JPH0114237B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel maytansinoid compound useful as a medicine and a method for producing the same.

More specifically, the invention relates to the formula [In the formula, X is a hydrogen atom or a chlorine atom, R ¹
represents an alkyl group having 2 to 4 carbon atoms, and R ² represents an alkyl group having 2 to 8 carbon atoms which may have a substituent or a methyl group having a substituent. It concerns its manufacturing method.

Regarding the above formula (), the number of carbon atoms represented by R ¹ is 2
-4 alkyl groups include, for example, ethyl, propyl, isopropyl, butyl, sec-butyl,
Mention may be made of the tert-butyl group, of which isopropyl is advantageously used.

Examples of the alkyl group having 2 to 8 carbon atoms represented by R ² include ethyl, propyl, isopropyl, butyl, sec-butyl, bentyl, isopentyl, hexyl, heptyl, and octyl groups.

Examples of substituents on the alkyl group having 2 to 8 carbon atoms or methyl group as R ² include halogen atoms (e.g., chlorine, bromine, iodine), carboxyl groups, alkoxycarbonyl groups having 2 to 5 carbon atoms (e.g., methoxy carbonyl, ethoxycarbonyl,
isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl group), phenoxycarbonyl group, benzyloxycarbonyl group, hydroxyl group, alkoxy group having 1 to 4 carbon atoms (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy , sec-butoxy, tert-
Butoxy groups, etc., and these have 1 to 4 carbon atoms at the end.
alkoxy group or -O-(CH ₂ CH ₂ O) _o -H
(may be replaced with n is an integer from 1 to 5),
Benzyloxy group, alkylthio group having 1 to 4 carbon atoms (e.g., methylthio, ethylthio, propylthio, butylthio group), benzylthio group, phenylthio group, alkylsulfinyl group having 1 to 4 carbon atoms (e.g., methylsulfinyl, ethylsulfinyl,
propylsulfinyl, butylsulfinyl group),
benzylsulfinyl group, phenylsulfinyl group, alkylsulfonyl group having 1 to 4 carbon atoms (e.g.
methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl group), benzylsulfonyl group, phenylsulfonyl group, carbon number 1-
5 alkanoyloxy group (e.g., formyloxy, acetyloxy, propionyloxy, butyryloxy, isobutyryloxy, valeryloxy, pivaloyloxy group), benzoyloxy group,
Phenylacetyloxy group, cyano group, dialkylamino group (e.g. dimethylamino, diethylamino, dibutylamino group), one or two oxo groups (acetalized with lower (C _1-4 ) alcohol, diol, mercaptan, or dimercaptol) or iminated with hydrazine which may have a substituent), a lower (C _{1-4 1-4} ) 1-alkylidene (e.g. methylene, ethylidene, propylidene), phenyl group, α- or β-naphthyl group, vinyl group, ethynyl group, cycloalkyl group having 3-6 carbon atoms (e.g. cyclopropyl, cyclobutyl) , cyclopentyl, cyclohexyl group), 5- to 6-membered ring N,
O, S-containing heterocyclic groups (e.g., pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl,
Piperidinyl, piperazinyl, pyrrolyl, pyrrolidinyl, pyrrolinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, imidazolinyl, pyrazolidinyl, imidazolidinyl, triazolidinyl, furyl, furanyl, tetrahydrofuryl, thienyl, morpholino, oxazolyl, oxazolinyl, thiazolyl, thiazolinyl, oxadiazolyl, thiadiazol (lyl group) and oxiranyl group, dioxolanyl group, and dithiolanyl group.

In addition, each of the above-mentioned cyclic groups, vinyl groups, and ethynyl groups may further have a substituent, such as a C _1-4 alkyl group (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl group), hydroxyl group, C _1-4 alkoxy group (e.g., methoxy,
ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy group), C _1-4 alkanoyloxy group (e.g., formyloxy, acetyloxy, propionyloxy, butyryloxy, isobutyryloxy group),
C _2-5 alkoxycarbonyl group (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl group),
Halogen atoms (e.g. chlorine, fluorine, bromine, iodine),
Nitro group, cyano group, trifluoromethyl group, amino group, mono-C _1-4 -alkylamino group (e.g., methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino group), di-C _1-4 alkylamino group (e.g., dimethylamino, diethylamino, di(2-chloroethyl)amino, dipropylamino, dibutylamino group), C _1-4 alkylthio group (e.g., methylthio,
Ethylthio, propylthio, isopropylthio,
Butylthio, isobutylthio, sec-butylthio,
tert-butylthio group), C _1-4 alkylsulfinyl group (e.g., methylsulfinyl group), C _1-4 alkanesulfonyl group (e.g., methanesulfonyl group),
C _1-4 alkanoylamino group (e.g., formamino,
acetylamino, propionylamino, butyrylamino, isobutyrylamino groups), sulfo groups, sulfamoyl groups (e.g., sulfamoyl, N-methylsulfamoyl, N,N-dimethylsulfamoyl groups), sulfonylamino groups (e.g., methane) Sulfonylamino, benzenesulfonylamino, p-
toluenesulfonylamino), C _1-4 alkanoyl group (e.g., acetyl, propionyl, butyryl, isobutyryl group), benzyloxy group, benzylthio group, benzyloxycarbonyloxy group, tert
-Butoxycarbonyloxy group, benzylamino group, etc.

The compound of the present invention () has the formula It can be produced by alkylating 20-demethoxy-20-hydroxymaytansinol-3-lower carboxylate represented by the formula [wherein X and R ¹ have the same meanings as above].

Alkylation is carried out using an alkylating agent capable of introducing an alkyl group into the 20-position hydroxyl group of the compound (). Such an alkylating agent corresponds to R ² to be introduced.

a Diazo alkanes (e.g., (diazomethane), diazoethane, α-diazotoluene, α-diazoacetophenone, ethyl diazoacetate, diethyl diazomalonate, b trialkyloxonium salts (e.g., (trimethyloxonium fluoroborate), triethyloxonium nium fluoroborate, c halogenide (e.g., (methyl iodide), ethyl iodide, propyl bromide, isopropyl bromide, butyl bromide, pentyl bromide, hexyl bromide, heptyl bromide, octyl iodide, propyl chloride, Butyl chloride, benzyl chloride, benzyl bromide, allyl chloride, allyl bromide, crotonyl bromide, propargyl bromide, bromoacetaldehyde diethylacetal, 3-chloropropionaldehyde
diethyl acetal, chloroacetonitrile,
3-chloropropionitrile, chloroacetone, 4-chloroacetoacetic acid ethyl ester,
1,7-dibromo-2,6-heptadione, 1
-Chloro-3-oxopentane, 1,5-dichloro-2-oxopentane, 1(3)-bromoacetone, α-bromoacetophenone, 2-,3-
or 4-chloro-α-bromoacetophenone, 2,6-dichloro-α-bromoacetophenone, 2-,3- or 4-methyl-α-bromoacetophenone, 2-,3- or 4- Methoxy-α-bromoacetophenone, 2,5-dimethoxy-α-bromoacetophenone, α-chloro-4-fluoroacetophenone, 2- or 4-nitro-α-chloroacetophenone, 4
-Bromo-γ-bromobutylophenone, ethyl chloroacetate, ethyl bromoacetate, ethyl 3-bromopropionate, ethyl bromomalonate, epichlorohydrin,
Epibromohydrin, ethyl bromocyanoacetate, ethyl γ-bromocrotonate,
Chloromethyl methyl ether, 2-bromoethyl ethyl ether Chloromethyl ethyl ether, benzyl chloromethyl ether, chloromethyl methyl sulfide, benzyl chloromethyl sulfide, chloromethyl phenyl sulfide, chloromethyl acetate, chloromethyl pivalate, N-chloromethyl Morpholine, furfuryl chloride, 5-nitrofurfuryl chloride, thenyl chloride, 2-,3
- or 4-picolyl chloride, 5-chloromethyl-2-oxazolidone, 5-chloromethyl-1,2,4-oxadiazole, 1,2-dimethyl-5-chloromethylimidazole, 5-
Methyl-3-chloromethylisoxazole,
5-methyl-2-chloromethylthiazole, 5
-Methylthio-2-chloromethyl-1,3,4
-thiadiazole, 1-methyl-5-chloromethyl-1,2,3-triazole, 1-methyl-5-chloromethyl-tetrazole, 2-chloromethylbenzimidazole, 2-chloromethylbenzo-1,4-dioxane, 5-fluoro-2-bromomethylcoumaran, α-chloromethylnaphthalene, β-bromomethylnaphthalene, d Sulfates: (dimethylsulfate),
Diethyl sulfate, p-toluenesulfonic acid ethyl ester, carbitol p-toluenesulfonic acid ester, hexaethylene glycol monop-toluenesulfonic acid ester, e Isoureas: (O-methyl-), O-ethyl-, O-isopropyl- or O-benzyl-
N,N'-dicyclohexyl isourea, f Quaternary ammonium salts: 1-benzylpyridinium p-toluenesulfonate, 1-{p
-[N,N-bis(2-chloroethyl)amino]benzyl}pyridinium p-toluenesulfonate, g Acetylenes: acetylene dicarboxylic acid ethyl ester, acetylene carboxylic acid methyl ester, cyanoacetylene, etc.

The reaction is preferably carried out in a solvent, such as esters (e.g. ethyl acetate), ethers (e.g. diethyl ether, dioxane,
tetrahydrofuran, etc.) halogenated hydrocarbons (e.g., dichloromethane, chloroform, etc.), nitriles (e.g., acetonitrile, propionitrile, etc.), aromatic hydrocarbons (e.g., benzene, toluene, etc.), pyridine, dimethylformamide, dimethyl sulfoxide, Examples include sulfolane and appropriate mixtures thereof.

The reaction may be carried out at an appropriate temperature, usually within the range of about -20°C to the reflux temperature of the reaction system.

The reaction is usually carried out in the presence of a base. Bases used include alkali metal hydroxides (e.g.
sodium hydroxide, potassium hydroxide, etc.), carbonates (e.g., sodium carbonate, potassium carbonate, etc.),
Tertiary amines (e.g. triethylamine, pyridine,
α-, β- or γ-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-pyrrolidinopyridine, dimethylaniline, diethylaniline, N-methylmorpholine, N-methylpyrrolidine, etc.).

This reaction is a two-phase system consisting of an alkaline aqueous layer and an organic layer, in the presence of a so-called interlayer transfer catalyst (e.g., tetraethylammonium hydroxide, benzyltrimethylammonium bromide, benzyltriethylammonium iodide, cetyltrimethylammonium chloride, cetyltrimethylammonium bromide, etc.). In some cases, it may be preferable to do so. Among the above-mentioned solvents, halogenated hydrocarbons, aromatic hydrocarbon solvents, etc. are particularly preferred as the organic layer in this reaction, and an aqueous solution of an alkali metal hydroxide is particularly preferred as the base (alkaline aqueous layer).

In these alkylation reactions, the alkylating agent may be used in an amount of 1 to 100 molar equivalents, more preferably 1 to 30 molar equivalents, based on the raw material 20-demethoxy-20-hydroxymaytansinoid, and the base may also be used in an amount of 1 to 150 molar equivalents. Molar equivalents, more preferably 1-50 molar equivalents, and interlayer transfer catalysts may be used in 0.1-10 molar equivalents, more preferably 1-5 molar equivalents.

Furthermore, when bromide or chloride is used as an alkylating agent, the reaction time may be shortened by adding an alkali metal iodide (eg, sodium iodide, potassium iodide, etc.) to the reaction mixture. The amount added may be 0.1 to 2 molar equivalents relative to the alkylating agent.

Furthermore, instead of using O-alkylisourea as the alkylating agent, a corresponding alkanol and dicyclohexylcarbodiimide may be used.

In addition, when trying to obtain a target product that has a group that is susceptible to alkylation in the alkyl group (e.g., hydroxyl group, mercapto group, amino group, carboxyl group, etc.), these groups can be It can be obtained by obtaining a protected compound by the method described above and then deprotecting it.

Protecting groups for hydroxyl groups, mercapto groups, and amino groups include lower (C _1-4 ) alkanoyl groups (e.g., formyl group, acetyl group), lower (C _2-5 ) alkoxycarbonyl groups (e.g., methoxycarbonyl group, tert
-butoxycarbonyl group), benzyloxycarbonyl group, halogenated lower (C _1-4 ) alkanoyl group (e.g., trifluoroacetyl, chloroacetyl, dichloroacetyl, bromoacetyl group), etc. are supported as protected carboxyl groups. Lower (C _1-4 ) alkyl ester, benzyl ester, aryl ester (eg, phenyl ester group) groups, etc. are used.

Elimination of these protecting groups is carried out by applying methods known per se (eg reduction, acidolysis, hydrolysis).

Also, R ² is, for example, 5-(pyrazolone-3-yl)
Methyl group, 5-methylpyrazolyl-3-methyl group, 5-(isoxazolon-3-yl)methyl group (in either case, it may be a tautomer formed by migration of a hydrogen atom), or 2 -(isonicotinoylhydrazono)-2-phenylethyl group, etc. () are, for example, each
Hydrazine is a compound in which R ² is 3-methoxycarbonyl-2-oxopropyl, and R ² is 2,4-
Hydrazine to the compound that is dioxopentyl,
^{Hydroxylamine} or
It can also be obtained by reacting a compound in which R ² is phenacyl with isonicotinic acid hydrazide.

The reaction involves alcohol (e.g. methanol, ethanol, isopropanol), tetrahydrofuran,
It is preferable to conduct the reaction in a water-miscible solvent such as dioxane, acetonitrile, dimethylformamide, dimethyl sulfoxide, or a mixed solvent of these and water. Reaction reagents (hydrazine, hydroxylamine, etc.) can be added to the reaction solution as a free base or an appropriate salt, but when using these salts, they should be added in sufficient amounts to form a significant amount of free base in the reaction solution. It is preferable to coexist a base (eg, alkali hydroxide, triethylamine, etc.). These reaction reagents may be used in an amount of 1 to 30 molar equivalents, more preferably 1 to 15 molar equivalents, based on the starting compound (). The reaction can be carried out at any suitable temperature ranging from ice-cooling to the boiling point of the reaction solution.

In addition, for target compounds in which R ² is a group containing a sulfinyl group or a sulfonyl group, a compound in which R ² is a group having a corresponding thioether (thio group) is treated with an appropriate oxidizing agent such as hydrogen peroxide, peracid, etc. (e.g., peracetic acid, pertrifluoroacetic acid, perbenzoic acid, permethchlorobenzoic acid), periodates (e.g.,
It can also be produced by oxidation with sodium salts), permanganates (eg, sodium salts, potassium salts), etc.

Furthermore, when R ² is, for example, a p-aminobenzyl group, it can also be obtained by reducing the corresponding nitro form by means known per se, and this amino form may be an alkylaminosulfonyl form, an alkylamino form, a dialkylamino form, etc. can be derived by means known per se.

The maytansinoid compound () produced by the above alkylation method can be isolated and collected from the reaction mixture using conventional methods such as concentration, solvent extraction, chromatography, recrystallization, etc. .

The maytansinoid compound () of the present invention has strong mitotic inhibitory and antitumor effects and relatively low toxicity.
388, mouse), melanoma (B-16, mouse)]
It has shown a remarkable survival effect when administered to animals suffering from the disease.
It can be used as an effective antitumor agent for cats and humans. Compound () is usually safely administered orally or parenterally as a suitable pharmaceutical composition (eg, injection, etc.) using known carriers, diluents, etc. When the compound () is administered by injection, the route of administration may be appropriately selected from, for example, subcutaneous, intraperitoneal, intravenous, or intramuscular injection, and the dosage is, for example, when used intravenously for melanoma, once from the range of about 1-500 μg/Kg body weight, preferably 5-100 μg/Kg body weight, symptoms,
It can be determined as appropriate in consideration of the target animal, etc.

Injectable solutions can be prepared by conventional methods, such as compound () approx.
It may be prepared by dissolving 50 μg/-about 3 mg in about 0.5 ml of alcohol (eg, ethanol) and adding physiological saline to make a total volume of 10 ml. For smaller doses, this solution can be further diluted with physiological saline.

The maytansinoid compound () of the present invention is also useful in that it exhibits antibacterial activity, such as antifungal and antiprotozoal activity. The compound () can be used advantageously as an antifungal or antiprotozoal agent, for example, when examining the bacterial ecology of soil, activated sludge, or animal body fluids. In other words, when separating useful bacteria from soil, or when testing the effects of protozoa or bacteria other than mold during the operation and analysis of activated sludge methods used in wastewater treatment, mold or bacteria living in the sample may be detected. It is possible to selectively develop bacterial ecology without allowing protozoa to develop. Specifically, a test sample is added to a liquid or solid medium, and 0.1 ml of a 1% methanol-containing aqueous solution containing about 10 to 100 μg/ml of the compound () is added per ml of the medium, followed by culturing.

Maytansinoid compound (2018) can inhibit the growth of pathogenic microorganisms of rice sclerotia, rice leaf blight, and rice sheath blight at 0.02 ml of a 1 mg/ml aqueous solution, so approximately 0.5 -5μg/
A solution of the compound () dissolved in a concentration of ml can be sprayed on rice plants and used to treat these plant diseases.

The raw material compound () used in the production of the compound of the present invention has the formula [In the formula ^, It can be produced by contacting with a culture of a microorganism belonging to the genus Streptomyces or the genus Actinomyces, or a treated product thereof.

In the above, the microorganisms used in the method of converting the methoxy group at position 20 into a hydroxyl group include Bacillus genus, Streptomyces ( Any microorganism belonging to the genus Streptomyces or the genus Actinomyces or its mutant strain may be used. Specific examples of microorganisms that can be used in this method include, for example,
Bacillus megaterium
megaterium) IFO 12108, Streptomyces
Streptomyces flavotricini
IFO 12770, Streptomyces platensis IFO 12901, Streptomyces libani
IFO 13452 and Actinomyces nigrescens IFO 12894. The microorganisms with the above IFO numbers are listed on the List of Cultures by the Fermentation Research Institute.
It is listed in the 1978 edition (Sixth Edition).
The microorganisms listed in this list can be obtained from the Fermentation Research Institute.

In addition, a compound in which X is a hydrogen atom in the above formula (), that is, a dechloromaytansinoid compound, has the formula It is produced by acylating dechloromaytansinol represented by the formula R ¹ -COOH ... () [wherein R ¹ has the same meaning as above] or a reactive derivative thereof, and the formula Dechloromaytansinol () can be produced by reducing a compound of formula () in which X is a chlorine atom using a metal hydride (eg, lithium aluminum hydride).

The present invention will be explained in more detail below using Reference Examples and Examples, but the scope of the present invention is not limited thereto. In addition, in Reference Examples and Examples, unless otherwise specified, Rf values are those measured using silica gel thin layer chromatography (manufactured by Merck & Co., Ltd.).
HPTLC) values are shown. In addition, ansamitocin P-2, P-3, and P-4 are compounds in which X is a chlorine atom and R ¹ is ethyl, isopropyl, and isobutyl, respectively, in the formula (), and PDM-3
is a compound of formula () where X is a chlorine atom and R ¹ is isopropyl, that is, 20-demethoxy-20-
Hydroxymaytansinol 3-isobutyrate, dechloro-PDM-3 is a compound of formula () where X is a hydrogen atom and R ¹ is isopropyl, that is, 19-dechloro-20-demethoxy-20-hydroxymaytansinol 3-isobutyrate. Represent each.

Reference example 1 Antibiotic ansamitocin mixture (Ansamitocin P-2, 12%; Ansamitocin P-3, 71%; Ansamitocin P-
4.17%) was dissolved in 800 ml of dry THF and cooled to -50°C using a dry ice-ethanol bath under a stream of dry nitrogen. Next, 13.0g of lithium aluminum hydride (LAH) was added all at once, stirred at -50℃ to -22℃ for 2 hours, and then heated to -28℃ again.
Cool to -28°C to -22°C and add 3g of LAH.
After stirring at ℃ for 1 hour and 20 minutes, the mixture was cooled again to −50℃, and then 750 ml of 2N hydrochloric acid was carefully added dropwise over the next 30 minutes, and the reaction solution was extracted three times with 2.6, 1.6, and 0.8 portions of ethyl acetate. The extracts were combined, washed with saturated saline (twice to 100 ml), and dried (MgSO ₄ , 250 g). The solvent was distilled off under reduced pressure, and the residue (13.6 g) was subjected to silica gel (1.2 Kg) column chromatography using ethyl acetate/water = 98.5:1.5 (V/
Develop and elute using V), and collect the eluate in fractions of 400 g. Fractions No. 35-52 were combined, the solvent was distilled off, and then dried under vacuum to obtain 7.25 g of maytansinol. Maytansinol and dechloromaytansinol were obtained in the same manner from fractions No. 53-68. Approximately equimolar mixture
1.55 g and 0.78 g of dechloromaytansinol are obtained from fraction No. 69-86. By reprecipitating this product from chloroform-hexane, 0.71 g of dechloromaytansinol is obtained as a white powder.

Melting point: 174-179℃ (decomposition) MS-spectrum (m/e): 469, other UV
-Spectrum (λ ^MeOH _nax ) nm: 231.5, 241.5, 250.5,
277.5, 286 Reference example 2 Dechloromaytansinol 90mg (0.170mmol)
280 mg (1.772 mmol) of isobutyric anhydride and 44 mg of 4-dimethylaminopyridine (DMAP) were dissolved in 10 ml of dry dichloromethane.
(0.361 mmol) and stirred at room temperature for 1.5 hours.
Next, add 22 mg (0.180 mmol) of DMAP and stir at the same temperature for 17 hours. The reaction solution was mixed with 0.5N hydrochloric acid (10ml each twice), sodium bicarbonate solution (10ml), and then water (10ml).
After washing (twice each time), drying and distilling off the solvent. Dissolve the residue (174 mg) in chloroform,
Silica gel column chromatography (column 20mm (outer diameter)
×400mm) and chloroform/ethanol =
Elutes from 100/1 to 40/1. The eluate was fractionated into 25 g portions, fractions Nos. 42-65 were combined, and the solvent was distilled off to obtain 69 mg of crude dechloromaytansinol 3-isobutyrate. Dissolve this substance in ethyl acetate, leave it to stand, and remove the precipitated crystals.
mg of dechloromaytansinol 3-isobutyrate is obtained as white prismatic crystals.

Melting point: 250-252℃ (decomposition) MS-spectrum (m/e): 600, 557, 539,
524, other UV-spectrum (λ ^MeOH _nax ) nm: 232.5, 241,
251, 277.5, 285.5 Reference example 3 Streptomyces flavotrichini IFO 12770
, dextrin 1%, glucose 1%, glycerol 1%, peptone 0.5%, yeast extract 0.5%, meat extract 0.5%, salt 0.3% and calcium carbonate 0.5
% in a medium (PH7.2) and culture with shaking at 28°C for 48 hours. 20 mg of ansamitocin P-3 is added to this culture solution 2, shaken at 28°C for 48 hours to react, and then extracted with ethyl acetate. The extract is filtered, washed with dilute hydrochloric acid, aqueous sodium bicarbonate, and water, dried, and concentrated under reduced pressure. Add petroleum ether to the residue, dissolve the precipitate in a small amount of chloroform,
Purification by silica gel chromatography yields 12 mg of white powder of PDM-3. Melting point: 165-168℃ Reference Example 4 Dechloromaytansinol 3-isobutyrate was converted to dechloro-
PDM-3 is obtained. Rf=0.42 [Developing solvent, chloroform:methanol=9:1, plate: silica gel glass plate (Merck 60F ₂₅₄ )] Example 1 Dissolve 31 mg of PDM-3 in 1.0 ml of dichloromethane, add 1.0 ml of water and 1N-sodium hydroxide. aqueous solution
After adding 50 μl, 16 mg of cetyltrimethylammonium chloride and ethyl p-toluenesulfonate.
Add 15mg. The reaction mixture was vigorously stirred at room temperature for 20 hours, then acidified with 100 μl of 1N hydrochloric acid and extracted with chloroform. The solvent is distilled off under reduced pressure and the residue is separated and purified by silica gel column chromatography to obtain 4.8 mg of PDM-3-C ₂₀ -ethyl ether. Rf = 0.44 (developing solvent, chloroform:methanol = 95:5), MS-spectrum (m/e), 648 (M ⁺ ) 587 (M ⁺ -61) Example 2 Dissolve 62 mg of PDM-3 in 4.0 ml of tetrahydrofuran Then, add 0.2 ml of 1N caustic soda aqueous solution and cool to -10 to -5°C. Add to this solution under stirring,
Triethyloxonium borofluoride 47.5
mg and react at -5 to 0°C for 15 minutes. After adding 20 ml of tetrahydrofuran to the reaction solution and drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was separated and purified by silica gel column chromatography to obtain 58 mg of PDM-3-C ₂₀ -ethyl ether. It will be done.

Rf value and MS-spectrum data are from Example 1
Same as .

Example 3 Dissolve 99.2 mg of PDM-3 in 3.2 ml of dichloromethane, add 3.2 ml of water and 0.2 ml of 1N aqueous sodium hydroxide solution.
ml, then 51 mg of cetyltrimethylammonium chloride and 80 mg of ethyl bromoacetate are added and the reaction mixture is stirred vigorously for 1.5 hours at room temperature.
After neutralization with 0.24 ml of 1N hydrochloric acid, extraction with chloroform, the solvent was distilled off under reduced pressure, and the residue was separated and purified by silica gel column chromatography, resulting in 70 mg of
PDM-3-C ₂₀ -ethoxycarbonyl methyl ether is obtained. Rf = 0.54 (developing solvent, chloroform:methanol = 95.5), MS-spectrum (m/e): 706 (M ⁺ ), 688 (M ⁺ -18), 663 (M ⁺ -
43), 645 (M ⁺ -61) Example 4 In the same manner as in Example 3, 31 mg of PDM-3 was prepared from 124 mg and 32 mg of isopropyl bromoacetate.
PDM-3-C ₂₀ -isopropyloxycarbonyl methyl ether is obtained. MS-spectrum (m/e): 720 (M ⁺ ), 677 (M ⁺ -43), 659 (M ⁺ -
61) Example 5 In the same manner as in Example 3, PDM-3, 124 mg and
tert-butyl bromoacetate 117mg to 121mg
PDM-3-C ₂₀ -tertbutoxycarbonyl methyl ether is obtained. MS-spectrum (m/
e): 673 (M ⁺ -61) Example 6 In the same manner as in Example 3, 128 mg of PDM-3, 124 mg and 86 mg of phenyl bromoacetate was prepared.
PDM-3-C ₂₀ -phenoxycarbonyl methyl ether is obtained. MS-spectrum (m/
e): 693 (M ⁺ -61) Example 7 In the same manner as in Example 3, 68.5 mg of PDM-3 was prepared from 62 mg of PDM-3 and 40 mg of phenacyl bromide.
-C ₂₀ - phenacyl ether is obtained.

MS-spectrum (m/e): 738 (M ⁺ ), 777
(M ⁺ -61) Example 8 In the same manner as in Example 3, 43 mg of PDM-3- was prepared from 174 mg of PDM-3 and 165 mg of bromoacetal.
C ₂₀ -β,β-diethoxyethyl ether is obtained. MS-spectrum (m/e): 736 (M ⁺ ),
708 (M ⁺ -28), 675 (M ⁺ -61) Example 9 In the same manner as in Example 3, 84 mg was prepared from 124 mg of PDM-3 and 84 mg of ethyl γ-bromoacetoacetate.
PDM-3-C ₂₀ -γ-ethoxycarbonylacetonyl ether is obtained. MS-spectrum (m/e): 687 (M ⁺ -61) Example 10 In the same manner as in Example 3, 64 mg of PDM-3-C ₂₀ was obtained from 62 mg of PDM-3 and 34 mg of benzyl bromide.
- Benzyl ether is obtained. MS-spectrum (m/e): 710 (M ⁺ ), 667 (M ⁺ -43), 649 (M ⁺
-61) Example 11 In the same manner as in Example 3, 53 mg of PDM-3-C ₂₀ - was prepared from 124 mg of PDM-3 and 56 mg of chloroacetone.
Acetonyl ether is obtained.

MS-spectrum (m/e): 676 (M ⁺ ), 633
(M ⁺ −43), 615 (M ⁺ −61) Example 12 76 mg of PDM-3-C ₂₀ -phenoxycarbonyl methyl ether obtained in Example 6 was mixed with methanol 10
ml, and add 0.1 ml of 1N aqueous sodium hydroxide solution while stirring at room temperature. After 30 minutes, add 0.1 ml again, and after another 30 minutes, add 0.1 ml (total 0.3 ml), and after adding the entire amount, leave it for 15 minutes. Add 1N to this solution
After neutralizing by adding 0.3 ml of hydrochloric acid, the mixture is concentrated to dryness under reduced pressure, and the residue is separated and purified by silica gel chromatography to obtain 68 mg of PDM-3-C ₂₀ -carboxymethyl ether. Rf=0.31 (developing solvent, acetonitrile:water 95:5) MS-spectrum (m/e): 617 (M ⁺ -61) Example 13 PDM-3-C ₂₀ -γ-ethoxy obtained in Example 9 Dissolve 38 mg of carbonylacetonyl ether in 1.0 ml of methanol, and add 5 mg of hydratine hydrate while stirring at room temperature. After 30 minutes, 5 mg of hydratine hydrate is added and left for 1 hour. The solvent was dried under reduced pressure and the residue was separated and purified by silica gel column chromatography to obtain 18 mg of PDM-3-
C ₂₀ -(5-pyrazolon-3-yl)methyl ether is obtained. Rf=0.11 (developing solvent, chloroform:methanol 95:5) MS-spectrum (m/e): 655 (M ⁺ -61) Example 14 In the same manner as in Example 3, 124 mg of PDM-3 and chloromethyl pivalate, 1120mg more than 60mg
PDM-3-C ₂₀ -pivaloyloxymethyl ether is obtained. MS-spectrum (m/e): 673
(M ⁺ −61) Example 15 Dechloro-PDM-359.2 in the same manner as Example 2
mg, 0.2 ml of 1N caustic soda aqueous solution, and 51 mg to 53 mg of triethyloxonium borofluoride.
Dechloro-PDM-3-C ₂₀ -ethyl ether is obtained. MS-spectrum (m/e): 614
(M ⁺ ), 553 (M ⁺ −61) Example 16 PDM-3 105.8 mg, allyl bromide 123 mg, and cetyltrimethylammonium bromide 124 mg
Dissolve in 5 ml of dry dichloromethane and add
Add 10.2 ml of 0.1N sodium hydroxide aqueous solution and stir at room temperature for 2 hours. After neutralizing with 1N hydrochloric acid,
Add saturated brine, separate the organic layer, wash with brine, dry (Na ₂ SO ₄ ), and then evaporate the solvent. The residue was dissolved in ethyl acetate, the insoluble matter was removed, the liquid was concentrated, the residue was chromatographed on silica gel (approximately 40 g of SiO ₂ , solvent: ethyl acetate (approximately 50 ml), and ethyl acetate/water saturated ethyl acetate = 4/ 1
(V/V) and collect the eluate in fractions of 17 g each.
Fractions No. 9-16 were combined, the solvent was distilled off, and 50.5 mg of
PDM-3-C ₂₀ - Obtain allyl ether. MS―
Spectrum (m/e): 565 (M ⁺ -61) Example 17 In the same manner as in Example 16, 106 mg of PDM-3, 122 mg of propargyl bromide, 10.3 ml of 0.1N NaOH,
Cetyltrimethylammonium bromide 124.5mg
was reacted at room temperature for 3 hours, and the reaction solution was worked up and chromatographed in the same manner as in Example 16 to obtain 41.7 mg of
PDM-3-C ₂₀ - Obtain propargyl ether.
MS-spectrum (m/e): 563 (M ⁺ -61) Example 18 In the same manner as in Example 16, 94.2 mg of PDM-3,
50μl of chloroacetonitrile, 6.1ml of 0.1N NaOH,
and cetyltrimethylammonium bromide
Stir 110.4 mg at room temperature overnight. Further, 25 μl of chloroacetonitrile and 4.0 ml of 0.1N NaOH were added, and the mixture was stirred for a further 7 hours, and then the reaction solution was worked up according to Example 16. The residue was subjected to silica gel chromatography (SiO ₂ , 44 g; solvent: chloroform/methanol = 20/1 (V/V); fractionated into 20 g portions),
34.3 mg of PDM-3-C ₂₀ -cyanomethyl ether is obtained from fractions No. 21-25. MS-spectrum (m/
e): 564 (M ⁺ -61) Example 19 Add chloromethyl methyl sulfide to a solution of 82.8 mg of PDM-3 dissolved in 3 ml of dry dichloromethane.
Add 100 μl and 100 μl of triethylamine and stir at room temperature for 5 hours. The solvent was distilled off, the residue was dissolved in ethyl acetate, washed with brine and dried, and the solvent was distilled off. The residue was chromatographed on silica gel according to Example 3 (solvent: ethyl acetate), and 25.3 mg of PDM was extracted.
-3-C ₂₀ -methylthiomethyl ether is obtained.
MS-spectrum (m/e): 585 (M ⁺ -61) Example 20 In the same manner as in Example 3, 72 mg of PDM-3, 124 mg and 72 mg of 3-bromoacetylacetone was
PDM-3-C ₂₀ -(diacetyl)methyl ether is obtained. MS-spectrum (m/e): 657
(M ⁺ -61) Example 21 In the same manner as in Example 3, 56 mg of PDM-3, 128 mg and 286 mg of 1-bromoacetylacetone was prepared.
PDM-3-C ₂₀ -(γ-acetyl)acetonyl ether is obtained. MS-spectrum (m/
e): 657 (M ⁺ -61) Example 22 In the same manner as in Example 3, 46 mg of PDM-3 and 161 mg of p-chlorobenzyl chloride were prepared.
PDM-3-C ₂₀ (p-chloro)benzyl ether is obtained. MS-spectrum (m/e): 744
(M ⁺ ), 683 (M ⁺ -61) Example 23 In the same manner as in Example 3, 142 mg of PDM-3, 124 mg and 234 mg of p-chlorophenacyl bromide was prepared.
PDM-3-C ₂₀ -(p-chloro)phenacyl ether is obtained. MS-spectrum (m/e):
772 (M ⁺ ), 711 (M ⁺ -61) Example 24 In the same manner as in Example 3, 281 mg of PDM-3 and 432 mg of p-nitrobenzyl bromide were prepared.
PDM-3-C ₂₀ -(p-nitro)benzyl ether is obtained. MS-spectrum (m/e): 694
(M ⁺ -61) Example 25 In the same manner as in Example 3, 124 mg of PDM-3 and 217 mg of 3,4,5-trimethoxybenzyl chloride
57 mg of PDM-3-C ₂₀ -(3,4,5-trimethoxy)-benzyl ether is obtained.
MS-spectrum (m/e): 739 (M ⁺ -61) Example 26 In the same manner as in Example 3, from 124 mg of PDM-3 and 187 mg of 3,4-dimethoxybenzyl chloride.
13mg PDM-3- _C20- (3,4-dimethoxy)
Benzyl ether is obtained. MS-spectrum (m/e): 709 (M ⁺ -61) Example 27 In the same manner as in Example 3, 127 mg of PDM-3 and 196 mg of p-cyanobenzyl bromide were obtained.
PDM-3-C ₂₀ -(p-cyano)benzyl ether is obtained. MS-spectrum (m/e): 674
(M ⁺ -61) Example 28 In the same manner as in Example 3, 62 mg was prepared from 124 mg of PDM-3 and 229 mg of p-acetoxybenzyl bromide.
PDM-3-C ₂₀ -(p-acetoxy)benzyl ether is obtained. MS-spectrum (m/
e): 768 (M ⁺ ), 707 (M ⁺ -61) Example 29 In the same manner as in Example 1, PDM-3,124 mg and carbitol p-toluenesulfonic acid ester
18 mg of PDM-3-C ₂₀ -(2-ethoxy)ethoxyethyl ether is obtained from 288 mg.

MS-spectrum (m/e): 736 (M ⁺ ), 675
(M ⁺ -61) Example 30 In the same manner as in Example 3, 12 mg of PDM-3-C ₂₀ -(7-bromo- 2,6
-dioxo)heptyl ether is obtained. M.S.
- Spectrum (m/e): 764 (M ⁺ -61) Example 31 In the same manner as in Example 1, 23 mg of PDM-3- was obtained from 124 mg of PDM-3 and 436 mg of hexaethylene glycol monop-toluenesulfonic acid ester. _C20
-Hexaethylene glycolyl ether is obtained. MS-spectrum (m/e): 823 (M ⁺ -61) Example 32 151 mg of PDM-3-C ₂₀ -(p-nitro)benzyl ether obtained in Example 24 was mixed with methanol 12
ml, add 3 ml of water, then add calcium chloride.
Add a solution of 45 mg dissolved in 0.5 ml of water. This was reacted with 150 mg of zinc powder at 60°C for 1 hour. After cooling, insoluble matter was removed, the liquid was concentrated, the residue was extracted with chloroform, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure, yielding 141 mg of PDM-3-C ₂₀ -.
(p-amino)benzyl ether is obtained.
MS-spectrum (m/e): 664 (M ⁺ -61), 559
(M ⁺ -166) Example 33 36.3 mg of PDM-3-C ₂₀ -(p-amino)benzyl ether obtained in Example 32 was dissolved in 5.0 ml of dry dichloromethane, and the solution was dissolved in dry pyridine while stirring at room temperature.
Add 160 mg and 115 mg of methanesulfonic acid chloride. After standing for 3 hours, add 15 ml of chloroform and 15 ml of water and separate the organic layer. The aqueous layer is further chloroformed.
Extract twice with 15 ml. After drying the organic layer over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure and the residue was separated and purified by silica gel chromatography to yield 27 mg.
PDM-3-C ₂₀ -(p-methanesulfamide)benzyl ether is obtained. MS-spectrum (m/e): 742 (M ⁺ -61) Example 34 PDM-3-C ₂₀ -(p-acetoxy)benzyl ether obtained in Example 28 27mg methanol
Dissolve in 5.4 ml and add 0.54 ml of triethylamine while stirring at room temperature. After 3 hours, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 23 mg of PDM-3-C ₂₀ -(p-
hydroxy)benzyl ether is obtained. M.S.
- Spectrum (m/e): 665 (M ⁺ -61) Example 35 171 mg of PDM-3 was dissolved in 5 ml of dichloromethane, 70 μl of acetylene carboxylic acid methyl ester and 15 μl of N-methylmorpholine were added, and the mixture was dissolved at room temperature for about 3 Stir for an hour. Wash the reaction solution with 0.1N hydrochloric acid and separate the organic layer. The aqueous layer is extracted with chloroform, and the organic layers are combined, washed with water, and dried with Glauber's salt.
After removing Glauber's salt and distilling off the solvent, the residue is separated and purified by silica gel column chromatography. The fraction that elutes first is called fraction-1, and 48 mg of
PDM-3-C ₂₀ -E-methoxycarbonyl vinyl ether, the latter fraction was designated as fraction-2, 27mg
PDM-3-C ₂₀ -Z-methoxycarbonyl vinyl ether is obtained. Fraction 1 (E form): Rf=0.76
(Developing solvent, water-saturated ethyl acetate) NMR (90MHz,
CDCl ₃ ), δ7.75 (d, J = 12, 1H), 5.58 (d, J
= 12, 1H); MS-spectrum (m/e) 704
(M ⁺ ), 643 (M ⁺ −61). Fraction 2 (Z form): Rf=0.70
(Developing solvent, water-saturated ethyl acetate); NMR (90MHz,
CDCl ₃ ) δ6.77 (d, J = 6, 1H), 5.30 (d, J =
6,1H); MS-spectrum (m/e) 704 (M ⁺ ),
643 (M ⁺ −61).

Example 36 Dissolve 31 mg of PDM-3 in 3 ml of dichloromethane, and add 30 μl of acetylene dicarboxylic acid ethyl ester.
After adding 15 mg of 4-dimethylaminopyridine, the mixture was stirred at room temperature overnight. Thereafter, the reaction solution is concentrated and the residue is separated and purified by silica gel column chromatography. Fraction-1 in the same manner as Example 35
(3.4 mg) and fraction-2 (2 mg) are obtained. these are
PDM-3-C ₂₀ is a geometric isomer of 1',2'-diethoxycarbonyl vinyl ether. Fraction 1: Rf = 0.83 (developing solvent, water-saturated ethyl acetate), MS-spectrum (m/e) 729 (M ⁺ -
61), Fraction 2: Rf = 0.76 (developing solvent, water-saturated ethyl acetate) MS-spectrum (m/e) 729 (M ⁺ -
61).

Example 37 To a solution of 97 mg of PDM-3 dissolved in 5 ml of dichloromethane, add 32 μl of cyanoacetylene and 5 μl of N.
- Add methylmorpholine and stir at room temperature for 30 minutes. The reaction solution was dried under reduced pressure, and the residue was dissolved in ethyl acetate and washed with 0.5N hydrochloric acid and then with brine.
The solvent was distilled off, and the residue was chromatographed on silica gel (40 g) (solvent: ethyl acetate/water saturated ethyl acetate =
3/1 (V/V)), and the eluate is fractionated into 17 g portions and collected. Fraction Nos. 9-11 were combined and the solvent was distilled off, and 39 mg of PDM-3-C ₂₀ -E-β-cyanovinyl ether (compound A) was obtained from fractions No. 14-25 in the same manner. 37 mg of 3-C ₂₀ -Z-β-cyanovinyl ether (compound B) is obtained.

Compound A: MS-spectrum (m/e): 610
(M ⁺ −61) NMR (in CDCl ₃ ) δ: 5.14 and 7.46 (respectively
1H, d, J = 13Hz), others Compound B: MS-spectrum (m/e): 610
(M ⁺ −61) NMR (in CDCl ₃ ) δ: 5.25 and 7.61 (respectively
1H, d, J = 7 Hz), Other Examples 38 In the same manner as in Example 3, 87 mg of PDM-3 and 177 mg of α-chloromethylnaphthalene were prepared.
PDM-3-C ₂₀ -(α-naphthyl)methyl ether is obtained. MS-spectrum (m/e): 760
(M ⁺ ), 699 (M ⁺ −61).

Example 39 In the same manner as in Example 3, 116 mg of PDM-3 was prepared from 124 mg of PDM-3 and 221 mg of β-bromomethylnaphthalene.
PDM-3-C ₂₀ -(β-naphthyl)methyl ether is obtained. MS-spectrum (m/e): 760
(M ⁺ ), 699 (M ⁺ −61).

Example 40 20 mg of PDM-3-C ₂₀ -phenacyl ether obtained in Example 7 was dissolved in 2.7 ml of methanol,
This includes 37mg isonicotinic acid hydrazide and 22mg
After adding sodium acetate, the mixture was stirred and reacted at 60°C for 3 days. The solvent was distilled off under reduced pressure, and the chloroform-soluble portion was separated and purified using silica gel (10 g) chromatography (solvent: methanol/chloroform = 5/95 (V/V)) to obtain 8 mg of PDM-3-C ₂₀ -(2- Isonicotinoylhydrazono-2-phenylethyl ether is obtained. MS-spectrum (m/
e): 857 (M ⁺ ).

Example 41 In the same manner as in Example 1, PDM-3, 124 mg and
From 192 mg of 1-{p-[N,N-bis(2-chloroethyl)amino]benzyl}pyridinium p-toluenesulfonate, 38 mg of PDM-3-C ₂₀ -
p-[N,N-bis(2-chloroethyl)amino]benzyl ether is obtained. MS-spectrum (m/e): 849 (M ⁺ ), 788 (M ⁺ -61).

Claims (1)

[Claims] 1 formula [In the formula, X is a hydrogen atom or a chlorine atom, R ¹
represents an alkyl group having 2 to 4 carbon atoms, and R ² represents an alkyl group having 2 to 8 carbon atoms which may have a substituent or a methyl group having a substituent. 2 formulas [In the formula, X is a hydrogen atom or a chlorine atom, R ¹
represents an alkyl group having 2 to 4 carbon atoms] A maytansinoid represented by [wherein X and R ¹ have the same meanings as above, and R ² represents an alkyl group having 2 to 8 carbon atoms which may have a substituent or a methyl group having a substituent] Method of manufacturing compounds.