JPH066593B2 - Mitomycin derivative - Google Patents

Description

TECHNICAL FIELD The present invention relates to a novel mitomycin derivative having antitumor activity.

Prior Art and Problems Mitomycins are generally known as antibiotics having antibacterial activity and antitumor activity. Typical mitomycins include mitomycins A, B, C and porphyromycin (Merck index 10th edition), mitomycins D and E (JP-A-54-122797), mitomycins F and J (JP-A-55-45322), and the like. Can be given. These mitomycins have the chemical structures shown in Table 1 below, and can be obtained by culturing a strain of Streptomyces kespitosus.

The three-dimensional structure is according to Journal of American Chemical Society, 105 , 7199 (1983).

In addition, 10th-position decarbamoylmitomycins are reported in Journal of Medicinal Chemistry, 14 , 109 (19
71).

Among these mitomycins, mitomycin C has a particularly strong antitumor activity and has been widely clinically used. However, it is highly toxic and myelotoxic, and various derivatives of mitomycins have been prepared so far for the purpose of enhancing antitumor activity and / or reducing side effects.

Among these derivatives, various mitomycins in which the 7-position amino group is substituted are known, but mitomycin derivatives substituted with a group in which an atom other than a carbon atom is arranged adjacent to the nitrogen atom of the amino group. As JP-A-60-1
Only those described in 69481 are known. That is, in the literature, the 7-position is methanesulfonylamino (Example 5), diethylthiophosphorylamino [(C ₂ H
₅ ) ₂ P (= S) NH-] (Example 14) and the like are disclosed. However, a mitomycin derivative in which the 7-position amino group is substituted with a group in which an oxygen atom or a nitrogen atom is arranged adjacent to the nitrogen atom is not known.

There is a constant need for new and useful mitomycin derivatives. As a result of research on a novel mitomycin derivative, it was found that a mitomycin derivative having a nitroso group at the 7-position or a substituted amino group having a nitrogen atom adjacent to the nitrogen atom and bonded by a double bond has excellent antitumor activity. It was These mitomycin derivatives are completely novel mitomycin derivatives in which the quinone ring of mitomycins is converted into a dihydroquinone ring.

Means for Solving Problems The mitomycin derivative of the present invention having excellent antitumor activity is represented by the following formula (I). : {In the formula, A is ON- or R ₄ N = N- [wherein, R ₄ is Or an unsubstituted or substituted heterocyclic group (in these formulas, R ₅ is a hydrogen atom, lower alkyl or lower cycloalkyl, R ₆ is a hydrogen atom, a halogen atom, hydroxy,
Lower alkoxy, amino or hitoro, R ₇ is lower alkyl or lower cycloalkyl, X is an oxygen atom, sulfur atom or imino, R ₈ is the same or different from R ₅ and is a hydrogen atom, lower alkyl or lower Is a cycloalkyl)], and R ₁ and R ₂ are the same or different and each is a hydrogen atom, lower alkyl, lower cycloalkyl, unsubstituted or substituted aralkyl, unsubstituted or substituted lower alkanoyl, unsubstituted or substituted arylcarbonyl, non-substituted. A substituted or substituted lower alkanesulfonyl, arylsulfonyl or aralkylsulfonyl, R ₃ is a hydrogen atom or carbamoyl, Y is a hydrogen atom or methyl, Z is a hydrogen atom, methyl or acetyl, Represents an α bond or a β bond}. The compound represented by the formula (I) is hereinafter referred to as compound (I) (the same applies to compounds having other formula numbers). In the definition of R ₅ , R ₇ , R ₈ , R ₁ and R ₂ of the formula (I), lower alkyl is linear or branched alkyl having 1 to 6 carbon atoms, for example, methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n
-Hexyl and the like, lower cycloalkyl includes cycloalkyl having 3 to 6 carbon atoms such as cyclopropyl, cyclopentyl, cyclohexyl and the like.

In the definition of R _{6 in} formula (I), lower alkoxy has 1 to 6 carbon atoms.
A straight chain or branched alkoxy of, for example, methoxy,
Includes ethoxy, isopropoxy, n-butoxy and the like.

In the definition of R ₄ , the heterocyclic group is a 5- or 6-membered monocyclic heterocyclic group having 1, 2 or 3 non-integrated double bonds having only 1 to 4 N, only one S or O. With 5 or 6
A monocyclic heterocyclic group having a 1 or 2 non-integrated double bond of a member ring, or a 1 or 2 non-integrated double of a 5- or 6-membered ring having one S or O and 1 or 2 N A monocyclic heterocyclic group having a bond and the like are included. Specific preferred heterocyclic groups include imidazolyl, imidazolinidyl, thiazolyl, thiazolinidyl, oxazolyl, oxazolidinyl, triazolyl, pyrimidyl and the like. The substituent of the heterocycle is lower alkyl (lower alkyl as used herein means
R ₅ , R ₇ , R ₈ , R ₁ and R ₂ have the same meanings).

In the definition of R ₁ and R ₂ , unsubstituted or substituted aralkyl includes benzine, phenethyl, diphenylmethyl, trityl, substituted benzyl and the like. Here, the substituent of the substituted benzyl is the substituent of the benzene nucleus, and is 1 or 2 the same or different lower alkoxy (lower alkoxy as used herein means R ₆
, Halogen atom (chlorine atom, fluorine atom, etc.), nitro, cyano, lower alkyl (lower alkyl as used herein has the same meaning as in R ₅ , R ₇ , R ₈ , R ₁ and R ₂₎ Is included) and the like. Regarding the definition of R ₁ and R ₂ ,
The lower alkanoyl referred to as an unsubstituted or substituted lower alkanoyl is a linear or branched alkanoyl having 1 to 6 carbon atoms, for example, formyl, acetyl, propionyl, N.
-Butyryl, isobutyryl, n-pentanoyl, n-hexanoyl and the like are included, and the substituent includes a halogen atom such as a chlorine atom and a fluorine atom. Specific examples of the substituted lower alkanoyl include trichloroacetyl, trifluoroacetyl and the like. With respect to the definitions of R ₁ and R _2, the aryl as defined in the unsubstituted or substituted arylcarbonyl includes phenyl, naphthyl and the like, and the substituent is 1 or 2 the same or different lower alkoxy (wherein the lower alkoxy as defined in R ₆ is Synonymous with), halogen atom (chlorine atom, fluorine atom, etc.), nitro, cyano, lower alkyl (lower alkyl as used herein is as defined in R ₅ , R ₇ , R ₈ , R ₁ and R ₂ ) Etc. are included. Regarding the definitions of R ₁ and R _2, the lower alkanesulfonyl referred to as an unsubstituted or substituted lower alkanesulfonyl is a linear or branched alkanesulfonyl having 1 to 6 carbon atoms, and the substituent is a halogen atom, for example, a chlorine atom, Including a fluorine atom and the like. Specific examples of the unsubstituted or substituted lower alkanesulfonyl include methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, n-butanesulfonyl, n-hexanesulfonyl, trichloromethanesulfonyl, trifluoromethanesulfonyl and the like. Regarding the definition of R ₁ and R ₂ , arylsulfonyl includes benzenesulfonyl, p-trienesulfonyl and the like, and aralkylsulfonyl includes benzylsulfonyl and the like.

In the formula (I), the compound in which R ₁ = R ₂ = H [hereinafter referred to as the compound (I-1)] is represented by the formula (II) (Wherein R ₃ , Y, Z and Is the same as in formula (I)) and hydroxylamine or an acid addition salt thereof or formula (III) R ₄ NHNH ₂ (III) (wherein R ₄ is as defined in formula (I)). It can be obtained by reacting a compound represented by the same meaning) or an acid addition salt thereof in an inert solvent, if necessary, in the presence of a base.

As the acid addition salt of hydroxylamine or compound (III), there are used inorganic acid salts such as hydrochloride and hydrobromide, and organic acid salts such as acetate, oxalate and p-toluenesulfonate. . The amount of hydroxylamine, compound (III) or acid addition salt thereof used is 1 to 3 relative to compound (II).
Equivalent weight is appropriate. When the acid addition salt of hydroxylamine or the acid addition salt of compound (III) is used, the amount of the base required to release hydroxylamine or compound (III) is required. Such bases include sodium carbonate, potassium carbonate, sodium hydrogen carbonate, sodium hydroxide,
Inorganic bases such as potassium hydroxide and lithium hydroxide, pyridine, 4-dimethylaminopyridine, triethylamine,
Tertiary amines such as N, N-dimethylaniline, alkali metal alkoxides such as sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide and the like are used.

In the case of reacting the compound (II) with the free compound (III), the combined use of a base may promote the reaction in a preferable direction.

As the inert solvent, lower alkanol such as methanol, ethanol and isopropanol, acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran, water and the like can be used alone or in combination.

The reaction is preferably carried out at 0 ° C. to room temperature, and usually completed in a few minutes to a few hours.

After completion of the reaction, the compound (I-1) can be obtained by a commonly used treatment operation such as extraction, chromatography, separation such as recrystallization, and a purification method.

It is considered that the compound (I-1) can have not only the structure of the formula (I) but also the structure of the following formula (IV) by tautomerism: [Wherein B is hydroxy or R ₄ NH (wherein R ₄ has the same meaning as in formula (I)), and R ₃ , Y, Z and Is as defined in formula (I)].

For example, the ¹ H- and ¹³ C-NMR values in d ₆ -dimethylsulfoxide of the compound obtained in Example 1 show that the compound has a structure corresponding to the formula (I). However, it is generally considered that the biological ratio of the tautomer differs depending on the type of compound, the measurement conditions of NMR and the like. Therefore, the present invention relates to the mitomycin derivative represented by the formula (I), and also includes the compound (IV) which is a tautomer thereof when R ₁ = R ₂ = H.

In formula (I), R ₁ and R ₂ are the same or different and each is a hydrogen atom,
A compound which is a lower alkyl, a lower cycloalkyl, or an unsubstituted or substituted aralkyl (except when both are hydrogen atoms) [hereinafter referred to as a compound (I-2)] is a compound represented by the formula (I-1) and a compound represented by the formula R _9- Hal, (R ₉ ) ₂ SO ₄ , C
An alkylating agent represented by H ₂ N ₂ or (R ₉ ) ₃ OBF ₄ (in each formula, R ₉ is lower alkyl, lower cycloalkyl, or unsubstituted or substituted aralkyl) in an inert solvent, It can be obtained by reacting in the presence of a base.

The alkylating agent is used in an amount of 0.8 to 1.2 with respect to compound (I-1).
When used in an equivalent amount, a compound in which only R ₂ is alkylated can be obtained, and when used in an amount of 2 to 4 equivalents, a compound in which both R ₁ and R ₂ are alkylated can be obtained. Examples of the base include sodium hydride, lithium hydride, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, inorganic bases such as silver oxide sodium methoxide, sodium ethoxide,
Organic bases such as potassium methoxide, potassium ethoxide, potassium tert-butoxide, n-butyl lithium, sec-butyl lithium, phenyl lithium and lithium diisopropylamide are used. The base is usually used in the same amount as the alkylating agent.

As the inert solvent, lower alkanol such as methanol, ethanol, isopropanol and tert-butanol; acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran and the like can be used alone or in combination.

In formula (I), R ₁ and R ₂ are the same or different and each is a hydrogen atom,
Unsubstituted or substituted lower alkanoyl, or unsubstituted or substituted arylcarbonyl (except when both are hydrogen atoms), and Z is not a hydrogen atom [hereinafter referred to as compound (I-3)], Z is hydrogen not atomic compound (wherein, R ₁₀ is unsubstituted or substituted lower alkanoyl or unsubstituted or a substituted arylcarbonyl) (I-1) and the formula R _{10 -OH} reactive derivative of the carboxylic acid represented by (hereinafter , An acylating agent) in the presence of a base in an inert solvent.

The reactive derivative referred to as an acylating agent includes acid halides such as acid chlorides and acid bromides, and acid anhydrides. When 0.8 to 1.2 equivalents of the acylating agent are used with respect to the compound (I-1), a compound in which only R ₂ is acylated is obtained,
When used in 2 to 4 equivalents, R ₁ and R ₂ coacylated compounds are obtained. Examples of the base include inorganic bases such as sodium hydride, lithium hydride, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, and silver oxide; sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium tert. -Butoxide, n
Organic bases such as -butyllithium, sec-butyllithium, phenyllithium, lithium diisopropylamide, triethylamine, pyridine, N, N-dimethylaniline and 4-dimethylaminopyridine are used. The base is usually used in the same amount as the acylating agent.

As the inert solvent, acetonitrile, dimethylformamide, dimethylsulfoxide, tetrahydrofuran or the like is used. When an acid anhydride is used as the acylating agent, a solvent may be added to a base such as triethylamine, pyridine and N, N-dimethylaniline.

In formula (I), R ₁ and R ₂ are the same or different and each is a hydrogen atom,
Unsubstituted or substituted lower alkanoyl, or unsubstituted or substituted arylcarbonyl (except when they are both hydrogen atoms) and Z is a hydrogen atom [hereinafter referred to as compound (I-3 ′)] is the above-mentioned compound. The compound (I-3) in which Z is acetyl obtained as described above is obtained by reacting with an aqueous solution of hydroxylamine in a lower alkanol. The amount of hydroxylamine used is appropriately 1 to 2 equivalents of the compound (I-3) in which Z is acetyl. Lower alkanols include methanol, ethanol, isopropanol and the like.

In formula (I), R ₁ and R ₂ are the same or different and each is a hydrogen atom,
A compound which is an unsubstituted or substituted lower alkanesulfonyl, arylsulfonyl, or aralkylsulfonyl (except when both are hydrogen atoms) [hereinafter referred to as compound (I-4)] is obtained by converting the compound (I-1) into the compound represented by the formula R ₁₁ -OH
(In the formula, R ₁₁ is unsubstituted or substituted lower alkanesulfonyl, arylsulfonyl or aralkylsulfonyl), and a reactive derivative of sulfonic acid (hereinafter,
And a sulfonylating agent) in the presence of a base in an inert solvent.

The reactive derivative referred to as a sulfonylating agent includes acid halides such as acid chloride and acid bromide, and acid anhydrides.
The sulfonylating agent is used in an amount of 0.8 to 1.
When 2 equivalents are used, a compound in which only R ₂ is sulfonylated is obtained, and when 2 to 4 equivalents are used, a compound in which both R ₁ and R ₂ are sulfonylated is obtained. The base, the amount of the base used, and the inert solvent may be the same as in the acylation.
When an acid anhydride is used as the sulfonylating agent, a solvent may be added to a base such as triethylamine, pyridine, N, N-dimethylaniline.

The above alkylation, acylation, partial deacylation, and sulfonylation reactions are suitably carried out at 0 ° C. to room temperature, and are usually completed in several hours to several tens of hours.

After completion of each reaction, the compound (I-2), (I-3) or (I-4) can be obtained by a commonly used treatment operation, for example, separation such as extraction, chromatography, recrystallization, etc., or a purification method. .

Compound (I) shows excellent antitumor activity. Hereinafter, the pharmacological action of the representative compound (I) will be shown in Experimental Examples.

Experimental Example 1 Table 2 shows the effects of representative compound (I) on Hela S ₃ cultured cells. In the table, IC ₅₀ is the concentration of the test compound that gives a cell number of 50% with respect to the cell number without drug administration.

The test method is as follows.

Cultured Hela S ₃ cells were 3 × 10 5 in MEM medium containing 10% (V / V) fetal bovine serum and 292 mg / ml glutamine.
^The cell suspension was prepared at ⁴ cells / ml. Dispense 1 ml each into a 24-well multi-plate and use a carbon dioxide incubator (5% carbon dioxide,
Incubated at 37 ° C. in 95% air). PBS after 24 hours
Alternatively, drugs dissolved or suspended in ethanol or dimethyl sulfoxide were added at various concentrations. After the addition, the cells were cultured for 72 hours in a 37 ° C. carbon dioxide incubator (5% carbon dioxide, 95% air). The culture solution in each well was removed by suction with an aspirator, 1 ml of PBS was added, the cell surface was gently washed, and then removed by suction. 0.05% trypsin and 0.02% E
1 ml of DTA-containing PBS was dispensed into each well, and after pipetting, the number of cells in each cell suspension was counted with a microcell counter (Toa Medical Electronics). Concentration showing 50% of the number of cells without drug administration (IC
₅₀ ) was calculated from the concentration-dependent curve.

Experimental example 2. Table 3 shows the effects of representative compound (I) on Sarcoma 180 solid tumor. CI in the table means the chemotherapy coefficient It is represented by. LD ₅₀ here is the acute toxicity value and ED ₅₀
Indicates the dose that reduces the volume of Sarcoma 180 solid tumor to 50% of the tumor volume of the non-administration control group.

In the table Is the dose and ED ₅₀ that give a peripheral leukocyte count of 4000.
And shows the effect on the peripheral white blood cell count.

The values of LD ₅₀ , ED ₅₀ and WBC ₄₀₀₀ were obtained by the methods described below.

(1) LD ₅₀ ddy mice were intraperitoneally administered with a drug once, and the mice were observed for life and death 14 days after the administration of 5 mice per group. From the mortality rate of each administration group, LD _{50 was} measured according to the Behrens-Kelber method. Was calculated.

(2) ED ₅₀ 5 × 10 ⁶ sarcoma 180 cells were intraperitoneally transplanted into a ddy mouse, cells were collected from ascites on day 7, washed once with sterile physiological saline, and then with sterile physiological saline. A cell suspension of 5 × 10 ⁷ cells / ml was prepared. This 0.1 ml weight 20
± 2 g of ddy male mouse was subcutaneously transplanted into the right axilla.
The drug was dissolved in physiological saline or physiological saline containing Tween 80, and 0.1 to 0.2 ml was administered from the tail vein of 5 mice per group 24 hours after tumor implantation. Major axis of the tumor 7 days after implantation (a) and minor axis (b) was measured to determine the value of a × b ^2/2 corresponding to the tumor volume. The ratio of the volume (T) of the drug-administered group to the volume (C) of the non-administered control group (T)
/ C) indicates the antitumor effect. T / C at each dose was plotted in a graph in which the vertical axis represents T / C on a normal scale and the horizontal axis represents dose on a logarithmic scale.
The relationship of / C was obtained as a straight line by the method of least squares. The dose (ED ₅₀ ) at which T / C was 0.5 was calculated from the obtained linear regression equation.

(3) WBC ₄₀₀₀ 5 × 10 ⁶ sarcoma 180 cells were subcutaneously transplanted into the right axillary region of 5 ddy male mice each weighing 20 ± 2 g per group, and the drug was intraperitoneally administered 24 hours later. 0.02 ml of blood from the orbital venous plexus of tumor-bearing mice on the 4th day after drug administration
It was collected and dispersed in 9.98 ml of Cell Kit Seven solution. After adding 1 drop of saponin solution to lyse the red blood cells, the white blood cell count was measured with a microcell counter. The vertical axis represents the number of peripheral leukocytes on a normal scale, and the horizontal axis represents the dose on a logarithmic scale. The number of leukocytes at each dose was plotted to find the relationship between the dose and peripheral leukocyte count.
0 / mm ³ (almost half the number of peripheral leukocytes in normal mice)
The dose (WBC ₄₀₀₀ ) that gave

As mentioned above, the compound (I) generally has excellent antitumor activity. The CI value of some of the compounds (I) was higher than that of mitomycin C, which means that the dose range was wider than that of mitomycin C. Also, some compounds (I) have WBC ₄₀₀₀ / ED ₅₀ values higher than that of mitomycin C, which means that at a dose giving an equivalent ED ₅₀ , myelotoxicity is less than that of mitomycin C. .

Therefore, the compound (I) can be used as an antitumor agent containing the same, particularly as an antitumor agent containing an effective amount of the compound (I) and a pharmaceutical adjuvant. Here, the medicinal auxiliaries include commonly used diluents, excipients, disintegrants, binders, lubricants, bases and the like.

Compound (I) can be used in various dosage forms.
When used as an injection, a diluent commonly used in this field as a diluent, for example, after dissolving Compound (I) in ethanol (using a surfactant and a solubilizer together if necessary), is ethanol removed by suction? Or without, distilled water for injection,
Physiological saline; manufactured by mixing with an injectable distilled aqueous solution of glucose, fructose, mannitol, or the like. Alternatively, an injection solution obtained by freeze-drying an ethanol solution or a powder injection solution obtained by mixing compound (I) with sodium chloride may be used, and in these cases, they are dissolved before use. These injections are provided for intravenous administration, but intramuscular administration, intraarterial administration, intraperitoneal administration, intrathoracic administration and the like are also possible. The preparation for oral administration is produced by mixing and molding the compound (I) and a suitable excipient, a disintegrating agent, a binder, a lubricant and the like by a conventional method to give tablets, granules and powders. The suppository formulation is prepared by mixing and molding the compound (I) and a commonly used carrier according to a conventional method.

The dose varies depending on the administration method, type of compound (I), age, symptoms, etc., but generally for mammals including humans,
2-150 mg / 60 kg is suitable as the compound (I) per day.

Examples Examples and reference examples of the present invention will be shown below. MASS
The spectrum is measured by FAB (Fast Atom Bombardment) method.

The nomenclature of the target compound in each example is different from that in Table 1. For example, the compound of Example 1 is represented by the following structure and name.

[1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-5-methyl-6-nitroso-
Azirino [2 ', 3': 3,4] pyrrolo [1,2-a]
Indole Example 1. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-5-methyl-6-nitroso-
Azirino [2 ', 3': 3,4] pyrrolo [1,2-a]
Manufacture of indole 2.38 g of hydroxylamine hydrochloride dissolved in 10 ml of water
Then add 2.18 g of anhydrous sodium carbonate little by little while stirring.
Add one. This solution was mixed with 6.00 g of mitomycin A.
Was added to 400 ml of methanol and added at room temperature for 2 hours.
Stir for a while. The solvent was distilled off under reduced pressure, and the residue was silica gel.
Chromatography chloroform-methanol
Separation and purification using (9: 1; V / V) as developing solvent, and labeling
5.12 g of a compound is obtained (yield 86.5%). ¹ H-NMR (d ₆ -DMSO) δ; 2.06 (3H, s), 2.76 (1H, dd), 2.89 (1
H, d), 3.16 (3H, s), 3.45 (1H, dd), 3.53 (1H, dd), 4.09 (1H, d)
d), 4.17 (1H, d), 4.63 (1H, dd), 6.55 (2H, br), 10.17 (1H, s),
19.17 (1H, s) ¹³ C-NMR (d ₆ -DMSO) δ; 9.5, 31.2, 35.5, 41.6, 49.3, 49.
6, 60.0, 105.9, 107.7, 120.9, 139.0, 148.2, 156.5, 160.1, 1
69.0 IR (KBr) cm ^-1 3400, 2940, 1705, 1590, 1480, 1330, 1210, 106
0 MASS 351 (C ₁₅ H ₁₈ N _Four O ₆ Molecular weight 350.34) Example 2. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,5-dimethyl-1,1a, 2,8,
8a, 8b-hexahydro-8a-methoxy-b-nit
Loso-Azirino [2 ', 3': 3,4] pyrrolo [1,2]
-A] Production of indole By the same method as in Example 1, hydroxylamine hydrochloric acid was added.
0.038g salt, 0.033g anhydrous sodium carbonate and
Itomycin F 0.100 g to the title compound 0.098
g is obtained (yield 87.0%). ¹ H-NMR (d ₆ -DMSO) δ; 2.05 (3H, s), 2.26 (3H, s), 2.36 (1
H, dd), 2.40 (1H, d), 3.22 (3H, s) 3.58 (1H, dd), 3.67 (1H, d
d), 4.10 (1H, d), 4.15 (1H, dd), 4.68 (1H, dd), 6.40 (2H, br),
9.98 (1H, s), 19.05 (1H, s) IR (KBr) cm ^-1 3400, 2950, 1705, 1590, 1480, 1340, 1220, 106
0 MASS 365 (C ₁₆ H ₂₀ N _Four O ₆ Molecular weight 364.36) Example 3. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -1,5-
Dimethyl-1,1a, 2,8,8a, 8b-hexahydr
Ro-6-nitroso-4,7,8a-trihydroxy-a
Dilino [2 ', 3': 3,4] pyrrolo [1,2-a] ii
Preparation of Ndole Hydroxylamine hydrochloride was prepared in the same manner as in Example 1.
0.49 g salt, 0.45 g anhydrous sodium carbonate and
Itomycin B 1.20 g to the title compound 0.88 g
Is obtained (yield 73.2%). ¹ H-NMR (d ₆ -DMSO) δ; 2.04 (3H, s), 2.24 (3H, s), 2.35 (1
H, dd), 2.49 (1H, d), 3.63 (1H, dd) 3.72 (1H, dd), 4.08 (1H,
d), 4.15 (1H, dd), 4.74 (1H, dd), 6.38 (2H, br), 6.62 (1H, b
r), 9.52 (1H, s), 19.34 (1H, s) IR (KBr) cm ^-1 3400, 2950, 1705, 1595, 1480, 1345, 1220, 105
5 MASS 351 (C ₁₅ H ₁₈ N _Four O ₆ Molecular weight 350.34) Example 4. [1aS- (1aα, 8β, 8aα, 8bα)]-4,
7-dihydroxy-1,1a, 2,8,8a, 8b-f
Xahydro-8-hydroxymethyl-8a-methoxy-
5-methyl-6-nitroso-azirino [2 ', 3':
Production of 3,4] pyrrolo [1,2-a] indole By the same method as in Example 1, decarbamoyl mitoma was prepared.
Isin A (Journal of Medicinal Chemist
Lee, vol. 14, p. 109, 1971) 0.13
g, hydroxylamine hydrochloride 0.059 g anhydrous sodium carbonate
0.14 g of the title compound is obtained from 0.054 g of thorium.
(Yield 85.3%). ¹ H-NMR (CDCl _Three / CD _Three OD) δ; 2.19 (3H, s), 2.9
0 (1H, dd), 2.95 (1H, d), 3.23 (3H, s), 3.44 (1H, dd), 3.63 (1
H, dd), 3.98 (1H, dd), 4.04 (1H, dd), 4.35 (1H, d), 18.76 (1H,
s) IR (KBr) cm ^-1 3450, 2950, 1595, 1530, 1485, 1380, 1330, 121
0, 1030 MASS 308 (C ₁₄ H ₁₇ N ₃ O _Five Molecular weight 307.31) Example 5. [1aS- (1aα, 8β, 8aα, 8bα)]-4,
7-dihydroxy-1,5-dimethyl-1,1a, 2
8,8a, 8b-Hexahydro-8-hydroxymethyl
-8a-methoxy-6-nitroso-azirino [2 ',
Production of 3 ′: 3,4] pyrrolo [1,2-a] indole By the same method as in Example 1, decarbamoyl mitoma was prepared.
Isin F (Chemical Abstracts, Vol. 74, 999
20n, 1971) 0.30g, hydroxylamine
Hydrochloride 0.14 g and anhydrous sodium carbonate 0.10 g
From the above, 0.21 g of the title compound is obtained (yield 69.5%). ¹ H-NMR (CDCl _Three / CD _Three OD) δ; 2.10 (3H, s), 2.3
1 (3H, s), 2.37 (1H, dd), 2.40 (1H, d), 3.21 (3H, s), 3.39 (1H,
dd), 3.58 (1H, dd), 3.94 (1H, dd), 4.01 (1H, dd), 4.35 (1H,
d), 18.77 (1H, s) IR (KBr) cm ^-1 3450, 2950, 1590, 1530, 1480, 1370, 1210, 103
0 MASS 322 (C ₁₅ H ₁₉ N ₃ O _Five Molecular weight 321.33) Example 6. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,8a
-Dimethoxy-1,1a, 2,8,8a, 8b-hexa
Hydro-7-hydroxy-5-methyl-6-nitroso-
Azirino [2 ', 3': 3,4] pyrrolo [1,2-a]
Production of indole 3.00 g of the compound obtained in Example 1 was added to dimethylform
Dissolve in 50 ml of amide and 50 ml of tert-butanol,
-Add 0.86 g of butoxypotassium and stir for 20 minutes at room temperature.
Stir. 0.69 ml of methyl iodide was added, and at room temperature
Stir for 1.5 hours. The oil obtained by distilling off the solvent under reduced pressure
Chloroform and water were added to the product to extract chloroform.
The layer was dried over anhydrous sodium sulfate and then subjected to chloroform under reduced pressure.
The solvent was distilled off and the residue was chromatographed on silica gel.
Chloroform-methanol (15: 1; v / v) as the developing solvent
The product is separated and purified to give 1.32 g of the title compound (yield 4
2.3%). ¹ H-NMR (CDCl _Three ) Δ: 2.30 (3H, s), 2.87 (1H, dd), 2.9
8 (1H, d), 3.24 (3H, s), 3.57 (1H, dd), 3.72 (3H, s), 3.74 (1H, d
dd), 4.24 (1H, d), 4.51 (1H, dd), 4.84 (1H, dd), 19.01 (1H, s) IR (KBr) cm ^-1 3400, 2930, 1715, 1605, 1480, 1335, 1220, 106
5 MASS 365 (C ₁₆ H ₂₀ N _Four O ₆ Molecular weight 364.37) Example 7. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,8a
-Dimethoxy-1,5-dimethyl 1,1a, 2,8,8
a, 8b-Hexahydro-7-hydroxy-6-nitro
So-azirino [2 ', 3': 3,4] pyrrolo [1,2-
a] Preparation of indole 0.30 g of the compound obtained in Example 2 and tert-but
Xypotassium 0.10 g and methyl iodide 0.07 ml
Was reacted in the same manner as in Example 6 to obtain 0.12 g of the title compound.
Obtained (yield 38.7%). ¹ H-NMR (CDCl _Three ) Δ: 2.28 (3H, s), 2.29 (3H, s), 2.36
(1H, dd), 2.38 (1H, d), 3.22 (3H, s), 3.51 (1H, dd), 3.66 (1H,
dd), 3.72 (3H, s), 4.22 (1H, d), 4.32 (1H, dd), 4.78 (2H, br),
4.84 (1H, dd), 19.03 (1H, s) IR (KBr) cm ^-1 3400, 2950, 1720, 1605, 1480, 1340, 1215, 115
0, 1055 MASS 379 (C ₁₇ H _{twenty two} N _Four O ₆ Molecular weight 378.40) Example 8. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -1,1
a, 2,8,8a, 8b-hexahydro-5-methyl-
6-nitroso-4,7,8a-trimethoxy-azirino
[2 ', 3': 3,4] pyrrolo [1,2-a] indole
Preparation of 0.30 g of the compound obtained in Example 1 in 15 ml of DMF
Dissolve, sodium hydride (60% oil) 0.069
g and stirred at room temperature for 30 minutes, methyl iodide 0.12
Add ml and stir at room temperature for 1 hour. Solvent under reduced pressure
After distilling off, chloroform and water were added to the obtained oily matter, and the oily matter was extracted.
The extracted chloroform layer was dried over anhydrous sodium sulfate,
Chloroform was distilled off under reduced pressure, and the residue was washed with silica gel.
Chromatography-methanol (15:
1; v / v) is used as a developing solvent to separate and purify the title compound 0.10.
g is obtained (yield 30.8%). ¹ H-NMR (d ₆ -DMSO) δ: 2.14 (3H, s), 2.44 (1H, d
d), 2.48 (1H, d), 3.38 (3H, s), 3.49 (1H, dd), 3.66 (3H, s), 3.
88 (1H, dd), 3.95 (1H, d), 4.06 (3H, s), 4.49 (1H, dd), 4.55 (1
H, dd), 6.55 (2H, br) IR (KBr) cm ^-1 3450, 2940, 1705, 1620, 1600, 1540, 1450, 133
0, 1220, 1050 MASS 379 (C ₁₇ H _{twenty two} N _Four O ₆ Molecular weight 378.40) Example 9. [1aS- (1aα, 8β, 8aα, 8bα)]-4,
8a-dimethoxy-1,1a, 2,8,8a, 8b-f
Xahydro-7-hydroxy-8-hydroxymethyl-
5-methyl-6-nitroso-azirino [2 ', 3':
Preparation of 3,4] pyrrolo [1,2-a] indole Journal from 0.22 g of the compound obtained in Example 6
Of Medicinal Chemistry, Volume 14, 109 pages
, Decarbamoylation by the method described in 1971
Then, 0.14 g of the title compound is obtained (yield 72.7%). ¹ H-NMR (CDCl _Three ) Δ: 2.28 (3H, s), 2.91 (1H, dd), 2.9
8 (1H, d), 3.30 (3H, s), 3.48 (1H, dd), 3.58 (1H, dd), 3.78 (3
H, s), 4.03 (1H, dd), 4.08 (1H, dd), 4.24 (1H, d), 18.90 (1H,
s) IR (KBr) cm ^-1 3400, 2930, 1600, 1460, 1380, 1205, 1035 MASS 322 (C ₁₅ H ₁₉ N ₃ O _Five Molecular weight 321.33) Example 10. [1aS- (1aα, 8α, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -7,8a
-Dihydroxy-1,5-dimethyl-1,1a, 2
8,8a, 8b-Hexahydro-4-methoxy-6-ni
Toroso-azirino [2 ', 3': 3,4] pyrrolo [1,
2-a] Preparation of indole 0.90 g of the compound obtained in Example 3, tert-butoxy
Add 0.30 g of potassium and 0.22 ml of methyl iodide.
The reaction is carried out in the same manner as in Example 6 to obtain 0.28 g of the title compound.
(Yield 29.9%). ¹ H-NMR (CDCl _Three / CD _Three OD) δ: 2.24 (3H, s), 2.3
2 (3H, s), 2.38 (1H, dd), 2.64 (1H, d), 3.60 (1H, dd), 3.70 (3
H, s), 3.76 (1H, dd), 4.15 (1H, d), 4.59 (1H, dd), 4.82 (1H, d)
d), 19.15 (1H, s) IR (KBr) cm ^-1 3420, 2950, 1700, 1605, 1520, 1480, 1350, 106
0 MASS 365 (C ₁₆ H ₂₀ N _Four O ₆ Molecular weight 364.37) Example 11. [1aS- (1aα, 8β, 8aα, 8bα)]-1-
Acetyl-4-acetyloxy-8-{[(aminocal
Bonyl) oxy] methyl} -1,1a, 2,8,8a,
8b-hexahydro-7-hydroxy-8a-methoxy
-5-methyl-6-nitroso-azirino [2 ', 3':
Production of 3,4] pyrrolo [1,2-a] indole N'-acetylmitomycin A (Japanese Patent Publication No. 40-181)
77) as a raw material and obtained in the same manner as in Example 1 [1
aS- (1aα, 8β, 8aα, 8bα)]-1-ace
Cyl-8-{[(aminocarbonyl) oxy] methyl}
-4,7-Dihydroxy-1,1a, 2,8,8a, 8
b-hexahydro-8a-methoxy-5-methyl-6-
Nitroso-azirino [2 ', 3': 3,4] pyrrolo
1.03 g of [1,2-a] indole was added to pyridine 110
Dissolve in 0.3 ml, add 0.3 ml of acetic anhydride and stir at room temperature for 10 hours
Stir. The oily substance obtained by distilling off the solvent under reduced pressure is treated with silica gel.
Chromatography-chloroform-methanol
Separated and purified using (15: 1; v / v) as a developing solvent,
1.00 g of the compound is obtained (yield 87.6%). ¹ H-NMR (CDCl _Three ) Δ: 2.14 (3H, s), 2.32 (3H, s), 2.40
(3H, s), 3.19 (3H, s), 3.30 (1H, dd), 3.48 (1H, dd), 3.53 (1H,
d), 3.77 (1H, dd), 4.01 (1H, dd), 4.09 (1H, d), 4.99 (1H, dd),
15.80 (1H, s) IR (KBr) cm ^-1 3400, 2950, 1710, 1620, 1480, 1340, 1210, 119
0, 1070 MASS 435 (C ₁₉ H _{twenty two} N _Four O ₈ Molecular weight 434.40) Example 12. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -1,1
a, 2,8,8a, 8b-hexahydro-7-hydroxy
Ci-4-isoprolyloxy-8a-methoxy-5-methyl
Lu-6-nitroso-azirino [2 ', 3': 3,4] pi
Production of Lolo [1,2-a] indole 0.50 g of the compound obtained in Example 1 was added to dimethylform
Dissolve in 20 ml of amide, sodium hydride (60% oil
Substance) 0.052 g, stirred at room temperature for 15 minutes, and then iodinated
Add 0.18 ml of isopropyl and 1.5 hours at room temperature
Stir for a while. The solvent was distilled off under reduced pressure, and the resulting oily substance was mixed with chlorine.
Add chloroform and water and extract the chloroform layer.
After drying over sodium, the chloroform was distilled off under reduced pressure,
Chromatograph the residue on silica gel chromatography.
Rum-methanol (30: 1; v / v) as developing solvent
Separation and purification give 0.16 g of the title compound (yield 28.
5%). ¹ H-NMR (CDCl _Three ) Δ: 1.32 (6H, d), 2.26 (3H, s), 2.85
(1H, dd), 2.98 (1H, d), 3.22 (3H, s), 3.50 (1H, dd), 3.69 (1H,
dd), 4.20 (1H, q), 4.28 (1H, d), 4.43 (1H, dd), 4.87 (1H, dd),
5.10 (2H, br), 19.06 (1H, s) IR (KBr) cm ^-1 3450, 2950, 1710, 1600, 1480, 1335, 1210, 106
0 MASS 393 (C ₁₈ H _{twenty four} N _Four O ₆ Molecular weight 392.41) Example 13. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -1,1
a, 2,8,8a, 8b-hexahydro-7-hydroxy
Ci-4-isobutyloxy-8a-methoxy-5-methyl
Lu-6-nitroso-azirino [2 ', 3': 3,4] pi
Production of Lolo [1,2-a] indole 0.50 g of the compound obtained in Example 1 was treated with hydrogenated sodium chloride.
0.057 g of um (60% oil) and isobutane iodide
Chill 0.25 ml was reacted as in Example 12 and the title compound
0.11 g of the product is obtained (yield 18.1%). ¹ H-NMR (CDCl _Three ) Δ: 1.06 (3H, d), 1.07 (3H, d), 2.07
(1H, m), 2.27 (3H, s), 2.86 (1H, dd), 2.99 (1H, d), 3.24 (3H,
s), 3.45 (1H, d), 3.51 (1H, d), 3.51 (1H, dd), 3.65 (1H, d), 3.
69 (1H, dd), 4.24 (1H, d), 4.46 (1H, dd), 4.84 (1H, dd), 5.05
(2H, br), 19.05 (1H, s) IR (KBr) cm ^-1 3450, 2950, 1710, 1600, 1480, 1340, 1220, 106
0 MASS 407 (C ₁₉ H ₂₆ N _Four O ₆ Molecular weight 406.44) Example 14. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4-ben
Ziroxy-1,1a, 2,8,8a, 8b-hexahi
Doro-7-hydroxy-8a-methoxy-5-methyl-
6-nitroso-azirino [2 ', 3': 3,4] pyrrolo
Production of [1,2-a] indole 0.42 g of the compound obtained in Example 1 was added to dimethylform
It was dissolved in 9 ml of amide and 9 ml of tert-butanol and te under ice cooling.
Add 0.12 g of rt-butoxypotassium and stir for 30 minutes
It After adding 0.15 ml of benzyl bromide under ice cooling, 1 at room temperature
Stir for hours. The solvent was distilled off under reduced pressure to give an oily product.
Add chloroform and water to the extracted chloroform layer.
After drying over sodium acidate, chloroform was distilled off under reduced pressure.
The residue is chromatographed by silica gel chromatography.
Rhoform-methanol (20: 1; v / v) as developing solvent
The product is separated and purified to give 0.09 g of the title compound (yield 1
7.5%). ¹ H-NMR (CDCl _Three ) Δ: 2.27 (3H, s), 2.80 (1H, dd), 2.9
5 (1H, d), 3.17 (3H, s), 3.37 (1H, dd), 3.71 (1H, dd), 4.23 (1
H, d), 4.52 (1H, dd), 4.70 (2H, br), 4.85 (2H, s), 4.86 (1H, d)
d), 7.39 (5H, s), 18.99 (1H, s) IR (KBr) cm ^-1 3450, 2950, 1700, 1630, 1590, 1480, 1320, 120
5, 1060 MASS 441 (C _{twenty two} H _{twenty four} N _Four O ₆ Molecular weight 440.46) Example 15. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -1,1
a, 2,8,8a, 8b-hexahydro-7-hydroxy
Ci-8a-methoxy-5-methyl-6-nitroso-4-
[(P-toluenesulfonyl) oxy] azirino
[2 ', 3': 3,4] pyrrolo [1,2-a] indole
Preparation of 0.33 g of the compound obtained in Example 1
Dissolve in 40 ml of orchid and add 0.1% triethylamine under ice cooling.
2 ml and 0.15 g of p-toluenesulfonic acid chloride
In addition, the mixture is stirred at the same temperature for 30 minutes. The reaction solution is saturated saline
The tetrahydrofuran layer with anhydrous sodium sulfate.
After drying, concentrate under reduced pressure and perform silica gel chromatography.
-Chloroform-methanol (30: 1; v / v)
0.11 g of the title compound as a developing solvent
(Yield 22.8%). ¹ H-NMR (CDCl _Three ) Δ: 2.17 (3H, s), 2.49 (3H, s), 2.88
(1H, dd), 2.98 (1H, d), 3.31 (3H, s), 3.66 (1H, d), 3.72 (1H, d
d), 4.12 (1H, d), 4.48 (1H, dd), 4.70 (2H, br), 4.83 (1H, dd),
7.40 (2H, d), 7.82 (2H, d), 18.43 (1H, s) IR (KBr) cm ^-1 3460, 2930, 1715, 1620, 1460, 1390, 1335, 119
5, 1180 MASS 505 (C _{twenty two} H _{twenty three} N _Four O ₈ S molecular weight 504.46) Example 16. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
[(Di-p-toluenesulfonyl) oxy] -1,1
a, 2,8,8a, 8b-hexahydro-8a-methoki
Ci-5-methyl-6-nitroso-azirino [2 ',
Preparation of 3 ': 3,4] pyrrolo [1,2-a] indole 0.27 g of the compound obtained in Example 1
Dissolve in 40 ml of orchid and add 0.3 of triethylamine under ice cooling.
3 ml and 0.38 g of p-toluenesulfonic acid chloride
The mixture is stirred at the same temperature for 2.5 hours. The reaction solution was used in Example 15.
Is treated in the same manner as above to obtain 0.11 g of the title compound (yield 2
1.5%). ¹ H-NMR (CDCl _Three ) Δ: 2.18 (3H, s), 2.44 (3H, s), 2.51
(3H, s), 2.96 (1H, dd), 3.03 (1H, d), 3.23 (3H, s), 3.50 (1H, d)
d), 3.60 (1H, dd), 3.96 (1H, d), 4.43 (1H, dd), 4.69 (1H, dd),
4.93 (2H, br), 7.31 (2H, d), 7.40 (2H, d), 7.82 (2H, d), 7.84
(2H, d) IR (KBr) cm ^-1 3460, 2925, 1715, 1640, 1615, 1485, 1460, 139
0, 1340, 1195, 1180 MASS 659 (C ₂₉ H ₃₀ N _Four O _Ten S ₂ Molecular weight 658.59) Example 17. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Benzoyl) azo] -4,7-dihydroxy-1,
1a, 2,8,8a, 8b-hexahydro-8a-meth
Xy-5-methyl-azirino [2 ', 3': 3,4] pi
Production of Lolo [1,2-a] indole Mitomycin A (1.00 g) was dissolved in methanol and stirred.
1.17 g of benzoylhydrazine in this solution with stirring
Further add 0.10 g of potassium carbonate and stir at room temperature for 3 hours.
Stir. The solvent was distilled off under reduced pressure and the residue was washed with silica gel.
Chromatography-chloroform-methanol
(6: 1; v / v) was used as a developing solvent for separation and purification.
1.04 g of the compound is obtained as a tan powder (yield 80.
1%). ¹ H-NMR (CDCl _Three / D ₆ -DMSO) δ: 2.19 (3H,
s), 2.83 (1H, m), 3.00 (1H, m), 3.24 (3H, s), 3.59 (1H, d), 3.6
3 (1H, dd), 4.32 (1H, d), 4.39 (1H, dd), 4.83 (1H, dd), 5.61 (2
H, br), 7.54 (3H, m), 8.00 (2H, dd), 8.96 (1H, br), 17.04 (1H,
s) ¹³ C-NMR (CDCl _Three / D ₆ -DMSO) delta: 10.3, 31.
7, 36.2, 42.5, 49.3, 49.8, 60.8, 105.6, 109.3, 120.8, 127.
6, 128.8, 132.6, 140.7, 141.6, 156.8, 156.9, 164.4, 173.5 IR (KBr) cm ^-1 3430, 3204, 1707, 1652, 1611, 1578, 1481, 145
1, 1381, 1328, 1240, 1154, 1068, 922 MASS 454 (C _{twenty two} H _{twenty three} N _Five O ₆ Molecular weight 453.46) Example 18. [1aS- (1aα, 8β, 8aα, 8bα)]-6-
[(Acetyl) azo] -8-{[(aminocarbonyl)
[Oxy] methyl} -4,7-dihydroxy-1,1a,
2,8,8a, 8b-hexahydro-8a-methoxy-
5-methyl-azirino [2 ', 3': 3,4] pyrrolo
Production of [1,2-a] indole By the same method as in Example 17, mitomycin A2
05 mg, acetohydrazide 130 mg and potassium carbonate 1
120 mg of the title compound is obtained from 00 mg (yield 52.2).
%). ¹ H-NMR (d ₆ -DMSO) δ: 2.01 (3H, s), 2.24 (3H, s),
2.75 (1H, m), 2.83 (1H, d), 3.14 (3H, s), 3.49 (1H, d), 4.07 (1
H, t), 4.07 (1H, d), 4.63 (1H, dd), 6.55 (2H, br), 15.37 (1H,
s) IR (KBr) cm ^-1 3450, 3182, 1699, 1618, 1552, 1484, 1378, 132
7, 1193, 1167, 1061, 993 MASS 392 (C ₁₇ H _{twenty one} N _Five O ₆ Molecular weight 391.38) Example 19. [1aS- (1aα, 8β, 8aα, 8bα)]-6-
[(4 "-aminobenzoyl) azo] -8-{[(ami
Nocarbonyl) oxy] methyl} -4,7-dihydroxy
Ci-1,1a, 2,8,8a, 8b-hexahydro-8
a-Methoxy-5-methyl-azirino [2 ', 3':
Production of 3,4] pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A30 was prepared.
2 mg, 4-aminobenzoylhydrazine and potassium carbonate
163 mg of the title compound (yield 40.
5%). ¹ H-NMR (d ₆ -DMSO) δ: 2.08 (3H, s), 2.76 (1H, d
d), 2.85 (1H, d), 3.15 (3H, s), 3.50 (1H, d), 4.11 (1H, d), 4.1
1 (1H, dd), 4.69 (1H, dd), 6.07 (2H, s), 6.53 (1H, dd), 6.58 (2
H, br), 6.66 (2H, d), 7.63 (2H, d), 16.93 (1H, s) IR (KBr) cm ^-1 3380, 1710, 1603, 1480, 1400, 1256, 1185, 106
8, 928 MASS 469 (C _{twenty two} H _{twenty four} N ₆ O ₆ Molecular weight 468.46) Example 20. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-6-[(4 "-hydroxybenzoyl) azo]
-8a-methoxy-5-methyl-azirino [2 ',
Production of 3 ′: 3,4] pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A30 was prepared.
1 mg, 4-hydroxybenzoylhydrazine 126 mg and
222 mg of the title compound is obtained from 107 mg of potassium carbonate.
(Yield 58.2%). ¹ H-NMR (d ₆ -DMSO) δ: 2.07 (3H, s), 2.76 (1H, d),
2.86 (1H, d), 3.15 (3H, s), 3.51 (1H, d), 4.09 (1H, dd), 4.12
(1H, d), 4.69 (1H, dd), 6.56 (2H, br), 6.95 (2H, d), 7.78 (2H, d
d), 17.01 (1H, s) IR (KBr) cm ^-1 3400, 1710, 1610, 1480, 1390, 1242, 1175, 107
8, 928 MASS 470 (C _{twenty two} H _{twenty three} N _Five O ₇ Molecular weight 469.45) Example 21. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Benzoyl) azo] -4,7-dihydroxy-1.
5-dimethyl-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-azirino [2 ', 3': 3,
4] Production of Pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin F20 was prepared.
0 mg, benzoylhydrazine 229 mg and potassium carbonate
230 mg of the title compound is obtained from 77 mg (yield 89.5).
%). ¹ H-NMR (CDCl _Three / D ₆ -DMSO) δ: 2.21 (3H,
s), 2.27 (1H, dd), 2.28 (3H, s), 2.40 (1H, d), 3.22 (3H, s), 3.
56 (1H, dd), 3.63 (1H, dd), 4.30 (1H, dd), 4.32 (1H, d), 4.84
(1H, dd), 5.19 (2H, br), 7.53 (3H, m), 8.02 (2H, dd), 8.63 (1
H, br), 17.01 (1H, s) IR (KBr) cm ^-1 3430, 3204, 1708, 1569, 1484, 1398, 1346, 123
7, 1156, 1067, 936 MASS 468 (C _{twenty three} H _{twenty five} N _Five O ₆ Molecular weight 467.49) Example 22. [1aS- (1aα, 8β, 8aα, 8bα)]-6-
[(Benzoyl) azo] -4,7-dihydroxy-1,
5-dimethyl-1,1a, 2,8,8a, 8b-hexa
Hydro-8-hydroxymethyl-8a-methoxy-azide
Rhino [2 ', 3': 3,4] pyrrolo [1,2-a] yne
Preparation of doll By the same method as in Example 17, decarbamoylmite was prepared.
Mycin F (CA, 74 , 99920n: 1971) 10
0 mg, benzoylhydrazine 129 mg and potassium carbonate
88 mg of the title compound is obtained from 43 mg (yield: 66.9).
%). ¹ H-NMR (CDCl _Three ) Δ: 2.17 (3H, s), 2.17 (1H, m), 2.18
(3H, s), 2.18 (1H, m), 3.18 (3H, s), 3.42 (1H, dd), 3.54 (1H, m
d), 4.01 (1H, m), 4.00 (1H, dd), 7.55 (3H, m), 7.99 (2H, m), 1
6.83 (1H, s) IR (KBr) cm ^-1 3400, 2876, 1674, 1483, 1395, 1349, 1236, 115
3, 1067, 943 MASS 425 (C _{twenty two} H _{twenty four} N _Four O _Five Molecular weight 424.46) Example 23. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-6 [(2 "-imidazolidino) azo] -8a-
Methoxy-5-methyl-azirino [2 ', 3': 3,
4] Production of Pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A1.
01 g, 3-hydrazino-2-imidazoline hydrobromic acid
The title compound was prepared from 0.48 g of salt and 0.25 g of potassium carbonate.
0.40 g of the product is obtained (yield 36.8%). ¹ H-NMR (d ₆ -DMSO) δ: 2.08 (3H, s), 2.78 (1H, m),
2.89 (1H, m), 3.16 (3H, s), 3.55 (1H, d), 3.80 (4H, s), 4.04 (1
H, dd), 4.15 (1H, d), 4.72 (1H, dd), 6.52 (2H, br), 16.51 (1H,
br) IR (KBr) cm ^-1 3400, 1710, 1640, 1578, 1488, 1410, 1338, 121
5, 1078, 1037, 1000, 910 MASS 418 (C ₁₈ H _{twenty three} N ₇ O _Five Molecular weight 417.43) Example 24. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-6-[(methoxycarbonyl
Ru) azo] -5-methyl-azirino [2 ', 3': 3,
4] Preparation of pyrrolo [1,2-a] indole Mitomycin A 300 mg was dissolved in methanol 35 ml.
Then, add methoxycarbonylhydra to this solution with stirring.
Add 200 mg of gin and stir at room temperature for 24 hours. Under reduced pressure
The solvent was distilled off and the residue was subjected to silica gel column chromatography.
Chloroform-methanol (9: 1; v /
Separated and purified with v) as the developing solvent, 140 mg of the title compound
Is obtained as a reddish orange powder (yield 40.0%). ¹ H-NMR (d ₆ -DMSO) δ: 2.01 (3H, s), 2.75 (1H, m),
2.84 (1H, m), 3.14 (3H, s), 3.39 (1H, dd), 3.49 (1H, d), 3.77
(3H, s), 4.06 (1H, dd), 4.09 (1H, d), 4.64 (1H, dd), 6.52 (2H, d)
br), 9.48 (1H, br), 15.61 (1H, s) IR (KBr) cm ^-1 3400, 1708, 1620, 1490, 1455, 1392, 1338, 124
0, 1176, 1065 MASS 408 (C ₁₇ H _{twenty one} N _Five O ₇ Molecular weight 407.38) Example 25. [1aS- (1aα, 8β, 8aα, 8bα)]-6-
[(Aminocarbonyl) azo] -8-{[aminocarbo
Nyl) oxy] methyl} -4,7-dihydroxy-1,
1a, 2,8,8a, 8b-hexahydro-8a-meth
Xy-5-methyl-azirino [2 ', 3': 3,4] pi
Production of Lolo [1,2-a] indole By the same method as in Example 24, mitomycin A30
0 mg, semicarbazide hydrochloride 190 mg and triethylamid
200 mg of the title compound (yield 59.
3%). ¹ H-NMR (d ₆ -DMSO) δ: 2.04 (3H, s), 2.75 (1H, m),
2.81 (1H, m), 3.14 (3H, s), 3.42 (1H, d), 3.42 (1H, dd), 4.07
(1H, d), 4.08 (1H, dd), 4.67 (1H, dd), 6.14 (2H, br), 6.95 (2
H, s), 8.83 (1H, s), 14.84 (1H, s) IR (KBr) cm ^-1 3450, 3196, 1705, 1616, 1556, 1487, 1411, 134
0, 1199, 1170, 1061, 935 MASS 393 (C ₁₆ H ₂₀ N ₆ O ₆ Molecular weight 392.37) Example 26. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-5-methyl-6-{[(phen
Nyl) aminocarbonyl] azo} -azirino [2 ′,
Production of 3 ': 3,4] pyrrolo [1,2-a] indole Dissolve 300 mg of mitomycin A in methanol and stir.
While adding 4-phenylsemicarbazide 210 to this solution,
Add mg and stir at room temperature for 7 hours. The crystals that have precipitated
Separate and wash the crystals several times with ether
This gave 190 mg of the title compound as red-orange crystals.
(Yield 47.6%). ¹ H-NMR (d ₆ -DMSO) δ: 2.12 (3H, s), 2.76 (1H, m),
2.86 (1H, d), 3.16 (3H, s), 3.41 (1H, dd), 3.49 (1H, d), 3.50
(1H, dd), 4.09 (1H, d), 4.76 (1H, dd), 6.51 (2H, br), 7.04 (1
H, m), 7.32 (1H, m), 7.61 (1H, m), 9.64 (1H, s), 15.53 (1H, s) IR (KBr) cm ^-1 3450, 3308, 1705, 1604, 1539, 1481, 1446, 132
9, 1153, 1059, 933 MASS 469 (C _{twenty two} H _{twenty four} N _Four O ₆ Molecular weight 468.47) Example 27. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Benzoyl) azo] -3,8a-dimethoxy-1,
1a, 2,8,8a, 8b-hexahydro-7-hydro
Xy-5-methyl-azirino [2 ', 3': 3,4] pi
Preparation of Lolo [1,2-a] indole 1.09 g of the compound of Example 17 was dissolved in 100 ml of DMF.
Then, add 0.14 g of sodium methoxide at room temperature for 3
Stir for 0 minutes, add 0.22 ml of methyl iodide and add to room
Stir at temperature for 2.5 hours. Evaporate the solvent under reduced pressure
Chloroform and water were added to the obtained oily substance and extracted.
The chloroform layer was dried over anhydrous sodium sulfate, and the pressure was reduced.
The chloroform is distilled off underneath. Silica gel color residue
Chromatography-chloroform-acetone
Separated and purified using (1: 1; v / v) as a developing solvent. this
The resulting oil is further chromatographed on thin layer silica gel.
Chloroform-acetone (1: 2; v /
Purification using v) as a developing solvent yields 140 mg of the title compound.
0.14 g is obtained as a tan powder (yield 13.1).
%). ¹ H-NMR (CDCl _Three ) Δ: 2.28 (3H, s), 2.88 (1H, m), 2.99
(1H, d), 3.26 (3H, s), 3.57 (1H, d), 3.73 (3H, s), 3.75 (1H, d
d), 4.22 (1H, d), 4.57 (1H, dd), 4.84 (2H, br), 4.92 (1H, dd),
7.55 (3H, m), 8.05 (2H, m), 17.08 (1H, s) IR (KBr) cm ^-1 3430, 1696, 1608, 1573, 1480, 1452, 1380, 132
0, 1239, 1065, 982 MASS 468 (C _{twenty three} H _{twenty five} N _Five O ₆ Molecular weight 467.49) Example 28. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-6-[(2 "-hydroxybenzoyl) azo]
-8a-methoxy-5-methyl-azirino [2 ',
Production of 3 ′: 3,4] pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A21 was prepared.
5 mg, salicyl hydrazide 90 mg and potassium carbonate 10
148 mg of the title compound is obtained from 0 mg (yield 53.6).
%). ¹ H-NMR (CDCl _Three ) Δ: 2.08 (3H, s), 2.77 (1H, d), 2.86
(1H, d), 3.16 (3H, s), 3.50 (1H, d), 4.10 (1H, d), 4.10 (1H, d
d), 4.71 (1H, dd), 6.54 (2H, br), 6.97 (2H, m), 7.44 (1H, m),
7.85 (1H, m), 16.52 (1H, s) IR (KBr) cm ^-1 3400, 1710, 1648, 1485, 1405, 1340, 1240, 116
0, 1072, 928 MASS 470 (C _{twenty two} H _{twenty three} N _Five O ₇ Molecular weight 469.45) Example 29. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Benzoyl) azo] -1,5-dimethyl-1,1
a, 2,8,8a, 8b-hexahydro-4,7,8a
-Trihydroxy-azirino [2 ', 3': 3,4] pi
Production of Rolo [1,2-a] indole By the same method as in Example 17, mitomycin B1.
25 g, benzoylhydrazine 1.21 g and potassium carbonate
0.90 g of the title compound is obtained from 0.30 g of ammonium (yield
69.6%). ¹ H-NMR (d ₆ -DMSO) δ: 2.06 (3H, s), 2.21 (3H, s),
2.29 (1H, d), 2.47 (1H, d), 3.50 (1H, dd), 3.57 (1H, d), 4.04
(1H, d), 4.14 (1H, dd), 4.78 (1H, dd), 6.42 (2H, br), 6.44 (1
H, s), 7.62 (3H, m), 7.90 (2H, dd), 9.80 (1H, s), 17.23 (1H, s) IR (KBr) cm ^-1 3440, 3198, 1704, 1662, 1616, 1576, 1488, 145
5, 1354, 1239, 1167, 1068 MASS 454 (C _{twenty two} H _{twenty three} N _Five O ₆ Molecular weight 453.6) Example 30. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-6-[(tert-butoxycarbonyl) azo]-
8a-Methoxy-5-methyl-azirino [2 ', 3':
Production of 3,4] pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A25 was prepared.
0 mg, tert-butoxycarbonylhydrazine 284 mg and
138 mg of the title compound is obtained from 150 mg of potassium carbonate.
(Yield 42.7%). ¹ H-NMR (d ₆ -DMSO) δ: 1.48 (9H, s), 2.00 (3H, s),
2.75 (1H, m), 2.84 (1H, m), 3.14 (3H, s), 3.35 (1H, dd), 3.48
(1H, d), 4.07 (1H, d), 4.08 (1H, dd), 4.65 (1H, dd), 6.50 (2H, d
br), 9.36 (1H, br), 15.41 (1H, s) IR (KBr) cm ^-1 3446, 3312, 1723, 1698, 1615, 1557, 1532, 148
1, 1375, 1213, 1162, 1056, 1033, 934 MASS 450 (C ₂₀ H ₂₇ N _Five O ₇ Molecular weight 449.46) Example 31. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -4,7-
Dihydroxy-1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-5-methyl-6-{[(meth
Ruthio) thiocarbonyl] azo} -azirino [2 ′,
Production of 3 ′: 3,4] pyrrolo [1,2-a] indole By the same method as in Example 17, mitomycin A30 was prepared.
0 mg [(methylthio) thiocarbonyl] hydrazine 32
0 mg and potassium carbonate 145 mg to 267 mg of the title compound
Is obtained (yield 70.7%). ¹ H-NMR (d ₆ -DMSO) δ: 2.04 (3H, s), 2.57 (3H, s),
2.77 (1H, m), 2.84 (1H, m), 3.16 (3H, s), 3.40 (1H, dd), 3.53
(1H, d), 4.09 (1H, dd), 4.11 (1H, d), 4.64 (1H, dd), 6.54 (2H, d
br), 10.00 (1H, s), 16.95 (1H, s) IR (KBr) cm ^-1 3280, 2930, 1707, 1610, 1578, 1485, 1395, 131
5, 1250, 1134, 1065, 932 MASS 440 (C ₁₇ H _{twenty one} N _Five O _Five S molecular weight 439.51) Example 32. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Aminothiocarbonyl) azo] -4,7-dihydro
Xy-1,1a, 2,8,8a, 8b-hexahydro-
8a-Methoxy-5-methyl-azirino [2 ', 3':
Production of 3,4] pyrrolo [1,2-a] indole By the same method as in Example 24, mitomycin A0.
30 g and thiosemicarbazide 0.39 g
80 mg of the product is obtained (yield 22.8%). ¹ H-NMR (d ₆ -DMSO) δ: 2.05 (3H, s), 2.76 (1H, m),
2.86 (1H, m), 3.16 (3H, s), 3.41 (1H, dd), 3.48 (1H, d), 4.09
(1H, d), 4.10 (1H, dd), 4.67 (1H, dd), 6.46 (2H, br), 8.93 (1
H, s), 15.60 (1H, s) IR (KBr) cm ^-1 3410, 1707, 1600, 1570, 1453, 1379, 1213, 115
3, 1060, 925 MASS 409 (C ₁₆ H ₂₀ N ₆ O ₃ S molecular weight 408.44) Example 33. [1aS- (1aα, 8β, 8aα, 8bα)]-8-
{[(Aminocarbonyl) oxy] methyl} -6-
[(Benzoyl) azo] -7,8a-dihydroxy-
1,5-dimethyl-1,1a, 2,8,8a, 8b-f
Xahydro-4-methoxy-azirino [2 ', 3':
Preparation of 3,4] pyrrolo [1,2-a] indole Using a method similar to that in Example 27, the compound of Example 29 was used.
553 mg, sodium methoxide 72 mg and iodide
80 mg of the title compound was obtained from 0.15 ml of chill (yield 1
4.0%). ¹ H-NMR (d ₆ -DMSO) δ: 2.11 (3H, s), 2.21 (3H, s),
2.32 (1H, m), 3.51 (1H, dd), 3.61 (1H, d), 3.69 (3H, s), 3.90
(1H, d), 4.16 (1H, dd), 4.78 (1H, dd), 6.53 (1H, s), 6.45 (2H, d
br), 7.66 (3H, m), 7.92 (2H, m), 17.38 (1H, s) IR (KBr) cm ^-1 3410, 1729, 1691, 1616, 1506, 1487, 1452, 138
7, 1322, 1232, 1170, 1061, 960 MASS 468 (C _{twenty three} H _{twenty five} N _Five O ₆ Molecular weight 467.49) Example 34. [1aS- (1aα, 8β, 8aα, 8bα)]-4-
Acetyloxy-8-{[(aminocarbonyl) oxy
Ci] methyl} -1,1a, 2,8,8a, 8b-hexa
Hydro-8a-methoxy-5-methyl-azirino
[2 ', 3': 3,4] pyrrolo [1,2-a] indole
Manufacture of 0.66 g of the compound obtained in Example 11 in methanol 3
Dissolve in 0 ml, 0.3% 15% hydroxylamine aqueous solution
Add 1 ml and stir at room temperature for 1 hour. 15% more hydro
Add 0.31 ml of aqueous xylamine solution and stir for 40 minutes
It The solvent was distilled off under reduced pressure, and the resulting oily substance was washed with chloroform.
-Methanol (15: 1; v / v) was used as the developing solvent.
After separation and purification, 0.08 g of the title compound was obtained (yield 14.3).
%). ¹ H-NMR (CDCl _Three ) Δ: 2.27 (3H, s), 2.37 (3H, s), 2.86
(1H, dd), 2.98 (1H, d), 3.18 (3H, s), 3.43 (1H, dd), 3.70 (1H,
dd), 3.92 (1H, d), 4.50 (1H, dd), 4.80 (1H, dd), 4.95 (2H, b
r), 18.66 (1H, s) IR (KBr) cm ^-1 3400, 2950, 1710, 1620, 1480, 1340, 1210, 119
0, 1070 MASS 393 (C ₁₇ H ₂₀ N _Four O ₇ Molecular weight 392.38) Reference Example 1 Injectable formulation example 10 mg of the compound obtained in Example 1 was used in 10 ml of a sterile brown bag.
Dispense into the ear to make a sterile powder formulation. Sterilize this before use 5
Add 5 ml of 0% ethanol water, shake and stir well to dissolve,
Prepare an injection solution.

Reference Example 2 Tablet formulation example 20 mg of the compound of Example 6, 170 mg of lactose, 20 mg of potato starch, 4 mg of hydroxypropyl cellulose,
A tablet is prepared by a conventional method with a compounding ratio of 1 mg of magnesium stearate.

Reference Example 3 Suppository formulation example 20 mg of the compound of Example 8 and Witepsol H-15 75
A suppository is prepared by a conventional method at a compounding ratio of 0 mg and Witepsol E-75 320 mg.

EFFECTS OF THE INVENTION Compound (I) has excellent antitumor activity and can be used as an active ingredient of an antitumor agent or as an intermediate for producing a compound having a further useful antitumor activity.

 ─────────────────────────────────────────────────── ─── Continuation of the front page Examiner Norihiro Sano

Claims (1)

[Claims] 1. A formula {In the formula, A is ON- or R ₄ N = N- [wherein, R ₄ is Or an unsubstituted or substituted heterocyclic group (in these formulas, R ₅ is a hydrogen atom, lower alkyl or lower cycloalkyl, R ₆ is a hydrogen atom, a halogen atom, hydroxy,
Lower alkoxy, amino or nitro, R ₇ is lower alkyl or lower cycloalkyl, X is an oxygen atom, sulfur atom or imino, R ₈ is the same or different from R ₅ and is a hydrogen atom, lower alkyl or lower Is a cycloalkyl)], and R ₁ and R ₂ are the same or different and each is a hydrogen atom, lower alkyl, lower cycloalkyl, unsubstituted or substituted aralkyl, unsubstituted or substituted lower alkanoyl, unsubstituted or substituted arylcarbonyl, non-substituted. A substituted or substituted lower alkanesulfonyl, arylsulfonyl or aralkylsulfonyl, R ₃ is a hydrogen atom or carbamoyl, Y is a hydrogen atom or methyl, Z is a hydrogen atom, methyl or acetyl, Represents an α-bond or a β-bond}.