100-300 mg of the size of most transplanted solid mouse tumors. Cryptophycin 52 shows T / C values ranging from 0 to 14% for murine tumors and from 4.1 to 16 for human tumors and the total Log kill values from 1.1 to 2 for murine tumors and from 0 to 3.2 for human tumors. Cryptophycin 1 shows T / C values in the region from 0 to 27% and Log destruction values in the region of less than 1 to 2. Cryptophycin 55 shows T / C values located in the region from more than 0 to 4.4% and Log destruction values in areas from 2.1 to more than 4.6 (curves) for all, but one in three. Colon 26 experiment, which shows a total Log Destruction value of 1.2. Cryptophycin 8 shows T / C values greater than 0 and Log destruction values greater than 2.8 (some curves), as shown in Table 10. It turns out that Cryptophycin 117 and 125 have T / C values and total Log destruction values that are comparable to these for other epoxy type cryptophycins, namely Cryptophycins 1 and 52, while Cryptophycins 128 and 132 have T / C values and total values of Destruction Log, which are comparable with these values for other chlorophycin type cryptophycins, namely Cryptophycin 8 and 55.

 All publications and patent applications listed in this description, but not individually and specifically entered here by reference, are hereby incorporated by reference, as if they were indicated as specifically and individually entered by reference.

 Although the above invention has been described in some detail by way of illustration and example for the purpose of explanation and understanding, it will be apparent to one skilled in the art in light of the description of this invention that some changes and modifications can be made therein without departing from the spirit and scope of the claimed the claims.

Claims (56)

1. The cryptophycin compound represented by the formula:

where Ar represents methyl, or phenyl, or any simple unsubstituted or substituted aromatic or heteroaromatic group;
R ₁ represents halogen;
R ₂ represents OH or
R ₁ and R ₂ may combine to form an epoxy ring, an aziridine ring;
R ₁ and R ₂ may combine to form a double bond between C ₁₈ and C ₁₉ ;
R ₃ represents a lower alkyl group;
R ₄ and R ₅ represent H; or
R ₄ and R ₅ may combine to form a double bond between C ₁₃ and C ₁₄ ;
R ₆ represents a benzyl, hydroxybenzyl (oxybenzyl), alkoxybenzyl, halooxybenzyl, dihalooxybenzyl, haloalkoxybenzyl or dihaloalkoxybenzyl group;
R ₇ , R ₈ , R ₉ and R ₁₀ each independently represent H or a lower alkyl group; and X and Y each independently represent O, NH or alkylamino, provided that the cryptophycin compound is not (see graphic part). 2. The compound of claim 1, wherein R ₈ is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. 3. The compound of claim 1, wherein R ₇ is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. 4. The compound of claim 1, wherein R ₇ is H, R ₈ is methyl, R ₃ is methyl, X and Y are not oxygen. 5. The compound of claim 1, wherein R ₃ is ethyl, propyl, isopropyl, butyl, isobutyl, pentyl or isopentyl. 6. The compound of claim 1, wherein R ₉ is methyl, ethyl, propyl, butyl, isobutyl, pentyl or isopentyl. 7. The compound of claim 1, wherein R ₁₀ is methyl, ethyl, propyl, butyl, isobutyl, pentyl or isopentyl. 8. The compound of claim 1, wherein at least one of the groups attached to C ₃ , C ₆ , C ₁₀ , C ₁₆ , C ₁₇ and C ₁₈ has R stereochemistry. 9. The compound of claim 1, wherein at least one of the groups attached to C ₃ , C ₆ , C ₁₀ , C ₁₆ , C ₁₇ and C ₁₈ has S stereochemistry. 10. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ , R ₇ and R ₈ are methyl, R ₄ and R ₅ are combined together with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents H and X and Y are oxygen. 11. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ , R ₇ and R ₈ are methyl, R ₄ and R ₅ are combined to form a double bonds between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents H and X and Y are oxygen. 12. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ , R ₇ and R ₈ are methyl, R ₄ and R ₅ are combined to form a double bonds between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents H and X and Y are oxygen. 13. The compound of claim 1, wherein Ar is phenyl; R ₁ represents S-chloro; R ₂ represents an R-hydroxyl group; R ₃ , R ₇ and R ₈ are methyl; R ₄ and R ₅ combine to form a double bond between C ₁₃ and C ₁₄ ; R ₆ is 3-chloro-4-methoxybenzyl; R ₉ represents isobutyl; R ₁₀ represents hydrogen; and X and Y are oxygen. 14. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ , R ₇ are methyl, R ₄ and R ₅ are hydrogen, R ₆ is 3- chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents hydrogen, and X and Y are oxygen. 15. The compound of claim 1, wherein Ar is phenyl, R ₁ is S-chloro, R ₂ is R-hydroxyl group, R ₃ , R ₇ and R ₈ are methyl, R ₄ and R ₅ are hydrogen, R ₆ is 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents hydrogen, and X and Y are oxygen. 16. The compound of claim 1, wherein Ar is n-methoxyphenyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 17. The compound of claim 1, wherein Ar is methyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 18. The compound of claim 1, wherein Ar is methyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 19. The compound of claim 1, wherein Ar is methyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 20. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an R, R-aziridine ring, R ₃ , R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₁₀ represents hydrogen, and X and Y are oxygen. 21. The compound of claim 1, wherein Ar is n-fluorophenyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 22. The compound of claim 1, wherein Ar is n-tolyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined together with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 23. The compound of claim 1, wherein Ar is 2-thienyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 24. The compound of claim 1, wherein Ar is n-fluorophenyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 25. The compound of claim 1, wherein Ar is n-fluorophenyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 26. The compound of claim 1, wherein Ar is n-tolyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 27. The compound of claim 1, wherein Ar is n-tolyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 28. The compound of claim 1, wherein Ar is 2-thienyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 29. The compound of claim 1, wherein Ar is 2-thienyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, and X and Y are oxygen. 30. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form the double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X is oxygen, and Y is nitrogen carrying a single hydrogen. 31. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X is oxygen, and Y is nitrogen carrying single hydrogen. 32. The compound of claim 1, wherein Ar is phenyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X is oxygen, and Y is nitrogen carrying single hydrogen. 33. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ and R ₂ are combined to form a double bond between C ₁₈ and C ₁₉ carbon atoms, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined with the formation of a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 34. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ and R ₂ are combined to form an R, R-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X and Y are oxygen. 35. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ and R ₂ are combined to form an S, S-epoxy ring, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X and Y are oxygen. 36. The compound of claim 1, wherein Ar is phenyl, R ₁ is S-chloro, R ₂ is R-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen, X is oxygen, and Y is nitrogen carrying single hydrogen. 37. The compound of claim 1, wherein Ar is n-tolyl, R ₁ is S-chloro, R ₂ is R-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between With ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 38. The compound of claim 1, wherein Ar is n-tolyl, R ₁ is R-chloro, R ₂ is S-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 39. The compound of claim 1, wherein Ar is n-tolyl, R ₁ is R-chloro, R ₂ is an R-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 40. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ is S-chloro, R ₂ is an R-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 41. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ is R-chloro, R ₂ is S-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between With ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen. 42. The compound of claim 1, wherein Ar is n-chlorophenyl, R ₁ is R-chloro, R ₂ is an R-hydroxyl group, R ₃ and R ₇ are methyl, R ₄ and R ₅ are combined to form a double bond between C ₁₃ and C ₁₄ , R ₆ represents 3-chloro-4-methoxybenzyl, R ₉ represents isobutyl, R ₈ and R ₁₀ represent hydrogen and X and Y are oxygen.  43. A pharmaceutical composition for inhibiting the proliferation of a hyperproliferative mammalian cell, characterized in that it comprises an effective amount of a compound of general formula 1 according to claim 1 and a pharmaceutically acceptable carrier.  44. The pharmaceutical composition according to p. 43, characterized in that it further comprises at least one antitumor agent.  45. A method of inhibiting the proliferation of mammalian cells, characterized in that the cells are brought into contact with an effective amount of a compound of general formula 1 according to claim 1.  46. The method according to p. 45, characterized in that the cells are brought into contact with an effective amount of a compound of general formula 1 according to p. 1, additionally containing at least one antitumor agent.  47. The method according to p. 45, wherein the mammalian cell is hyperproliferative.  48. The method according to p. 47, wherein the hyperproliferative cell is human.  49. A method of inhibiting the proliferation of a hyperproliferative mammalian cell having a multiple drug resistance phenotype, characterized in that the cells are contacted with a cryptophycin compound of the general formula 1 according to claim 1 in an amount effective to disrupt the dynamic state of polymerization and depolymerization of a microtubule with blocking cell mitosis.  50. The method according to p. 49, characterized in that the cells are brought into contact with an effective amount of a compound of general formula 1 according to p. 1, additionally containing at least one antitumor agent.  51. The method according to p. 49, characterized in that the hyperproliferative cell is human.  52. A method of alleviating a pathological condition caused by hyperproliferative mammalian cells, comprising administering to the subject an effective amount of a pharmaceutical composition, characterized in that the subject is administered the pharmaceutical composition of Claim 43 to inhibit cell proliferation.  53. The method according to p. 52, wherein the mammalian cells are human.  54. The method according to p. 52, characterized in that it is carried out in combination with another method of the same orientation.  55. The method according to p. 52, characterized in that the pathological condition is characterized by the formation of tumors.  56. The method according to p. 55, characterized in that the pathological condition is caused by tumors of the mammary gland, small chamber lung, non-small chamber lung, calorectal leukemia, melanoma, pancreatic adenocarcinoma, central nervous system (CNS), ovarian prostate, sarcoma of soft tissue or bone, head and neck, gastritis, which includes pancreatic gastritis and gastric gastritis, stomach, myeloma, bladder, renal neuroendocrinia, which includes the thyroid gland and neoplasms not associated with Hodkins disease, and neoplasms associated with Hodkins disease.

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